Application of small molecule compound in preparation of CXCR7 agonist

Abstract

The invention relates to the field of compounds, in particular to application of a small molecule compound in preparation of a CXCR7 agonist. The invention provides an application of a small molecule compound or a pharmaceutically acceptable salt thereof in preparation of a CXCR7 agonist, and is characterized in that the small molecule compound comprises one or more of JMS-17-2, rupatadine, ibutamolam mesylate, asenapine, alverine and vilanterol. The small molecule compound can specifically activate an intracellular beta-arrest signal channel instead of being coupled with a G protein signal channel, so that the small molecule compound can be used as a novel CXCR7 agonist.

Description

[0001] This application claims priority to Chinese Invention Patent Application 2024112522951, “A CXCR7 Agonist”, filed on September 6, 2024, which is incorporated by reference in its entirety. TECHNICAL FIELD

[0002] The present application relates to the field of compounds, in particular to the use of a small molecule compound in the preparation of a CXCR7 agonist. BACKGROUND

[0003] Organ fibrosis is a major pathological feature of many diseases, and about one-third of global disease causes are attributed to fibrosis. Pulmonary fibrosis leads to respiratory failure and death in patients, with an average survival period lower than lung cancer. Existing clinical drugs, such as nintedanib and pirfenidone, cannot effectively reverse the progression of pulmonary fibrosis. Promoting the regeneration of fibrotic lung tissue is expected to restore the structure of damaged alveoli and blood vessels, providing a strategy for the treatment of various diseases, including pulmonary fibrosis.

[0004] The lung is a highly vascularized organ, with capillary endothelial cells located between the blood circulation and epithelial and interstitial cells. In addition to performing gas exchange functions, vascular endothelial cells interact with adjacent epithelial, interstitial, and blood cells through paracrine “Angiocrine” factors to regulate lung regeneration and fibrosis. CXCR7, which is specifically highly expressed in lung capillaries, can serve as a key vascular microenvironment target to coordinate lung regeneration and fibrosis processes. In the vascular microenvironment of fibrotic lung tissue, CXCR7 signaling is inhibited, leading to pro-fibrotic Angiocrine signals that stimulate perivascular fibroblasts, thereby activating the fibrotic response and inhibiting lung regeneration. By targeting CXCR7 in the lung vascular microenvironment, the receptor can be activated, reprogramming the Angiocrine signals in the vascular microenvironment, inhibiting fibroblast activation, and stimulating alveolar stem cell viability, thereby promoting the repair and regeneration of fibrotic lung tissue.

[0005] G protein-coupled receptors (GPCRs) are a superfamily of membrane proteins with seven transmembrane helices (7TMs), which is the largest receptor superfamily discovered so far, and widely involved in various important life activities such as reproduction, growth, development and perception, thus attracting much attention in drug development. As of 2017, about 34% of all drugs approved by the US FDA target GPCRs. The traditional receptor-ligand interaction model suggests that ligands can mediate downstream intracellular signaling pathways after binding to receptors. However, in the experimental study of muscarinic cholinergic receptor agonists, it was found that some ligands can induce receptors to selectively bind to specific G protein subunits, so that the intracellular signal is biased to a certain pathway among numerous downstream pathways, which is known as "signal bias". Subsequent studies have shown that this bias not only reflects the selective binding of different G protein subunits and the exertion of completely different physiological functions, but also reflects the selective bias of G protein pathways or G protein-independent β - one of the arrestin (β-arrestin) pathways, ultimately leading to completely different biological effects.

[0006] That is, GPCRs have the property of "signal bias", i.e., bias towards G protein and β-arrestin activation. CXCR7 can specifically activate the intracellular β-arrestin signaling pathway without coupling to the G protein signaling pathway. This biased β-arrestin signal transduction property helps to design CXCR7 agonists, so that they can avoid activating the G protein signaling pathway when treating organ fibrosis (e.g., pulmonary fibrosis), thereby reducing side effects. However, there is no small molecule drug targeting CXCR7 that has been clinically approved, and there is a lack of efficient CXCR7 agonists. SUMMARY

[0007] In a first aspect, the present application provides the use of a small molecule compound or a pharmaceutically acceptable salt thereof in the preparation of a CXCR7 agonist, characterized in that the small molecule compound comprises one or more of JMS-17-2, rupatadine, ibuprofen mesylate, asenapine, alverine and vilanterol.

[0008] As used herein, "agonist" refers to a substance that promotes (e.g., induces, causes, enhances, or increases) the biological activity or effect of another molecule.

[0009] As used herein, "CXCR7 agonist" refers to a substance that fully or partially activates the CXCR7 receptor. In some embodiments, the CXCR7 agonist is a compound. The term "CXCR7 agonist" and specific CXCR7 agonists described herein are also intended to encompass their salt forms.

[0010] As used herein, the term "pharmaceutically acceptable salt" refers to a derivative of a compound in which the parent compound is modified by converting an existing acid or base moiety into its salt form. Pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues such as amines; alkali or organic salts of acidic residues such as carboxylic acids. The pharmaceutically acceptable salts include the conventional nontoxic salts of the parent compound, for example, from non-toxic inorganic or organic acids. The pharmaceutically acceptable salts can be synthesized from the parent compound which contains a basic or acidic moiety by conventional chemical methods. Generally, such salts can be prepared by reacting the free acid or base forms of these compounds with a stoichiometric amount of the appropriate base or acid in water or in an organic solvent or in a mixture of the two. Examples of suitable inorganic acids include hydrochloric, sulphuric, phosphoric or hydrobromic acid, while examples of suitable organic acids include carboxylic or sulphonic acids, such as formic, acetic, tartaric, lactic, propionic, glycolic, malonic, maleic, fumaric, tannic, succinic, alginic, benzoic, 2-phenoxybenzoic, 2-acetoxybenzoic, cinnamic, mandelic, citric, maleic, salicylic, trifluoroacetic, 3-aminosalicylic, ascorbic, aconitic, nicotinic, isonicotinic, oxalic, gluconic, amino acids, methanesulphonic, ethanesulphonic, 2-hydroxyethanesulphonic, ethane-1, 2-disulphonic, benzenesulphonic, 4-methylbenzenesulphonic or naphthalene-2-sulphonic acid. Examples of suitable inorganic bases include sodium hydroxide, potassium hydroxide and ammonia, while examples of suitable organic bases include amines, such as tertiary amines, for example, trimethylamine, triethylamine, pyridine, N,N-dimethylaniline, quinoline, isoquinoline, a-methylpyridine, β-methylpyridine, γ-methylpyridine, quinaldine or pyrimidine.

