Compounds and methods for the treatment and prevention of fibrous disease conditions and cancer

Abstract

The present invention provides compounds for treating and preventing fibrotic disease conditions and / or cancer. [Solution] Compounds, pharmaceutical compositions, and methods are provided for reprogramming M2-like macrophages into M1-like macrophages and reversing the anti-fibrotic to pro-fibrotic shift observed during the course of fibrotic diseases and certain cancers. The compounds comprise immunomodulators that target cellular pattern recognition receptors and are specific to target cells through the incorporation of a targeting moiety (e.g., folic acid or a functional fragment or analog thereof). Releaseable and / or non-releaseable linkers may be included and manipulated to facilitate optimal delivery of the immunomodulator. The compounds and compositions may be employed in one or more therapeutic methods for fibrotic diseases and / or cancers.

Description

[Technical Field]

[0001] Priority This application is a) U.S. Provisional Patent Application No. 62 / 871,68 filed on 8 July 2019. No. 6, and b) U.S. Provisional Patent Application No. 62 / 872,14 filed on 9 July 2019 In relation to Article 6, the benefit of those priority rights is asserted. The contents of the aforementioned application are, by reference, in their entirety. The body is incorporated herein by reference.

[0002] This disclosure includes a targeting portion that reprograms M2 macrophages into M1 macrophages. Using one or more Gram-forming compounds, fibrous disease conditions and / or cancer are used to treat them. This invention relates to compounds, pharmaceutical compositions, and methods for preventing and preventing [unclear / unclear]. [Background technology]

[0003] Despite the severity of many fibrotic diseases, such as idiopathic pulmonary fibrosis (IPF), treatment is effective. There are few options for making it work, and almost everything that has been available conventionally has an underlying condition. It is designed not to cure, but to alleviate symptoms and slow progression. For example, I In PF, oxygen therapy can improve comfort and lifestyle, but it can also worsen disease progression. It has little effect against. Similarly, two FDA-approved drugs, pirfenidone Nintedanib may slow disease progression, but it does not restore existing fibrosis. It is not possible to stop the production of further fibrotic diseases such as IPF. Given the high probability of death associated with vascular disease, slowing disease progression and probably There is a great need to identify new strategies to halt the progress.

[0004] Furthermore, cancer is treated with very rare drugs such as mitomycin, paclitaxel, and camptothecin. It is treated with chemotherapy using powerful drugs. In many cases, these chemotherapy agents are administered in large doses. It shows a reactive effect, and tumor inhibition is proportional to the drug dose. Therefore, to treat neoplasms Aggressive administration regimens are used for this purpose, but high-dose chemotherapy has selectivity for cancer cells. This is hindered by its low efficacy and toxicity to normal cells. The lack of tumor specificity is a problem with conventional chemicals. It is one of many hurdles that need to be overcome through therapy.

[0005] Despite the clear need for the prevention and treatment of both fibrous diseases and cancer, These conditions are those for which there are currently no effective treatment options available to treat them. Therefore, it remains a significant cause of death and / or suffering worldwide. Furthermore, drugs or other If therapies are available, such treatments are for the target fibrous and / or cancer. Due to its poor selectivity for cells, it usually carries a risk of systemic toxicity in the underlying target. Yes, a very powerful drug will be used. What is needed is activated M2 type (alternatively activated Pro-fibrosis and / or pro-proliferative factors initiated by (sexualized) macrophages Not only does it disrupt the cycle, but it also affects problematic cells (cancer cells or fibrous cells). It does this with very high specificity to other cells (regardless of whether they are other cells experiencing the same condition). This is also a possible treatment. [Overview of the Initiative] [Problems that the invention aims to solve]

[0006] In some cases, activated M2 phenotype macrophages activate fibroblasts, for example. It secretes fibrosis-promoting cytokines that synthesize collagen and other extracellular matrix proteins. By doing so, they play a role in fibrous diseases. In certain cases, these macrophages Similarly, this triggers the release of growth factors that are problematic in subjects experiencing cancer. For example, such growth factors can promote the growth of cancerous tumors. Furthermore, in some cases, Macrophages release immunosuppressive cytokines (for example, concurrently). However Macrophages promote the establishment and growth of fibrous diseases and / or cancer. It can play an important role.

[0007] Idiopathic pulmonary fibrosis (IPF) is one such fibrotic disease, for example, collagen It is an interstitial lung disease resulting from excessive deposition of cellulose. In some cases, this type of fibrosis affects the lungs. This leads to progressive rigidification of the lungs, and in some cases, loss of the lungs' ability to mediate gas exchange. This progressive decline in lung capacity has resulted in a median survival time after IPF diagnosis being only 2 years. It is estimated to last 0.5 to 5 years. In some cases, serious related pathological conditions may occur (e.g., chronic hypoxia). Symptoms include fatigue, weight loss, muscle and joint pain, persistent cough, and loss of mobility. It continues to increase during the later stages of pathology. In the United States, approximately 40,000 new cases of IPF are diagnosed annually. They are cut off, and most of them die.

[0008] In some cases, activity originates from macrophages or peripheral blood monocytes present in the tissue. Macrophages are characterized by the chemokine (CC motif) ligand 18 (CCL18), and their characteristics Conversion growth factor-β1 (TGFβ1) and / or platelet-derived growth factor (PDGF) It induces fibroblast activation through secretion. In some cases, this activation leads to fibroblasts It promotes the secretion of collagen, which helps to prevent the progression of fibrous diseases and related cancers. In the later stages of many fibrous diseases, activated macrophages and myofibroblasts interact with each other. Cross-stimulation can occur, leading to the propagation of fibrosis throughout the lungs or other relevant parts of the body. This creates a vicious cycle that ensures certainty.

[0009] Similar conditions are observed in other fibrous diseases. Cancer also promotes the growth of malignant tumors. (For example, by growth factors secreted by activated macrophages), and / Or promote collagen formation in cancerous tumors (e.g., downstream fibrous collagen). Through its production, it blocks the drug permeability of cancer, making it more difficult to treat. It may be accompanied by a similar immune response (which can lead to tumors). [Means for solving the problem]

[0010] In some embodiments, compounds represented by formula QLT are provided herein. In some embodiments, Q is a radical of a folate receptor-binding ligand. In one embodiment, L is a linker. In some embodiments, T is a toll-like It is a radical of a receptor (TLR) agonist. In some embodiments, QLT is It is a pharmaceutically acceptable salt.

[0011] In some embodiments, the linker is a non-discharge linker. So, the non-release linker is given by the formula, [ka] It is represented as follows.

[0012] In some embodiments, n is 1 to 30. In some embodiments, n is 1 It is ~24. In some embodiments, n is 1 to 12. In some embodiments, n is 1 to 3. In some embodiments, n is 12. Morphologically, n is 3.

[0013] In some embodiments, w is 0 to 5. It is 2. In some embodiments, w is 1.

[0014] In some embodiments, TLR agonists are Toll-like receptor 7 (TLR7) agonists. It is a agonist. In some embodiments, the radical of the TLR agonist is expressed by the following equation X. It has a structure that can be represented as follows. [ka]

[0015] In some embodiments, R1 is -NH2 or -NH-R 1X That is. Several In the embodiment, R2 is H, alkyl, alkenyl, alkynyl, alicyclic lic), aryl, biaryl, heteroaryl, -NH-R 2X , -OR 2X ,- SR 2X , [ka] In some embodiments, R 1X , R 2X , and R 2Y Each of them is independent of water is selected from the group consisting of element (H), alkyl, alkenyl, alkynyl, alicyclic, aryl, biaryl, and heteroaryl. In some embodiments,

Chemical Structure

Chemical Structure

Chemical Structure

[0016] In some embodiments, the radical of the TLR agonist has a structure represented by Formula XX. [ka]

[0017] In some embodiments, R1 is -NH2 or -NH-R 1X That is. Several In the embodiment, R2 is H, alkyl, alkenyl, alkynyl, alicyclic, aryl, and Aryl, heteroaryl, -NH-R 2X , -OR 2X , -SR 2X , [ka] In some embodiments, R 1X , R 2X , and R 2Y Each of them is independent of H alkyl, alkenyl, alkynyl, alicyclic, aryl, biaryl, and hetero Selected from the group consisting of reels. In some embodiments, [ka] This is a 3- to 10-membered nitrogen-containing non-aromatic monocyclic or bicyclic heterocycle. Several embodiments Then, X is CH, CR2, or N. In some embodiments, R1 is -NH 2 or -NH-R 1X R2 is H, alkyl, alkenyl, alkynyl, alicyclic Formula, aryl, biaryl, heteroaryl, -NH-R 2X , -OR 2X , -SR 2X , [ka] And R 1X , R 2X , and R 2Y Each of them independently consists of H, alkyl, alkenyl, Select from the group consisting of alkynyl, alicyclic, aryl, biaryl, and heteroaryl. And so, [ka] X is a 3-10 member nitrogen-containing non-aromatic monocyclic or bicyclic heterocycle, where X is CH, CR2 , or N.

[0018] In some embodiments, the radical of the TLR7 agonist is represented by formula XXX. It has a structure that [ka]

[0019] In some embodiments, the compound is a targeted moiety and an immunomodulator or its pharmaceutically acceptable Linker L between acceptable salts n It further includes Linker L n is a TLR7 agonist It is configured to avoid the release of the free form, where n is an integer less than or equal to 50. In this embodiment, linker L n This is polyethylene glycol (PEG) or a PEG derivative. It includes n, where n is an integer selected from the range of 1 to 32, and the folate receptor binding ligand Dical is a folate receptor β (FBβ) binding ligand.

[0020] In some embodiments, the compound has the structure shown below. [ka]

[0021] In some embodiments, the compound has the structure shown below. [ka]

[0022] In some embodiments, the compound has the structure shown below. [ka]

[0023] In some embodiments, the compound has the structure shown below. [ka]

[0024] In some embodiments, a pharmaceutical composition comprising one or more of the compounds of this disclosure is described herein. Provided in the book, the TLR7 agonist has a structure represented by formula XX.

[0025] In certain cases, the method of treating subjects suffering from fibrotic disease or cancer is described in this specification. Provided in the book, this method involves targeting cells with immunomodulatory substances that react with TLR7, 8, or 9 Contact with at least one compound containing an agonist of the compound described herein. This includes causing someone to do something.

[0026] In some embodiments, folate ligans attached to the TLR agonist via a linker are used. A compound comprising a functional fragment or analog thereof, having the following formula TLR Agonists or pharmaceutically acceptable salts thereof are provided herein. [ka]

[0027] In some embodiments, R 1 is an amine group, R 2 It is a single bond -NH-, R 3 is H, alkyl, hydroxyl group, or any other substituent thereof, and X is CH 2, NH, oxygen (O), or sulfur (S), and the linker is R 1 、 R 2 , or R 3 to It adheres to the surface.

[0028] In some embodiments, a pharmaceutical comprising any one compound of any of the formulas provided herein is used. A composition is provided, and the linker described above includes a PEG linker or a PEG derivative linker. Mi, R 3 It is a non-release linker attached in or R 1 、 R 2 , or R 3 It is a releaseable linker attached thereto.

[0029] In some embodiments, pharmaceutically acceptable salts are hydrobromide, citrate, and triglycerides. Fluoroacetate, ascorbic acid, hydrochloride, tartrate, triflate, maleate, me Selected from silates, formates, acetates, or fumarates.

[0030] In some embodiments of this specification, cells are linked to a folate ligand or An immunomodulatory substance or a pharmaceutically acceptable salt thereof attached to the functional fragment or analogue thereof. At least one compound containing (for example, provided by the formula provided herein) A method is provided for preventing or treating a fibrous disease condition, which includes contact with a compound of the same name. Therefore, the above-mentioned immunomodulators or their pharmaceutically acceptable salts target pattern recognition receptors. In some embodiments, the cells are experiencing or have experienced a fibrous disease state. The target cells include those that are at risk of being exposed to at least one compound. , further comprising administering or applying a therapeutically effective amount of at least one compound to the target. In some embodiments, the subjects are patients experiencing IPF, and at least one The compound is administered intravenously, intramuscularly, intraperitoneally, topically, or by inhalation to the target. In some embodiments, the fibrous disease state is IPF or the liver, skin, bladder, heart. This includes fibrotic diseases of the organs, pancreas, prostate, or kidneys.

[0031] In some embodiments, the method involves obtaining a sample from a subject, or obtaining a sample This involves quantifying the expression level of one or more biomarkers in the sample, Each of one or more biomarkers is CCL18, arginase 1 (Arg1), and Matri Metalloproteinase 9 (MMP9), metalloproteinase 3 (TIMP3), Centerleukin 1β (IL-1β), hydroxyproline, collagen, PDGF, TG Fβ, folate receptor β (FRβ), tumor necrosis α (TNFα), interferon gamma (IFN-γ), mannose receptor (CD206), cluster of differentiation antigens (IFN-γ), mannose receptor (CD206), differentiation) 163 (CD163), differentiation antigen group 86 (CD86) ), interleukin-6 (IL-6), chemokine-10 (CXCL10), and immunoglobulins Selected from the group consisting of feron (IFNα), to quantify and 1 in the sample The expression level of each of the more than one biomarkers is compared to the expression level of such biomarkers in the control group. Compare with current levels, and CCL18, Arg1, MMP9, TIMP3, IL-1β , PDGF, TGFβ, FRβ, CD206, CD163, hydroxyproline, or Is collagen upregulated compared to the control expression level? TNFα, IFN-γ, IL-6, CXCL10, IFNα, or CD86 are used as controls. When the expression level is downregulated or not expressed compared to the expression level of [the substance]. Administer a therapeutically effective dose of an unconjugated agonist or inhibitor to the subject. The act of administering, or the act of administering, further includes the act of administering.

[0032] In some embodiments, folic acid ligands or their functional fragments or analogs are FR It is specific to β and binds to FRβ on cells.

[0033] In some embodiments of this specification, immunomodulation targets cellular pattern recognition receptors. A compound is provided which comprises a targeted moiety attached to a substance or a pharmaceutically acceptable salt thereof. The targeted portion comprises a folic acid ligand or a functional fragment or analog thereof. [Brief explanation of the drawing]

[0034] The embodiments and other features, advantages, and aspects disclosed herein, as well as those related thereto The objectives to be achieved will be evident in light of the following detailed descriptions of various exemplary embodiments of this disclosure. This will likely be the case. Such detailed explanations can be better understood when considered together with the attached drawings. To be done: [Figure 1A]The chemical structures of exemplary compounds having a targeted moiety (folate receptor ligand) attached to an immunomodulator (Toll-like receptor 7 (TLR7) agonist radical) via a non-release linker (e.g., containing a polyethylene glycol (PEG) backbone) are shown. [Figure 1B] The chemical structure of an exemplary compound having a targeting moiety (folate receptor ligand) attached to an immunomodulator (TLR7 agonist radical) via a releaseable linker (e.g., containing a disulfide moiety in its skeleton), as well as an exemplary drug release mechanism, are shown. [Figure 2] This flowchart shows typical treatment methods for individuals who have or are at risk of having fibrous disease or cancer. [Figure 3A-3F] Graphs of various marker levels measured from human M2-type macrophages when exposed to exemplary free (untargeted) TLR7 agonists or exemplary targeted (e.g., those with folate receptor-binding ligands) TLR7 agonists at various concentrations for each compound are shown. The data shown in Figures 3A-3C support that administration of either an untargeted or targeted TLR7 agonist successfully reprogrammed M2-type macrophages to M1-type macrophages (i.e., downregulated M2-type pro-fibrotic macrophages), while the data shown in Figures 3D-3F support that administration of the tested compounds upregulated M1-type macrophages. Each value represents the mean ± SD for each group: #P<0.05, ##P<0.01, ##P<0.005, ####P<0.0001, and Dunnett's multiple comparison test for treated vs. untreated M2-type macrophages. [Figure 4A-4E]Representative graphical data of various marker levels measured from M2 macrophages incubated for 2 hours (Figures 4A–4E) or 46 hours (Figures 5A–5D) with exemplary free or targeted TLR7 agonists at various concentrations are shown. Figures 4A–4E and 5A–5D support the downregulation of the M2 type pro-fibrillation phenotype after administration of free and targeted TLR7 agonists. Each value represents the mean ± SD for each group: #P<0.05, ##P<0.01, ##P<0.005, ####P<0.0001, Dunnett's multiple comparison test for compound 1A and compound 1B treated groups versus M2 untreated groups in Figures 4A–5D. [Figures 5A-5D] Representative graphical data of various marker levels measured from M2 macrophages incubated for 2 hours (Figures 4A–4E) or 46 hours (Figures 5A–5D) with exemplary free or targeted TLR7 agonists at various concentrations are shown. Figures 4A–4E and 5A–5D support the downregulation of the M2 type pro-fibrillation phenotype after administration of free and targeted TLR7 agonists. Each value represents the mean ± SD for each group: #P<0.05, ##P<0.01, ##P<0.005, ####P<0.0001, Dunnett's multiple comparison test for compound 1A and compound 1B treated groups versus M2 untreated groups in Figures 4A–5D. [Figures 6A-6D] Graph data representing various marker levels measured from treated M2 macrophages with exemplary free and targeted TLR7 agonists at various concentrations are shown. These agonists were (i) incubated for 48 hours (Figures 6A and 6B), or (ii) incubated for 2 hours, then in fresh medium, and incubated for the remaining 46 hours (Figures 6C and 6D). Each value represents the mean ± SD for each group: #P<0.05, ##P<0.01, ##P<0.005, ####P<0.0001, compound 1A and compound 1B treated groups versus untreated M2 groups by Dunnett's multiple comparison test. [Figure 6E] Flow cytometry data supporting the finding that THP-1 (a human monocytic cell line derived from patients with acute monocytic leukemia)-induced macrophages were folate receptor beta (FRβ) positive (FRβ+) are shown. [Figure 6F] This demonstrates the stability of an exemplary targeted TLR7 agonist. [Figure 7A] The images show stained lungs taken from mice with bleomycin (BM)-induced experimental fibrosis and stained using anti-mouse FRβ antibody, with hematoxylin-eosin (H&E) staining performed on days 7, 14, and 21 after BM-induced lung injury. [Figure 7B] The quantification of FRβ staining in the panel shown in Figure 7A is presented. [Figure 7C-7D] Figure 7C shows FRβ immunohistochemistry (IHC) staining of lung tissue from a human with idiopathic pulmonary fibrosis (IPF) and healthy human lung tissue (Figure 7D). [Figure 7E] Images of mouse tissues / organs, taken from mice with or without BM-induced experimental fibrosis (phosphate-buffered saline (PBS) control), and imaged with folate receptor-targeted fluorescent dyes, are shown. [Figure 7F] This shows the fluorescence-activated cell sorting (FACS) analysis of mice with BM-induced experimental fibrosis. [Figure 8A] This shows the treatment plans for free and targeted TLR7 agonists in the BM model. [Figures 8B-8G] Figure 8A shows the levels of fibrosis-promoting markers (Figures 8B-8D) and anti-fibrosis markers (Figures 8E-8G) measured from mice treated with the BM model. [Figure 8H] Figure 8A shows the number of cells in bronchoalveolar lavage fluid (BALF) from mice treated with the BM model. [Figure 9A-9B] The survival curves (Figure 9A) and weight changes (Figure 9B) of pulmonary fibrosis mice treated with untargeted and targeted TLR7 drugs are shown. [Figure 10A] This shows the hydroxyproline content (μg / lung) in lung tissue as a measure of fibrosis. [Figure 10B-10C] Figure 9A shows lung tissue stained with H&E (Figure 10B) and Masson's trichrome (collagen) stain (Figure 10C). [Figures 11A-11B]The survival curves (Figure 11A) and weight changes (Figure 11B) of mice with pulmonary fibrosis treated with exemplary targeted TLR7 agonists are shown, with each value representing the mean ± SD for each group. [Figure 12] This disclosure demonstrates the dose-dependent effect of an exemplary targeted TLR7 agonist on the suppression of fibrosis in BM-induced mice. [Figure 12A] This graph shows data relating to the body weight of BM-induced mice over time. [Figure 12B] This specification shows the measurement of hydroxyproline content in lung tissue treated with various exemplary doses of conjugates (e.g., compound 1B). [Figure 12C] Images for histological analysis of lung tissue using various stains are shown. Each value represents the mean ± SD for each group, with *P<0.05, **P<0.005, and ***<0.0005, and Student's t-tests for saline vs. vehicle group and treatment group vs. vehicle group. [Figures 13A-13D] The following shows various marker levels measured from M2-like macrophages reprogrammed over 48 hours with exemplary targeted TLR7 agonists at various concentrations according to the method of this disclosure, and each value represents the mean ± SD for each group. [Figure 14A-14C] The following shows various marker levels measured from M2 macrophages reprogrammed according to the method of this disclosure using exemplary free and targeted TLR7 agonists at various concentrations. The values ​​shown in Figures 14A–14C represent the mean ± SD for each group; #P<0.05, ##P<0.005, ###P<0.0005, ####P<0.0001; Dunnett's multiple comparison test shows compound 3A, compound 3B treated, and compound 3C treated groups versus the untreated M2 group. [Figure 15] The levels of secreted chemokine (CC motif) ligand 18 (CCL18) protein in each group of 14A-14C cells after treatment with exemplary free TLR7 agonists are shown. [Figure 16] This paper presents the methodology of the BM mouse model. [Figures 17A-17B]The purity of exemplary targeted TLR7 agonists provided herein is shown. [Figures 18A-18F] Figure 16 shows data from the in vivo study methodology, including survival curves (Figure 18A), body weight changes (Figures 18B and 18D), cell concentration by BALF (Figure 18C), and hydroxyproline concentration (μgHP / leaf) in surviving mice (Figure 18E) and all mice (i.e., including both surviving mice and mice that died before day 21) (Figure 18F). [Figure 19] Both targeted and untargeted TLR7 agonists reprogram human monocyte-derived profibrosis macrophages to an antifibrosis phenotype (Figures 19A-19F). Mean ± SD. Statistical significance between groups was determined using unpaired two-sided t-tests (*P<0.05, **P<0.01, ***P<0.001, ****P<0.0001). [Figure 20] Figures 20A–20F show a comparison of plasma cytokine levels in healthy mice after treatment with compound 1A and compound 1B. Figure 20G shows the change in body weight after treatment of mice with the exemplary compounds provided herein, with body weight change as a measure of systemic toxicity during every-other-day administration (n=2), mean ± SD. Statistical significance between groups was compared using unpaired two-sided t-tests (*P<0.05, **P<0.01, ***P<0.001). [Figure 21] Figure 6 shows healthy and fibrous lungs stained with 4',6-diamido-2-phenylindole (DAPI) (nucleus, blue), anti-F4 / 80 (macrophage, red), and anti-mannose receptor (CD206).

[0035] This disclosure is susceptible to various modifications and alternative forms, but its exemplary embodiments are These are shown as examples in the drawings and described in detail herein. [Modes for carrying out the invention]

[0036] For the purpose of facilitating an understanding of the principles of this disclosure, embodiments shown in the drawings are used herein. , and describe it using a specific language. Nevertheless, the description of these embodiments It will be understood that no limitation on the scope is intended. On the contrary, The indication may be included in the spirit and scope of this application as defined by the attached claims. It is intended to cover substitutes, modifications, and equivalents. As mentioned above, this technology The compositions and compounds described herein may be illustrated and explained in one or more preferred embodiments. The objects and methods may include many different configurations, forms, materials, and accessories.

[0037] The following description contains numerous specific details in order to provide a complete understanding of this disclosure. Certain embodiments may be carried out without some or all of these specific details. This disclosure may not pertain to a specific biological system, a specific fibrous disease or cancer, or a specific Not limited to the organs or tissues of which, and naturally subject to change, the information provided herein is available. Please understand that this remains applicable in light of the available data.

