hnRNPA2B1 agonist compounds and their anti-infective applications

Abstract

This disclosure relates to heterogeneous nuclear ribonucleic acid A2B1 (hnRNPA2B1) agonist compounds and their anti-infective applications. Specifically, this disclosure relates to fused ring compounds of formula (I), their cis-trans isomers, their enantiomers, their diastereomers, their racemic mixtures, their solvates, their hydrates, or their pharmaceutically acceptable salts or prodrugs thereof, compositions and products comprising any of the former, and the use of the aforementioned substances as hnRNPA2B1 agonists in the preparation of products for the prevention and / or treatment of infectious diseases and / or infection-related diseases and / or symptoms. This disclosure has broad application prospects for the broad-spectrum prevention and treatment of infections (such as viral infections).

Description

Technical Field

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

[0002] Infections, especially viral infections, are a common and highly dangerous clinical disease. Initially, the molecular mechanisms by which the body fights viral infections were not fully understood. However, with the discovery of interferon, a class of cytokines, the molecular biological basis of the innate and acquired immune cells produced by the body in response to viral infections and their functions has gradually been understood.

[0003] Interferon (IFN) is a family of cytokines with potent antiviral functions. In 1957, Professors Alick Isaacs and Jean Lindenmann discovered a component during their research on influenza virus infection in chicken embryos. This component significantly inhibited the proliferation of the influenza virus, and they named it interferon (Isaacs, A. et al., Proc R Soc Lond B Biol Sci. 1957; 927:258-267.). Subsequently, cytokines in the interferon family were found to possess broad-spectrum and effective antiviral effects. Currently, interferon is widely used clinically to prevent and treat various diseases caused by viral infections.

[0004] Type I interferon (IFN-I) plays a crucial role in the host's defense against viral, bacterial, parasitic, and fungal infections by directly or indirectly activating and regulating innate and adaptive immune cells through the induction of other mediators. It is a core cytokine in the host's defense against pathogens such as viruses. In infected and neighboring cells, type I interferon induces the expression of interferon-stimulated genes (ISGs), inhibiting further pathogen transmission. Innate immune cells also respond to type I interferon by enhancing antigen presentation and increasing the production of cytokines and chemokines. Adaptive immunity is also influenced by type I interferon: for example, it can induce B cells to produce specific antibodies and amplify the effector functions of T cells.

[0005] Innate immune receptors recognize pathogen-associated molecular patterns (PAMPs) or damage-associated molecular patterns (DAMPs), inducing the production of cytokines such as type I interferon and inflammatory factors, thus initiating an innate immune response. Among these PAMPs and DAMPs, pathogen-derived and autologous nucleic acids are crucial ligands. Therefore, agonists targeting these innate immune recognition receptors have become an important means of treating infectious diseases.

[0006] Nucleic acid innate immune recognition receptors include RNA recognition receptors and DNA recognition receptors. Based on their intracellular location, DNA recognition receptors are mainly divided into cytoplasmic and nuclear DNA recognition receptors. The main DNA recognition receptor in the cytoplasm is cyclic GMP-AMP synthase (cGAS). After recognizing DNA in the cytoplasm, it induces the activation of the Stimulator of Interferon Genes (STING), which further recruits and activates TANK-binding kinase 1 (TBK1) and interferon-regulatory factor 3 (IRF3), initiating the production of type I interferon. In addition, TLR9 recognizes single-stranded DNA containing unmethylated CpG motifs, and AIM2 (Absent in melanoma-2) can also recognize pathogen-derived DNA in the cytoplasm, promoting the formation of inflammasomes and the maturation and release of the inflammatory cytokine IL-1β.

[0007] Existing studies have identified many proteins that can recognize viral DNA and induce the production of IFN-α / β, such as RNA polymerase III, IFI16, DAI, LRRFIP1, LSm14A, MRE11, DNA-PK, HMGBs, DDX41, and cyclic GMP-AMP (cGAMP) synthase (cGAS) (Goubau, D. et al.; Immunity, 2013; 38, 855-869.). After recognizing the virus, these PPRs stimulate immune signaling pathways within the host, inducing the production of large amounts of type I interferon, thereby rapidly establishing the body's first line of defense against infection. However, only cytoplasmic cGAS and DNA-PK have been confirmed as DNA recognition receptors in mouse in vivo experiments. Several other proteins are also involved in DNA virus-induced inflammatory responses, including AIM2, IFI16, Rad50, and Sox2. Therefore, in-depth research on nuclear DNA recognition is urgently needed to fully and clearly understand the innate immune response against DNA viruses, especially to find the mechanism that links the recognition of exogenous and intrigued DNA with the activation of extranuclear innate immune signaling.

[0008] Heterogeneous nuclear ribonucleoprotein A2B1 (hnRNP-A2B1) is a newly identified DNA recognition receptor in the cell nucleus, belonging to the hnRNP family. On one hand, hnRNP-A2B1 can sense and recognize the nucleic acid components of DNA viruses (such as HSV-1), and self-activate to form a homodimer. Demethylation is then mediated by the demethylase JMJD6, causing the receptor to translocate from the nucleus to the cytoplasm, thereby activating the TBK1-IRF3 signaling pathway and initiating type I interferon expression to exert an antiviral effect. On the other hand, as an RNA-binding protein, hnRNPA2B1 can also promote m6A modification, nucleoplasmic translocation, and translation on cGAS, IFI16, and STING mRNA, thus ensuring sufficient induction of type I interferon expression to resist viral infection.

[0009] In summary, the development of specific agonists targeting hnRNPA2B1 holds great potential for the treatment of infectious diseases. There is an urgent need in this field to develop an immunologically active substance that can recognize viral DNA in the cell nucleus, initiate interferon production, enhance antiviral effects, effectively resist viral infection, and control damage caused by viral infection. Summary of the Invention

[0010] This disclosure provides fused cyclic compounds having the structure shown in formula (I), related derivatives thereof (e.g., cis-trans isomers, enantiomers, diastereomers, racemic mixtures, solvates, hydrates, or pharmaceutically acceptable salts thereof, or prodrugs thereof), and products comprising said compounds or related derivatives thereof. This disclosure also provides the use of said compounds, derivatives, and products in anti-infective applications, and further provides their use in treating or preventing infectious diseases and related diseases or symptoms. The compounds, pharmaceuticals, pharmaceutical compositions, or kits of this disclosure can be used to effectively combat infections and control the occurrence of infectious diseases.

[0011] In some aspects of this disclosure, fused ring compounds of formula (I), their cis-trans isomers, their enantiomers, their diastereomers, their racemic mixtures, their solvates, their hydrates, or pharmaceutically acceptable salts thereof, or their prodrugs, are provided.

[0012]

[0013] Where L is -(CH2)n-, and n is an integer from 0 to 6;

[0014] X is a halogen;

[0015] R 1 For -NR 4 R 5 , where R 4 R 5 Each independently is H or C 1-6 alkyl;

[0016] R 2 and R 3 Each independently is H or C 1-6 alkyl;

[0017] Or R 2 and R 3 Together with the N atoms to which they are attached, they form 5-8 membered heterocyclic alkyl groups containing 1-3 heteroatoms selected from nitrogen, oxygen, or sulfur;

[0018] The C 1-6 The alkyl group and the 5-8 membered heterocyclic alkyl group are either unsubstituted or substituted by one or more substituents selected from the group consisting of: hydroxyl, amino, halogen, C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Alkoxy, hydroxy C 1-6 Alkyl, C 1-6 Alkylamino, C 3-6 cycloalkyl, C 3-6 cycloalkyl C 1-6Alkyl, 3-6 membered heterocyclic alkyl, 3-6 membered heterocyclic alkyl C 1-6 Alkyl, 6-10 aryl, 6-10 aryl C 1-6 Alkyl or 5-10 heteroaryl and 5-10 heteroaryl C 1-6 alkyl.

[0019] Among some aspects of this disclosure, the use of the compounds of the present invention in the preparation of products for the prevention and / or treatment of infectious diseases and / or infection-related diseases and / or symptoms is provided.

[0020] In some aspects of this disclosure, a method for preventing and / or treating infectious diseases and / or infection-related diseases and / or symptoms is also provided, the method comprising administering to a subject in need a preventive and / or therapeutically effective amount of a compound or product of this disclosure.

[0021] In some aspects of this disclosure, compounds or products of this disclosure are also provided for the prevention and / or treatment of infectious diseases and / or diseases and / or symptoms related to infection.

[0022] Among some aspects of this disclosure, the use of the compounds or products of this disclosure in increasing the levels of interferons (such as type I interferons, for example IFN-α and / or IFN-β).

[0023] Those skilled in the art can combine the foregoing technical solutions and features in any way without departing from the inventive concept and protection scope of this disclosure. Other aspects of this disclosure will be apparent to those skilled in the art due to the content of this disclosure. Attached Figure Description

[0024] The present disclosure will be further described below with reference to the accompanying drawings, which are shown only for illustrating the embodiments of the present disclosure and are not intended to limit the scope of the present disclosure.

[0025] Figure 1 The purpose of this study was to test the activation effect of the compound on the nucleus exit of macrophage hnRNPA2B1 cells.

[0026] Figure 2 The purpose of this study was to test the activation effect of the compound on downstream TBK1 and IRF3 cells of macrophage hnRNPA2B1 cells after they exit the nucleus.

[0027] Figures 3-5 The purpose of this study was to test the activation effect of the compound on type I interferon in macrophages.

[0028] Figures 6-7 The test compound was used to activate interferon-stimulated genes in immune cells.

[0029] Figure 8The test compound was used to inhibit the replication of herpes simplex virus HSV-1.

[0030] Figure 9 The purpose of this study was to test the inhibitory effect of the compound on the replication of hepatitis B virus (HBV).

[0031] Figure 10 The purpose of this study was to test the inhibitory effect of the compound on the replication of vesicular stomatitis virus (VSV). Detailed Implementation

[0032] Through extensive research, development, and experimentation, this application has discovered a novel class of fused-ring compounds that can bind specifically to the hnRNPA2B1 protein with high affinity and can further induce higher levels of IFN-β, and is therefore defined as an hnRNPA2B1 agonist.

[0033] In anti-cellular infection assays against different types of infectious agents, the hnRNPA2B1 agonist compounds of this disclosure significantly inhibited the replication of infectious agents, thereby exerting an anti-infective effect. Furthermore, since the hnRNPA2B1 agonist compounds of this disclosure can broadly induce higher levels of IFN-β, it is expected that these compounds will exert anti-infective effects through type I interferon action.

[0034] Therefore, this disclosure provides methods and strategies for using agonist compounds of the novel anti-infective molecule hnRNPA2B1 to inhibit infection, or for the prevention and treatment of infectious diseases, particularly for controlling viral infections, such as liver damage caused by viral infections.