[0011] In some embodiments, the small molecule compound is JMS-17-2, ibuprofen mesylate or rupatadine.

[0012] In some embodiments, the small molecule compound is JMS-17-2, ibuprofen mesylate or rupatadine.

[0013] In some embodiments, the small molecule compound is JMS-17-2, ibuprofen mesylate or rupatadine.

[0014] As used herein, "fibrosis" refers to the excessive or unregulated deposition of extracellular matrix components and abnormal repair processes in tissues or organs following injury, such as skeletal muscle, cardiac muscle, liver, lung, kidney and pancreas.

[0015] In some embodiments, the organ fibrosis comprises one or more of pulmonary fibrosis, liver fibrosis and kidney fibrosis.

[0016] In a third aspect, the present application also provides a use of a small molecule compound or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating and / or preventing a pulmonary disease, wherein the small molecule compound comprises one or more of JMS-17-2, rupatadine, ibuprofen mesylate, asenapine, alverine and vilanterol.

[0017] In some embodiments, the small molecule compound is JMS-17-2, ibuprofen mesylate or rupatadine.

[0018] As used herein, "pulmonary disease" refers to a disease occurring in the lung, including a disease of the lung itself or a pulmonary manifestation of a systemic disease. In some embodiments, the pulmonary disease comprises one or more of pulmonary fibrosis, interstitial lung disease, chronic obstructive pulmonary disease and lung injury.

[0019] In some embodiments, the lung injury comprises a post-infection or post-surgery lung injury.

[0020] In a fourth aspect, the present application also provides a use of a small molecule compound or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating and / or preventing an organ injury, wherein the small molecule compound comprises one or more of JMS-17-2, rupatadine, ibuprofen mesylate, asenapine, alverine and vilanterol.

[0021] In some embodiments, the organ comprises one or more of lung, liver and kidney.

[0022] In some embodiments, the organ injury comprises one or more of lung injury, liver injury and kidney injury.

[0023] In some embodiments, the use comprises providing a protective effect to the organ after the organ injury.

[0024] In a fifth aspect, the present application also provides a use of motupirinQ in the manufacture of a CXCR7 agonist.

[0025] In a sixth aspect, the present application also provides a compound, wherein the compound is a compound represented by Formula I as shown below, or a stereoisomer, a geometric isomer, a tautomer, a hydrate, a solvate, and a pharmaceutically acceptable salt or a prodrug thereof: (I), wherein L is (C1-C6)alkyl; A is a saturated 3-6 membered aliphatic ring, heterocyclic ring, bridged ring or spiro ring; R is -C(=O)NHR x or -C(=O)ORx or substituted aryl or arylhetero group; R x is a hydrogen atom, a methyl group, an ethyl group, a propyl group, an isopropyl group, a butyl group, or a tert-butyl group.

[0026] In some embodiments, the compound is a compound of Formula IV (IV) wherein Rz is a substituent of at least one of ortho-substitution, meta-substitution, para-substitution, the substituent being selected from the group consisting of halogen, methoxy, methyl, ethyl, propyl, isopropyl, butyl, tert-butyl, trifluoromethyl, and trifluoromethoxy.

[0027] In some embodiments, the halogen is selected from the group consisting of fluorine, bromine, and chlorine.

[0028] In some embodiments, Rz is halogen or methoxy of ortho-substitution, meta-substitution, and para-substitution.

[0029] In some embodiments, the halogen is selected from the group consisting of fluorine, bromine, and chlorine. In some embodiments, the compound is a compound of Formula II (II).

[0030] In a seventh aspect, the present application also provides a compound, characterized in that the compound is a compound represented by Formula III as shown below, or a stereoisomer, a geometric isomer, a tautomer, a hydrate, a solvate, and a pharmaceutically acceptable salt or a prodrug thereof: (III).

[0031] In an eighth aspect, the present application also provides use of the above-mentioned compound in preparation of a CXCR7 agonist.

[0032] In a ninth aspect, the present application also provides use of the above-mentioned compound in preparation of a medicament for treating and / or preventing organ fibrosis.

[0033] In some embodiments, the organ fibrosis comprises one or more of pulmonary fibrosis, liver fibrosis, and kidney fibrosis.

[0034] In some embodiments, the symptoms of the organ fibrosis comprise an increase in the level of one or more of the following molecules: α-smooth muscle actin (α-SMA), collagen I, hydroxyproline.

[0035] In some embodiments, the medicament is used to reduce one or more of the following symptoms: degree of organ fibrosis, degree of organ damage, degree of collagen deposition, degree of fibroblast activation.

[0036] As used herein, “alkyl” includes branched and straight-chain saturated aliphatic hydrocarbon groups having a specified number of carbon atoms. For example, (C1-C6)alkyl is intended to include C1, C2, C3, C4, C5, and C6 alkyl groups. Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl-sec-butyl, tert-butyl, n-pentyl, sec-pentyl, neopentyl, and n-hexyl. In some cases, straight-chain or branched alkyl groups have six or fewer carbon atoms in their main chain (e.g., straight-chain C1-C6, branched C3-C6), and in others, straight-chain or branched alkyl groups have four or fewer carbon atoms. Similarly, cycloalkyl groups have three to eight carbon atoms in their ring structure, and in others, cycloalkyl groups have five or six carbon atoms in their ring structure. Preferred are (C1-C6)alkyl groups, particularly ethyl, methyl, isopropyl, isobutyl, n-pentyl, n-hexyl, and cyclopropylmethyl.