[0038] The various technologies and mechanisms described herein describe connections or linkages between two components. There are combinations. Attached, linked, combined, connected, and their inflectional morphemes. Terms such as similar terms accompanied by the same meaning are either noted as differences or not otherwise clear from the context. They are used interchangeably as far as possible. These words and expressions do not necessarily imply a direct connection. Rather, it includes connections via intermediate components. The connection between the two components is various other Because one component can exist between the two components of interest, it is not necessarily direct and without obstacles. It should be noted that this does not necessarily mean connection. Therefore, unless otherwise specified. To the extent that it is true, a connection does not necessarily mean a direct and trouble-free connection.

[0039] Furthermore, whenever it is feasible and convenient, the same or similar parts may be depicted in the figures and descriptions. Similar reference numerals are used to refer to products or processes. Drawings are simplified rather than to exact scale. This is a simplified form. Naturally, this disclosure is presented in this manner solely for illustrative purposes. The principles and embodiments presented and described herein are specifically described herein. It is understood that this may apply to compounds and / or compositional components having a composition other than that of a single substance. In fact, the components of the compositions and compounds of this disclosure are formulated to facilitate their desired application. It is explicitly intended that it can be adjusted.

[0040] Unless otherwise defined, all technical and scientific terms used herein are chemical. It has the same meaning as generally understood by those skilled in the art of science and biology. Any method and materials similar or equivalent to those used in the practice or test of the subject matter of this application While other methods and materials may be used, preferred methods and materials are described herein. When used in this specification and the appended claims, the singular "a", "an " and "the" refer to multiple objects unless the content explicitly indicates otherwise. Therefore, for example, if the compound / composition is substituted with an alkyl or aryl, The compound / composition contains at least one alkyl and / or at least one aryl It can be arbitrarily replaced with. Furthermore, unless otherwise stated, the term "approximately" is used for percentages. Therefore, the range of the value is plus or minus 10%, and for the unit value, plus or minus 10%. It refers to a unit of minus 1.0; for example, "approximately 1.0" refers to a value in the range of 0.9 to 1.1. .

[0041] In certain embodiments, the compounds, compositions, and methods of this disclosure are used for the prevention of fibrotic diseases and Useful for treatment and / or therapy. In certain embodiments, the provided compounds and / or combinations The product is also useful for the prevention and / or treatment of cancer. In some embodiments, the present invention The compounds, compositions, and methods provided in this document target (for example, selectively) the innate immune system. By utilizing targeted strategies, the polarization of macrophages from M2 to M1 (polarizat) The ions are reprogrammed, for example, to utilize their anti-fibrotic properties.

[0042] In general, and without any intended limitations, the novel compounds, compositions, and The method targets the innate immune system and is designed to favor the anti-fibrotic properties of the M1 phenotype. This reprograms the polarization of macrophages from M2 type to M1 type. For example, less In one exemplary embodiment, such compounds and compositions are immunomodulators or Folate receptor-binding ligands or their radicals, conjugated to the pharmaceutically acceptable salt thereof. It includes a targeting portion that targets the folate receptor β (FRβ). As explained in detail below... These embodiments utilize the limited expression of FRβ to administer compounds systemically. It is directly localized to FRβ-expressing cells (e.g., cells of fibrous and / or cancerous tissues). As a result, immunomodulatory factor components are activated in bone marrow cells (e.g., M2-like macrophages). This allows for conversion to anti-fibrotic M1 polarization. This targeted design is for the immune system This effectively prevents systemic activation and thus avoids toxicity.

[0043] Further exemplary embodiments include a linker positioned between the targeting portion and the immunomodulator. This may include such linkers. These linkers may be releaseable or not releaseable. As described in the details, the compounds / compositions of this disclosure, including the releaseable linker, are administered If so, the time when the immunomodulator becomes active or around that time, the targeted portion and the immunomodulator The substances are released from each other. In addition or alternatively, the compounds / compositions of this disclosure release In embodiments including a non-extractable linker, when the targeted portion and the immunomodulator are administered, Under physiological conditions, it is not released immediately. In this way, it is taken up by target cells. After activation of the inoculum and / or immunomodulator, the components remain together.

[0044] Various embodiments of this disclosure, as well as supporting data on examples, will be described below. This will clarify the two main immune strategies found primarily in vertebrates: the innate immune system and the adaptive immune system. There is an innate immune response or nonspecific immune response, which is the first line of defense against non-autologous pathogens. It consists of physical, chemical, and cellular defenses. On the other hand, the adaptive immune system is the primary natural immune system. It is called upon to act against pathogens that evade or overcome immune defenses.

[0045] Inflammatory responses play an important role in immunity. When tissue is damaged, When a pathogen is detected, for example, an inflammatory response is initiated, and the immune system is activated. Innate immunity Immune cells of the system (i.e., neutrophils and eosinophils) transmit tissue damage via the vascular and lymphatic systems. They are first mobilized to the site of the wound, injury, or pathogen, followed by the mobilization of macrophages. It will be done.

[0046] Cells of the innate immune system are affected by specific proteins present in microbial pathogens or other non-self molecules. It is possible to express special pattern recognition receptors that sense and bind to sequences. When used, "pattern recognition receptors" refer to white blood cells—for example, at least macrophages. This refers to any immune receptor expressed on the membrane of the 217-217, including and including them, and activating the receptor. And ultimately the innate immune response (and in certain cases, the development of antigen-specific acquired immunity) It can bind to specific ligands that it brings about.

[0047] Examples of two classes of molecules that can bind to pattern recognition receptors include microorganisms. Pathogen-associated molecular patterns related to physical pathogens, and those released during cell damage or death. It contains damage-related molecular patterns associated with host cell components. Pattern recognition receptors Recognition of these protein sequences triggers the expression of specific genes whose products regulate the innate immune response. It can initiate an induced signaling pathway (for example, antigen-specific acquisition in some cases). (They direct the development of retonic immunity). Therefore, pattern recognition receptors are responsible for these signaling pathways. It mediates pathways that, in certain cases, positively or negatively control innate and even adaptive immune responses. It can be used for control.

[0048] Macrophages are a type of white blood cell known to eliminate pathogens through phagocytosis. It is a group in which specific differentiation occurs in response to the local tissue environment, They are broadly classified as having either an M1 or M2 phenotype. In some cases, macules Lophages are used to treat interferon-gamma (IFN-γ), lipopolysaccharide (LPS), and / or exposure to granulocyte-macrophage colony-stimulating factor (GM-CSF), It polarizes toward the M1 phenotype. In certain cases, the M1 phenotype is associated with a high level of inflammation. Production of induced cytokines (e.g., interleukin-1β (IL-1β), tumor necrosis factors) (TNF), Interleukin-12 (IL-12), Interleukin-18 (IL-1) 8) and / or resistance to pathogens (such as interleukin-23 (IL-23)). Ability to mediate, strong bactericidal properties, high production of reactive nitrogen and oxygen intermediates, and / or It is characterized by the enhancement of the T helper type 1 (Th1) response. In some cases, M1 polarization is spontaneous. It is associated with the "attack and killing" phase of the immune response. In certain cases, M1 polarization is , inhibit or prevent the initial establishment of infection and / or remove damaged tissue To be able to.

[0049] In a particular case, after the innate immune system has carried out this "attack and kill" phase, macrophage Phage can reprogram itself to become a healing system (i.e., M2 type), for example They may release growth factors to promote healing. Such growth factors are interleuk IL-4, Interleukin-10, Platelet-derived growth factor (PDG) F) Transforming growth factor-β1 (TGFβ), chemokine (CC motif) ) Ligand 18 (CCL18), and / or interleukin 13 (IL-13) It may include, but is not limited to, certain cytokines. In a particular example, this Exposure to such cytokines / growth factors alternatively activates the M2 macrophage phenotype. do.

[0050] In contrast to M1 macrophages, M2 macrophages may be associated with wound healing and tissue repair. As a result, M2 macrophages are involved in tissue remodeling, immunomodulation / suppression, and / or It is characterized by its involvement in tumor promotion. In certain cases, M2 macrophages induce cell proliferation. It produces polyamines and / or proline, which induce collagen production. The response is beneficial in healthy subjects, but the presence of M2 macrophages is associated with fibrous disease or Through immunosuppression and / or promotion of tumor growth and fibrosis in subjects affected by the disease It can have extremely harmful effects.

[0051] For example, fibrous conditions may begin with an unknown trauma or damage to the epithelium. In response to tissue damage resulting from this, chemokines and other factors are released to the damaged tissue. This can promote the infiltration of immune cells into the body (e.g., innate immune response), which can include, for example, M2-like cells. For example, monocytes and macrophages that release fibrosis-promoting cytokines are hypothesized to be the current form. This includes the chronic secretion of these cytokines, which then become endogenous and infiltrate tissues. Fibroblasts / fibrous cells are activated to become myofibroblasts, which then produce collagen and other fibers. They can secrete extracellular matrix proteins that can harden the surrounding tissue. In some cases, these M2 macules Clophages worsen the disease by promoting fibrosis. For example, idiopathic lung cancer. In patients with interstitial fibrosis (IPF), for example, M2 macrophages infiltrate the lungs, and fibrosis develops within them. This can promote and further degrade those functions. In some cases, the M2 phenotype is produced Growth factors and other cytokines promote the growth of cancerous tumors through similar pathways. .

[0052] Reprogramming of M2-like macrophages into M1-like macrophages In certain cancers and fibrous diseases, macrophages are associated with an anti-inflammatory (M2-like) phenotype. And it may be disproportionately biased. In certain cases, immunomodulators, for example, Activated bone marrow cells (e.g., M2-like macrophages) undergo anti-fibrotic M1 polarization (e.g., They produce little to no growth factors and / or related cytokines, for example, Converting (to slow down or eliminate the progression of the disease state) - for example, reprogramming - It is possible. In certain cases, the compositions and methods provided herein are for fibrous disease Antifibrosis to fibrosis observed during the course of disease development (e.g., IPF and certain cancers) Reverse the shift towards promotion. In some embodiments, the compositions provided herein The method involves identifying fibrous biomarkers (e.g., fibrous biomarkers) in samples taken from individuals or subjects. Those related to fibrosis-promoting activity (e.g., CCL18, hydroxyproline, and Collagen) Reduces the amount / expression of the gene. When used herein, "individual" and "subject" also The "patient" is a mammal, preferably a human, but may also be an animal.

[0053] The terms “marker” or “biomarker” as used herein refer to active The level of expression in subjects experiencing a disease state is different from that in healthy subjects or subjects experiencing a disease state. If the expression level is significantly different from that of the subject or sample that is not being expressed, differential expression may occur. It can be described as follows. Differentially expressed markers may be normal or control samples, Or, compared to the baseline expression level of the subject, it is overexpressed or underexpressed (under (express) may be (in the embodiment described in the preceding paragraph, the biomarker is reduced) (Slightly or underexpressed). Increase or decrease in markers in biological samples, or quantification. This technique is used to measure the presence and / or relative abundance of gene products or transcripts. It can be determined by one of several methods known in the field. The marker level is absolute. It may be determined as a value or by comparing it to a baseline value, and also as a cutoff index (c It may be determined as the level of the target marker compared to the utoff index. Alternatively, the relative abundance of one or more markers may be clinically normal for the subject. It may be determined by comparison with a control. Furthermore, as used herein, the term "genetic hyper "Excessive expression" and "overexpression" (when used in relation to genes) and their formations This can lead to mutant phenotypes and / or result in abundant target protein expression. This includes, but is not limited to, the overexpression or misexpression of the wild-type gene product obtained, related It has meaning derived from the work of a person skilled in the field.

[0054] In some embodiments, the compositions and methods provided herein are anti-fibrotic bio Increasing marker levels (e.g., TNFα and IFN-γ). In some embodiments, , to reverse the M2-like phenotypic shift (for example, for fibrous diseases, disorders, or conditions) A composition is provided that provides an effective treatment.

[0055] In at least one embodiment, a drug comprising an immunomodulator is used as described herein. Compounds used in this method are produced. When used herein, "immunomodulatory compounds" "Quality" is achieved by inducing the activation or increased activity of one or more components of the immune system. Any drug, warhead that stimulates or otherwise affects the target immune system ( warhead), or other compositions or compounds. For example, not limited to Although not directly related, immunomodulatory substances target signaling pathways in immune cells. Compounds that target one or more pattern recognition receptors, in addition to or instead of the above. Alternatively, it may include a composition.

[0056] Examples of immunomodulatory substances in this disclosure include, but are not limited to, Toll-like receptors (TL receptors). R) agonist, interferon gene stimulant (STING), nucleotide ligation Syn-oligomeric domain (NOD)-like receptor (NLR), retinoic acid-inducible gene I ( RIG-I-like receptor (RLR), absent in melanoma elanoma 2 (AIM2)-like receptor (ALR), advanced glycation end products ( The receptor (RAGE) of the glycation end product, or the cell This disclosure includes any other pattern recognition receptors located within endosomes or cytoplasm. These immunomodulators, either additionally or alternatively, function further downstream in the pathway, in activated B cells. Inhibition of nuclear factor kappa-light chain enhancer (NFκβ) activator or Iκβ kinase It may contain agents. Table 1 shows such NFs that may be used as immunomodulators in this disclosure. Examples of κβ activators or Iκβ kinase inhibitors are provided. [Table 1] JPEG2026090286000020.jpg141170

[0057] As used herein, "TLR" refers to a protein that plays a role in the innate immune system. It is a type of pattern recognition receptor and is an example of a pattern recognition receptor. TLRs are structurally preserved receptors derived from microorganisms. It may be a single transmembrane receptor that recognizes the present molecule. TLRs are, for example, found in dendritic cells. macrophages, natural killer cells, adaptive immune cells (e.g., T lymphocytes and B lymphocytes). Leukemia includes lymphocytes and non-immune cells (epithelial cells, endothelial cells, and fibroblasts). They can be expressed on the sphere membrane. Non-limiting examples of TLRs include TLR1, TLR2, and TLR 3, TLR4, TLR5, TLR6, TLR7, TLR8, TLR9, TLR10, TL Examples include R11, TLR12, and TLR13. In some embodiments, this specification The TLR agonist provided in the book is coupled to one or more TLRs. Several embodiments Therefore, the TLR agonists provided herein are TLR7, TLR8, or TLR9 It binds to. In some embodiments, the TLR agonists provided herein are TL It binds to R7. In some embodiments, the TLR agonists provided herein are It binds to TLR7 and TLR8. In some embodiments, the agonist binds to the receptors. It is a ligand that binds to and activates the receptor.

[0058] Suitable for reprogramming activated macrophages (M2-like phenotype) to M1-like phenotype. Any therapeutic agent (e.g., a drug) may be used, and the drug (or warhead) may be used to treat the cells It may operate in the endosome and / or cytoplasm (for example, depending on its structure). In at least one embodiment, the therapeutic agent is an immunomodulator (e.g., pattern recognition receptors and that positively regulates the RIG-I / MDA5 pathway and / or its downstream signaling pathway (in each case, part of the innate immune system , for example, TLR, NLR, RLR, ALR, RAGE, and / or STING agonists, and / or Pelle / interleukin-1 receptor-associated kinase (IRAK) family, such as IRAK-M inhibitors, etc.). In other embodiments , the compounds provided herein include phosphoinositide 3-kinase (PI3K) kinase inhibitors, or other inhibitors that negatively regulate the adaptive immune system (e.g., can be used alone or in combination with immunomodulators that target pattern recognition receptors). In some embodiments , particularly when used for the treatment of IPF or other fibrotic conditions, the composition or compound (e.g., drug) includes (a) an immunomodulator that targets a pattern recognition receptor and / or an immunomodulator that is an agonist of the downstream signaling pathway of its innate immune system, and (b) a combination of mammalian rapamycin target protein (mTOR) inhibitors (ATP-competitive or otherwise), such as rapamycin or CZ415, etc. . In certain embodiments, as described herein, compounds are provided that include a targeting moiety (or a radical thereof) attached to an immunomodulator (or a radical thereof) that targets a pattern recognition receptor of a cell , and the targeting moiety includes a folic acid ligand or a functional fragment or analog thereof . "Folic acid" includes folic acid and analogs and derivatives of folic acid, such as folic acid, pteroylpolyglutamic acid, pteroyl-D-glutamic acid, and folic acid receptor-binding pteridines , for example, tetrahydropterin, dihydrofolic acid, tetrahydrofolic acid, and their deaza and dideaza analogs, but is not limited thereto .

[0059] . In particular embodiments, as described herein, compounds are provided that include a targeting moiety (or a radical thereof) attached to an immunomodulator (or a radical thereof) that targets a pattern recognition receptor of a cell , and the targeting moiety includes a folic acid ligand or a functional fragment or analog thereof . "Folic acid" includes folic acid and analogs and derivatives of folic acid, such as folic acid, pteroylpolyglutamic acid, pteroyl-D-glutamic acid, and folic acid receptor-binding pteridines , for example, tetrahydropterin, dihydrofolic acid, tetrahydrofolic acid, and their deaza and dideaza analogs, but is not limited thereto . .

[0060] The terms "deaza" and "dideaza" analogues refer to the presence of one or two nitrogen atoms in the natural folic acid structure. Analogues recognized in the art, or analogues thereof, having carbon atoms instead of elementary atoms. Alternatively, it refers to derivatives. For example, deaza analogs include folic acid, folic acid, and pteropolyglutamic acid. Minic acid, and leaf compounds such as tetrahydropterin, dihydrofolate, and tetrahydrofolate. Acid receptor-binding pteridines 1-deaza, 3-deaza, 5-deaza, 8-deaza, and 1 It may contain 0-dideaza analogs. Examples of dideza analogs include 1,5-didea of ​​folic acid. Examples include the 5,10-dideaza, 8,10-dideaza, and 5,8-dideaza analogs. In the context of this disclosure, other folic acids useful as complex-forming ligands are folic acid receptor ligands. Synthetic compounds: pemetrexed, proguanil, pyrimethamine, trimethoprim, pralatrex Sart, Lartitrexed, Aminopterin, Ametopterin (as methotrexate) (Also known as), N 10 -Methylfolate, 2-deaminodihydroxyfolate, deaza analog, e.g. For example, 1-deazametopterin or 3-deazametopterin, and 3',5'-diclo Ro-4-amino-4-deoxy-N 10 -Methylpteroylglutamic acid (dichlorometh Examples include trexate.

[0061] Folic acid, as well as the aforementioned analogs and / or derivatives, is also folic acid To reflect its ability to bind to receptors, "folate," "the folate in question," and These are called "multiple folates." As described herein, When the molecule is conjugated with an exogenous molecule, it induces folate-mediated endocytosis. It is effective in enhancing transmembrane transport, for example, through [a specific mechanism]. The aforementioned literature is described herein. It can be used in folate receptor-binding ligands. The term "ligand" refers to a component attached to the central atom or ion of a compound (e.g., a drug). It is a child, ion, or atom.

[0062] Specific embodiments of the novel compounds of this disclosure are provided herein. Those skilled in the art will understand this disclosure. It will be understood that compounds can exhibit polymorphism. In fact, the compounds of this disclosure possess useful properties. Any racemic, optically active, polymorphic, or otherwise exhibiting the characteristics of any of the compounds described herein. This may include stereoisomers or mixtures thereof, and a method for preparing optically active products (e.g., recombination Chiral synthesis by splitting racemic mixtures using crystallography, and by synthesis from photoactive starting materials. By means of, or by chromatography separation using a chiral stationary phase, and Using the standard tests described in the specification, or using other similar tests well known in the art Methods for determining antitumor activity using this method are well known in the art. Furthermore, a different method is available. Unless otherwise specified, the structures described herein also include all stereochemical forms of the structures. This means that each asymmetrical center configuration includes both right-hand (R) and left-hand (S) configurations. Therefore, the single stereochemical isomer and enantiomer and dias of this composition The Tromea mixture is within the scope of this disclosure.

[0063] The specific values ​​listed herein for radicals, substituents, and ranges are not specified otherwise. Unless otherwise specified, examples are provided for illustrative purposes only, and such examples illustrate radical and substitution. For the radical, other defined values or other values within the defined range are not excluded. For example , (C1-C6)alkyl can be methyl, ethyl, propyl, isopropyl, butyl, iso -butyl, sec-butyl, pentyl, 3-pentyl, or hexyl, (C1-C3)alkyl can be iodomethyl, bromomethyl, chloromethyl, fluoro methyl, trifluoromethyl, 2-chloroethyl, 2-fluoroethyl, 2,2,2-tri fluoroethyl, or pentafluoroethyl, (C1-C3)alkoxy can be methoxy, ethoxy, or propoxy, (C2-C6 )alkanoyloxy can be acetoxy, propanoyloxy, butanoyloxy, isob utanoyloxy, pentanoyloxy, or hexanoyloxy.

[0064] Furthermore, when a moiety is substituted with an R substituent or a substituent, the group may be referred to as "R-substituted". When a moiety is described as being either R-substituted or otherwise generally containing a substituent , the moiety is substituted with at least one R substituent, and each substituent is optionally different. The substituent (or R substituent) may contain any molecule or combination of molecules but its inclusion does not substantially affect the overall structure and shape of the compound and does not change any hydrogen bonds essential to the compound for achieving its intended purpose (e.g., binding to a target pattern recognition receptor).

[0065] It will be understood that any molecule or combination of molecules may be included as long as they do not change any hydrogen bonds essential to the compound for achieving its intended purpose (e.g., binding to a target pattern recognition receptor).

[0065] When substituents are specified by the conventional chemical formula written from left to right, they are, for example For example, -CH2O- is equivalent to -OCH2-, and the structure is described from right to left. It similarly includes the chemically identical substituents that result.

[0066] In some embodiments, the immunomodulators of the compounds provided herein (e.g., TL) The R7 agonist group has the structure of formula XX, more specifically, the structure of formula XX'. It is; [ka] During the ceremony, R 1B -NH2 or -NH-R 1X And, R 2B These are hydrogen (H), alkyl, alkenyl, alkynyl, alicyclic, aryl, and vial. rheu, heteroaryl, -NH-R 2X , -OR 2X , -SR 2X , [ka] And, R 1X , R 2X , and R 2Y Each of these can independently be H, alkyl, alkenyl, or alkini. Selected from the group consisting of aryl, alicyclic, aryl, biaryl, and heteroaryl compounds, [ka] These are 3-10 member nitrogen-containing non-aromatic monocyclic or bicyclic heterocyclic rings. X is either CH or nitrogen (N).

[0067] As used herein, alkyl, alkoxy, etc. are linear (i.e., unbranched) Alternatively, it may indicate branched chains, or combinations thereof, which are fully saturated, monovalent, or It may be polyunsaturated, and may contain divalent and polyvalent radicals, and may be a specified carbon source It has a number of children (i.e., C1-C 10 (This refers to 1 to 10 carbon atoms). Saturated carbonization Examples of hydrogen radicals are not limited to methyl, ethyl, and n- Propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl, (C) Chlohexyl(methyl), e.g., n-pentyl, n-hexyl, n-heptyl, n-oc Examples include homologous and isomeric groups such as til. Unsaturated alkyl groups include one or more double groups. It is an alkyl group having a bond or triple bond. An example of an unsaturated alkyl group is vinyl , 2-propenyl, clotyl-2-isopentenyl, 2-(butadienyl), 2,4-pe Ntadienyl, 3-(1,4-penadienyl), ethinyl, 1- and 3-propynyl, This includes, but is not limited to, 3-butynyl, as well as higher homologs and isomers. No. The alkoxy is attached to the rest of the molecule via an oxygen linker (-O-). It is alkoxy. In some embodiments, the oxygen atom of formula -O-alkyl is This refers to radicals that are bonded via this medium.

