Inhibitors of SLC15A4 small molecules with anti-inflammatory activity

Abstract

Small molecule inhibitors of SLC15A4 and methods for using them to treat pDC-mediated diseases and conditions have been disclosed.

Description

[0001] Cross-reference to related applications

[0002] This application claims priority to U.S. Provisional Patent Application No. 62 / 981,907, filed on February 26, 2020, which is incorporated herein by reference in its entirety. Technical Field

[0003] This disclosure relates to novel chemical compounds and methods for inhibiting SLC15A4. Background Technology

[0004] Recognition of infectious pathogens relies on a series of germline-encoded immune sensors called model receptors (TLRs) and NOD-like receptors (NLRs). TLRs are membrane-sensitive sensors that scan for microbial PAMPs in the extracellular environment, while NLRs monitor the cytoplasmic environment. Viral and bacterial nucleic acids are important PAMPs recognized by several TLRs, including TLR3, TLR7, TLR8, and TLR9. Ligand binding to these sensors leads to signaling events that induce the expression of several genes that enhance the immune response to pathogen killing, including inflammatory cytokines, stimulating immune cytokines, chemokines, and co-stimulatory molecules. 1,2 However, improper recognition of host nucleic acids can lead to autoimmune diseases or autoinflammatory conditions. 3-6 Autoimmunity arises through several concurrent mechanisms associated with the presence of self-reactive immune cell subsets and the loss of immune tolerance. Loss of tolerance during central and peripheral differentiation of the adaptive immune response can lead to uncontrolled activation of self-reactive B and T cells, inducing autoimmunity aided by innate immune cells. TLR signaling plays a crucial role in the activation of the adaptive immune system by inducing the production of pro-inflammatory cytokines, and persistent activation or dysregulation of TLR signaling directly contributes to the pathogenesis of autoimmunity. 7 An important finding is that activation of endolysosomal nucleic acid-sensing TLRs and the production of type I interferon (IFN-I) (particularly through APC-like plasmacytoid dendritic cells (pDCs)) are central drivers of pathogenic events. 8 .

[0005] pDCs are a specialized subset of recirculating dendritic cells that act as early sentinels in pathogen surveillance. They respond to TLR7 and TLR9 (TLR7 / 9) receptors on microbial nucleic acids and endogenous nucleic acids. 9,10Upon recognition, pDCs produce approximately 1000 times more type 1 IFN (IFN-I) than any other cell type. TLR7 / 9 activation in pDCs can also induce other cytokines (IL-12, IL-6, TNFα) and inflammatory chemokines. 11 Evidence suggests that pDCs also activate B cells, act as APCs, and promote immune regulation and tolerance. 12-15 Given their central role in inflammation, it is perhaps not surprising that pDCs are causal effectors in the pathogenesis of many autoimmune diseases, including lupus and psoriasis. One of the closest links between pDCs and autoimmune diseases is during systemic autoimmune disease (SLE). 8 In most mouse models, lupus is IFN-I dependent, and gene deletion or neutralization of IFN-I signaling can prevent or improve the disease. 7 In addition, approximately 70% of SLE patients exhibit elevated IFN-I levels. 16 Furthermore, clinical trials using IFN-I receptor neutralizing antibodies are currently underway and have yielded promising outcomes in clinical lupus patients. pDCs have also been detected in the cerebrospinal fluid of patients with multiple sclerosis (MS). 17 Furthermore, it accumulates in demyelinating lesions of inflamed MS brains. 18 .

[0006] Small-molecule immunomodulatory drugs have been developed to control harmful immune responses during inflammation, transplantation, and autoimmune diseases. Corticosteroids, calmodulin inhibitors, nonsteroidal anti-inflammatory drugs (NSAIDs), mTOR inhibitors, and kinase inhibitors have been used to treat autoimmune diseases and improve transplant efficacy. Recently, agents such as Gilenya and Tecfidera have shown efficacy in alleviating disease progression and symptoms in patients with multiple sclerosis. However, most of these treatments are general inflammatory modulators or broadly applicable immunosuppressants, and they can participate in signaling pathways common to almost all cell types, leading to undesirable side effects and limiting their use. 19 Monoclonal antibody (mAb) therapies targeting specific immune targets (such as checkpoint inhibitors (anti-CTLA-4) and anti-TNF) have been developed, with some success; however, they are generally only effective in a subset of patients and serious immune-related adverse events (irAEs) are frequently observed. 19,20 Currently, there are no available therapeutic agents targeting pDC-mediated IFN-I (a major driver of many autoimmune diseases).21 Therefore, there is currently an unmet need for novel small molecule therapeutics for sanguine pDC-mediated diseases.

[0007] SLC15A4 plays a central role in pDC-mediated inflammation and autoimmunity. SLC15A4 (also known as Proton / Histidine Transporter 1, PHT1) is a 12-membered transmembrane protein whose gene expression is primarily confined to APCs (especially pDCs and B cells). 22-24 SLC15A4 is a member of the SLC15 family, which includes the proton / histidine transporter SLC15A3 (PHT2) and the dipeptide / tripeptide transporters SLC15A1 (PEPT1) and SLC15A2 (PEPT2). Both SLC15A3 and SLC15A4 contain an acidic bis-leucine motif that mediates endosome / lysosome localization and are labeled as dipeptide or tripeptide cotransporters. 25 The lysosomes and endosomes are acidic, indicating that SLC15A3 and SLC15A4 share 60% sequence identity and utilize an outward-oriented proton gradient to transport short peptides into the cytosol. 25,26 However, the substrates for lysosomal SLC15A3 and SLC15A4 are not yet fully understood. Several studies have shown that SLC15A4 can transport bacterial-derived peptidoglycans (such as MDP and Tri-DAP, which are ligands for the immunosensitive substances NOD1 and NOD2), thereby leading to their activation. 27-30 SLC15A4 is also closely related to TLR7 / 9-mediated signaling and IFN-I production. Specifically, studies have shown that these two factors lead to pDCs in Slc15a4 loss-of-function mutants (called "feeble") and Slc15a4 knockout mice that exhibit defective IFN-I and TNF-α, IL-6, and IL-12 production after TLR stimulation, but show normal development in other aspects. 31,29,30,32 This deficiency is not due to impaired TLR ligand uptake or IFN-I secretion, and affects both TLR7 and TLR9 signaling pathways. Crucially, SLc15a4 nullipopulation mice showed a significant reduction in clinical lupus manifestations and prolonged lifespan. 32Although SLC15A3 and SLC15A4 are thought to have similar functions, the disease-mitigating effects of SLC15A4 mutations and deletions suggest significant functional differences between these two transporters, or that the expression of either is essential for optimal function. Furthermore, genome-wide association studies (GWAS) have shown that SLC15A4 (but not SLC15A3) is closely associated with inflammatory diseases such as systemic lupus erythematosus (SLE) and inflammatory bowel disease (IBD). 33,34 However, the exact mechanisms by which SLC15A4 leads to these processes remain to be determined. Nevertheless, the fundamental pathogenic role of the pDC / TLR / IFN-I axis and its role in improving autoimmune diseases in SLC15A4 loss-of-function mouse models identify SLC15A4 as a key mediator of inflammation and provides a strong basis for the characterization of SLC15A4 and the development of inhibitors.

[0008] Biological and chemical probe discoveries of SLC. The solute-like carrier (SLC) protein family is the largest group of membrane transport proteins, containing 456 members distributed across 52 subfamilies. SLC is not only associated with many diseases caused by genetic polymorphisms, but also plays a role in tumorigenesis, autoimmune diseases, and metabolic disorders. 35-37 While important, most (>30%) of SLCs have few or no properties, and the vast majority (>80%) lack chemical probes. 38 One of the most significant obstacles is their complex overall membrane morphology, requiring intact membranes to retain native functional properties. The difficulties in expressing and purifying native SLCs limit their use to: traditional high-throughput screening (HTS) methods, and commonly used in vitro biochemical studies for annotating substrate ranges, measuring transport rates, and examining the effects of various interfering factors (e.g., mutations, inhibitors) on transport. 38 Due to inherent technical challenges, only about 10 human SLCs with a structure exist, along with a small number of SLCs existing in multiple conformations or bound to substrates or drugs. This limits any possibilities for structure-based drug design. 39 Cell-based and animal-based models for SLC research can be similarly challenging because genetic perturbations can become complex through overlapping specificities, compensatory mechanisms, and toxicities, limiting research to a subpopulation of SLCs and sometimes masking the relative contribution of transporters to the function or phenotype under investigation.35,36,40,41 Given these challenges, there is an urgent need for new methods to study SLC biology and to develop usable SLC-targeting chemical probes.

[0009] Previous studies have identified SLC15A4 as playing a unique and crucial role in the production of IFN-I and other inflammatory cytokines in pDCs and in the pathogenesis of autoimmune diseases, thus elevating SLC15A4 to a potential therapeutic target for such conditions. However, to date, SLC15A4 remains untreatable and no inhibitors have been publicly disclosed. This application not only describes a feasible chemical proteomics strategy to deconvolve the mechanisms by which SLC15A4 controls TLR signaling but also evaluates the therapeutic potential of SLC15A4 for treating pDC-mediated conditions.

[0010] There are no clinically approved drugs that specifically target pDCs and their IFN-I production, as well as nucleotide-binding TLR signaling, which are central factors in the pathogenesis of many autoimmune diseases, such as lupus, Crohn's disease, irritable bowl syndrome (IBS), type 1 diabetes, psoriasis, and possibly even MS. Crucially, SLC15A4 is primarily expressed in antigen-presenting cells (particularly pDCs, B cells, and macrophages) that directly contribute to the pathogenesis of autoimmune diseases, making SLC15A4 a highly relevant therapeutic target for developing compounds that selectively suppress inflammation. Summary of the Invention

[0011] The applicant has discovered novel SLC154A inhibitor compounds and evaluated the performance and utility of some representative examples of such compounds, both for biochemical efficacy (e.g., evaluating % IFNα inhibition ± SD and % transport inhibition in human pDC).

[0012] In several embodiments, this disclosure relates to compounds of formula (I) or (II) or pharmaceutically acceptable salts thereof:

[0013] ,

[0014] in

[0015] X is NR 4 Or S;

[0016] R 1 It is H or -C(O)-C 1-10 alkyl;

[0017] R 2 It is a heterocyclic or aryl group;

[0018] R 3 It is a halogen, -CHF2 or -CF3;

[0019] R 4 It is H, -C 1-10 Alkyl, -C(O)-C 1-10 Alkyl, -C(O)-C 3-10 Cycloalkyl, -S(O)2-C 1-10 alkyl or

[0020] ;as well as

[0021] n is 0, 1, 2, 3 or 4.

[0022] In several implementation schemes, this disclosure relates to Figure 15 Compounds.

[0023] In several embodiments, this disclosure relates to methods for treating pDC, B cell, macrophage, or monocyte-mediated conditions. Brief description of the attached diagram

[0025] Figure 1 A through 1C illustrate fragment-based ligandability mapping in cells (FbLMiC). An overview of the methods and specialized chemical libraries is provided. (A) Fully functionalized fragment (FFF) probes consist of drug-like fragments and retrieval tags that enable covalent capture of fragment-bound protein targets directly in cells after UV irradiation. Fragment targets and fragment interaction sites are identified and quantified using mass spectrometry-based and gel-based methods. (B) The general structure of the FFF library shows constant affinity tag regions (red) containing photoreactive groups (diazidine) and potential affinity groups (alkynes), and variable regions (blue) containing fragment recognition elements that bind to proteins. Some examples of fragments are shown. (C) A subset of proteins coordinating with FFF ligands provides first evidence for druggability (non-drugbank) and functional classification of identified druggable proteins (drugbank). (46,47)

[0026] Figures 2A to 2DThe development of chemical proteomics for SLC chemical probes is shown. (A) An overview of SLC interactions identified using FbLMiC in HEK293T, K562, and human PBMC cells. SLCs are considered FFF targets if they exhibit >5-fold reproducibility enrichment relative to control methylFFF probes (at least in biological replication) and show chemotype selectivity (e.g., not enriched by all FFFs). The X-axis shows SLC subfamilies, and the Y-axis shows fragments of SLC subfamilies considered to be FFF targets. (B) Structures of FFF3, a fragment-based inhibitor (CP22), and a control compound (CP26) used in a previously published (46) functional study of SLC25A20. (C) FFF3 probe labeling sites mapped onto the homologous SLC25A20 structure (brown). Exemplary MS1 chromatograms of probe-labeled trypsin peptides are shown in blue. (D) CP22 increases the levels of long-chain ester carnitine in HSC5 cells. Data = Mean ± SD; For the treatment group, ** p < 0.01 *** p < 0.001 and **** p < 0.0001; n = 3 to 5.

[0027] Figures 3A to 3D The development of chemical proteomics for SLC15A4 chemical probes is shown. (A) Structures of FFF probes identified as involved in SLC15A4 in proteomics experiments (see full text of the experimental description). The ability of all probes to inhibit IFN-I production in human pDCs was examined, with 5 showing the highest activity. No structurally similar probe 6 was found to be involved in SLC15A4 or inhibit IFN-I production. (B) Gel-based competitive readouts show the interaction of excess 5-comp competing with 5 in human PBMCs. (C) Isotope reduction demethylation thermograms show the protein enriched by excess 5-comp or 6-comp (80 mM) competing with 5 (20 mM) in human PBMCs. The inset shows the identities of the top 15 competing targets. (D) Example MS1 of SLC15A4 trypsin peptide from the competitive experiment.

[0028] Figures 4A to 4D The SLC15A4 chemical probe inhibits inflammatory cytokines (IFN-I and IL-6) in primary mouse and human pDCs. Inhibition of IFN-I production in isolated human pDCs (A and B) and mouse pDCs (C). (D) Inhibition of IL-6 in primary mouse pDCs. Mean ± SD (n = 3).

[0029] Figures 5A to 5CThe progress of the SLC15A4 transporter reporter gene assay is shown. (A) Fluorescence micrograph of A549 cells stably transfected with SLC15A4-mCherryWT (top) and mutants (L14A, L15A, L318A, V319A, bottom). SLC15A4 mutant expression is localized to the cell membrane. (B) Schematic diagram of the SLC15A4 NFkB transporter assay run in 96-well format. (C) Luciferase signaling produced by SLC15A4 mutants instead of WT after treatment with MDP or Tri-DAP. Luciferase expression was inhibited in the presence of triptolide (an NFkB inhibitor) and 5 (instead of 6). Mean ± SD (n = 3).

[0030] Figures 6A to 6E SAR studies and functional evaluations of the SLC15A4 inhibitors are shown. (A) Structure of 5-comp and two common synthetic routes used for SAR studies. (B) Structure of 5-comp analogs. (C) Representative cytotoxicity profiles of isolated human pDCs treated with 10 mM of each compound after 24 hours. Readouts were performed using Cell Titer Glo. (D) Correlation plot of IFN-I inhibition (x-axis) versus transport inhibition (y-axis) at 10 mM for each compound. (E) Dose-dependent inhibition in primary human pDCs in the case of lead analog 8. Mean ± SD (n = 3).

[0031] Figure 7 This is a table summarizing IFNα inhibition and transport inhibition in human pDC.

[0032] Figure 8 This is the IC50 table for SLC15A4 inhibitors.

[0033] Figure 9 This is a series of graphs showing how SLC15A4 inhibitors block MDP transport in a dose-dependent manner, where AJ2-3A and AJ2-30 are active controls, while AJ2-18 and AJ2-22 are inactive controls.

[0034] Figure 10 Compounds involved in blocking endogenous NOD signaling in THP cells by SLC15A4 are shown.

[0035] Figure 11 Compounds involved in blocking endogenous NOD signaling in human and mouse macrophages via SLC15A4 are shown, with AJ2-30 being the active control and AJ2-18 being the inactive control.

[0036] Figure 12 This demonstrates that the SLC15A4 inhibitor inhibits TLR9-mediated B cell activation.

[0037] Figure 13 The results showed that the SLC15A4 inhibitors were inactive in immune cells from SLC15A4-ineffective mice, with AJ2-3A and AJ2-30 being active controls and AJ2-18 and AJ2-22 being inactive controls.

[0038] Figure 14 The in vivo efficacy of the SLC15A4 inhibitor in a simple inflammation model was demonstrated. The compound (or carrier) was co-injected with CpG (TLR9) into mice; serum was collected 6 hours later to measure cytokines (single dose); AJ2-3 and AJ2-30 were active controls, while AJ2-22 was an inactive control.

[0039] Figure 15 It is the structure of the SLC15A4 inhibitors AJ2-1 to AJ2-92 and AJ2-CP53.

[0040] Detailed description

[0041] In several embodiments, this disclosure relates to compounds that inhibit SLC15A4. In several embodiments, the compounds are selective for SLC15A4.

[0042] The compound can be used to treat pDC, B cell, macrophage or monocyte-mediated conditions.

[0043] definition

[0044] For convenience, certain terms used in the specification, embodiments, and appended claims are summarized herein before further description of this disclosure. These definitions should be interpreted in light of the remainder of this disclosure and understood by those skilled in the art. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art.

