Amino acid compound and use thereof in body inflammatory diseases

Abstract

The present invention relates to an amino acid compound and a use thereof in body inflammatory diseases. Particularly, the present invention provides a compound represented by general formula (I) and a derivative thereof, and relates to a use of the drug for the treatment of inflammatory diseases. The compound represented by general formula (I) has the advantages of high efficiency, high specificity and the like for the inflammatory diseases. Formula (I) (note: a compound of general formula (I) and a pharmaceutically acceptable salt thereof)

Description

Amino acid compounds and their use in inflammatory diseases

[0001] Priority information

[0002] This application claims priority to Chinese patent application No. 202410403521.5 filed with the China Patent Office on April 3, 2024, entitled “A class of amino acid compounds and their use in inflammatory diseases of the body”, the entire contents of which are incorporated herein by reference. Technical Field

[0003] The present invention relates to the field of biological therapy, and in particular, to a class of amino acid compounds and compositions and their application in treating inflammatory diseases of the body. Background Art

[0004] In recent years, the number of patients with inflammatory diseases related to the immune system has been increasing, including psoriasis, multiple sclerosis, ankylosing spondylitis, inflammatory bowel disease, systemic lupus erythematosus, and allergic diseases such as asthma. Numerous researchers have devoted their efforts to synthesizing and identifying small molecule drugs that can effectively treat these diseases. However, few small molecules have been successfully developed as clinically effective therapies, making the treatment of inflammatory diseases a pressing challenge. The study of inflammatory diseases is inseparable from the understanding of the immune system. Given the rapid activation and proliferation of immune cells upon encountering foreign or self-antigens, the investigation of their cellular metabolism has become a major research focus. Some researchers have demonstrated promising therapeutic effects in mouse tumor models using inhibitors of the glutamine transporter (SLC1A5 / ASCT2); however, these small molecule compounds have yet to become clinically effective in treating human tumors. Furthermore, the inhibitory effects of these existing compounds on cancer cell proliferation and their ability to kill cancer cells contradict the goal of treating inflammatory diseases.

[0005] Currently, no researchers have developed highly active, highly selective, and low-toxic compounds and their compositions as inhibitors of glutamine transporters to study their functions and therapeutic effects in inflammatory diseases. Summary of the Invention

[0006] In one aspect of the present invention, the present invention provides a compound, which is a compound represented by general formula (I) or a stereoisomer or a pharmaceutically acceptable salt of a compound represented by formula (I). In another aspect of the present invention, according to an embodiment of the present invention, the drug is used to treat inflammatory diseases.

[0007] Formula (I),

[0008] in:

[0009] n is 1, 2, or 3;

[0010] R 1 For hydrogen, C 1-18 Alkylene, C 1-18 heteroalkylene, substituted or unsubstituted arylene, substituted or unsubstituted heteroarylene;

[0011] R 2 For hydrogen, C 1-18 Alkylene, C 1-18 heteroalkylene, substituted or unsubstituted arylene, substituted or unsubstituted heteroarylene and -C(O)R 4 The substituents are composed of 4 C 1-18 Alkylene, C 1-18 heteroalkylene, substituted or unsubstituted arylene, substituted or unsubstituted heteroarylene;

[0012] R 3 For hydrogen, C 1-18 Alkylene, C 1-18 Heteroalkylene, substituted or unsubstituted arylene, substituted or unsubstituted heteroarylene, OH, NH2, SH, N3, CN, COOH, SO3H, PO3H, alkyl or aryl carbonate, alkyl or aryl sulfonate, alkyl or aryl phosphate;

[0013] L1 is selected from: single bond, C1-C6 alkylene, C1-C6 heteroalkylene, C 3-18 Cycloalkylene, C 3-18 Heterocyclylene, N(R 5 )、O、S、C(O)、N(R 5 )C(O), C(O)N(R 5 ), C(O)O, OC(O), SO, SO2, PO3, N(R 5 )SO2 or SO2N(R 5 ). Among them, R 5 Selected from: hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, heterocycloalkyl, heterocycloalkylalkyl;

[0014] Y1 is selected from: substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted alkenyl or substituted or unsubstituted alkynyl;

[0015] L2 is selected from: single bond, C1-C6 alkyl, C1-C6 heteroalkyl, C3-C6 cycloalkyl, heterocyclic group, C2-C 24 Alkenyl, partially substituted or fully substituted C2-C 24 Alkenyl, substituted or unsubstituted alkynyl, N(R 5 )、O、S、C(O)、C(O)N(R 5 )、N(R5 )C(O), C(O)O, OC(O), SO, SO2 and N(R 5 )SO2. R 5 Selected from: hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, heterocycloalkyl, heterocycloalkylalkyl;

[0016] Y2 is selected from substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted alkenyl or substituted or unsubstituted alkynyl.

[0017] According to an embodiment of the present invention, the compound represented by formula (I) has at least one of the following structures:

[0018] Demonstration compound skeleton:

[0019] Optionally, the compound comprises a compound selected from the group consisting of the following compounds (Table 1):

[0020] Table 1:

[0021] In another aspect of the present invention, the present invention provides a compound, which is a compound represented by formula (I) or a stereoisomer or a pharmaceutically acceptable salt of the compound represented by formula (I):

[0022] in:

[0023] n is 1, 2, or 3;

[0024] R 1 For hydrogen, C 1-18 Alkylene, C 1-18 heteroalkylene, substituted or unsubstituted arylene, substituted or unsubstituted heteroarylene;

[0025] R 2 For hydrogen, C 1-18 Alkylene, C 1-18 heteroalkylene, substituted or unsubstituted arylene, substituted or unsubstituted heteroarylene and -C(O)R 4 The substituents are composed of 4 C 1-18 Alkylene, C 1-18 heteroalkylene, substituted or unsubstituted arylene, substituted or unsubstituted heteroarylene;