[0035] All numerical ranges provided herein are intended to clearly include all values ​​falling between the endpoints of the range and the range of values ​​between them. Features mentioned in this disclosure or in the embodiments may be combined. All features disclosed in this specification may be used in any compositional form, and each feature disclosed in the specification may be replaced by any alternative feature that provides the same, equivalent, or similar purpose. Therefore, unless otherwise specified, the disclosed features are merely general examples of equivalent or similar features.

[0036] As used in this article, “containing,” “having,” or “including” includes “containing,” “mainly composed of,” “substantially composed of,” and “composed of”; “mainly composed of,” “substantially composed of,” and “composed of” are subordinate concepts of “containing,” “having,” or “including.”

[0037] hnRNPA2B1 agonist compound

[0038] As used herein, the terms "hnRNPA2B1 protein (peptide)" and "hnRNPA2B1" are used interchangeably and refer to the heterogeneous nuclear ribonucleoprotein A2B1. The hnRNPA2B1 protein involved in this disclosure may be a protein encoded by the hnRNPA2B1 gene, its cDNA, or CDS in an animal (e.g., human or mouse), or a homologous sequence of such protein that promotes interferon expression (e.g., homologous sequences of hnRNPA2B1 can be obtained from databases or alignment software known in the art), variant, or modified form.

[0039] As used herein, the terms “hnRNPA2B1 gene,” “hnRNPA2B1 encoding gene,” “hnRNPA2B1 protein encoding gene,” or “nucleic acid molecule encoding hnRNPA2B1” are used interchangeably and refer to a nucleotide sequence encoding the hnRNPA2B1 protein or polypeptide described herein, which may be, for example, the human hnRNPA2B1 gene with Gene ID: 3181, or the mouse hnRNPA2B1 gene with Gene ID: 53379. The term also includes molecules that hybridize with a labeled nucleic acid molecule under stringent conditions, or family gene molecules highly homologous to the aforementioned molecules, whose expression is believed to promote interferon production and its effects.

[0040] This disclosure provides a novel class of compounds that act as hnRNPA2B1 "agonists" (or "promoters"). The terms "agonist" or "hnRNPA2B1 agonist compound" are used interchangeably to refer to a class of novel fused-ring compounds that can increase the level or activity of hnRNPA2B1, specifically bind to the hnRNPA2B1 protein with high affinity, and further broadly induce higher levels of IFN-β, thereby exerting an agonistic effect on hnRNPA2B1.

[0041] The hnRNPA2B1 agonist compound disclosed herein can inhibit infection, and thus can be further used for the prevention or treatment of diseases associated with infection (especially viral infection), and / or symptoms caused by infection, as well as chronic inflammatory diseases caused by infection, and / or their symptoms.

[0042] A fused ring compound of formula (I), its cis-trans isomer, its enantiomer, its diastereomer, its racemic mixture, its solvate, its hydrate, or a pharmaceutically acceptable salt thereof, or a prodrug thereof.

[0043]

[0044] Where L is -(CH2)n-, and n is an integer from 0 to 6;

[0045] X is a halogen;

[0046] R 1 For -NR 4 R 5 , where R 4 R 5 Each independently is H or C 1-6 alkyl;

[0047] R 2 and R 3 Each independently is H or C 1-6 alkyl;

[0048] Or R 2 and R 3 Together with the N atoms to which they are attached, they form 5-8 membered heterocyclic alkyl groups containing 1-3 heteroatoms selected from nitrogen, oxygen, or sulfur;

[0049] The C 1-6 The alkyl group and the 5-8 membered heterocyclic alkyl group are either unsubstituted or substituted by one or more substituents selected from the group consisting of: hydroxyl, amino, halogen, C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Alkoxy, hydroxy C 1-6 Alkyl, C 1-6 Alkylamino, C 3-6 cycloalkyl, C 3-6 cycloalkyl C 1-6 Alkyl, 3-6 membered heterocyclic alkyl, 3-6 membered heterocyclic alkyl C 1-6 Alkyl, 6-10 aryl, 6-10 aryl C 1-6 Alkyl or 5-10 heteroaryl and 5-10 heteroaryl C 1-6 alkyl.

[0050] Preferably, X is F or Cl, more preferably, X is F.

[0051] Preferably, L is -(CH2)n-, where n is 1, 2 or 3, more preferably, n is 2.

[0052] Preferably, R 4 R 5 Each can be H, methyl, or ethyl.

[0053] Preferably, R 2 and R 3 Each independently is C 1-3 Alkyl, preferably methyl, ethyl or isopropyl, wherein the C 1-3 The alkyl group is unsubstituted or substituted with a 3-8 membered heterocyclic alkyl group containing 1-2 heteroatoms selected from nitrogen, oxygen, or sulfur; more preferably, R 2 and R 3Each independently is C 1-3 Alkyl, preferably methyl, ethyl or isopropyl, wherein the C 1-3 The alkyl group is either unsubstituted or substituted with a 4-6 membered heterocyclic alkyl group containing one nitrogen atom; most preferably, R 2 and R 3 Each independently is C 1-3 The alkyl group, preferably methyl, ethyl or isopropyl, wherein the C1-3 alkyl group is unsubstituted or substituted with pyrrolidinyl group.

[0054] Preferably, R 2 and R 3 Together with the N atoms they are attached to, they form a 5-8 membered heterocyclic alkyl group containing two nitrogen atoms, wherein the 5-8 membered heterocyclic alkyl group is bonded to a 3-8 membered heterocyclic alkyl group C. 1-6 Alkyl or hydroxy C 1-6 Alkyl substitution; more preferably, R 2 and R 3 Together with the N atoms they are attached to, they form a 6-membered heterocyclic alkyl group containing 2 nitrogen atoms, wherein the 6-membered heterocyclic alkyl group is bonded to a 4-6-membered heterocyclic alkyl group C. 1-6 Alkyl or hydroxy C 1-6 Alkyl substitution; more preferably, R 2 and R 3 Together with the N atoms to which they are attached, they form a piperazine group, which is bonded by a 4-6 membered heterocyclic alkyl group. 1-6 Alkyl or hydroxy C 1-6 Alkyl substitution; most preferably, R 2 and R 3 Together with the N atom to which they are attached, they form a piperazine group, which is bonded to a 5-6 membered heterocyclic alkyl group containing one N atom. 1-6 Alkyl substitution or hydroxyl C 1-6 Alkyl substitution, preferably, the piperazine group is replaced by a pyrrolidinyl C 1-6 Alkyl substitution or hydroxyl C 1-6 Alkyl substitution, preferably, the piperazine group is substituted with pyrrolyl ethyl or hydroxyethyl.

[0055] Preferably, the compound has the following formula (II):

[0056]

[0057] Among them, R a 、 R b Each independently is H or C 1-6 Alkyl, preferably, R a 、 R b Each can be independently H, methyl, or ethyl;

[0058] M is -(CH2)n-, where n is 1, 2 or 3, more preferably n is 2;

[0059] X is F or Cl, more preferably, X is F;

[0060] R c It is a hydroxyl, amino, or 3-8 membered heterocyclic alkyl group containing 1-2 heteroatoms selected from nitrogen, oxygen, or sulfur, more preferably, R c It is a hydroxyl group or a 4-6 membered heterocyclic alkyl group containing one nitrogen atom, more preferably, R c It is hydroxyl or pyrrolidinyl, most preferably, R c It is hydroxyl or 1-pyrrolidinyl.

[0061] Preferably, the compound is selected from the group consisting of compounds or salts thereof (e.g., hydrochlorides):

[0062]

[0063] Unless otherwise stated, the following terms and phrases as used herein are intended to have the following definitions. A particular term or phrase should not be considered uncertain or unclear unless specifically defined, but should be understood in its ordinary sense. When a trade name appears in this document, it is intended to refer to the corresponding product or active ingredient.

[0064] The term “pharmaceutically acceptable” as used herein refers to compounds, materials, compositions, and / or dosage forms that, within the bounds of reliable medical judgment, are suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic reactions, or other problems or complications, in proportion to a reasonable benefit / risk ratio.

[0065] The term "pharmaceutically acceptable salt" refers to a salt of the compounds of this invention, prepared by reacting a compound having specific substituents discovered in this invention with a relatively non-toxic acid or base. When the compounds of this invention contain relatively acidic functional groups, a base addition salt can be obtained by contacting the neutral form of such compounds with a sufficient amount of base in a pure solution or a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amine, or magnesium salts or similar salts. When the compounds of this invention contain relatively basic functional groups, a base addition salt can be obtained by contacting the neutral form of such compounds with a sufficient amount of acid in a pure solution or a suitable inert solvent. Pharmaceutically acceptable examples of acid addition salts include inorganic acid salts, such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, hydrogen sulfate, hydroiodic acid, phosphorous acid, etc., and organic acid salts, such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, octanoic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, and methanesulfonic acid; salts of amino acids (such as arginine); and salts of organic acids such as glucuronic acid. Certain specific compounds of the present invention contain both basic and acidic functional groups, and thus can be converted into either a base or an acid addition salt.

[0066] The pharmaceutically acceptable salts of the present invention can be synthesized from parent compounds containing acid radicals or bases by conventional chemical methods. Generally, such salts are prepared by reacting these compounds in free acid or base form with a stoichiometric amount of a suitable base or acid in water or an organic solvent or a mixture thereof.

[0067] The compounds of this invention can exist in specific geometric or stereoisomeric forms. This invention contemplates all such compounds, including cis and trans isomers, (-)- and (+)- enantiomers, (R)- and (S)- enantiomers, diastereomers, (D)- isomers, (L)- isomers, and racemic mixtures thereof, as well as other mixtures, such as mixtures enriched with enantiomers or diastereomers, all of which are within the scope of this invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers and mixtures thereof are included within the scope of this invention.

[0068] Unless otherwise stated, the terms "enantiomer" or "optical isomer" refer to stereoisomers that are mirror images of each other.

[0069] Unless otherwise stated, the terms "cis-trans isomers" or "geometric isomers" arise because the single bonds of double bonds or cyclic carbon atoms cannot rotate freely.

[0070] Unless otherwise stated, the term "diastereomer" refers to a stereoisomer of a molecule having two or more chiral centers and being a non-mirror image of each other.

[0071] The compounds of this invention can exist in specific forms. Unless otherwise stated, the terms "tautomer" or "tautomer form" refer to isomers of different functional groups in dynamic equilibrium at room temperature, capable of rapidly interconverting into each other. If tautomerization is possible (e.g., in solution), chemical equilibrium of the tautomer may be achieved. For example, proton tautomers (also called prototropic tautomers) include interconversions via proton migration, such as keto-enol isomerization and imine-enamine isomerization. Valence tautomers include interconversions via the recombination of some bonding moieties. A specific example of keto-enol tautomerization is the interconversion between the two tautomers, pentane-2,4-dione and 4-hydroxy-3-en-2-one.

[0072] The term "prodrug" generally refers to a compound of general formula (I) that has undergone functional group derivatization, and whose derivative can be readily converted into a compound of general formula (I) in vivo. Suitable prodrug selection and preparation methods are typically described in, for example, Design of Prodrugs, ed. H. Bundgaard, Elsevier, 1985.