[0068] Generally, the term "acyl" or "acyl substituent" refers to one or more substituents from an oxoacid, including inorganic acids. This refers to the derivative obtained by removing the hydroxyl group above, and the double bond oxygen atom It contains alkyl groups. Furthermore, references to individual radicals such as "propyl" are direct It includes only chain radicals, and specifically refers to branched-chain isomers such as "isopropyl" that are referenced. It is mentioned.

[0069] In some embodiments, the TLR7 agonist has formula X, and the TLR7 agonist is R via linker 1A , R 1B , R 3A , or R 3B In any one of the following If the targeting portion is conjugated and the TLR7 agonist has formula XX', then TLR 7. The agonist is linked through the linker R 1A , R 1B , R 3A , or R 3B one of them It is then conjugated to the targeted portion.

[0070] As used herein, the term “linker” means a chemical bond formed with A, B, or S. It is biofunctionally adapted to achieve the function of connecting two or more functional parts of a molecule to achieve the function of this disclosure. It contains a chain of atoms that forms a compound. For example, the chain of atoms is carbon (C), N, oxygen (O) ), sulfur (S), silicon (Si), and phosphorus (P), or C, N, O, S, and P The atoms can be selected from C, N, O, and S. The chain of atoms is used in compounds such as folic acid and pharmaceuticals. Different functional capabilities can be covalently bonded. A linker consists of approximately 2 to 100 atoms in a continuous skeleton. It may include a wide variety of links, such as the range of links, and may include releaseable or non-releaseable linkers. In some embodiments, immunomodulators of compounds provided herein (e.g., The TLR7 agonist group has the structure of formula XXX, more specifically, the structure of formula XXX'. It is a radical that does, [ka] During the ceremony, R 1C-NH2 or -NH-R 1X And, R 2C This is combined, NH, -NR 1X , or CH2 And, If applicable, [ka] These are 3-10 member nitrogen-containing non-aromatic monocyclic or bicyclic heterocyclic rings. X A This is CH2, NH2, or -NH-R 1X And, Each R 1X These are independently H, alkyl, alkenyl, alkynyl, alicyclic, aryl, and via. Selected from the group consisting of reels and heteroaryls, The TLR7 agonist is R via the linker. 1C , R 2C , or R 3B one of them It is conjugated to the targeted portion.

[0071] In some embodiments, the compound has a linker between the targeting moiety and the immunomodulator, Otherwise, connect the linker ("L" or "L" n This further includes "). How many In that embodiment, linker L n It is configured to avoid the release of immunomodulatory substances, n is an integer less than or equal to 50. In some embodiments, the linker L n is polyethylene glycerin Includes a PEG linker or a PEG derivative linker, where n is in the range of 1 to 32. The selected integer is specific to the folate receptor β. Several implementations Morphologically, n can be 1-50, 1-10, 2-8, or 2-4.

[0072] In some embodiments, L is a hydrolyzable linker. In some embodiments, L is a hydrolysis-inhibiting linker. It is a substituted heteroalkyl group.

[0073] The term "alkylene" is used alone or as part of another substituent, unless otherwise specified. While exemplified by -CH2CH2CH2CH2-, it is derived from alkyl groups and is not limited to this. This refers to a divalent radical. Typically, an alkyl (or alkylene) group has 1-2 It has four carbon atoms. "Lower alkyl" or "lower alkylene" refers to short-chain alkyl Alternatively, it is an alkylene group, generally having eight or fewer carbon atoms.

[0074] The term "heteroalkyl" is used alone or in combination with other terms unless otherwise specified. As long as at least one carbon atom and selected from the group consisting of O, N, P, Si, and S A stable linear or branched chain composed of at least one selected heteroatom, This means the combination, where the nitrogen and sulfur atoms may optionally be oxidized, and the nitrogen hetero Atoms may be optionally quartered. Heteroatoms O, N P, S, and Si are at any internal position of the heteroalkyl group, or the alkyl group is molecular It may be positioned to adhere to the remaining part. Examples include, but are not limited to, However, -CH2-CH2-O-CH3, -CH2-CH2-NH-CH3,-CH2-C H2-N(CH3)-CH3,-CH2-S-CH2-CH3,-CH2-CH2-S( O)-CH3,-CH2-CH2-S(O)2-CH3,-CH2=CH-O-CH3, -Si(CH3)3,-CH2-CH=N-OCH3,-CH=CH-N(CH3)-C Examples include H3, -O-CH3, -O-CH2-CH3, and -CN. Up to two heliums. The terror atoms may be consecutive, for example, in a structure like -CH2-NH-OCH3.

[0075] Similarly, the term "heteroalkylene" refers to itself or as part of another substituent. Unless otherwise specified, for example, -CH2-CH2-S-CH2-CH2 and -CH2 Heterozygous hetero This refers to a divalent radical derived from an alkyl group. For heteroalkylene groups, the heteroprimary radical is used. The child also has chain ends (e.g., alkylene oxy, alkylenedioxy, alkylene amino It may occupy either or both of the following (such as alkylenediamino acids). Furthermore, it may also contain For chilen and heteroachilen linking groups, depending on the direction in which the linking group formula is written, The orientation of the linking group is not implied. For example, the formula -C(O)2R'- is -C(O)2R'- It represents both -R'C(O)2' and -R'C(O)2'. As mentioned above, heteroalkyl groups are used herein. When used, for example, -C(O)R', -C(O)NR', ​​-NR'R'', -OR', -SR' and / or -SO2R' are attached to the rest of the molecule via heteroatoms. Includes the groups to be attached. "Heteroalkyl" is listed, followed by specific groups such as -NR'R''. When a list of heteroalkyl groups follows, the terms heteroalkyl and -NR'R" are used. It will be understood that they are not redundant or mutually exclusive. Rather, to add clarity, A list of specific heteroalkyl groups is provided. Therefore, the term "heteroalkyl" is used herein. So, should this be interpreted as excluding certain heteroalkyl groups such as "-NR'R"? do not have.

[0076] In some embodiments, L is alkyl, hydroxyl, oxo, PEG, or carboxy Substitutive heterotypes containing at least one substituent selected from the group consisting of silates and halos It is a rhalkyl compound. "Halo" or "halogen" is either by itself or as part of another substituent. Unless otherwise specified, this refers to fluorine, chlorine, bromine, or iodine atoms.

[0077] In some embodiments, L is a spacer (for example, as described elsewhere in this specification). (to include) In some embodiments, the spacer is peptidoglycan or sugar. Includes.

[0078] In some embodiments, L has at least one disulfide bond in its skeleton. It is a substituted heteroalkyl. In some embodiments, L is present in its skeleton at least It is also a peptide that has one disulfide bond.

[0079] The terms “polypeptide,” “peptide,” and “protein” are used interchangeably in this specification. Substitutably used, polymers, polypeptides, or polypeptides of amino acid residues, peptides This term refers to a fragment of a fusion polypeptide, or a fusion polypeptide. This term refers to a molecule consisting of one or more amino acid residues. Amino acid polymers, which are artificial chemical mimics of corresponding naturally derived amino acids, and This applies to naturally derived amino acid polymers and non-naturally derived amino acid polymers.

[0080] In some embodiments, L is -CONH-CH(COOH)-CH2-SSC H2-CRa R b -O-CO-, -CONH-CH(COOH)CR a R b -O-CO- ,C(O)NHCH(COOH)(CH2)2-CONH-CH(COOH)CR a R b -O-CO- or -C(O)NHCH(COOH)(CH2)2-CONH-CH(C OOH)-CH2-SS-CH2-CR a R b -O-CO- is included in the formula, where R a and R b These are independently H, alkyl, or heteroalkyl (e.g., PEG).

[0081] In some embodiments, L includes the following structure: [ka] In the formula, n and m are independently between 0 and 10.

[0082] In some embodiments, L includes the following structure: [ka] In the formula, n is 1 to 32. In at least one exemplary embodiment, n is 1 to 30. Yes, w is between 0 and 5.

[0083] In some embodiments, L includes the following structure [ka] In the formula, n is between 1 and 30, and w is between 0 and 5.

[0084] In some embodiments, the compound has a structure represented by the following formula. [Chemical Formula]

[0085] In some embodiments, the compound has a structure represented by the following formula. [Chemical Formula]

[0086] In some embodiments, the compound has a structure represented by the following formula. [Chemical Formula]

[0087] In some embodiments, the compound has a structure represented by the following formula. [Chemical Formula]

[0088] In certain embodiments, provided herein is a compound comprising a folic acid ligand or a functional fragment or analog thereof attached via a linker to an immunomodulatory substance comprising a TLR agonist and having a targeting moiety, wherein the TLR agonist has the following structure represented by formula XXX and has the following structure [Chemical Formula] wherein, R 1 is an amine group, R 2 is a single bond -NH-, R 3 is H, alkyl, hydroxy group, or any other substituent thereof, and X is CH2, NH, O, or S and Yes, the linker is attached at R 1 , R 2 , or R 3 . Additionally or alternatively , R1 may be -NH2 or -NH-R 1X , and R2 may be H, alkyl, a lkene, alkyne, alicyclic, aryl, biaryl, heteroaryl, -NH-R2 X , -O-R 2X , -S-R 2X ,

Chemical formula

Chemical formula

[0089] In some embodiments, what is provided herein is a pharmaceutical composition comprising any of the formulas or compounds provided herein, the linker comprises PEG or a PEG derivative, and in some cases, is a non-cleavable linker attached at R 3 , or is a cleavable linker attached at R 1 , R 2 , or R 3 .

[0090] In some embodiments, pharmaceutically acceptable salts are hydrobromide, citrate, and triglycerides. Fluoroacetate, ascorbic acid, hydrochloride, tartrate, triflate, maleate, me Selected from silates, formates, acetates, or fumarates.

[0091] In some embodiments, the compound is a TLR agonist (e.g., its radical), e.g. For example, although not limited to, TLR3 agonist, TLR7 agonist, TLR7 / 8 agonist, TLR8 agonist, or TLR9 agonist (for example, all of them It includes (which binds to Toll-like receptors present in the cell's endosomes). For example, limited Although not definitively defined, in at least one exemplary embodiment, the immune response of the drug / compound The modifier may be selected from the compounds listed in Table 2 below. [Table 2] JPEG2026090286000037.jpg235170JPEG2026090286000038.jpg230170JPEG2026090286000039.jpg253170JPEG2026090286000040.jpg99170

[0092] In some embodiments, the compounds provided herein are compounds of formula I (or radi). (For example, a TLR7 agonist), [ka] or a pharmaceutically acceptable salt thereof or containing thereof. In some embodiments, R 1 teeth It is an amine. In some embodiments, R 2 is (for example, a single) bond, or amine (For example, -NH-). In a particular embodiment, R 3 H, alkyl, hydroxy A group, or any other suitable substituent (e.g., those described herein). In some embodiments, X is CH2, NH, O, or S. The compounds provided herein comprise a radical of formula I, and the targeting moiety is any appropriate At a certain position, for example, R 1 , R 2 , and / or R 3 And, or through these (for example, They are conjugated or linked together (via a linker and / or directly).

[0093] In at least one exemplary embodiment, the compounds described herein are compounds of formula Ia. (or radicals) (e.g., TLR7 agonists) [ka] or a pharmaceutically acceptable salt thereof, or comprising the same. In some embodiments, X is CH or N. In some embodiments, R1 is -NH2 or -NH- R 1X In some embodiments, R2 is H, alkyl, alkenyl, or alkini. aryl, biaryl, heteroaryl, -NH-R 2X , -OR 2X , -SR 2X , [ka] In a particular embodiment, R 1X , R 2X , and R 2Y Each of these is independent of H, A Lukyl, alkenyl, alkinyl, alicyclic, aryl, biaryl, and heteroaryl Selected from the group consisting of . In some embodiments, [ka] This is a 3- to 10-membered nitrogen-containing non-aromatic monocyclic or bicyclic heterocycle. Several embodiments The compounds provided herein comprise the radical of formula Ia, and the targeting moiety is any In the appropriate position, for example, R 1 , R 2 , and / or R 3 And, or through these (for example) If, then, via a linker and / or directly, they are conjugated or linked.

[0094] In some embodiments, the compounds provided herein are compounds of formula II (or L). (For example, a TLR7 agonist), [ka] or a pharmaceutically acceptable salt thereof, or comprising the same. In some embodiments, , R 1 is an amine. In some embodiments, R 2 is (for example, a single) bond or It is -NH-. In certain embodiments, R 3 is H, alkyl, hydroxyl group, or any Other suitable substituents, for example, are described herein. Some embodiments Then, X is CH2, NH, O, or S. In some embodiments, as specified herein The provided compound contains the radical of formula II, and the targeting moiety is at any suitable position, for example. Eba R 1 , R 2, and / or R 3 And, or via these (for example, via a linker) To be conjugated or combined (and / or directly).

[0095] In other embodiments, the compounds disclosed herein are TLR agonists of formula III (e.g., or The agent may include the radical (or a pharmaceutically acceptable salt thereof), [ka] In the formula, R 1 is an amine group, R 3 This is a hydroxyl group. Furthermore, if desired, The targeting portion (e.g., or its radical) or other ligands are R 1 or R 3 smell (either via the linker or directly) it is conjugated to the agonist of Equation III. It is also possible. The TLR agonist of formula III (for example, or its radical) is TLR7 It is a gonist and is at least 10 times (10x) more powerful than conventionally available TLR7 agonists. That is the case.

[0096] In some embodiments of this specification, a TLR7 agonist of formula IV (for example, or so A radical of (or a pharmaceutically acceptable salt thereof) or a pharmaceutically acceptable salt thereof is provided. [ka] In the formula, R 1 is an amine group, R 2 It is a single bond -NH-.

[0097] In some embodiments, the administration of the compounds provided herein (e.g., to an individual) This converts macrophages in fibrous tissue from an M2-like phenotype to an M1-like phenotype. In the embodiment, after administration of the compounds provided herein, collagen synthesis is stimulated. A decrease in tokines (i.e., CCL18, PDGF, and IL-1β) occurs, Simultaneously, an increase in cytokines that inhibit collagen production (e.g., IFN-γ) occurs. In particular, in at least one embodiment, after administration of the compound provided herein, The itokine profile matches the reprogramming of the M2-like phenotype to the M1-like phenotype. As used herein, "profile" or "assay" refers to one or more marks. A set of cars, and their presence, absence, and / or (for one or more contrasts) This refers to the relative level or abundance of substances present in the sample. For example, a cytokine profile is a relative level or abundance of substances present in the sample. This is a dataset of the presence, absence, relative levels, or abundance of cytokines. The nucleotide or nucleic acid profile indicates the presence or absence of expressed nucleic acids (e.g., transcripts, mRNA, etc.). This is a dataset of present, relative, or abundant levels. Alternatively, a profile is an expression profile. It is sometimes called a rofile.

[0098] In some embodiments, the net result of reprogramming is the alveolar sacs. Increased r air sac), decreased extracellular matrix deposition, and hydroxyproline / collagen This results in a decrease in enzyme biosynthesis and effective recovery from the disease (see, for example, Example 4).

[0099] While specific drugs and formulations are described herein, activated myelocytes are anti-fibrotic M1 Any compound (e.g., a drug) that is useful for reprogramming to a specific phenotype is described herein. Novel compounds and methods (e.g., binding to pattern recognition receptors and their downstream innate immune system) Any compound (e.g., a drug) that can inhibit at least part of the response can be used. It should be understood that in some embodiments, the compounds described herein may be used. Analogues and / or derivatives thereof may be used in the targeted compounds provided herein.

[0100] Furthermore, two or more compounds can be administered, and in some cases, the compounds may be different drugs. This may include different drugs such as TLR7 agonists and TLR9 agonists. It can be selected from nist. In yet another embodiment, one or more compounds are one The above conjugated and / or unconjugated drugs (e.g.) It can be administered in the composition together with the conjugated embodiments described below. In some embodiments, any of the compounds and agents described herein are used as described herein. It can be used according to the method described, and in some cases, depending on the desired use, bone Depletes or inhibits myelin-derived suppressor cells (e.g., in connection with cancer treatment) and growth factors Downregulating the production of (for example, pirfenidone, which is associated with the treatment of IPF) , mammalian rapamycin target protein complex 1 (mTORC1) signaling (for example) By inhibiting CZ415, which is associated with the treatment of IPF or other fibrotic conditions, the fibrosis Other drugs that directly modify blast cells, and / or any other antifibrotic and / or It can be used in combination with anticancer drugs and therapies. When used herein, "down Regulation" and its constituent elements (e.g., "down-regulation" or " "Down-regulated" (etc.) may be used interchangeably, and genes, nucleic acids This refers to a decrease in the levels of markers such as metabolites, transcripts, proteins, or polypeptides. When used herein, "upregulation" and its formations (for example) (e.g., "p-regulation" or "up-regulated") is compatible. It may be used for purposes such as genes, nucleic acids, metabolites, transcripts, proteins, or polypeptides. This refers to an increase in the levels of markers such as d, as well as pathways such as signal transduction or metabolic pathways. They can also be up- or down-regulated.

[0101] targeting part In some cases, this relates to the systemic administration of at least one conventional drug as specified herein. Toxicity is practical in relation to the treatment of fibrotic diseases, cancer, or any other disease state. This hindered application. For example, TLR agonists may not be resistant to certain individuals, and in some cases, In some cases, it may result in the death of the subject (for example, when administered systemically via conventional methods). (If...). In some embodiments, compounds provided herein, for example, of formula I and Compounds having / or II are more effective than conventional drugs that can be used in combination with the compounds of this disclosure. It is also extremely potent, and in some cases, a mechanism to avoid systemic toxicity is desirable.

[0102] In certain embodiments, the targeting portion and the conjugated therapeutic agent (for example, (as described above) Such drugs are provided herein. In some embodiments, the targeted portion ligands that target specific regions or tissues of an individual (e.g., those with high specificity) or include other atoms or molecules, in certain examples, for example, hormones, antibodies, and / or It may contain vitamins. As will be explained in more detail below, at least one implementation Morphologically, the targeting portion contains a molecule that has (e.g., high) affinity for FRβ. In some cases, the targeted area may be cells or tissue from fibrous disease or cancer, as needed. It possesses specific affinity for any particular receptor.

[0103] In some cases, FRβ is activated bone marrow cells (e.g., mainly activated monocytes). In M2-like macrophages, and in other cells, the regulation was significantly upregulated, compared to previous methods. All recorded data indicates that FRβ is myeloid after exposure to anti-inflammatory or pro-inflammatory stimuli. This supports the idea that it is induced only in the source cells. Folate receptors are involved in non-mucinous ovarian cancer. It can be upregulated to (for example, over 90%). In certain cases, folate intake The condition is present in the kidneys, brain, lungs, and breast cancer. For example, it itself provides the desired specificity. There are many cancers that do not express enough folate receptors to serve, but cancerous tumors, for example For example, express bone marrow-derived suppressor cells (MDSCs) that express FRβ, as described in this specification. It can be targeted by the targeting portion provided in the book. In some embodiments, Folate receptors are found in healthy (non-myelocytic) tissues (e.g., lungs, liver, spleen, heart, brain, muscles, intestines). It is virtually absent (for example, present only at very low levels) in organs such as the pancreas and bladder. In some cases, even quiescent macrophages, which are abundant throughout the body, are primarily FR. It is β-negative. In some cases, the uptake of folate-targeted contrast agents is, for example, in inflammatory tissue. Malignant lesions, and located in the kidneys. In certain cases, non-cancerous subjects have lesions in the kidneys and inflammatory sites. It retains only folate-targeted drugs. In some cases, mismatch in folate receptor expression can lead to fibrosis. It provides a mechanism for selectively targeting rhinocytes.

[0104] In some embodiments, the compounds and methods provided herein are limited to FRβ By utilizing this expression, potent compounds (e.g., conjugates or drugs) administered systemically can be used. To target / localize to fibrous and / or cancerous tissue. In some cases, as specified herein The provided compounds, for example, favorably prevent systemic activation of the immune system in FRβ-expressing cells. Direct delivery and, for example, systemic use of non-target compounds (e.g., drugs) described herein. This allows us to avoid the toxicity (for example, at least some of it) that has previously hindered this process. In some embodiments, the methods described herein may, for example, determine whether cancer expresses folate receptors. Regardless of whether it is used to treat fibrous diseases and / or cancer, how many In one embodiment, folic acid and other folic acid receptor-binding ligands such as folic acid (or so The radical (for example) has affinity for FRβ and is therefore used as a targeting moiety. It can be done.

[0105] Folic acid is a member of the vitamin B family and is used, for example, in the production of nucleic acids and amino acids. By participating in synthesis, folic acid can play a crucial role in cell survival. This refers to the specificity of immunomodulatory drugs conjugated by targeting activated bone marrow cells. By enhancing and targeting folate receptor-positive cancer cells, the conjugated anti- The drug can be enhanced. In some cases, the targeted portion and immunomodulatory substances (e.g.) (as a TLR7, TLR8, TLR7 / 8, TLR9, or TLR3 agonist), Compounds comprising folic acid ligands (or their radicals), or functional fragments or analogs thereof. The materials are provided in this specification. In some cases, TLR7, TLR8, TLR7 / 8, TL R9 and TLR3 are present in the endosome. In some embodiments, the compound, Or the radical binds to the TLR. In some implementations, the TLR binds to the TLR. The answer is 7.

[0106] Instead, pyrido[2,3-d]pyrimidine analog ligands (e.g., or its radiols) Cal), its functional fragment or analogue, or affinity of FRβ (e.g., not limited to) However, any other molecule, fragment, or atom having high specificity can be targeted (or otherwise targeted). It may also be used as a radical of . For example, such a folic acid analog molecule can be used at approximately 20°C / At 25°C / 30°C / physiological temperatures, it binds approximately 0.01 or more FRβ compared to folic acid. It may have relative affinity for Galectin-3 ligand, translocate TSPO ligands, and fibrous and / or cancerous cells or groups Any other ligand or targeting moiety having a highly specific affinity for the tissue is employed. It's okay.

[0107] Specific examples of suitable targeting moieties (or radicals) are provided herein, but this disclosure The targeting portion (or its radical) is any ligand useful for targeting FRβ. (Or its radicals) may be included, and is not limited to the structures specified herein. This should be understood. The ligand (or its radical) can bind to FRβ.

[0108] In at least one embodiment, the compounds provided herein have a structure of formula V or so A targeting moiety (or its radical) having a functional fragment or analogue thereof, [ka] During the ceremony, X1, X2, X3, X4, X5, X6, X7, X8, and X9 are each independent of N , NH, CH, CH2, O, or S, Y is C, CH, CH2, N, NH, O, or S. Z is glutamate, valine, or a substrate. R1 and R2 are independently NH2, OH, SH, CH3, or H. R3 is time or alkyl, m and n are independently 0, 1, or between 0 and 1. [ka] This represents either a single bond CC or a double bond CC.

[0109] In a further embodiment, as a non-limiting example, the targeting portion (or radical) of formula V is , having the structure of VI (or its functional fragment or analogue), [ka] During the ceremony, X1, X2, X3, X5, X6, X7, X8, and X9 are each independently N, NH, It is CH, CH2, O, or S, Y is C, CH, CH2, N, NH, O, or S. Z is glutamate, valine, or a substrate. R1 and R2 are independently NH2, OH, SH, CH3, or H. R3 is time or alkyl, m and n are independently 0, 1, or between 0 and 1. [ka] This represents either a single bond CC or a double bond CC.