[0037] As used herein, “substitution” means the selection of any one or more hydrogen atoms on a specified atom to be replaced by an indicated group, provided that the normal valence of the specified atom does not exceed a certain value and the substitution results in a stable compound.

[0038] As used herein, “aryl” refers to a monocyclic or polycyclic aromatic hydrocarbon (e.g., having 2, 3, or 4 fused rings), such as phenyl, naphthyl, anthracene, or phenanthrene. In some cases, aryl groups have 6–20 carbon atoms. As used herein, the term “aromatic heterocycle” refers to an aromatic heterocycle having at least one heteroatom ring member such as O, S, or N atom. Aromatic heterocycle groups include monocyclic and polycyclic systems (e.g., having 2, 3, or 4 fused rings). The cyclic N atom in an aromatic heterocycle group can also be oxidized to form an N-oxygen moiety. Examples of aromatic heteroyl groups include pyridinyl, N-oxopyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, triazinyl, furanyl, quinolinyl, isoquinolinyl, thiopheneyl, imidazolinyl, thiazolyl, indoleyl, pyrroleyl, oxazolyl, benzofuranyl, benzothiopheneyl, benzothiazolyl, isoxazolyl, pyrazolyl, triazolyl, tetrazolyl, indoleyl, 1,2,4-thiophenediazolyl, isothiazolyl, benzothiopheneyl, purine, carbazoleyl, benzimidazolinyl, and dihydroindoleyl. In some cases, aromatic heteroyl groups have 1 to about 20 carbon atoms, and in others about 3 to 20 carbon atoms. In some cases, aromatic heteroyl groups contain 3 to about 14, 3 to about 7, or 5 to 6 cyclic atoms. In some cases, aromatic heteroyl groups have 1 to about 4, 1 to about 3, or 1 to 2 heteroatoms.

[0039] In a tenth aspect, the present invention also provides the use of the above-described compounds in the preparation of medicaments for treating and / or preventing lung diseases.

[0040] In some embodiments, the lung disease includes one or more of pulmonary fibrosis, interstitial lung disease, chronic obstructive pulmonary disease, and lung injury.

[0041] In some embodiments, the lung injury includes lung injury following infection or surgery.

[0042] In an eleventh aspect, the present invention also provides the use of the above-mentioned compounds in the preparation of medicaments for treating and / or preventing organ damage.

[0043] In some embodiments, the organ includes one or more of the lungs, liver, and kidneys.

[0044] In some embodiments, the organ injury includes one or more of lung injury, liver injury, and kidney injury.

[0045] In some embodiments, the use includes providing protection to the organ after injury to the organ.

[0046] In a twelfth aspect, the present invention also provides the use of the compound represented by Formula IV in the preparation of CXCR7 agonists. (IV) Wherein Rz is a substituent of at least one of ortho-substitution, meta-substitution, and para-substitution, wherein the substituent is selected from the group consisting of halogen, methoxy, methyl, ethyl, propyl, isopropyl, butyl, tert-butyl, trifluoromethyl, and trifluoromethoxy.

[0047] In some embodiments, the halogen is selected from the group consisting of fluorine, bromine, and chlorine.

[0048] In some embodiments, Rz is a halogen or methoxy group that is ortho-substituted, meta-substituted, or para-substituted.

[0049] In some embodiments, the halogen is selected from the group consisting of fluorine, bromine, and chlorine.

[0050] In a thirteenth aspect, the present invention also provides the use of the compound represented by Formula IV in the preparation of medicaments for treating and / or preventing organ fibrosis.

[0051] In some embodiments, the organ fibrosis includes one or more of pulmonary fibrosis, liver fibrosis, and kidney fibrosis.

[0052] In a fourteenth aspect, the present invention also provides the use of the compound represented by Formula IV in the preparation of a medicament for treating and / or preventing lung diseases.

[0053] In some embodiments, the lung disease includes one or more of pulmonary fibrosis, interstitial lung disease, chronic obstructive pulmonary disease, and lung injury.

[0054] In some embodiments, the lung injury includes lung injury following infection or surgery.

[0055] In a fifteenth aspect, the present invention also provides the use of the compound represented by Formula IV in the preparation of a medicament for treating and / or preventing organ damage.

[0056] In some embodiments, the organ includes one or more of the lungs, liver, and kidneys.

[0057] In some embodiments, the organ injury includes one or more of lung injury, liver injury, and kidney injury.

[0058] In some embodiments, the use includes providing protection to the organ after injury to the organ.

[0059] In some embodiments, the dosage range of the compound is about 1 mg / kg to about 40 mg / kg, 0.1 mg / kg to about 4 mg / kg, or 0.01 mg / kg to about 0.40 mg / kg. In some embodiments, the dosage range of the compound is about 1, 2, 3, 4, 5, 6, 7, 8, or 9 mg / kg to about 10, 20, 30, or 40 mg / kg; about 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 mg / kg to about 20, 30, or 40 mg / kg; about 20, 21, 22, 23, 24, 25, 26, 27, 28, or 29 mg / kg to about 30 or 40 mg / kg; about 30, 31, 32, 33, 34, 35, 36, or 37 mg / kg. From about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, or 0.9 mg / kg to about 1, 2, 3, or 4 mg / kg; from about 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, or 1.9 mg / kg to about 2, 3, or 4 mg / kg; from about 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, or 2.9 mg / kg to about 3... Or 4 mg / kg; or from about 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8 or 3.9 mg / kg to about 4 mg / kg; from about 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09 mg / kg to about 0.10, 0.20, 0.30 or 0.40 mg / kg; from about 0.10, 0.11, 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0. 18 or 0.19 mg / kg to about 0.20, 0.30 or 0.40 mg / kg; about 0.20, 0.21, 0.22, 0.23, 0.24, 0.25, 0.26, 0.27, 0.28 or 0.29 mg / kg to about 0.30 or 0.40 mg / kg; or about 0.30 mg / kg to about 0.30, 0.31, 0.32, 0.33, 0.34, 0.35, 0.36, 0.37, 0.38 or 0.39 mg / kg to about 0.40 mg / kg.