[0073] The compounds of the present invention may contain, on one or more atoms constituting the compound, atomic isotopes in non-natural proportions, said isotopes having the same number of atoms but different atomic masses or mass numbers from those that are predominantly found in nature. For example, compounds may be labeled with radioactive isotopes, such as deuterium (…). 2 H), tritium ( 3 H), Iodine-125 ( 125 I) or C-14 14C). All isotopic variations of the compounds of this invention, regardless of radioactivity, are included within the scope of this invention. Isotopic variants may enhance certain therapeutic advantages, such as the use of deuterium to replace hydrogen to form deuterated drugs. The bond between deuterium and carbon is stronger than that between ordinary hydrogen and carbon. Compared to undeuterated drugs, deuterated drugs have advantages such as reduced toxicity, increased drug stability, enhanced efficacy, and prolonged drug biological half-life. Alternatively, they may provide standard compounds that can be used for characterization of biological samples. Isotope-enriched compounds of general formula (I) can be prepared without extensive experimentation using conventional techniques well known to those skilled in the art, or by methods similar to those described in the routes and embodiments of this invention, using appropriate isotope-enriched reagents and / or intermediates.

[0074] "Optional" or "optionally" means that the event or condition described below may occur but is not required to occur, and the description includes both the scenario in which said event or condition occurs and the scenario in which said event or condition does not occur.

[0075] The naming conventions used in this invention are based on the IUPAC system naming generated by ChemDraw software. Any open valence bonds appearing on carbon, oxygen, sulfur, or nitrogen atoms in the structures given in this invention indicate the presence of hydrogen atoms.

[0076] The term "substituted" means that any one or more hydrogen atoms on a specific atom are replaced by a substituent, which can include deuterium and hydrogen variants, provided that the valence state of the specific atom is normal and the substituted compound is stable. When the substituent is oxo (i.e., =O), it means that two hydrogen atoms are replaced; oxygen substitution does not occur on the aromatic group. The term "optionally substituted" means that it may or may not be substituted, unless otherwise specified; the type and number of substituents can be arbitrary on a chemically feasible basis.

[0077] When any variable (e.g., R) appears more than once in the composition or structure of a compound, its definition is independent in each case. Thus, for example, if a group is substituted by 0-2 Rs, the group can optionally be substituted by at most two Rs, and the Rs in each case have independent options. Furthermore, combinations of substituents and / or their variants are only permitted if such combinations produce a stable compound.

[0078] When the number of a linking group is 0, such as -(CRR)0-, it indicates that the linking group is a single bond.

[0079] When one of the variables is selected as a single bond, it means that the two groups it connects to are directly connected. For example, when L in ALZ represents a single bond, it means that its structure is actually AZ.

[0080] When the listed linking groups do not specify their linking direction, the linking direction is arbitrary, for example, The linking group L is at this time The benzene ring and cyclopentyl group can be connected in the same direction as the reading order from left to right to form the structure. Alternatively, the phenyl and cyclopentyl groups can be connected in the reverse order of reading from left to right to form the phenyl group. The combination of linking groups, substituents, and / or their variants is permitted only if such a combination produces a stable compound.

[0081] Unless otherwise specified, the number of atoms in a ring is usually defined as the elemental number of the ring. For example, a “3-7 elemental ring” refers to a “ring” with 3 to 7 atoms arranged around it.

[0082] Unless otherwise specified, the term "halogen" refers to fluorine, chlorine, bromine, and iodine.

[0083] Unless otherwise specified, the term "C" 1-6 "alkyl" is used to denote a straight-chain or branched saturated hydrocarbon group consisting of 1 to 6 carbon atoms. The C 1-6 Alkyl groups include C 1-5 C 1-4 C 1-3 C 1-2 C 2-6 C 2-4 C6 and C5 alkyl groups, etc.; they can be monovalent (e.g., methyl), divalent (e.g., methylene), or polyvalent (e.g., methine). 1-6 Examples of alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), butyl (including n-butyl, isobutyl, s-butyl and t-butyl), pentyl (including n-pentyl, isopentyl and neopentyl), hexyl, etc.

[0084] Unless otherwise specified, the term "C" 1-3 "alkyl" is used to denote a straight-chain or branched saturated hydrocarbon group consisting of 1 to 3 carbon atoms. The C 1-3 Alkyl groups include C 1-2 and C 2-3 Alkyl groups, etc.; they can be monovalent (e.g., methyl), divalent (e.g., methylene), or polyvalent (e.g., methine). C 1-3 Examples of alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), etc.

[0085] Unless otherwise specified, the term "C" 2-6"Alkenyl" is used to denote a hydrocarbon group consisting of 2 to 6 carbon atoms, either straight-chain or branched, containing at least one carbon-carbon double bond. The carbon-carbon double bond can be located at any position within the group. 2-6 Alkenes include C 2-4 C 2-3 C4, C3, C2 alkenyl groups, etc.; they can be monovalent, divalent, or polyvalent. 2-6 Examples of alkenyl groups include, but are not limited to, vinyl, propenyl, butenyl, pentenyl, hexenyl, butadienyl, and pentadienyl.

[0086] Unless otherwise specified, the term "C" 1-6 "Alkoxy" refers to alkyl groups containing 1 to 6 carbon atoms that are attached to other parts of a molecule by an oxygen atom. The C 1-6 Alkyl groups include C 1-4 C 1-3 C 1-2 C 2-6 C 2-4 C6, C5, C4, and C3 alkoxy groups, etc.; C 1-6 Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), butoxy (including n-butoxy, isobutoxy, s-butoxy and t-butoxy), pentoxy (including n-pentoxy, isopentoxy and neopentoxy), hexoxy, etc.

[0087] Unless otherwise specified, the term "C" 1-6 "Alkylamino" refers to alkyl groups containing 1 to 6 carbon atoms that are attached to other parts of a molecule via an amino group. The C 1-6 Alkyl groups include C 1-4 C 1-3 C 1-2 C 2-6 C 2-4 C6, C5, C4, C3, and C2 alkylamino groups, etc.; C 1-6 Examples of alkylamino groups include, but are not limited to, -NHCH3, -N(CH3)2, -NHCH2CH3, -N(CH3)CH2CH3, -N(CH2CH3)2, -NHCH2CH2CH3, -NHCH(CH3)2, -NHCH2CH2CH2CH3, etc.

[0088] Unless otherwise specified, the term "C" 3-6 "Cycloalkyl" refers to a saturated cyclic hydrocarbon group consisting of 3 to 6 carbon atoms, which can be monocyclic or bicyclic systems. 3-6 Cycloalkyl groups include C 3-5 C 4-5 and C 5-6 Cycloalkyl groups, etc.; they can be monovalent, divalent, or polyvalent. C3-6 Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, etc.

[0089] Unless otherwise specified, the term "3-8 membered heterocyclic alkyl" on its own or in combination with other terms refers to a saturated monocyclic group consisting of 3 to 8 ring atoms, wherein 1, 2, 3, or 4 of the ring atoms are heteroatoms independently selected from O, S, and N, and the remainder are carbon atoms, wherein the nitrogen atom is optionally quaternized, and the nitrogen and sulfur heteroatoms may optionally be oxidized (i.e., NO and S(O)). z (z is 1 or 2). Furthermore, regarding the "3-8 membered heterocyclic alkyl", the heteroatom can occupy the connection position between the heterocyclic alkyl group and the rest of the molecule. The 3-8 membered heterocyclic alkyl group includes 4-6, 5-6, 7-8, 4, 5, and 6 membered heterocyclic alkyl groups, etc. Examples of 3-6 membered heterocyclic alkyl groups include, but are not limited to, nitrogen-containing heterocyclic butyl, oxo-heterocyclic butyl, thiocyclic butyl, pyrrolidinyl, pyrazolyl, imidazoalkyl, tetrahydrothiopheneyl, tetrahydrofuranyl (including tetrahydrofuran-2-yl), piperidinyl, tetrahydropyranyl, tetrahydrothiaranyl, piperazinyl, morpholinyl, thiomorpholinyl, etc.

[0090] Unless otherwise specified, the terms "6-10-membered aromatic ring" and "6-10-membered aryl" are used interchangeably, and the term "6-10-aryl" refers to a monovalent aromatic carbon ring system containing 6-10 carbon atoms and having at least one aromatic ring or at least one of the rings being aromatic rings. Examples of aryl groups are, but are not limited to, phenyl, naphthyl, biphenyl, or indenyl.

[0091] Unless otherwise specified, the terms "5-10-membered heteroaryl ring" and "5-10-membered heteroaryl" are used interchangeably in this invention. The term "5-10-membered heteroaryl" refers to a cyclic group consisting of 5 to 10 ring atoms with a conjugated π-electron system, wherein 1, 2, 3, or 4 of the ring atoms are heteroatoms independently selected from O, S, and N, and the remainder are carbon atoms. It can be a monocyclic or fused bicyclic system, wherein at least one ring in the system is aromatic. The nitrogen atom is optionally quaternized, and the nitrogen and sulfur heteroatoms may optionally be oxidized (i.e., NO and S(O)). z(z is 1 or 2). 5-10-membered heteroaryl groups can be attached to the rest of the molecule via heteroatoms or carbon atoms, including 5-8-membered, 5-7-membered, 5-6-membered, 5-membered, and 6-membered heteroaryl groups, etc. Examples of the 5-10 membered heteroaryl groups include, but are not limited to, pyrrole (including N-pyrrole, 2-pyrrole, and 3-pyrrole), pyrazolyl (including 2-pyrazolyl and 3-pyrazolyl), imidazole (including N-imidazolyl, 2-imidazolyl, 4-imidazolyl, and 5-imidazolyl), oxazolyl (including 2-oxazolyl, 4-oxazolyl, and 5-oxazolyl), triazolyl (1H-1,2,3-triazolyl, 2H-1,2,3-triazolyl, 1H-1,2,4-triazolyl, and 4H-1,2,4-triazolyl), tetrazolyl, isoxazolyl (including 3-isooxazolyl, 4-isooxazolyl, and 5-isooxazolyl), and thiazolyl (including 2-thiazolyl, 4-isooxazolyl, 4-isooxazolyl, and 5-isooxazolyl). α-thiazolyl and 5-thiazolyl, etc., furanyl (including 2-furanyl and 3-furanyl), thienyl (including 2-thienyl and 3-thienyl), pyridyl (including 2-pyridyl, 3-pyridyl and 4-pyridyl), pyrazinyl, pyrimidinyl (including 2-pyrimidinyl and 4-pyrimidinyl), benzothiazolyl (including 2-benzothiazolyl), purine, benzimidazolyl (including 2-benzimidazolyl), benzoxazolyl, indazole (including 5-indazole), isoquinolinyl (including 1-isoquinolinyl and 5-isoquinolinyl), quinoxalinyl (including 2-quinoxalinyl and 5-quinoxalinyl), or quinolinyl (including 3-quinolinyl and 6-quinolinyl). etc.