[0110] Another specific targeting portion of formula V (or its functional fragment or analogue) The radical has the structure of formula VII, [ka] During the ceremony, X1, X2, X3, X4, X5, X6, X7, X8, and X9 are each independent of N , NH, CH, CH2, O, or S, Y is C, CH, CH2, N, NH, O, or S. Z is glutamate, valine, or a substrate. R1 and R2 are independently NH2, OH, SH, CH3, or H. R3 is time or alkyl, m and n are independently 0, 1, or between 0 and 1. [ka] This represents either a single bond or a double bond (CC).

[0111] In some embodiments, the targeting portion of formula VI (or its radical) is formula VIII Having a structure, [ka] During the ceremony, X1, X2, X3, X5, X6, X7, X8, and X9 are each independently N, NH, It is CH, CH2, O, or S, Y is C, CH, CH2, N, NH, O, or S. Z is glutamate, valine, or a substrate. R1 and R2 are independently NH2, OH, SH, CH3, or H. R3 is time or alkyl, m is 0, 1, or between 0 and 1. [ka] This represents either a single bond or a double bond (CC).

[0112] In some embodiments, the targeting portion of formula VI (or its radical) is the structure of formula IX. It has a structure, [ka] During the ceremony, X1, X2, X3, X5, X6, X7, X8, and X9 are each independently N, NH, It is CH, CH2, O, or S, Y is C, CH, CH2, N, NH, O, or S. Z is glutamate, valine, or a substrate. R1 and R2 are independently NH2, OH, SH, CH3, or H. R3 is time or alkyl, m is 0, 1, or between 0 and 1. [ka] This represents either a single bond or a double bond (CC).

[0113] In some embodiments, the targeting portion of formula VII (or its radical) is formula X or It has the structure of XI, [ka] During the ceremony, X1, X2, X3, X4, X5, X6, X7, X8, and X9 are each independent of N , NH, CH, CH2, O, or S, Y is C, CH, CH2, N, NH, O, or S. Z is glutamate, valine, or a substrate. R1 and R2 are independently NH2, OH, SH, CH3, or H. R3 is time or alkyl, m is 0, 1, or between 0 and 1. [ka] This represents either a single bond or a double bond CC, or [ka] During the ceremony, X1, X2, X3, X4, X5, X6, X7, X8, and X9 are each independent of N , NH, CH, CH2, O, or S, Y is C, CH, CH2, N, NH, O, or S. Z is glutamate, valine, or a substrate. R1 and R2 are independently NH2, OH, SH, CH3, or H. R3 is time or alkyl, m is 0, 1, or between 0 and 1. [ka] This represents either a single bond or a double bond (CC).

[0114] Some additional embodiments of the targeted portion (e.g., or its radical) of the present disclosure The structural and spectral data are provided in Tables 3, 4, 5, and 6 below.

[0115] Table 3 shows the addition of a targeting moiety (e.g., or its radical) having the structure of formula VIII. This provides non-limiting examples of the embodiment. [Table 3] JPEG2026090286000063.jpg254170JPEG2026090286000064.jpg221170

[0116] Table 4 shows additional targeting moieties (e.g., or radicals) having the structure of formula IX. This provides non-limiting examples of embodiments. [Table 4] JPEG2026090286000066.jpg106170

[0117] Table 5 provides non-limiting examples of additional embodiments of the targeting portion having the structure of formula X. . [Table 5]

[0118] As mentioned above, instead of folic acid, the targeting part (for example, its radical) is, for example, pi Lido[2,3-d]pyrimidine or any of the formulas listed in Table 6 below (for example, the radical of the formula) One or more non-classical folate antagonists, such as similar analogs (or their radicals) that have ) It may also be an antidote analog (or an analog or functional fragment thereof). [Table 6] JPEG2026090286000069.jpg219170JPEG2026090286000070.jpg219170JPEG2026090286000071.jpg254170JPEG2026090286000072.jpg87170

[0119] In some cases, the compounds provided herein may have a targeted moiety (e.g., its radical) ) and conjugated drugs (e.g., their radicals) (e.g., immunomodulators) Hmm. Immunomodulators (e.g., their radicals) target the radical (e.g., their radical) and Directly, or via a linker (e.g., optionally including spacers) It may be gated. Figure 1A shows at least one embodiment of compound 100. Here Compound 100 is, for example, an immunomodulator having formula I (or its drug or radica (L)102 is included, in the formula, R 3 It is a hydroxyl group. Immunomodulatory substances (for example, its radio The CAL 102 is transmitted to the target portion (e.g., its radical) 104 via the linker 106. It is conjugated. Here, the targeted portion (e.g., its radical) 106 is conjugated with folic acid. Yes, (for example, non-release) linker 106 is a PEG linker repeated n times. n is between 1 and 32.

[0120] In at least one embodiment, and not limited to, compound 100 is It can be expressed by the following formula, QLT, where Q is the folate receptor binding ligand / targeting ligand. Part 104 is a radical, L is linker 106, and T is a TLR agonist / It is a radical of immunomodulator 102. Linker L is a linker as presented herein. It may include any of the formulas.

[0121] Similarly, Figure 1B shows at least one embodiment of compound 150. Compound 150 is For example, via the (e.g., releaseable) linker 156, the targeted portion (e.g., its A TLR7 agonist having formula III, for example, conjugated to (Dical) 154 (For example, the radical) of an immunomodulatory substance / drug (for example, the radical) 152 To possess.

[0122] Linker (L or L n ) may be releaseable or not releaseable. In some cases, the target of a compound containing a non-release linker may be, for example, an endosome (for example) (For example, the target cells) and on the other hand, the target of the releaseable linker is, in some cases, This could be endosomes, cytoplasm, or both (for example, of the cell in question).

[0123] In at least one exemplary embodiment, linker L n is the targeted part (for example, that A radical is positioned between an immunomodulator or a pharmaceutically acceptable salt thereof, and phosphorus Car L or L n It is configured to avoid the release of the free form of the TLR7 agonist. n is an integer less than or equal to 50. Additionally or alternatively, the compound may be PEG or a PEG derivative. Linker L containing the conductor n It may include n, and n is an integer selected from the range of 1 to 32. Even if that's the case, the targeting part (for example, its radical) is a folate receptor containing an FRβ-binding ligand. It may contain a radical of a solution-binding ligand.

[0124] The term "releaseable" in relation to linkers is used, for example, in relation to reducing agent instability and pH instability. Sex, acid instability, base instability, oxidative instability, metabolic instability, biochemical instability, enzyme Instability, or p-aminobenzyl-based polyvalent releaseable bond (p-aminoben zylic based multivalent releasable bond) It can be broken down under physiological conditions such as (e.g., by chemical or enzymatic hydrolysis) (ru) Means a linker containing at least one bond. A physiological condition that leads to the breakdown of a bond. The matter does not necessarily involve biological or metabolic processes; instead, for example, Organelles such as endosomes that have a physiological pH or a pH lower than cytoplasmic pH. It is understood that this may include standard chemical reactions such as hydrolysis reactions resulting from compartmentalization. The severable linkage is, for example, at either or both ends of the releaseable linker, as specified herein. As described, connect two adjacent atoms in the releaseable linker, and / or It can connect to other linker parts or targeting parts and / or drugs. In some cases, the releaseable linker is split into two or more fragments. The output linker is separated from the targeting portion. In some embodiments, the targeting portion These immunomodulatory substances are released from each other, and the immunomodulatory substances are activated.

[0125] In contrast, the term "non-release" associated with linkers is easily used under physiological conditions. This means a linker that contains at least one link that does not break quickly. Morphologically, an unreleased linker contains a skeleton that is stable under physiological conditions (e.g., skeleton). It is less susceptible to hydrolysis (e.g., aqueous hydrolysis or enzymatic hydrolysis). In one embodiment, the composition provided herein, which includes a non-release linker, is a composition of either component (for example, a targeted ligand (for example, a completely amorphous (FA)-ligand) or does not release immunomodulatory substances (e.g., TLR7 agonists). Several embodiments So, the non-release linker is a disulfide bond (e.g., SS) or an ester in the skeleton. Lacking a component. In some embodiments, the composition is used throughout the entire cycle of the composition (for example) (For example, during endocytosis to target cell endosomes), the scaffold remains virtually stable. Therefore, it includes a targeting portion and an immunomodulator that are connected. Compositions containing non-linking agents include immunomodulators that are present in TLRs within the endosomes of cells. It is particularly beneficial when targeting NOD-like receptors and / or other pattern recognition receptors. Yes, non-release linkers include amides, esters, ethers, amines, and / or thios. This may include ethers (e.g., thio-maleimide). Specific examples are provided herein. However, under physiological conditions, at least one bond is formed that does not break easily or rapidly. It will be understood that, provided that this condition is met, any molecule can be used with a non-release linker.

[0126] Perhaps more specifically, the non-release linker is, for example, at a neutral pH, for example, 10% -less than 10% (e.g., less than 5%, less than 4%, less than 3%, less than 2%, less than 1%) , less than 0.1%, less than 0.01%, or 0.001%) in an aqueous solution (e.g., buffered Hydrolyzing in a solution (for example, phosphate buffer) within a certain period (for example, 24 hours) Includes a linker. In some embodiments, when a non-dischargeable linker is used, (for example) Conjugate compounds administered (in systemic circulation before uptake by target cells / tissues) Approximately less than 10 percent (10%), preferably less than 5 percent (5%), or There is none, and the drug is released free. In some embodiments, within one hour of administration, the compound While the substance is in the systemic circulation, less than 5 percent (5%) of the free drug is released from the conjugate. It can be done.

[0127] In some embodiments, the targeting portion is such that the compound is therapeutically effective in vivo. Therefore, it is not cleaved from the drug / immunomodulator. In some embodiments, this is, for example, (Even if there are any) the drug (for example, an immunomodulator, for example, a TLR7 agonist) Because only a small amount is released before the compound's target delivery (e.g., systemically), it is potent. This is advantageous as it enables the use of targeted compositions containing drugs (e.g., TLR7 agonists). In some embodiments, adjusting the release characteristics of the active ingredient is effective for developing a pharmaceutical product. This describes a difficult aspect of preparing the product. In some embodiments, the emission-free products provided herein Compositions containing a linker avoid difficulties in preparing effective pharmaceutical compositions (e.g., release (By eliminating the need for timing). In some embodiments, as specified herein The immunomodulatory agent or warhead of the provided compound, when conjugated (e.g., targeted conjugate) It becomes active (when conjugated to a gate). In some embodiments, nuclear head / immune While the regulatory substance is active, the non-release linker and targeting moiety (for example, in the body of the target) Therefore, toxic cytokines that activate the immune system (for example, interleukin-6 (IL-6) ) and so on) prevent the release of (for example, (using folic acid or its analogues) (Because the substance is specifically targeted). In certain cases, for example, a compound warhead / immunomodulation. Even if a substance is active when it is connected to a non-release linker, immunomodulators are compounds that are targeted Until it binds to the target receptor (for example, the folate receptor), it will need to be properly located within the cell's endosomes (for example) If this happens, the target receptor cannot be accessed.

[0128] As a non-limiting example, linker 106 in Figure 1A is a non-release PEG linker, The linker 156 in Figure 1B is a self-immolative releaseable linker. It is a linker (for example, containing a disulfide bond (e.g., SS)). For example, Figure 1B This shows the self-destructive cascade of compound 150 upon cleavage from the targeted portion 154.

[0129] In some embodiments, the linker 156 is used to ensure that the compound circulates within the target systemic circulation after administration. After sufficient time has elapsed (for example, after it has disappeared from non-target tissue and the target cells and / (or only when captured and internally transferred by the receptor) the drug is delivered from the targeting portion 154 It is formed to be cut. In some embodiments, the duration of release varies (for example) For example, depending on the target (based on various factors). In some embodiments, release is possible. The linker can be cut off for at least 24 hours after administration, or even up to a week later. They may be manipulated to prevent release. In some embodiments, the compound is used to control the system in question. It can pass through safely and be captured by target cells (e.g., those expressing FRβ). Any amount that is not released can be purged before release / activation, thus preventing toxicity (e.g.) For example, when bound to a releaseable linker, the immunomodulator is not active.

[0130] Both releaseable and non-releaseable linkers affect the in vivo distribution (of compounds, for example), biological distribution. Optimize usability and PK / PD, and / or PEGlation (PEGlay) Methodologies that are generally known or will be developed in the field through (tion), etc. To manipulate (for example, a compound) to increase its uptake into the target tissue. This can be done. In some embodiments, the linker is a cell (for example, a target cell (e.g.) If, before being captured by macrophages in the fibrous or cancerous tissue being treated, It is configured to avoid a significant release of the pharmaceutical activity of the drug.

[0131] In some embodiments, compounds comprising the releaseable linker of the present disclosure are used in endosomes. They can be designed to diffuse across membranes, for example, into the cytoplasm of target cells. In this embodiment, the releaseable linker releases immunomodulatory substances until the compound reaches the cytoplasm. It can be designed to prevent this from happening.

[0132] In some embodiments, the conjugate provided herein is an ejectable linker (For example, to promote the release of immunomodulatory substances in the cytoplasm, for example, in that case, immunomodulatory The nodal substance is responsible for the activity of PI3K kinase, IRAK, or 1-kappa-β(Iκβ) kinase. Activation factors (e.g., using prostratin) or nuclear factor kappa in activated B cells - Light chain enhancer (NF-κβ) (see Table 1, for example), or myelodifferentiation Includes the next response 88 (MyD88) agonist. In some embodiments, it is emitting A linker, for example, has a targeting portion that targets the appropriate target (e.g., macrophage folate receptor). It binds and is internally transported into the endosomes of target cells and / or into the cytoplasm (e.g., The release of immunomodulatory substances is delayed until they have diffused to the location where the desired pattern recognition receptor is located. To prevent. In some embodiments, the releaseable linker delivers immunomodulatory substances into the endosome. Release quality.

[0133] In some embodiments, the linker provided herein has one or more spacers ( For example, to promote a specific release time, to promote increased uptake into target tissue, and / or the in vivo distribution, bioavailability, and / or the compounds provided herein. The spacer may include alkyl chains, PEG, and PEG (to optimize PK / PD). Tides, sugars, peptidoglycans, clickable linkers (e.g., triazoles), poly One of the hard linkers, such as proline and polypiperidine, and similar substances. It may include one or more.

[0134] In some embodiments, PEG 12 Linkers containing the compounds provided herein Nonspecific uptake (e.g., non-targeted organs (e.g., the liver and / or the target after administration) If the amount of blood entering the kidney is not completely avoided, it will be significantly reduced. Therefore, the compound avoids delivery to the liver and kidneys. In some embodiments, it is targeted. The part (its free form, its radical, or its conjugate) on non-targeted cells It does not bind to the uptake receptor (for example, if the organ is not the target site, therefore it Stimulation of immune complexes in these organs can be avoided, which is non in clinical situations. (Always beneficial).

[0135] In some embodiments, a conjugate including a non-dischargeable linker provided herein is used. The free form (for example, the free form of the compounds and / or ligands provided herein) (In a separate form) it reduces or eliminates the toxicity of components released from the conjugate.

[0136] In at least one embodiment, the linker includes a hydrophilic spacer. Several embodiments Morphologically, the compound has the structure of formula XII (e.g., release containing the first hydrophilic spacer). Sub-sub It has a structure. [ka]

[0137] In some embodiments, the compound has a structure of formula XIII (for example, a second hydrophilic space) Formula III is conjugated with folic acid via a non-release linker (covalent bond) containing sir. It has a substructure of the TLR7 agonist. [ka]

[0138] Specific examples of exemplary conjugate compounds are provided herein.

[0139] In some embodiments, the compounds provided herein are immunomodulators (or their equivalents). When conjugated with a radical or a pharmaceutically acceptable salt thereof, it becomes pharmaceutically active. It remains conjugated with radicals of immunomodulators or their pharmaceutically acceptable salts. The targeted portion contains radicals of the targeted portion as described herein. It may include a portion, and in at least one embodiment, a folate ligand, any other folate receptor Container-binding molecules (for example, or functional fragments or analogues of any of the above) or includes pyrido[2,3-d]pyrimidine analogs. In some embodiments, targeted The portion (or its conjugate or radical) is specific to FRβ.

[0140] In some embodiments, the compounds provided herein include one or more linkers. The radical of the targeted portion is converted to an immunomodulatory radical via one or more linkers. It is jugated. For example, an immunomodulator or a pharmaceutically acceptable salt thereof is of formula I or If II is present, the radical of the immunomodulatory substance is transmitted via the linker or directly to R 1 , R 2 , or R 3 In one of them, the radical of the targeting part is conjugated It may also be said that an immunomodulator or a pharmaceutically acceptable salt thereof has formula III. In this case, the radical of the immunomodulator is transmitted via the linker or directly, R 1 or R 3 In one of these, the radical of the targeting portion may be conjugated. If the immunomodulator or a pharmaceutically acceptable salt thereof has formula IV, the immunomodulator Radicals, either via the linker or directly, R 1 or R 2 In one of them , may be conjugated to the radical of the targeting portion. As described herein, The linker may be releaseable or not.

[0141] In some embodiments, one or more linkers of the compounds provided herein are PE G, PEG derivatives, or known in the art that can achieve the purposes specified herein Or it may include any other linker to be developed in the future. In some embodiments, The check can be repeated n times, and n is a positive integer. For example, n is not limited to The answer is any integer selected from the ranges 1-16, 1-32, 1-64, or 1-96. This may also be the case. The number of repeats in the linker (i.e., n) is determined by the desired functionality of the compound. Selected considering the is and / or the desired application in order to achieve the effect and / or to achieve the desired effect. In some embodiments, one or more of the linkers may be one or more space Includes a ser (for example, this can also be used to specifically design the properties of a compound). .

[0142] In some embodiments, the linker is a hydrolyzable linker. In this state, the linker is a non-hydrolyzable linker. In some embodiments, the linker is an optionally substituted heteroalkyl. In some embodiments, the linker It consists of alkyl, hydroxyl, oxo, PEG, carboxylate, and halo. It is a substituted heteroalkyl group containing at least one substituent selected from the group. In the embodiment, the linker is a spacer (for example, as described elsewhere in this specification) Includes.

[0143] In some embodiments, the linker has at least one disulfide bond in its framework It is a substituted heteroalkyl having a compound. In some embodiments, the linker is its skeleton It is a peptide that has at least one disulfide bond inside.

[0144] In some embodiments, the linker is -CONH-CH(COOH)-CH2-S- S-CH2-CR a R b -O-CO-, -CONH-CH(COOH)CR a R b -O- CO-, C(O)NHCH(COOH)(CH2)2-CONH-CH(COOH)CR a R b -O-CO- or -C(O)NHCH(COOH)(CH2)2-CONH-C H(COOH)-CH2-SS-CH2-CR a R b -O-CO- is included in the formula, where R a and R b These are independently H, alkyl, or heteroalkyl (e.g., PEG). .

[0145] In some embodiments, the linker includes the following structure: [ka] In the formula, n and m are independently between 0 and 10.

[0146] In some embodiments, the linker includes the following structure: [ka] In the formula, n and m are independently between 0 and 10.

[0147] In some embodiments, the linker includes the following structure: [ka] In the formula, n is between 1 and 32.

[0148] In some embodiments, the linker includes the following structure: [ka] In the formula, n is between 1 and 16.

[0149] The radical of a compound (for example, the radical of a compound in either Table 1 or Table 2) Radicals of ligands, linkers (e.g., those provided herein), and ligand radicals. Any combination of ligand radicals (for example, any one of the radicals in Tables 3-6) By combining them, the conjugates provided herein can be formed. This is understood. In some embodiments, the radical of the compound or the radical of the ligand is , carbon atoms or heteroatoms (e.g., O, S, N, etc.). In some embodiments The radical of the compound is C or O. In some embodiments, the radical of the ligand is C. Cal is C or O. In some embodiments, the attachment site of the compound and ligand ( For example, via a linker, the arrangement is determined by the arrangement of radicals. Several implementations In this configuration, the linker includes spacers (for example, those described elsewhere in this specification). Any conjugate provided herein is provided in the methods provided in the examples. It is also understood that it can be synthesized using a similar process.

[0150] Non-limiting examples of conjugates provided herein are provided in Table 7. [Table 7] TIFF2026090286000080.tif244170TIFF2026090286000081.tif91170

[0151] Non-limiting examples of conjugates provided herein are provided in Table 8. [Table 8] JPEG2026090286000083.jpg254170JPEG2026090286000084.jpg230170TIFF2026090286000085.tif251170TIFF2026090286000086.tif207170

[0152] In some cases, the conjugate compounds provided herein have a structure of formula XIV (e.g. For example, or a functional fragment or analog thereof, folic acid and condyloma via a releaseable linker. (Containing a TLR7 agonist of the eugated formula III): [ka]

[0153] In another embodiment, the conjugate compound provided herein has a structure of formula XV (e.g. For example, or a functional fragment or analog thereof, folic acid and condyloma via a releaseable linker. The compound has a fused formula II TLR7 agonist (e.g., compound 3B): [ka]

[0154] In yet another embodiment, the conjugate compound provided herein is of formula XVI Structure (for example, or a functional fragment or analog thereof, containing three PEGs, non-release lignes) Contains a TLR7 agonist of formula II conjugated with folic acid via a linker (e.g.) , has compound 3D)): [ka]

[0155] In yet another embodiment, the conjugate compound provided herein is of formula XVII The structure (for example, or a functional fragment or analogue thereof, containing 12 PEGs) Includes a TLR7 agonist of formula II conjugated with folic acid via a polylinker (e.g.) For example, having compound 3C): [ka]

[0156] Further embodiments of the conjugate compounds provided herein are of the composition of formula XVIII. (For example, a functional fragment or analog thereof containing 16 PEGs, which cannot be released) Contains a TLR7 agonist of formula II conjugated with folic acid via a linker (e.g.) , containing compound 3D'): [ka]

[0157] Further embodiments of the conjugate compounds provided herein have the structure of formula XIX ( For example, formula III conjugated with folic acid, or a functional fragment or analog thereof. It contains a TLR7 agonist (compound 1B): [ka]

[0158] The compounds described herein are derived from conventional organic synthesis methods practiced by those skilled in the art. Therefore, it can be prepared. The general reaction sequence outlined below is for preparing the compounds of this disclosure. This represents a general method that is useful and does not mean to limit its scope or usefulness. .

[0159] The description of the compounds of the present invention is limited by the principles of chemical bonding known to those skilled in the art. However, In cases where a group can be substituted by one or more substituents, such a configuration The exchange is based on the principles of chemical bonding, and the compounds are not inherently unstable, and / or aqueous. It is likely to be unstable under ambient conditions such as neutrality and some known physiological conditions. A compound is selected to give a compound that is well known to those skilled in the art. For example, heterozygous Chloalkyl or heteroaryl compounds are formed according to the principles of chemical bonding known to those skilled in the art, and are cyclic heteroaryl. It attaches to the rest of the molecule via atoms, thereby avoiding the inherently unstable compound. ru.

[0160] The terms “identical” or “identity” relating to two or more polypeptide sequences are relevant. Using array comparison algorithms known in the art or by manual alignment and visual inspection When measured by the test, they are either identical or a specific percentage of the same peptide. (i.e., comparison window or targeting end, folate end, linker, or warhead, etc.) When compared and aligned for the greatest correspondence across the specified area, the specified Approximately 60% identity across the selected area, preferably 65%, 70%, 75%, 80%, 8%. 5%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 9 This refers to two or more sequences or subsequences that have 9% or higher identity. In such cases, such sequences are said to be "substantially identical." In other words, one of the molecules The general shape and structure, and where appropriate, the hydrogen bonds, are substantially adapted to the target binding site. And, as long as it is maintained to function as an agonist against it, one or more of the entire sequence Identity exists within that domain.