[0026] R 3 For hydrogen, C 1-18 Alkylene, C1-18 Heteroalkylene, substituted or unsubstituted arylene, substituted or unsubstituted heteroarylene, OH, NH2, SH, N3, CN, COOH, SO3H, PO3H, alkyl or aryl carbonate, alkyl or aryl sulfonate, alkyl or aryl phosphate;

[0027] L1 is selected from: single bond, C1-C6 alkylene, C1-C6 heteroalkylene, C 3-18 Cycloalkylene, C 3-18 Heterocyclylene, N(R 5 )、O、S、C(O)、N(R 5 )C(O), C(O)N(R 5 ), C(O)O, OC(O), SO, SO2, PO3, N(R 5 )SO2 or SO2N(R 5 ), where R 5 Selected from: hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, heterocycloalkyl, heterocycloalkylalkyl;

[0028] Y1 is selected from: substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted alkenyl or substituted or unsubstituted alkynyl;

[0029] L2 is selected from: single bond, C1-C6 alkyl, C1-C6 heteroalkyl, C3-C6 cycloalkyl, heterocyclic group, C2-C 24 Alkenyl, partially substituted or fully substituted C2-C 24 Alkenyl, substituted or unsubstituted alkynyl, N(R 5 )、O、S、C(O)、C(O)N(R 5 )、N(R 5 )C(O), C(O)O, OC(O), SO, SO2 and N(R 5 )SO2,R 5 Selected from: hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, heterocycloalkyl, heterocycloalkylalkyl;

[0030] Y2 is selected from: substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted alkenyl or substituted or unsubstituted alkynyl;

[0031] The compound represented by formula (I) has at least one of the following structures:

[0032] Demonstration compound skeleton:

[0033] According to an embodiment of the present invention, the compound includes compounds selected from the group consisting of compounds shown in Table 1.

[0034] Use of pharmaceutical composition in preparing medicine

[0035] In another aspect of the present invention, the present invention also provides the use of a pharmaceutical composition in the preparation of a drug. According to an embodiment of the present invention, the pharmaceutical composition is used to treat inflammatory diseases, and the pharmaceutical composition includes the aforementioned compound. According to an embodiment of the present invention, the inventors have discovered for the first time that the pharmaceutical composition can be used to treat inflammatory diseases. Therefore, the invention also discovered for the first time that the pharmaceutical composition can be used to treat inflammatory diseases, and has the advantages of high efficacy, high specificity, and low toxicity.

[0036] According to an embodiment of the present invention, the above-mentioned use further includes at least one of the following technical features:

[0037] According to an embodiment of the present invention, the pharmaceutical composition further comprises a pharmaceutical excipient.

[0038] According to an embodiment of the present invention, the content of the compound is 0.01-500 mg / kg. It should be noted that the content here refers to the amount of drug required to be given to each kg of mouse, which can be converted into the content required for children or adult patients based on conventional conversion. The inventors found that if the dose is too high, it will have obvious side effects on the mouse intestine, which can be lethal in severe cases. If the dose is too low, the therapeutic effect is poor.

[0039] According to an embodiment of the present invention, the inflammatory disease is selected from autoimmune diseases and allergic diseases such as asthma.

[0040] According to an embodiment of the present invention, the inflammatory disease is caused by the differentiation of at least one of T cells, dendritic cells, eosinophils, neutrophils, macrophages, innate lymphocytes, and monocytes. The inventors have discovered that the compound combination can inhibit the differentiation of cells such as T cells, dendritic cells, eosinophils, neutrophils, macrophages, innate lymphocytes, and monocytes.

[0041] According to an embodiment of the present invention, the autoimmune disease is an immune response caused by self-antigens.

[0042] According to an embodiment of the present invention, the asthma is induced by allergens.

[0043] According to an embodiment of the present invention, the compound can be used alone or in combination with other compounds to treat inflammatory diseases.

[0044] In one aspect, the present invention provides a method for treating an inflammatory disease. According to an embodiment of the present invention, the method comprises administering a pharmaceutically effective amount of the aforementioned compound to a patient with the inflammatory disease. According to the embodiments of the present invention, the inventors have discovered for the first time that the compound can be used to treat an inflammatory disease. Therefore, the compound is also the first to be discovered by the inventors for use in treating an inflammatory disease and has advantages such as high efficacy, high specificity, and low toxicity.

[0045] According to an embodiment of the present invention, the above method further includes at least one of the following technical features:

[0046] According to an embodiment of the present invention, the content (effective amount) of the compound is 0.01-500 mg / kg. It should be noted that the content here refers to the amount of drug that needs to be given to each kg of mouse, which can be converted into the content required by children or adult patients according to conventional conversion. The inventors found that if the dose is too high, it has obvious side effects on the mouse intestine, which can be lethal in severe cases. If the dose is too low, the therapeutic effect is poor.

[0047] According to an embodiment of the present invention, the inflammatory disease is selected from autoimmune diseases and asthma.

[0048] According to an embodiment of the present invention, the inflammatory disease is caused by the differentiation of at least one of T cells, dendritic cells, eosinophils, neutrophils, macrophages, innate lymphocytes, and monocytes. The inventors have discovered that the compound combination can inhibit the differentiation of cells such as T cells, dendritic cells, eosinophils, neutrophils, macrophages, innate lymphocytes, and monocytes.

[0049] According to an embodiment of the present invention, the autoimmune disease is an immune response caused by self-antigens.

[0050] According to an embodiment of the present invention, the asthma is induced by allergens.