[0060] Compared with the prior art, the beneficial effects of the present invention include at least the following aspects: Previous studies have found that CXCR7 signaling is suppressed in the vascular microenvironment of fibrotic lung tissue, leading to pro-fibrotic angiocrine signaling, which stimulates perivascular fibroblasts, thereby activating the fibrotic response and inhibiting lung regeneration. Targeting CXCR7 in the lung vascular microenvironment to activate this receptor could reprogram the angiocrine signaling in the vascular microenvironment, inhibiting fibroblast activation and stimulating alveolar stem cell activity, thereby promoting the repair and regeneration of fibrotic lung tissue. Even so, highly effective CXCR7 agonists are currently lacking.

[0061] This invention identified six small molecule compounds (namely JMS-17-2, rupatadine, ibuprofen mesylate, asenapine, arvilin, and vilanterol) that can specifically activate the intracellular β-arrestin signaling pathway without coupling to the G protein signaling pathway, thus serving as novel CXCR7 agonists. Although all six small molecule compounds target GPCRs, their known uses are as follows: JMS-17-2 is reported to be a selective CX3CR1 antagonist that can inhibit the metastasis and colonization of breast cancer cells; rupatadine is a histamine H1 receptor antagonist and platelet-activating factor inhibitor; ibuprofen mesylate is a selective ghrelin receptor agonist and growth hormone secretagogue (GHSR) that mimics the growth hormone (GH) stimulation effect of the endogenous hormone ghrelin; asenapine is an atypical antipsychotic with central nervous system depressant, antihistamine, and anti-serotonin activities; arvirine is a synthetic papaverine derivative and a selective smooth muscle relaxant; and vilanterol is a novel β2-receptor agonist. In other words, the activation of CXCR7 by these six small molecule compounds has not been reported. Therefore, as CXCR7 agonists, these six small molecule compounds could be further used for the treatment and / or prevention of organ fibrosis, lung diseases, and / or organ damage. Attached Figure Description

[0062] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. In all the drawings, similar elements or parts are generally identified by similar reference numerals. The elements or parts in the drawings are not necessarily drawn to scale. Obviously, the drawings described below are some embodiments of the present invention, and those skilled in the art can obtain other drawings based on these drawings without any creative effort.

[0063] Figure 1 The figure shows the results of discovering novel CXCR7 agonists based on the PRESTO-Tango high-throughput drug screening platform; Figure 2A technology roadmap for designing novel small molecule agonists of CXCR7; Figure 3 The results of testing the activation potential of CXCR7 by cpd-2-3, cpd-1-3, cpd-1-5 and cpd-2-2 using the PRESTO-Tango system are shown in the figure (results are expressed as mean ± SEM (n=3)). Figure 4 The results of in vivo verification of the therapeutic effects of four new compounds on pulmonary fibrosis are shown in the figure. Figure 5 The results of different doses of cpd-2-3 on the therapeutic effects of pulmonary fibrosis are shown in the figure. Figure 6 The image shows the results of CPD-2-3 inhibiting pulmonary fibrosis. Figure 7 The synthesis route for cpd-2-3; Figure 8 This is the synthesis route for cpd-1-3. Detailed Implementation

[0064] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some, not all, of the embodiments of the present invention. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are within the scope of protection of the present invention.

[0065] In this document, "and / or" includes any and all combinations of one or more of the listed related items.

[0066] In this article, "multiple" means two or more, that is, it includes two, three, four, five, etc.

[0067] It should be noted that, in this document, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such a process, method, article, or apparatus. Unless otherwise specified, an element defined by the phrase "comprising one..." does not exclude the presence of other identical elements in the process, method, article, or apparatus that includes that element.

[0068] As used in this specification, the term "about" typically means + / -5% of the value, more typically + / -4% of the value, more typically + / -3% of the value, more typically + / -2% of the value, even more typically + / -1% of the value, and even more typically + / -0.5% of the value.

[0069] In this specification, certain embodiments may be disclosed in a range-bound format. It should be understood that this "range-bound" description is merely for convenience and brevity and should not be construed as a rigid limitation on the disclosed range. Therefore, the description of a range should be considered as having specifically disclosed all possible subranges and the individual numerical values ​​within those ranges. For example, a description of the range 1-6 should be considered as having specifically disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6, etc., and the individual numbers within those ranges, such as 1, 2, 3, 4, 5, and 6. This rule applies regardless of the breadth of the range.

[0070] Detailed description of the attached diagram: Figure 1 A. Roadmap for identifying CXCR7 agonists using the PRESTO-Tango high-throughput drug screening platform. The GPCR drug library contains a total of 1132 compounds. BC. Eight compounds capable of activating CXCR7 were identified using the PRESTO-Tango system. Results are presented as mean ± SEM. n= 3).

[0071] Figure 4 Results from hydroxyproline content assay (A), Masson staining (B), Sirius red staining (C), H&E staining (D), and Collagen I and α-SMA immunofluorescence staining (EF) showed that cpd-2-3 had the most significant therapeutic effect on pulmonary fibrosis, followed by cpd-1-3. cpd-1-5 and cpd-2-2 had no significant effect on pulmonary fibrosis. Results are expressed as mean ± SEM (n=5).

[0072] Figure 5 The therapeutic effects of 2 mg / kg, 5 mg / kg, and 10 mg / kg cpd-2-3 on pulmonary fibrosis were evaluated by hydroxyproline content assay (A), Masson staining (B), Sirius red staining (C), H&E staining (D), and Collagen I and α-SMA immunofluorescence staining (EF). Results are expressed as mean ± SEM (n=5).

[0073] Figure 6CPD-2-3 targets CXCR7 in endothelial cells, reprogramming the angiocrine signaling pathway in the vascular microenvironment to promote lung regeneration and inhibit fibrosis. Lung vascular endothelial cells were isolated using magnetic bead sorting technology, and the expression changes of angiocrine factors, including Hgf, Aplin, Mmp14, Dll4, Kit, Egfl7, Tgfb1, Pdgfb, Igfbp7, Ctgf, Adamts1, and Tgfb2d, were detected. Results are expressed as mean ± SEM (n=3).

