CXCR7 agonist and use thereof

Abstract

A CXCR7 agonist and the use thereof. A compound, wherein the compound is a compound as represented by formula I shown below, or a stereoisomer, geometric isomer, tautomer, hydrate, solvate and pharmaceutically acceptable salt or prodrug thereof. The compound as represented by formula I has a strong activation effect on CXCR7 in vitro, and can be further used as a CXCR7 agonist in application scenarios such as the treatment and / or prevention of organ fibrosis, lung diseases and / or organ damage.

Description

CXCR7 agonists and uses thereof

[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 a CXCR7 agonist and uses thereof. 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 selective binding of receptors to specific G protein subunits, causing intracellular signals to be biased to a certain pathway among numerous downstream pathways, which is known as "signal bias". Subsequent studies have shown that this bias not only manifests in selective binding to different G protein subunits to exert completely different physiological functions, but also manifests in selective bias to one of the G protein pathway or the β-arrestin pathway independent of G proteins, ultimately leading to completely different biological effects.

[0006] That is, GPCRs have the property of "signal bias", i.e., bias in activation of G proteins and β-arrestins. CXCR7 can specifically activate the intracellular β-arrestin signaling pathway without coupling to the G protein signaling pathway. This biased β-arrestin signaling transduction property helps to design CXCR7 agonists, which 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 approved for clinical use, and there is a lack of efficient CXCR7 agonists. SUMMARY

[0007] In a first aspect, the present application provides use of a small molecule compound or a pharmaceutically acceptable salt thereof in the preparation of a CXCR7 agonist, the small molecule compound comprising 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 salt forms thereof.

[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, these 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 damaged tissues or organs, 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, a "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 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.

[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 motuporin in the manufacture of a CXCR7 agonist.

[0025] In a sixth aspect, the present application also provides a compound, which 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:

[0026] wherein L is (C1-C6)alkyl;

[0027] A is a saturated 3-6 membered aliphatic ring, heterocyclic ring, bridged ring or spiro ring;

[0028] R is -C(=O)NHR x or -C(=O)OR xor a substituted aryl or arylhetero group;

[0029] 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.

[0030] In some embodiments, the compound is a compound of Formula IV:

[0031] 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.

[0032] In some embodiments, the halogen is selected from the group consisting of fluorine, bromine, and chlorine.

[0033] In some embodiments, Rz is halogen or methoxy of ortho-substitution, meta-substitution, and para-substitution.

[0034] In some embodiments, the halogen is selected from the group consisting of fluorine, bromine, and chlorine.

[0035] In some embodiments, the compound is a compound of Formula II:

[0036] In a seventh aspect, the present application also provides a compound, which 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:

[0037] In an eighth aspect, the present application also provides use of the above-mentioned compound in the preparation of a CXCR7 agonist.

[0038] In a ninth aspect, the present application also provides use of the above-mentioned compound in the preparation of a medicament for treating and / or preventing organ fibrosis.

[0039] In some embodiments, the organ fibrosis comprises one or more of pulmonary fibrosis, liver fibrosis, and kidney fibrosis.

[0040] In some embodiments, the symptoms of the organ fibrosis comprise an increase in the level of one or more of the following molecules: alpha-smooth muscle actin (a-SMA), collagen I, hydroxyproline.

[0041] In some embodiments, the medicament is used to reduce one or more of the following symptoms: the degree of organ fibrosis, the degree of organ damage, the degree of collagen deposition, the degree of fibroblast activation.

[0042] As used herein, "alkyl" includes both branched and straight-chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms. For example, (C1-C6) alkyl is intended to include alkyl groups having 1, 2, 3, 4, 5, and 6 carbon atoms. Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, n-pentyl, s-pentyl, neopentyl, and n-hexyl. In some instances, straight- chain or branched alkyl groups have six or fewer carbon atoms in their backbone (e.g., C1-C6 for straight-chain, C3-C6 for branched-chain), and in other instances, straight-chain or branched alkyl groups have four or fewer carbon atoms. Likewise, cycloalkyl groups have three to eight carbon atoms in their ring structure, and in other instances, cycloalkyl groups have five or six carbon atoms in their ring structure. Preferred are (C1-C6) alkyl groups, especially ethyl, methyl, i-propyl, i-butyl, n-pentyl, n-hexyl, and cyclopropylmethyl.

[0043] As used herein, "substituted" means that any one or more hydrogen atoms on the designated atom is replaced with the indicated group, provided that the designated atom's normal valence is not exceeded, and that the substitution results in a stable compound.

[0044] As used herein, "aryl" refers to a monocyclic or polycyclic (e.g., having 2, 3, or 4 fused rings) aromatic hydrocarbon, such as phenyl, naphthyl, anthryl, phenanthryl. In some instances, aryl groups have 6-20 carbon atoms. As used herein, the term "aromatic heterocycle" refers to an aromatic heterocyclic ring having at least one ring member that is a heteroatom, such as an O, S, or N atom. Aromatic heterocycle groups include monocyclic and polycyclic (e.g., having 2, 3, or 4 fused rings) systems. Ringed N atoms in aromatic heterocycle groups can also be oxidized to form N-oxide moieties. Examples of aromatic heterocycle groups include pyridyl, N-oxide pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, triazinyl, furanyl, quinolinyl, isoquinolinyl, thienyl, imidazolinyl, thiazolyl, indolyl, pyrrolyl, oxazolyl, benzofuranyl, benzothienyl, benzothiazolyl, isoxazolyl, pyrazolyl, triazolyl, tetrazolyl, indazolyl, 1,2,4-thiazolyl, isothiazolyl, benzothienyl, purinyl, carbazolyl, benzimidazolinyl, indolinyl. In some instances, aromatic heterocycle groups have 1 to about 20 carbon atoms, and in other instances, about 3 to 20 carbon atoms. In some instances, aromatic heterocycle groups contain 3 to about 14, 3 to about 7, or 5 to 6 ring-forming atoms. In some instances, aromatic heterocycle groups have 1 to about 4, 1 to about 3, or 1 to 2 heteroatoms.

[0045] In a tenth aspect, the present application also provides use of the above-mentioned compound in the manufacture of a medicament for treating and / or preventing a pulmonary disease.

[0046] In some embodiments, the pulmonary disease comprises one or more of pulmonary fibrosis, interstitial lung disease, chronic obstructive pulmonary disease, and lung injury.

[0047] In some embodiments, the lung injury comprises post-infection or post-surgery lung injury.

[0048] In an eleventh aspect, the present application also provides use of the compound of Formula IV in the manufacture of a medicament for treating and / or preventing organ injury.

[0049] In some embodiments, the organ comprises one or more of lung, liver, and kidney.

[0050] In some embodiments, the organ injury comprises one or more of lung injury, liver injury, and kidney injury.

[0051] In some embodiments, the use comprises providing a protective effect to the organ after the organ injury.

[0052] In a twelfth aspect, the present application also provides use of the compound of Formula IV in the manufacture of a CXCR7 agonist,

[0053] wherein Rz is a substituent of at least one of ortho-substitution, meta-substitution, and para-substitution, the substituent being selected from the group consisting of halogen, methoxy, methyl, ethyl, propyl, isopropyl, butyl, tert-butyl, trifluoromethyl, and trifluoromethoxy.

[0054] In some embodiments, the halogen is selected from the group consisting of fluorine, bromine, and chlorine.

[0055] In some embodiments, Rz is halogen or methoxy of ortho-substitution, meta-substitution, and para-substitution.

[0056] In some embodiments, the halogen is selected from the group consisting of fluorine, bromine, and chlorine.

[0057] In a thirteenth aspect, the present application also provides use of the compound of Formula IV in the manufacture of a medicament for treating and / or preventing organ fibrosis.

[0058] In some embodiments, the organ fibrosis comprises one or more of lung fibrosis, liver fibrosis, and kidney fibrosis.

[0059] In a fourteenth aspect, the present application also provides use of the compound of Formula IV in the manufacture of a medicament for treating and / or preventing lung disease.

[0060] In some embodiments, the lung disease comprises one or more of lung fibrosis, interstitial lung disease, chronic obstructive pulmonary disease, and lung injury.

[0061] In some embodiments, the lung injury comprises post-infection or post-surgery lung injury.

[0062] In a fifteenth aspect, the present application also provides use of a compound represented by Formula IV in the preparation of a medicament for treating and / or preventing organ injury.

[0063] In some embodiments, the organ comprises one or more of a lung, a liver, and a kidney.

[0064] In some embodiments, the organ injury comprises one or more of lung injury, liver injury, and kidney injury.

[0065] In some embodiments, the use comprises providing a protective effect to the organ after the organ injury.

[0066] In some embodiments, the dose of the compound ranges from 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 dose of the compound ranges from about 1, 2, 3, 4, 5, 6, 7, 8, or 9 mg / kg to about 10, 20, 30, or 40 mg / kg; from about 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 mg / kg to about 20, 30, or 40 mg / kg; from about 20, 21, 22, 23, 24, 25, 26, 27, 28, or 29 mg / kg to about 30 or 40 mg / kg; from about 30, 31, 32, 33, 34, 35, 36, 37, 38, or 39 mg / kg to about 40 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 2.0, 3.0, or 4.0 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 3.0 or 4.0 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 4.0 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; from 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 from 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.