[0051] In another aspect of the present invention, the present invention provides the use of the aforementioned compounds in treating inflammatory diseases. According to the embodiments of the present invention, the inventors have discovered for the first time that the compounds can be used to treat inflammatory diseases. Therefore, the use of the compounds in treating inflammatory diseases is also the first discovery by the inventors, and they have the advantages of high efficacy, high specificity, and low toxicity.

[0052] According to an embodiment of the present invention, the above-mentioned use further includes at least one of the following technical features:

[0053] According to an embodiment of the present invention, the content (effective amount) of the compound is 0.01-500 mg / kg. It should be noted that the content here refers to the amount of drug that needs to be given to each kg of mouse, which can be converted into the content required by children or adult patients according to conventional conversion. The inventors found that if the dose is too high, it has obvious side effects on the mouse intestine, which can be lethal in severe cases. If the dose is too low, the therapeutic effect is poor.

[0054] According to an embodiment of the present invention, the inflammatory disease is selected from autoimmune diseases and asthma.

[0055] According to an embodiment of the present invention, the inflammatory disease is caused by the differentiation of at least one of T cells, dendritic cells, eosinophils, neutrophils, macrophages, innate lymphocytes, and monocytes. The inventors have discovered that the compound combination can inhibit the differentiation of cells such as T cells, dendritic cells, eosinophils, neutrophils, macrophages, innate lymphocytes, and monocytes.

[0056] According to an embodiment of the present invention, the autoimmune disease is an immune response caused by self-antigens.

[0057] According to an embodiment of the present invention, the asthma is induced by allergens. BRIEF DESCRIPTION OF THE DRAWINGS

[0058] FIG1 is a diagram showing the results of an in vitro cell experiment on the amino acid compounds in Test Example 1.

[0059] FIG2 is a diagram showing the results of a model experiment in which the amino acid compounds in Test Example 2 were applied to multiple sclerosis mice. DETAILED DESCRIPTION

[0060] Definitions and General Terms

[0061] Certain embodiments of the present invention are now described in detail, examples of which are illustrated by the accompanying structural formulas and chemical formulae. The present invention is intended to encompass all substitutions, modifications, and equivalent technical solutions, which are all included within the scope of the invention as defined in the claims. Those skilled in the art will recognize that many methods and materials similar or equivalent to those described herein can be used to practice the present invention. The present invention is in no way limited to the methods and materials described herein. In the event that one or more of the combined documents, patents, and similar materials differ from or contradict the present application (including but not limited to defined terms, term applications, described technologies, etc.), the present application shall prevail.

[0062] It will be further appreciated that certain features of the invention, which, for clarity, are described in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the invention, which, for brevity, are described in the context of a single embodiment, may also be provided separately or in any suitable subcombination.

[0063] Unless otherwise specified, all technical terms used in the present invention have the same meaning as commonly understood by those skilled in the art to which the present invention belongs. All patents and publications related to the present invention are incorporated herein by reference in their entirety.

[0064] Unless otherwise indicated, the following definitions used herein shall apply. For purposes of the present invention, chemical elements are defined in accordance with the Periodic Table of the Elements, CAS version, and Handbook of Chemistry and Physics, 75th edition, 1994. In addition, general principles of organic chemistry may be found in "Organic Chemistry," Thomas Sorrell, University Science Books, Sausalito: 1999, and "March's Advanced Organic Chemistry" by Michael B. Smith and Jerry March, John Wiley & Sons, New York: 2007, the entire contents of which are incorporated herein by reference.

[0065] Unless otherwise specified or clearly contradicted by context, the articles "a," "an," and "the" as used herein are intended to include "at least one" or "one or more." Thus, as used herein, these articles refer to one or more than one (i.e., at least one) of the objects. For example, "a component" refers to one or more components, i.e., more than one component may be contemplated for use or use in implementing the described embodiments.

[0066] As used herein, the term "subject" refers to an animal. Typically, the animal is a mammal. A subject also refers, for example, to primates (e.g., humans, male or female), cattle, sheep, goats, horses, dogs, cats, rabbits, rats, mice, fish, birds, and the like. In certain embodiments, the subject is a primate. In other embodiments, the subject is a human.

[0067] The term "patient" used in the present invention refers to humans (including adults and children) or other animals. In some embodiments, "patient" refers to humans.

[0068] The term "comprising" is an open expression, that is, including the contents specified in the present invention, but not excluding other contents.

[0069] The stereoisomers, tautomers, nitrogen oxides, solvates, metabolites, pharmaceutically acceptable salts or prodrugs thereof of the compound of formula (I) of the present invention are all within the scope of protection of the present invention.

[0070] The compounds of the present invention can be purchased commercially, prepared according to known methods, or synthesized according to routes designed using chemical synthesis theory.

[0071] "Stereoisomers" are compounds that have identical chemical constitutions but differ in the way the atoms or groups are arranged in space. Stereoisomers include enantiomers, diastereomers, conformers (rotamers), geometric (cis / trans) isomers, atropisomers, and the like.

[0072] "Chiral" refers to a molecule that is non-superimposable on its mirror image; "achiral" refers to a molecule that is superimposable on its mirror image.

[0073] "Enantiomers" refer to two non-superimposable isomers of a compound that are mirror images of each other.

[0074] "Diastereoisomers" refers to stereoisomers that have two or more chiral neutral groups whose molecules are not mirror images of each other. Diastereoisomers have different physical properties, such as melting points, boiling points, spectral properties, and reactivities. Diastereomeric mixtures can be separated by high-resolution analytical procedures such as electrophoresis and chromatography, for example, HPLC.