[0074] Example 1 To develop novel and highly effective CXCR7 agonists, this embodiment employed the PRESTO-Tango high-throughput drug screening platform, combined with a GPCR small molecule drug library, to screen for small molecule compounds with the potential to activate CXCR7. PRESTO-Tango utilizes a luciferase reporter system to detect the activation of the CXCR7-coupled β-arrestin signaling pathway by small molecule drugs. This is based on the principle that ligands bind to specific G protein-coupled receptors (GPCRs), thereby activating the downstream β-arrestin signaling pathway. This process is mediated by the recruitment of a protease-tagged repressor protein (β-arrestin) to the activated receptor. The protease-tagged repressor protein cleaves the protease site fused to the C-terminus of the GPCR, releasing a non-natural transcription factor. This transcription factor immediately enters the cell nucleus, bypassing additional signal transduction intermediates. Based on the activation of the reporter gene, the degree of interaction between the target receptor and the protease-tagged repressor protein can be quantitatively determined, agonist-mediated receptor activation can be monitored, and the relative potency of different compounds can be evaluated. When CXCR7 on the cell membrane is activated by small molecule drugs, it can further activate the intracellular β-arrestin signaling pathway and stimulate luciferase expression. Figure 1 A).

[0075] Using this system, this embodiment tested 1132 compounds in the GPCR drug library one by one and identified 8 compounds that could activate CXCR7. Figure 1 B. Figure 1 C), namely VUF11207, AMD3100, JMS-17-2, Rupatadine, MK-0667, Asenapine, Alverine, and Vilanterol. Despite the EC of S9665 (Motixafortide). 50 The value was low, and since it is a peptide drug, it was not included in the subsequent construction of small molecule compounds.

[0076] Of these eight compounds, VUF11207 and AMD3100 (S3013) are known CXCR7 agonists, while the activation of CXCR7 by the remaining six small molecule compounds has not been reported.

[0077] Therefore, by utilizing the PRESTO-Tango drug screening platform and the GPCR small molecule drug library, this embodiment discovered six novel CXCR7 agonists, including S0135 (JMS-17-2, EC50). 50 The value is 70.7 nM), S3052 (Rupatadine, EC) 50 The value is 384.8 nM), S1151 (MK-0667, ibuprofen mesylate, EC) 50 Value 3.94 μM), S1283 (Asenapine, EC) 50 Value >100 μM), S3054 (Alverine, EC) 50 Values ​​>100 μM) and S3727 (Vilanterol, EC) 50 Value >100 μM) Figure 2 ).

[0078] Example 2 To further design novel and highly efficient CXCR7 small molecule agonists, this embodiment conducted structural analysis on eight small molecule compounds identified as having CXCR7 activation potential. By constructing a structure-activity relationship (SAR) model, these compounds were fragmented, resulting in 35 fragments from the eight active compounds. Figure 2 ).

[0079] Using these fragments, this embodiment constructed 20,437 new small molecule compounds using a random combination method. These compounds all have molecular weights below 550 Da and water solubility logS values ​​between -8 and 1. Furthermore, this embodiment analyzed the fingerprint spectra of these compounds, representing the substructure of each compound as byte information based on the FP:MACCS algorithm, and then performed cluster analysis on these compounds. Compounds with a similarity greater than 85% were grouped into the same category, ultimately classifying the 20,437 compounds into 3,262 categories (…). Figure 2 ).

[0080] From each category, this embodiment selected a representative compound to form a new small molecule compound library containing a total of 3262 compounds. Based on this, this embodiment obtained the interaction and affinity score between each compound and CXCR7 through molecular docking. This embodiment screened compounds that conformed to the pharmacophore model and finally identified 23 small molecule compounds with good activity potential through affinity scores, pharmacophore model, key amino acid sites, and hydrogen bond interactions. These compounds have different structures from known active compounds.

[0081] Example 3 From the above 23 small molecule compounds, this embodiment screened out 4 new compounds, namely cpd-2-3, cpd-1-3, cpd-1-5, or cpd-2-2. Figure 3 ).

[0082] To further validate the therapeutic effects of these four new compounds on pulmonary fibrosis in vivo, this embodiment established a bleomycin-induced pulmonary fibrosis mouse model (pulmonary fibrosis was induced by intratracheal injection of 2 mg / kg bleomycin). Fourteen days after bleomycin modeling, mice were administered cpd-2-3, cpd-1-3, cpd-1-5, or cpd-2-2 via intraperitoneal injection at a dose of 10 mg / kg / day.

[0083] This example evaluated the therapeutic effects of these compounds on pulmonary fibrosis after 7 days of drug injection. The efficacy was assessed by measuring hydroxyproline content (…). Figure 4 A) Masson staining ( Figure 4 B), Sirius Red staining ( Figure 4 C) H&E staining ( Figure 4 D) and Collagen I ( Figure 4 E) and α-SMA ( Figure 4 F) Immunofluorescence staining revealed that among these new compounds, cpd-2-3 showed the most significant therapeutic effect on pulmonary fibrosis, followed by cpd-1-3. cpd-1-5 and cpd-2-2 had no significant effect on pulmonary fibrosis. The combined in vivo and in vitro results indicate that cpd-2-3 inhibits pulmonary fibrosis by efficiently activating CXCR7.

[0084] In addition, this embodiment administered different doses of cpd-2-3 intraperitoneally to mice to investigate its therapeutic effect on pulmonary fibrosis. Three doses, 2 mg / kg, 5 mg / kg, and 10 mg / kg, were tested in this embodiment. The effect was determined by hydroxyproline content assay (…). Figure 5 A) Masson staining ( Figure 5 B), Sirius Red staining (Figure 5 C) H&E staining ( Figure 5 D) and Collagen I ( Figure 5 E) and α-SMA ( Figure 5 F) Immunofluorescence staining showed that in a bleomycin-induced pulmonary fibrosis mouse model, 2 mg / kg, 5 mg / kg, and 10 mg / kg cpd-2-3 significantly inhibited pulmonary fibrosis, mainly manifested as reduced collagen deposition, reduced lung injury, and reduced fibroblast activation. Figure 5 Therefore, at low doses (2 mg / kg), the novel CXCR7 agonist cpd-2-3 also showed good efficacy in the treatment of pulmonary fibrosis.