[0067] In a sixteenth aspect, the present application provides a compound, which is a compound represented by Formula A as shown below, or a stereoisomer, a geometric isomer, a tautomer, a hydrate, a solvate, and a pharmaceutically acceptable salt or a prodrug thereof:

[0068] wherein,

[0069] L is selected from (Ci-C6)alkyl;

[0070] n is selected from 0, 1, 2, or 3;

[0071] X and Y are each independently selected from C or N;

[0072] R is selected from -C(=0)NR x R y , or -C(=0)OR x , or substituted aryl or arhetero;

[0073] R x and R y are independently selected from a hydrogen atom, a methyl group, a trifluoromethyl group, an ethyl group, a propyl group, an isopropyl group, a butyl group, a t-butyl group, a cyclopropyl group, a cyclobutyl group, a cyclopentyl group, a cyclohexyl group, a bicyclo(l,l,l)pentyl group,

[0074] R Z is a substituent that is at least one of ortho-, meta-, or para-substitution, the substituent being selected from halogen, methoxy, methyl, ethyl, propyl, isopropyl, butyl, t-butyl, trifluoromethyl, or trifluoromethoxy.

[0075] In some embodiments, R is selected from -C(=0)NR x R y or -C(=0)OR x , R x is selected from a methyl group, a trifluoromethyl group, an ethyl group, a propyl group, an isopropyl group, a butyl group, or a t-butyl group.

[0076] In some embodiments, R y is selected from a hydrogen atom.

[0077] In some embodiments, R is selected from -C(=0)NR x R y or -C(=0)OR x , R x is selected from R Z is a halogen that is at least one of ortho-, meta-, or para-substitution.

[0078] In some embodiments, the halogen is selected from fluorine, bromine, or chlorine.

[0079] In some embodiments, R y is selected from a hydrogen atom.

[0080] In some embodiments, R Z is a halogen that is para-substitution.

[0081] In some embodiments, R is selected from -C(=0)NR xR y or -C(=O)OR x , R x is selected from ethyl, propyl, isopropyl, butyl or tert-butyl.

[0082] In some embodiments, R y is selected from a hydrogen atom.

[0083] In some embodiments, L is selected from C3 or C4 alkyl.

[0084] In some embodiments, X is N and Y is selected from C or N.

[0085] In some embodiments, n is selected from 0 or 1.

[0086] In some embodiments, the compound has a formula selected from the group consisting of:

[0087] In some embodiments, the compound is a compound of Formula B:

[0088] wherein X and Y are each independently selected from C or N;

[0089] R is selected from -C(=O)NHR x , or -C(=O)OR x , or substituted aryl or arylhetero group;

[0090] R x is selected from a hydrogen atom, methyl, ethyl, propyl, isopropyl, butyl or tert-butyl.

[0091] In some embodiments, R x is selected from ethyl, propyl, isopropyl, butyl or tert-butyl.

[0092] In some embodiments, X is N and Y is selected from C or N.

[0093] In a seventeenth aspect, the present application also provides use of the compound of the sixteenth aspect in the preparation of a CXCR7 agonist.

[0094] In an eighteenth aspect, the present application also provides use of the compound of the sixteenth aspect in the preparation of a medicament for treating and / or preventing organ fibrosis.

[0095] In some embodiments, the organ fibrosis comprises one or more of pulmonary fibrosis, liver fibrosis and kidney fibrosis.

[0096] In some embodiments, the symptom of organ fibrosis comprises an increase in the level of one or more of the following molecules: a-smooth muscle actin (a-SMA), collagen I, hydroxyproline.

[0097] In some embodiments, the medicament is for reducing one or more of the following symptoms: the degree of organ fibrosis, the degree of organ injury, the degree of collagen deposition, the degree of fibroblast activation.

[0098] In a nineteenth aspect, the present application also provides use of the compound of the sixteenth aspect in the preparation of a medicament for treating and / or preventing one or more of a pulmonary disease, a liver disease, and a kidney disease.

[0099] As used herein, a "pulmonary disease" refers to a disease that occurs in the lungs, including a disease of the lungs per se 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.

[0100] In some embodiments, the lung injury comprises post-infection or post-surgery lung injury.

[0101] As used herein, a "liver disease" refers to a disease that occurs in the liver, including a disease of the liver per se or a hepatic manifestation of a systemic disease. In some embodiments, the liver disease comprises liver fibrosis and / or liver injury.

[0102] As used herein, a "kidney disease" refers to a disease that occurs in the kidney, including a disease of the kidney per se or a renal manifestation of a systemic disease. In some embodiments, the kidney disease comprises kidney fibrosis and / or kidney injury.

[0103] In a twentieth aspect, the present application also provides use of the compound of the sixteenth aspect in the preparation of a medicament for treating and / or preventing an organ injury.

[0104] In some embodiments, the organ comprises one or more of the lung, the liver, and the kidney.

[0105] In some embodiments, the organ injury comprises one or more of lung injury, liver injury, and kidney injury.

[0106] In some embodiments, the use comprises providing a protective effect to the organ after the organ injury.

[0107] In some embodiments, the dose of the compound ranges from about 0.1 mg / kg to about 40 mg / kg, 0.1 mg / kg to about 4 mg / kg, or 0.01 mg / kg to about 1.00 mg / kg. In some embodiments, the dose of the compound ranges from about 1, 2, 3, 4, 5, 6, 7, 8, or 9 mg / kg to about 10, 20, 30, or 40 mg / kg; from about 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 mg / kg to about 20, 30, or 40 mg / kg; from about 20, 21, 22, 23, 24, 25, 26, 27, 28, or 29 mg / kg to about 30 or 40 mg / kg; from about 30, 31, 32, 33, 34, 35, 36, 37, 38, or 39 mg / kg to about 40 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 2.0, 3.0, or 4.0 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 3.0 or 4.0 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 4.0 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; from 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 from 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.

[0108] The advantageous effects of the present application compared with the prior art include at least the following aspects:

[0109] Previous studies found that CXCR7 signaling is inhibited in the vascular microenvironment of fibrotic lung tissue, leading to pro-fibrotic Angiocrine signals that stimulate perivascular fibroblasts, thereby activating fibrotic responses and suppressing lung regeneration. While activation of the CXCR7 receptor in the pulmonary vascular microenvironment reprograms Angiocrine signals in the vascular microenvironment, inhibits fibroblast activation, and stimulates alveolar stem cell viability, thereby promoting repair and regeneration of fibrotic lung tissue. However, there is still a lack of efficient CXCR7 agonists.

[0110] The present application identifies six small molecule compounds (i.e., JMS-17-2, rupatadine, ibuprofen mesylate, asenapine, alverine, and vilanterol) that can specifically activate the intracellular β-arrestin signaling pathway without coupling the G protein signaling pathway, thereby serving as novel CXCR7 agonists. Although the above six small molecule compounds are all targeted GPCRs, the known uses of the above six small molecule compounds 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 a platelet-activating factor inhibitor; ibuprofen mesylate is a selective ghrelin receptor agonist and growth hormone secretagogue (GHSR) that mimics the growth hormone (GH) stimulating effect of the endogenous hormone ghrelin; asenapine is an atypical antipsychotic drug with central nervous system inhibitory and anti-histamine and anti-serotonin activities; alverine is a synthetic papaverine derivative that is a selective smooth muscle relaxant; and vilanterol is a novel β2-receptor agonist. That is, the activation of CXCR7 by the above six small molecule compounds has not been reported. Furthermore, the above six small molecule compounds as CXCR7 agonists can be further used for treating and / or preventing organ fibrosis, lung diseases, and / or organ damage, and the like.

[0111] The synthesized compounds (for example, the compounds represented by formula I, III, IV and the compound represented by formula A) have strong activation effect on CXCR7 in vitro. In the CXCR7 activity test, the EC50 (half effective concentration) of most of the compounds (for example, the compound represented by formula IV (for example, formula II)) reaches the nanomolar level, which is even higher than that of the existing CXCR7 agonist, for example, VUF11207. The synthesized compounds can specifically activate the intracellular β-arrestin signal pathway without coupling the G protein signal pathway, thereby serving as a new type of CXCR7 agonist, efficiently activating CXCR7 in endothelial cells, reprogramming the Angiocrine signal of the vascular microenvironment, and promoting organ regeneration and inhibiting organ fibrosis. Further, as the CXCR7 agonist, the synthesized compounds can be further used for treating and / or preventing organ fibrosis, lung diseases and / or organ damage and the like application scenarios. BRIEF DESCRIPTION OF DRAWINGS

[0112] In order to more clearly illustrate the technical solutions in the embodiments of the present application or the prior art, the drawings needed to be used in the embodiments or the prior art description will be briefly introduced below. In all the drawings, similar elements or parts are generally identified by similar reference numerals. In the drawings, each element or part is not necessarily drawn according to the actual scale. Obviously, the drawings described below are some embodiments of the present application, and those skilled in the art can obtain other drawings according to these drawings without any creative labor.