[0045] To make this disclosure easier to understand, certain terms and phrases are defined below and throughout the specification.

[0046] Nouns without quantifiers are used in this text to refer to one or more (i.e., at least one). For example, "element" means one or more elements.

[0047] The phrase “and / or” as used herein in the specification and claims should be understood to mean “any one or both” of the elements so combined (i.e., elements present in some cases but not in others). Multiple elements listed with “and / or” should be interpreted in the same way, i.e., “one or more” of the elements so combined. Other elements may optionally be present in addition to those specifically identified by the “and / or” clause, whether related to or unrelated to those specifically identified. Thus, as a non-limiting example, when used in conjunction with open-ended language such as “comprising / including,” references to “A and / or B” may: in one embodiment refer only to A (optionally including elements other than B); or in another embodiment refer only to B (optionally including elements other than A); or in yet another embodiment refer to both A and B (optionally including other elements); and so on.

[0048] As used herein in the specification and claims, “or / or” should be understood to have the same meaning as “and / or” as defined above. For example, when separating items in a list, “or / or” or “and / or” should be interpreted as inclusive, i.e., including multiple elements or at least one of the elements in the list, but also including more than one, as well as optional additional items not listed. Only when the opposite term is explicitly indicated, such as “only one of…” or “exact one of…”, or when used in the claims, “consisting of…”, will refer to the inclusion of multiple elements or exactly one of the elements in the list. In general, the term “or / or” as used herein, when preceded by an exclusive term such as “any,” “one of…,” “only one of…,” or “exact one of…,” should only be interpreted as indicating an alternative to exclusivity (i.e., “one or the other, but not both”). “Constitutes mainly of…” when used in the claims should have its ordinary meaning as used in the field of patent law.

[0049] As used herein in the specification and claims, the phrase “at least one” in relation to a list of one or more elements should be understood to mean at least one element selected from any one or more elements in the list of elements, but does not necessarily include at least one of each element specifically listed in the list of elements, nor exclude any combination of elements in the list of elements. This definition also allows for the optional presence of elements other than those specifically identified in the list of elements referred to by the phrase “at least one,” whether related to or unrelated to those specifically identified elements. Thus, as a non-limiting example, “at least one of A and B” (or, equivalently, “at least one of A or B,” or equivalently, “at least one of A and / or B”) could mean: in one embodiment, at least one, optionally including more than one A, and no B (and optionally including elements other than B); or in another embodiment, at least one, optionally including more than one B, and no A (and optionally including elements other than A); or in yet another embodiment, at least one, optionally including more than one A, and at least one, optionally including more than one B (and optionally including other elements); etc.

[0050] It should also be understood that, unless explicitly stated otherwise, in any method claimed herein that includes more than one step or action, the order of the steps or actions of the method is not necessarily limited to the order in which the steps or actions of the method are recorded.

[0051] In the claims and in the foregoing description, all transitional phrases such as “comprising,” “including,” “carrying,” “having,” “containing,” “involving,” “holding,” “constituting,” etc., shall be understood to be open-ended, i.e., intended to include, but not limited to. Only the transitional phrases “constituting of,” and “constituting substantially of,” shall be closed or semi-closed transitional phrases, respectively, as described in Section 2111.03 of the U.S. Patent and Trademark Office Patent Examination Procedure Manual.

[0052] The various compounds included in the compositions of this disclosure may exist in specific geometric or stereoisomeric forms. Additionally, the polymers of this disclosure may also be optically active. This disclosure contemplates all such compounds, including cis and trans isomers, R- and S-enantiomers, diastereomers, (d)-isomers, (l)-isomers, racemic mixtures thereof, and other mixtures thereof, all of which fall within the scope of this disclosure. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers, and mixtures thereof, are intended to be included in this disclosure.

[0053] For example, if a particular enantiomer of the compound of this disclosure is desired, it can be prepared by asymmetric synthesis or by derivatization with a chiral auxiliary, wherein the resulting diastereomeric mixture is isolated and the auxiliary group is cleaved to provide a pure desired enantiomer. Alternatively, when the molecule contains a basic functional group (e.g., amino) or an acidic functional group (e.g., carboxyl), a diastereomeric salt is formed with a suitable optically active acid or base, followed by decomposition of the resulting diastereomeric mixture by fractional crystallization or chromatographic means known in the art, and then the pure enantiomer is recovered.

[0054] The structures described herein are also intended to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, by replacing hydrogen with deuterium or tritium, or by using... 13 C- or 14 Compounds produced by C-enrichment of carbon substitutes for carbon are within the scope of this disclosure.

[0055] As used herein, the term "prodrug" encompasses compounds that are converted into therapeutically active agents under physiological conditions. A common method for preparing prodrugs involves selecting a moiety that is hydrolyzed under physiological conditions to reveal the desired molecule. In other embodiments, the prodrug is converted by the enzymatic activity of a host animal.

[0056] As used herein, the phrase “medicinal excipient” or “medicinal carrier” means a pharmaceutically usable substance, composition, or carrier, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating substance, relating to the carrying or transport of a subject chemical substance from one organ or site of the body to another organ or site of the body. Each carrier must be “usable” in the sense of being compatible with other components of the formulation, harmless to the patient, and substantially nonpyrogenic. Some examples of substances that can be used as pharmaceutical carriers include: (1) sugars, such as lactose, glucose, and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate; (4) tragacanth powder; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and so on. Soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerol, sorbitol, mannitol, and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffers, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotropic saline; (18) Ringer's solution; (19) ethanol; (20) phosphate buffer solution; and (21) other non-toxic and compatible substances used in pharmaceutical formulations. In several embodiments, the pharmaceutical compositions of this disclosure are pyrogen-free, i.e., they do not induce a significant increase in temperature when administered to a patient.

[0057] The term "pharmaceutical salt" refers to the relatively non-toxic inorganic and organic acid addition salt of a compound. These salts can be prepared in situ during the final separation and purification of the compound, or by reacting the purified compound, in its free base form, with a suitable organic or inorganic acid and then separating the resulting salt. Some representative salts include hydrobromide, hydrochloride, sulfate, hydrogen sulfate, phosphate, nitrate, acetate, valerate, oleate, palmitate, stearate, laurate, benzoate, lactate, phosphate, toluenesulfonate, citrate, maleate, fumarate, succinate, tartrate, naphthalate, methanesulfonate, gluconate, lactobionate, and laurylsulfonate, etc. (see, for example, Berge et al. (1977) "Pharmaceutical Salts", J. Pharm. Sci. 66:1-19.).

[0058] In other cases, compounds that can be used in the methods of this disclosure may contain one or more acidic functional groups and are therefore capable of forming pharmaceutically acceptable salts with pharmaceutically acceptable bases. In these cases, the term "pharmaceutically acceptable salt" refers to a relatively non-toxic inorganic or organic base addition salt of the compound. These salts can also be prepared in situ during the final separation and purification of the compound, or by reacting the purified compound in its free acid form with a suitable base (e.g., a hydroxide, carbonate, or bicarbonate of a pharmaceutically acceptable metal cation), with ammonia, or alone with a pharmaceutically acceptable primary, secondary, or tertiary organic amine. Some representative bases or alkaline earth salts include lithium, sodium, potassium, calcium, magnesium, and aluminum salts. Some representative organic amines that can be used to form base addition salts include ethylamine, diethylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine, etc. (see, for example, Berge et al., ibid.).

[0059] For therapeutic purposes, a “therapeutic effective amount” (or “effective amount”) of a compound refers to the amount of the compound in a formulation that, when administered as part of a desired dosing regimen (in mammals, such as humans), reduces symptoms, improves conditions, or slows the onset of disease symptoms, based on clinically acceptable criteria for the treatment of the disorder or condition or for cosmetic purposes, for example, with a reasonable benefit / risk ratio applicable to any medical treatment.

[0060] The term "preventive or therapeutic" treatment is recognized in the art and includes the administration of one or more subject compounds to a host. A treatment is preventive (i.e., it protects the host from developing an undesirable condition) if it is administered before the clinical manifestation of an undesirable symptom (e.g., a disease or other undesirable state in the host animal), and therapeutic (i.e., it aims to reduce, alleviate, or stabilize an existing undesirable symptom or its side effects) if it is administered after the manifestation of an undesirable symptom.

[0061] The terms "patient" or "object" refer to a mammal requiring specific treatment. In many embodiments, the patient or object is a primate, canine, feline, or equine. In many embodiments, the patient or object is a human.

[0062] Aliphatic chains include the categories of alkyl, alkenyl, and alkynyl groups as defined below. Straight aliphatic chains are limited to the unbranched carbon chain portion. As used herein, the term "aliphatic group" refers to a straight-chain, branched, or cyclic aliphatic hydrocarbon group and includes saturated and unsaturated aliphatic groups such as alkyl, alkenyl, or alkynyl groups.

[0063] "Alkyl" refers to a fully saturated cyclic or acyclic, branched or unbranched carbon chain moiety having a specified number of carbon atoms, or up to 30 carbon atoms if not specified. For example, alkyl groups with 1 to 8 carbon atoms refer to moieties such as methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, and octyl, as well as those moieties that are positional isomers of these moieties. Alkyl groups with 10 to 30 carbon atoms include decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecanyl, octadecyl, nonadecanyl, eicosyl, dodecyl, tridecyl, and tetradecyl. In many embodiments, straight-chain or branched alkyl groups have 30 or fewer carbon atoms in their main chain (e.g., for straight chains of C1 to C2). 30 For branches C3 to C 30 (1), or 20 or fewer. Alkyl groups may be substituted or unsubstituted.

[0064] As used herein, the term "alkylene" refers to an alkyl group having a specific number of carbon atoms (e.g., 2 to 12 carbon atoms) comprising two connection points on its longest carbon chain to the remainder of the compound. Some non-limiting examples of alkylenes include methylene-(CH2)-, ethylene-(CH2CH2)-, n-propylene-(CH2CH2CH2)-, isopropylene-(CH2CH(CH3))-, etc. Alkylenes can be cyclic or acyclic, branched or unbranched carbon chain portions, and may optionally be substituted with one or more substituents.

[0065] "Cycloalkyl" refers to a saturated carbon ring, whether monocyclic, bicyclic, bridged, spirocyclic, or polycyclic, each having 3 to 12 carbon atoms. Similarly, some cycloalkyl groups have 3 to 10 carbon atoms in their ring structure, while others have 3 to 6 carbon atoms. Cycloalkyl groups can be substituted or unsubstituted.

[0066] Unless otherwise specified, “lower alkyl” as used herein means an alkyl group as defined above but having 1 to 10 carbon atoms or 1 to 6 carbon atoms in its main chain structure, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, and tert-butyl. Similarly, “lower alkenyl” and “lower alkynyl” have similar chain lengths. Throughout the application, the alkyl group may be a lower alkyl group. In several embodiments, the substituents designated herein as alkyl groups are lower alkyl groups.

[0067] "Alkenyl" refers to any cyclic or acyclic, branched or unbranched unsaturated carbon chain portion having a specified number of carbon atoms, or up to 26 carbon atoms if there is no restriction on the specified number of carbon atoms; and having one or more double bonds in the portion. Examples of alkenyl groups with 6 to 26 carbon atoms include hexenyl, heptenyl, octenyl, nonenyl, decenyl, undecenyl, dodecenyl, tridecenyl, tetradecenyl, pentadecenyl, hexadecenyl, heptadecenyl, octadecenyl, nonadecenyl, icosyl, icosyl, icosyl, tridecenyl, and icosyl-tetradecenyl, as well as their various isomers, wherein the unsaturated bond may be located anywhere in the portion and may have (Z) or (E) configurations with respect to the double bond.

[0068] "Alkyne" refers to a hydrocarbon group within the alkenyl group range, but with one or more triple bonds in that group.

[0069] The term "alkylthio" refers to an alkyl group as defined above, having a sulfur moiety attached thereto. In various embodiments, the "alkylthio" moiety is composed of -(S)-alkyl, -(S)-alkenyl, -(S)-ynyl, and -(S)-(CH2). m -R 1 One representation in which m and R 1 The following definition applies. Representative alkyl thio groups include methyl thio, ethyl thio, etc. As used herein, the term "alkoxy" refers to an alkyl group having an oxygen moiety attached to it. Representative alkoxy groups include methoxy, ethoxy, propoxy, tert-butoxy, etc. An "ether" is two hydrocarbons covalently linked by oxygen. Therefore, the substituent of the alkyl group that makes it an ether is or similar to an alkoxy group, for example, it may be -O-alkyl, -O-alkenyl, -O-ynyl, O-(CH2) m -R 10 One representation in which m and R 10 As described below.

[0070] The terms "amine" and "amino" are recognized in the art and refer to both unsubstituted and substituted amines, for example, portions that can be represented by the following formula:

[0071]

[0072] Where R 11 R 12 and R 13 Each can independently represent hydrogen, alkyl, alkenyl, or -(CH2). m -R 10 , or R 11 and R 12 Together with the N atoms they are attached to, they form heterocycles with 4 to 8 atoms in the ring structure; R 10 Represents an alkenyl, aryl, cycloalkyl, cycloalkenyl, heterocyclic, or polycyclic group; and m is an integer from 0 to 8. In several embodiments, R 11 or R 12 Only one of them can be a carbonyl group, such as R. 11 R 12 It does not form an imide when combined with nitrogen. In even more different embodiments, R 11 and R 12 (and optional R) 13 Each of these can independently represent hydrogen, alkyl, alkenyl, or -(CH2). m -R 10 Therefore, the term "alkylamine" as used herein means an amino group as defined above, having a substituted or unsubstituted alkyl group attached to it, i.e., R 11 and R 12 At least one of them is an alkyl group. In several embodiments, the amino or alkylamine is basic, which means that it is a conjugate acid with a pKa > 7.00, i.e., the protonated form of these functional groups has a pKa greater than about 7.00 relative to water.

[0073] As used herein, the term "amide" refers to the following groups:

[0074]

[0075] Each R 14 Independently representing a hydrogen or hydrocarbon group, or two Rs 14 Together with the N atoms they are attached to, they form heterocycles with 4 to 8 atoms in the ring structure.

[0076] As used herein, the term "aryl" includes 3 to 12-membered substituted or unsubstituted monocyclic aryl groups, wherein each atom of the ring is a carbon (i.e., carbocyclic aryl) or one or more atoms are heteroatoms (i.e., heteroaryl). Aryl groups include 5 to 12-membered rings and 6 to 10-membered rings. The term "aryl" also includes polycyclic systems having two or more rings, wherein two or more carbons are common to two adjacent rings, and wherein at least one ring is aromatic; for example, the other rings may be cycloalkyl, cycloalkenyl, cycloynyl, aryl, heteroaryl, and / or heterocyclic. Carbocyclic aryl groups include benzene, naphthalene, phenanthrene, phenol, aniline, etc. Heteroaryl groups include substituted or unsubstituted aromatic 3 to 12-membered ring structures, 5 to 12-membered rings, and 5 to 10-membered rings, whose ring structures include 1 to 4 heteroatoms. Heteroaryl groups include, for example, pyrrole, furan, thiophene, imidazole, etc. Azoles, thiazoles, triazoles, pyrazoles, pyridines, pyrazines, pyridazines, and pyrimidines, etc. Aryl and heteroaryl groups can be monocyclic, bicyclic, or polycyclic. Each example of an aryl group may be independently and optionally substituted, i.e., unsubstituted (“unsubstituted aryl”) or substituted with one or more substituents (“substituted aryl”); for example, 1 to 5 substituents, 1 to 4 substituents, 1 to 3 substituents, 1 to 2 substituents, or only 1 substituent. The aromatic ring may be substituted at one or more ring positions by one or more substituents, such as halogens, azides, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, alkoxy, amino, nitro, mercapto, imino, amide, phosphonate / ester, hypophosphonate / ester, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyl, sulfonamide, ketone, aldehyde, ester, heterocyclic group, aromatic moiety or heteroaromatic moiety, fluoroalkyl (e.g., trifluoromethyl), cyano, etc. For example, in several embodiments, the aryl group may be an unsubstituted C5 to C6 group. 12 Aryl group, and in several embodiments, the aryl group may be a substituted C5 to C6 group. 10 Aryl.

[0077] As used herein, the terms “halogenated,” “halide,” or “halogen” mean halogen and include, but are not limited to, fluorine, chlorine, bromine, iodine, etc., in both radioactive and non-radioactive forms. In several embodiments, the halogen is selected from fluorine, chlorine, and bromine.