[0085] Previous studies have found that vascular endothelial cells regulate lung regeneration and fibrosis by interacting with adjacent epithelial and interstitial cells through paracrine secretion of the angiocrine factor. To investigate the molecular mechanism of cpd-2-3 treatment for pulmonary fibrosis, this study used magnetic bead sorting technology to isolate pulmonary vascular endothelial cells and detected changes in the expression of angiocrine factors associated with lung regeneration and fibrosis.

[0086] like Figure 6 As shown, cpd-2-3 upregulates the expression of pro-regenerative angiocrine factors (Hgf, Apln, Mmp14, Dll4, Kit, Egfl7) and inhibits the expression of pro-fibrotic angiocrine factors (Tgfb1, Pdgfb, Igfbp7, Ctgf, Adamts1, Tgfb2). These results indicate that the novel agonist cpd-2-3 promotes lung regeneration and inhibits fibrosis by efficiently activating CXCR7 in endothelial cells and reprogramming angiocrine signaling in the vascular microenvironment.

[0087] Example 4 This embodiment utilizes the PRESTO-Tango drug screening system, GPCR drug library, and high-throughput small molecule drug design platform to obtain a novel CXCR7 agonist, cpd-2-3, which can efficiently activate CXCR7. Based on cpd-2-3, this embodiment also found that compounds having Formula I can act as CXCR7 agonists, efficiently activating endothelial cell CXCR7, reprogramming the angiocrine signaling in the vascular microenvironment, thereby promoting organ regeneration and inhibiting fibrosis. (I) Where L is a (C1-C6) alkyl group; A is a saturated 3-6 membered aliphatic ring, heterocyclic ring, bridged ring, or spirocyclic ring; R is -C(=O)NHR xOr -C(=O)OR x Or substituted aryl or aromatic heterol groups; R x It can be a hydrogen atom, methyl, ethyl, propyl, isopropyl, butyl, or tert-butyl.

[0088] Furthermore, this embodiment also found that compounds having formula IV can act as CXCR7 agonists (e.g., cpd-2-3 and JMS-17-2), more efficiently activating endothelial cell CXCR7, thereby better promoting organ regeneration and inhibiting fibrosis: (IV) Wherein Rz is a substituent of at least one of ortho-substitution, meta-substitution, and para-substitution, wherein the substituent is selected from the group consisting of halogen, methoxy, methyl, ethyl, propyl, isopropyl, butyl, tert-butyl, trifluoromethyl, and trifluoromethoxy.

[0089] In some embodiments, the halogen is selected from the group consisting of fluorine, bromine, and chlorine.

[0090] In some embodiments, Rz is a halogen or methoxy group that is ortho-substituted, meta-substituted, or para-substituted.

[0091] In some embodiments, the halogen is selected from the group consisting of fluorine, bromine, and chlorine.

[0092] The synthetic route for Cpd-2-3 (Equation II) is shown below. Figure 7 Numbers 1-5 correspond to compounds 1-5.

[0093] (II) The synthesis method of cpd-2-3 is as follows: (1) Preparation method of compound 2: A solution of compound 1 (5 g, 20.24 mmol, 1 equivalent) was added to a mixture of N-benzyloxycarbonyl-3,6-dihydro-2H-pyridine-4-boronic acid pinacol ester (8.33 g, 24.28 mmol, 1.2 equivalent), dioxane (40 mL), and water (10 mL), followed by the addition of Pd(PPh3)4 (2.34 g, 2.02 mmol, 0.1 equivalent) and potassium carbonate (8.39 g, 60.71 mmol, 3 equivalent). The mixture was stirred at 80°C for 5 hours under nitrogen protection. The desired compound was detected by LCMS. The reaction mixture was poured into 30 mL of water and extracted with ethyl acetate (30 mL × 3). The organic phases were combined and extracted again with water (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (silica, petroleum ether / ethyl acetate = 4 / 1 to 3 / 1). Compound 2 (3 g, 7.82 mmol, 38.66% yield) was obtained as a yellow oil.

[0094] (2) Preparation method of compound 3: Compound 2 (1 g, 2.61 mmol, 1 equivalent) was dissolved in tetrahydrofuran (THF, 20 mL), followed by the addition of a Pd / C catalyst (2.78 g, 2.61 mmol, 10% purity, 1 equivalent). The mixture was stirred at 25°C and a hydrogen pressure of 15 Psi for 2 hours. The desired compound was detected by LCMS. The reaction mixture was filtered, and the filtrate was concentrated to give compound 3 (0.65 g, crude) as a colorless oil.

[0095] (3) Preparation method of compound 5: Compound 4 (0.4 g, 2.17 mmol, 1 equivalent) was dissolved in dimethylformamide (DMF, 8 mL), followed by the addition of cesium carbonate (Cs₂CO₃, 2.12 g, 6.51 mmol, 3 equivalent) and 1,3-dibromopropane (4.38 g, 21.72 mmol, 2.21 mL, 10 equivalent). The mixture was stirred at 25°C for 1 hour. The desired compound was detected by LCMS. The organic layers were combined and concentrated under reduced pressure to obtain a residue. The residue was poured into 10 mL of water and extracted with ethyl acetate (EtOAc, 10 mL × 3). The organic phases were combined and extracted again with water (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give compound 5 (0.65 g, crude) as a yellow oil.