[0113] Figure 1 is a result graph of discovering new CXCR7 agonists based on the PRESTO-Tango high-throughput drug screening platform;

[0114] Figure 2 is a technical roadmap of designing new small molecule agonists of CXCR7;

[0115] Figure 3 is a result graph of testing the activation potential of cpd-2-3, cpd-1-3, cpd-1-5 and cpd-2-2 on CXCR7 by using the PRESTO-Tango system (the results are represented by mean ± SEM (n = 3));

[0116] Figure 4 is a result graph of verifying the treatment effect of the four new compounds on pulmonary fibrosis in vivo;

[0117] Figure 5 is a result graph of the treatment effect of different doses of cpd-2-3 on pulmonary fibrosis;

[0118] Figure 6 is a result graph of cpd-2-3 inhibiting pulmonary fibrosis;

[0119] Figure 7 is a synthesis route of cpd-2-3;

[0120] Figure 8 is a synthetic route of cpd-1-3;

[0121] Figure 9 is a synthetic route of compound S1;

[0122] Figure 10 is a synthetic route of compound S2;

[0123] Figure 11 is a synthetic route of compound S3;

[0124] Figure 12 is a synthetic route of compound S4;

[0125] Figure 13 is a synthetic route of compound S5;

[0126] Figure 14 is a synthetic route of compound S6;

[0127] Figure 15 is a synthetic route of compound S7;

[0128] Figure 16 is a synthetic route of compound S8;

[0129] Figure 17 is a synthetic route of compound S9;

[0130] Figure 18 is a result graph of the activation effect of compound S1 on CXCR7;

[0131] Figure 19 is a result graph of the activation effect of compounds S1, S2 and S9 on CXCR7 detected by PRESTO-Tango system (the results are expressed as mean ± SEM (n = 3));

[0132] Figure 20 is a result graph of the therapeutic effect of CXCR7 agonist S1 on pulmonary fibrosis verified in vivo;

[0133] Figure 21 is a result graph of CXCR7 being specifically highly expressed in lung vascular endothelial cells;

[0134] Figure 22 is a result graph of compound S1 inhibiting pulmonary fibrosis;

[0135] Figure 23 is a result graph of the therapeutic effect of CXCR7 agonist S1 on liver fibrosis verified in vivo. DETAILED DESCRIPTION

[0136] In order to make the objectives, technical solutions and advantages of the embodiments of the present application clearer, the technical solutions in the embodiments of the present application will be described clearly and completely below with reference to the drawings in the embodiments of the present application. Obviously, the described embodiments are some but not all of the embodiments of the present application. Based on the embodiments in the present application, all other embodiments obtained by a person of ordinary skill in the art without creative work fall within the protection scope of the present application.

[0137] As used herein, "and / or", "at least one of", and "one or more of" includes any and all combinations of one or more of the associated listed items.

[0138] As used herein, the singular forms "a", "an" and "the" include plural referents unless the context clearly dictates otherwise.

[0139] It should be noted that, as used in this specification and the appended claims, the singular forms "a", "an", and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a compound" includes a mixture of two or more compounds, and the like.

[0140] As used in this specification and the appended claims, the term "approximately" typically refers to + / - 5% of the value stated, more typically + / - 4% of the value stated, more typically + / - 3% of the value stated, more typically + / - 2% of the value stated, even more typically + / - 1% of the value stated, and even more typically + / - 0.5% of the value stated.

[0141] In this specification, certain embodiments can be disclosed in a range format. It should be understood that the description in range format is merely a shorthand for describing a full range of values. The description of a range format is to be understood to include individually each possible value in the range. For example, description in the range format of 1 to 6 is to be understood to include the values of 1, 2, 3, 4, 5, and 6 individually, as well as 1 to 6 collectively. The description of a range format is to be understood to include individual values within a range, and to include individual values within a range, unless the context clearly indicates otherwise. For example, a range of 1 to 6 is to be understood to include the values of 1, 2, 3, 4, 5, and 6 individually, as well as 1 to 6 collectively. The description of a range format is to be understood to include individual values within a range, and to include individual values within a range, unless the context clearly indicates otherwise.

[0142] BRIEF DESCRIPTION OF DRAWINGS

[0143] Figure 1 : A. Technical pipeline of the PRESTO-Tango high-throughput drug screening platform to identify CXCR7 agonists. The GPCR drug library contains a total of 1132 compounds. B-C. Eight compounds were identified to be able to activate CXCR7 using the PRESTO-Tango system. Results are shown as mean ± SEM (n=3).

[0144] Figure 4: Results of hydroxyproline content assay (A), Masson staining (B), Sirius red staining (C), H&E staining (D), and Collagen I and a-SMA immunofluorescence staining (E-F) showed that cpd-2-3 had the most significant therapeutic effect on lung fibrosis, while cpd-1-3 was less effective. cpd-1-5 and cpd-2-2 had no significant effect on lung fibrosis. Results are expressed as mean ± SEM (n = 5).

[0145] Figure 5: Results of hydroxyproline content assay (A), Masson staining (B), Sirius red staining (C), H&E staining (D), and Collagen I and a-SMA immunofluorescence staining (E-F) evaluated the therapeutic effect of 2 mg / kg, 5 mg / kg, 10 mg / kg cpd-2-3 on lung fibrosis. Results are expressed as mean ± SEM (n = 5).

[0146] Figure 6: cpd-2-3 promotes lung regeneration and inhibits fibrosis by targeting endothelial cell CXCR7, reprogramming Angiocrine signals of vascular microenvironment. Lung vascular endothelial cells were isolated by magnetic bead sorting technique, and the expression changes of Angiocrine factors including Hgf, Apln, Mmp14, Dll4, Kit, Egfl7, Tgfb1, Pdgfb, Igfbp7, Ctgf, Adamts1, Tgfb2d were detected. Results are expressed as mean ± SEM (n = 3).

[0147] Figure 20: Results of hydroxyproline content assay (A), Masson staining (B), Sirius red staining (C), H&E staining (D), and Collagen I (E) and a-SMA immunofluorescence staining (F) showed that compound S1 could alleviate lung fibrosis. Results are expressed as mean ± SEM (n = 5).

[0148] Figure 21: (A-B) Single-cell transcriptome sequencing (scRNA-seq) analysis of CXCR7 expression in various cell types of lung tissue. (C) Immunofluorescence staining showed that CXCR7 was specifically expressed in lung vascular endothelial cells. VE-Cad is a marker for vascular endothelial cells.

[0149] Figure 22: Compound S1 promotes lung regeneration and inhibits fibrosis by targeting endothelial cell CXCR7, reprogramming Angiocrine signals of vascular microenvironment. Lung vascular endothelial cells were isolated by magnetic bead sorting technique, and the expression changes of Angiocrine factors including Hgf, Apln, Mmp14, Dll4, Kit, Egfl7, Tgfb1, Pdgfb, Igfbp7, Ctgf, Adamts1, Tgfb2d were detected. Results are expressed as mean ± SEM (n = 3).

[0150] Figure 23: Serum alanine aminotransferase (ALT) level test (A), Masson staining (C), Sirius red staining (B), H&E staining (D), and a-SMA immunofluorescence staining (E) results show that compound S1 can reduce liver fibrosis. The results are expressed as mean ± SEM (n = 5).

[0151] Example One

[0152] To develop new and efficient CXCR7 agonists, this embodiment uses the PRESTO-Tango high-throughput drug screening platform, combined with the GPCR small molecule drug library for screening, to find small molecule compounds with the potential to activate CXCR7. PRESTO-Tango is based on a luciferase reporter system to detect the activation of the CXCR7 coupled β-arrestin signaling pathway by small molecule drugs. It is based on the binding of ligands to specific G protein-coupled receptors (GPCRs) to activate the downstream β-arrestin signaling pathway, which is mediated by the recruitment of protease (TEV protease)-fused arrestins (β-arrestin) to the activated receptor. The protease-labeled arrestin cleaves the protease site fused to the C-terminus of the GPCR, releasing the unnatural transcription factor. This transcription factor immediately enters the nucleus, thereby bypassing additional signaling intermediates. According to the activation of the reporter gene, the degree of interaction between the target receptor and the protease-fused arrestin partner can be quantitatively determined, the 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, the intracellular β-arrestin signaling pathway can be further activated, stimulating luciferase expression (Figure 1A).

[0153] Using this system, this embodiment tested 1132 compounds in the GPCR drug library one by one, and identified 8 compounds that can activate CXCR7 (Figure 1B, Figure 1C), namely VUF11207, AMD3100, JMS-17-2, Rupatadine, MK-0667, Asenapine, Alverine, and Vilanterol. Although the EC 50 value of S9665 (Motixafortide, Motupirone) is low, since it is a polypeptide drug, it is not included in the subsequent construction of small molecule compounds.

[0154] Among these 8 compounds, VUF11207 and AMD3100 (S3013) are known CXCR7 agonists, and the remaining 6 small molecule compounds have not been reported to activate CXCR7.

[0155] Therefore, by using the PRESTO-Tango drug screening platform and the GPCR small molecule drug library, the present embodiment discovers 6 novel CXCR7 agonists, including S0135 (JMS-17-2, EC 50 value of 70.7 nM), S3052 (Rupatadine, EC 50 value of 384.8 nM), S1151 (MK-0667, EC 50 value of 3.94 μM), S1283 (Asenapine, EC 50 value of >100 μM), S3054 (Alverine, EC 50 value of >100 μM), and S3727 (Vilanterol, EC 50 value of >100 μM) (Figure 2).