[0092] Unless otherwise specified, the terms "5-6 membered heteroaryl" and "5-6 membered heteroaryl" are used interchangeably in this invention. The term "5-6 membered heteroaryl" refers to a cyclic group consisting of 5 to 6 ring atoms with a conjugated π-electron system, wherein 1, 2, 3, or 4 of the ring atoms are heteroatoms independently selected from O, S, and N, and the remainder are carbon atoms. The nitrogen atom is optionally quaternized, and the nitrogen and sulfur heteroatoms may optionally be oxidized (i.e., NO and S(O)). z(z is 1 or 2). The 5-6 membered heteroaryl group can be attached to the rest of the molecule via a heteroatom or carbon atom. The 5-6 membered heteroaryl group includes 5-membered and 6-membered heteroaryl groups, etc. Examples of the 5-6 membered heteroaryl group include, but are not limited to, pyrrole (including N-pyrrole, 2-pyrrole, and 3-pyrrole), pyrazolyl (including 2-pyrazolyl and 3-pyrazolyl), imidazole (including N-imidazolyl, 2-imidazolyl, 4-imidazolyl, and 5-imidazolyl), oxazolyl (including 2-oxazolyl, 4-oxazolyl, and 5-oxazolyl), and triazolyl (1H-1,2,3-triazolyl, 2H-1,2,3-triazolyl, 1H-1,2,4-triazolyl). And 4H-1,2,4-triazolyl), tetrazolyl, isoxazolyl (including 3-isooxazolyl, 4-isooxazolyl and 5-isooxazolyl, etc.), thiazolyl (including 2-thiazolyl, 4-thiazolyl and 5-thiazolyl, etc.), furanyl (including 2-furanyl and 3-furanyl, etc.), thiophene (including 2-thiophene and 3-thiophene, etc.), pyridyl (including 2-pyridyl, 3-pyridyl and 4-pyridyl, etc.), pyrazinyl, pyrimidinyl (including 2-pyrimidinyl and 4-pyrimidinyl, etc.), etc.

[0093] Unless otherwise specified, C n-n+m Or C n -C n+m This includes any specific case with n to n+m carbons, such as C 1-7 This includes C1, C2, C3, C4, C5, C6, and C7, as well as any range from n to n+m, such as C 1-7 Including C 1-3 C 1-6 C 3-6 C 4-7 and C 5-7 Similarly, n-membered to n+m-membered rings represent the number of atoms in the ring from n to n+m. For example, 3-7-membered rings include 3-membered, 4-membered, 5-membered, 6-membered, and 7-membered rings, and also include any range from n to n+m. For example, 3-7-membered rings include 3-6-membered rings, 4-7-membered rings, 5-7-membered rings, and 6-7-membered rings, etc.

[0094] Preparation method

[0095] This invention also relates to a method for producing compounds of general formula (I) as defined above. In some embodiments, the methods for synthesizing the compounds of this application are as follows:

[0096]

[0097] When X is selected from halogens other than Br, such as F, Cl, I, etc., the method includes the following steps:

[0098] a) Using a brominating agent to bromate a compound of formula Ia to generate a compound of formula Ib, preferably, the brominating agent includes, but is not limited to, N-bromosuccinimide (NBS) or 1,3-dibromo-5,5-dimethylhydantoin, preferably, the reaction temperature is 0-30°C.

[0099] b) In the compound of formula Ib, one Br atom is replaced by another halogen atom to obtain a compound of formula Ic. Preferably, step b) uses... The reaction is carried out at low temperature with a strong base of organometallic salt, wherein X is a halogen other than Br, such as F, Cl, I, more preferably, wherein the strong base of organometallic salt is n-butyllithium, and preferably, the reaction temperature is -78°C.

[0100] c) Using an oxidizing agent to oxidize the compound of formula Ic to the compound of formula Id, preferably, the oxidizing agent includes, but is not limited to, K2Cr2O7, and preferably, the reaction temperature is 60-150°C;

[0101] d) Compounds of formula Id with H2N-LR 1 The reaction yields a compound of formula Ie, preferably at a temperature of 50-150°C;

[0102] e) Combine the compound of formula Ie with R 2 R 3 The reaction is carried out by NH to give a compound of formula I. Preferably, the reaction is carried out in the presence of the catalyst bis(triphenylphosphine)palladium(II) chloride. Preferably, the reaction temperature is 20-150°C.

[0103] When X is selected from Br, the method includes:

[0104] a) Using a brominating agent to bromate a compound of formula Ia to generate a compound of formula Ib, preferably, the brominating agent includes, but is not limited to, N-bromosuccinimide (NBS), preferably, the reaction temperature is 0-30°C;

[0105] c) Using an oxidizing agent to oxidize the compound of formula Ib to the compound of formula Id, preferably, the oxidizing agent includes, but is not limited to, K2Cr2O7, and preferably, the reaction temperature is 60-150°C;

[0106] d) The compound of formula Id reacts with H2N-L-R1 to give the compound of formula Ie. Preferably, the reaction temperature is 50-150℃.

[0107] e) Combine the compound of formula Ie with R 2 R 3The reaction is carried out by NH to give a compound of formula I. Preferably, the reaction is carried out in the presence of the catalyst bis(triphenylphosphine)palladium(II) chloride. Preferably, the reaction temperature is 20-150°C.

[0108]

[0109] Alternatively, when X is selected from halogens other than Br, such as F, Cl, I, etc., the method includes the following steps:

[0110] f) Using a brominating agent to bromate the compound of formula Ia to generate the compound of formula Ib, preferably, the brominating agent includes, but is not limited to, N-bromosuccinimide (NBS) or 1,3-dibromo-5,5-dimethylhydantoin, preferably, the reaction temperature is 0-30°C.

[0111] g) In the compound of formula Ib, both Br atoms are replaced by other halogen atoms to obtain the compound of formula Ic. Preferably, step b) uses... The reaction is carried out at low temperature with a strong base of organometallic salt, wherein X is a halogen other than Br, such as F, Cl or I, more preferably, wherein the strong base of organometallic salt is n-butyllithium, and preferably, the reaction temperature is -78°C.

[0112] h) Using an oxidizing agent to oxidize the compounds of formulas I-c' to the compounds of formulas I-d', preferably, the oxidizing agent includes, but is not limited to, K2Cr2O7, and preferably, the reaction temperature is 60-150°C;

[0113] i) Compounds of formula I-d' and H2N-LR 1 The reaction yields compounds of formula I-e', preferably at a reaction temperature of 50-150°C;

[0114] j) The compound of formula I-e' is combined with R 2 R 3 The reaction is carried out by NH to give a compound of formula I. Preferably, the reaction is carried out in the presence of the catalyst bis(triphenylphosphine)palladium(II) chloride. Preferably, the reaction temperature is 20-150°C.

[0115] As is known to those skilled in the art, certain active groups (e.g., -NH2, -OH, etc.) require routine protection and deprotection when necessary, which is familiar to those skilled in the art.

[0116] Products and their applications

[0117] This disclosure also provides a product, which may be, for example, a pharmaceutical, pharmaceutical composition, or kit / pharmaceutical, containing an effective amount of the hnRNPA2B1 agonist compound of this disclosure, and a pharmaceutically or immunologically acceptable carrier. As used herein, the terms “active substance” and “active substance of this disclosure” are used interchangeably and refer to a hnRNPA2B1 agonist compound having the structural formula (I) or a derivative thereof or product thereof.

[0118] As used herein, the term "pharmaceuticalally acceptable" refers to a substance that is suitable for use in humans and / or animals without excessive adverse effects (such as toxicity, irritation, and allergic reactions), i.e., a reasonable benefit / risk ratio. As used herein, the term "effective amount" refers to an amount that is functional or active in humans and / or animals and is acceptable to humans and / or animals.

[0119] As used herein, the term "pharmaceuticalally acceptable carrier" refers to a carrier used for the administration of therapeutic agents, including various excipients and diluents. This term refers to pharmaceutical carriers that are not essential active ingredients themselves and do not cause excessive toxicity upon administration. Suitable carriers are well known to those skilled in the art, and a thorough discussion of pharmaceutically acceptable excipients can be found in Remington's Pharmaceutical Sciences (Mack Pub. Co., NJ 1991).

[0120] Pharmaceutically acceptable carriers in the composition may contain liquids such as water, saline, glycerol, and ethanol. Additionally, these carriers may contain auxiliary substances such as fillers, disintegrants, lubricants, glidants, effervescent agents, wetting agents or emulsifiers, flavoring agents, pH buffers, etc. Typically, these substances are formulated in a non-toxic, inert, and pharmaceutically acceptable aqueous carrier medium, with a pH usually around 5-8, preferably around 6-8.

[0121] The active substance in the product disclosed herein accounts for 0.001 to 99.9 wt% of the total weight of the composition; preferably 1 to 95 wt% of the total weight of the composition, more preferably 5 to 90 wt%, and more preferably 10 to 80 wt%. The balance consists of pharmaceutically acceptable carriers and other additives.

[0122] As used herein, the term "unit dosage form" refers to a dosage form in which the products of this disclosure are prepared for a single dose for ease of administration, including but not limited to various solid dosage forms (such as tablets), liquid dosage forms, capsules, and sustained-release formulations.

[0123] In another preferred embodiment of this disclosure, the product is a single dosage form or multiple dosage forms, and the content of the active substance is 0.01–2000 mg / dose, preferably 0.1–1500 mg / dose, more preferably 1–1000 mg / dose. In another preferred example of this disclosure, 1–6 doses of the composition of this disclosure are administered daily, preferably 1–3 doses; most preferably, the daily dose is 1 dose.

[0124] It should be understood that the effective dose of active substances such as hnRNPA2B1 agonist compounds can vary depending on the severity of the condition of the patient being treated. The specific dosage depends on the individual patient's circumstances (e.g., weight, age, physical condition, and desired outcome), and is within the judgment of a skilled physician.

[0125] The products disclosed herein may be in solid form (e.g., granules, tablets, lyophilized powder, suppositories, capsules, sublingual tablets) or liquid form (e.g., oral liquid) or other suitable forms. Routes of administration may include, but are not limited to: (1) conventional administration methods, such as gastrointestinal administration (e.g., oral administration), non-gastrointestinal administration (e.g., solution injection, intravenous infusion), such as mucosal administration, transdermal administration, respiratory nebulization, nasal drops, sprays, oral administration, intramuscular injection and / or intravenous administration, etc.; (2) linking hnRNPA2B1 agonists with transferrin / poly-L-lysine complexes to enhance their biological effects; (3) encapsulating drugs in liposomes to mediate entry into cells, which facilitates the smooth entry of compound molecules and protects them from hydrolysis by various extracellular enzymes; (4) using liposomes to transport active substances to their specific target tissues and cells.

[0126] In addition, the products disclosed herein may also contain other active substances for improving and treating infectious diseases. These other active substances include, but are not limited to, one or more of the following clinically commonly used antibiotics: β-lactams (penicillins and cephalosporins), aminoglycosides, tetracyclines, chloramphenicol, macrolides, antifungal antibiotics, and antituberculosis antibiotics.