[0078] The term "heterocyclic group" or "heterocyclic group" refers to a 3- to 12-membered ring structure, a 5- to 12-membered ring, or a 5- to 10-membered ring, whose ring structure includes 1 to 4 heteroatoms. Heterocyclic rings can be monocyclic, bicyclic, spirocyclic, or polycyclic. Examples of heterocyclic groups include, for example, thiophene, thiathracene, furan, pyran, isobenzofuran, chromogenene, xanthine, and phenthiazolinone. (phenoxathiin), pyrrole, imidazole, pyrazole, isothiazole, isothiazole Azole, pyridine, pyrazine, pyrimidine, pyridazine, indoleazine, isoindole, indole, indazole, purine, quinazine, isoquinoline, quinoline, phthalazine, naphthidine, quinoxaline, quinazoline, cyclophosphine, pteridine, carbazole, caroline, phenanthridine, acridine, pyrimidine, phenanthroline, phenazine, phenpyrazine, phenothiazine, furan, phen Azides, pyrrolidines, oxopentane, thionylpentanes, Zazoles, piperidines, piperazines, morpholines, lactones, lactams such as azacyclobutanone and pyrrolidone, sulopentaamides, sulopentalides, etc. The heterocycle may be substituted at one or more positions with the aforementioned substituents, such as halogens, alkyl groups, aralkyl groups, alkenyl groups, alkynyl groups, cycloalkyl groups, hydroxyl groups, amino groups, nitro groups, mercapto groups, imino groups, amide groups, phosphate / ester groups, phosphonate / ester groups, hypophosphonate / ester groups, carbonyl groups, carboxyl groups, silyl groups, aminosulfonyl groups, sulfinyl groups, ethers, alkylthio groups, sulfonyl groups, ketones, aldehydes, esters, heterocyclic groups, aromatic or heteroaromatic moieties, -CF3, -CN, etc.

[0079] The term "carbonyl" is recognized in the art and includes a portion that can be represented by the following formula:

[0080]

[0081] Where X' is a bond or represents oxygen or sulfur, and R 15 Indicates hydrogen, alkyl, alkenyl, -(CH2) m -R 10 Or medicinal salt, R 16 Indicates hydrogen, alkyl, alkenyl, or -(CH2). m -R 10 , where m and R 10 As specified above. Where X' is oxygen and R... 15 or R 16 When it is not hydrogen, the formula represents "ester". When X' is oxygen and R... 15 As defined above, this part is referred to herein as a carboxyl group, and especially when R 15 When X' is hydrogen, the formula represents "carboxylic acid". When X' is oxygen and R... 16 When the oxygen atom is hydrogen, this formula represents "formate ester". Typically, when the oxygen atom in the above formula is replaced by sulfur, this formula represents "thiocarbonyl". When X' is sulfur and R... 15 or R 16 When it is not hydrogen, this formula represents a "thioester" group. In cases where X' is sulfur and R... 15 When X' is hydrogen, this formula represents a "thiocarboxylic acid" group. When X' is sulfur and R... 16 When it is hydrogen, the formula represents a "thiocarbamate" group. On the other hand, when X' is a bond and R... 15 When it is not hydrogen, the above formula represents a "ketone" group. When X' is a bond and R... 15 When it is hydrogen, the above formula represents an "aldehyde" group.

[0082] As used herein, the term “nitro” means -NO2; the term “halogen” means -F, -Cl, -Br or -I; the term “mercapto” means -SH; the term “hydroxyl” means -OH; the term “sulfonyl” means -SO2-; the term “azide” means -N3; ​​the term “cyano” means -CN; the term “isocyanate” means -NCO; the term “thiocyanate” means -SCN; the term “isothiocyanate” means -NCS; and the term “cyanate” means -OCN.

[0083] As is the case with each expression used herein (e.g., alkyl, m, n, etc.), when it appears more than once in any structure, it is intended to be independent of its definition elsewhere in the same structure.

[0084] The term "substituted" refers to a portion having a substituent that replaces hydrogen on one or more carbons of the main chain. It should be understood that "substituted" or "substituted with..." includes the implicit condition that such substitution meets the permissible valence of the substituted atom and substituent, and that the substitution produces a stable compound (e.g., one that does not spontaneously undergo transformations such as rearrangement, cyclization, elimination, etc.). As used herein, the term "substituted" is intended to include all permissible substituents in organic compounds. In a broad sense, permissible substituents include acyclic and cyclic, branched and linear, carbocyclic and heterocyclic, aromatic and non-aromatic substituents in organic compounds. For suitable organic compounds, permissible substituents can be one or more and can be the same or different. For the purposes of this disclosure, heteroatoms such as nitrogen may have hydrogen substituents and / or any permissible substituents of the organic compound described herein that satisfy the valence of that heteroatom. Substituents may include any substituents described herein, such as halogens, hydroxyl groups, carbonyl groups (e.g., carboxyl, alkoxycarbonyl, formyl, or acyl), thiocarbonyl groups (e.g., thioesters, thioacetates / esters, or thioformates / esters), alkoxy groups, phosphoryl groups, phosphates / esters, phosphonates / esters, phosphonites / esters, amino groups, amide groups, amidine groups, imine groups, cyano groups, nitro groups, azide groups, mercapto groups, alkylthio groups, sulfates / esters, sulfonates / esters, aminosulfonyl groups, sulfonamide groups, sulfonyl groups, heterocyclic groups, aralkyl groups, or aromatic or heteroaromatic moieties. In several embodiments, the substituents on the substituted alkyl group are selected from C10. 1-6 Alkyl, C 3-6Cycloalkyl, halogen, carbonyl, cyano, or hydroxyl groups. In many different embodiments, the substituent on the substituted alkyl group is selected from fluorine, carbonyl, cyano, or hydroxyl groups. Those skilled in the art will understand that the substituent itself may be substituted, if appropriate. Unless explicitly stated as “unsubstituted,” references to the chemical part herein should be understood to include substituted variants. For example, references to the “aryl” group or partly implicitly include both substituted and unsubstituted variants.

[0085] For the purposes of this disclosure, chemical elements were determined based on the periodic table, CAS version, Handbook of Chemistry and Physics, 67th edition, 1986-87, inner cover.

[0086] Exemplary compounds of this disclosure

[0087] In several embodiments, this disclosure relates to compounds of formula (I) or (II), or pharmaceutically acceptable salts thereof:

[0088] ,

[0089] in

[0090] X is NR 4 Or S;

[0091] R 1 It is H or -C(O)-C 1-10 alkyl;

[0092] R 2 It is a heterocyclic or aryl group;

[0093] R 3 It is a halogen, -CHF2 or -CF3;

[0094] R 4 It is H, -C 1-10 Alkyl, -C(O)-C 1-10 Alkyl, -C(O)-C 3-10 Cycloalkyl, -S(O)2-C 1-10 alkyl or

[0095] ;as well as

[0096] n is 0, 1, 2, 3 or 4.

[0097] In some embodiments, the compound is a compound of formula (I). In some embodiments, the compound is a compound of formula (II).

[0098] In some implementation schemes, R 1 It is H. In some implementations, R 1It is -C(O)-C 1-10 Alkyl group. In some embodiments, R 1 It is -C(O)-C1H3. In some implementations, R 1 It is -C(O)-C2H5. In some implementations, R 1 It is -C(O)-C3H7. In some implementations, R 1 It is -C(O)-C4H9.

[0099] In some implementation schemes, R 2 It is an unsubstituted heterocyclic group. In some embodiments, R 2 It is a substituted heterocyclic group. In some embodiments, the heterocyclic group is monocyclic. In some embodiments, the heterocyclic group is bicyclic. In some embodiments, the heterocyclic group is tricyclic. In some embodiments, the heterocyclic group is aromatic. In some embodiments, the heterocyclic group is non-aromatic. In some embodiments, R 2 It is an unsubstituted aryl group. In some embodiments, R 2 It is a substituted aryl group.

[0100] In some implementation schemes, R 2 Substitution with at least one of the following substituents: halogen, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, amino, nitro, mercapto, imino, amide, aminosulfonyl, sulfinyl, alkylthio, sulfonyl, ketone, heterocyclic, aromatic or heteroaromatic moiety, -CHF2-CF3, -CN. If R 2 If a substance is substituted by two or more substituents, the substituents may be the same or different.

[0101] In some implementation schemes, R 2 Selected from:

[0102] ,

[0103] .

[0104] In some implementation schemes, R 2 yes:

[0105] .

[0106] In some implementation schemes, R 2 yes:

[0107] .

[0108] In some implementation schemes, R 2 yes:

[0109] .

[0110] In some implementation schemes, R 2 yes:

[0111] .

[0112] In some implementation schemes, R 2 Selected from:

[0113]

[0114] .

[0115] In some implementation schemes, R 3 It is F. In some implementations, R 3 It is Cl. In some implementations, R 3 It is Br. In some implementations, R 3 It is -CHF2. In some implementations, R 3 It is -CF3.

[0116] In some implementation schemes, R 4 It is H. In some implementations, R 4 It is -C 1-10 Alkyl group. In some embodiments, R 4 It is methyl, ethyl, isopropyl, n-propyl, tert-butyl, isobutyl, or n-butyl. In some embodiments, R 4 It is a methyl group.

[0117] In some implementation schemes, R 4 It is -C(O)-C 1-10 Alkyl group. In some embodiments, R 4 It is -C(O)-methyl, -C(O)-ethyl, -C(O)-isopropyl, -C(O)-n-propyl, -C(O)-tert-butyl, -C(O)-isobutyl, or -C(O)-n-butyl. In some embodiments, R 4 It is -C(O)-C 5-10 alkyl.

[0118] In some implementation schemes, R 4 yes:

[0119] .

[0120] In some implementation schemes, R 4 It is -C(O)-C3H 7。

[0121] In some implementation schemes, R 4It is -C(O)-C 3-10 Cycloalkyl. In some embodiments, R 4 It is -C(O)-cyclopropyl. In some embodiments, R 4 It is -C(O)-cyclohexyl.

[0122] In some implementations, R 4 It is -S(O)2-C 1-10 Alkyl group. In some embodiments, R 4 It is -S(O)2-methyl, -S(O)2-ethyl, -S(O)2-isopropyl, -S(O)2-n-propyl, -S(O)2-tert-butyl, -S(O)2-isobutyl, or -S(O)2-n-butyl. In some embodiments, R 4 It is -S(O)2-C 5-10 alkyl.

[0123] In some implementation schemes, R 4 It is -S(O)2-C3H7.

[0124] In some implementation schemes, R 4 yes:

[0125] .

[0126] In some implementations, n is 0. In some implementations, n is 1. In some embodiments, n is 2. In some implementations, n is 3. In some implementations, n is 4.

[0127] In some embodiments, the compound of formula (I) is selected from:

[0128]

[0129] .

[0130] In some embodiments, the compound of formula (I) is:

[0131] .

[0132] In some embodiments, the compound of formula (I) is selected from:

[0133]

[0134] .

[0135] In some embodiments, the compound of formula (I) is:

[0136] .

[0137] In some embodiments, the compound of formula (II) is selected from:

[0138] .

[0139] In some embodiments, the compound of formula (I) is selected from...

[0140] .

[0141] In some embodiments, the compound of formula (I) is selected from:

[0142] .

[0143] In some embodiments, the compound of formula (I) is:

[0144] .

[0145] In some embodiments, the compound of formula (I) is

[0146] .

[0147] In some embodiments, the compound of formula (I) is

[0148] .

[0149] Exemplary pharmaceutical compositions

[0150] In several embodiments, this disclosure relates to pharmaceutical compositions comprising any of the compounds disclosed herein and a pharmaceutically acceptable carrier.

[0151] This includes, but is not limited to, treatment of human patients by administering an effective amount of the active compound or its pharmaceutically acceptable prodrug or salt (with a pharmaceutically acceptable carrier or diluent). The active substance may be administered via any suitable route, such as oral, parenteral, intravenous, intradermal, subcutaneous, or topical (in liquid or solid form).

[0152] The concentration of the active compound in a pharmaceutical composition will depend on the rates of absorption, inactivation, and excretion of the drug, as well as other factors known to those skilled in the art. It should be noted that dosage values ​​will also vary depending on the severity of the condition to be alleviated. It should also be understood that, for any particular subject, the specific dosage regimen should be adjusted over time based on individual needs and the professional judgment of the person administering or supervising the administration of the composition, and the concentration ranges listed herein are merely exemplary and not intended to limit the scope or practice of the claimed compositions. The active ingredient may be administered once or divided into many smaller doses administered at different time intervals.

[0153] In several embodiments, the active compound is administered orally. Oral compositions typically include an inert diluent or an edible carrier. They may be encapsulated in gelatin capsules or compressed into tablets. For oral therapeutic administration, excipients may be introduced into the active compound and administered in tablet, lozenge, or capsule form. Pharmaceutically compatible binders and / or excipients may be included as part of the composition.

[0154] Tablets, pills, capsules, lozenges, etc., may contain any of the following components or compounds with similar properties: binders (e.g., microcrystalline cellulose, tragacanth gum, or gelatin), excipients (e.g., starch or lactose), disintegrants (e.g., alginic acid, Primogel, or corn starch), lubricants (e.g., magnesium stearate or sterotes), gliding agents (e.g., colloidal silica), sweeteners (e.g., sucrose or saccharin), or flavoring agents (e.g., peppermint, methyl salicylate, or orange flavoring). When the dosage unit is in capsule form, in addition to the substances of the above types, it may also contain a liquid carrier, such as fatty oil. Furthermore, the dosage unit may contain a variety of other substances that alter the physical form of the dosage unit, such as sugar coating, shellac coating, or other enteric coatings.

[0155] The compound can be applied as an ingredient in elixirs, suspensions, syrups, wafers, chewing gum, etc. Syrups, in addition to the active compound, may also contain sucrose or sweeteners as sweeteners, as well as various preservatives, pigments and colorings, and flavoring agents.

[0156] The compound, or its pharmaceutically acceptable prodrug or salt, may also be mixed with other active substances that do not impair the desired effect or with substances that complement the desired effect, such as antibiotics, antifungals, anti-inflammatory agents, or other antiviral agents (including, but not limited to, nucleoside compounds). Solutions or suspensions for parenteral, intradermal, subcutaneous, or topical application may contain the following components: sterile diluents (e.g., water for injection, saline solution, non-volatile oils, polyethylene glycol, glycerin, propylene glycol, or other synthetic solvents), antibacterial agents (e.g., benzyl alcohol or methylparaben), antioxidants (e.g., ascorbic acid or sodium bisulfite), chelating agents (e.g., EDTA), buffers (e.g., acetates, citrates, or phosphates), and tonic agents (e.g., sodium chloride or glucose). Parenteral preparations may be packaged in ampoules, disposable syringes, or multi-dose vials (made of glass or plastic).

[0157] If administered intravenously, the carriers include normal saline and phosphate-buffered saline (PBS).

[0158] In several embodiments, the active compound is prepared together with a carrier that will protect the compound from rapid elimination from the body, such as a controlled-release formulation, including but not limited to implants and microencapsulated delivery systems. Biodegradable, biocompatible polymers such as ethylene vinyl acetate, polyanhydride, polyglycolic acid, collagen, polyorthoesters, and polylactic acid can be used. For example, enteric-coated compounds can be used to protect against degradation by gastric acid. Methods for preparing such formulations will be apparent to those skilled in the art. Suitable materials are also commercially available.

[0159] Liposome suspensions (including, but not limited to, liposomes targeting infected cells with monoclonal antibodies against viral antigens) are also pharmaceutically viable carriers. These can be prepared according to methods known to those skilled in the art, for example, as described in U.S. Patent No. 4,522,811 (incorporated by reference). For example, a liposome formulation can be prepared by dissolving a suitable lipid (e.g., stearoylphosphatidylethanolamine, stearoylphosphatidylcholine, arachidonicylphosphatidylcholine, and cholesterol) in an inorganic solvent, followed by evaporation to leave a dried lipid film on the surface of a container. An aqueous solution of the active compound is then introduced into the container. The container is then manually swirled to displace the lipid material from the sides of the container and disperse the lipid aggregates, thereby forming a liposome suspension.

[0160] Exemplary methods of this disclosure

[0161] In several embodiments, this disclosure relates to methods for treating pDC, B-cell, macrophage, or monocyte-mediated diseases or conditions, comprising the step of administering a therapeutically effective amount of any of the aforementioned compounds to a subject in need. In some embodiments, the pDC-mediated disease is lupus, Crohn's disease, irritable bowel syndrome (IBS), type 1 diabetes, dermatomyositis, Sjögren's syndrome, psoriasis, or any type 1 interferon-driven interferon disease. In some embodiments, the pDC-mediated disease is multiple sclerosis (MS). Example

[0162] The present disclosure, which is generally described herein, will be more readily understood by referring to the following examples, which are for illustrative purposes only and are not intended to limit the scope of the disclosure.

[0163] Example 1. General schemes and procedures for preparing the compounds disclosed herein. The synthesis of various compounds is described in... Figure 6A As shown in the image.

[0164] Example 2. Representative preparation of the compounds disclosed herein.