[0096] (4) Preparation method of Cpd-2-3: Compound 5 (0.3 g, 983.06 μmol, 1 equivalent) and compound 3 (296.48 mg, 1.18 mmol, 1.2 equivalent) were dissolved in dimethylformamide (DMF, 3 mL), followed by the addition of cesium carbonate (Cs₂CO₃, 960.90 mg, 2.95 mmol, 3 equivalent). The mixture was stirred at 25°C for 1 hour. The desired compounds were detected by LCMS. The reaction mixture was filtered, and the filtrate was collected. The crude product was purified by preparative high-performance liquid chromatography (prep-HPLC, FA conditions) [column: Phenomenexluna C18 100*40mm*5 μm; mobile phase: [H2O(0.2% FA)-ACN]; gradient: 10%-50% B over 8.0 min] to give compound Cpd-2-3 (50 mg, formate (FA) salt, 104.74 μmol, yield 10.65%, purity 99.62%) as a white solid. LCMS (ESI+): m / z 476.2 (M+H+).

[0097] Spectrum (Cpd-2-3): 1 H NMR (400 MHz, CHLOROFORM-d) δ = 8.43 (s, 1H), 7.69 - 7.58 (m, 2H), 7.41 - 7.33 (m, 1H), 7.31 -7.23 (m, 1H), 7.19 - 7.12 (m, 2H), 6.63 - 6.57 (m, 1H), 6.36 (s, 2H), 4.29(t, J = 7.3 Hz, 2H), 3.78 (s, 6H), 3.75 (s, 3H), 3.26 (br d,J= 11.6 Hz, 2H), 2.79 - 2.69 (m, 2H), 2.52 - 2.39 (m, 1H), 2.27 (dt,J= 2.6, 11.7 Hz, 2H), 2.12- 2.03 (m, 2H), 1.96 - 1.81 (m, 4H). The synthetic route for Cpd-1-3 (Formula III) is shown below. Figure 8 .

[0098] (III) The synthesis method of Cpd-1-3 is as follows: (1) General preparation steps of compound 8B: A solution of compound 8A1 (1.25 g, 10.8 mmol, 1.1 equivalents) was added to THF (40 mL), followed by N,N'-dicyclohexylcarbodiimide (DCC) (2.03 g, 9.84 mmol, 1.99 mL, 1 equivalent) and compound 8A (2 g, 9.84 mmol, 1 equivalent). The mixture was stirred at 20°C for 12 hours. Water (50 mL) was added, and the aqueous phase was extracted with EtOAc (50 mL × 3). The organic layers were combined, dried over Na2SO4, filtered, and concentrated under reduced pressure to give compound 8B (2.5 g, crude) as a white solid.

[0099] (2) General preparation steps of compound 8: A solution of compound 8B1 (650 mg, 3.33 mmol, 1 equivalent) was added to H2O (5 mL) and dioxane (20 mL), followed by the addition of TEA (triethylamine) (1.01 g, 9.99 mmol, 1.39 mL, 3 equivalents) and compound 8B (1 g, 3.33 mmol, 1 equivalent). The mixture was stirred at 50°C for 12 hours. Water (60 mL) was added, and the aqueous phase was adjusted to pH 4 with HCl (1 M). The phase was then extracted with EtOAc (40 mL × 3). The organic layers were combined, dried over Na2SO4, filtered, and concentrated to obtain the mixture. The mixture was purified by prep-HPLC (column: Phenomenex luna C18 250*50mm*15um; mobile phase: [H2O (0.04% HCl)-ACN]; gradient: 20%-60% B over 10.0 min) to give compound 8 (0.8 g, 2.10 mmol, 63.15% yield) as a white solid.

[0100] (3) General preparation steps of compound C2: A solution of compound C1 (4 g, 16.2 mmol, 1.0 equivalent) was added to dioxane (60 mL), followed by the addition of BrettPhos Pd G3 (733 mg, 809 μmol, 0.05 equivalent), Cs₂CO₃ (10.5 g, 32.3 mmol, 2.0 equivalent), and Boc-NHNH₂ (2.57 g, 19.43 mmol, 1.2 equivalent) under a nitrogen atmosphere. The mixture was stirred at 100°C for 2 hours. Water (70 mL) was added, and the aqueous phase was extracted with EtOAc (55 mL × 3). The organic layers were combined, washed with brine (35 mL × 2), dried over Na₂SO₄, filtered, and concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 5 / 1 to 2 / 1) to give compound C2 (2.2 g, 7.37 mmol, 45.55% yield) as a yellow oil.

[0101] (4) General preparation steps of compound C3: A solution of compound C2 (2.20 g, 7.37 mmol, 1.0 equivalent) was added to AcOH (30 mL), followed by compound 2A (1.82 g, 7.37 mmol, 1.0 equivalent). The mixture was stirred at 80°C for 16 hours. The reaction mixture was concentrated under vacuum, and then water (60 mL) was added. The aqueous phase was adjusted to pH 8 with saturated NaHCO3, and then extracted with EtOAc (60 mL × 3). The organic layers were combined, dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain compound C3 (3.5 g, crude product) as a black oil.

[0102] (5) General preparation steps of compound C4: A solution of compound C3 (3.50 g, 8.53 mmol, 1 equivalent) was added to EtOH (30 mL), followed by the addition of NaBH4 (1.00 g, 26.4 mmol, 3.1 equivalent). The mixture was stirred at 0°C for 8 hours, and then NaBH4 (1.68 g, 44.41 mmol, 5.21 equivalent) was added. The mixture was stirred at 0°C for another 4 hours. The reaction mixture was quenched with saturated NH4Cl (20 mL) at 0°C, and the aqueous phase was extracted with EtOAc (50 mL × 3). The organic layers were combined, dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain the residue. The residue was purified by prep-HPLC (column: Phenomenex luna C18 250*50mm*15μm; mobile phase: [H2O (0.04% HCl)-ACN]; gradient: 20%-50% B over 10.0 min) to give compound C4 (0.7 g, 1.70 mmol, 19.90% yield) as a white solid.