[0156] Embodiment Two

[0157] In order to further design novel and efficient CXCR7 small molecule agonists, the present embodiment performs structural analysis based on the identified 8 small molecule compounds with activation potential for CXCR7. By constructing a structure-activity relationship (SAR) model, the present embodiment performs fragmentation processing on these compounds, and 35 fragments are generated from the 8 active compounds (Figure 2).

[0158] Using these fragments, the present embodiment constructs 20437 novel small molecule compounds by using a random combination method, and the molecular weight of these compounds is all below 550 Da, and the water solubility logS value is between -8 and 1. Further, the present embodiment analyzes the fingerprint of these compounds, and based on the FP:MACCS algorithm, the substructure of each compound is represented by byte information, and then clustering analysis is performed on these compounds. Compounds with a similarity greater than 85% are summarized into the same class, and finally 20437 compounds are divided into 3262 classes (Figure 2).

[0159] From each class, the present embodiment selects a representative compound to form a novel small molecule compound library, which contains 3262 compounds. On this basis, the present embodiment obtains the interaction and affinity scoring value of each compound with CXCR7 by molecular docking. The present embodiment screens compounds that meet the pharmacophore model, and finally determines 23 small molecule compounds with good activity potential by affinity scoring value, pharmacophore model, key amino acid sites, and hydrogen bond interaction, and these compounds have different structures from known active compounds.

[0160] Embodiment Three

[0161] From the above 23 small molecule compounds, four new compounds, cpd-2-3, cpd-1-3, cpd-1-5 or cpd-2-2, were screened out (Figure 3).

[0162] To further verify the therapeutic effect of the four new compounds on pulmonary fibrosis in vivo, a bleomycin-induced pulmonary fibrosis mouse model was constructed (intratracheal injection of 2 mg / kg bleomycin was used to induce pulmonary fibrosis) in this embodiment. After 14 days of bleomycin modeling, this embodiment began to administer cpd-2-3, cpd-1-3, cpd-1-5 or cpd-2-2 to mice by intraperitoneal injection, with a dose of 10 mg / kg / day.

[0163] After 7 days of drug injection, this embodiment evaluated the therapeutic effect of these compounds on pulmonary fibrosis. Through hydroxyproline content determination (Figure 4A), Masson staining (Figure 4B), Sirius red staining (Figure 4C), H&E staining (Figure 4D), and Collagen I (Figure 4E) and a-SMA (Figure 4F) immunofluorescence staining, this embodiment found that among these new compounds, 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 obvious effect on pulmonary fibrosis. The results of in vivo and in vitro studies showed that cpd-2-3 inhibited pulmonary fibrosis by efficiently activating CXCR7.

[0164] In addition, this embodiment performed intraperitoneal injection of different doses of cpd-2-3 on mice to explore its therapeutic effect on pulmonary fibrosis. This embodiment tested three doses of 2 mg / kg, 5 mg / kg and 10 mg / kg. Through hydroxyproline content determination (Figure 5A), Masson staining (Figure 5B), Sirius red staining (Figure 5C), H&E staining (Figure 5D), and Collagen I (Figure 5E) and a-SMA (Figure 5F) immunofluorescence staining, it can be seen that in the bleomycin-induced pulmonary fibrosis mouse model, 2 mg / kg, 5 mg / kg, 10 mg / kg cpd-2-3 can significantly inhibit pulmonary fibrosis, mainly manifested as reduced collagen deposition, reduced lung injury, and reduced fibroblast activation (Figure 5). Therefore, at a low dose (2 mg / kg), the new CXCR7 agonist cpd-2-3 also showed good effect on the treatment of pulmonary fibrosis.

[0165] Previous studies have found that vascular endothelial cells interact with adjacent epithelial and stromal cells through paracrine "Angiocrine" factors to regulate lung regeneration and fibrosis. In order to explore the molecular mechanism of cpd-2-3 in treating pulmonary fibrosis, the lung vascular endothelial cells were isolated by magnetic bead sorting technology in this embodiment, and the expression changes of Angiocrine factors related to lung regeneration and fibrosis were detected.

[0166] As shown in Figure 6, cpd-2-3 can up-regulate the expression of pro-regenerative Angiocrine factors (Hgf, Apln, Mmp14, Dll4, Kit, Egfl7) and inhibit the expression of pro-fibrotic Angiocrine factors (Tgfb1, Pdgfb, Igfbp7, Ctgf, Adamts1, Tgfb2). These results show that the novel agonist cpd-2-3 reprograms the Angiocrine signals in the vascular microenvironment by efficiently activating CXCR7 in endothelial cells, thereby promoting lung regeneration and inhibiting fibrosis.

[0167] Embodiment Four

[0168] In this embodiment, by using the PRESTO-Tango drug screening system, GPCR drug library and high-throughput small molecule drug design platform, a novel agonist cpd-2-3 that can efficiently activate CXCR7 is obtained. Based on cpd-2-3, this embodiment also finds that the compound having formula I can be used as a CXCR7 agonist, efficiently activates endothelial cell CXCR7, reprograms the Angiocrine signals in the vascular microenvironment, and thereby promotes organ regeneration and inhibits fibrosis:

[0169] wherein L is (C1-C6)alkyl;

[0170] A is a saturated 3-6 membered aliphatic ring, heterocyclic ring, bridged ring or spiro ring;

[0171] R is -C(=O)NHR x or -C(=O)OR x or substituted aryl or arylhetero group;

[0172] R x is a hydrogen atom, methyl, ethyl, propyl, isopropyl, butyl or tert-butyl.

[0173] Further, this embodiment also finds that the compound having formula IV can be used as a CXCR7 agonist (such as cpd-2-3 and JMS-17-2), which can more efficiently activate endothelial cell CXCR7, and thereby better promote organ regeneration and inhibit fibrosis:

[0174] wherein Rz is at least one of ortho-, meta-, para-substituted substituents selected from the group consisting of halogen, methoxy, methyl, ethyl, propyl, isopropyl, butyl, tert-butyl, trifluoromethyl and trifluoromethoxy.

[0175] In some embodiments, the halogen is selected from the group consisting of fluorine, bromine and chlorine.

[0176] In some embodiments, Rz is halogen or methoxy of ortho-, meta- and para- substitution.

[0177] In some embodiments, the halogen is selected from the group consisting of fluorine, bromine and chlorine.

[0178] The synthesis route of Cpd-2-3 (Formula II) is shown in Figure 7, and the numbers 1-5 correspond to compounds 1-5.

[0179] The synthesis method of cpd-2-3 is as follows:

[0180] (1) Preparation method of compound 2:

[0181] A solution of compound 1 (5 g, 20.24 mmol, 1 equivalent) was added N- benzyloxycarbonyl-3,6-dihydro-2H-pyridine-4-boronic acid pinacol ester (8.33 g, 24.28 mmol, 1.2 equivalents), dioxane (40 mL) and water (10 mL), then Pd(PPh3)4 (2.34 g, 2.02 mmol, 0.1 equivalent) and potassium carbonate (8.39 g, 60.71 mmol, 3 equivalents) were added. The mixture was stirred at 80°C under nitrogen protection for 5 hours. The desired compound was detected by LCMS. The reaction mixture was poured into 30 mL of water and extracted with ethyl acetate (30 mL x 3). The organic phase was combined, extracted with water (10 mL), then 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.

[0182] (2) Preparation method of compound 3:

[0183] Compound 2 (1 g, 2.61 mmol, 1 eq) was dissolved in tetrahydrofuran (THF, 20 mL), then Pd / C catalyst (2.78 g, 2.61 mmol, 10% purity, 1 eq) was added. The mixture was stirred at 25 °C under 15 Psi hydrogen pressure for 2 h. 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.

[0184] (3) Preparation method of compound 5:

[0185] Compound 4 (0.4 g, 2.17 mmol, 1 eq) was dissolved in dimethylformamide (DMF, 8 mL), then cesium carbonate (Cs2CO3, 2.12 g, 6.51 mmol, 3 eq) and 1,3-dibromopropane (4.38 g, 21.72 mmol, 2.21 mL, 10 eq) were added. The mixture was stirred at 25 °C for 1 h. The desired compound was detected by LCMS. The organic layers were combined and concentrated under reduced pressure to give a residue. The residue was poured into 10 mL of water and extracted with ethyl acetate (EtOAc, 10 mL x 3). The organic phase was combined, extracted with water (10 mL), then dried over anhydrous sodium sulfate, filtered and concentrated in vacuum to give compound 5 (0.65 g, crude) as a yellow oil.

[0186] (4) Preparation method of Cpd-2-3:

[0187] Compound 5 (0.3 g, 983.06 µmol, 1 eq) and compound 3 (296.48 mg, 1.18 mmol, 1.2 eq) were dissolved in dimethylformamide (DMF, 3 mL), then cesium carbonate (Cs2CO3, 960.90 mg, 2.95 mmol, 3 eq) was added. The mixture was stirred at 25 °C for 1 h. The desired compound was detected by LCMS. The reaction mixture was filtered, and the filtrate was collected. The crude product was purified by prep-HPLC (FA condition) [column: Phenomenex luna C18 100*40mm*5um; mobile phase: [H2O (0.2% FA) - ACN]; gradient: 10% - 50% B over 8.0 min] to give compound Cpd-2-3 (50 mg, formic acid (FA) salt, 104.74 µmol, 10.65% yield, 99.62% purity) as a white solid. LCMS (ESI+): m / z 476.2 (M+H+).