[0161] The compounds described herein include one or more pharmaceutically acceptable carriers, adjuvants, Unit dosage forms including diluents, excipients, and / or vehicles, and combinations thereof. It may be administered in and / or composition. When used herein, the term "administration" "and its formations can be administered orally, intravenously, intramuscularly, subcutaneously, transdermally, by inhalation, through the mouth, eyes, sublingually, orally vaginally. The compounds described herein include, but are not limited to, rectal and similar routes of administration. This generally refers to any means of introducing a substance into a host.

[0162] The administration of the compounds of this disclosure as salts may be appropriate. Examples of acceptable salts include, but are not limited to, However, alkali metals (e.g., sodium, potassium, or lithium) or alkaline It contains earth metal (e.g., calcium) salts, however, when administered to the subject being treated... Any salt that is generally non-toxic and effective is acceptable when used. Similarly, "pharmacopoeiatically acceptable "Salt" refers to a salt that has counterions and may be used in pharmaceuticals. Such salts are not limited to The following may be included: (1) an acid addition salt, which is a free base of the parent compound and an inorganic acid, e.g. For example, through reactions with hydrochloric acid, hydrobromic acid, nitric acid, phosphoric acid, sulfuric acid, and perchloric acid, Or organic acids, such as acetic acid, oxalic acid, (D) or (L) malic acid, maleic acid, meta Sulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, tartaric acid, quer It can be obtained by reaction with nic acid, succinic acid or malonic acid, or (2) a herbaceous Acidic protons present in the compound can be replaced by metal ions, such as alkali metal ions and alkaline earth ions. It is substituted with an ion-like substance or an aluminum ion, or with an organic base, such as ethanol. Diethanolamine, triethanolamine, trimethamine, N-methyl methylamine Salts formed when coordinated with lucamin, etc. Pharmacologically acceptable salts are: Any such pharmaceutically acceptable salts, which are well known to those skilled in the art, are described herein. This may be intended in relation to the embodiments described.

[0163] An acceptable salt is obtained by reacting a sufficiently acidic compound with a suitable base to produce a physiologically acceptable salt. Standard procedures known in the art, including (but not limited to) providing anions. It can be obtained using [method]. A suitable acid addition salt is formed from an acid that forms a non-toxic salt. Example Exemplary and non-exclusive examples include acetates, aspartates, benzoates, besilates, and heavy salts. Carbonates / carbonates, bicarbonates / sulfates, borates, camsilates, citrates, edicylates Salt, esylate, formate, fumarate, gluceptate, gluconate, glucuronide Hexafluorophosphate, hibenzate, hydrochloride / chloride, hydrobromide / bromide, yo Hydrochloride / iodide, isethionate, lactate, malate, maleate, malon Salts, mesylates, methylsulfates, naphthylates, 2-nap Silates, nicotinates, nitrates, orotates, oxalates, palmitates, pamoic acid Salts, phosphates / hydrogen phosphates / dihydrogen phosphates, sugars, stearates, succinates, This includes tartrates, tosylates, and trifluoroacetates. A suitable base salt is formed from a base that forms a non-toxic salt. (Examples are illustrative and non-limiting.) It contains arginine, benzathine, calcium, choline, diethylamine, diolamine, and glycan. Syn, lysine, magnesium, meglumine, olamine, potassium, sodium, trometa Examples include, but are not limited to, mine and zinc salts. Hemi salts of acids and bases (h Emisalts, for example, hemisulfate salts and hemicalcium salts can also be formed.

[0164] As used herein, the term “composition” includes the compounds described herein. This generally refers to any product containing two or more components. The compositions described herein are... From isolated compounds described in the specification, or salts of compounds described herein, soluble It should be understood that it can be prepared from liquids, hydrates, solvates, and other forms. Certain functional groups, such as hydroxyl, amino, and similar groups, can influence the various physical forms of a compound. It is understood that it can form complexes with water and / or various solvents. The composition is The various amorphous, amorphous, partially crystalline, and crystalline properties of the compounds described in the specification. and / or other morphological forms may be prepared, and the composition is a compound as described herein. It should also be understood that it can be prepared from various hydrates and / or solvates of the substance. Therefore, pharmaceutical compositions that list the compounds described herein include those described herein. The various morphological forms and / or solvated or hydrated forms of the compound being treated, This includes any combination of these, or individual forms thereof.

[0165] The compounds disclosed herein are formulated as pharmaceutical compositions and are adapted to various routes of administration. It can be administered to mammalian hosts such as human patients in various forms. For example, the pharmaceutical composition may be administered orally or This includes parenteral, intravenous, intraarterial, intraperitoneal, subarachnoid, epidural, intraventricular, urethral, ​​and intrasternal administration. Formulated for intracranial, intratumor, intramuscular, topical, inhalation, and / or subcutaneous routes, They can be administered via them. In fact, in at least one embodiment described herein The listed compounds and / or compositions may be administered directly into the bloodstream, muscle, or viscera. good.

[0166] For example, in at least one embodiment, the compound is an inert diluent or an assimilated food. In combination with a pharmaceutically acceptable vehicle such as a carrier, it can be administered systemically (for example, orally). ) may be used. For oral therapeutic administration, the active compound may be combined with one or more excipients. These are ingestible tablets, buccal tablets, lozenges, capsules, elixirs, suspensions, and syrups. It can be used in the form of chips, wafers, etc. The percentages of the composition and preparations may vary. It may be done in a quantity of approximately 1 to 99% by weight, consisting of an active ingredient, binder, excipient, disintegrant, lubricant, etc. It may be between and / or sweeteners (known in the art). Such therapeutic The amount of the active compound in the composition useful for this purpose is such that an effective dose level can be obtained.

[0167] For example, the preparation of parenteral compounds / compositions under sterile conditions by freeze-drying is well known to those skilled in the art. This can be easily achieved using standard pharmaceutical technology. In at least one embodiment, non The solubility of compounds used in the preparation of oral compositions is determined by solubility enhancers. By using appropriate formulation techniques such as incorporating an enhancing agent It can be increased.

[0168] As previously stated, the compounds / compositions of this disclosure may also be administered by injection or injection (for example, Administered using needle syringes (including microneedles) and / or needleless syringes. The solution of the active composition may be aqueous and optionally mixed with a non-toxic surfactant. They may be combined and / or used as carriers or excipients such as salts, carbohydrates, and buffers. It may contain (preferably at a pH of 3 to 9), but for some applications, These can be used as a sterile non-aqueous solution, or with sterile pyrogen-free water or phosphate-buffered water. It is more appropriately formulated as a dry form to be used with a suitable vehicle such as saline solution (PBS). It may be modified. For example, the dispersion may contain glycerol, liquid PEG, triacetin, and These mixtures, as well as those in oil, can be prepared under normal storage and use conditions. These preparations may further contain preservatives to prevent the growth of microorganisms.

[0169] Suitable drug dosage forms for injection or infusion are optionally liposome-encapsulated, sterile, and injectable. A sterile aqueous solution or a dispersant containing an active ingredient suitable for immediate preparation. It may contain a dispersant or sterile powder. In all cases, the final dosage form is determined by the manufacturing process. It should be sterilized, fluid, and stable under the conditions of storage. Liquid carrier or liquid via The ingredients include, for example, water, ethanol, and polyols (e.g., glycerol). , propylene glycol, liquid PEG, etc.), vegetable oil, non-toxic glyceryl ester, and It may be a solvent or liquid dispersion medium containing and / or a suitable mixture thereof. In another embodiment, adequate fluidity is achieved by the formation of liposomes, which is necessary in the case of a dispersion. This can be maintained by maintaining the required particle size or by using surfactants. The microbial action is parabens, chlorobutanol, phenol, sorbic acid, thimerosulfame This can be prevented by adding various antibacterial and antifungal agents such as . It is desirable that the solution contains one or more isotonic agents, such as sugar, buffer solution, or sodium chloride. The long-term absorption of injectable compositions may be slowed down by the formulation of drugs, for example, This can be achieved by incorporating aluminum monostearate and gelatin.

[0170] Sterile injectable solutions contain active compounds and / or compositions as specified above, if necessary. Combine one or more of the other components with the required amount of appropriate solvent, and then filter. It can be prepared by sterilization. In the case of sterile powders for the preparation of sterile injectable solutions, Preferred preparation methods are vacuum drying and freeze-drying techniques, which involve the powdering of the active ingredient. This results in a solution that has been previously sterile filtered and then mixed with any additional desired components. .

[0171] For topical administration, the compound is administered to a dermatologically acceptable carrier, which may be solid or liquid. It may be desirable to administer it to the skin as a composition or formulation in combination with other substances. For example In certain embodiments, the solid carrier may be talc, clay, microcrystalline cellulose, silica, aluminum, etc. It may contain finely divided solids such as mina. Similarly, useful liquid carriers include water, alcohol, etc. This may contain alcohol or glycol, or a water-alcohol / glycol blend. The compound of the invention is optionally dissolved to an effective level with the help of a non-toxic surfactant. It can be dispersed. Additionally or alternatively, adjuvants such as fragrances and antimicrobial agents can be added. By adding a bunt, the properties for a given use can be optimized. The resulting liquid composition The material is applied from an absorbent pad or impregnated with bandages and / or other bandages. Use it, or spray it onto the target area using a pump or aerosol sprayer, or Alternatively, it can simply be applied directly to the desired area of ​​the target.

[0172] Synthetic polymers, fatty acids, fatty acid salts and esters, fatty acid alcohols, modified cellulose Alternatively, a thickening agent such as a modified mineral is used together with a liquid carrier and applied directly to the target skin. To achieve this, it can also form pastes, gels, ointments, soaps, etc., that can be spread.

[0173] As used herein, the terms "therapeutably effective," "therapeutic effective dose," and "therapeutic amount" are used. The terms "protective dose" or "protective dose" are (unless otherwise specified) If administered once or as part of a treatment cycle, the target health and well-being This refers to the amount of a compound that affects the stimulant or mortality rate (for example, not limited to...) However, if applicable, one or more related to fibrous disease or condition and / or cancer. (To delay the onset of symptoms and / or reduce their severity). Useful applications of the compounds of this disclosure The quantity is used to compare their in vitro and in vivo activities in animal models. This can be determined by the effective dose in mice and other animals and its application to humans. The method of extrapolation is known in the art. In practice, the dosage of the compound depends on the host subject's condition. The type of cancer or fibrous disease being treated, the stage of the disease progression, and the route of compound administration. and tissue distribution, as well as other therapeutic treatments (additional drugs in radiotherapy or combination therapy). Depending on the possibility of concomitant use of treatments (e.g., effective treatment or prevention), the results may vary significantly. The amount of composition required for use in a given quantity or dose depends not only on the specific application but also on the selection. The salt (if applicable) and the characteristics of the subject (e.g., age, condition, sex, body surface of the subject) It also varies depending on the product and / or mass, drug resistance, etc., and ultimately, the attending physician It is left to the discretion of the doctor, clinician, or other person. The therapeutically effective or prophylactic effective dose or dosage. The dosage is, for example, approximately 0.05 mg / kg patient weight to approximately 30.0 mg / kg patient weight, or The range may be approximately 0.01 mg / kg patient body weight to approximately 5.0 mg / kg patient body weight, and is limited. It is not that, but 0.01 mg / kg, 0.02 mg / kg, 0.03 mg / kg, 0 .04mg / kg, 0.05mg / kg, 0.1mg / kg, 0.2mg / kg, 0.3 mg / kg, 0.4mg / kg, 0.5mg / kg, 1.0mg / kg, 1.5mg / k g, 2.0mg / kg, 2.5mg / kg, 3.0mg / kg, 3.5mg / kg, 4. 0 mg / kg, 4.5 mg / kg, and 5.0 mg / kg (all in kg of the patient's body weight) It contains. The total therapeutic or prophylactic effective dose of the compound is administered in single or divided doses. This may also be outside the typical scope given herein, at the discretion of the practicing physician.

[0174] In another embodiment, the compound is present in concentrations of approximately 0.5 g / m to approximately 500 mg / m. 2 , about 0.5g / m 2 ~about 300mg / m 2 , or approximately 100g / m 2 ~about 200mg / m 2 Treatment or prognosis It may be administered in an effective dose. In other embodiments, the dose is about 0.5 mg / m². 2 ~about 500mg / m 2 , about 0.5mg / m 2 ~about 300mg / m 2 , about 0.5mg / m 2 ~about 200mg / m 2 , about 0.5mg / m 2 ~about 100mg / m 2 , about 0.5mg / m 2 ~about 50mg / m 2 , about 0.5mg / m 2 ~about 600mg / m 2 , about 0.5mg / m 2 ~6.0mg / m 2 , about 0.5mg / m 2 ~about 4.0mg / m 2 , or approximately 0.5 mg / m² 2 ~about 2.0 mg / m 2 This may be the case. The total amount may be administered as a single dose or in divided doses, at the discretion of the physician. These quantities may be outside the typical range given herein. Based on.

[0175] In some embodiments, the expression of a specific biomarker in a sample from a subject is measured. In relation to the analysis of cytokine levels, the significance of this disclosure lies in the markers or Not a specific method used to detect a set of markers, but rather detecting markers It is used for the expression, quantification, or profiling of one or more biomarkers. There are many methods that can be used to detect the virus. Alternatively, once a set of markers is identified, along with the provision of appropriate reagents, several techniques (current Either known or future development methods may be used. If one or more biomarkers are provided to be identified, the method disclosed herein An appropriate assay to perform (e.g., PCR-based or my assay for nucleic acid markers) Cross assay-based assays, enzyme-linked immunosorbent assays (ELISA), proteins Alternatively, you can choose an antibody microarray or a similar immunological assay. ru.

[0176] Chemical examples Example A: Synthesis of Compound 1A: Compound 1A was prepared according to Scheme 1 below by Nikunj M. Shukla, Col. e A. Mutz, Subbalakshmi S. Malladi, Hemamli J.Warshakoon,Rajalakshmi Balakrishna,and Sunil A. David, “Regioisomerism-dependent TLR7 agonism and antagonism in an imida zoquinoline;Structure-Activity Relations hips in Human Toll-Like Receptor 7-Active e Imidazoquinoline Analogues,”J Med Chem As reported by .2012 Feb 9;55(3):1106-1116 I did it. [ka]

[0177] Step 1: Synthesis of l-amino-2-methylpropan-2-ol (compound) 2,2-dimethyloxirane (0.1 g, 1.388 mmol) is chilled in 20 mL of ice. It was added dropwise to a solution of ammonium hydroxide. The reaction mixture was stirred at room temperature for 12 hours. The solvent was removed under vacuum, and the residue was dissolved in methanol. Di-tert-butyl dicarbonate (0 Add 0.75g (3.47 mmol) to the reaction mixture and stir for 4 hours. The mixture was then subjected to column chromatography. Purified using chromatography (24% ethyl(SiO) / hexane) tert-butyl 2-hydroxy-2-methylpropylcarbamate was obtained. Pure t Add ert-butyl 2-hydroxy-2-methylpropyl carbamate to 5 mL of Triflu Dissolve in oloacetic acid and stir for 35 minutes. Remove solvent under reduced pressure and extract trifluoroacetate 1' l-amino-2-methylpropan-2-ol was obtained as the compound. 1H NMR 500MH z(500MHz, CDC13, δ(ppm)):δ 8.62(s,2H),3.02 (d,2H),2.06-2.04(m,2H),1.37-1.34(s,6H).

[0178] Step 2: 2-methyl-l-(3-nitroquinoline-4-ylamino)propane-2- Synthesis of ol (compound 2) l-amino-2-methylpropan-2-ol (compound) (450 mg, 2.4 mmol) l) Add the trifluoroacetate in a 4:1 mixture of toluene and 2-propanol. -Chloro-3-nitroquinoline (compound 1) (250 mg, 1.2 mmol) and Et It was added to a 3N (0.5 ml, 3 mmol) solution. The mixture was stirred until the solid began to precipitate. The mixture was heated to 70°C for 30 minutes. The reaction mixture was then cooled, filtered, and toluene / 2-Pluginseng was removed. Washed with ropanol (7:3), ether, and cold water. Dry the residue at 80°C. -Methyl-l-(3-nitroquinoline-4-ylamino)propan-2-ol (compound) 2) was obtained. Liquid chromatography-mass spectrometry (LCMS) analysis: [M+H] + m / z = 261.

[0179] Step 3: l-(3-aminoquinoline-4-ylamino)-2-methylpropane-2- Synthesis of ol (compound 3) 2-methyl-l-(3-nitroquinoline-4-ylamino)propan-2-ol (chemical Mixture 2) (450 mg, 1.72 mmol) is dissolved in methanol, and Pd / C is used as a catalyst. The solution was then hydrogenated using a hydrogen balloon for 4 hours. Next, the solution was filtered using Celite. Then, the solvent is evaporated under reduced pressure to obtain l-(3-aminoquinoline-4-ylamino) -2-ethylpropane-2-ol (compound 3) was obtained. LCMS:[M+H] + m / z=231.Η NMR 500 MHz(CDC13, δ(ppm)):δ 8.12 (s,1H),7.61-7.58(m,1H),7.48-7.40(m,2H),4 .90(s,2H),3.47(2H),1.35-1.21(s,6H).

[0180] Step 4: l-(4-amino-2-butyl-lH-imidazo[4,5-c]quinoline- Synthesis of l-yl)-2-methylpropan-2-ol (compound 5, TLR7A) A solution of compound 3 (100 mg, 0.43 mmol) in anhydrous THF is mixed with triethylamine. (66 mg, 0.65 mmol) and valeryl chloride (62 mg, 0.52 mmol) The solvent was then added. The reaction mixture was stirred for 6-8 hours, followed by the removal of the solvent under vacuum. The residue was dissolved in siRNA, washed with water and brine, and then dried with Na2SiO4. This yielded an intermediate amide compound. This was dissolved in methanol (MeOH) and then oxidized. Calcium was added and heated in a microwave at 110°C for 1 hour. Then the solvent was removed. The residue was purified using column chromatography (9% MeOH / dichloromethane). Compound 4 (58 mg) was obtained. MeOH:dichloromethane:chloroform (0.1: A solution of compound 4 in a 1:1 solvent mixture is prepared by adding 3-chloroperoxybenzoic acid (84 mg). Add (0.49 mmol) and reflux the solution at 45-50°C for 40 minutes. Then remove the solvent. Remove the residue and use column chromatography (20% MeOH / dichloromethane) The oxide derivative (55 mg) was obtained by purification. This was then dissolved in anhydrous dichloromethane. Next, benzoyl isocyanate (39 mg, 0.26 mmol) was added, and the mixture was heated at 45°C. It was heated for 15 minutes. Then, the solvent was removed under vacuum, and the residue was dissolved in anhydrous MeOH, and continued... Then, an excess of sodium methoxide was added. The reaction mixture was then heated at 80°C for 1 hour. The mixture was heated. The solvent was removed under vacuum, and the residue was subjected to column chromatography (11% MeOH / di). Compound 5 was obtained by purification using chloromethane. LCMS:[M+H] + m / z = 312. H NMR 500MHz(CDC13, δ(ppm)):δ 8.16-8. 15(d,1H),7.77-7.46(d,1H),7.46-7.43(m,1H) ,7.33-7.26(m,1H),3.00-2.97(m,2H),1.84-1. 78(m,2H),1.47-1.41(m,2H),1.36(s,6H),0.98 -0.95 (m, 3H).

[0181] Example B: Synthesis of Compound 1B: Subsequently, compound 1B is synthesized using compound 1A according to scheme 2 below. It is possible. [ka]

[0182] Compound 1A, folic acid, and linker are commercially available or can be manufactured by methods known to those skilled in the art. Therefore, it can be prepared.

[0183] Heterobifunctional linker 7 (88 mg, 0.213 mmol) was subjected to a nitrogen atmosphere at room temperature. Compound 5 (33 mg, 0.106 mmol) and dimethyl in 4 mL of methylene chloride. Add the aminopyridine (39 mg, 0.319 mmol) solution to the mixture and heat at reflux temperature for 7 minutes. Stirring for a certain amount of time, during which thin-layer chromatography (TLC) analysis of the mixture showed a change of >80%. The mixture was concentrated, and 10% acetonitrile in methylene chloride was used as the eluent. The product was purified by column chromatography. The pure product compound 9 was obtained as a pale yellow solid. It was obtained by doing so. A solution of compound 8 (1 equivalent) in dimethyl sulfoxide (DMSO) was prepared in dimethyl sulfoxide (DMSO). Drug linker intermediate compound 9 (1.0 equivalent) in DMSO containing 1 equivalent of 1 aminopyridine The solution (approximately 1.5 equivalents) was added in three separate doses at 20-minute intervals. Under argon, at room temperature, 1 After stirring for approximately 2 hours, LC-MS analysis of the mixture revealed the desired folic acid-drug compound as the main product. The formation of jugate (compound 10) was observed. The mixture was subjected to preparative high-performance liquid chromatography. Purified by HPLC. LCMS:[M+H] + m / z = 959. 1 1H NMR (500 MHz,DMSO-d6)δ 8.58(s,1H),8.49(d,J=8 .8 Hz,1H),7.90(d,J=8.3 Hz,1H),7.83-7.74( m,1H),7.54(d,J=8.0 Hz,2H),7.48(t,J=7.6H z,1H),7.41(s,1H),7.06(s,1H),6.81(d,J=6.2 Hz,1H),6.61(d,J=8.3 Hz,2H),6.27(s,1H),4 .43(d,J=5.9 Hz,2H),4.28(t,J=6.6 Hz,2H),4 .00(d,J=25.7 Hz,3H),3.03(t,J=7.5 Hz,2H), 2.97(dd,J=13.0,6.5Hz,1H),2.09(s,2H),1.8 1(s,7H),1.40(q,J=7.4Hz,2H),1.22(s,2H),1 .13(s,2H),0.91(t,J=7.4 Hz,3H).

[0184] Example C: Synthesis of Compound 2A: Compound 2A can be synthesized according to schemes 3 and 4. [ka] [ka]

[0185] Wang resin (11) loaded with cysteine ​​in dimethylformamide (DMF) The free amine was first deprotected using 20% ​​piperidine. 1-Il-oxytripyrrolidinophosphonium hexafluorophosphate (PyBop ), in the presence of N,N-diisopropylethylamine (DIPEA) and DMF, Fm The product was treated with oc-Glu(OtBu)-COOH. The bound product was then added to 20% of the DMF. Deprotection was performed using piperidine, and in the presence of PyBop, DIPEA, and DMF, pterodactyl The compound 12 was produced by treatment with leucoic acid. The trifluoroacetyl group was removed with 50% ammonia. -The resin was deprotected with DMF solution. Finally, the resin was treated with trifluoroacetic acid:triisopropyl silamine. Cut using a cocktail solution of water and tris(2-carboxyethyl)phosphine, H Folic acid-cysteine ​​(13) was obtained as a yellow solid by purification using PLC.

[0186] Compound 14 was first treated with a heterobifunctional linker reagent (15) to folate-cystic acid A disulfide intermediate (16) was obtained. This was then mixed with folate-cysteine ​​in DMSO ( 13) is reacted with and purified using HPLC to produce compound 17 (e.g., compound 2A). The characterization of all compounds was performed using ammonium bicarbonate and acetonite as buffer systems. LC-MS using Trill was employed. The mass observed from LC-MS of compound 2A was [M +H]+=1082.2

[0187] In particular, considering schemes 1, 2, and 3 specified above, the current state of the related technologies and Furthermore, this disclosure will allow those skilled in the art to utilize the concepts expressed herein to understand all other aspects of this disclosure. It provides sufficient detail to enable the synthesis of the compound.

[0188] Treatment method In addition to the compounds described herein, the treatment and / or prevention of fibrous diseases or cancer. Methods for prevention will also be provided.