[0165] (A) Chemical materials

[0166] Chemicals and reagents were purchased from commercial suppliers (including Sigma-Aldrich, Fisher Scientific, Combi-Blocks, MedChemExpress, Alfa Aesar, and AstaTech) and were used as is unless otherwise specified, without further purification. Anhydrous solvents were purchased from Sigma-Aldrich's Sure / Seal™ formulation. All reactions were monitored by thin-layer chromatography (TLC, Merck silica gel 60 F-254 plates). Plates were stained with p-anisaldehyde (2.5% p-anisaldehyde, 1% AcOH, 3.5% H2SO4 (concentration) in 95% EtOH), ninhydrin (0.3% ninhydrin (w / v), 97:3 EtOH-AcOH), KMnO4 (1.5 g KMnO4, 10 g K2CO3, and 1.25 mL 10% NaOH in 200 mL water), or visualized directly under UV light. Reaction purification was performed using rapid chromatography (230 to 400 mesh silica gel), Biotage®, or preparative thin-layer chromatography (pTLC, Analtech, 500 to 2000 μm thickness). NMR spectra were recorded in the specified solvents on a Bruker DPX-400 or Bruker AV-600 spectrometer. Multiplicity was reported using the following abbreviations: s singlet; d doublet; t triplet; q quartet; p quintet; m multiplet; br broad peak; dd doublet; dt doublet; td triplet; chemical shifts were reported in ppm relative to the residual solvent peak, and J values ​​were reported in Hz. Mass spectrometry data were collected on an Agilent 6120 single quadrupole LC / MS instrument (ESI, low resolution).

[0167] (B) Compound synthesis and characterization data:

[0168] a) General Synthesis Scheme 1:

[0169]

[0170] General Operation 1: Coupling Operation for the Synthesis of Benzid[d]imidazolamine Intermediate (S1)

[0171] K₂CO₃ (3.0 equivalents) was added to a dry round-bottom flask containing a commercially available 2-aminobenzimidazole derivative (1.0 equivalent) and the corresponding aldehyde (1.0 equivalent) in anhydrous methanol, and the reaction mixture was heated at 50 °C for 16 to 30 hours. The solvent was filtered to remove excess potassium carbonate, and sodium triacetoxyborohydride (1.5 equivalent) was added to the solution at 0 °C, and the resulting mixture was stirred at room temperature for 3 to 5 hours. After completion (monitored by TLC), the solvent was removed by rotary evaporation, the crude mixture was diluted with water and washed with a saturated aqueous solution of NaHCO₃, extracted in ethyl acetate, the combined extracts were dried over Na₂SO₄, filtered and concentrated under vacuum, and purified by column chromatography on a biotage to give the corresponding amine (S1).

[0172] General procedure 2: Coupling of amine intermediate (S1) with acid

[0173] Commercially available butyric acid or 3-(3-(but-3-yn-1-yl)-3H-diazadiazino-3-yl)propionic acid (1.1 equivalents), DIPEA (3.0 equivalents), EDC-HCl (1.5 equivalents), and HOBt (1.5 equivalents) were added to vials containing the corresponding amine intermediate (S1, 1 equivalent) in DCM (60 mM relative to S1). The reaction mixture was stirred at room temperature for 4 hours to overnight (when TLC indicated completion). The crude mixture was diluted with DCM and washed first with saturated aqueous NH4Cl and saturated aqueous NaHCO3 solutions, then dried over anhydrous Na2SO4, and volatiles were removed by rotary evaporation. The crude product was purified by PTLC or rapid column chromatography to obtain the corresponding product.

[0174] General procedure 3: Coupling of amine intermediate (S1) with acid

[0175] A solution of the corresponding butyric acid or 3-(3-(but-3-yn-1-yl)-3H-diazapropidin-3-yl)propionic acid in DMF (60 mM relative to S1) was added at 0 °C with HATU (1.1 equivalents) and DIPEA (3.0 equivalents), and the resulting mixture was stirred for 5 min. The corresponding amine intermediate (S1) was then added, and the mixture was stirred at room temperature until TLC indicated complete depletion of the amine. The crude mixture was diluted with cold water and extracted with ethyl acetate, then dried over anhydrous Na2SO4 and volatiles were removed by rotary evaporation. The crude product was purified by PTLC or rapid column chromatography to obtain the corresponding product.

[0176] General Operation 4: Coupling Operation for the Synthesis of Amides Using Acyl Chlorides

[0177] Triethylamine (1.1 equivalents) was added to a solution of the corresponding amine (S1, 1.0 equivalents) in DCM (0.1 M), followed by the slow addition of the corresponding acyl chloride (1.0 equivalents) at 0 °C. The resulting mixture was stirred at room temperature until TLC indicated complete depletion of the amine. The crude mixture was diluted with DCM, washed first with saturated aqueous NH4Cl and saturated aqueous NaHCO3 solutions, then dried over anhydrous Na2SO4 and evaporated by rotary evaporation to remove volatiles. The crude product was purified by PTLC or Biotage® to obtain the corresponding product.

[0178] General Operation 5: Coupling Operations for Synthesizing N-alkyl Molecules

[0179] Anhydrous K₂CO₃ (2.0 equivalents) was added to a solution of the corresponding amine (S₁, 1.0 equivalent) in DMF (0.1 M), followed by the addition of the corresponding alkyl iodide or 3-(but-3-yn-1-yl)-3-(2-iodoethyl)-3H-diazidine (2.0 equivalents) at room temperature. The resulting mixture was stirred at 50 °C until the TLC-indicated amine was completely depleted, typically 18 to 24 hours. The crude mixture was diluted with cold water and extracted with ethyl acetate. The combined extracts were dried over anhydrous Na₂SO₄, and volatiles were removed by rotary evaporation. The crude product was purified by PTLC or rapid column chromatography to obtain the corresponding product.

[0180] b) General synthesis scheme 2:

[0181]

[0182] 6-Bromo-9-ethylcarbazole-3-carbaldehyde (S2) was prepared by dissolving 9-ethylcarbazole-3-carbaldehyde (2 g, 8.95 mmol) in DMF (15 mL) and cooling the solution in an ice bath. N-bromosuccinimide (1.91 g, 10.74 mmol) was added dropwise to the DMF (10 mL) solution over 10 minutes. The reaction mixture was stirred at room temperature for 2 hours. The mixture was then poured into ice water and extracted with ethyl acetate. The combined extracts were dried over anhydrous Na2SO4, and volatiles were removed by rotary evaporation. The crude product was purified by rapid column chromatography to give the corresponding 6-bromo-9-ethylcarbazole-3-carbaldehyde product (S2).

[0183] General Operation 6: Suzuki Coupling Operation for the Synthesis of (S3)

[0184] A solution of 6-bromo-9-ethylcarbazole-3-carbaldehyde (0.822 mmol), boric acid (0.986 mmol), and potassium carbonate (0.246 mmol) in dimethoxyethane (9 mL) and water (3 mL) was degassed by argon blowing for 5 min, followed by the addition of tetrakis(triphenylphosphine)palladium (47 mg, 0.041 mmol). The resulting mixture was stirred at 80 °C for 6 to 8 h. After cooling, the reaction mixture was filtered through diatomaceous earth, diluted with water, and extracted in ethyl acetate. The combined extracts were dried over anhydrous Na₂SO₄ and volatiles were removed by rotary evaporation. The crude product was purified by rapid column chromatography in ethyl acetate / hexane to obtain the corresponding product (S3).

[0185] c) General synthesis scheme 3:

[0186]

[0187] Step 1; Synthesis of intermediate (S4): A substituted cyclohexanone (1.1 equivalents) was added to a stirred solution of 4-bromophenylhydrazine (1.1 equivalents) in AcOH, and the reaction mixture was stirred at 118 °C for 3 hours. After cooling, acetic acid was removed by rotary evaporation. The reaction mixture was diluted with water and a saturated aqueous solution of NaHCO3 and extracted with ethyl acetate. The combined extracts were dried over anhydrous Na2SO4 and volatiles were removed by rotary evaporation. The crude product was purified by rapid column chromatography with ethyl acetate / hexane to obtain the corresponding product (S4).

[0188] Step 2; Synthesis of intermediate (S5): At 0°C, a suspension of sodium hydride (1.1 equivalents) (60% in mineral oil) was slowly added to a stirred solution of (S4) (1 equivalent) in DMF over 10 minutes. The resulting mixture was stirred in an ice bath for 15 minutes, followed by dropwise addition of iodoethane solution (1.5 equivalents) over 5 minutes. The resulting mixture was stirred at room temperature for 1 hour. The reaction mixture was diluted with water and extracted with ethyl acetate. The combined extracts were dried over anhydrous Na2SO4 and volatiles were removed by rotary evaporation. The crude product was purified by rapid column chromatography with ethyl acetate / hexane to obtain the corresponding product (S5).

[0189] Step 3; Synthesis of intermediate (S6): At -78°C under argon atmosphere, n-butyllithium (1.1 equivalents) was added to a stirred solution of (S5) (1 equivalent) in THF. The resulting mixture was stirred for 20 minutes, followed by the addition of DMF (3 equivalents). The reaction mixture was stirred at room temperature for 2 hours. The mixture was then poured into an ice-cold solution of ammonium chloride and extracted with ethyl acetate. The combined extracts were dried over anhydrous Na₂SO₄ and volatiles were removed by rotary evaporation. The crude product was purified by rapid column chromatography to obtain the corresponding aldehyde (S6).

[0190] d) General synthesis scheme 4:

[0191]

[0192] Step 1; Synthesis of intermediate (S7): At -78°C under argon atmosphere, n-butyllithium (1.1 equivalents) was added to a stirred solution of the corresponding wittig salt (1.0 equivalent) in THF. The resulting mixture was stirred for 20 minutes, followed by the addition of a solution of 9-ethyl-3-carbazole carboxaldehyde (1.0 equivalent) in THF. The reaction mixture was stirred at room temperature for 2 hours. The mixture was poured into an ice-cold solution of ammonium chloride and extracted with ethyl acetate. The combined extracts were dried over anhydrous Na₂SO₄ and volatiles were removed by rotary evaporation. The crude product was purified by rapid column chromatography to obtain the corresponding aldehyde (S7).

[0193] Step 2; Synthesis of intermediate (S8): 5% wt Pd / C was added to a solution of intermediate (S7) in methanol, and the resulting mixture was stirred at room temperature in a hydrogen atmosphere for 8 hours. After completion, the reaction mixture was filtered through a short diatomaceous earth pad and washed with methanol, dried by rotary evaporation, dissolved in DMF, and N-bromosuccinimide (1.2 equivalents) was added dropwise to the DMF solution over 10 minutes at 0°C. The reaction mixture was stirred at room temperature for 2 hours. The mixture was poured into ice water and extracted with ethyl acetate. The combined extracts were dried over anhydrous Na2SO4 and volatiles were removed by rotary evaporation. The crude product was purified by rapid column chromatography to obtain the corresponding brominated product (S8).

[0194] Step 3; Synthesis of intermediate (S9): At -78°C under argon atmosphere, n-butyllithium (1.1 equivalents) was added to a stirred solution of (S8) (1.0 equivalents) in THF. The resulting mixture was stirred for 20 minutes, followed by the addition of DMF (3.0 equivalents). The reaction mixture was stirred at room temperature for 2 hours. The mixture was then poured into an ice-cold solution of ammonium chloride and extracted with ethyl acetate. The combined extracts were dried over anhydrous Na₂SO₄ and volatiles were removed by rotary evaporation. The crude product was purified by rapid column chromatography to obtain the corresponding aldehyde (S9).

[0195]

[0196] 1-(2-(((1H-indol-5-yl)methyl)amino)-1H-benzo[d]imidazol-1-yl)but-1-one (AJ2-1): synthesized according to Scheme 1 and subsequent general procedure 4, purified by biotage (hexane / EtOAc, 6:4) to give AJ2-1 (17 mg, 62%) as a grayish-white solid.

[0197]

[0198] C 20 H 21 LCMS calculated value of N4O: 333.2 (M+H) + ), Measured value: 333.2.

[0199]

[0200] N-((1H-indol-5-yl)methyl)-N-(benzo[d]thiazo-2-yl)butyramide (AJ2-2): synthesized according to Scheme 1 and subsequent general operation 2, purified by PTLC (hexane / EtOAc, 4:2) to give AJ2-2 (8 mg, 62%) as a brown solid.

[0201]

[0202] C 20 H 20 N3OS LCMS calculated value: 350.1 (M+H) + ), Measured value: 350.0.

[0203]

[0204] N-((5-bromo-1H-indol-3-yl)methyl)-1H-benzo[d]imidazol-2-amine (AJ2-3A): synthesized according to Scheme 1, purified by biotage (hexane / EtOAc, 3:7) to give AJ2-3A (160 mg, 64%) as a light brown solid.

[0205]

[0206] C 16 H 14 LCMS calculated value of BrN4: 341.0 (M+H) + ), Measured value: 340.9.

[0207]

[0208] 1-(2-(((5-bromo-1H-indol-3-yl)methyl)amino)-1H-benzo[d]imidazol-1-yl)but-1-one (AJ2-3): synthesized according to Scheme 1 and general procedure 4, purified by biotage (hexane / EtOAc, 3:2) to give AJ2-3 (32 mg, 64%) as a brown solid.

[0209]

[0210] C20 H 20 LCMS calculated value of BrN4O: 411.1 (M+H) + ), Measured value: 411.1.

[0211]

[0212] 1-(2-((isoquinolin-5-ylmethyl)amino)-1H-benzo[d]imidazol-1-yl)but-1-one (AJ2-4): synthesized according to Scheme 1 and general procedure 4, purified by biotage (hexane / EtOAc, 3:2) to give AJ2-4 (7 mg, 54%) as a light brown solid.

[0213]

[0214] C 21 H 21 LCMS calculated value of N4O: 345.2 (M+H) + ), Measured value: 345.2.

[0215]

[0216] 1-(2-(((1H-pyrrolo[2,3-b]pyridin-3-yl)methyl)amino)-1H-benzo[d]imidazol-1-yl)but-1-one (AJ2-5): synthesized according to Scheme 1 and general procedure 4, purified by biotage (hexane / EtOAc, 4:2) to give AJ2-5 (12 mg, 62%) as a brown solid.

[0217]

[0218] C 19 H 20 LCMS calculated value of N5O: 334.1 (M+H) + ), Measured value: 334.1.

[0219]

[0220] 1-(2-((pyrazolo[1,5-a]pyridin-5-ylmethyl)amino)-1H-benzo[d]imidazol-1-yl)but-1-one (AJ2-6): synthesized according to Scheme 1 and general procedure 4, purified by biotage (hexane / EtOAc, 4:2) to give AJ2-6 (6 mg, 52%) as a brown solid.

[0221]

[0222]

[0223] C 19 H 20 LCMS calculated value of N5O: 334.16 (M+H) + ), Measured value: 334.16.

[0224]

[0225] 1-(2-((3,4-dimethoxybenzyl)amino)-1H-benzo[d]imidazol-1-yl)but-1-one (AJ2-7): synthesized according to Scheme 1 and general procedure 4, purified by biotage (hexane / EtOAc, 3:2) to give AJ2-7 (12 mg, 72%) as a brown solid.

[0226]

[0227] C 20 H 24 LCMS calculated value of N3O3: 354.2 (M+H) + ), Measured value: 354.2.

[0228]

[0229] 1-(2-(((1-benzyl-1H-indol-5-yl)methyl)amino)-1H-benzo[d]imidazol-1-yl)but-1-one (AJ2-8): synthesized according to Scheme 1 and general procedure 4, purified by biotage (hexane / EtOAc, 3:2) to give AJ2-8 (14 mg, 74%) as a grayish-white solid.

[0230]

[0231] C 27 H 27 LCMS calculated value of N4O: 423.2 (M+H) + ), Measured value: 423.2.

[0232]

[0233] 1-(2-(((1H-benzo[d]imidazol-5-yl)methyl)amino)-1H-benzo[d]imidazol-1-yl)but-1-one (AJ2-9): synthesized according to Scheme 1 and general procedure 4, and purified by PTLC (DCM / MeOH, 9:1) to give AJ2-9 (6 mg, 48%) as a grayish-white solid.

[0234]

[0235] C 19 H 20 LCMS calculated value of N5O: 334.2 (M+H) + ), Measured value: 334.16.

[0236]

[0237] (2-(((1H-indol-5-yl)methyl)amino)-1H-benzo[d]imidazol-1-yl)(cyclopropyl)methyl ketone (AJ2-10): synthesized according to Scheme 1 and general procedure 4, purified by PTLC (hexane / ethyl acetate 3:2) to give AJ2-10 (11 mg, 54%) as a grayish-white solid.

[0238]

[0239] C 20 H 19 LCMS calculated value of N4O: 331.1 (M+H) + ), Measured value: 331.1.

[0240]

[0241] N-((1H-indol-5-yl)methyl)-1-(propylsulfonyl)-1H-benzo[d]imidazol-2-amine (AJ2-11): synthesized according to Scheme 1 and general procedure 4, and purified by PTLC (hexane / ethyl acetate 3:2) to give AJ2-11 (5 mg, 43%) as a grayish-white solid.

[0242]

[0243]

[0244] C 19 H 21 LCMS calculated value of N4O2S: 369.1 (M+H) + ), Measured value: 469.13.

[0245]

[0246] 1-(2-(((1H-indol-5-yl)methyl)amino)-1H-benzi[d]imidazol-1-yl)-3,5,5-trimethylhex-1-one (AJ2-12): synthesized according to Scheme 1 and general procedure 4, purified by PTLC (hexane / ethyl acetate 3:2) to give AJ2-12 (6 mg, 45%) as a grayish-white solid.

[0247]

[0248] C 25 H 31 LCMS calculated value of N4O: 403.2 (M+H) + ), Measured value: 403.2.