[0075] Stereochemical definitions and conventions used herein generally follow SP Parker, Ed., McGraw-Hill Dictionary of Chemical Terms (1984) McGraw-Hill Book Company, New York; and Eliel, E. and Wilen, S., "Stereochemistry of Organic Compounds", John Wiley & Sons, Inc., New York, 1994.

[0076] Many organic compounds exist in optically active forms, meaning they have the ability to rotate the plane of plane-polarized light. When describing an optically active compound, the prefixes D and L or R and S are used to indicate the absolute configuration of the molecule about its chiral center(s). The prefixes d and l or (+) and (-) are the symbols used to designate the rotation of plane-polarized light caused by the compound, where (-) or l indicates that the compound is left-handed. A compound prefixed with (+) or d is right-handed. A specific stereoisomer is an enantiomer, and a mixture of such isomers is called an enantiomeric mixture. A 50:50 mixture of enantiomers is called a racemic mixture or racemate, which can occur when there is no stereoselectivity or stereospecificity in a chemical reaction or process.

[0077] Any asymmetric atom (e.g., carbon, etc.) of the compounds disclosed herein can exist in a racemic or enantiomerically enriched form, such as in the (R)-, (S)-, or (R,S)-configuration. In certain embodiments, each asymmetric atom has at least 50% enantiomeric excess, at least 60% enantiomeric excess, at least 70% enantiomeric excess, at least 80% enantiomeric excess, at least 90% enantiomeric excess, at least 95% enantiomeric excess, or at least 99% enantiomeric excess in terms of the (R)- or (S)-configuration.

[0078] Depending on the choice of starting materials and process, the compounds of the present invention may exist as one of the possible isomers or a mixture thereof, such as a racemate or a mixture of diastereoisomers (depending on the number of asymmetric carbon atoms). Optically active (R)- or (S)-isomers can be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques. If the compound contains a double bond, the substituents may be in the E or Z configuration; if the compound contains a disubstituted cycloalkyl group, the cycloalkyl substituents may be in the cis or trans configuration.

[0079] Any resulting mixture of stereoisomers can be separated into the pure or substantially pure geometric isomers, enantiomers, and diastereomers on the basis of the differences in the constituent physicochemical properties, for example, by chromatography and / or fractional crystallization.

[0080] Any racemate of the resulting final product or intermediate can be separated into its optical antipodes by methods familiar to those skilled in the art using known methods, such as by separating the diastereomeric salts obtained. The racemic products can also be separated by chiral chromatography, such as high performance liquid chromatography (HPLC) using a chiral adsorbent. In particular, enantiomers can be prepared by asymmetric synthesis, for example, see Jacques, et al., Enantiomers, Racemates and Resolutions (Wiley Interscience, New York, 1981); Principles of Asymmetric Synthesis (2 nd Ed.Robert E.Gawley, Jeffrey Aubé, Elsevier, Oxford, UK, 2012); Eliel, ELStereochemistry of Carbon Compounds (McGraw-Hill, NY, 1962); Wilen, SHTables of Resolving Agents and Optical Resolutions p.268 (ELEliel, Ed., Univ. of Notre Dame Press, Notre Dame, IN 1972); Chiral Separation Techniques: A Practical Approach (Subramanian, G.Ed., Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim, Germany, 2007).

[0081] The term "tautomer" or "tautomeric form" refers to structural isomers with different energies that can be converted into each other through a low energy barrier. If tautomerism is possible (such as in solution), a chemical equilibrium of the tautomers can be reached. For example, proton tautomers (also known as prototropic tautomers) include interconversions performed by proton migration, such as keto-enol isomerization and imine-enamine isomerization. Valence tautomers include interconversions performed by the reorganization of some bonding electrons. A specific example of keto-enol tautomerism is the interconversion of pentane-2,4-dione and 4-hydroxypent-3-ene-2-one tautomers. Another example of tautomerism is phenol-keto tautomerism. A specific example of phenol-keto tautomerism is the interconversion of pyridine-4-ol and pyridine-4(1H)-one tautomers. Unless otherwise indicated, all tautomeric forms of the compounds of the invention are within the scope of the invention.

[0082] The term "prodrug" as used in the present invention refers to a compound that is converted into a compound represented by formula (I) in vivo. Such conversion is affected by the hydrolysis of the prodrug in the blood or by enzyme conversion to the parent structure in the blood or tissue. The prodrug compound of the present invention can be an ester. In the existing invention, esters that can be used as prodrugs include phenyl esters, aliphatic (C 1-24) esters, acyloxymethyl esters, carbonates, carbamates, and amino acid esters. For example, a compound of the present invention containing a hydroxyl group can be acylated to produce a prodrug form of the compound. Other prodrug forms include phosphate esters, such as these phosphate ester compounds, which are obtained by phosphorylating a hydroxyl group on the parent compound. For a complete discussion of prodrugs, see T. Higuchi and V. Stella, Pro-drugs as Novel Delivery Systems, Vol. 14 of the ACSSymposium Series, Edward B. Roche, ed., Bioreversible Carriers in Drug Design, American Pharmaceutical Association and Pergamon Press, 1987, J. Rautio et al., Prodrugs: Design and Clinical Applications, Nature Review Drug Discovery, 2008, 7, 255-270, and SJ Hecker et al., Prodrugs of Phosphates and Phosphonates, Journal of Medicinal Chemistry, 2008, 51, 2328-2345.

[0083] "Metabolite" refers to a product resulting from the in vivo metabolism of a specific compound or salt thereof. Metabolites of a compound can be identified using techniques known in the art, and their activity can be characterized using assays such as those described herein. Such products can be obtained by administering the compound through oxidation, reduction, hydrolysis, amidation, deamidation, esterification, defatting, enzymatic cleavage, and the like. Accordingly, the present invention encompasses metabolites of the compound, including metabolites produced by contacting a compound of the invention with a mammal for a sufficient period of time.