[0189] In some embodiments, provided herein are cases of fibrotic disease or cancer. A method for treating a diseased subject, the method comprising the cells of the subject, provided herein Any compound, or a pharmaceutically acceptable salt thereof, or any of the compounds provided herein. Contact with a compound or a pharmaceutically acceptable salt thereof in a (e.g., pharmaceutical) composition. A method that includes, for example, a compound, formula I, formula II, formula III, formula IV, formula V, Formula VI, Formula VII, Formula VIII, Formula IX, Formula X, Formula XI, Formula XII, Formula XIII, Formula XI V, Formula XV, Formula XVI, Formula XVII, Formula XVIII, Formula XX, or Formula XXX, Household Agency It has one structure of intent. In some embodiments, the immunomodulator is TLR7, 8, This includes 9 agonists.

[0190] In some embodiments, the following is provided herein: A method for treating inflammatory, autoimmune, or fibrous diseases or disorders, as specified herein. Any compound provided to this, or a pharmaceutically acceptable salt thereof, or provided herein A composition (e.g., a pharmaceutical composition) containing any compound or a pharmaceutically acceptable salt thereof. A method comprising administering a substance to an individual, for example, a compound, formula I, formula II, formula III, Formula IV, Formula V, Formula VI, Formula VII, Formula VIII, Formula IX, Formula X, Formula XI, Formula XI I, Formula XIII, Formula XIV, Formula XV, Formula XVI, Formula XVII, Formula XVIII, Formula XX, It has any one structure from among the formulas XXX.

[0191] In some embodiments, the following is provided herein: A method for treating a disease or disorder, comprising any compound provided herein, or The pharmaceutically acceptable salt thereof, or any compound provided herein, or the drug thereof. This includes administering to the individual a composition (e.g., a pharmaceutical) containing a scientifically acceptable salt. , a method, for example, compounds are formulas I, II, III, IV, V, VI, V II, Formula VIII, Formula IX, Formula X, Formula XI, Formula XII, Formula XIII, Formula XIV, Formula XV, Any one of the following formulas: formula XVI, formula XVII, formula XVIII, formula XX, or formula XXX It has a structure. In some embodiments, the neoplastic disease or disorder is cancer. In this embodiment, cancer includes bladder cancer, brain cancer, liver cancer, kidney cancer, skin cancer, thymic cancer, and The cancers selected are endoplasmic stromal tumors (GIST), esophageal cancer, pancreatic cancer, and breast cancer.

[0192] In some embodiments, the fibrous condition of an individual requiring treatment is provided herein. A method for treating a disease or disorder, comprising any compound provided herein, or The pharmaceutically acceptable salt thereof, or any compound provided herein, or the drug thereof. This includes administering to the individual a composition (e.g., a pharmaceutical) containing a scientifically acceptable salt. , a method, for example, compounds are formulas I, II, III, IV, V, VI, V II, Formula VIII, Formula IX, Formula X, Formula XI, Formula XII, Formula XIII, Formula XIV, Formula XV, Any one of the following formulas: formula XVI, formula XVII, formula XVIII, formula XX, or formula XXX It has a structure. In some embodiments, the fibrous disease or disorder is fibrosis. In this embodiment, fibrosis is IPF, fatty liver disease, cirrhosis, colitis, chronic liver disease, cardiac fibrosis. The choice is made between vascular disease and scleroderma.

[0193] In certain embodiments, the herein provides a method for transferring folate to cells via a linker. Immunomodulatory substances attached to Gand or its functional fragments or analogs or their pharmaceutically A fibrous disease state comprising contact with at least one compound containing an acceptable salt A method of prevention or treatment, in which an immunomodulator or a pharmaceutically acceptable salt thereof is used. It targets the receptor that recognizes light.

[0194] In some embodiments of the compounds, compositions, and / or methods provided herein, The immunomodulatory substance includes a TLR agonist, which is expressed by the following formula X or XX. It is a structure that is either a pharmaceutically acceptable salt of formula X or XX. [ka] In the formulas, in formulas X and XX, R1 is either -NH2 or -NH-R 1X And, R2 is H, alkyl, alkenyl, alkynyl, alicyclic, aryl, biaryl, hete Royryl, -NH-R 2X , -OR 2X, -SR 2X , [ka] And, [ka] These are 3-10 member nitrogen-containing non-aromatic monocyclic or bicyclic heterocyclic rings. In equation X, R3 is -OH, -SH, -NH2, or -NH-R 1X And, In formula XX, X is CH, CR2, or N. R 1X , R 2X , and R 2Y Each of these can independently be H, alkyl, alkenyl, or alkini. Selected from the group consisting of aryl, alicyclic, aryl, biaryl, and heteroaryl compounds.

[0195] In some embodiments, cells are experiencing or will experience a fibrous disease state. The solution involves a sample containing target cells, and bringing these cells into contact with at least one compound. Further includes administering or applying an effective amount of at least one compound to the target. In that embodiment, the subjects are patients experiencing IPF, and at least one compound It is administered to the target intravenously, intramuscularly, intraperitoneally, topically, or by inhalation. In this embodiment, the fibrous disease state is IPF or the liver, skin, bladder, heart, pancreas, prostate This includes fibrotic diseases of the glands or kidneys.

[0196] In some embodiments, the method involves obtaining a sample from a subject, or obtaining a sample This involves quantifying the expression level of one or more biomarkers in the sample, Each of one or more biomarkers is CCL18, arginase 1 (Arg1), and Matri TX metallopeptidase 9 (MMP9), metalloproteinase 3 (TIMP3), I L-1β, hydroxyproline, collagen, PDGF, TGFβ, FRβ, TNFα, IFN-γ, anti-mannose receptor (CD206), differentiation antigen group 86 (CD86), differentiation anti Original group 163 (CD163), IL-6, chemokine 10 (CXCL10), immunotherapy Selected from the group consisting of ferron (IFNα), to be quantified, and one or more in the sample. The expression levels of each biomarker, compared to the expression levels of such biomarkers in the control. Compared with CCL18, Arg1, MMP9, TIMP3, IL-1β, PD GF, TGFβ, FRβ, CD206, CD163, hydroxyproline, or cholangiocarcinoma Is the gene upregulated compared to the control expression level, or is TN Fα, IFN-γ, IL-6, CXCL10, IFNα, or CD86 are expressed in the control group. If the level is downregulated or not present compared to the normal level, treatment Administer an effective amount of an unconjugated agonist or inhibitor to the subject. or administration of a folic acid ligand or The functional fragment or analogue of this substance is specific to FRβ and binds to FRβ on cells.

[0197] In at least one embodiment, fibrous diseases (e.g., IPF, etc.) are treated and / or Alternatively, methods for prevention are provided. The method involves M2-like macros in fibrous tissue or organs. To reprogram the phage to an M1-like phenotype, (by linker or other means) ) One or more compounds containing a targeting portion (such as a folate receptor-binding ligand) attached to the drug. This includes administering a therapeutically effective dose to the target. For example, the drug is a Toll-like receptor agonist. (For example, having formula I, III, or IV), or macrophages that contain folic acid Any other effective method for jugating and reprogramming from the M2 phenotype to the M1 phenotype It may be a molecule or a compound. In at least one embodiment, the drug is TLR 3 Agonist, TLR7 agonist, TLR7 / 8 agonist, TLR8 agonist, oyo A TLR9 agonist may be selected. In some embodiments, the drug is M2-like. Macrophages can be reprogrammed to the M1 phenotype, thereby promoting fibrosis. This can reduce the production of ions and growth factors.

[0198] In at least one embodiment, there is a risk of having or experiencing a disease state. A method is provided for treating the subject, where the disease state is a fibrotic disease state or The method includes contacting the target cells with at least one compound. At least one compound may include any of the compounds of this disclosure, and at least one example In an exemplary embodiment, the targeting moiety is specific to FRβ. In some cases, the cells are brought into contact. This involves administering at least one compound intravenously, intramuscularly, intraperitoneally, topically, or orally. Administered by or by inhalation, or by any of the other modes of administration described herein. This can be achieved by: Additionally or alternatively, at least one compound, The above pharmaceutically acceptable carriers, adjuvants, diluents, excipients, and / or vehicles The composition may include, or a combination thereof, at least one administered The dosage of the compound may be adjusted as appropriate by the clinician, but at least one compound , administered in a therapeutically or prophylactically effective amount, and in at least one embodiment, the dose is 1n The mol / kg concentration is within the range of 50 nmol / kg of the target body weight.

[0199] Referring now to Figure 2, one or more of the compounds of this disclosure can be used to treat fibrous disease symptoms. A representative flowchart of 1900 methods for preventing or treating a disease or cancer is shown. In at least one example, method 1900 uses a linker to transfer the target cells. , immunomodulatory substances attached to folic acid ligands or their functional fragments or analogs (or The step includes contacting at least one compound (containing a pharmaceutically acceptable salt thereof) with the product. (Step 1902). In at least one exemplary embodiment, an immunomodulator or so The pharmaceutically acceptable salt of targets pattern recognition receptors. Cells, for example, fibrous It may include target cells that are experiencing or at risk of experiencing a disease state, and a small number At least one compound may include any of the compounds provided herein.

[0200] In at least one embodiment, the step involves bringing cells into contact with at least one compound. 1902 refers to administering or applying a therapeutically effective dose of at least one compound to the target. It also includes. In addition or alternatively, at least one compound is pharmaceutically acceptable. Possible carriers, adjuvants, diluents, excipients, and / or vehicles, or their It may include compositions containing combinations.

[0201] In at least one embodiment, the disease condition is IPF of the liver, skin, heart, or kidneys. or including fibrous diseases. Furthermore, the subjects include mice, humans, or any other mammals. It may be included.

[0202] Method 1900 optionally includes steps 1904-1910 in addition to step 1902. That's fine. In step 1904, a biological sample is obtained from the subject, and in step 1906, the Quantify the expression level of one or more biomarkers in a sample. For example, the sample is from the target. It may be obtained from a certain amount of peripheral blood collected.

[0203] The quantitative step 1906 is performed using any suitable method known in the art. Common methods include, for example, qPCR, mass spectrometry, ELISA, and / or biomarker expression. It may include any other modalities that can measure / quantify at least one In an exemplary embodiment, one or more biomarkers are CCL18, Arg1, MMP9 TIMP 3, IL-1β, PDGF, TGFβ, FRβ, hydroxyproline, cholesterol -gen, TNFα, IFN-γ, CD206, CD163, IL-6, CXCL10, I Selected from the group consisting of FNα and CD86.

[0204] In step 1908, the expression levels of one or more biomarkers in the sample were compared to the control. The expression levels of such biomarkers are compared to those of healthy individuals. It could simply be an individual that has not experienced the disease condition in question. At least one implementation Morphologically, the expression levels of one or more biomarkers in the sample and the associated biomarkers in the control The clinical difference between the level of expression of Carr and the subject is that the subject is suffering from the disease state in question. It can be shown that, for example, but is not limited to, comparison step 1908 However, the biomarkers CCL18, Arg1, CD163, MMP9, TIMP3, IL- 1β, PDGF, TGFβ, FRβ, hydroxyproline, collagen, and / or The expression of one or more CD206 (i.e., "pro-fibrosis biomarkers") is in opposition to If it indicates that it is upregulated compared to the light, then this means that the subject is This demonstrates the experience of a pro-fibrotic immune response associated with the M2-like macrophage phenotype. Therefore, in at least one embodiment, such results are like M2. To reprogram macrophages to the M1 phenotype, one or more compounds of this disclosure are used. This indicates the necessity of administering the medication.

[0205] In contrast, comparison step 1908 showed that the expression of the aforementioned biomarkers was compared to the control. If it indicates that it is downregulated, or if one or more TNFα Expression of IFN-γ and / or CD86 ("anti-fibrotic biomarker") is controlled by the control group. If it is upregulated compared to, this is in a particular embodiment, versus Whether the elephant shows a positive response to a previously administered compound (if applicable), and / or This indicates either that the subject is experiencing an anti-fibrotic immune response associated with the M1 phenotype. .

[0206] Optionally, in step 1910, the development of one or more fibrosis-promoting biomarkers in the sample. If the current expression level is upregulated compared to the control level, Alternatively, the expression of one or more anti-fibrotic biomarkers is compared to the respective expression levels in the control group. In contrast, if downregulation occurs in the sample, alternative therapies can be administered. In at least one embodiment, the alternative therapy is administered previously in step 1902. This may include administering a therapeutically effective amount of a derivative of at least one compound, the derivative This involves different targeting regions, different linker sizes, and / or different immunomodulators. Regarding adopting either approach to better optimize the effectiveness of at least one compound Contains at least one modified, previously administered compound. Additionally or alternatively, Other treatments, including those conventionally known for treating the fibrous disease in question. This may be adopted. Steps 1904-1910 may be performed as needed or as desired. It can be repeated.

[0207] An additional embodiment involves treating cancer (whether folate receptor-positive or folate receptor-negative). It may provide methods for treatment and / or prevention. For example, in certain cases, such The method involves reprogramming M2-like macrophages in cancerous and / or tumor cells to an M1-like phenotype. Includes a targeting portion attached to the drug (by a linker or other means) for programming. Administering a therapeutically effective and / or prophylactically effective dose of one or more compounds to a host subject. This includes. If the cancer is folate receptor-negative, such administration is in such tissue / tumor. It may further act to deplete or inhibit the MDSCs present in [the location]. For example, [the following may be limited to] However, the drugs include PI3k inhibitors and signaling and transcriptional activator 6 (STAT6) inhibitors. Harmful agents, mitogen-activated protein kinase (MAPK) inhibitors, inducible nitric oxide compounds Selected from enzyme (iNOS) inhibitors and anti-inflammatory drugs (e.g., methotrexate) It is possible. In at least one embodiment, the drug can inactivate MDSCs.

[0208] In describing typical embodiments, this disclosure describes the method and as a specific set of steps. / or may present a process. The method or process is specified herein. Unless the steps depend on a specific order, the method or process is the same as the steps described. It should not be limited to a specific order. As those skilled in the art will understand, the steps Other orders may be possible. Therefore, the specific order of the steps disclosed herein is Furthermore, the method and / or patent The claims directed at the process are limited to the execution of those steps in the order described. It should not be fixed, and a person skilled in the art may change the order, and the essence of this disclosure remains the same. It is easy to understand that this is God and within the realm of God. [Examples]

[0209] Human monocytic THP-1 cells, American Type Culture Col Obtained from the collection, 10% heat-inactivated fetal bovine serum and 1% penicillin / stress Folic acid deficiency R containing ptomycin (Invitrogen, Carlsbad, CA) The cells were cultured in PMI1640 medium (Invitrogen, Carlsbad, CA). HP-1 cells are a human monocyte cell line that has acquired an M2-like phenotype and IL-4, IL-6, and It is known that stimulation with IL-13 produces a significant amount of pro-fibrotic cytokines. Because of this, she was initially selected as a model.

[0210] IFN-γ, IL-4, interleukin-6 (IL-6), and interleukin -13 (IL-13) was obtained from Biolegend. Holball 12-Millista 13-acetate (PMA), lipopolysaccharide (LPS), and all other reagents and solvents. I bought it from Sigma.

[0211] Example 1: In vitro differentiation and polarization of THP1 cells into M2-like macrophages THP-1 cells were seeded into 96-well plates at a density of 60,000 cells / well. The cells were incubated with 200 nM PMA for 48 hours, followed by fresh RPMI. By incubating in culture medium for 24 hours, the cells are differentiated into nonpolar macrophages. The obtained macrophages were treated with 20 ng / ml IL-4 and 20 ng / ml IL-1. M2-like cells were incubated with 3 and 5 ng / mL of IL-6 for 3 days. The phenotype was polarized, and then reprogrammed for 48 hours with different concentrations of compound 1A and compound 1B. Gram-sized samples were collected for genetic analysis by quantitative polymerase chain reaction (qPCR). The cultures were maintained at 37°C in a humidified 5% CO2 incubator.

[0212] A potent TLR7 agonist (e.g., compound 1A of formula III) promotes fibrosis. Whether advanced macrophages can be reprogrammed into a less fibrillating phenotype. To evaluate, THP-1 cells stimulated with IL-4, IL-6, and IL-13 were used. This was incubated with non-target compound 1A at different concentrations, and several fibrosis-promoting markers were identified. That is, the mR of CCL18, CD206, IL-1β, and PDGFα and β I checked the NA level.

[0213] As shown in Figures 3A-3C, the 48-hour incubation with compound 1A was CCL1 8. TLR7 agonists induce a decrease in CD206 and IL-1β expression, and these The shift of THP-1 cells polarized to promote fibrosis to a lower fibrous phenotype is actually This suggests that it can be promoted. Furthermore, the expression of TNFα, a marker of the anti-fibrotic phenotype, Upon investigation, an increase in its expression was observed (Figure 3D), and THP-1 is thought to have fibrosis-promoting properties. It was confirmed that the properties shifted to promote fibrosis.

[0214] Example 2: Evaluation of macrophage reprogramming A TLR7 agonist conjugated to folic acid is the same THP found in Example 1. -1 To confirm that it can induce reprogramming, compound 1B was prepared. This enables the internalization of compound 1B into the reducing environment of intracellular endosomes, followed by the release of compound 1A. To enable this, a releaseable linker is attached to compound 1A with folic acid, and a disulfide is used. It was constructed using a combination of methods.

[0215] Either compound 1A or compound 1B at different concentrations is applied to the above polarized THP-1 macromolecule. Lophages were incubated for a specified time, followed by analysis of secreted cytokines and qPC. Culture medium was collected for cell collection for R analysis.

[0216] Follow the manufacturer's recommended protocol to use Quick-RNA® MicroPre Using the p kit (Zymo Research, Irvine, CA), 1 × 10 5 ~2×10 5 Total RNA was isolated from macrophages. Then, the RNA sample was subjected to a large volume of water. Quantitative cDNA reverse transcription kit (Applied Biosystems, Foster Ci) The result was reverse transcribed into cDNA using ty,CA, #4368814). qPCR analysis was performed using i Taq (trademark) Universal SYBR Green SuperMix (Bio -Rad Laboratories GmbH, Hercules, CA, #1725 121), iCycler Thermocycler, and iCycler iQ 3.0 Soft Toware (Bio-Rad Laboratories GmbH, Hercules, Using CA), we tracked the expression of markers characteristic of macrophage polarization states. . As markers for the M1 phenotype, IL-6, CXCL10, IFNα, IFN-γ, and Using CD86, CCL18, CD206, and CD163 were used as markers for the M2 phenotype. , and Arg1 were adopted. IL-1β, PDGFβ, MMP9, and TIMP 3 This was measured as an indicator of the fibrotic phenotype. IRAK-4 was used as an indicator of TLR7 stimulation. The following was used. To control the specificity of the amplified product, melting curve analysis was performed. However, no amplification of nonspecific products was observed. Each sample was 3 for each marker. Each street was analyzed independently.

[0217] Repeating the above research (see Figures 3A-3F, gray bars) reveals the same qualitative changes A change was observed, and only the magnitude of the effect of compound 1B was slightly reduced. Non-targeted TLR7 While gonists immediately enter cultured cells, their folate-targeted counterparts enter folate-receiving cells. Designed to enter cells only after receptor binding and receptor-mediated endocytosis. Therefore, a decrease in effectiveness was anticipated.

[0218] Figures 4A-4E and 5A-5D show THP-1 cells induced into M2 macrophages. The graph shows data representing various marker levels measured from, and these are then shown as different... Incubate with compound 1B or compound 1A at the specified concentration for 2 hours, wash with PBS, and see Figure 5. The data shown in A-5D were incubated again for 46 hours (Figures 4A-4E). For the data shown, the cells were collected immediately after the first two hours of incubation. (This was done.) Cells were collected for genetic analysis by qPCR in both datasets. Figures 4A-4C show CCL18 mRNA levels (Figures 4A and 5A), CD20 6 mRNA levels (Figures 4B and 5B), IL-1β mRNA levels (Figure 4C and 5B) Figure 5C) and PDGFβ mRNA levels (Figure 4E) are shown. The data are M2-like lines. The fact that the fibrosis-promoting phenotype was downregulated after administration of the tested compound supports the claim that It is attached. In particular, compound 1B promotes macrophage fibrillation / M2 type marker, It was downregulated more significantly than compound 1A. Furthermore, Figure 4D shows CD86 mR Figure 5D shows NA levels and TNFα levels, and these data indicate an M1-like phenotype. However, this supports the idea that upregulation occurred after administration of the tested compound. During the treatment period, no significant difference was observed after treatment, therefore, the data regarding PDGFα were not presented. It hasn't been done.

[0219] Low molecular weight water-soluble drugs such as compound 1A and compound 1B are often administered by injection. Because it is excreted from the body within a few hours, it is a more physiologically appropriate method for in vivo drug exposure. The in vitro model limits the incubation of cells with drugs to only 2 hours, and the drug's effect The efficacy of the drug will be examined after a further 46-hour incubation period. (Figures 4A-4E) As shown, after incubating THP-1 cells with a TLR7 agonist for 2 hours, the drug When the drug-containing culture medium is replaced with a drug-free culture medium, compound 1B exhibits superior performance compared to compound 1A. It has been observed that it has a specific effect, and in particular, in the case of TNFα induction, folate-targeting conjugates The condition dramatically improved. This is because folate-targeted TLR7 agonists act on folate receptor-positive cells. This is because compound 1A was captured, whereas compound 1A was not retained by the same cells. It seems so.

[0220] These data show that compound 1B reprograms pro-fibrotic macrophages in vivo. Folic acid-conjugated drugs should be more effective in molting. (For example, compound 1B) is concentrated in FRβ-expressing macrophages and is dominant throughout the body. Acid receptor-negative cells (for example, compound 1B is impermeable to folate receptor-negative cells) (It is designed to prevent) entry, and therefore does not cause more systemic toxicity. This supports the idea that it has further advantages.

[0221] Figures 6A-6D show treatment with different concentrations of the drug for 48 hours (Figures 6A and 6B) or 2 hours. Afterward, the medium was replaced with fresh medium and incubated for the remaining 46 hours (Figures 6C and 6D) M2 Graph data representing various marker levels measured from induced THP-1 macrophages. In both cases, the cell supernatant is collected, and the secreted CCL18 protein and IL are shown. -1β was detected by ELISA. The data targeted TLR7 compounds or folic acid. Administration of TLR7 compounds at low concentrations (0.1-10 nM) affects CCL18 and IL-1 This supports the idea that it downregulates β secretion.

[0222] Furthermore, the above mRNA analysis was found to be produced by IL-4, IL-6, plus IL-13. To accurately reflect the levels of fibrosis-promoting cytokines, stimulate THP-1 cells. Therefore, the concentrations of CCL18 and IL-1β polypeptide in the THP-1 supernatant are controlled by ELI Quantitative analysis was performed by SA assay. As shown in Figures 6A and 6B, compound 1A and chemical When both compounds 1B were incubated with the agonist for 48 hours, CCL1 It induced a decrease in 8 and IL-1β, but the compound was only activated when drug exposure was limited to 2 hours. Item 1B was again found to be superior (see Figures 6C and 6D).

[0223] Example 3: Characterization of FRβ expression by flow cytometry To measure FRβ expression in THP-1-derived macrophages, fluorescence-activated cell selection was performed. A separate (FACS) analysis was performed. The cells were analyzed using Accutase ( (Registered Trademark) Cell Exfoliation Solution (Use Biolegend, San Diego, CA, #423201) to remove the stripping agent. The cells were gently lifted with a cell scraper. The cells were washed with PBS and Fc receptor blocks were removed. Using a sizing solution (Biolegend, San Diego, CA, #422301) Nonspecific binding was blocked by incubation at room temperature for 10 minutes. Next Then, biotinylated anti-human FRβ monoclonal antibody (m909) was added, and the cells were placed on ice. After incubation for 30 minutes, in staining buffer (PBS supplemented with 2% FBS) The cells were washed. Then, the cells were treated with fluorescein-labeled streptavidin (BD Biosci). (Franklin Lakes, NJ, #554060) spent 20 minutes on the ice. Incubate in PBS, wash twice, stain with 7AAD (viability stain) for 15 minutes, BD Accuri C6 software (BD Biosciences, Frankl The analysis was performed using flow cytometry with Lakes (NJ). Figure 6E shows: The flow cytometry data shows that the THP-1 macrophage is FRβ+, and Therefore, it is suitable for in vitro studies of compound 1B and other studies described herein. This supports the idea.