[0127] In some embodiments, other active substances that modulate anti-infection are administered before, simultaneously with, or after the product of this disclosure. These other active substances have activity in preventing or treating diseases associated with infection (especially viral infection), infection-induced damage, infection-induced chronic inflammatory diseases, and / or their symptoms.

[0128] In some embodiments, activation of hnRNPA2B1 by the compounds or products of this application to further modulate the immune system provides treatment for diseases, including those caused by exogenous factors. Exemplary infections caused by exogenous factors that can be treated and / or prevented by the methods of this invention include bacterial infections (e.g., Gram-positive or Gram-negative bacteria), fungal infections, parasitic infections, and viral infections. In one embodiment of the invention, the infection is a bacterial infection (e.g., infection with Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Salmonella spp., Staphylococcus aureus, Streptococcus, or vancomycin-resistant enterococci) or sepsis. In other embodiments, the infection is a fungal infection (e.g., infection with molds, yeasts, or higher fungi). In other embodiments, the infection is a parasitic infection (e.g., an infection caused by single-celled or multi-celled parasites, including Giardia duodenalis, Cryptosporidium parvum, Cyclospora cayetanensis, and Toxoplasma gondii). In still other embodiments, the infection is a viral infection (e.g., AIDS-related viral infections, avian influenza, chickenpox, cold sores, the common cold, gastroenteritis, glandular fever, influenza, measles, mumps, pharyngitis, pneumonia, rubella, SARS, and lower respiratory tract infections or upper respiratory tract infections (e.g., respiratory syncytial virus).

[0129] In a preferred embodiment, the product can be used to prevent or treat diseases associated with infection (especially viral infection), chronic inflammatory diseases caused by infection, and / or their symptoms; for example, the pharmaceutical compositions disclosed herein can be used to prevent or treat viral infectious diseases known in the art to be treatable or preventable, such as tissue damage caused by viral infection; inflammatory damage to organs; multiple organ failure.

[0130] In some embodiments, the infections that can be treated or prevented by the compounds of this application are DNA-involved and / or mediated infections. In some embodiments, the infection is a viral infection or an infection caused by DNA-involved bacteria, fungi, or a combination thereof, such as a DNA viral infection, like an infection caused by one or more viruses selected from the group consisting of: herpes simplex virus, hepatitis B virus, adenovirus, poxvirus, microDNA virus, adeno-associated virus, coronavirus, influenza virus, rhinovirus, parainfluenza virus, respiratory syncytial virus, coxsackie virus, echovirus, and novel enterovirus.

[0131] In some embodiments, the infection-related disease and / or symptoms are selected from one or more of the following groups: pathological damage caused by infection; insufficient or excessive production of cytokines such as interferon after infection; endotoxic shock or death; inflammatory damage to organs; multiple organ failure, for example, the organs are selected from: liver, spleen, brain, kidney, heart, lung, stomach, intestine; chronic inflammatory diseases caused by viral infection (e.g., autoimmune diseases such as inflammatory bowel disease, rheumatoid arthritis, systemic lupus erythematosus, chronic nephritis, tuberculosis, chronic gastrointestinal diseases). Preferably, the chronic inflammatory diseases and / or symptoms caused by viral infection include: autoimmune diseases such as inflammatory bowel disease, rheumatoid arthritis, systemic lupus erythematosus, chronic nephritis, tuberculosis, chronic gastrointestinal diseases. In some embodiments, the virus is selected from HSV, HBV, and VSV.

[0132] In some embodiments, the compounds or products of this application are administered prophylactically as preventative agents before an infection (especially a pathogen infection) occurs to prevent the infection from occurring or to reduce the severity of subsequent infections. In some embodiments, the compounds or products of this application are administered as therapeutic agents after an infection occurs to reduce the severity of the infection and disease. In some embodiments, the compounds or products of this application are administered both as prophylactic and therapeutic agents, either continuously or intermittently before and after an infection occurs.

[0133] Those skilled in the art can combine the foregoing technical solutions and features in any way without departing from the inventive concept and protection scope of this invention. Other aspects of this invention will be apparent to those skilled in the art from the disclosure herein.

[0134] Example

[0135] The present disclosure is further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the disclosure. Those skilled in the art can make appropriate modifications and variations to the present disclosure, and such modifications and variations are all within the scope of the present disclosure.

[0136] Experimental methods not specifically described in the following examples can be performed using conventional methods in the art, such as those described in *Molecular Cloning: A Laboratory Manual* (3rd edition, New York: Cold Spring Harbor Laboratory Press, 1989) or according to the conditions recommended by the supplier. DNA sequencing methods are conventional in the art and can also be provided by commercial companies.

[0137] Unless otherwise stated, percentages and parts are by weight. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as are familiar to one skilled in the art. Furthermore, any methods and materials similar or equivalent to those described herein may be used in the methods of this disclosure. The preferred embodiments and materials described herein are for illustrative purposes only.

[0138] I. Compound Synthesis and Characterization Examples

[0139] Example I.1: Synthesis of compound A (FPC-1):

[0140]

[0141] *eq equivalent

[0142] 1-1. Preparation of Compound 2

[0143]

[0144] NBS (57.5 g, 320 mmol) was added to DMF (120 mL) and cooled to 0 °C. Compound 1 (20 g, 130 mmol) was added, and the resulting suspension was stirred and allowed to warm naturally to room temperature overnight. The reaction solution was filtered, and the filter cake was washed three times with ethanol. The dried filter cake was collected and recrystallized with EA to give the pure product (10.1 g, off-white crystals).

[0145] 1 ¹H NMR (300MHz, chloroform-d) δ 7.79 (d, J = 7.4Hz, 2H), 7.09 (d, J = 8.8Hz, 2H), 3.30 (s, 4H).

[0146] 1-2. Preparation of Compound 3

[0147]

[0148] Compound 2 (6 g, 19.2 mol) was dissolved in 300 mL of anhydrous THF and cooled to -80 °C. Then, n-butyllithium (9.2 mL, 2.5 M) was added dropwise, and the reaction mixture was stirred at the same temperature for at least one hour. After the slow addition of N-fluorobis(benzenesulfonamide) (9 g, 28.8 mmol) in 100 mL of anhydrous THF, the mixture was stirred at -80 °C for another 60 minutes. The reaction mixture was then heated to room temperature overnight and poured into an NH4Cl solution. The aqueous phase was then extracted with DCM (3 × 50 mL), the organic phases were combined, and dried over MgSO4. The solvent was removed under low pressure. The product was purified by column chromatography using n-hexane as the eluent to give compound 3 (4.1 g, white solid).

[0149] 1 ¹H NMR (300MHz, chloroform-d) δ 7.66 (d, J = 7.4Hz, 1H), 7.21–7.09 (m, 3H), 3.35 (s, 4H).

[0150] 1-3. Preparation of Compound 4

[0151]

[0152] Compound 3 (4.5 g, 17.9 mmol) was added to a solution of potassium dichromate (24 g, 82.4 mmol) in glacial acetic acid (50 mL). The solution was heated under reflux for 16 hours and then poured into ice water. The resulting precipitate was collected by filtration and washed with water. The filter cake was air-dried, added to DCM and refluxed for 30 minutes, filtered, washed with DCM, and the filter cake was added to DCM and refluxed for 15 minutes, filtered, and washed with DCM. All filtrates were combined and concentrated under vacuum to give compound 4 (4.1 g, crude product, yellow solid).

[0153] 1-4. Preparation of INT-1

[0154]

[0155] N-Boc-ethylenediamine (3.3 g, 20.8 mmol) was added to a 100 mL solution of compound 4 (4.1 g, 13.9 mmol) in EtOH, and the mixture was heated under reflux for 1 hour. The reaction mixture was concentrated, and the residue was purified by column chromatography (PE / EA = 5 / 1) to give a white solid (2.8 g).

[0156] 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).

[0157] 1-5. Preparation of FPC-1-1

[0158]

[0159] Cesium carbonate (5.55 g, 17 mmol) and 1-(2-pyridone ethyl)piperazine (1.05 g, 5.7 mmol) were added to a toluene solution (175 mL) of INT-1 (2.5 g, 5.7 mol). Bis(triphenylphosphine)palladium(II) chloride (400 mg, 0.57 mmol) was added to the mixture under argon protection. The reaction was carried out at 80 °C for 16 hours. The reaction solution was cooled to room temperature, water was added, and the mixture was stirred. The organic phase was separated, and the aqueous phase was extracted once with DCM. The combined organic phases were dried over anhydrous magnesium sulfate. The crude product was purified by Prep-TLC (DCM / MeOH = 10 / 1 + NH3·H2O) by vacuum concentration and column chromatography to give 240 mg of a pure yellow solid.

[0160] 1 ¹H NMR (400MHz, chloroform-d) δ 8.56 (dd, J = 7.7, 4.3Hz, 1H), 8.51 (d, J = 8.3Hz, 1H), 7.34–7.27 (m, 1H), 7.17 (d, J = 8.2Hz, 1H), 4.99 (s, 1H), 4.31 (s, 2H), 3.79–2.45 (m, 18H), 2.18 (s, 4H), 1.28 (s, 9H).

[0161] 1-6. Preparation of FPC-1

[0162]

[0163] Compound FPC-1-1 (240 mg, 0.44 mmol) was dissolved in 11 mL of DCM / MeOH (10 / 1). 5 M HCl / 1,4-dioxane (5 mL) was added to the mixture and the mixture was stirred at room temperature for 2 hours. The mixture was then concentrated under vacuum. The residue was dissolved in a small amount of methanol, precipitated with diethyl ether, centrifuged, washed with diethyl ether, centrifuged again, and dried under vacuum to give 240 mg of the pure product (yellow solid).

[0164] 1 ¹H NMR (300MHz, methanol-d⁴) δ 8.67–8.58 (m, 2H), 7.56 (dd, J = 13.2, 8.3 Hz, 1H), 7.47 (d, J = 8.3 Hz, 1H), 4.49–4.42 (m, 2H), 4.02–3.69 (m, 10H), 3.69–3.38 (m, 8H), 2.18 (s, 4H).

[0165] MS(ESI)m / z[M+H] + =440.3.

[0166] Example I.2: Synthesis of Compound B (FPC-2)

[0167]

[0168] 2-1. Preparation of FPC-2-1

[0169]

[0170] 10 mL of ethylene glycol monomethyl ether was added to a single-necked flask, followed by INT-1 (110 mg, 0.25 mmol) and 3-(piperazin-1-yl)prop-1-ol (44 mg, 0.3 mmol). The reaction mixture was stirred at 100 °C. After 2 hours, the solvent was removed under vacuum, and the residue was purified by TLC to give 20 mg of a yellow solid.