[0188] Spectrum (Cpd-2-3):

[0189] 1 H NMR (400 MHz, CHLOROFORM-d)

[0190] 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).

[0191] The synthetic route of Cpd-1-3 (Formula III) is shown in Figure 8.

[0192] The synthesis of Cpd-1-3 is as follows:

[0193] (1) General preparation procedure for compound 8B:

[0194] A solution of compound 8A1 (1.25 g, 10.8 mmol, 1.1 eq) in THF (40 mL) was added, followed by N,N'-dicyclohexylcarbodiimide (DCC) (2.03 g, 9.84 mmol, 1.99 mL, 1 eq) and compound 8A (2 g, 9.84 mmol, 1 eq), and the mixture was stirred at 20 °C for 12 h. Water (50 mL) was added, and the aqueous phase was extracted with EtOAc (50 mL x 3), the combined organic layers were dried over Na2S04, filtered, and concentrated under reduced pressure to give compound 8B (2.5 g, crude) as a white solid.

[0195] (2) General preparation procedure for compound 8:

[0196] To a solution of compound 8B1 (650 mg, 3.33 mmol, 1 eq) in H2O (5 mL) and dioxane (20 mL) was added TEA (triethylamine) (1.01 g, 9.99 mmol, 1.39 mL, 3 eq) and compound 8B (1 g, 3.33 mmol, 1 eq) and the mixture was stirred at 50 °C for 12 h. Water (60 mL) was added and the aqueous phase was adjusted to pH = 4 with HC1 (1 M) and then extracted with EtOAc (40 mL x 3), the combined organic layers were dried over Na2S04, filtered and concentrated to give a mixture. The mixture was purified by prep-HPLC (column: Phenomenex luna C18 250*50mm*15um; mobile phase: [H2O (0.04% HC1) - ACN]; gradient: 20% - 60% B in 10.0 min) to give compound 8 (0.8 g, 2.10 mmol, 63.15% yield) as a white solid.

[0197] (3) General preparation procedure for compound C2:

[0198] To a solution of compound C1 (4 g, 16.2 mmol, 1.0 eq) in dioxane (60 mL) was added BrettPhos Pd G3 (733 mg, 809 pmol, 0.05 eq) and Cs2C03 (10.5 g, 32.3 mmol, 2.0 eq) and Boc-NHNH2 (2.57 g, 19.43 mmol, 1.2 eq) under N2atmosphere. The mixture was stirred at 100 °C for 2 h. Water (70 mL) was added and the aqueous phase was extracted with EtOAc (55 mL x 3), the combined organic layers were washed with brine (35 mL x 2), dried over Na2S04, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (Si02, 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.

[0199] (4) General preparation procedure for compound C3:

[0200] To a solution of compound C2 (2.20 g, 7.37 mmol, 1.0 eq) in AcOH (30 mL) was added compound 2A (1.82 g, 7.37 mmol, 1.0 eq) and the mixture was stirred at 80 °C for 16 h. The reaction mixture was concentrated under vacuum, then water (60 mL) was added, the aqueous phase was adjusted to pH = 8 with saturated NaHCO3, then extracted with EtOAc (60 mL x 3), the combined organic layers were dried over Na2SO4, filtered and concentrated under reduced pressure to give compound C3 (3.5 g, crude) as a black oil.

[0201] (5) General preparation procedure for compound C4:

[0202] To a solution of compound C3 (3.50 g, 8.53 mmol, 1 eq) in EtOH (30 mL) was added NaBH4 (1.00 g, 26.4 mmol, 3.1 eq) and the mixture was stirred at 0 °C for 8 h, then NaBH4 (1.68 g, 44.41 mmol, 5.21 eq) was added and the mixture was stirred at 0 °C for another 4 h. The reaction mixture was quenched with saturated NH4Cl (20 mL) at 0 °C, then the aqueous phase was extracted with EtOAc (50 mL x 3), the combined organic layers were dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (column: Phenomenex luna C18 250*50mm*15um; mobile phase: [H2O (0.04% HC1) - ACN]; gradient: 20% - 50% B in 10.0 min) to give compound C4 (0.7 g, 1.70 mmol, 19.90% yield) as a white solid.

[0203] (6) General preparation procedure for compound C5:

[0204] To a solution of compound C4 (400 mg, 969 µmol, 1.0 eq) in DCM (6 mL) was added TEA (294 mg, 2.91 mmol, 405 µL, 3.0 eq) and MsCl (0.34 g, 2.97 mmol, 229 µL, 3.06 eq) and the mixture was stirred at 0 °C for 1 h under N2 atmosphere. The mixture was quenched with H2O (10 mL), then the aqueous phase was extracted with DCM (10 mL x 3), the combined organic layers were washed with brine (15 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give compound C5 (0.5 g, crude) as a pink oil.

[0205] (7) General preparation procedure for compound C6:

[0206] To a solution of compound C5 (500 mg, 1.02 mmol, 1.0 eq) in MeOH (5 mL) was added Pd / C (54.2 mg, 50.9 μmol, 10% purity, 0.05 eq) and Pd(OH)2(71.5 mg, 50.96 μmol, 10% purity, 0.05 eq) under N2. The suspension was degassed under vacuum and flushed with H2several times, then stirred at 50 °C for 12 h under H2(50 psi). The reaction mixture was filtered, and the filtrate was concentrated to give a residue. The residue was purified by prep-HPLC (column: Phenomenex luna C18 250*50mm*15μm; mobile phase: [H2O (0.04% HC1)-ACN]; gradient: 20%-70% B in 10.0 min) to give compound C6 (0.12 g, 336.67 μmol, 33.03% yield) as a white solid.

[0207] (8) General preparation procedure of compound C7:

[0208] To a solution of compound 8 (179 mg, 471 μmol, 2 eq) in ACN (5 mL) was added DCC (107 mg, 518 μmol, 105 μL, 2.2 eq) and HOBt (70.0 mg, 518 μmol, 2.2 eq) under N2atmosphere, stirred at -30 °C for 0.2 h, then added DIEA (N,N-diisopropylethylamine) (76.15 mg, 589.17 μmol, 102.62 μL, 2.5 eq) and compound C6 (84 mg, 235.67 μmol, 1 eq), the mixture was stirred at -30 to 20 °C for 0.8 h. The reaction mixture was concentrated under reduced pressure to give a residue, then added water (10 mL) and extracted the aqueous phase with EtOAc (10 mL x 3), combined the organic layers, washed with brine (15 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give compound C7 (0.17 g, crude) as a white oil.

[0209] (9) General preparation procedure of Cpd-1-3:

[0210] To a solution of compound C7 (160 mg, 222 pmol, 1.0 eq) in DCM (3 mL) was added TFA (126 mg, 1.11 mmol, 82.6 pL, 5.0 eq) and the mixture was stirred at 20 °C for 2.5 h. The reaction mixture was concentrated under reduced pressure to give a residue, which was purified by prep-HPLC (column: Phenomenex luna C18 100*40 mm*5 pm; mobile phase: [H20 (0.04% HC1) - ACN]; gradient: 25% to 45% B in 8.0 min) to give Cpd-1-3 (21.8 mg, 32.9 pmol, 14.81% yield, 99.08% purity, with HC1) as a white solid. LCMS (ESI+): m / z 619.2 (M+H + )

[0211] Spectrum (Cpd-1-3):

[0212] 1 H NMR (400 MHz, DMSO-d6, T = 273+80 K)

[0213] 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)

[0214] Example Five

[0215] The synthetic route of compound S1 is shown in Figure 9.

[0216] Step 1: Synthesis of intermediate 1-3 (methyl (2-nitrophenyl) proline ester)

[0217] A round bottom flask was charged with NaH (38 mg; 0.96 mmol) in 10 mL DMF and stirred at 0 °C for 15 min. Methyl 2-pyrrolecarboxylate (Intermediate 1-1, 100 mg; 0.80 mmol) was added and stirring was continued, after 30 min, 1-fluoro-2-nitrobenzene (Intermediate 1-2, 124 mg; 0.88 mmol) was added. The reaction mixture was then stirred at 50 °C overnight. After the reaction was complete, the reaction was quenched with water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate. The organic phase was concentrated under reduced pressure to remove the solvent to get orange red solid, which was purified by column chromatography to get 95 mg of pure Intermediate 1-3 as light yellow solid.

[0218] Step 2: Synthesis of Intermediate 1-4 (pyrrolo[l,2-a]quinoxalin-4(5H)-one)

[0219] A round bottom flask was charged with NaH (38 mg; 0.96 mmol) in 10 mL DMF and stirred at 0 °C for 15 min. Methyl 2-pyrrolecarboxylate (Intermediate 1-1, 100 mg; 0.80 mmol) was added and stirring was continued, after 30 min, 1-fluoro-2-nitrobenzene (Intermediate 1-2, 124 mg; 0.88 mmol) was added. The reaction mixture was then stirred at 50 °C overnight. After the reaction was complete, the reaction was quenched with water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate. The organic phase was concentrated under reduced pressure to remove the solvent to get orange red solid, which was purified by column chromatography to get 95 mg of pure Intermediate 1-3 as light yellow solid.