[0084] As used herein, "pharmaceutically acceptable salts" refer to organic and inorganic salts of the compounds of the present invention. Pharmaceutically acceptable salts are well known in the art, as described in S.M. Berge et al., "Describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 1977, 66: 1-19." Pharmaceutically acceptable salts formed from non-toxic acids include, but are not limited to, inorganic acid salts formed by reaction with amino groups, such as hydrochlorides, hydrobromides, phosphates, sulfates, and perchlorates, and organic acid salts such as acetates, oxalates, maleates, tartrates, citrates, succinates, and malonates, or salts obtained by other methods described in the literature, such as ion exchange. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, cyclopentylpropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, picrate, pivalate, propionate, stearate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, and the like. Salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium and N + (C 1-4 The present invention also contemplates quaternary ammonium salts formed by any compound containing a N group. Water-soluble or oil-soluble or dispersible products can be obtained by quaternization. Alkali metal or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Pharmaceutically acceptable salts further include appropriate, non-toxic ammonium, quaternary ammonium salts and amine cations formed by counter ions, such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, C 1-8 Sulfonates and aromatic sulfonates.

[0085] As used herein, a "solvate" refers to an association formed between one or more solvent molecules and a compound of the present invention. Solvents that form solvates include, but are not limited to, water, isopropanol, ethanol, methanol, dimethyl sulfoxide, ethyl acetate, acetic acid, and aminoethanol. The term "hydrate" refers to an association formed when the solvent molecule is water.

[0086] As used herein, the term "treating" any disease or condition, in some embodiments, refers to ameliorating the disease or condition (i.e., slowing or arresting or alleviating the development of the disease or at least one clinical symptom thereof). In other embodiments, "treating" refers to alleviating or improving at least one physical parameter, including physical parameters that may not be perceived by the patient. In other embodiments, "treating" refers to regulating the disease or condition physically (e.g., stabilizing a perceptible symptom) or physiologically (e.g., stabilizing a physical parameter), or both. In other embodiments, "treating" refers to preventing or delaying the onset, occurrence, or worsening of a disease or condition.

[0087] As used herein, "inflammatory disease" refers to any disease, disorder, or condition characterized by excessive inflammatory symptoms, host tissue damage, or loss of tissue function resulting from an excessive or uncontrolled inflammatory response. "Inflammatory disease" also refers to a pathological state mediated by leukocyte influx and / or neutrophil chemotaxis.

[0088] As used herein, "inflammation" refers to a local protective response caused by tissue damage or destruction, which serves to destroy, dilute, or isolate (isolate) harmful substances and damaged tissue. Inflammation is significantly associated with leukocyte influx and / or neutrophil chemotaxis. Inflammation can result from infection with pathogenic organisms and viruses, as well as non-infectious causes such as trauma or reperfusion following myocardial infarction or stroke, immune responses to foreign antigens, and autoimmune responses. Therefore, inflammatory diseases that can be treated with the compounds disclosed herein include diseases associated with specific defense system responses as well as non-specific defense system responses.

[0089] The present invention provides a pharmaceutical composition comprising a compound disclosed in the present invention, or a compound listed in the examples; and a pharmaceutically acceptable adjuvant, excipient, carrier, solvent, or a combination thereof.

[0090] It should also be recognized that certain compounds of the present invention may be used therapeutically in free form or, if appropriate, in the form of pharmaceutically acceptable derivatives thereof. Some non-limiting embodiments of pharmaceutically acceptable derivatives include pharmaceutically acceptable prodrugs, salts, esters, salts of such esters, or any other adducts or derivatives that, when administered to a patient in need thereof, directly or indirectly provide a compound of the present invention or a metabolite or residue thereof.

[0091] The pharmaceutical composition disclosed in the present invention can be prepared and packaged as bulk form, wherein a safe and effective amount of the compound shown in formula (I) can be extracted and then administered to the patient in the form of a powder or syrup. Alternatively, the pharmaceutical composition disclosed in the present invention can be prepared and packaged as a unit dosage form, wherein each physically discrete unit contains a safe and effective amount of the compound shown in formula (I). When prepared in unit dosage form, the pharmaceutical composition disclosed in the present invention can generally contain, for example, 0.5 mg to 1 g or 1 mg to 700 mg or 5 mg to 100 mg of the compound disclosed in the present invention.

[0092] As used herein, "pharmaceutically acceptable excipient" means a pharmaceutically acceptable material, mixture, or vehicle that contributes to the consistency of a dosage form or pharmaceutical composition. Each excipient, when combined, must be compatible with the other ingredients of the pharmaceutical composition to avoid interactions that could significantly reduce the efficacy of the disclosed compounds upon administration to a patient and interactions that could render the pharmaceutical composition unpharmaceutically acceptable. Furthermore, each excipient must be pharmaceutically acceptable, e.g., possess a sufficiently high degree of purity.

[0093] Suitable pharmaceutically acceptable excipients will vary depending on the specific dosage form selected. In addition, pharmaceutically acceptable excipients may be selected based on their specific function in the composition. For example, certain pharmaceutically acceptable excipients may be selected to facilitate the production of a uniform dosage form. Certain pharmaceutically acceptable excipients may be selected to facilitate the production of a stable dosage form. Certain pharmaceutically acceptable excipients may be selected to facilitate the transport or transportation of the disclosed compounds from one organ or part of the body to another organ or part of the body when administered to a patient. Certain pharmaceutically acceptable excipients may be selected to enhance patient compliance.