[0171] 1 ¹H NMR (400MHz, chloroform-d) δ 8.63–8.49 (m, 2H), 7.31 (dd, J = 12.8, 8.3Hz, 1H), 7.18 (d, J = 8.3Hz, 1H), 4.97 (s, 1H), 4.42–4.23 (m, 2H), 3.87 (t, J = 5.0Hz, 2H), 3.60–3.28 (m, 8H), 2.92 (s, 4H), 1.89 (s, 2H), 1.29 (s, 9H).

[0172] 2-2. Preparation of FPC-2

[0173]

[0174] Compound FPC-2-1 (20 mg, mmol) was dissolved in 2 mL of DCM / MeOH (1 / 1). 5 M HCl / 1,4-dioxane (2 mL) was added, and the mixture was stirred at room temperature for 2 hours. The mixture was then concentrated under vacuum, and the residue was dissolved in a small amount of methanol, precipitated with diethyl ether, centrifuged, washed with diethyl ether, centrifuged again, and dried under vacuum to give 5 mg of the pure product (yellow solid).

[0175] 1 ¹H NMR (400MHz, methanol-d⁴) δ 8.72–8.46 (m, 2H), 7.50 (ddd, J = 26.1, 12.4, 8.3 Hz, 2H), 4.45 (t, J = 5.6 Hz, 2H), 3.93–3.69 (m, 6H), 3.45 (dt, J = 15.0, 9.8 Hz, 5H), 3.34 (s, 3H), 2.07 (dt, J = 12.2, 6.1 Hz, 2H).

[0176] MS(ESI)m / z[M+H] + =401.47.

[0177] Example I.3: Synthesis of Compound C (FPC-3)

[0178]

[0179] 3-1. Preparation of FPC-3-1

[0180]

[0181] Cesium carbonate (1.78 g, 5.48 mmol) and N-ethyl-2-(pyrrolidone-1-yl)ethyl-1-amine (286 mg, 2.01 mmol) were added to a toluene solution (70 mL) of INT-1 (800 mg, 1.82 mmol). Bis(triphenylphosphine)palladium(II) chloride (128 mg, 0.18 mmol) was added to the mixture under argon protection. The reaction was carried out at 80 °C for 16 hours. The reaction solution was cooled to room temperature, water was added, and the mixture was stirred. The organic phase was separated, and the aqueous phase was extracted once with DCM. The obtained organic phases were combined and dried over anhydrous magnesium sulfate. The crude product was purified by Prep-TLC (DCM / MeOH = 10 / 1 + NH3·H2O) by vacuum concentration and column purification to give 30 mg of a pure yellow solid.

[0182] 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).

[0183] 3-2. Preparation of FPC-3

[0184]

[0185] Compound FPC-3-1 (30 mg, 0.06 mmol) was dissolved in 4.4 mL LCM / MeOH (10 / 1). 5 M HCl / 1,4-dioxane (2 mL) was added, and the mixture was stirred at room temperature for 2 hours. The mixture was then concentrated under vacuum, and the residue was dissolved in a small amount of methanol, precipitated with diethyl ether, centrifuged, washed with diethyl ether, centrifuged again, and dried under vacuum to give 25 mg of the pure product (yellow solid).

[0186] 1 H NMR (300MHz, methanol-d4) δ8.67-8.54(m,2H),7.58-7.46(m,2H),4.51-4.41(m,2H),3.87-3.77(m,2H),3.77-3.65(m ,2H),3.51(q,J=6.6Hz,4H),3.34(d,J=1.8Hz,1H),3.20-3.05(m,2H),2.20-1.92(m,5H),1.10(t,J=7.0Hz,3H).

[0187] MS(ESI)m / z[M+H] + =399.47.

[0188] Example I.4: Synthesis of Compound D (FPC-5)

[0189]

[0190] 4-1. Preparation of FPC-5-1

[0191]

[0192] Cesium carbonate (460 mg, 1.41 mmol) and N,2-dimethylpropyl-1-amine (49 mg, 0.56 mmol) were added to a toluene solution (20 mL) of INT-1 (205 mg, 0.47 mmol). Bis(triphenylphosphine)palladium(II) chloride (32 mg, 0.045 mmol) was added to the mixture under argon protection. The reaction was carried out at 80 °C for 16 hours. The reaction solution was cooled to room temperature, water was added and stirred, and the organic phase was separated. The aqueous phase was extracted once with DCM. The combined organic phases were dried over anhydrous magnesium sulfate. The crude product was purified by vacuum concentration and column chromatography (DCM / MeOH = 10 / 1 + NH3·H2O) to give 10 mg of a pure yellow solid.

[0193] 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).

[0194] 4-2. Preparation of FPC-5

[0195]

[0196] Compound FPC-5-1 (10 mg, 0.022 mmol) was dissolved in 1.1 mL of DCM / MeOH (10 / 1). 5 M HCl / 1,4-dioxane (0.5 mL) was added, and the mixture was stirred at room temperature for 2 hours. The mixture was then concentrated under vacuum, and the residue was dissolved in a small amount of methanol, precipitated with diethyl ether, centrifuged, washed with diethyl ether, centrifuged again, and dried under vacuum to give 9 mg of the pure product (yellow solid).

[0197] 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).

[0198] MS(ESI)m / z[M+H] + =344.42.

[0199] Example I.5: Synthesis of compound E (AIR-2)

[0200]

[0201] 5-1. Preparation of Compound 5

[0202]

[0203] Compound 3 (31.90 g, 127.04 mmol) was dissolved in anhydrous THF (800 mL). The reaction mixture was purged three times with argon and then cooled to -78 °C. Then, n-BuLi (60.98 mL, 2.5 M / L, 152.45 mmol) was added dropwise, and the reaction mixture was stirred at the same temperature for at least 10 minutes. N-fluorobis(benzenesulfonamide) (NFSI) (60.09 g, 190.57 mmol) in 200 mL of anhydrous THF was added dropwise, and the mixture was stirred at -78 °C for 1 hour. The reaction mixture was then allowed to warm to room temperature naturally and stirred overnight. The reaction mixture was quenched with NH4Cl solution and then extracted twice with EA. The combined organic layers were washed with water and brine, dried over Na2SO4, and filtered. The filtrate was concentrated to give the crude product, which was added to DCM and stirred. The precipitate was filtered, the filter cake was washed twice with DCM, all filtrates were combined and concentrated under vacuum to obtain a crude product, which was then purified by column chromatography (PE) to give compound 5 as a white solid (15.42 g, 63.82%).

[0204] 1 H NMR (300MHz, Chloroform-d) δ7.16 (d, J = 7.6Hz, 2H), 7.12–7.04 (m, 2H), 3.37 (s, 4H).

[0205] 5-2. Preparation of INT-2

[0206]

[0207] Compound 5 (15.42 g, 81.08 mmol) was added to a solution of potassium dichromate (109.72 g, 372.96 mmol) in glacial acetic acid (220 mL). The solution was stirred at 80 °C for 12 hours. The reaction mixture was quenched with EA and water, and extracted twice with EA. The combined organic layers were washed with water and brine, dried over Na₂SO₄, and filtered. The filtrate was concentrated to give the crude product, which was then added to PE and an ultrasonic dispersion. The resulting precipitate was filtered and dried to obtain crude INT-2 (10.55 g) as a brownish-yellow solid, which was ready for use without further purification.

[0208] 5-3. Preparation of AIR-2-1

[0209]

[0210] INT-2 (5 g, 21.35 mmol) was dissolved in EtOH (120 mL). The reaction system was purged three times with argon gas, and then heated to 50 °C. N2 was added. 1 N 1-Dimethylethane-1,2-diamine (2.79 mL, 25.62 mmol) was heated to 80 °C (reflux) for 4 hours. TLC showed the reaction was complete. The reaction mixture was concentrated, and the residue obtained was purified by column chromatography (DCM:MeOH = 30:1 to 10:1) to give a brownish-yellow solid AIR-2-1 (1.81 g, 27.86%). MS (ESI), m / z: 305.4 [M+H] + .

[0211] 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).

[0212] 5-4. Preparation of AIR-2

[0213]

[0214] AIR-2-1 (1.81 g, 5.95 mmol) was dissolved in 1,4-dioxane (150 mL), and the reaction system was purged three times with argon. 1-(2-pyridone-ethyl)piperazine (compound A) (1.20 g, 6.54 mmol) was added, and the mixture was stirred overnight at room temperature. The reaction mixture was concentrated, and the residue was purified by column chromatography (DCM:MeOH = 15:1 to 5:1) to give the pure product dissolved in DCM. Then, 5 M HCl / 1,4-dioxane (1.5 mL) was added and stirring continued, followed by precipitation with diethyl ether. The precipitate was filtered, washed with diethyl ether, and dried to give a yellow solid AIR-2 (1.21 g). MS (ESI), m / z: 468.47 [M+H] + .

[0215] 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).

[0216] Example I.6: Synthesis of compound F (AIR-3)

[0217]

[0218] 6-1. Preparation of Compound 2

[0219]

[0220] NBS (499.72 g, 2.81 mol) was added to DMF (1000 mL) and stirred until most of the solid dissolved. The reaction system was purged twice with argon and then cooled to 9 °C. Compound 1 (176 g, 1.14 mol) was added, and the mixture was stirred at 9 °C for 2 hours. The reaction mixture was then allowed to warm to room temperature naturally and stirred over the weekend. The reaction solution was filtered, and the filter cake was washed twice with PE / EA = 1 / 1. The dried filter cake was collected and recrystallized from EA to give pure compound 2 (102.57 g, 28.80%) as off-white crystals.

[0221] 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).

[0222] 6-2. Preparation of Compound 3

[0223]

[0224] Compound 2 (50 g, 160.26 mmol) was dissolved in anhydrous THF (950 mL). The reaction mixture was purged three times with argon and then cooled to -78 °C. Then, n-BuLi (76.92 mL, 2.5 M / L, 192.31 mmol) was added dropwise, and the reaction mixture was stirred at the same temperature for at least 20 minutes. N-fluorobis(benzenesulfonamide) (75.8 g, 240.38 mmol) was added dropwise to 250 mL of anhydrous THF, and the mixture was stirred at -78 °C for 1.5 hours. The reaction mixture was then allowed to warm to room temperature naturally and stirred overnight. The reaction mixture was quenched with NH4Cl solution and then extracted twice with EA. The combined organic layers were washed with water and brine, dried over Na2SO4, and filtered. The filtrate was concentrated to give the crude product, which was added to DCM and stirred. The precipitate obtained by filtration was washed twice with DCM, all filtrates were combined and concentrated under vacuum to obtain crude product, which was then purified by column chromatography (PE:EA = 50:1) to obtain compound 3 (31.90 g, 79.27%) as a white solid.

[0225] 1H NMR (300MHz, Chloroform-d) δ7.66 (d, J = 7.4Hz, 1H), 7.21–7.09 (m, 3H), 3.35 (s, 4H).