[0220] Step 3: Synthesis of Intermediate 1-6 (5-(3-chloropropyl)pyrrolo[l,2-a]quinoxalin-4(5H)-one)

[0221] A round bottom flask was charged with NaH (38 mg; 0.96 mmol) in 10 mL DMF and stirred at 0 °C for 15 min. Methyl 2-pyrrolecarboxylate (Intermediate 1-1, 100 mg; 0.80 mmol) was added and stirring was continued, after 30 min, 1-fluoro-2-nitrobenzene (Intermediate 1-2, 124 mg; 0.88 mmol) was added. The reaction mixture was then stirred at 50 °C overnight. After the reaction was complete, the reaction was quenched with water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate. The organic phase was concentrated under reduced pressure to remove the solvent to get orange red solid, which was purified by column chromatography to get 95 mg of pure Intermediate 1-3 as light yellow solid.

[0222] Step 4: Synthesis of Intermediate 1-9 (tert-butyl 4-(tert-butylcarbamoyl)piperidine-l- carboxylate)

[0223] To a solution of intermediate 1-8 (100 mg; 0.44 mmol) and HATU (251 mg; 0.66 mmol) in 10 mL of DMF, 56 μL of tert-butylamine and 219 μL of DIPEA (intermediate 1-7) were added dropwise using a syringe. The reaction was stirred at room temperature overnight. The product was monitored by lc-ms. When the reaction was complete, it was quenched with water, extracted with ethyl acetate, washed with saturated brine, and dried over anhydrous sodium sulfate. The organic phase was concentrated under reduced pressure to remove the solvent to give 90 mg of crude intermediate 1-9.

[0224] Step 5: Synthesis of intermediate 1-10 (N-(tert-butyl)piperidine-4-carboxamide)

[0225] To a solution of intermediate 1-9 (90 mg) in dichloromethane and hydrochloric acid dioxane, 122 mg of crude intermediate 1-10 was obtained after stirring at room temperature for 2 hours. The reaction was monitored by TLC. The crude product was obtained after drying.

[0226] Step 6: Synthesis of compound S1 (N-(tert-butyl)-1-(3-(4-oxopyrrolo[1,2-a]quinoxalin-5(4H)- yl)propyl)piperidine-4-carboxamide)

[0227] To a solution of intermediate 1-6 (100 mg; 0.38 mmol), intermediate 1-10 (77 mg; 0.42 mmol), potassium carbonate (105 mg; 0.76 mmol) and potassium iodide (6.3 mg; 0.038 mmol) in a round bottom flask, 10 mL of DMF was added and stirred at 80 °C for 8 hours. The reaction was diluted with water and extracted with ethyl acetate. The organic phase was concentrated under reduced pressure to remove the solvent. Purification was performed by column chromatography to give 65 mg of pure compound S1.

[0228] 1 H NMR (400 MHz, Chloroform-d) δ 7.69 (d, J = 8.1 Hz, 1H), 7.66 (t, J = 2.0 Hz, 1H), 7.47 (d, J = 8.3 Hz, 1H), 7.33 (t, J = 7.8 Hz, 1H), 7.26 - 7.18 (m, 2H), 6.66 (t, J = 3.4 Hz, 1H), 5.36 (s, 1H), 4.32 (t, J = 7.4 Hz, 2H), 3.01 (dt, J = 11.7, 3.7 Hz, 2H), 2.51 (t, J = 7.0 Hz, 2H), 2.08 - 1.92 (m, 5H), 1.84 (dd, J = 12.9, 3.5 Hz, 2H), 1.80 - 1.69 (m, 2H), 1.34 (s, 9H).

[0229] Example Six

[0230] The synthetic route of compound S2 is shown in Figure 10.

[0231] Step 1: Synthesis of compound S2 (tert-butyl 1-(3-(4-oxopyrrolo[l,2-a]quinoxalin-5(4H)- yl)propyl)piperidine-4-carboxylate)

[0232] A round bottom flask containing NaH (9 mg; 0.23 mmol) dissolved in 10 mL DMF was stirred at 0 °C for 15 min. Intermediate 1-6 (50 mg; 0.19 mmol) was added and stirring was continued, after 30 min tert-butyl 4-piperidinecarboxylate (Intermediate 2-1, 108 mg; 0.68 mmol) was added. The reaction mixture was stirred at 80 °C for 8 h. After the reaction was completed, the reaction mixture was quenched with water, extracted with ethyl acetate, washed with saturated brine, and dried over anhydrous sodium sulfate. The organic phase was concentrated under reduced pressure to remove the solvent, and purified by column chromatography to give 33 mg of pure compound S2.

[0233] 1 H NMR (400 MHz, Chloroform-d) δ 7.69 (dd, J = 8.1, 1.4 Hz, 1H), 7.65 (dd, J = 2.8, 1.5 Hz, 1H), 7.49 - 7.45 (m, 1H), 7.34 - 7.29 (m, 1H), 7.24 - 7.20 (m, 2H), 6.66 (dd, J = 3.9, 2.8 Hz, 1H), 4.32 (t, J = 7.5 Hz, 2H), 2.90 (dd, J = 10.6, 4.6 Hz, 2H), 2.48 (d, J = 7.0 Hz, 2H), 2.19 (tt, J = 11.0, 4.2 Hz, 1H), 2.06 - 1.98 (m, 2H), 1.97 - 1.91 (m, 2H), 1.91 - 1.85 (m, 2H), 1.73 (ddd, J = 13.2, 8.4, 3.5 Hz, 2H), 1.44 (s, 9H).

[0234] Example Seven

[0235] The synthetic route of compound S3 is shown in Figure 11.

[0236] Referring to Example Five, Step 3, Intermediate 3-2 can be obtained by replacing Intermediate 1-5 with Intermediate 3-1.

[0237] Referring to Example Five, Step 6, compound S3 can be obtained by replacing Intermediate 1-6 with Intermediate 3-2.

[0238] 1H NMR (400 MHz, Chloroform-d) δ 7.70 (dd, J = 8.1, 1.4 Hz, 1H), 7.66 (dd, J = 2.8, 1.5 Hz, 1H), 7.45 (dd, J = 8.6, 1.2 Hz, 1H), 7.37 (ddd, J = 8.5, 7.2, 1.4 Hz, 1H), 7.26 - 7.22 (m, 1H), 7.21 (dd, J = 3.9, 1.5 Hz, 1H), 6.67 (dd, J = 3.9, 2.8 Hz, 1H), 5.30 (s, 1H), 4.32 - 4.26 (m, 2H), 3.04 (d, J = 11.1 Hz, 2H), 2.51 (s, 2H), 2.05 (s, 1H), 2.02 (s, 1H), 1.87 (s, 2H), 1.79 (q, J = 7.4, 6.5 Hz, 4H), 1.74 - 1.69 (m, 2H), 1.34 (s, 9H).

[0239] Example Eight

[0240] The synthetic route of compound S4 is shown in Figure 12.

[0241] Referring to Example Five, Step 4, intermediate 4-2 can be obtained by using intermediate 4-1 instead of intermediate 1-7.

[0242] Referring to Example Five, Step 5, intermediate 4-3 hydrochloride salt can be obtained by using intermediate 4-2 instead of intermediate 1-9.

[0243] Referring to Example Five, Step 6, compound S4 can be obtained by using intermediate 4-3 instead of intermediate 1-10.

[0244] 1 H NMR (400 MHz, Chloroform-d) δ 7.71 (d, J = 8.0 Hz, 1H), 7.68 (dd, J = 2.9, 1.5 Hz, 1H), 7.42 (d, J = 8.4 Hz, 1H), 7.36 (t, J = 7.8 Hz, 1H), 7.24 (s, 1H), 7.21 (d, J = 3.7 Hz, 1H), 6.68 (t, J = 3.6 Hz, 1H), 6.08 (s, 1H), 4.37 (t, J = 6.9 Hz, 2H), 3.33 - 3.27 (m, 2H), 2.92 (s, 2H), 2.79 (d, J = 4.7 Hz, 3H), 2.29 - 2.10 (m, 6H), 2.02 (s, 3H).

[0245] Example Nine

[0246] The synthetic route of compound S5 is shown in Figure 13.

[0247] Following the procedure for Reference Example Five, Step 4, substituting intermediate 5-1 for intermediate 1-7, provided intermediate 5-2.

[0248] Following the procedure for Reference Example Five, Step 5, substituting intermediate 5-2 for intermediate 1-9, provided the hydrochloride salt of intermediate 5-3.

[0249] Following the procedure for Reference Example Five, Step 6, substituting intermediate 5-3 for intermediate 1-10, provided compound S5.