[0103] (6) General preparation steps of compound C5: A solution of compound C4 (400 mg, 969 μmol, 1.0 equivalent) was added to DCM (dichloromethane) (6 mL), followed by the addition of TEA (294 mg, 2.91 mmol, 405 μL, 3.0 equivalent) and MsCl (0.34 g, 2.97 mmol, 229 μL, 3.06 equivalent). The mixture was stirred at 0°C for 1 hour under a N2 atmosphere. The mixture was quenched with H2O (10 mL), and the aqueous phase was extracted with DCM (10 mL × 3). The organic layers were combined, washed with brine (15 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain compound C5 (0.5 g, crude) as a pink oily substance.

[0104] (7) General preparation steps of compound C6: A solution of compound C5 (500 mg, 1.02 mmol, 1.0 equivalent) was added to MeOH (5 mL), followed by the addition of Pd / C (54.2 mg, 50.9 μmol, 10% purity, 0.05 equivalent) and Pd(OH)2 (71.5 mg, 50.96 μmol, 10% purity, 0.05 equivalent) under N2. The suspension was degassed under vacuum and washed several times with H2, then stirred at 50°C for 12 hours under H2 (50 psi). The reaction mixture was filtered, and the filtrate was concentrated to obtain the residue. The residue was purified by prep-HPLC (column: Phenomenexluna C18 250*50mm*15μm; mobile phase: [H2O (0.04% HCl)-ACN]; gradient: 20%-70% B over 10.0 min) to give compound C6 (0.12 g, 336.67 μmol, 33.03% yield) as a white solid.

[0105] (8) General preparation steps of compound C7: A solution of compound 8 (179 mg, 471 μmol, 2 equivalents) was added to ACN (5 mL), followed by DCC (107 mg, 518 μmol, 105 μL, 2.2 equivalents) and HOBt (70.0 mg, 518 μmol, 2.2 equivalents). The mixture was stirred at -30°C for 0.2 h under a N2 atmosphere. Then, DIEA (N,N-diisopropylethylamine) (76.15 mg, 589.17 μmol, 102.62 μL, 2.5 equivalents) and compound C6 (84 mg, 235.67 μmol, 1 equivalent) were added. The mixture was stirred at -30 to 20°C for 0.8 h. The reaction mixture was concentrated under reduced pressure to obtain a residue, then water (10 mL) was added and the aqueous phase was extracted with EtOAc (10 mL × 3). The organic layers were combined, washed with brine (15 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to obtain compound C7 (0.17 g, crude product) as a white oil.

[0106] (9) General preparation steps of Cpd-1-3: A solution of compound C7 (160 mg, 222 μmol, 1.0 equivalent) was added to DCM (3 mL), followed by TFA (trifluoroacetic acid) (126 mg, 1.11 mmol, 82.6 μL, 5.0 equivalent). The mixture was stirred at 20°C for 2.5 h. The reaction mixture was concentrated under reduced pressure to obtain a residue, which was purified by prep-HPLC (column: Phenomenex luna C18 100*40mm*5 μm; mobile phase: [H2O (0.04% HCl)-ACN]; gradient: from 25% to 45% B over 8.0 min) to give Cpd-1-3 (21.8 mg, 32.9 μmol, yield 14.81%, purity 99.08%, containing HCl) as a white solid. LCMS (ESI+): m / z 619.2 (M+H + ) Spectrum (Cpd-1-3): 1 H NMR(400 MHz, DMSO-d6, T= 273 + 80K) 8.39 (d, J = 8.0 Hz, 1H), 8.31 (br s, 3H), 7.36 - 7.30 (m, 5H), 6.61 (s, 1H), 5.05 - 4.99 (m, 1H), 4.56 - 4.52 (m, 2H), 4.25 (m, 4H), 4.01 (s,2H), 3.90 (s, 3H), 3.85 - 3.80 (m, 1H), 3.79 - 3.76 (m, 6H), 3.68 - 3.63 (m,1H), 3.32 (s, 3H), 1.83 - 1.68 (m, 2H), 1.66 - 1.58 (m, 2H), 1.55 (s, 3H),1.53 (s, 3H) The embodiments of the present invention have been described above with reference to the accompanying drawings. However, the present invention is not limited to the specific embodiments described above. The specific embodiments described above are merely illustrative and not restrictive. Those skilled in the art can make many other forms under the guidance of the present invention without departing from the spirit and scope of the claims. All of these forms are within the protection scope of the present invention.

Claims

1. Use of a small molecule compound or a pharmaceutically acceptable salt thereof for the manufacture of a CXCR7 agonist, characterized in that, The small molecule compound comprises one or more of JMS-17-2, rupatadine, ibuprofen mesylate, asenapine, alverine, and vilanterol.

2. Use according to claim 1, characterized in that, The small molecule compound is JMS-17-2, ibuprofen mesylate, or rupatadine.

3. Use of a small molecule compound or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the treatment and / or prevention of organ fibrosis, characterized in that, The small molecule compound comprises one or more of JMS-17-2, rupatadine, ibuprofen mesylate, asenapine, alverine, and vilanterol.

4. Use according to claim 3, characterized in that, The small molecule compound is JMS-17-2, ibuprofen mesylate, or rupatadine.

5. The use according to claim 3, characterized in that, The organ fibrosis comprises one or more of lung fibrosis, liver fibrosis, and kidney fibrosis.

6. Use of a small molecule compound or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the treatment and / or prevention of a pulmonary disease, characterized in that, The small molecule compound comprises one or more of JMS-17-2, rupatadine, ibuprofen mesylate, asenapine, alverine, and vilanterol.

7. Use according to claim 6, characterized in that, The lung disease comprises one or more of lung fibrosis, interstitial lung disease, chronic obstructive pulmonary disease, and lung injury.

8. Use of a small molecule compound or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the treatment and / or prevention of organ damage, characterized in that, The small molecule compound comprises one or more of JMS-17-2, rupatadine, ibuprofen mesylate, asenapine, alverine, and vilanterol.

9. Use according to claim 8, characterized in that, The organ injury comprises one or more of lung injury, liver injury, and kidney injury.

10. The use according to claim 8, characterized in that, The use comprises providing a protective effect to the organ after the organ injury.

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