[0226] 6-3. Preparation of Compound 4

[0227]

[0228] Compound 3 (9.29 g, 37.00 mmol) was added to a solution of potassium dichromate (50.07 g, 170.19 mmol) in glacial acetic acid (100 mL). The solution was heated under reflux for 16 hours and then poured into ice water. The precipitate was collected by filtration and washed with water. The filter cake was air-dried, added to DCM and refluxed for 30 minutes, filtered, washed with DCM, and the filter cake was added to DCM and refluxed for 15 minutes, filtered, and washed with DCM. All filtrates were concentrated to give crude compound 3 (6.72 g), which could be used without further purification.

[0229] 6-4. Preparation of AIR-3-1

[0230]

[0231] To a 20 mL solution of compound 4 (400 mg, 1.36 mmol) in EtOH, DIPEA (358.48 μL, 2.17 mmol) and tert-butyl (2-aminoethyl)(methyl)carbamate hydrochloride (428.46 mg, 2.03 mmol) were added. The reaction mixture was purged with argon five times and then stirred (refluxed) at 80 °C for 2 hours. TLC showed that the reaction was complete. The reaction mixture was concentrated, and the residue was purified by column chromatography (PE:EA = 10:1 to 4:1) to give a yellow solid AIR-3-1 (244 mg, 39.88%).

[0232] 6-5. Preparation of AIR-3-2

[0233]

[0234] To a 15 mL solution (244 mg, 540.68 mmol) of AIR-3-1 in toluene, NaOt-Bu (103.92 mg, 1.08 mol), 1-(2-pyridone-ethyl)piperazine (118.92 mg, 648.81 mmol), and BINAP (101.00 mg, 162.20 mmol) were added. The reaction mixture was purged with argon five times, followed by the addition of Pd(OAc)₂ (24.28 mg, 108.14 mmol). The reaction mixture was purged with argon three times, and the reaction mixture was stirred at 90 °C for 12 hours under argon protection. The reaction mixture was quenched with water and extracted twice with EA. The combined organic layers were washed with water and brine, dried over Na₂SO₄, and filtered. The filtrate was concentrated to obtain the crude product, which was purified by column chromatography (DCM:MeOH = 100:1 to 10:1, with the addition of NH3·H2O) and Prep-TLC (DCM:MeOH = 10:1, with the addition of NH3·H2O) to obtain AIR-3-2 (40 mg, 13.36%).

[0235] 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).

[0236] 6-6. Preparation of AIR-3

[0237]

[0238] AIR-3-2 (40 mg, 72.24 mmol) in a DCM / MeOH = 10 / 1 solution (3 mL) was purged three times with argon, followed by the addition of 5 M HCl / 1,4-dioxane (1.5 mL). The resulting mixture was stirred at room temperature for 1.5 hours. TLC showed the reaction was complete. The reaction mixture was precipitated with diethyl ether, centrifuged, and the precipitate was washed with diethyl ether, centrifuged, and dried to give 33 mg of crude product. The crude product was further purified by Prep-TLC (DCM:MeOH = 20:1, with the addition of NH3·H2O) to give AIR-3 (10 mg) as a yellow solid. MS (ESI), m / z: 454.58 [M+H] +.HPLC 92.439% (220nm), HPLC 96.712% (254nm).

[0239] 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).

[0240] Example II. Activity Test

[0241] Example II.1: Testing the activation of the hnRNPA2B1 and TBK1-IRF3 pathways in immune cells by the compound.

[0242] Primary peritoneal macrophages were obtained from mice: C57BL / 6 mice (6-8 weeks old, female, purchased from Shanghai Bikai Laboratory Animal Co., Ltd.) were injected intraperitoneally with 2 ml of 3% mercaptoacetate (Sigma-Aldrich) solution once. Three days later, the mice were sacrificed by cervical dislocation, and the peritoneal cavity was flushed with serum-free culture medium. Cells were obtained by centrifugation after aspiration. Primary peritoneal macrophages were cultured in DMEM medium.

[0243] Primary peritoneal macrophages (cell density of 1 x 10^6 cells per well) were stimulated with 20 μM test compound E (prepared as in Example I.5, dissolved in physiological saline) for 30 minutes. Subcellular localization of hnRNPA2B1 was detected by immunofluorescence, and activation of TBK1 and IRF3 was detected by SDS-PAGE electrophoresis and immunoblotting.

[0244] The activation status of hnRNPA2B1, TBK1, and IRF3 is as follows: Figure 1 , 2 As shown.

[0245] The results showed that compound E significantly activated the nuclear export of macrophage hnRNPA2B1 cells and the activation of downstream TBK1 and IRF3. Furthermore, similar results were obtained when other compounds prepared in Example I were used in the same experiment.

[0246] The above results demonstrate that the tested compound activates the hnRNPA2B1 pathway and is an hnRNPA2B1 agonist.

[0247] Example II.2: Testing the broad induction of type I interferon production in multiple cell types by the compound

[0248] Primary mouse peritoneal macrophages (prepared as described in Example II.1) were cultured in DMEM medium, human THP1 monocytes (purchased from ATCC) were cultured in 1640 medium, and L929 fibroblasts were cultured in DMEM medium.

[0249] Cells were stimulated with 20 μM test compound E (cell density 2 x 10^5 cells per well) for 6 hours. Total RNA was extracted from the cells, and the transcription level of type I interferon (IFN-β) was detected by real-time quantitative PCR. Cell culture supernatant was collected 18 hours after stimulation, and the IFN-β protein level was detected by ELISA.

[0250] Test the activation of type I interferon by the compound, such as Figure 3 , 4 As shown in Figure 5.

[0251] The results showed that compound E significantly activated the expression of type I interferon in various cells. Furthermore, similar results were obtained when other compounds prepared in Example I were used in the same experiment.

[0252] The above results demonstrate that the tested compound can induce the production of type I interferon by activating the hnRNPA2B1 pathway, and is therefore a biologically active hnRNPA2B1 agonist. Thus, this disclosure defines such novel compounds as hnRNPA2B1 agonists.

[0253] Example II.3: hnRNPA2B1 agonist significantly induced the expression of multiple interferon-induced genes.

[0254] Primary peritoneal macrophages (prepared as described in Example II.1) were cultured in DMEM medium, and L929 fibroblasts were cultured in DMEM medium.

[0255] Cells were stimulated at a concentration of 20 μM (cell density of 2 x 10^5 cells per well) for 18 hours. Total RNA was extracted from the cells, and the expression of interferon-stimulated genes such as ISG15 and OAS1 was detected by real-time quantitative PCR.

[0256] The transcriptional level of interferon-stimulated genes, such as Figure 6 , 7 As shown.

[0257] The results showed that the hnRNPA2B1 agonist compound E significantly activated the expression of interferon-stimulated genes in both immune and non-immune cells. Furthermore, similar results were obtained when the same experiments were performed using other compounds prepared in Example I.

[0258] Example II.4: hnRNPA2B1 inhibits the replication of DNA virus HSV-1

[0259] L929 cells were cultured in DMEM medium and infected with HSV-1 (MOI, 1). Simultaneously, the cells were treated with either the hnRNPA2B1 agonist compound E (10 μM) or a negative control (physiological saline). Total RNA was collected from the cells at 24, 36, and 48 hours, and HSV-1 mRNA was detected by real-time quantitative PCR.

[0260] HSV-1 replication level such as Figure 8 As shown.

[0261] The results showed that the hnRNPA2B1 agonist compound E significantly inhibited the replication of the DNA virus HSV-1. Furthermore, similar results were obtained when the same experiments were performed using other compounds prepared in Example I.

[0262] Example II.5: hnRNPA2B1 agonist inhibits the replication of DNA virus HBV

[0263] HepG2.2.15 cells (cell density of 1 x 10^4 cells per well) were treated with either hnRNPA2B1 agonist compound E (10 μM) or a negative control (physiological saline). Total RNA was collected at 24, 36, and 48 hours, and HBV mRNA was detected by real-time quantitative PCR.

[0264] HBV replication level such as Figure 9 As shown.

[0265] The results showed that the hnRNPA2B1 agonist compound E significantly inhibited the replication of the DNA virus HBV. Furthermore, similar results were obtained when the same experiments were performed using other compounds prepared in Example I.

[0266] Example II.6: hnRNPA2B1 inhibits the replication of RNA virus VSV

[0267] L929 cells were cultured in DMEM medium and infected with VSV (MOI, 1). Simultaneously, cells were treated with either hnRNPA2B1 agonist (10 μM) or a negative control (physiological saline). Total RNA was collected from the cells at 24, 36, and 48 hours, and VSV mRNA was detected by real-time quantitative PCR.

[0268] VSV replication level such as Figure 10 As shown.

[0269] The results showed that the hnRNPA2B1 agonist significantly inhibited the replication of the RNA virus VSV. Furthermore, similar results were obtained using other compounds prepared in Example I in the same experiments.

[0270] Bioevaluation

[0271] The compounds disclosed herein are defined as hnRNPA2B1 agonists by: (i) binding to the hnRNPA2B1 protein, as demonstrated by Biacore assays of the high affinity of the compounds to the hnRNPA2B1 protein at concentrations as low as 20 μM; and (ii) demonstrated by cellular analysis that the compounds can broadly induce higher levels of IFN-β at concentrations as low as 20 μM.

[0272] In anti-cellular infection assays against different types of infectious agents, the hnRNPA2B1 agonist compounds of this disclosure significantly inhibited the replication of infectious agents, thereby exerting an anti-infective effect. Furthermore, since the hnRNPA2B1 agonist compounds of this disclosure can broadly induce higher levels of IFN-β, it is expected that these compounds will exert anti-infective effects through type I interferon action.

[0273] (i) Biacore T200 detection of the binding of the compound to hnRNPA2B1

[0274] The S-series CM5 chip was used; amino coupling reagents (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide EDC and N-hydroxysuccinimide NHS);

[0275] Coupling buffer: 10mM sodium acetate, pH 4.0, pH 4.5;

[0276] Run buffer: 10×PBS-P+;

[0277] Analytical grade DMSO, deionized water (0.22μm membrane filtration);

[0278] hnRNPA2B1 recombinant protein (concentration must be greater than 0.5 mg / mL, total protein amount must be at least 20 μg);

[0279] Compound: Mother liquor concentration greater than 20 mM, volume greater than 30 μL, purity greater than 90%, dissolved in 100% DMSO;

[0280] Other consumables: 96-well plates and sealing film, 1.5mL centrifuge tubes without caps, and type 2 rubber bottle caps.

[0281] Experimental steps:

[0282] 1. Place the running buffer 1×PBS and waste bottle in the designated locations, and then place the chip.

[0283] 2. Protein coupling. Select the immobilization program in the Wizard template, check flow cell 2 or 4, select amine as the method, select specify contact time and flow rate to achieve high coupling, and enter contact time 900s and flow rate 10μL / min.

[0284] 3. Prepare the required protein solution according to the instructions. Dilute the protein with sodium acetate at pH 4.0 to the required volume, with a final concentration of 20 μg / mL. Place 100 μL EDC, 100 μL NHS, and 140 μL ethanolamine into the corresponding positions.

[0285] 4. The system will automatically coat the chip surface with the target amount of hnRNPA2B1 protein and automatically generate a coupling report.