[0250] 1 H NMR (400 MHz, Chloroform-d) δ 7.70 (dd, J = 8.1, 1.4 Hz, 1H), 7.66 (dd, J = 2.8, 1.5 Hz, 1H), 7.48 (dd, J = 8.5, 1.3 Hz, 1H), 7.33 (ddd, J = 8.5, 7.3, 1.4 Hz, 1H), 7.26 - 7.22 (m, 1H), 7.21 (dd, J = 3.9, 1.5 Hz, 1H), 6.67 (dd, J = 3.9, 2.8 Hz, 1H), 4.33 (t, J = 7.3 Hz, 2H), 3.29 (qd, J = 7.3, 5.5 Hz, 2H), 3.01 (d, J = 11.1 Hz, 2H), 2.51 (t, J = 6.9 Hz, 2H), 2.09 - 2.01 (m, 2H), 1.97 (q, J = 7.1 Hz, 3H), 1.87 (d, J = 12.7 Hz, 2H), 1.75 (qd, J = 11.8, 3.8 Hz, 3H), 1.14 (t, J = 7.3 Hz, 3H).

[0251] Example Ten

[0252] The synthetic route for compound S6 is shown in Figure 14.

[0253] Following the procedure for Reference Example Five, Step 4, substituting intermediate 6-1 for intermediate 1-7, provided intermediate 6-2.

[0254] Following the procedure for Reference Example Five, Step 5, substituting intermediate 6-2 for intermediate 1-9, provided the hydrochloride salt of intermediate 6-3.

[0255] Following the procedure for Reference Example Five, Step 6, substituting intermediate 6-3 for intermediate 1-10, provided compound S6.

[0256] 1H NMR (400 MHz, Chloroform-d) δ 7.70 (dd, J = 8.1, 1.5 Hz, 1H), 7.67 (dd, J = 2.9, 1.5 Hz, 1H), 7.47 (dd, J = 8.5, 1.2 Hz, 1H), 7.34 (ddd, J = 8.5, 7.3, 1.4 Hz, 1H), 7.25 (dd, J = 8.1, 1.2 Hz, 1H), 7.21 (dd, J = 3.8, 1.4 Hz, 1H), 6.67 (dd, J = 3.9, 2.8 Hz, 1H), 5.40 (d, J = 8.0 Hz, 1H), 4.34 (t, J = 7.2 Hz, 2H), 4.12 - 4.02 (m, 1H), 3.07 (d, J = 11.5 Hz, 2H), 2.59 (t, J = 7.2 Hz, 2H), 2.11 (s, 1H), 2.03 (dd, J = 10.2, 4.2 Hz, 3H), 1.92 (d, J = 12.9 Hz, 2H), 1.84 - 1.76 (m, 2H), 1.14 (d, J = 6.6 Hz, 6H).

[0257] Example Eleven

[0258] The synthetic route of compound S7 is shown in Figure 15.

[0259] Following the procedure of Reference Example Five, Step 4, substituting intermediate 7-1 for intermediate 1-7, intermediate 7-2 was obtained.

[0260] Following the procedure of Reference Example Five, Step 5, substituting intermediate 7-2 for intermediate 1-9, the hydrochloride salt of intermediate 7-3 was obtained.

[0261] Following the procedure of Reference Example Five, Step 6, substituting intermediate 7-3 for intermediate 1-10, compound S7 was obtained.

[0262] 1H NMR (400 MHz, Chloroform-d) δ 7.66 (d, J = 8.1 Hz, 1H), 7.64 - 7.61 (m, 1H), 7.43 (d, J = 8.4 Hz, 1H), 7.42 - 7.37 (m, 2H), 7.33 - 7.28 (m, 1H), 7.20 (t, J = 7.7 Hz, 1H), 7.16 (d, J = 3.8 Hz, 1H), 7.10 (d, J = 8.1 Hz, 2H), 6.63 (t, J = 3.4 Hz, 1H), 6.28 (d, J = 7.0 Hz, 1H), 4.36 (d, J = 5.8 Hz, 2H), 4.27 (t, J = 7.4 Hz, 2H), 2.96 (dt, J = 11.8, 3.5 Hz, 2H), 2.45 (t, J = 6.9 Hz, 2H), 2.14 (tt, J = 11.5, 4.3 Hz, 1H), 1.98 - 1.88 (m, 4H), 1.84 (dd, J = 13.6, 3.5 Hz, 2H), 1.76 (td, J = 12.1, 3.5 Hz, 2H).

[0263] Example Twelve

[0264] The synthetic route of compound S8 is shown in Figure 16.

[0265] Following the procedure of Reference Example Five, Step 4, substituting intermediate 8-1 for intermediate 1-8, intermediate 8-2 can be obtained.

[0266] Following the procedure of Reference Example Five, Step 5, substituting intermediate 8-2 for intermediate 1-9, the hydrochloride salt of intermediate 1-10 can be obtained.

[0267] Following the procedure of Reference Example Five, Step 6, substituting intermediate 8-3 for intermediate 1-10, compound S8 can be obtained.

[0268] 1H NMR (400 MHz, Chloroform-d) δ 7.69 (dd, J = 8.2, 1.4 Hz, 1H), 7.66 (dd, J = 2.9, 1.5 Hz, 1H), 7.57 (dd, J = 8.5, 1.2 Hz, 1H), 7.35 (ddd, J = 8.6, 7.4, 1.4 Hz, 1H), 7.26 - 7.22 (m, 1H), 7.20 (dd, J = 3.9, 1.4 Hz, 1H), 6.66 (dd, J = 3.9, 2.8 Hz, 1H), 4.37 (t, J = 7.3 Hz, 2H), 3.45 - 3.38 (m, 2H), 2.64 (t, J = 6.8 Hz, 2H), 2.41 (dt, J = 12.3, 6.3 Hz, 1H), 2.11 (t, J = 11.7 Hz, 2H), 2.04 - 1.96 (m, 4H), 1.64 (dt, J = 12.2, 4.0 Hz, 4H), 1.34 (s, 9H).

[0269] Example Thirteen

[0270] The synthetic route of compound S9 is shown in Figure 17.

[0271] Step 1: Synthesis of intermediate 9-2 (tert-butyl 1-(3-chloropropyl)piperidine-4- carboxylate)

[0272] A round-bottom flask was charged with tert-butyl piperazine carboxylate (intermediate 9-1, 200 mg; 1.07 mmol), 1-bromo-3-chloropropane (intermediate 1-5, 423 mg; 2.86 mmol), potassium carbonate (967 mg; 3.21 mmol), and potassium iodide (18 mg; 0.11 mmol) in 10 mL of acetonitrile and stirred at 80 °C for 8 h. The product was monitored by lc-ms. After the reaction was complete, it was diluted with water and extracted with ethyl acetate. The organic phase was concentrated under reduced pressure to remove the solvent. This resulted in crude intermediate 9-2, 233 mg.

[0273] Step 2: Synthesis of compound S9 (tert-butyl 4-(3-(4-oxopyrrolo[l,2-a]quinoxalin-5(4H)- yl)propyl)piperazine- 1 -carboxylate)

[0274] A round-bottom flask was charged with NaH (13 mg; 0.32 mmol) in 10 mL of DMF and stirred at 0 °C for 15 min. Intermediate 1-4 (50 mg; 0.27 mmol) was added, and the stirring was continued. After 30 min, intermediate 9-2 (78 mg; 0.30 mmol) was added. The reaction mixture was stirred at 80 °C for 8 h. The reaction was then diluted with water and extracted with ethyl acetate. The organic phase was concentrated under reduced pressure to remove the solvent. Purification by column chromatography resulted in pure compound S9, 42 mg.

[0275] 1 H NMR (400 MHz, Chloroform-d) δ 7.71 (dd, J = 8.1, 1.5 Hz, 1H), 7.66 (dd, J = 2.8, 1.5 Hz, 1H), 7.44 (dd, J = 8.4, 1.3 Hz, 1H), 7.31 (ddd, J = 8.4, 7.3, 1.5 Hz, 1H), 7.26 - 7.23 (m, 1H), 7.22 (dd, J = 3.8, 1.4 Hz, 1H), 6.67 (dd, J = 3.9, 2.8 Hz, 1H), 4.37 - 4.32 (m, 2H), 3.42 (t, J = 5.1 Hz, 4H), 2.51 (t, J = 6.9 Hz, 2H), 2.41 (t, J = 5.1 Hz, 4H), 1.95 (p, J = 7.0 Hz, 3H), 1.46 (s, 9H).

[0276] Example Fourteen

[0277] This example tested the activation effect of the compound shown in Formula A on CXCR7, i.e. the activation effect of the new compound on the β-arrestin signal pathway coupled with CXCR7, to evaluate whether the new compound can act as a CXCR7 agonist, using the PRESTO-Tango system.

[0278] wherein,

[0279] L is selected from (C1-C6)alkyl;

[0280] n is selected from 0, 1, 2 or 3;

[0281] X and Y are each independently selected from C or N;

[0282] R is selected from -C(=O)NR x R y , or -C(=O)OR x , or substituted aryl or arhetero;

[0283] R x and R y are independently selected from hydrogen atom, methyl, trifluoromethyl, ethyl, propyl, isopropyl, butyl, tert-butyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, bicyclo(l,l,l)pentyl,

[0284] R Z is a substituent selected from halogen, methoxy, methyl, ethyl, propyl, isopropyl, butyl, tert-butyl, trifluoromethyl or trifluoromethoxy, which is at least one of ortho-substitution, meta-substitution or para-substitution.