[0249]

[0250] 1-(2-(((1H-indol-5-yl)methyl)amino)-1H-benzi[d]imidazol-1-yl)-3-cyclopentylprop-1-one (AJ2-13): synthesized according to Scheme 1 and general procedure 4, purified by PTLC (hexane / ethyl acetate 3:2) to give AJ2-13 (8 mg, 47%) as a grayish-white solid.

[0251]

[0252] C 24 H 27 LCMS calculated value of N4O: 387.2 (M+H) + ), Measured value: 387.2.

[0253]

[0254] (2-(((1H-indol-5-yl)methyl)amino)-1H-benzo[d]imidazol-1-yl)(cyclohexyl)methyl ketone (AJ2-14): synthesized according to Scheme 1 and subsequent general procedure 4, purified by PTLC (hexane / ethyl acetate 3:2) to give AJ2-14 (6 mg, 47%) as a grayish-white solid.

[0255]

[0256] C 23 H 25 LCMS calculated value of N4O: 373.2 (M+H) + ), Measured value: 373.2.

[0257]

[0258] 1-(2-(((1H-indol-5-yl)methyl)amino)-1H-benzi[d]imidazol-1-yl)-3-(3-(but-3-yn-1-yl)-3H-diazadiazin-3-yl)prop-1-one (AJ2-15): synthesized according to Scheme 1 and general procedure 1, purified by biotage (hexane / ethyl acetate 3:2) to give AJ2-15 (14 mg, 54%) as a colorless liquid.

[0259]

[0260] C 24 H 23 LCMS calculated value of N6O: 411.2 (M+H) + ), Measured value: 411.0.

[0261]

[0262] 1-(2-(((6-methoxypyridin-3-yl)methyl)amino)-1H-benzo[d]imidazol-1-yl)but-1-one (AJ2-16): synthesized according to Scheme 1 and general procedure 4, purified by biotage (hexane / ethyl acetate 3:2) to give AJ2-16 (12 mg, 47%) as a grayish-white solid.

[0263]

[0264] C 18 H 21 LCMS calculated value of N4O2: 325.1 (M+H) + ), Measured value: 325.0.

[0265]

[0266] 1-(2-(((5-fluoro-1H-indol-3-yl)methyl)amino)-1H-benzo[d]imidazol-1-yl)but-1-one (AJ2-17A): synthesized according to Scheme 1 and general procedure 1, purified by biotage (hexane / ethyl acetate 4:6) to give AJ2-17A (74 mg, 54%) as a brown solid.

[0267]

[0268] C 16 H 14 LCMS calculated value of FN4: 281.1 (M+H) + ), Measured value: 281.1.

[0269]

[0270] 1-(2-(((5-fluoro-1H-indol-3-yl)methyl)amino)-1H-benzo[d]imidazol-1-yl)but-1-one (AJ2-17): synthesized according to Scheme 1 and general procedure 4, purified by biotage (hexane / ethyl acetate 3:2) to give AJ2-17 (5 mg, 42%) as a grayish-white solid.

[0271]

[0272]

[0273] C 20 H 19 LCMS calculated value of FN4O: 351.2 (M+H) + ), Measured value: 351.0.

[0274]

[0275] N-((1H-indol-5-yl)methyl)-N-(2-oxo-1,2,3,4-tetrahydroquinolin-3-yl)butyramide (AJ2-18): synthesized according to Scheme 1 and general procedure 4, purified by biotage (hexane / ethyl acetate 6:4) to give AJ2-18 (8 mg, 47%) as a grayish-white solid;

[0276]

[0277] The LCMS calculated value of C22H24N3O2 is 362.1 (M+H+), and the measured value is also 362.1.

[0278]

[0279] 1-(2-((dibenzo[b,d]furan-4-ylmethyl)amino)-1H-benzo[d]imidazol-1-yl)but-1-one (AJ2-19): synthesized according to Scheme 1 and general procedure 4, purified by Biotage (hexane / ethyl acetate 3:2) to give AJ2-19 (14 mg, 62%) as a white solid.

[0280]

[0281] The LCMS calculated value of C24H22N3O2 is 384.2 (M+H+), and the measured value is also 384.2.

[0282]

[0283] 1-(2-(((5-(4-fluorophenyl)-1H-pyrazol-4-yl)methyl)amino)-1H-benzo[d]imidazol-1-yl)but-1-one (AJ2-20): synthesized according to Scheme 1 and general procedure 4, purified by PTLC (DCM / MeOH 9:1) to give AJ2-20 (6 mg, 42%) as a brown viscous liquid;

[0284]

[0285] The LCMS calculated value of C21H21FN5O is 378.1 (M+H+), and the measured value is 378.0.

[0286]

[0287] 1-(4-(((1-Butyryl-1H-benzo[d]imidazol-2-yl)amino)methyl)-5-(4-fluorophenyl)-1H-pyrazol-1-yl)but-1-one (AJ2-21): synthesized according to Scheme 1 and general procedure 4, purified by PTLC (DCM / MeOH 9.5:0.5) to give AJ2-21 (4 mg, 48%) as a brown viscous liquid;

[0288]

[0289] The LCMS calculated value of C25H27FN5O2 is 448.2 (M+H+), and the measured value is 448.1.

[0290]

[0291] N-((1H-indol-5-yl)methyl)-N-(2,3-dihydro-1H-indene-2-yl)butyramide (AJ2-22): synthesized according to Scheme 1 and general procedure 4, and purified by biotage (hexane / ethyl acetate 7:3) to give AJ2-22 (17 mg, 68%) as a brown viscous liquid.

[0292]

[0293] Note: Rotational isomers were observed. LCMS calculated value of C22H24N2O: 333.1 (M+H+), measured value: 333.1.

[0294]

[0295] N-((1H-benzo[d]imidazol-2-yl)methyl)-N-((1H-indol-5-yl)methyl)butyramide (AJ2-23): synthesized according to Scheme 1 and general procedure 4, and purified by PTLC to give AJ2-23 (13 mg, 57%) as a viscous liquid.

[0296]

[0297] The LCMS calculated value of C21H23N4O is 347.1 (M+H+), and the measured value is also 347.1.

[0298]

[0299] 1-(2-(((1H-indol-5-yl)methyl)amino)-5-fluoro-1H-benzo[d]imidazol-1-yl)but-1-one (AJ2-24): synthesized according to Scheme 1 and general procedure 4, purified by biotage (hexane / ethyl acetate 6:4) to give AJ2-24 (6 mg, 43%) as a viscous liquid;

[0300]

[0301]

[0302] The LCMS calculated value of C20H20FN4O is 351.1 (M+H+), and the measured value is also 351.1.

[0303]

[0304] N-((9-ethyl-9H-carbazole-3-yl)methyl)-1H-benzo[d]imidazol-2-amine (AJ2-25A): synthesized according to Scheme 1 and general procedure 1, purified by biotage (hexane / ethyl acetate 4:6) to give AJ2-25A (254 mg, 78%) as a yellow solid.

[0305]

[0306] The LCMS calculated value of C20H21FN4 is 341.1 (M+H+), and the measured value is 341.0.

[0307]

[0308] 1-(2-(((9-ethyl-9H-carbazole-3-yl)methyl)amino)-1H-benzo[d]imidazol-1-yl)but-1-one (AJ2-25): synthesized according to Scheme 1 and general procedure 4, purified by biotage (hexane / ethyl acetate 6:4) to give AJ2-25 (16 mg, 68%) as a yellow solid;

[0309]

[0310] The LCMS calculated value of C26H27N4O is 411.2 (M+H+), and the measured value is 411.0.

[0311]

[0312] N-((1H-indol-5-yl)methyl)-N-(2-(azacycloheptane-1-yl)-2-phenylethyl)butyramide (AJ2-26): synthesized according to Scheme 1 and general procedure 4, and purified by PTLC (hexane / ethyl acetate 7:3) to give AJ2-26 (22 mg, 74%) as a colorless liquid.

[0313]

[0314] Note: The LCMS calculated value of the rotational isomer C27H36N3O was 418.2 (M+H+), while the measured value was 418.1.

[0315]

[0316] 1-(2-(((1-phenyl-1H-pyrazol-4-yl)methyl)amino)-1H-benzo[d]imidazol-1-yl)but-1-one (AJ2-27): synthesized according to Scheme 1 and general procedure 4, purified by PTLC (hexane / ethyl acetate 6:4) to give AJ2-27 (16 mg, 62%) as a colorless liquid;

[0317]

[0318] The LCMS calculated value of C21H22N5O is 360.1 (M+H+), and the measured value is also 360.1.

[0319]

[0320] N-((5-bromo-1H-indol-3-yl)methyl)-1-methyl-1H-benzo[d]imidazol-2-amine A (AJ2-28): synthesized according to Scheme 1, purified by biotage (DCM / MeOH; 9:1) to give AJ2-28 (178 mg, 65%) as a brown solid.

[0321]

[0322] The calculated LCMS (ESI) value of C17H16BrN4 is 355.0 (M+H+), and the measured value is 354.9.

[0323]

[0324] N-((1H-indol-5-yl)methyl)-1-methyl-1H-benzo[d]imidazol-2-amine (AJ2-29): synthesized according to Scheme 1, purified by biotage (DCM / MeOH; 9:1) to give AJ2-29 (165 mg, 72%) as a brown solid.

[0325]

[0326] The LCMS calculated value of C17H17N4 is 277.1 (M+H+), and the measured value is also 277.1.

[0327]

[0328] N-((9-ethyl-9H-carbazole-3-yl)methyl)-1-methyl-1H-benzo[d]imidazol-2-amine (AJ2-30): synthesized according to Scheme 1, purified on biotage (hexane / ethyl acetate; 3:7) to give AJ2-30 (248 mg, 76%) as a yellow solid.

[0329]

[0330]

[0331] The LCMS calculated value of C23H23N4 is 355.1 (M+H+), and the measured value is also 355.1.

[0332]

[0333] N-((9-ethyl-9H-carbazole-3-yl)methyl)-N-(1-methyl-1H-benzo[d]imidazol-2-yl)butyramide (AJ2-31): synthesized according to Scheme 1 and general procedure 3, purified on biotage (DCM / MeOH; 9.5:0.5) to give AJ2-31 (64 mg, 52%) as a white solid.

[0334]

[0335] Note: The LCMS calculated value of the rotational isomer C27H28N4O is 425.2 (M+H+), and the measured value is 425.1.

[0336]

[0337] (3-(3-(but-3-yn-1-yl)-3H-diazapropidin-3-yl)-N-(9-ethyl-9H-carbazole-3-yl)methyl)-N-(1-methyl-1H-benzo[d]imidazol-2-yl)propionamide (AJ2-32): synthesized according to Scheme 1 and general procedure 3, purified by biotage (DCM / MeOH; 9.5:0.5) to give AJ2-32 (12 mg, 46%) as a light brown viscous liquid.

[0338]

[0339]

[0340] Note: The LCMS calculated value of the rotational isomer C31H31N6O is 503.2 (M+H+), and the measured value is 503.0.

[0341]

[0342] N-((5-bromo-1H-indol-3-yl)methyl)-3-(3-(but-3-yn-1-yl)-3H-diazapropidin-3-yl)-N-(1-methyl-1H-benzo[d]imidazol-2-yl)propionamide (AJ2-32): synthesized according to Scheme 1 and general procedure 3, purified by PTLC (DCM / MeOH; 9.5:0.5) to give AJ2-33 (4 mg, 27%) as a light brown viscous liquid.

[0343]

[0344] Note: The LCMS calculated value of the rotational isomer C25H24BrN6O was 503.1 (M+H+), and the measured value was 503.0.

[0345]

[0346] 1-(2-(3-(but-3-yn-1-yl)-3H-diazadiazino-3-yl)ethyl)-N-((9-ethyl-9H-carbazole-3-yl)methyl)-1H-benzo[d]imidazol-2-amine (AJ2-34): synthesized according to Scheme 1 and general procedure 5, purified by biotage (hexane / ethyl acetate; 4:6) to give AJ2-34 (43 mg, 64%) as a yellow viscous liquid;

[0347]

[0348]

[0349] The LCMS calculated value of C29H29N6 is 461.2 (M+H+), and the measured value is 461.0.

[0350]

[0351] 3-(3-(but-3-yn-1-yl)-3H-diazadiazino-3-yl)-1-(2-(((6-methoxypyridin-3-yl)methyl)amino)-1H-benzo[d]imidazol-1-yl)prop-1-one (AJ2-35): synthesized according to Scheme 1 and general procedure 3, purified by biotage (hexane / ethyl acetate; 4:6) to give AJ2-35 (12 mg, 62%) as a light brown viscous liquid.

[0352]

[0353] The LCMS calculated value of C22H23N6O2 is 403.1 (M+H+), and the measured value is 403.0.

[0354]

[0355] N-((1H-indol-5-yl)methyl)-3-(3-(but-3-yn-1-yl)-3H-diazapropidin-3-yl)-N-(2,3-dihydro-1H-indene-2-yl)acrylamide (AJ2-36): synthesized according to Scheme 1 and general procedure 2, purified by biotage (hexane / ethyl acetate; 4:6) to give AJ2-36 (12 mg, 46%) as a light brown viscous liquid.

[0356]

[0357] Note: Rotational isomers were observed. LCMS calculated value of C26H27N4O: 411.2 (M+H+), measured value: 411.2.

[0358]

[0359] 1-(2-(((5-bromo-1H-indol-3-yl)methyl)amino)-1H-benzo[d]imidazol-1-yl)-4,4,4-trifluorobut-1-one (AJ2-37): synthesized according to Scheme 1 and general procedure 4, purified on biotage (DCM / MeOH; 9.5:0.5) to give AJ2-37 (12 mg, 46%) as a light brown viscous liquid;

[0360]

[0361] The LCMS calculated value of C20H17BrF3N4O is 465.0 (M+H+), and the measured value is 466.8.

[0362]

[0363] N-((1,6,7,8-tetrahydrocyclopentano[g]indol-3-yl)methyl)-1H-benzo[d]imidazol-2-amine (AJ2-38): synthesized according to Scheme 1, purified by biotage (DCM / MeOH; 9.5:0.5) to give AJ2-38 (64 mg, 72%) as a grayish-white solid.

[0364]

[0365] The LCMS calculated value of C19H19N4 is 303.1 (M+H+), and the measured value is also 303.1.

[0366]

[0367] 1-(2-((((1,6,7,8-tetrahydrocyclopentano[g]indol-3-yl)methyl)amino)-1H-benzo[d]imidazol-1-yl)but-1-one (AJ2-39): synthesized according to Scheme 1 and general procedure 4, purified by biotage (hexane / ethyl acetate; 5:5) to give AJ2-39 (64 mg, 72%) as a white solid;

[0368]

[0369] The LCMS calculated value of C23H25N4O is 373.2 (M+H+), and the measured value is 373.1.

[0370]

[0371] (S)-N-(1H-benzo[d]imidazol-2-yl)-2-(6-methoxynaphthyl-2-yl)acrylamide (AJ2-40): synthesized according to Scheme 1 and general procedure 2, purified on biotage (hexane / ethyl acetate; 3:7) to give AJ2-40 (67 mg, 68%) as a white solid.

[0372]

[0373] The LCMS calculated value of C21H20N3O2 is 346.1 (M+H+), and the measured value is 346.0.

[0374]

[0375] (S)-N-(2-(6-methoxynaphthyl-2-yl)propyl)-1H-benzo[d]imidazol-2-amine (AJ2-41): synthesized according to Scheme 1, purified on biotage (hexane / ethyl acetate; 3:7) to give AJ2-41 (22 mg, 57%) as a light brown solid.

[0376]

[0377] The LCMS calculated value of C21H22N3O is 332.1 (M+H+), and the measured value is also 332.1.

[0378]

[0379] (S)-1-(2-((2-(6-methoxynaphthyl-2-yl)propyl)amino)-1H-benzo[d]imidazol-1-yl)but-1-one (AJ2-42): synthesized according to Scheme 1 and general procedure 4, purified on biotage (hexane / ethyl acetate; 6:4) to give AJ2-42 (22 mg, 57%) as a light brown solid.

[0380]

[0381] The LCMS calculated value of C25H28N3O2 is 402.2 (M+H+), and the measured value is 402.1.

[0382]

[0383] N-(1H-benzo[d]imidazol-2-yl)-2-(1-(4-chlorobenzoyl)-5-methoxy-2-methyl-1H-indol-3-yl)acetamide (AJ2-43): synthesized according to Scheme 1, purified on biotage (hexane / ethyl acetate; 4:6) to give AJ2-43 (42 mg, 64%) as a light brown solid.

[0384]

[0385] The LCMS calculated value of C26H22ClN4O3 is 473.1 (M+H+), and the measured value is 472.9.

[0386]

[0387] N-(1H-benzo[d]imidazol-2-yl)-2-(6-chloro-9H-carbazol-3-yl)acrylamide (AJ2-44): synthesized according to general procedure 2, purified on biotage (hexane / ethyl acetate; 4:6) to give AJ2-44 (24 mg, 54%) as a grayish-white solid.

[0388]

[0389] The LCMS calculated value of C22H18ClN4O is 389.1 (M+H+), and the measured value is 389.0.