[0286] 5. Prepare the running buffer for interaction. For small molecule samples, use 1×PBS-P+ containing 5% DMSO: Dilute 105 mL of 10×PBS-P+ with deionized water to 1 L to prepare 1.05×PBS-P+.

[0287] Solvent-corrected stock solution containing 4.5% DMSO: 1.05×PBS-P + 9.5 mL + 0.45 mL DMSO

[0288] Solvent correction stock solution containing 5.8% DMSO: 1.05×PBS-P + 9.5mL + 0.58mL DMSO

[0289] Run buffer containing 5% DMSO: 1.05×PBS-P + 950mL + 50mL DMSO

[0290] Dilute the 10mM small molecule stock solution with DMSO-free 1.05×PBS-P+ buffer: 0.7μL small molecule stock solution + 6.3μL DMSO + 133μL 1.05×PBS-P+ buffer, for a total volume of 140μL and a final concentration of 50μM. Add to 96-well plates and seal with film.

[0291] After the test is completed, binding affinity is determined using Kinetics or Affinity mode.

[0292] (ii) Total RNA from cells can be extracted using TRIzol or a rapid RNA extraction kit.

[0293] 24-well plate cells (approximately 2-3 × 10⁻⁶) 5 Rapid RNA extraction (cells / well): Discard the culture medium, add 500 μL of RA2 lysis buffer to each well, lyse by pipetting several times, transfer to the inner sleeve of a dedicated adsorption tube, and centrifuge at 12000 rpm for 1 min at room temperature. Discard the filtrate, add 500 μL of wash buffer, and centrifuge at 12000 rpm for 1 min at room temperature. Repeat the washing process once. Discard the filtrate, replace the inner sleeve, and centrifuge at 12000 rpm for 2 min at room temperature without adding wash buffer. Transfer the inner sleeve to a new 1.5 mL centrifuge tube, add 25 μL of Elution Buffer to the center of the membrane, incubate at room temperature for 5 min, and centrifuge at 12000 rpm for 1 min at room temperature. The resulting filtrate is the RNA; determine the RNA concentration using Nanodrop One.

[0294] qRT-PCR: After determining the concentration of each sample, take 1 μg of total RNA and use ReverTra... The qRT-PCR RTMaster Mix reverse transcribes RNA into cDNA. The reverse transcription system and reaction conditions are as follows:

[0295] Reverse transcription reaction system (20 μL system)

[0296]

[0297] Reverse transcription reaction conditions (20 μL system)

[0298]

[0299] Add 60 μL of deionized water to the cDNA sample obtained by reverse transcription, mix well, and then centrifuge briefly. Primer sequence information used for qRT-PCR is shown in the table. The relative quantification of gene mRNA levels was performed using SYBR Green Realtime PCR Master Mix.

[0300] qRT-PCR reaction system (20 μL)

[0301]

[0302] qRT-PCR reaction conditions

[0303]

[0304] qRT-PCR primers

[0305]

[0306] All documents mentioned in this disclosure are incorporated herein by reference as if each document were individually incorporated herein by reference. Furthermore, it should be understood that after reading the foregoing teachings of this disclosure, those skilled in the art can make various alterations or modifications to this disclosure, and these equivalent forms also fall within the scope defined by the appended claims.

Claims

1. A product selected from: pharmaceutical compositions, pharmaceutical products, kits, or cassettes, comprising a fused cyclic compound of formula (I) or a pharmaceutically acceptable salt thereof, and a pharmaceutically or immunologically acceptable carrier or excipient. (I) in, L is -(CH2)n-, and n is 1, 2, or 3; X is a halogen; R 1 For -NR 4 R 5 , where R 4 R 5 Each independently is H or C 1-6 alkyl; R 2 and R 3 Each independently is C 1-3 Alkyl, the C 1-3 The alkyl group is either unsubstituted or substituted with a 4-6 membered heterocyclic alkyl group containing one nitrogen atom; Or R 2 and R 3 Together with the N atoms to which they are attached, they form a piperazine group, which is bonded by a 4-6 membered heterocyclic alkyl group. 1-6 Alkyl or hydroxyl C 1-6 Alkyl substitution, wherein 1, 2, 3 or 4 ring atoms in the 4-6 membered heterocyclic alkyl group are heteroatoms independently selected from O, S and N, and the remainder are carbon atoms; The product is used to induce type I interferon production in subjects and / or to prevent and / or treat infectious diseases and / or infection-related diseases and / or symptoms in subjects; The product also contains one or more other active substances for the prevention or treatment of infectious diseases and their related symptoms and / or signs.

2. The product of claim 1, wherein the other active substance is one or more selected from the group consisting of: commonly used clinical antibiotics selected from β-lactams, aminoglycosides, tetracyclines, chloramphenicols, macrolides, antifungal antibiotics, and antituberculosis antibiotics; commonly used clinical antiviral drugs selected from tricyclic amines, pyrophosphates, protease inhibitors, nucleoside analogs, interferons, and antisense oligonucleotides; and commonly used clinical immunosuppressants selected from glucocorticoids, cyclophosphamide, chloroquine, cyclosporine A, traditional Chinese medicine preparations, and anti-TNF monoclonal antibodies.

3. The product as described in claim 2, wherein the β-lactam is selected from penicillins and cephalosporins, and the traditional Chinese medicine preparation is selected from Tripterygium wilfordii.

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

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

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

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

8. The product of claim 7, wherein R c It is a hydroxyl group or a 4-6 membered heterocyclic alkyl group containing one nitrogen atom.

9. The product of claim 7, wherein R c It is hydroxyl or pyrroleyl.

10. The product of claim 7, wherein R c It is hydroxyl or 1-pyrrolidinyl.

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

12. The use of the fused cyclic compound of formula (I) or a pharmaceutically acceptable salt thereof in the preparation of products for inducing type I interferon production in subjects or for the prevention and / or treatment of infectious diseases and / or infection-related diseases and / or symptoms, wherein said products are selected from: pharmaceutical compositions, pharmaceutical products, kits or cassettes. (I) in, L is -(CH2)n-, and n is 1, 2, or 3; X is a halogen; R 1 For -NR 4 R 5 , where R 4 R 5 Each independently is H or C 1-6 alkyl; R 2 and R 3 Each independently is C 1-3 Alkyl, the C 1-3 The alkyl group is either unsubstituted or substituted with a 4-6 membered heterocyclic alkyl group containing one nitrogen atom; Or R 2 and R 3 Together with the N atoms to which they are attached, they form a piperazine group, which is bonded by a 4-6 membered heterocyclic alkyl group. 1-6 Alkyl or hydroxy C 1-6 Alkyl substitution, wherein 1, 2, 3 or 4 ring atoms in the 4-6 membered heterocyclic alkyl group are heteroatoms independently selected from O, S and N, and the remainder are carbon atoms.

13. The application as described in claim 12, wherein, The object is a human or a non-human mammal; and / or Wherein, the production of type I interferon is mediated by hnRNPA2B1; and / or The infectious disease in the subject can benefit from the production or increased levels of type I interferon in the body; and / or Wherein, the infectious disease is caused by chemical, physical, or biological infection; and / or Wherein, the infection-related diseases and / or symptoms are selected from one or more of the following groups: pathological damage caused by infection; insufficient or excessive production of cytokines after infection; endotoxic shock or death; inflammatory damage to organs; multiple organ failure; chronic inflammatory diseases caused by infection; and / or The product also contains one or more other active substances for the prevention or treatment of infectious diseases and their related symptoms and / or signs.

14. The application as described in claim 13, wherein the object is a gorilla, chimpanzee, pet cat, dog, guinea pig, rabbit, cow, horse, donkey, mule, camel, pig, sheep, chicken, duck, goose, deer, mink, otter, or musk deer; and / or The infectious disease is caused by a virus, bacteria, parasite, and / or fungus; and / or The infection-related disease and / or symptoms are selected from one or more of the following groups: insufficient or excessive interferon production; multiple organ failure, wherein the organs are selected from: liver, spleen, brain, kidney, heart, lung, stomach, intestine; autoimmune diseases; and / or The other active substances are selected from one or more of the following groups: commonly used clinical antibiotics selected from β-lactams, aminoglycosides, tetracyclines, chloramphenicols, macrolides, antifungal antibiotics, and antituberculosis antibiotics; commonly used clinical antiviral drugs selected from tricyclic amines, pyrophosphates, protease inhibitors, nucleoside analogs, interferons, and antisense oligonucleotides; and commonly used clinical immunosuppressants selected from glucocorticoids, cyclophosphamide, chloroquine, cyclosporine A, traditional Chinese medicine preparations, and anti-TNF monoclonal antibodies.

15. The application of claim 14, wherein the infectious disease is caused by a DNA viral infection; and / or The infection-related disease and / or symptoms are selected from one or more of the following groups: inflammatory bowel disease, rheumatoid arthritis, systemic lupus erythematosus, chronic nephritis, tuberculosis, and chronic gastrointestinal diseases.

16. The application of claim 15, wherein the infectious disease is an infection caused by one or more viruses selected from the group consisting of: herpes simplex virus, hepatitis B virus, adenovirus, poxvirus, microvirus, adeno-associated virus; and / or The β-lactams are selected from penicillins and cephalosporins, and the traditional Chinese medicine preparation is selected from Tripterygium wilfordii.

17. The application as described in claim 12, in, X is either F or Cl; L is -(CH2)n-, where n is 1 or 2; R 4 R 5 Each can be H, methyl, or ethyl.

18. The application as described in claim 12, in, X is F; L is -(CH2)n-, where n is 2; R 4 R 5 Each can be H, methyl, or ethyl.

19. The application as described in claim 12, in, X is F; L is -(CH2)n-, where n is 2; R 4 R 5 Each can be independently H, methyl, or ethyl; R 2 and R 3 Each is independently methyl, ethyl, or isopropyl, wherein the methyl, ethyl, or isopropyl groups are either unsubstituted or substituted with pyrrolidinyl groups; Or R 2 and R 3 Together with the N atoms to which they are attached, they form a piperazine group, which is substituted with a pyrrolidinyl ethyl or a hydroxyethyl.

20. The application as described in claim 12, wherein the compound has the following formula (II): (II) in, R a R b Each independently is H or C 1-6 alkyl; M is -(CH2)n-, where n is 1, 2 or 3; X is either F or Cl; R c It is a hydroxyl group containing 1-2 heteroatoms selected from nitrogen, oxygen or sulfur, and is a 4-6 membered heterocyclic alkyl group.

21. The application as described in claim 20, wherein R c It is a hydroxyl group or a 4-6 membered heterocyclic alkyl group containing one nitrogen atom.

22. The application as described in claim 20, wherein R c It is hydroxyl or pyrroleyl.

23. The application as described in claim 20, wherein R c It is hydroxyl or 1-pyrrolidinyl.

24. The application as described in claim 12, wherein the compound is selected from the group consisting of: , , , , ,or , or its hydrochloride salt.

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