[0285] Table 1

[0286] As shown in FIG. 18, FIG. 19 and Table 1, the compounds of Formula A can efficiently activate CXCR7 (e.g. <10 mM), wherein the compound S1 shows the strongest activation potential with an EC50value of 4.97 nM (FIG. 19A); the EC50values of the activation of CXCR7 by compounds S2 and S9 are 270.2 nM and 4.98 mM, respectively (FIG. 19B, FIG. 19C), that is, the compounds of Formula A can act as CXCR7 agonists and exhibit therapeutic and / or prophylactic effects on organ fibrosis, organ damage.

[0287] Example Fifteen

[0288] To verify the therapeutic effect of the new compound S1 on pulmonary fibrosis in vivo, this embodiment constructed a mouse model of pulmonary fibrosis by intratracheal injection of bleomycin (pulmonary fibrosis was induced by intratracheal injection of 2 mg / kg of bleomycin). After 14 days of bleomycin modeling, this embodiment intraperitoneally injected the mice with different doses of compound S1 (0.5 mg / kg, 1 mg / kg, 2 mg / kg, 5 mg / kg) to evaluate its therapeutic effect on pulmonary fibrosis.

[0289] After 7 consecutive days of drug injection, this embodiment evaluated the therapeutic effect of compound S1 on pulmonary fibrosis. Using hydroxyproline content determination (FIG. 20A), Masson staining (FIG. 20B), Sirius red staining (FIG. 20C), H&E staining (FIG. 20D), and Collagen I (FIG. 20E) and a-SMA immunofluorescence staining (FIG. 20F) techniques, this embodiment found that different doses of compound S1 can significantly reduce collagen deposition in lung tissue, reduce lung damage, and inhibit fibroblast activation, indicating that the CXCR7 agonist compound S1 can alleviate pulmonary fibrosis. Moreover, compound S1 can significantly alleviate pulmonary fibrosis at a low dose of 0.5 mg / kg, indicating that the compound has a fast onset of action for the treatment of pulmonary fibrosis (FIG. 20). Both in vivo and in vitro results indicate that compound S1 inhibits pulmonary fibrosis by efficiently activating CXCR7.

[0290] Example Sixteen

[0291] To further explore which cells in the lung tissue are targeted by compound S1 to exert an anti-fibrotic effect, the present embodiment used single-cell transcriptome sequencing (scRNA-seq) and immunofluorescence staining to detect the expression and distribution of CXCR7 in mouse lung tissue. As shown in FIG. 21, the receptor CXCR7 of compound S1 was mainly expressed in vascular endothelial cells in mouse lung tissue (FIG. 21A), and the expression amount in other cells was very low or not expressed (FIG. 21B). The above results show that compound S1 inhibits pulmonary fibrosis by activating CXCR7 in lung vascular endothelial cells (FIG. 21C).

[0292] Previous studies have found that vascular endothelial cells interact with adjacent epithelial and interstitial cells through paracrine "Angiocrine" factors to regulate lung regeneration and fibrosis. To explore the molecular mechanism of compound S1 in treating pulmonary fibrosis, the present embodiment used magnetic bead sorting technology to isolate lung vascular endothelial cells and detected the expression changes of Angiocrine factors related to lung regeneration and fibrosis.

[0293] As shown in FIG. 22, compound S1 can up-regulate the expression of pro-regenerative Angiocrine factors (Hgf, Apln, Mmp14, Dll4, Kit, Egfl7) and inhibit the expression of pro-fibrotic Angiocrine factors (Tgfb1, Pdgfb, Igfbp7, Ctgf, Adamts1, Tgfb2). These results show that the novel CXCR7 agonist compound S1 can reprogram the Angiocrine signals in the vascular microenvironment by efficiently activating CXCR7 in endothelial cells, thereby promoting lung regeneration and inhibiting fibrosis.

[0294] Embodiment Seventeen

[0295] The present embodiment further explores whether the new compound S1 can also inhibit liver fibrosis. A mouse model of liver fibrosis was established by intraperitoneal injection of CCl4. After inducing liver fibrosis, the present embodiment performed intraperitoneal injection of compound S1 at different doses (0.5 mg / kg, 1 mg / kg, 2 mg / kg, 5 mg / kg) to evaluate the therapeutic effect of the compound on liver fibrosis.

[0296] After 7 consecutive days of drug injection, the therapeutic effect of compound S1 on liver fibrosis was evaluated by detecting blood biochemical indicators and pathological conditions. Using serum glutamic-pyruvic transaminase (ALT) level detection (FIG. 23A), Masson staining (FIG. 23C), Sirius red staining (FIG. 23B), H&E staining (FIG. 23D), and a-SMA immunofluorescence staining (FIG. 23E) techniques, this embodiment found that different doses of compound S1 can significantly reduce serum ALT levels, reduce collagen deposition in the liver, reduce lung damage, and inhibit the activation of hepatic stellate cells. These results show that the CXCR7 agonist S1 can alleviate liver fibrosis. Moreover, compound S1 at a low dose of 0.5 mg / kg can significantly alleviate liver fibrosis. The above results show that compound S1 has a fast onset of treatment for liver fibrosis (FIG. 23).

[0297] The therapeutic results of pulmonary fibrosis and liver fibrosis show that the compound of formula A synthesized by the present application can act as a new CXCR7 agonist, efficiently activate CXCR7 in vascular endothelial cells, reprogram Angiocrine signals in the vascular microenvironment, and further promote the repair and regeneration of fibrotic organs (e.g., lungs, liver, kidneys).

[0298] The embodiments of the present application are described above in conjunction with the accompanying drawings, but the present application is not limited to the specific embodiments described above, which are merely illustrative and not limiting. Those of ordinary skill in the art can make many forms under the inspiration of the present application without departing from the purpose of the present application and the scope protected by the claims, which are all within the protection of the present application.

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, 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, 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 pulmonary 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, ibuprofen mesylate, asenapine, alverine, and vilanterol.

7. Use according to claim 6, characterized in that, The pulmonary disease comprises one or more of pulmonary 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, 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 protection to the organ after the organ injury.

11. A compound, characterized in that, 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: L is (C1-C6)alkyl; A is a saturated 3-6 membered aliphatic, heterocyclic, bridged, or spiro ring; R is -C(=O)NHR x or -C(=O)OR x or substituted aryl or heteroaryl; 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.

12. The compound of claim 11, wherein The compound is a compound of Formula IV: 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; The halogen is selected from the group consisting of fluorine, bromine, and chlorine.

13. The compound of claim 1, wherein The compound is a compound of formula II:

14. 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:

15. Use of a compound according to any one of claims 11-14 for the manufacture of a CXCR7 agonist.

16. Use of a compound according to any one of claims 11-14 for the manufacture of a medicament for the treatment and / or prevention of organ fibrosis, which optionally comprises one or more of pulmonary fibrosis, liver fibrosis, and kidney fibrosis.

17. Use of a compound according to any one of claims 11-14 for the manufacture of a medicament for the treatment and / or prevention of a pulmonary disease.

18. The use according to claim 17, characterized in that, The pulmonary disease comprises one or more of pulmonary fibrosis, interstitial lung disease, chronic obstructive pulmonary disease, and lung injury.

19. Use of a compound according to any one of claims 11-14 for the manufacture of a medicament for the treatment and / or prevention of an organ injury.

20. The use of claim 19, wherein, The organ injury comprises one or more of lung injury, liver injury, and kidney injury.

21. A compound, characterized in that, The compound is a compound represented by Formula A as shown below, or a stereoisomer, a geometric isomer, a tautomer, a hydrate, a solvate, and a pharmaceutically acceptable salt or a prodrug thereof: wherein, L is selected from (C1-C6)alkyl; n is selected from 0, 1, 2, or 3; X and Y are each independently selected from C or N; R is selected from -C(=O)NR x R y , or -C(=O)OR x , or substituted aryl or heteroaryl; R x and R y Independently selected from hydrogen atom, methyl, trifluoromethyl, ethyl, propyl, isopropyl, butyl, tert-butyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, bicyclo(1,1,1)pentyl, R Z is at least one of ortho-, meta-, or para-substituted substituent selected from halogen, methoxy, methyl, ethyl, propyl, isopropyl, butyl, t-butyl, trifluoromethyl, or trifluoromethoxy.

22. The compound of claim 21, wherein R is selected from -C(=O)NR x R y or -C(=O)OR x , R x is selected from methyl, trifluoromethyl, ethyl, propyl, isopropyl, butyl or tert-butyl.

23. The compound of claim 21, wherein R is selected from -C(=O)NR x R y or -C(=O)OR x , R x is selected from R Z is halogen which is at least one of ortho substitution, meta substitution, para substitution.

24. The compound of any one of claims 21-23, wherein, L is selected from C3 or C4 alkyl.

25. The compound of any one of claims 21-23, wherein X is N and Y is selected from C or N.

26. The compound of claim 21, wherein The compound has a formula selected from the group consisting of:

27. Use of a compound according to any one of claims 21-26 for the manufacture of a CXCR7 agonist.

28. Use of a compound according to any one of claims 21-26 for the manufacture of a medicament for the treatment and / or prevention of organ fibrosis.

29. Use of a compound of any one of claims 21-26 for the manufacture of a medicament for the treatment and / or prevention of one or more of a pulmonary disease, a liver disease, and a kidney disease.

30. Use of a compound of any one of claims 21-26 for the manufacture of a medicament for the treatment and / or prevention of organ damage.

Images