[0390]

[0391] N-(2-(6-chloro-9H-carbazol-3-yl)propyl)-1H-benzo[d]imidazol-2-amine (AJ2-45): synthesized according to Scheme 1 and general procedure 1, purified on biotage (hexane / ethyl acetate; 3:7) to give AJ2-45 (8 mg, 42%) as a brown solid.

[0392]

[0393] The LCMS calculated value of C22H20ClN4 is 375.1 (M+H+), and the measured value is also 375.1.

[0394]

[0395] 3-(((1H-benzo[d]imidazol-2-yl)amino)methyl)-1H-indol-6-carboxynitrile (AJ2-46): synthesized according to Scheme 1 and general procedure 1, purified on biotage (hexane / ethyl acetate; 3:7) to give AJ2-46 (35 mg, 58%) as a brown solid.

[0396]

[0397] The LCMS calculated value of C17H14N5 is 288.1 (M+H+), and the measured value is also 288.1.

[0398]

[0399] N-((5-chloro-1H-indol-3-yl)methyl)-1H-benzo[d]imidazol-2-amine (AJ2-47): synthesized according to Scheme 1 and general procedure 1, purified on biotage (hexane / ethyl acetate; 3:7) to give AJ2-47 (43 mg, 64%) as a brown solid.

[0400]

[0401] The LCMS calculated value of C16H14ClN4 is 297.0 (M+H+), and the measured value is also 297.0.

[0402]

[0403] 9-Ethyl-6-(((1-Methyl-1H-benzo[d]imidazol-2-yl)amino)methyl)-9H-carbazole-3-carboxynitrile (AJ2-48): synthesized according to Scheme 1 and general procedure 1, purified on biotage (hexane / ethyl acetate; 3:7) to give AJ2-48 (32 mg, 65%) as a brown solid;

[0404]

[0405] The LCMS calculated value of C24H22N5 is 380.1 (M+H+), and the measured value is also 380.1.

[0406]

[0407] N-((9-ethyl-3,3-dimethyl-2,3,4,9-tetrahydro-1H-carbazol-6-yl)methyl)-1-methyl-1H-benzo[d]imidazol-2-amine (AJ2-49): synthesized according to Scheme 3 and general procedure 1, purified on biotage (hexane / ethyl acetate; 3:7) to give AJ2-49 (32 mg, 65%) as a brown solid;

[0408]

[0409]

[0410] The LCMS calculated value of C25H31N4 is 387.2 (M+H+), and the measured value is 387.1.

[0411]

[0412] N-((9-ethyl-3,3-dimethyl-2,3,4,9-tetrahydro-1H-carbazol-6-yl)methyl)-1H-benzo[d]imidazol-2-amine (AJ2-50): synthesized according to Scheme 3 and general procedure 1, purified on biotage (hexane / ethyl acetate; 6:4) to give AJ2-50 (45 mg, 62%) as a brown solid;

[0413]

[0414] The LCMS calculated value of C24H29N4 is 373.2 (M+H+), and the measured value is 373.1.

[0415]

[0416] 1-Butyl-N-((9-ethyl-3,3-dimethyl-2,3,4,9-tetrahydro-1H-carbazol-6-yl)methyl)-1H-benzo[d]imidazol-2-amine (AJ2-51): synthesized according to Scheme 3 and general procedure 1 and thereafter general procedure 4, purified on biotage (hexane / ethyl acetate; 6:4) to give AJ2-51 (22 mg, 56%) as a brown solid.

[0417]

[0418]

[0419] The LCMS calculated value of C28H37N4 is 428.2 (M+H+), and the measured value is 429.2.

[0420]

[0421] 1-Butyl-N-((9-ethyl-9H-carbazole-3-yl)methyl)-1H-benzo[d]imidazol-2-amine (AJ2-52): synthesized according to general procedure 1 and thereafter general procedure 5, purified on biotage (hexane / ethyl acetate; 5:5) to give AJ2-52 (18 mg, 62%) as a yellow solid.

[0422]

[0423] The LCMS calculated value of C26H29N4 is 397.2 (M+H+), and the measured value is also 397.2.

[0424]

[0425] N-((1H-indol-5-yl)methyl)-1-butyl-1H-benzo[d]imidazol-2-amine (AJ2-53): synthesized according to general procedure 1 and thereafter general procedure 5, purified on biotage (hexane / ethyl acetate; 6:4) to give AJ2-53 (16 mg, 56%) as a light brown liquid.

[0426]

[0427]

[0428] The LCMS calculated value of C20H23N4 is 319.1 (M+H+), and the measured value is also 319.1.

[0429]

[0430] 1-Butyl-N-(dibenzo[b,d]furan-4-ylmethyl)-1H-benzo[d]imidazol-2-amine (AJ2-54): synthesized according to general procedure 1 and thereafter general procedure 4, purified on biotage (hexane / ethyl acetate; 6:4) to give AJ2-54 (24 mg, 68%) as a light brown liquid;

[0431]

[0432] The LCMS calculated value of C24H24N3O is 370.1 (M+H+), and the measured value is also 370.1.

[0433]

[0434] 1-Butyl-N-((6-methoxypyridin-3-yl)methyl)-1H-benzo[d]imidazol-2-amine (AJ2-55): synthesized according to general procedure 1 and thereafter general procedure 4, purified on biotage (hexane / ethyl acetate; 4:6) to give AJ2-55 (18 mg, 62%) as a light brown liquid.

[0435]

[0436] The LCMS calculated value of C18H23N4O is 311.2 (M+H+), and the measured value is also 311.2.

[0437]

[0438] N-(benzo[d][1,3]m-dioxacyclopenten-5-ylmethyl)-1-butyl-1H-benzo[d]imidazol-2-amine (AJ2-56): synthesized according to general procedure 1 and thereafter general procedure 4, purified on biotage (hexane / ethyl acetate; 4:6) to give AJ2-56 (8 mg, 56%) as a light brown liquid;

[0439]

[0440] The LCMS calculated value of C19H22N3O2 is 324.1 (M+H+), and the measured value is also 324.1.

[0441]

[0442] 1-(2-(diethylamino)ethyl)-N-((9-ethyl-9H-carbazol-3-yl)methyl)-1H-benzo[d]imidazol-2-amine (AJ2-57): synthesized according to general procedure 1, purified on biotage (DCM / methanol; 9.5:0.5) to give AJ2-57 (35 mg, 74%) as a yellow solid.

[0443]

[0444] The LCMS calculated value of C28H34N5 is 440.2 (M+H+), and the measured value is 440.1.

[0445]

[0446] N-((5-bromo-1H-indol-3-yl)methyl)-1-butyl-1H-benzo[d]imidazol-2-amine (AJ2-58): synthesized according to Scheme 1 and general procedure 5, purified on biotage (hexane / ethyl acetate; 4:6) to give AJ2-58 (6 mg, 43%) as a light brown liquid;

[0447]

[0448] The LCMS calculated value of C20H22BrN4 is 397.0 (M+H+), and the measured value is also 397.0.

[0449]

[0450] N-((9-ethyl-9H-carbazole-3-yl)methyl)-1-phenethyl-1H-benzo[d]imidazol-2-amine (AJ2-59): synthesized according to Scheme 1 and general procedure 5, purified on biotage (hexane / ethyl acetate; 6:4) to give AJ2-59 (12 mg, 47%) as a pale yellow liquid.

[0451]

[0452] The LCMS calculated value of C30H29N4 is 445.2 (M+H+), and the measured value is 445.1.

[0453]

[0454] 1-Benzyl-N-((9-ethyl-9H-carbazole-3-yl)methyl)-1H-benzo[d]imidazol-2-amine (AJ2-60): synthesized according to general procedure 1, purified on biotage (hexane / ethyl acetate; 6:4) to give AJ2-60 (43 mg, 67%) as a pale yellow solid.

[0455]

[0456] The LCMS calculated value of C29H27N4 is 431.2 (M+H+), and the measured value is 431.1.

[0457]

[0458] N-((9-ethyl-3-(trifluoromethyl)-2,3,4,9-tetrahydro-1H-carbazol-6-yl)methyl)-1-methyl-1H-benzo[d]imidazol-2-amine (AJ2-61): synthesized according to Scheme 3 and general procedure 1, purified on biotage (hexane / ethyl acetate; 5:5) to give AJ2-61 (24 mg, 72%) as a brown solid.

[0459]

[0460]

[0461] The LCMS calculated value of C24H26F3N4 is 427.2 (M+H+), and the measured value is also 427.2.

[0462]

[0463] N-((9-ethyl-3-(trifluoromethyl)-2,3,4,9-tetrahydro-1H-carbazol-6-yl)methyl)-1H-benzo[d]imidazol-2-amine (AJ2-62): synthesized according to Scheme 3 and general procedure 1, purified on biotage (hexane / ethyl acetate; 5:5) to give AJ2-62 (34 mg, 63%) as a brown solid.

[0464]

[0465] The LCMS calculated value of C23H24F3N4 is 413.1 (M+H+), and the measured value is also 413.1.

[0466]

[0467] N-(1H-benzo[d]imidazol-2-yl)-9-ethyl-N-methyl-9H-carbazole-3-carboxamide (AJ2-63): synthesized according to general procedure 3, purified on biotage (hexane / ethyl acetate; 6:4) to give AJ2-63 (34 mg, 63%) as a brown solid.

[0468]

[0469] The LCMS calculated value of C23H21N4O is 369.1 (M+H+), and the measured value is also 369.1.

[0470]

[0471] 1-Butyl-N-((9-ethyl-3-(trifluoromethyl)-2,3,4,9-tetrahydro-1H-carbazole-6-yl)methyl)-1H-benzo[d]imidazol-2-amine (AJ2-64): synthesized according to Scheme 3 and subsequent general procedure 5, purified on biotage (hexane / ethyl acetate; 6:4) to give AJ2-64 (42 mg, 64%) as a brown solid.

[0472]

[0473] The LCMS calculated value of C27H32F3N4 is 369.2 (M+H+), and the measured value is also 369.2.

[0474]

[0475] N-((5-bromo-1H-indol-3-yl)methyl)-1-(2-(diethylamino)ethyl)-1H-benzo[d]imidazol-2-amine (AJ2-65): synthesized according to general scheme 3, purified on biotage (hexane / ethyl acetate; 4:6) to give AJ2-65 (24 mg, 66%) as a yellow solid.

[0476]

[0477] The LCMS calculated value of C22H27BrN5 is 440.1 (M+H+), and the measured value is also 440.1.

[0478]

[0479] 1-Benzyl-N-((5-bromo-1H-indol-3-yl)methyl)-1H-benzo[d]imidazol-2-amine (AJ2-66): synthesized according to general scheme 3, purified on biotage (hexane / ethyl acetate; 6:4) to give AJ2-66 (48 mg, 63%) as a brown solid.

[0480]

[0481] The LCMS calculated value of C23H20BrN4 is 431.0 (M+H+), and the measured value is also 431.0.

[0482]

[0483] (R)-1-(1H-benzo[d]imidazol-2-yl)-N-((9-ethyl-9H-carbazol-3-yl)methyl)ethane-1-amine (AJ2-67): synthesized according to general procedure 1, purified on biotage (hexane / ethyl acetate; 6:4) to give AJ2-67 (34 mg, 63%) as a brown solid.

[0484]

[0485] The LCMS calculated value of C24H25N4 is 369.2 (M+H+), and the measured value is also 369.2.

[0486]

[0487] N-((9-ethyl-9H-carbazole-3-yl)methyl)-1-(4,4,4-trifluorobutyl)-1H-benzo[d]imidazol-2-amine (AJ2-68): synthesized according to Scheme 1 and general procedure 5, purified on biotage (hexane / ethyl acetate; 6:4) to give AJ2-68 (34 mg, 63%) as a brown solid.

[0488]

[0489] The LCMS calculated value of C26H26F3N4 is 451.2 (M+H+), and the measured value is also 451.2.

[0490]

[0491] 3-(((1H-benzo[d]imidazol-2-yl)amino)methyl)-1H-indole-5-carboxylic acid methyl ester (AJ2-69): synthesized according to general procedure 1, purified on biotage (hexane / ethyl acetate; 3:7) to give AJ2-69 (30 mg, 23%) as a brown solid.

[0492]

[0493] The LCMS calculated value of C18H17N4O2 is 321.1 (M+H+), and the measured value is also 321.1.

[0494]

[0495] 3-(((1H-benzo[d]imidazol-2-yl)amino)methyl)-1H-indole-5-carboxylic acid (AJ2-70): Lithium hydroxide (0.023 mmol) was added to a solution of methyl 3-(((1H-benzo[d]imidazol-2-yl)amino)methyl)-1H-indole-5-carboxylic acid (AJ2-69) (0.078 mmol) in tetrahydrofuran (1 mL) and water (1 mL) at 0 °C. The reaction mixture was stirred at room temperature for 6 hours. After the reaction mixture was stirred, it was neutralized with 1 N·HCl (pH 6 to 7), and the solid was filtered to give AJ2-70 (6 mg, 42%) as a brown solid.

[0496]

[0497] The LCMS calculated value of C17H15N4O2 is 307.1 (M+H+), and the measured value is also 307.1.

[0498]

[0499] N-((1H-benzo[g]indol-3-yl)methyl)-1H-benzo[d]imidazol-2-amine (AJ2-71): synthesized according to general procedure 1, purified on biotage (hexane / ethyl acetate; 2:8) to give AJ2-71 (34 mg, 63%) as a grayish-white solid.

[0500]

[0501] The LCMS calculated value of C20H17N4 is 313.1 (M+H+), and the measured value is also 313.1.

[0502]

[0503] N-((1H-benzo[g]indol-3-yl)methyl)-1-methyl-1H-benzo[d]imidazol-2-amine (AJ2-72): synthesized according to general procedure 1, purified on biotage (hexane / ethyl acetate; 3:7) to give AJ2-62 (34 mg, 63%) as a brown solid.

[0504]

[0505]

[0506] The LCMS calculated value of C21H19N4 is 327.1 (M+H+), and the measured value is also 327.1.

[0507]

[0508] N-((9-ethyl-9H-carbazole-3-yl)methyl)quinoline-3-amine (AJ2-73): synthesized according to general procedure 1, purified on biotage (hexane / ethyl acetate; 6:4) to give AJ2-73 (76 mg, 71%) as a grayish-white solid.

[0509]

[0510] The LCMS calculated value of C24H22N3 is 352.2 (M+H+), and the measured value is also 352.2.

[0511]

[0512] N-((9-ethyl-9H-carbazole-3-yl)methyl)-N-(quinoline-3-yl)butyramide (AJ2-74): synthesized according to general procedure 1 and subsequently general procedure 4, purified by PTLC (hexane / ethyl acetate; 6:4) to give AJ2-62 (17 mg, 68%) as a brown solid.

[0513]

[0514] The LCMS calculated value of C28H28N3O is 422.2 (M+H+), and the measured value is 422.1.

[0515]

[0516] 1-(2-(3-(but-3-yn-1-yl)-3H-diazapropidin-3-yl)ethyl)-N-(3,4-dimethoxybenzyl)-1Hbenzi[d]imidazol-2-amine (AJ2-75): synthesized according to general procedure 1 and subsequently general procedure 5, purified by PTLC (hexane / ethyl acetate; 6:4) to give AJ2-75 (12 mg, 64%) as a brown solid.

[0517]

[0518] The LCMS calculated value of C23H26N5O2 is 404.2 (M+H+), and the measured value is also 404.2.

[0519]

[0520] N-((7-ethyl-7H-benzo[c]carbazole-10-yl)methyl)-1H-benzo[d]imidazol-2-amine (AJ2-76): synthesized according to general procedure 1, purified by biotage (hexane / ethyl acetate; 2:8) to give AJ2-76 (27 mg, 56%) as a brown solid.

[0521]

[0522] The LCMS calculated value of C26H23N4 is 391.1 (M+H+), and the measured value is also 391.1.

[0523]

[0524] N-((7-ethyl-7H-benzo[c]carbazole-10-yl)methyl)-1-methyl-1H-benzo[d]imidazol-2-amine (AJ2-77): synthesized according to general procedure 1, purified by biotage (hexane / ethyl acetate; 3:7) to give AJ2-77 (48 mg, 66%) as a brown solid.

[0525]

[0526] The LCMS calculated value of C27H25N4 is 405.2 (M+H+), and the measured value is 405.1.

[0527]

[0528] N-((9-(4-(tert-butyl)phenyl)-9H-carbazole-3-yl)methyl)-1-methyl-1H-benzo[d]imidazol-2-amine (AJ2-78): synthesized according to general procedure 1, purified by biotage (hexane / ethyl acetate; 3:7) to give AJ2-78 (65 mg, 72%) as a brown solid.

[0529]

[0530] The calculated LCMS value of C31H31N4 is 459.2 (M+H+), and the measured value is 459.1.

[0531]

[0532] N-((6-(3,5-dimethylisocyanate) (Azol-4-yl)-9-ethyl-9H-carbazole-3-yl)methyl)-1H-benzo[d]imidazol-2-amine (AJ2-79): synthesized according to general synthetic scheme 2 and subsequently by general operation 1, purified by biotage (hexane / ethyl acetate; 3:7) to give AJ2-79 (10 mg, 52%) as a brown solid.