[0224] Figure 6F shows that compound 1B remained stable during the incubation period at 37°C in the culture medium. Confirm that it remained unchanged. In fact, compound 1B after 48 hours of incubation It retained its original structure.

[0225] Example 4: In vivo study of bleomycin-induced pulmonary fibrosis and pro-fibrotic macrophages Reprogramming Can macrophages in pulmonary fibrosis be specifically targeted in vivo with folate-conjugated agents? Tests were also conducted to determine whether or not this was the case. Multiple studies were conducted on inducing pulmonary fibrosis in mice. After testing the protocol, 0.75 mg / kg of bleomycin (BM) was administered via tracheostomy. It was injected into the lungs of C57BL / 6 mice via [method], and the inflammatory stage of fibrosis and fibrosis were observed before the start of treatment. We selected a protocol that proceeds to mice through both stages of development. (The BM model is an Inv It is considered to be widely useful in that it enables mechanistic investigations related to fiber formation in the context of the Bo situation. (It is being done.)

[0226] As shown in Figures 7A-7D, mice treated using this protocol are typically Fibrosis was presented by the 7th day after BM treatment, and this neofibrosis progressed to severe fibrosis by the 14th day. It develops into a vascular disease. The progression of the disease then continues for 2 to 5 days and resolves spontaneously by the 21st day. start.

[0227] More specifically, 8-week-old C57BL6 male mice from Charles River ( (Average body weight 22g-25g) Under pathogen-free conditions, at room temperature (22℃) for 12 hours in light-dark conditions. The mice were reared under irrigation conditions. The mice were fed a folic acid-deficient diet (En) for one week prior to BM or PBS injection. I chose Vigo Teklad Global Rat Food Pellets. Fresh water and a folate-deficient diet were freely available. All animal procedures were conducted by the National Institutes of Health. In accordance with the guidelines, Purdue Animal Care and Use Co. Approved by mmittee

[0228] Afterward, the mice were anesthetized with ketamine / xylazine, and the necks of the mice were treated with hair removal lotion. The area was shaved using a razor, then sterilized with 70% alcohol. A small incision was made in the neck to visualize the trachea. The procedure was performed. The mouse was positioned at a 75-degree angle, and a 1cc syringe with a 26G needle was used. 100uL of sterile PBS or BM (Cayman Chemic) dissolved in PBS als (Ann Arbor, MI, #13877) (0.75 mg / kg) administered intratracheally. An injection was administered. Throughout the experiment, body weight was monitored every other day.

[0229] These mice exhibit pro-fibrotic lung macrophages that are specifically targeted by folate-conjugated agents. To evaluate whether it can be converted, 10 nmol (for in vivo imaging) was administered 10 days after the injection. This is 200 times the excess of 100 nmol (for in vivo labeling) of FA-glucosamine (OTL38) Folic acid-linked near-infrared fluorescent dye (OTL38), which may or may not contain (competitors), is treated with BM. The dye was injected into the tail vein of mice, and its uptake into major organs was evaluated.

[0230] Two hours later, the mouse was sacrificed using CO2 asphyxiation, and the skin from the abdomen to the neck was immediately cut open. Then the lungs and trachea were exposed. Next, a 22-gauge blunt needle was inserted into the upper trachea. Make a small cut and tie a nylon string around the trachea to seal the area around the needle. The opening was closed. Then, the trachea (including the insertion needle), lungs, and heart were carefully cut through the connective tissue beneath the lungs. By cutting and consolidating, it is removed, and the bronchi of the left lung are defused. The Bach vascular clip was used to clip the lung. PBS was injected into the right lung using a 1 ml syringe. The solution was aspirated three times, and the collected cleaning solution was stored on ice.

[0231] Next, bronchoalveolar lavage fluid (BALF) was analyzed to determine which targeted TLR7 agonists were effective. We determined how it works. The BALF sample was centrifuged at 1500 rpm for 5 minutes at 4°C. The supernatant was aliquoted and stored at -80°C for cytokine / chemokine analysis. Resuspend the cell pellet and culture it in pre-warmed RPMI1640 medium for 2 hours, then... The samples were then washed three times with pre-warmed PBS before collection for the qPCR assay. The lungs were bound together with nylon strings and used for subsequent analysis of hydroxyproline content. The left lung was inflated with 1 ml of PBS using an inserted syringe, followed by histological analysis. It was then transferred to a 10% formalin solution.

[0232] The right lung lobe collected above was weighed and placed in a pressure-resistant vial (Supelco Inc., Bel Place in lefonte, PA, #27003) and bathe in sand at 120°C for 3.5 hours, 6N The solution was hydrolyzed with HCl (10 ml / g, v / w). The hydrolyzed solution was incubated at 4°C for 15 minutes. After cooling and transferring to a 1.5 ml Eppendorf tube, incubate at 12,000 rcf at 4°C. The mixture was centrifuged for 15 minutes. The supernatant was carefully collected and separated, and hydroxyproline (HYP) was added. ) Used in the analysis.

[0233] In the subsequent HYP analysis, 10 μl of the sample was transferred to a 96-well plate, and 10 μl of 5 It was neutralized with 0.3M sodium hydroxide solution. Then, isopropanol (40 μl) was added to each of the following containers. Add to the gel, then add 20 μl of oxidative buffer, and shake the mixture at room temperature for 5 minutes. The plate was incubated above. Analysis reagent (260 μl) was added, and the plate was incubated at room temperature for 30 seconds. The samples were incubated on a shaker and immediately incubated at 60°C for 25 minutes. Absorbance Within 15 minutes, 560nm (A 560 ) was measured. All reagents were previously reported. It was prepared according to the protocol.

[0234] For histological analysis of lung sections, the fixed lung (see above) is embedded in paraffin. Section the material and treat with hematoxylin-eosin (H&E), Masson's trichrome or F3( The tissue sections were stained with anti-mouse FRβ antibody. The tissue sections were then examined in a blinded manner by an authorized pathologist. I looked it up. Aperio-Image Scope (Leica Biosystems, Using Wetzlar (DE), 90 × 10 for each section. 6 Quantitative functions for more than one cell Ta.

[0235] Human DuoSet ELISA Development System (R&D Systems Europe) (Abingdon, UK, #DY394-05) and IL-1 beta human ELISA Kit (Thermo Fisher Scientific, Waltham, MA; #BMS224-2) is used as described by the manufacturer, and the induction CCL18 and IL-1β were quantified in the THP-1 cell supernatant. BALF samples were then processed using E LISA MAX(TM) Deluxe (Biolegend, San Diego,C Mouse IFN-γ was analyzed using A;#430804).

[0236] Finally, regarding the in vivo folate imaging study, the major organs (heart, lungs, spleen, liver, small intestine, The large intestine and kidneys were removed, and an AMI live imager (Spectral Inst) was used. Images were acquired using Brüns Imaging (Tucson, AZ). In vivo For the folate receptor labeling test, the lungs of mice are collected immediately after euthanasia and explained in the manual. As shown, gentleMACS Octo Dissociator with Heathers(Miltenyi Biotec, Bergisch Gladb Following the instructions for ach,DE, #130-096-427), use the lung dissection kit (Milte nyi Biotec, Bergisch Gladbach, DE, #130-098 Digestion is performed using -427) and a 70μm cell strainer (Miltenyi Biote c,Bergisch Gladbach,DE, #130-098-462) filtration The cells collected in the filtrate were depleted of red blood cells by ammonium sulfate dissolution, and then cooled with PB. Wash twice in S, then use an antibody against the desired macrophage marker and leave on ice for 30 minutes. Labeled (FITC-CD11b, Biolegend, San Diego, CA, # 101205;FE-F4 / 80, Biolegend, San Diego, CA, # 123109). Next, the labeled macrophages are washed twice in PBS and 7AAD Stain with (viability staining) for 15 minutes, then use BD Accuri C6 software (BD Bi Analysis using flow cytometry with osciences (San Jose, CA). did.

[0237] As shown in Figure 7A (upper panel), untreated lung (PBS control column) and B on day 7 M-treated lungs exhibit similar high-density alveoli interconnected by minimal extracellular matrix. In contrast, on day 14 after BM injection, the size and frequency of air sacs decreased significantly, and the extracellular matrix was also reduced. The density of the cells was clearly increased, suggesting the development of significant fibrosis in the treated mice. By day 21, the symptoms in this model had already begun to resolve spontaneously, and many mice were the most Ultimately, the patient recovered from the BM-induced trauma by day 35.

[0238] Evidence of inflammation development up to day 7 was found in the infiltration of FRβ-expressing macrophages (Figure 7). (See lower panel A and quantification in Figure 7B), it is almost completely present in healthy lungs. However, in lungs exposed to BM, it continues to accumulate until day 14. Furthermore, staining with F3 The color indicates significant FRβ in IPF lung (mainly interstitial space), as previously reported in the literature. Expression was observed (Figure 7A). FRβ expression was limited to the inflamed lungs (IPF patients). (either BM-induced PF, but not observed in healthy lungs). Furthermore, F Rβ-expressing macrophages were observed in mouse lungs on day 7 after BM administration, reaching their peak on day 14. This was the expression (Figure 7B). These results are consistent with previously reported activity in inflamed lungs. This confirmed FRβ expression in sexualized macrophages.

[0239] Next, we can demonstrate that these FRβ-expressing macrophages can be targeted by folate-binding molecules. Folic acid-targeted fluorescent dye in the lungs of mice that were BM-treated but unhealthy after tail vein injection. This was demonstrated by the accumulation of OTL38. As shown in Figure 7B, OTL38 fluorescence is healthy It was observed only in the kidneys of healthy mice (i.e., their primary excretory site), and not in other tissues. There was little to no congestion.

[0240] Figures 7C and 7D show human IPF lung tissue (Figure 7C) and healthy human lung tissue (Figure 7D). This shows FRβ IHC staining. 8-week-old C57BL / 6 male mice were BM or PB. For one week prior to the injection of S, the mice were fed a folic acid-deficient diet, and 10 days after the injection, 10 nmol (I) was given to the mice. FA-gluco (for in vivo imaging) or 100 nmol (for in vivo labeling) in 200-fold excess OTL38, with or without samine, was injected via the tail vein. Two hours later, Mau The spleen was sacrificed before analysis. For in vivo folate imaging studies, major organs (heart, lungs, spleen) were targeted. The organs (liver, small intestine, large intestine, and kidneys) are removed, and an AMI live imager (Spect Taken using ral Instruments Imaging (Tucson, AZ). The image was taken. For the in vivo folate receptor labeling test, the lungs of mice were collected immediately after euthanasia. Digestion, followed by the desired macrophage markers (FITC-CD11b, PE-F4 / 8) Labeled with antibodies against 0) and 7AAD (live / dead staining), and then subjected to flow cytometry. I analyzed it.

[0241] Figure 7E shows mice with or without BM-induced experimental fibrosis (BM) (PBS control). Various mouse tissues were collected and imaged with the folate receptor-targeting fluorescent dye, OTL38. Images of organs are shown, comparing healthy mice (column a) and BM-treated mice (columns b and c). Ten days after inducing fibrosis in the tail vein, 10 nmol of OTL38 was detected, resulting in a 200-fold excess of folate target. (b) Absence of glucosamine (a competing reagent for FRβ that blocks OTL38 binding) The patient was injected with (c) and euthanized two hours later for tissue excision and fluorescence imaging. This is because the FA-targeted conjugate of the invention disclosed herein is absorbed by other healthy tissues. This supports the idea that it exhibits FRβ-specific binding without any complications.

[0242] Injection of OTL38 into the tail vein of BM-treated mice resulted in the aforementioned fluorescence in the kidneys, It also resulted in significant accumulation in fibrotic lung (see Figure 7E). This uptake into the lung is primarily It was found that folate receptors mediated the process, and BM-treated mice received a 200-fold excess of folate-glucoglycan. When samin (i.e., a competitive inhibitor of FRβ binding (see Figure 7E)) is injected simultaneously) This can be demonstrated by nearly quantitative blockade of lung accumulation. These data suggest that folate target molecules are... Without significant accumulation in other tissues of the body, folate receptors develop in fibrous tissues. This indicates that it selectively binds to existing cells. In other words, FRβ-expressing macrophages are actually In such cases, it can be targeted with folic acid-binding molecules, and in clinical applications, it is localized almost exclusively to fibrous tissue. It is present. Therefore, when using the target moiety in the compounds of this disclosure, the fibrous nature of the target ( Any TLR7 agonists not captured by (or cancerous) tissue are minimal.

[0243] Next, we will examine which cell type captures the folate-dye conjugate in the lungs of BM-treated mice. To determine if it is a dolphin, the lungs of the above animals were digested with collagenase and flow cytometry was performed. We investigated cell-specific pigment uptake using this method. Figure 7F shows the results of BM-induced experimental fibrosis. This shows data from FACS analysis obtained from in vivo labeling of the mice being tested. This involves targeting folate in PBS (column 1) or 100 nmol OTL38 in 200-fold excess. In the absence (column 2) or presence (column 3) of glucosamine, when injected into the tail vein. Yes, as shown in Figure 7F, OTL38 was isolated from BM-treated mice that had not been injected. In this case, macrophage-like cells did not exhibit fluorescence (see column 1). In contrast, OL Approximately 22% of macrophage-like cells derived from fibrous mice injected with T38 showed significant folate targeting It shows retention of color pigment (column 2), which is FRβ-positive macro in the inflamed lungs of OTL38. This supports the idea that it targets phages. In fact, 200 times the amount of folate-glucosamine The observation that simultaneous tail vein injection essentially blocked all folate-pigment retention proved true. As evidenced, pigment uptake is particularly folate receptor-mediated, and pigment accumulation is occupying This demonstrates the need for folate receptors that are not present. Importantly, this conclusion is that FRβ expression is essentially undetectable in untreated lungs (see Figure 7A), but in BM-treated lungs... Data showing a dramatic increase during the development of fibrosis in the lungs (see Figures 7A-7D) This is further supported by the fact that FRβ expression is also significantly present in the lungs of human IPF patients. It is showing.

[0244] Example 5 Next, the ability to target drugs attached to established FRβ-expressing fibrillary macrophages. Using this method, folate-targeted TLR7 agonists were found to reduce signs of fibrosis in BM-treated mice. The question was whether the symptoms could be suppressed. For this purpose, BM-treated mice were given Use either the vehicle (3% DMSO in PBS) or compound 1B every other day starting from day 10. It was administered by intravenous injection (see Figure 8A). The TLR7-54 agonist caused rapid weight loss and Compound 1A was also used in vivo because it caused subsequent death (see Figures 9A and 9B). It could not be evaluated. In BM-induced experimental pulmonary fibrosis in mice, inflammation occurred approximately after BM induction. It is known to last for 9-10 days. Therefore, the transition from inflammation to fibrosis is In this model, this occurs approximately between day 9 and day 14, and fibrosis-promoting markers appear around day 10. The symptoms began to subside, and medication was started on the 10th day (Figure 8A).

[0245] The drug was administered twice, every other day, until day 21. This prevents any "resistance" to the TLR agonist. Therefore, the individual daily doses are divided every 6 hours. Next, on the 21st day, the mice are sacrificed. The lungs were immediately subjected to bronchoalveolar lavage, followed by immunohistochemistry and analysis of collagen and hydroxylase. A lung was resected to quantify cyproline levels.

[0246] Figures 8B-8G show various markers measured from mice treated with the BM model in Figure 8A. - The graph shows the level, where BALF was collected on day 21 and centrifuged at 4°C. Separate the pellets, resuspend them in culture medium, seed them in a 96-well plate, and incubate for 2 hours. Then, the cells were washed three times with pre-warmed PBS, and the cells were collected for qPCR. The tools used are Arg1 (Figure 8B), MMP9 (Figure 8C), and TIMP 3 (Figure 8D) (for example, lines). This indicates that all of the fibrosis-promoting markers (CD86) are downregulated. (Figure 8E) ) and IFN-γ (Figure 8F) (e.g., an anti-fibrotic marker) are both upregulated Furthermore, the number of BALF cells present (Figure 8H) was also analyzed, as was the TLR7 signature. The negative regulator of the nal transmission IRAK-4 was upregulated (Figure 8G). Furthermore, the total number of mouse BALF cells was dose-dependent after treatment with different doses of compound 1B. It decreased to [value]. The values ​​shown in Figures 8B to 8G represent the mean ± SD for each group. *P<0.05, **P<0.005, ***<0.0005; physiological saline vs. vehicle group For the treatment groups of compound 1A and compound 1B versus the vehicle group, Student's t-test was performed. Calculated by Dunnett's multiple comparison method, where compound 1B was used for both the treatment group and the vehicle group. Excluding BALF cell count and protein concentration measurements calculated by the test. Vehicle = 3% DMSO in PBS.

[0247] As shown in Figures 8B-8D, the lines in the macrophage subgroup of bronchoalveolar lavage cells qPCR analysis of vascularization-promoting markers revealed that BM-induced mice had a different odor compared to control mice. Furthermore, it was found that the tissue inhibitors Arg1, MMP9, and TIMP3 were all elevated. It became smoother. More importantly, in a parallel study, BM-induced mice were treated with compound 1B. When this happens, all the same fibrosis-promoting markers are suppressed, similar to those seen in healthy mice. It was demonstrated that levels of fibrosis-promoting markers were obtained. These data are consistent. The quantification of fibrosis-promoting markers was performed using CD86 transcripts (qPCR) and IFN-γ. It was revealed that both concentrations (ELISA of the washing solution) increased after treatment with compound 1B. (See Figures 7E and 7F). Observed IRAK-4 (i.e., T Markers of LR activation, upregulation of (results shown in Figure 7G), and The total number of BALF cells present decreased in a dose-dependent manner after treatment with different doses of compound 1B. In addition to the fact that (Figure 7H) these data indicate that folate-targeted TLR7 agonists Administration of this substance promotes fibrosis in the lungs of BM-treated mice in vivo. This demonstrates that it is possible to reprogram the phenotype from a fibrotic M1-like phenotype to a pro-fibrotic M1-like phenotype. .

[0248] Example 6 The aforementioned reprogramming of fibrous lung macrophages leads to a fibrous state in fibrous mice. Additional research was conducted to determine whether it resulted in actual improvement. Lung tissue from mice was embedded in paraffin, sectioned, and then treated with H&E and Masson's Triclove. The tissues were stained with M, and tissue density and extracellular collagen deposition were evaluated.

[0249] Figures 9A and 9B show experimental lung cells treated with untargeted and targeted TLR7 agonists. The survival curve (Figure 9A) and body weight change (Figure 9B) of mice with vascular disease are shown. The data are as follows: Administration of the compounds of this disclosure (here, for example, compound 1B) causes significant weight loss. This supports the idea that BM treatment increases the survival of mice without any adverse effects. Each value represents the average of each group. This represents the mean ± SD.

[0250] Figure 10A shows hydroxyprolyx in lung tissue, which uses collagen deposition as a measure of fibrosis. The content (μg / lung) is shown. The following tissues on day 21 are shown for each: healthy control (living (●) Saline solution, disease control (vehicle), free drug TLR7 agonist (compound) Treatment with 1A) (▼), treatment with a folic acid-targeted TLR7 agonist (compound 1B) (▲ ). B treated with 10 nmol of either compound 1B(▲) or compound 1A(▼). M-induced mice showed a higher total hydroxyproline content per lung compared to vehicle controls (■). This showed a significant decrease. Each value shown in Figure 9A represents the mean ± SD for each group. *P<0.05, **P<0.005, ***<0.0005, Student's t-test The results were as follows: physiological saline versus vehicle group, and compound 1A and compound 1B treatment group versus vehicle group.

[0251] Figures 10B and 10C show the results of H&E staining (Figure 10B) and Masson's trichrome (C). Figure 10A shows stained images of lung tissue using lagen staining (Figure 10C).

[0252] As shown in the H&E staining of the panel in Figure 10B, healthy lungs are surrounded by a thin reticular membrane. It is rich in air sacs. In contrast, BM-induced lung is characterized by extracellular matrix where air sacs once existed. It shows far fewer alveoli with significant deposition of quality. Most importantly, compound 1B is 10 Mice treated with BM injection from day 1 showed lung structures similar to those of healthy mice. Figure 10B) shows that fibrous lung macrophages suppress the major and prominent features of pulmonary fibrosis. This suggests that targeting compound 1A is effective. This prevention of fibrosis is actually Blocking of collagen deposition is associated with the Massontric morphology of parallel lunch sections. Recorded by loam staining (Figure 10C), collagen staining was performed by injecting compound 1B into the tail vein. It is strongly suppressed in the injected mice (Figure 10B). Therefore, the data is at least IPF mice treated with compound 1B(▲) showed suppression of IPF symptoms (e.g., fibrosis). This supports the idea that...

[0253] Finally, we confirmed that compound 1B actually influenced collagen production in vivo. To do this, hydroxyproline (a major component of collagen) in the total hydrolysate of affected lung tissue The components were quantified. More specifically, lung tissue from the above mice was perfused with PBS and then treated with acid. Hydrolysis was performed and the hydroxyproline content was analyzed. In Figure 10A, the induction of fibrosis was... It induced a significant increase in droxyproline content, and this increase was suppressed by treatment with compound 1B. Therefore, this data is relevant to the therapeutic use of the targeted TLR7 agonist compounds of this disclosure. The treatment reduces collagen deposition and therefore fibrosis in vivo (and furthermore, To support (opposing).

[0254] In summary, the overall survival rate of mice injected with the optimized BM dose (0.75 mg / kg) was reduced. The rate was significantly improved with treatment with compound 1B, but compound 1A resulted in significant weight loss. Except for showing a small improvement (>25%, Figure 7), there was no benefit in survival rate. The free drug was hydroxy It performed well in reducing cyproline content, but the observed poor survival rate was overall Possible cause: toxicity (i.e., weight loss; see Figure 7B). TLR7 This is surprising, given that systemic administration of agonists is known to cause toxicity. That wasn't the case.

[0255] Example 7 Use of non-targeted TLR7 agonists to treat IPF (or other fibrotic diseases) However, this is prevented by systemic activation of the immune system and the resulting toxicity, Evaluate whether compound 1B has apparent toxicity in Uss, potentially involving systemic administration. For this purpose, BM-induced mice were subjected to 0, 1, 3, or every other day starting from day 10. Treatment with 10 nmol of compound 1B, followed by body weight, lung hydroxyproline content, and histological analysis. It was performed on day 21. Unlike conventional systemic administration, the targeted drug not only improves survival rates but also Furthermore, weight loss, which highlights the significance of the targeted approach, was also reduced (Figures 11A and 11B).

[0256] Figure 12 shows lines in BM-induced mice, using collagen deposition as a measure of fibrosis. This paper presents data on the dose-dependent effect of folate-targeted TLR7 agonists on the suppression of vascular disease. The data is represented as follows: healthy control (PBS, ●), vehicle (■), 1 nmol compound 1B (○), 3 nmol compound 1B (□), or 10 nmol compound 1 BM-induced mice treated with B(▲), subpart A shows the time course of BM induction in mice. The graph shows weight-related data, and subpart B shows different doses (compound 1B at 10nm). Hydroxyproline content of lung tissue treated with ol, 3 nmol, or 1 nmol The measurements are shown, and subpart C shows a set of right lung tissue samples using H&E staining and trichrome staining. Images for textile analysis are shown.