[0094] Suitable pharmaceutically acceptable excipients include the following types of excipients: diluents, fillers, binders, disintegrants, lubricants, glidants, granulating agents, coating agents, wetting agents, solvents, cosolvents, suspending agents, emulsifiers, sweeteners, flavoring agents, taste masking agents, coloring agents, anti-caking agents, humectants, chelating agents, plasticizers, viscosity increasing agents, antioxidants, preservatives, stabilizers, surfactants and buffers. It will be appreciated by those skilled in the art that certain pharmaceutically acceptable excipients may provide more than one function and may provide alternative functions, depending on how much of the excipient is present in the formulation and which other excipients are present in the formulation.

[0095] The skilled person has the knowledge and skill to master this area, so that they can select the suitable pharmaceutically acceptable excipient of appropriate amount for the present invention.In addition, there are a large number of resources available to the skilled person, and they describe pharmaceutically acceptable excipients, and are used to select suitable pharmaceutically acceptable excipients.Examples include Remington's Pharmaceutical Sciences (Mack Publishing Company), The Handbook of Pharmaceutical Additives (Gower Publishing Limited), and The Handbook of Pharmaceutical Excipients (the American Pharmaceutical Association and the Pharmaceutical Press).

[0096] Various carriers for configuring pharmaceutically acceptable compositions and known techniques for their preparation are disclosed in Remington: The Science and Practice of Pharmacy, 21st edition, 2005, ed. D. B. Troy, Lippincott Williams & Wilkins, Philadelphia, and Encyclopedia of Pharmaceutical Technology, eds. J. Swarbrick and J. C. Boylan, 1988-1999, Marcel Dekker, New York, the contents of each of which are incorporated herein by reference. Except for any conventional carriers that are incompatible with the disclosed compounds of the present invention, such as those that may produce any undesirable biological effects or that may interact in a deleterious manner with any other ingredient in the pharmaceutically acceptable composition, it is contemplated that their use is within the scope of the present invention.

[0097] The pharmaceutical compositions disclosed herein are prepared using techniques and methods known to those skilled in the art. A description of some common methods in the art can be found in Remington's Pharmaceutical Sciences (Mack Publishing Company).

[0098] Thus, in another aspect, the present invention relates to a process for preparing a pharmaceutical composition comprising a compound disclosed herein and a pharmaceutically acceptable excipient, vehicle, carrier, or combination thereof, the process comprising mixing the ingredients. Pharmaceutical compositions comprising a compound disclosed herein can be prepared by mixing at, for example, ambient temperature and atmospheric pressure.

[0099] The compounds disclosed herein are generally formulated into dosage forms suitable for administration to a patient via a desired route. For example, dosage forms include those suitable for the following routes of administration: (1) oral administration, such as tablets, capsules, caplets, pills, lozenges, powders, syrups, elixirs, suspensions, solutions, emulsions, sachets, and cachets; (2) parenteral administration, such as sterile solutions, suspensions, and reconstituted powders; (3) transdermal administration, such as transdermal patches; (4) rectal administration, such as suppositories; (5) inhalation, such as aerosols, solutions, and dry powders; and (6) topical administration, such as creams, ointments, lotions, solutions, pastes, sprays, foams, and gels.

[0100] In one embodiment, the compounds disclosed herein can be formulated as oral dosage forms. In another embodiment, the compounds disclosed herein can be formulated as inhalation dosage forms. In another embodiment, the compounds disclosed herein can be formulated as nasal dosage forms. In yet another embodiment, the compounds disclosed herein can be formulated as transdermal dosage forms. In yet another embodiment, the compounds disclosed herein can be formulated as topical dosage forms.

[0101] The pharmaceutical compositions provided herein can be provided as compressed tablets, tablets, chewable lozenges, fast-dissolving tablets, composite compressed tablets, or enteric-coated, sugar-coated, or film-coated tablets. Enteric-coated tablets are compressed tablets coated with a substance that resists the effects of gastric acid but dissolves or disintegrates in the intestines, thereby protecting the active ingredients from the acidic environment of the stomach. Enteric coatings include, but are not limited to, fatty acids, fats, phenyl salicylate, waxes, shellac, ammoniated shellac, and cellulose acetate phthalate. Sugar-coated tablets are compressed tablets surrounded by a sugar coating, which helps mask unpleasant tastes or odors and prevents tablet oxidation. Film-coated tablets are compressed tablets covered with a thin layer or film of a water-soluble substance. Film coatings include, but are not limited to, hydroxyethylcellulose, sodium carboxymethylcellulose, polyethylene glycol 4000, and cellulose acetate phthalate. Film coatings impart the same general properties as sugar coatings. Composite compressed tablets are compressed tablets produced through more than one compression cycle, including multilayer tablets and press-coated or dry-coated tablets.

[0102] Tablet dosage forms can be prepared from the active ingredient in powder, crystal or granular form alone or in combination with one or more carriers or excipients described herein, including binders, disintegrants, controlled release polymers, lubricants, diluents and / or colorants. Flavoring agents and sweeteners are particularly useful in forming chewable tablets and lozenges.

[0103] The pharmaceutical composition provided by the present invention can be provided in soft capsules or hard capsules, which can be prepared from gelatin, methylcellulose, starch or calcium alginate. The hard gelatin capsule, also known as dry-filled capsule (DFC), consists of two sections, one section inserted into the other, thus completely encapsulating the active ingredient. Soft elastic capsules (SEC) are soft, spherical shells, such as gelatin shells, which are plasticized by adding glycerol, sorbitol or similar polyols. The soft gelatin shells can contain preservatives to prevent microbial growth. Suitable preservatives are those as described in the present invention, including methylparaben and propylparaben, and sorbic acid. The liquid, semisolid and solid dosage forms provided by the present invention can be encapsulated in capsules. Suitable liquid and semisolid dosage forms include solutions and suspensions in propylene carbonate, vegetable oils or triglycerides. Capsules containing such solutions can be prepared as described in U.S. Pat. Nos. 4,328,245; 4,409,239 and 4,410,545. The capsules may also be coated as known to those skilled in the art to improve or sustain dissolution of the active ingredient.