[0533]

[0534] The LCMS calculated value of C27H26N5O is 436.2 (M+H+), and the measured value is 436.1.

[0535]

[0536] N-((6-(3,5-dimethylisocyanate) (Azol-4-yl)-9-ethyl-9H-carbazole-3-yl)methyl)-1-methyl-1H-benzo[d]imidazol-2-amine (AJ2-80): synthesized according to general procedure 2 and subsequently general procedure 1, purified by biotage (hexane / ethyl acetate; 3:7) to give AJ2-80 (34 mg, 62%) as a brown solid.

[0537]

[0538] The LCMS calculated value of C28H28N5O is 450.2 (M+H+), and the measured value is also 450.2.

[0539]

[0540] N-((6-bromo-9-ethyl-9H-carbazole-3-yl)methyl)-1-methyl-1H-benzo[d]imidazol-2-amine (AJ2-81): synthesized according to general procedure 1, purified by biotage (hexane / ethyl acetate; 3:7) to give AJ2-81 (85 mg, 76%) as a brown solid.

[0541]

[0542] The LCMS calculated value of C23H22BrN4 is 433.0 (M+H+), and the measured value is also 433.0.

[0543]

[0544] N-((6-(benzo[d][1,3]m-dioxacyclopenten-5-yl)-9-ethyl-9H-carbazole-3-yl)methyl)-1-methyl-1H-benzo[d]imidazol-2-amine (AJ2-82): synthesized according to general scheme 2 and general operation 1, purified by biotage (hexane / ethyl acetate; 2:8) to give AJ2-82 (13 mg, 52%) as a brown solid.

[0545]

[0546] The LCMS calculated value of C30H27N4O2 is 475.2 (M+H+), and the measured value is 475.1.

[0547]

[0548] 3-(9-ethyl-6-(((1-methyl-1H-benzo[d]imidazol-2-yl)amino)methyl)-9H-carbazole-3-yl)phenol (AJ2-83): synthesized according to general scheme 2 and general procedure 1, purified by biotage (hexane / ethyl acetate; 3:7) to give AJ2-83 (35 mg, 62%) as a grayish-white solid.

[0549]

[0550] The LCMS calculated value of C29H27N4O is 447.2 (M+H+), and the measured value is 447.1.

[0551]

[0552] 2-((9-ethyl-2,3,4,9-tetrahydro-1H-carbazole-6-yl)methyl)isoindoline-1,3-dione (AJ2-83): synthesized according to general procedure 4, purified by biotage (hexane / ethyl acetate; 6:4) to give AJ2-85 (45 mg, 62%) as a yellow solid.

[0553]

[0554] The LCMS calculated value of C23H23N2O2 is 359.1 (M+H+), and the measured value is also 359.1.

[0555]

[0556] 3-(3-(but-3-yn-1-yl)-3H-diazadiazidin-3-yl)-N-cyclopentyl-N-((9-ethyl-9H-carbazole-2-yl)methyl)acrylamide (AJ2-86): synthesized according to general procedure 2, purified by biotage (hexane / ethyl acetate; 6:4) to give AJ2-86 (14 mg, 62%) as a colorless oil. Note: rotational isomers were observed.

[0557]

[0558] Note: The LCMS calculated value of the rotational isomer C28H33N4O was 441.2 (M+H+), while the measured value was 441.1.

[0559]

[0560] N-((9-ethyl-6-methoxy-9H-carbazole-3-yl)methyl)-1-methyl-1H-benzo[d]imidazol-2-amine (AJ2-87): synthesized according to general procedure 1, purified by biotage (hexane / ethyl acetate; 4:6) to give AJ2-87 (72 mg, 64%) as a yellow solid.

[0561]

[0562] The LCMS calculated value of C24H25N4O is 385.2 (M+H+), and the measured value is 384.1.

[0563]

[0564] N-((5-methoxy-1H-indol-3-yl)methyl)-1-methyl-1H-benzo[d]imidazol-2-amine (AJ2-88): synthesized according to general procedure 1, purified by biotage (hexane / ethyl acetate; 2:8) to give AJ2-87 (72 mg, 64%) as a yellow solid.

[0565]

[0566] The LCMS calculated value of C18H19N4O is 307.1 (M+H+), and the measured value is also 307.1.

[0567]

[0568] N-((9-ethyl-6-(4-methylpentyl)-9H-carbazole-3-yl)methyl)-1-methyl-1H-benzo[d]imidazol-2-amine (AJ2-89): synthesized according to general procedure 4 and subsequently by general procedure 1, purified by biotage (hexane / ethyl acetate; 4:6) to give AJ2-89 (68 mg, 72%) as a yellow solid.

[0569]

[0570]

[0571] The LCMS calculated value of C29H35N4 is 439.2 (M+H+), and the measured value is also 439.2.

[0572]

[0573] N-((1H-indol-5-yl)methyl)-3-(3-(but-3-yn-1-yl)-3H-diazadiazidin-3-yl)-N-(2-oxo-1,2,3,4-tetrahydroquinoline-3-yl)acrylamide (AJ2-90): synthesized according to general procedure 1 and general operation 2, and purified by PTLC (hexane / ethyl acetate; 5:5) to give AJ2-90 (22 mg, 46%) as a white solid.

[0574]

[0575] The LCMS calculated value of C26H26N5O2 is 440.2 (M+H+), and the measured value is 440.1.

[0576]

[0577] N-((1H-indol-5-yl)methyl)-N-(2-(azacycloheptane-1-yl)-2-phenylethyl)-3-(3-(but-3-yn-1-yl)-3H-diazapropidin-3-yl)acrylamide (AJ2-91): synthesized according to general procedure 1 and subsequently general procedure 2, purified by PTLC (hexane / ethyl acetate; 7:3) to give AJ2-91 (14 mg, 46%) as a colorless oil.

[0578]

[0579]

[0580] Note: Rotational isomers were observed. LCMS calculated value for C31H38N5O: 496.30 (M+H+), measured value: 496.1.

[0581]

[0582] 1-(2-(3-(but-3-yn-1-yl)-3H-diazapropidin-3-yl)ethyl)-N-((1-phenyl-1H-pyrazol-4-yl)methyl)-1H-benzo[d]imidazol-2-amine (AJ2-92): synthesized according to general procedure 1 and subsequently general procedure 5, purified by PTLC (hexane / ethyl acetate; 5:5) to give AJ2-92 (8 mg, 52%) as a yellow viscous liquid.

[0583]

[0584] The calculated LCMS (ESI) value of C24H24N7 is 410.2 (M+H+), and the measured value is 410.1.

[0585]

[0586] 1-(2-((benzo[d][1,3]m-dioxacyclopenten-5-ylmethyl)amino)-1H-benzo[d]imidazol-1-yl)but-1-one (AJ2-CP53): synthesized according to Scheme 1 and general procedure 4, and purified by PTLC (hexane / ethyl acetate 5:5) to give AJ2-CP53 (18 mg, 45%) as a grayish-white solid.

[0587]

[0588] The LCMS calculated value of C19H20N3O3 is 338.1 (M+H+), and the measured value is also 338.1.

[0589] Example 3. Fragment-based chemical proteomics approach for developing SLC inhibitors

[0590] Chemical probes offer a valuable method for directly querying the function and disease relevance of proteins, complementing genetic approaches by generating reversible and hierarchical gains or losses in protein activity and, in a variety of cases, novel functional outcomes. Chemical probes are typically discovered through high-throughput screening (HTS) of large chemical libraries (approximately 10⁶) against a desired target or phenotype. While HTS has been successfully implemented several times, this approach continues to face major technical hurdles limiting its versatility. First, they are typically performed using large libraries of structurally complex (MW 400 to 600 Da) compounds, which can be difficult to optimize due to their poor ligand efficiency.42 Additionally, such high-molecular-weight compound libraries have a large number of possible atomic combinations and therefore tend to inefficiently and incompletely explore the biologically relevant (“drugatable” chemical space throughout the human proteome.43,44 Fragment-based ligand discovery (FBLD) addresses some of the limitations of conventional HTS by determining smaller libraries (approximately 1000) of low-molecular-weight compounds (<300 Da) for binding to protein targets.42,43 By setting low molecular weight limits for compound screening, FBLD reduces the total number of possible atomic combinations by tens of orders of magnitude compared to the traditional molecular weight cutoff (around 500 Da) used for HTS.45 Fragment screening thus enables the exploration of a larger portion of the small molecule-protein interaction space with much smaller and more simplified compound libraries, often resulting in superior ligand efficiency compared to HTS hits.43 Fragment screening typically has higher hit rates than HTS; however, due to the low affinity of these hits, FBLD has so far been limited to studies of purified protein targets where ligand-protein interactions can be characterized by biophysical methods such as NMR and X-rays.42

[0591] To address these limitations, a method combining fragment-based ligand discovery with chemical proteomics (called Fragment-Based Ligand Mapping in Cells (FbLMiC)) was developed. Figure 1 A)) is a powerful method for the global identification of ligandable proteins and their ligand-forming sites 46,47. In this method, small molecule probes, called fully functionalized fragment (FFF) probes, contain ( Figure 1 B to C): 1) A structurally minimized “constant” region with a photoactivated diaziridinium group and an alkyne handle, which together enable UV-induced covalent modification, detection, enrichment, and identification of compounds binding to protein targets; and 2) a “variable” recognition region composed of structurally diverse small molecule fragments (MW < 300 Da) to facilitate interactions with subsets of the proteome. The significant advantages of FbLMiC are: 1) capture and recognition of probe-protein interactions from living cells, preserving unstable interactions that can be disrupted by cell lysis; 2) FFF probes interact at functional protein sites (e.g., active sites, allosteric sites, and protein-protein interaction sites); 3) efficient enrichment and identification of low-abundance and low-affinity proteins through covalent capture; and 4) fragments can be optimized for higher-affinity ligands through FbLMiC-guided medicinal chemistry. The platform boasts unprecedented proteomics coverage with an ever-growing ligand-forming profile of over 4,000 human proteins, including proteins that do not fall into traditional "drug-forming" categories (such as adaptors and transcription factors), and the vast majority of which currently lack chemical probes. Figure 1 C).

[0592] Using the FbLMiC platform, over 100 SLC-FFF interactions have been identified directly in human cell lines and primary immune cells. Furthermore, these FFF-SLC interactions have been demonstrated to serve as a starting point for SLC inhibitor development.46 For example, a coumarin-based FFF (FFF 3) that highly binds to the acylcarnitine transporter SLC25A20 was recently identified.46 A first-in-class inhibitor for SLC25A20 (CP22) was developed using FbLMiC-assisted medicinal chemistry and used to characterize SLC25A20 function in the context of fatty acid metabolism. The combination of FFF 3, structurally similar but non-SLC25A20-binding CP26, and CP22 demonstrated that: 1) the pharmacological inhibitor binds to the intermembranous side of the transport domain of SLC25A20 (…). Figure 2C ); 2) Blockage of SLC25A20 transport leads to the accumulation of long-chain acylcarnitine (>C14) ( Figure 2D(3) These indicate that these are major SLC25A20 substrates; and (4) blocking SLC25A20 transport inhibits fatty acid oxidation. As described in the next section, first-generation SLC15A4 inhibitors have been developed using FbLMiC. Here, it is suggested that these chemical probes be used to investigate the mechanisms by which SLC15A4 drives cytokine production in immune cells and to evaluate its potential as a therapeutic target for treating autoimmune diseases such as lupus.

[0593] Previous studies have identified SLC15A4 as playing a unique and crucial role in the production of IFN-I and other inflammatory cytokines in pDCs and in the pathogenesis of autoimmune diseases, thus elevating SLC15A4 to a potential therapeutic target for such conditions. However, to date, SLC15A4 remains uncrugable and no publicly available inhibitors exist. This application describes a feasible chemical proteomics strategy for developing a first-in-class inhibitor of SLC15A4 that blocks SLC15A4 transport and inhibits IFN-I production in human and mouse pDCs, intracellular target binding capacity, and structurally similar but inactive control compounds. Currently, the lead inhibitor can inhibit IFN-I production in primary human pDCs with an IC50–200 nM.

[0594] Chemical proteomics development of SLC15A4 chemical probes. To identify small molecular fragments that could serve as lead molecules for the development of SLC15A4 inhibitors, previously published datasets 46, 47 were searched using FbLMiC, and small (approximately 30) internal FFF libraries (20 and 200 mM) were screened in human peripheral blood mononuclear cells (PBMCs) by multiplex proteomics, as previously described 47. In short ( Figure 1 A) Freshly isolated PBMCs from healthy donor blood were treated with FFF for 30 minutes, exposed to UV irradiation to capture fragment-bound proteins, cleaved, conjugated to biotin-azide tags via copper(I)-catalyzed alkyne-azide cycloaddition (CuAAC, or “click chemistry”), and enriched with streptavidin-coated beads. The enriched proteins were digested with trypsin, and the resulting peptides were labeled with tandem mass spectrometry tags (TMT, for quantification), analyzed by LC-MS / MS / MS, and identified and quantified by their MS1 / MS2 and MS3 signals, respectively. Candidate targets were defined as proteins that were >5-fold enriched by hitting FFF compared to the control FFF (methyl fragment). Several fragments significantly enriched in endogenous SLC15A4 were identified. Figure 3ATo prioritize potential leads that could disrupt SLC15A4 function, their ability to modulate IFN-I production in TLR-stimulated human pDCs was examined. Briefly, human pDCs were isolated from PBMCs using a commercially available negative selection kit (Miltenyi), treated with CpG-A DNA (4 μg / mL) for 1 hour, followed by treatment with FFF (20 µM) and incubation overnight at 37°C. The next day, the supernatant was harvested and IFN-α levels were quantified by ELISA. FFF 5 was found to significantly inhibit IFN-I levels (Figure 4), while others showed moderate inhibition (not shown). Additionally, a structurally similar probe (FFF 6) was identified that neither binds to SLC15A4 nor inhibits IFN-I. "Non-clickable" forms of FFF 5 and FFF 6 were subsequently synthesized for use via a fluorescent gel-based assay. Figure 3B ) and MS-based experiments ( Figure 3C The two studies identified high stoichiometric ligand-protein interactions through a competition experiment against an excess of FFF 5 + / - "unclickable" competitors. Fourteen proteins were identified that were significantly enriched by FFF 5 (20 µM) and competed (>4-fold) with an excess of 5-comp, several of which were also competed against by the inactive control 6-comp. The most competitive target was SLC15A4, which was not competed against by the inactive 6-comp, indicating that it is a high stoichiometric target for both FFF 5 and 5-comp. Figure 2C To D). Furthermore, no evidence of interaction between FFF5 and SLC15A3 was observed in any proteomics studies. Subsequently, it was determined that FFF5 inhibits IFN-I production in a dose-dependent manner (IC50 approximately 1 mM). Figure 4A Furthermore, both FFF 5 and 5-comp, rather than the inactive analogue 6, inhibited the production of inflammatory cytokines in both human and mouse pDCs (isolated and described in target 1). Figure 4B To D).

[0595] Development of an NF-κB reporter gene assay for SLC15A4 transport. To assess whether FFF 5 and related compounds inhibit SLC15A4 transport, reporter cell lines based on SLC15A4 transport were generated. The precise substrate scope of endolysosomal SLC15A4 has not been determined. However, several studies have shown that SLC15A4 is a transporter of bacterial-derived peptidoglycans (e.g., MDP and Tri-DAP, which are ligands for the immunosensors NOD1 and NOD2)27-29,48. Recently, it has been shown that disruption of the dileucine motif (DE)-XXXL-(L / I) or DXXLL of SLC15A349 and SLC15A450 can lead to successful targeting of the plasma membrane. Given that NOD transmits signals via the NFκB pathway, an NFκB reporter gene assay was developed as a strategy for measuring SLC15A4 transport in cells.

[0596] In short, wild-type (WT) human SLC15A4 and dileucine mutants (L14A, L15A, L318A, V319A) were cloned in-frame with mCherry using the (GGGGS)3 adapter in the pLPC lentiviral backbone. The lentiviral vector was packaged in psPAX2 and pMD2.G packaging plasmids and used to generate stable cell lines expressing SLC15A4 WT or membrane-transported SLC15A4 mutant A549 cells. Figure 5A Stable reporter cells were then generated from these SLC15A4 cell lines using a Promega dual luciferase reporter system (pGLA4.32[luc2P / NFκB-RE / Hygro]) containing five copies of the NF-κB response element. The production of luciferase following exposure to MDP or Tri-DAP NOD ligands was identified. Figure 6B To C), it should be exposed to triptolide (an NFκB inhibitor, Figure 5C It is inhibited by ML130 (NOD1 inhibitor, not shown) or GSK717 (NOD2 inhibitor, not shown). In addition, FFF5, but not FFF6, blocks MDP-induced luciferase, which together indicate that FFF5 blocks SLC15A4-mediated transport.