[0257] Figures 11B and 12 subpart A show compound 1B in amounts of 0, 1, 3, or 10 nmol. No difference in weight loss was observed among the mice treated with the compound, which suggests that repeated administration of the compound may not be effective. Therefore, this suggests that no serious toxicity occurred. These measures still However, the expected effect against pulmonary fibrosis is still due to various pulmonary hydrolysates. This can be seen from a comparison of droxyproline content, and the order of effectiveness is 10 nmol / mau The order was S > 3 nmol / mouse > 1 nmol / mouse > 0 nmol / mouse (Figure 12). (See subparts B and C). More importantly, a detailed analysis of the lung tissue features is required. This shows that the lung tissue image improved as the dose of compound 1B increased, which is TLR The tissue with the strongest agonist concentration was the tissue that actually had the most normal microscopic morphology. This suggests something. In summary, these data indicate that TLR7 in fibrous tissue Targeting FRβ+ macrophages with TLR7 agonists in humans This supports the idea that fibrosis can be effectively prevented without systemic activation of the immune system that restricts it. It is.

[0258] Finally, in order to determine whether this antifibrotic effect can be achieved at low doses, two low doses were used. Treatment studies were conducted using doses (3 nmol / kg and 1 nmol / kg) (Figure 9). (See Figure 10). Interestingly, low doses were associated with hydroxyproline content and collagen deposition. Although levels showed a significant decrease, the 10 nmol dose resulted in the best survival rate.

[0259] Example 8 Embodiments of the compounds of this disclosure other than Compound 1A and Compound 1B function similarly in application. To support this, other representative embodiments of the compounds described herein were investigated in in vitro studies. Beta.

[0260] Figures 13A-13D show IL-4 at 20 ng / mL, IL-13 at 20 ng / mL, and 5n Measured from human THP-1 cells induced by M2 macrophages with g / mL IL-6. The graph shows data at various marker levels. Next, cells are shown with TLRs having formula IV. 7 Reprogrammed for 48 hours with different nM concentrations of agonist (e.g., compound 2A), q Samples were collected for gene analysis using PCR. mRNA levels of the markers below: CCL18 mRNA level (Figure 12A), IL-1β mRNA levels (Figure 13B), and TNFα levels (Figure 13C), and Figure 13D are shown below. The results of protein analysis after collecting the cell supernatant are shown. Secreted CCL18 protein It was detected by ELISA.

[0261] Figures 13A to 13D show the agonist compounds of the present disclosure having formula IV (e.g., compound 2 A) is the ability to reprogram M2-like macrophages into M1-like macrophages. I evaluated it.

[0262] Mainly, human monocytic (THP-1) cells were expressed in an M2-like manner using the methods and materials described above. The cells were induced into a specific type. Specifically, THP-1 cells were introduced at a density of 60,000 cells / well. The cells were seeded in a well plate. The cells were incubated with 200 nM PMA for 48 hours. Then, by incubating in fresh RPMI medium for 24 hours, the nonpolar compounds are produced. They were differentiated into macrophages. The obtained macrophages were treated with 20 ng / ml of IL-4. IL-13 at 20 ng / ml and IL-6 at 5 ng / ml were incubated for 48 hours. By performing this process, the culture polarized into an M2-like phenotype. It was maintained at 37°C in an incubator.

[0263] Compound 2A reprograms pro-fibrotic macrophages to a lower fibrillation phenotype. To evaluate whether it is possible, we used IL-4, IL-6 plus IL-13. Induced THP-1 cells were incubated with compound 2A at different concentrations, and several lines were observed. mRNA levels of vascularization-promoting markers, namely CCL18, IL-1β, and TNFα. This was investigated using qPCR and ELISA.

[0264] As shown in Figures 13A and 13B, compound 2A (free agent) was incubated for 48 hours. The treatment induces a decrease in the expression of CCL18 and IL-1β, and TLR7 agonists... These THP-1 cells, polarized to promote fibrosis, lead to a lower fibrous phenotype. This suggests that it can actually promote the process. (Note: Figure 13B shows that it has an inhibitory response at low concentrations.) The curve shows a bell shape indicating compound 2A, which exhibits an irritant response at high concentrations, and this is due to a specific drug. This is a common response curve.) Furthermore, the expression of TNFα (anti-fibrotic phenotype marker) was examined. Upon investigation, an increase in its expression was observed (Figure 13C), indicating that THP-1 has fibrosis-promoting properties and is anti- We confirmed that the characteristics shifted towards fibrosis.

[0265] In addition to non-conjugate TLR7 agonism, the conjugate compound of this disclosure also exhibits the same properties. Human THP-1 cells were evaluated as follows: Human THP-1 cells were evaluated as M2-like phenotype according to the method described herein. It is induced into macrophages that have (for example, 20 ng / mL IL-4, 20 ng / m Using IL-13 at 5 ng / mL and IL-6 at 5 ng / mL, then various aspects of the present disclosure described below. The compounds were reprogrammed for 2 hours at different nM concentrations, i.e., formula I and / or Non-conjugate (free drug) TLR7 agonist compounds containing II (data is for compound Folic acid conjugate TLR7 agonists having formula XV (collectively represented as 3A) Compounds (having a releaseable linker) (e.g., compound 3B), folic acid having formula XVII Conjugate TLR7 agonist compounds (with non-release linkers) (e.g., compound Substance 3C), and a folic acid conjugate TLR7 agonist compound having formula XVI (release) (Having a non-linking agent) (e.g., compound 3D). Then, the cells are genetically analyzed by qPCR. Samples were taken for sub-analysis and analyzed for CCL18 (Figure 14A), CD206 (Figure 14B), and IL The relative expression of -1β (Figure 14C) was analyzed.

[0266] Various fibrosis-promoting (M2 phenotype) markers include CCL18, IL-1β, and CD The expression of 206 markers was quantified. As shown in Figures 14A-14C, these markers promote fibrosis. The expression of each marker is as follows: Reduced after administration, compound 3D and compound 3C (both containing non-release linkers) (ru) was the most effective compared to other compounds.

[0267] Figure 15 shows the results after treatment with compound 3A, compound 3B, compound 3C, or compound 3D. Figures 14A-14C show the secreted CCL18 protein levels in each group of THP-1 cells. This shows compound 3A and folic acid-targeted TLR7 compounds (e.g., compound 3B, compound 3 Compounds C and 3D) doubling the secretion of CCL18 in the low concentration range (0.1-10 nM). To regulate.

[0268] Furthermore, the cell supernatant was collected and the secreted CCL18 protein was detected by ELISA. Figure 15 shows compound 3A (free agent) and folic acid targeting compound (compound 3B, compound Substance 3C and compound 3D all exhibit CCL18 in the low concentration range (0.1-10 nM). We confirmed that the secretion was downregulated, and furthermore, compound 1A and compound 1B Similar to the examples described in relation to this, these compounds are M2-like fibers via a similar mechanism. Reprogramming pro-fibrotic macrophages to M1-like anti-fibrotic macrophages in a similar manner. I support the ability to do so.

[0269] Example 9 Repeating the above study (Figures 3A-3F) revealed the same qualitative changes in compound 1B. Only the magnitude of the impact was slightly reduced. Non-targeted TLR7 agonists immediately affected cultured cells. In contrast to entry, its folate-targeted counterpart is folate receptor binding and receptor-mediated This reduced efficacy is due to the fact that it is designed to enter cells only after endocytosis. This was expected. Low molecular weight water-soluble drugs such as compound 1A and compound 1B are expected to be used in many situations. In addition, because it is excreted from the body within 2 hours of injection, it is more physiologically effective for in vivo drug exposure. A suitable in vitro model limits the incubation of cells with drugs to only two hours. Then, the effectiveness of the drug will be examined after a further 46-hour incubation period in the absence of the drug. As shown in Figures 4A-4E, THP-1 cells were incubated with a TLR7 agonist for 2 hours. After that, when the drug-containing medium is replaced with a drug-free medium, compound 1B becomes compound 1A It has been observed to have superior efficacy compared to, in particular, folate targeting in the case of TNFα induction. Conjugation was dramatically improved. This is because folate-targeted TLR7 agonists are acting on folate receptors. Compound 1A was captured by somatic-positive cells, whereas compound 1A was not retained by the same cells. It appears to be folded.

[0270] These data show that compound 1B reprograms pro-fibrotic macrophages in vivo. Folic acid-conjugated drugs should be more effective in molting. (For example, compound 1B) is concentrated in FRβ-expressing macrophages and is dominant throughout the body. Acid receptor-negative cells (for example, compound 1B is impermeable to folate receptor-negative cells) (It is designed to prevent) entry, and therefore does not cause more systemic toxicity. This supports the idea that it has further advantages.

[0271] Furthermore, the above mRNA analysis was found to be produced by IL-4, IL-6, plus IL-13. To accurately reflect the levels of fibrosis-promoting cytokines, stimulate THP-1 cells. Therefore, the concentrations of CCL18 and IL-1β polypeptide in the THP-1 supernatant are controlled by ELI Quantitative analysis was performed by SA assay. As shown in Figures 6A and 6B, compound 1A and chemical When both compounds 1B were incubated with the agonist for 48 hours, CCL1 It induced a decrease in 8 and IL-1β, but the compound was only activated when drug exposure was limited to 2 hours. Item 1B was again found to be superior (see Figures 6C and 6D).

[0272] Example 10 Figure 16 shows at least one embodiment of the compound of this disclosure in a BM mouse model. This describes the vivo research methodology, and the compounds are those having formula XVII (for example, compound 3 C) is the case. Figures 17A and 17B are the LC-MS spectra of compound 3C, and The high purity of the jugate was supported, and no free agents were detected.

[0273] Figures 18A-18F show the results from the mice used in the in vivo study methodology in Figure 16. Existence curve (Figure 18A), weight change (Figures 18B and 18D), cell concentration by BALF ( Figure 17C), living mice (Figure 18E) and all mice (e.g., living mice and Hydroxyproline in mice that died before day 21 (including both mice) (Figure 18F) Contains concentration (μg HP / leaf). 10 compounds having formula XVII (e.g., compound 3C). The nmol concentration dose increased the survival rate of the target mice, and simultaneously increased HP and BALF cells. The number of [unclear] was reduced. Furthermore, the 3 nmol concentration dose showed a measurable benefit in the target mice. It wasn't there.

[0274] Example 11 M2-induced human monocyte-derived macrophages were treated with 100 nM of compound 1A or compound 1B. , administer continuously for 48 hours, or initially in the presence of FA-glucosamine (a competitor) Either treat for 2 hours in the absence of the drug, followed by 46 hours in the absence of the drug (2 + 46 hours). One of the following was performed: As shown in Figure 19, the fibrosis-promoting marker, Arg1 (Figure 19A) ), mRNA levels of CD206 (Figure 19B) and CD163 (Figure 19C), and Secreted pro-fibrotic CCL18 (Figure 19D) and anti-fibrotic cytokine, CXCL1 Protein levels of 0 (Figure 19E) and IL-6 (Figure 19F) (n=3, technical replication) Next, it was determined that the changes in both sets of cytokines were due to excessive FA-glucosamine (2 Inhibited by blockade of unoccupied folate receptors (with competition) for +46 hours. This data shows that FA-glucosamine has excessive downregulation of biomarkers. The fact that it was blocked by a competitor supports the idea that compound 1B binds to the folate receptor.

[0275] Example 12 Healthy mice were given 10 nmol of compound 1A (round) or compound 1B (square) at their tails. The drug was administered by pulse injection, and peripheral blood was collected at the designated time after the injection. (Figure 20A~C) Plasma IL -6 (Figure 20A), IFNα (Figure 20B), and TNFα (Figure 20C) measurements (n=3) (Figures 20D-F). IL-6 (Figure 20D), IFNα (Figure 20E), and TNFα ( The effect of drug concentration on plasma levels in Figure 20F) is shown at 1.5 hours and 1 hour after treatment, respectively. or determined in 1 hour (n=2) (Figure 20G). Compound 1A was found in healthy mice. It stimulates systemic cytokine release, but compound 1B does not. Furthermore, compound 1B These data stimulate the release of inflammatory cytokines at a dose lower than half that of compound 1A. TLR7 agonists target lung macrophages using folate receptor-targeting ligands. If targeted, in order to reprogram fibrotic lung macrophages into a fibrotic-promoting state This suggests that it can be safely implemented.

[0276] Example 13 Sections of healthy and fibrous lung tissue, the same as those shown in Figure 6, are shown with DAPI (nucleus, blue). Anti-F4 / 80 (macrophage, red), and anti-CD206 (M2 macrophage mer). Dye with (green) and use a Leica Versa 8-hole slide as described in the method. Images were acquired using an IdScanner (n=2). Scale bar, 100 μm. Differences between treatment groups. This indicates that compound 1B elicits a potent pro-fibrotic response in vivo.

[0277] Various embodiments of compounds, compositions, and methods are described in considerable detail herein. However, the embodiments described herein are provided merely as non-limiting examples. Many variations and modifications of the embodiments will be apparent to those skilled in the art in light of this disclosure. Therefore, various changes and modifications may be made without departing from the scope of this disclosure. Those skilled in the art will understand that equivalents can be substituted for those elements. In fact, this disclosure is not intended to be exhaustive or excessively restrictive. The scope is defined by the attached claims and their equivalents.

[0278] Furthermore, many of the examples provided herein use mouse models, but mouse The gene expression patterns in Dell show a very strong correlation with those in the human state. This indicates that many pathways are generally controlled by multiple conditions in humans and mice. This will be understood by those skilled in the art. Therefore, gene development in mouse models Current patterns and disease progression are characteristic of human pathogenesis, particularly in relation to inflammatory diseases and cancer. This method strictly reproduces the results, and therefore the examples described herein are based on human data and specific diseases. It supports the correlation between state and application.

[0279] Therefore, this description and the attached claims do not contain anything that would be obvious to a person skilled in the art based on this disclosure. It is intended that all corrections and changes are included.

Claims

1. A compound represented by the following formula, Q-L-T During the ceremony, Q is a radical of the folate receptor binding ligand, L is a linker, T is a Toll-like receptor (TLR) agonist or a pharmaceutically acceptable salt thereof. A compound that is CAL.

2. The compound according to claim 1, wherein the linker is a non-release linker.

3. The aforementioned non-dischargeable linker is as follows: 【Chemistry 1】 Represented by, During the ceremony, n is between 1 and 30. The compound according to claim 2, wherein w is 0 to 5.

4. The aforementioned TLR agonist is a Toll-like receptor 7 (TLR7) agonist. A compound listed in any one of the requested items.

5. The radical of the TLR agonist has a structure represented by formula X; 【Chemistry 2】 During the ceremony, R １ Ha-NH ２ or -NH-R １Ｘ And, R ２ H, alkyl, alkenyl, alkynyl, alicyclic, aryl, biaryl, he Terroraryl, -NH-R ２Ｘ , -O-R ２Ｘ , -S-R ２Ｘ , 【Transformation 3】 And, R １Ｘ 、R ２Ｘ 、and R ２Ｙ each is independently H, alkyl, alkenyl, alk Selected from the group consisting of nyl, alicyclic, aryl, biaryl, and heteroaryl, 【Chemistry 4】 It is a 3-10 member nitrogen-containing non-aromatic monocyclic or bicyclic heterocycle, R ３ -OH, -SH, -NH ２ , or -NH-R １Ｘ The prior claim is A compound listed in any one of the items.

6. The radical of the TLR agonist has a structure represented by formula XX, 【Transformation 5】 During the ceremony, R １ is, -NH ２ or -NH-R １Ｘ And, R ２ H, alkyl, alkenyl, alkynyl, alicyclic, aryl, biaryl, he Terroraryl, -NH-R ２Ｘ , -O-R ２Ｘ , -S-R ２Ｘ , 【Transformation 6】 And, R １Ｘ , R ２Ｘ , and R ２Ｙ Each of these can independently be H, alkyl, alkenyl, or alkyl Selected from the group consisting of nyl, alicyclic, aryl, biaryl, and heteroaryl, 【Transformation 7】 It is a 3-10 member nitrogen-containing non-aromatic monocyclic or bicyclic heterocycle, X is CH, CR ２ The compound according to any one of claims 1 to 4, wherein the compound is N.

7. The radical of the TLR has a structure represented by formula XXX. The compound according to claim 6, which is a strike. 【Transformation 8】

8. Linker L between the targeting moiety and the immunomodulator or its pharmaceutically acceptable salt. ｎ of Furthermore, the linker L ｎ However, to avoid the release of the free form of the TLR7 agonist A compound according to any one of the prior claims, wherein n is an integer less than or equal to 50.

9. Linker L ｎ The linker L ｎ However, polyethylene glycol (PEG) or The PEG derivative comprises an integer n selected from the range of 1 to 32, and the folate receptor Any one of the prior claims, wherein the radical of the binding ligand is a folate receptor β-binding ligand. The compounds listed in the section.

10. The compound according to claim 1, wherein the compound has the structure represented below. 【Chemistry 9】

11. The compound according to claim 1, wherein the compound has the structure represented below. 【Chemistry 10】

12. The compound according to claim 1, wherein the compound has the structure represented below. 【Chemistry 11】

13. The compound according to claim 1, wherein the compound has the structure represented below. 【Chemistry 12】

14. The TLR7 agonist has a structure represented by formula XX, 【Chemistry 13】 During the ceremony, R １ Ha-NH ２ or -NH-R １Ｘ And, R ２ H, alkyl, alkenyl, alkynyl, alicyclic, aryl, biaryl, he Terroraryl, -NH-R ２Ｘ , -O-R ２Ｘ , -S-R ２Ｘ , 【Chemistry 14】 And, R １Ｘ , R ２Ｘ , and R ２Ｙ Each of these can independently be H, alkyl, alkenyl, or alkyl Selected from the group consisting of nyl, alicyclic, aryl, biaryl, and heteroaryl, 【Chemistry 15】 It is a 3-10 member nitrogen-containing non-aromatic monocyclic or bicyclic heterocycle, X is CH, CR ２ A pharmaceutical composition comprising the compound according to claim 4, which is either N or N.

15. A method for treating a subject suffering from a fibrotic disease or cancer, wherein the method is The target cells are subjected to the compound described in any one of claims 1 to 13, or the compound described in claim 14. The process involves contacting at least one compound containing the pharmaceutical composition described above with the TLR The radical of the gonist or a pharmaceutically acceptable salt thereof is the jaw of TLR7, 8, or 9. Methods, including stylists.

16. Folate ligands attached to Toll-like receptor (TLR) agonists via linkers or a compound containing a functional fragment or analog thereof, wherein the TLR agonist is as follows Having the formula or a pharmaceutically acceptable salt thereof, 【Chemistry 16】 During the ceremony, R １ It is an amine group, R ２ It is a single bond -NH-, R ３ is H, alkyl, hydroxyl group, or any other substituent thereof. X is CH ２ , NH, O, or S, The linker is R １ 、 R ２ , or R ３ Compounds that adhere to the surface.

17. The linker is a polyethylene glycol (PEG) linker or a PEG derivative phosphorus Includes car, R ３ Is it a non-release linker that has adhered to it, or R １ , R ２ or R ３ Adhering The present invention is either a releaseable linker or any one of claims 1 to 13. A pharmaceutical composition comprising the compound described or the composition described in claim 16.

18. The aforementioned pharmaceutically acceptable salts are hydrobromide, citrate, trifluoroacetate, and as Corbic acid, hydrochloride, tartrate, triflate, maleate, mesylate, formate, vinegar Selected from salts or fumarates, any one of claims 1 to 13 or claim 16. The compound described in item 1, or the pharmaceutical composition described in claim 14.

19. A method for preventing or treating a fibrous disease state, Cells are attached to folate ligands or functional fragments or analogs via linkers. A compound comprising an immunomodulatory substance or a pharmaceutically acceptable salt thereof, and By bringing the immunomodulator or a pharmaceutically acceptable salt thereof into contact with the pattern A method comprising targeting a receptor for recognition of , and bringing it into contact with, and

20. Does the immunomodulator contain a Toll-like receptor (TLR) agonist of formula X or XX? , or a pharmaceutically acceptable salt of formula X or XX, 【Chemistry 17】 In equations X and XX, R １ is, -NH ２ or -NH-R １Ｘ And, R ２ H, alkyl, alkenyl, alkynyl, alicyclic, aryl, biaryl, he Terroraryl, -NH-R ２Ｘ , -O-R ２Ｘ , -S-R ２Ｘ , [Chemistry 18] And, 【Chemistry 19】 It is a 3-10 member nitrogen-containing non-aromatic monocyclic or bicyclic heterocycle, In equation X, R ３ -OH, -SH, -NH ２ or -NH-R １Ｘ And, In equation XX, X is either CH or N. R １Ｘ , R ２Ｘ , and R ２Ｙ Each of these can independently be H, alkyl, alkenyl, or alkyl Selected from the group consisting of nyl, alicyclic, aryl, biaryl, and heteroaryl. The method according to claim 19.

21. The aforementioned cells are target cells that are experiencing or at risk of experiencing a fibrous disease state. Including the above, contacting the cells with at least one compound is considered to be a therapeutically effective amount of the above less Claim 19 further comprises administering or applying at least one compound to the subject. Method of loading.

22. The subject is a patient who has experienced idiopathic pulmonary fibrosis, and the subject is at least one compound However, the subject is administered intravenously, intramuscularly, intraperitoneally, locally, or by inhalation. The method according to claim 21.

23. The aforementioned fibrotic disease state may be idiopathic pulmonary fibrosis, or affect the liver, skin, bladder, heart, pancreas, prostate, etc. The method according to claim 21, comprising glandular or renal fibrotic disease.

24. Obtaining a sample from the aforementioned subject, or having obtained a sample, The method of quantifying the expression level of one or more biomarkers in the sample, Each of one or more biomarkers is a chemokine (C-C motif) ligand 18 (CC L18), Arginase 1 (Arg1), Matrix Metalloproteinase 9 (MMP 9) Metalloproteinase 3 (TIMP3), Interleukin 1 beta (IL-1β) ), hydroxyproline, collagen, platelet-derived growth factor (PDGF), transformation growth Factor-beta (TGFβ), folate receptor beta (FRβ), tumor necrosis factor alpha (TN) Fα), interferon-gamma (IFN-γ), mannose receptor (CD206), Antigen group 163 (CD163), differentiated antigen group 86 (CD86), interleukin 6 (I L-6), chemokine 10 (CXCL10), and immune interferon (IFNα) Selected from the group consisting of, The expression level of each of the one or more biomarkers in the sample is compared with the control. This involves comparing the expression levels of such biomarkers, CCL18, Arg1, MMP9, TIMP3, IL-1β, PDGF, TGFβ, C D206, CD163, FRβ, hydroxyproline, or collagen are the control Compared to the aforementioned expression levels, it is upregulated, or TNFα, IF One or more of N-γ, IL-6, CXCL10, IFNα, and CD86 are the same as the above. Compared to the aforementioned expression levels of irradiate, it is downregulated or not expressed at all. In such cases, a therapeutically effective amount of an unconjugated agonist or inhibitor is administered to the subject. The method according to claim 21, further comprising administering to or having administered.

25. The folate ligand or its functional fragment or analog is specific to the folate receptor β. The method according to claim 21, wherein the cell binds to the folate receptor β.

26. Immunomodulators that target cellular pattern recognition receptors or their pharmaceutically acceptable salts A compound comprising a targeted portion attached to a folic acid ligand or the A compound containing a functional fragment or analogue of [the substance].

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