[0104] The pharmaceutical compositions provided herein can be provided in liquid and semisolid dosage forms, including emulsions, solutions, suspensions, elixirs, and syrups. Emulsions are two-phase systems in which one liquid is completely dispersed in another liquid in the form of small globules, which can be oil-in-water or water-in-oil. Emulsions can include pharmaceutically acceptable non-aqueous liquids and solvents, emulsifiers, and preservatives. Suspensions can include pharmaceutically acceptable suspending agents and preservatives. Aqueous alcoholic solutions can include pharmaceutically acceptable acetals, such as di(lower alkyl) acetals of lower alkyl aldehydes, such as acetaldehyde diethyl acetal; and water-soluble solvents having one or more hydroxyl groups, such as propylene glycol and ethanol. Elixirs are clear, sweet-tasting hydroalcoholic solutions. Syrups are concentrated aqueous solutions of sugars, such as sucrose, and can also contain preservatives. For liquid dosage forms, for example, solutions in polyethylene glycol can be diluted with a sufficient amount of a pharmaceutically acceptable liquid carrier, such as water, for accurate and convenient administration.

[0105] Other useful liquid and semisolid dosage forms include, but are not limited to, those comprising the active ingredient provided herein and a secondary mono- or poly-alkylene glycol, including 1,2-dimethoxymethane, diethylene glycol dimethyl ether, triethylene glycol dimethyl ether, tetraethylene glycol dimethyl ether, polyethylene glycol-350-dimethyl ether, polyethylene glycol-550-dimethyl ether, polyethylene glycol-750-dimethyl ether, wherein 350, 550, and 750 refer to the approximate average molecular weight of the polyethylene glycol. These formulations may further include one or more antioxidants, such as butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), propyl gallate, vitamin E, hydroquinone, hydroxycoumarin, ethanolamine, lecithin, cephalin, ascorbic acid, malic acid, sorbitol, phosphoric acid, bisulfite, sodium metabisulfite, thiodipropionic acid and its esters and dithiocarbamates.

[0106] Where appropriate, dosage unit formulations for oral administration can be microencapsulated.Delayed or sustained-release compositions can also be prepared, for example, by coating or embedding particulate material in polymers, wax, or the like.

[0107] The oral pharmaceutical composition provided by the present invention can also be provided in the form of liposomes, micelles, microspheres or nanosystems. Micellar dosage forms can be prepared using the method described in US Pat. No. 6,350,458.

[0108] The pharmaceutical compositions provided herein can be provided in non-effervescent or effervescent granules and powders for reconstitution into liquid dosage forms. Pharmaceutically acceptable carriers and excipients used in non-effervescent granules or powders can include diluents, sweeteners, and wetting agents. Pharmaceutically acceptable carriers and excipients used in effervescent granules or powders can include organic acids and carbon dioxide sources.

[0109] Coloring and flavoring agents can be used in all of the above dosage forms.

[0110] The compounds disclosed herein can also be combined with soluble polymers as targeted drug carriers. Such polymers include polyvinylpyrrolidone, pyran copolymer, polyhydroxypropylmethacrylamide-phenol, polyhydroxyethylaspartamidephenol, or polyethylene glycol polylysine substituted with palmitoyl residues. In addition, the compounds disclosed herein can be combined with a class of biodegradable polymers used in achieving controlled release of drugs, such as crosslinked or amphiphilic block copolymers of polylactic acid, poly-ε-caprolactone, polyhydroxybutyric acid, polyorthoesters, polyacetals, polydihydropyrans, polycyanoacrylates, and hydrogels.

[0111] The pharmaceutical compositions provided herein can be formulated as immediate or modified release dosage forms, including delayed-, sustained-, pulsed-, controlled-, targeted-, and programmed-release forms.

[0112] The pharmaceutical composition provided by the present invention can be co-formulated with other active ingredients that do not impair the intended therapeutic effect, or co-formulated with substances that supplement the intended effect.

[0113] Pharmaceutical composition provided by the invention can be administered parenterally by injection, infusion or implantation, for local or systemic administration. Parenteral administration as used in the present invention includes intravenous, intraarterial, intraperitoneal, intrathecal, intraventricular, intraurethral, ​​intrasternal, intracranial, intramuscular, intrasynovial and subcutaneous administration.

[0114] Pharmaceutical composition provided by the invention can be mixed with any dosage form suitable for parenteral administration, including solution, suspension, emulsion, micelle, liposome, microsphere, nanometer system and the solid form that is suitable for making solution or suspension in liquid before injection.Such dosage form can be prepared (referring to Remington:The Science and Practice of Pharmacy, the same) according to conventional method known to those skilled in the art of pharmaceutical science.

[0115] Pharmaceutical compositions intended for parenteral administration may include one or more pharmaceutically acceptable carriers and excipients, including, but not limited to, aqueous carriers, water-miscible carriers, non-aqueous carriers, antimicrobial agents or preservatives against microbial growth, stabilizers, solubility enhancers, isotonicity agents, buffers, antioxidants, local anesthetics, suspending and dispersing agents, wetting or emulsifying agents, complexing agents, sequestering or chelating agents, antifreezes, cryoprotectants, thickeners, pH adjusters, and inert gases.