[0597] Preliminary structure-activity relationship (SAR) studies. Without prior structural or technological support, a robust and simple synthetic strategy was developed that provides a pathway to obtain numerous and diverse scaffolds from readily available starting materials in 2 to 3 simple synthetic steps, enabling rapid synthetic exploration of chemical characterizations that can enhance SLC15A4 inhibitory activity and (if desired) improve PK properties for in vivo studies (Objective 2). To this end, the 5-comp was divided into three main chemical regions: benzimidazole-purple, butyryl-red, and benzo[a]dioxane-pentene / aromatic-green. Figure 6A ), and in the preliminary study, a simple synthetic route was used ( Figure 6A Eighteen analogues of 5-comp (7 to 24) were synthesized. Figure 6B In short, readily available aromatic aldehydes (green) are coupled with benzimidazole (Int-1A, top route) or other aromatic amines (Int-1B, bottom route) using standard reductive amination conditions to provide intermediates Int-A2 / Int-B2, which can then be diversified at the N1 position of benzimidazole by treatment with a wide variety of acyl chlorides or coupling with acids (3) or by alkylation. The cytotoxicity of the analogues in primary human pDCs was first examined at 10 μM, revealing no signs of metastable toxicity. Next, the following abilities of 7 to 24 were examined: 1) inhibition of IFN-I production in TLR7 (CpG)-stimulated human pDCs; and 2) blocking SLC15A4-mediated MDP transport (… Figure 6D Almost all substitutions of the benzimidazole ring (21 to 24) destroyed the activity, while multiple substitutions at the butyryl (red, 16 to 20) and benzo[a]dioxane (green, 7 to 15) positions were more tolerant. Specifically, 5-bromoindole analog 8 was found to be the most effective in both assays, with an IFN-I inhibition IC50 of 190 nM. Figure 6E The activity was approximately 5-fold higher than that of 5-comp. Several inactive analogues (13-17, 20-24) were also identified, representing other valuable control compounds for functional studies. Notably, a strong correlation was found between the activities of each compound in the two assays; that is, analogues that inhibit IFN-I production also blocked MDP transport to a similar degree. Figure 6D The consistency between the assays suggests that the interaction between the inhibitor and SLC15A4 similarly affects the mechanism of both activities, and is therefore consistent with the hypothesis that the transport function is mechanistically related to TLR signaling in the subsequent pDC.

[0598] By incorporating via reference

[0599] All U.S. patents and U.S. patent application publications cited in this article are incorporated herein by reference.

[0600] Equivalent scheme

[0601] Those skilled in the art will recognize, or can determine, many equivalents of the specific embodiments of the present disclosure described herein using only conventional experiments. These equivalents are covered in the appended claims.

[0602] References

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Claims

1. A compound of formula (I) or (II), or a pharmaceutically acceptable salt thereof: ， in X is NR 4 Or S; R 1 It is H or -C(O)-C 1-10 alkyl; R 2 It is an optional substituted C6-C 12 Aryl or 3 to 12-membered heterocyclic group, optionally substituted with a cyano, halogen, hydroxyl, or unsubstituted C. 1-10 Substitution of one or more of the alkyl groups; or R 2 yes 、 ； R 3 It is halogen; R 4 It is H, or unsubstituted -C 1-10 Alkyl, -C(O)-C 1-10 Alkyl, -C(O)-C 3-10 cycloalkyl, or -S(O)2-C 1-10 alkyl or ;as well as n is 0, 1, 2, 3 or 4.

2. The compound of claim 1, wherein the compound is a compound of formula (I).

3. The compound of claim 1, wherein the compound is a compound of formula (II).

4. The compound according to any one of claims 1 to 3, wherein R 1 It's H.

5. The compound according to any one of claims 1 to 3, wherein R 1 It is -C(O)-C 1-10 alkyl.

6. The compound of claim 5, wherein R 1 It is -C(O)-C3H7.

7. The compound of claim 1, wherein R 2 It is an unsubstituted 3 to 12-membered heterocyclic group.

8. The compound of claim 1, wherein R 2 It is a substituted 3 to 12-membered heterocyclic group.

9. The compound of claim 1, wherein the heterocyclic group is monocyclic.

10. The compound of claim 1, wherein the heterocyclic group is bicyclic.

11. The compound of claim 1, wherein the heterocyclic group is tricyclic.

12. The compound of claim 1, wherein the 3- to 12-membered heterocyclic group is aromatic.

13. The compound of claim 1, wherein the 3- to 12-membered heterocyclic group is non-aromatic.

14. The compound of claim 1, wherein R 2 It is unreplaced C6-C 12 Aryl.

15. The compound of claim 1, wherein R 2 It is a substituted C6-C 12 Aryl.

16. The compound of claim 1, wherein R 2 yes: 。 17. The compound of claim 1, wherein R 2 yes: 。 18. The compound of claim 1, wherein R 2 yes 。 19. The compound of claim 1, wherein R 2 yes 。 20. The compound of claim 1, wherein R 3 It is F.

21. The compound of claim 1, wherein R 3 It is Cl.

22. The compound of claim 1, wherein R 3 It is Br.

23. The compound of claim 1, wherein R 4 It's H.

24. The compound of claim 1, wherein R 4 It is unreplaced -C 1-10 alkyl.

25. The compound of claim 24, wherein R 4 It is a methyl group.

26. The compound of claim 1, wherein R 4 It is unsubstituted -C(O)-C 1-10 alkyl.

27. The compound of claim 26, wherein R 4 yes 。 28. The compound of claim 26, wherein R 4 It is -C(O)-C3H7.

29. The compound of claim 1, wherein R 4 It is unsubstituted -C(O)-C 3-10 Cycloalkyl.

30. The compound of claim 29, wherein R 4 It is -C(O)-cyclopropyl.

31. The compound of claim 29, wherein R 4 It is -C(O)-cyclohexyl.

32. The compound of claim 1, wherein R 4 It is unsubstituted -S(O)2-C 1-10 alkyl.

33. The compound of claim 32, wherein R 4 It is -S(O)2-C3H7.

34. The compound of claim 1, wherein R 4 yes 。 35. The compound of claim 1, wherein n is 0.

36. The compound of claim 1, wherein n is 1.

37. The compound of claim 1, wherein n is 2.

38. The compound of claim 1, wherein n is 3.

39. The compound of claim 1, wherein n is 4.

40. A compound, wherein the compound is one of the following or a pharmaceutically acceptable salt thereof: 1-(2-(((1H-indol-5-yl)methyl)amino)-1H-benzis[d]imidazol-1-yl)but-1-one; N-((1H-indol-5-yl)methyl)-N-(benzo[d]thiazo-2-yl)butyramide; N-((5-bromo-1H-indol-3-yl)methyl)-1H-benzo[d]imidazol-2-amine; 1-(2-(((5-bromo-1H-indol-3-yl)methyl)amino)-1H-benzo[d]imidazol-1-yl)but-1-one; 1-(2-((isoquinoline-5-ylmethyl)amino)-1H-benzo[d]imidazol-1-yl)but-1-one; 1-(2-(((1H-pyrrolo[2,3-b]pyridin-3-yl)methyl)amino)-1H-benzo[d]imidazol-1-yl)but-1-one; 1-(2-((pyrazolo[1,5-a]pyridin-5-ylmethyl)amino)-1H-benzo[d]imidazol-1-yl)but-1-one; 1-(2-((3,4-dimethoxybenzyl)amino)-1H-benzo[d]imidazol-1-yl)but-1-one; 1-(2-(((1-benzyl-1H-indol-5-yl)methyl)amino)-1H-benzo[d]imidazol-1-yl)but-1-one; 1-(2-(((1H-benzo[d]imidazol-5-yl)methyl)amino)-1H-benzo[d]imidazol-1-yl)but-1-one; (2-(((1H-indol-5-yl)methyl)amino)-1H-benzo[d]imidazol-1-yl)(cyclopropyl) ketone; N-((1H-indol-5-yl)methyl)-1-(propylsulfonyl)-1H-benzo[d]imidazol-2-amine; 1-(2-(((1H-indol-5-yl)methyl)amino)-1H-benzi[d]imidazol-1-yl)-3,5,5-trimethylhex-1-one; 1-(2-(((1H-indol-5-yl)methyl)amino)-1H-benzo[d]imidazol-1-yl)-3-cyclopentylprop-1-one; (2-(((1H-indol-5-yl)methyl)amino)-1H-benzo[d]imidazol-1-yl)(cyclohexyl) ketone; 1-(2-(((1H-indol-5-yl)methyl)amino)-1H-benzimidazol-1-yl)-3-(3-(but-3-yn-1-yl)-3H-diazadiazin-3-yl)prop-1-one; 1-(2-(((6-methoxypyridin-3-yl)methyl)amino)-1H-benzo[d]imidazol-1-yl)but-1-one; 1-(2-(((5-fluoro-1H-indol-3-yl)methyl)amino)-1H-benzis[d]imidazol-1-yl)but-1-one; 1-(2-((dibenzo[b,d]furan-4-ylmethyl)amino)-1H-benzo[d]imidazol-1-yl)but-1-one; 1-(2-(((5-(4-fluorophenyl)-1H-pyrazol-4-yl)methyl)amino)-1H-benzo[d]imidazol-1-yl)but-1-one; 1-(4-(((1-Butyryl-1H-benzo[d]imidazol-2-yl)amino)methyl)-5-(4-fluorophenyl)-1H-pyrazol-1-yl)but-1-one; N-((1H-benzo[d]imidazol-2-yl)methyl)-N-((1H-indol-5-yl)methyl)butyramide; 1-(2-(((1H-indol-5-yl)methyl)amino)-5-fluoro-1H-benzo[d]imidazol-1-yl)but-1-one; N-((9-ethyl-9H-carbazol-3-yl)methyl)-1H-benzo[d]imidazol-2-amine; 1-(2-(((9-ethyl-9H-carbazol-3-yl)methyl)amino)-1H-benzo[d]imidazol-1-yl)but-1-one; 1-(2-(((1-phenyl-1H-pyrazol-4-yl)methyl)amino)-1H-benzo[d]imidazol-1-yl)but-1-one; N-((5-bromo-1H-indol-3-yl)methyl)-1-methyl-1H-benzo[d]imidazol-2-amine; N-((1H-indol-5-yl)methyl)-1-methyl-1H-benzo[d]imidazol-2-amine; N-((9-ethyl-9H-carbazol-3-yl)methyl)-1-methyl-1H-benzo[d]imidazol-2-amine; N-((9-ethyl-9H-carbazol-3-yl)methyl)-N-(1-methyl-1H-benzo[d]imidazol-2-yl)butyramide; 1-(2-(3-(but-3-yn-1-yl)-3H-diazadiazin-3-yl)ethyl)-N-((9-ethyl-9H-carbazol-3-yl)methyl)-1H-benzo[d]imidazol-2-amine; N-((1,6,7,8-tetrahydrocyclopentano[g]indol-3-yl)methyl)-1H-benzo[d]imidazol-2-amine; 1-(2-(((1,6,7,8-tetrahydrocyclopentano[g]indol-3-yl)methyl)amino)-1H-benzo[d]imidazol-1-yl)but-1-one; 3-(((1H-benzo[d]imidazol-2-yl)amino)methyl)-1H-indol-6-carboxynitrile; N-((5-chloro-1H-indol-3-yl)methyl)-1H-benzo[d]imidazol-2-amine; 9-Ethyl-6-(((1-Methyl-1H-benzo[d]imidazol-2-yl)amino)methyl)-9H-carbazole-3-carboxynitrile; N-((9-ethyl-3,3-dimethyl-2,3,4,9-tetrahydro-1H-carbazol-6-yl)methyl)-1-methyl-1H-benzo[d]imidazol-2-amine; N-((9-ethyl-3,3-dimethyl-2,3,4,9-tetrahydro-1H-carbazol-6-yl)methyl)-1H-benzo[d]imidazol-2-amine; 1-Butyl-N-((9-ethyl-3,3-dimethyl-2,3,4,9-tetrahydro-1H-carbazol-6-yl)methyl)-1H-benzo[d]imidazol-2-amine; 1-Butyl-N-((9-ethyl-9H-carbazol-3-yl)methyl)-1H-benzo[d]imidazol-2-amine; N-((1H-indol-5-yl)methyl)-1-butyl-1H-benzo[d]imidazol-2-amine; 1-Butyl-N-(dibenzo[b,d]furan-4-ylmethyl)-1H-benzo[d]imidazol-2-amine; 1-Butyl-N-((6-methoxypyridin-3-yl)methyl)-1H-benzo[d]imidazol-2-amine; N-(benzo[d][1,3]m-dioxacyclopenten-5-ylmethyl)-1-butyl-1H-benzo[d]imidazol-2-amine; N-((5-bromo-1H-indol-3-yl)methyl)-1-butyl-1H-benzo[d]imidazol-2-amine; N-((9-ethyl-3-(trifluoromethyl)-2,3,4,9-tetrahydro-1H-carbazol-6-yl)methyl)-1-methyl-1H-benzo[d]imidazol-2-amine; N-((9-ethyl-3-(trifluoromethyl)-2,3,4,9-tetrahydro-1H-carbazol-6-yl)methyl)-1H-benzo[d]imidazol-2-amine; 1-Butyl-N-((9-ethyl-3-(trifluoromethyl)-2,3,4,9-tetrahydro-1H-carbazol-6-yl)methyl)-1H-benzo[d]imidazol-2-amine; (R)-1-(1H-benzo[d]imidazol-2-yl)-N-((9-ethyl-9H-carbazol-3-yl)methyl)ethane-1-amine; 3-(((1H-benzo[d]imidazol-2-yl)amino)methyl)-1H-indole-5-carboxylic acid methyl ester; 3-(((1H-benzo[d]imidazol-2-yl)amino)methyl)-1H-indole-5-carboxylic acid; N-((1H-benzo[g]indol-3-yl)methyl)-1H-benzo[d]imidazol-2-amine; N-((1H-benzo[g]indol-3-yl)methyl)-1-methyl-1H-benzo[d]imidazol-2-amine; 1-(2-(3-(but-3-yn-1-yl)-3H-diazadiazino-3-yl)ethyl)-N-(3,4-dimethoxybenzyl)-1H benzo[d]imidazol-2-amine; N-((7-ethyl-7H-benzo[c]carbazole-10-yl)methyl)-1H-benzo[d]imidazol-2-amine; N-((7-ethyl-7H-benzo[c]carbazole-10-yl)methyl)-1-methyl-1H-benzo[d]imidazol-2-amine; N-((9-(4-(tert-butyl)phenyl)-9H-carbazol-3-yl)methyl)-1-methyl-1H-benzo[d]imidazol-2-amine; N-((6-(3,5-dimethylisocyanate) (-4-yl)-9-ethyl-9H-carbazol-3-yl)methyl)-1H-benzo[d]imidazol-2-amine; N-((6-(3,5-dimethylisocyanate) (-4-yl)-9-ethyl-9H-carbazol-3-yl)methyl)-1-methyl-1H-benzo[d]imidazol-2-amine; N-((6-bromo-9-ethyl-9H-carbazol-3-yl)methyl)-1-methyl-1H-benzo[d]imidazol-2-amine; N-((6-(benzo[d][1,3]m-dioxacyclopenten-5-yl)-9-ethyl-9H-carbazol-3-yl)methyl)-1-methyl-1H-benzo[d]imidazol-2-amine; 3-(9-ethyl-6-(((1-methyl-1H-benzo[d]imidazol-2-yl)amino)methyl)-9H-carbazole-3-yl)phenol; N-((9-ethyl-6-methoxy-9H-carbazol-3-yl)methyl)-1-methyl-1H-benzo[d]imidazol-2-amine; N-((5-methoxy-1H-indol-3-yl)methyl)-1-methyl-1H-benzo[d]imidazol-2-amine; N-((9-ethyl-6-(4-methylpentyl)-9H-carbazol-3-yl)methyl)-1-methyl-1H-benzo[d]imidazol-2-amine; 1-(2-(3-(but-3-yn-1-yl)-3H-diazapropidin-3-yl)ethyl)-N-((1-phenyl-1H-pyrazol-4-yl)methyl)-1H-benzo[d]imidazol-2-amine; 1-(2-((benzo[d][1,3]m-dioxacyclopenten-5-ylmethyl)amino)-1H-benzo[d]imidazol-1-yl)but-1-one N-((1H-indol-5-yl)methyl)-N-(2-oxo-1,2,3,4-tetrahydroquinoline-3-yl)butyramide; N-((1H-indol-5-yl)methyl)-N-(2,3-dihydro-1H-inden-2-yl)butyramide; 3-(3-(but-3-yn-1-yl)-3H-diazadiazin-3-yl)-N-((9-ethyl-9H-carbazol-3-yl)methyl)-N-(1-methyl-1H-benzo[d]imidazol-2-yl)acrylamide; N-((5-bromo-1H-indol-3-yl)methyl)-3-(3-(but-3-yn-1-yl)-3H-diazolicid-3-yl)-N-(1-methyl-1H-benzo[d]imidazol-2-yl)acrylamide; N-((9-ethyl-9H-carbazole-3-yl)methyl)quinoline-3-amine.

41. A pharmaceutical composition comprising the compound of claim 1 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

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