[0116] Suitable aqueous vehicles include, but are not limited to, water, saline, physiological saline or phosphate buffered saline (PBS), sodium chloride injection, Ringers injection, isotonic dextrose injection, sterile water injection, dextrose, and lactated Ringers injection. Non-aqueous vehicles include, but are not limited to, fixed oils of plant origin, castor oil, corn oil, cottonseed oil, olive oil, peanut oil, peppermint oil, safflower oil, sesame oil, soybean oil, hydrogenated vegetable oils, medium-chain triglycerides of hydrogenated soybean oil and coconut oil, and palm seed oil. Water-miscible vehicles include, but are not limited to, ethanol, 1,3-butylene glycol, liquid polyethylene glycols (e.g., polyethylene glycol 300 and polyethylene glycol 400), propylene glycol, glycerol, N-methyl-2-pyrrolidone, N,N-dimethylacetamide, and dimethyl sulfoxide.

[0117] Suitable antimicrobial agents or preservatives include, but are not limited to, phenol, cresol, mercurials, benzyl alcohol, chlorobutanol, methyl and propyl parabens, thimerosal, benzalkonium chloride (e.g., benzethonium chloride), methyl and propyl parabens, and sorbic acid. Suitable isotonic agents include, but are not limited to, sodium chloride, glycerol, and dextrose. Suitable buffers include, but are not limited to, phosphates and citrates. Suitable antioxidants are those described herein, including bisulfites and sodium metabisulfite. Suitable local anesthetics include, but are not limited to, procaine hydrochloride. Suitable suspending and dispersing agents are those described herein, including sodium carboxymethylcellulose, hydroxypropyl methylcellulose, and polyvinylpyrrolidone. Suitable emulsifiers include those described herein, including polyoxyethylene sorbitan monolaurate, polyoxyethylene sorbitan monooleate 80, and triethanolamine oleate. Suitable sequestrants or chelating agents include, but are not limited to EDTA. Suitable pH adjusting agents include, but are not limited to, sodium hydroxide, hydrochloric acid, citric acid, and lactic acid. Suitable complexing agents include, but are not limited to, cyclodextrins, including α-cyclodextrin, β-cyclodextrin, hydroxypropyl-β-cyclodextrin, sulfobutyl ether-β-cyclodextrin, and sulfobutyl ether 7-β-cyclodextrin ( CyDex, Lenexa, KS).

[0118] The pharmaceutical compositions provided herein can be formulated for single or multiple dose administration. The single dose formulations are packaged in ampoules, vials, or syringes. The multiple dose parenteral formulations must contain an antimicrobial agent at a bacteriostatic or fungistatic concentration. All parenteral formulations must be sterile, as known and practiced in the art.

[0119] In one embodiment, the pharmaceutical composition is provided as a ready-to-use sterile solution. In another embodiment, the pharmaceutical composition is provided as a sterile dry soluble product, including lyophilized powders and hypodermic tablets, which are reconstituted with a vehicle prior to use. In yet another embodiment, the pharmaceutical composition is formulated as a ready-to-use sterile suspension. In yet another embodiment, the pharmaceutical composition is formulated as a sterile dry insoluble product that is reconstituted with a vehicle prior to use. In yet another embodiment, the pharmaceutical composition is formulated as a ready-to-use sterile emulsion.

[0120] The pharmaceutical compositions disclosed herein can be formulated as immediate or modified release dosage forms, including delayed-, sustained-, pulsed-, controlled-, targeted-, and programmed-release forms.

[0121] The pharmaceutical composition can be formulated as a suspension, solid, semisolid, or thixotropic liquid for administration as an implanted depot. In one embodiment, the pharmaceutical composition disclosed herein is dispersed in a solid inner matrix, which is surrounded by an outer polymeric membrane that is insoluble in body fluids but allows the active ingredient in the pharmaceutical composition to diffuse through.

[0122] Suitable inner matrices include polymethyl methacrylate, polybutyl methyl acrylate, plasticized or unplasticized polyvinyl chloride, plasticized nylon, plasticized polyethylene terephthalate, plasticized polyethylene terephthalate, natural rubber, polyisoprene, polyisobutylene, polybutadiene, polyethylene, ethylene-vinyl acetate copolymers, silicone rubber, polydimethylsiloxane, silicone carbonate copolymers, hydrophilic polymers such as hydrogels of esters of acrylic acid and methacrylic acid, collagen, cross-linked polyvinyl alcohol, and partially hydrolyzed polyvinyl acetate of coach.

[0123] Suitable outer polymeric films include polyethylene, polypropylene, ethylene / propylene copolymers, ethylene / ethyl acrylate copolymers, ethylene / vinyl acetate copolymers, silicone rubber, polydimethylsiloxane, neoprene, chlorinated polyethylene, polyvinyl chloride, copolymers of chlorinated ethylene and vinyl acetate, vinylidene chloride, ethylene and propylene, ionomer polyethylene terephthalate, butyl rubber epichlorohydrin rubber, ethylene / vinyl alcohol copolymers, ethylene / vinyl acetate / vinyl alcohol terpolymers, and ethylene / vinyloxyethanol copolymers.

[0124] On the other hand, the pharmaceutical compositions disclosed herein can be formulated into any dosage form suitable for inhalation administration to a patient, such as a dry powder, an aerosol, a suspension or a solution composition. In one embodiment, the pharmaceutical compositions disclosed herein can be formulated into a dosage form suitable for inhalation administration to a patient using a dry powder. In another embodiment, the pharmaceutical compositions disclosed herein can be formulated into a dosage form suitable for inhalation administration to a patient via a nebulizer. Dry powder compositions delivered to the lungs by inhalation typically comprise a finely powdered compound disclosed herein and one or more finely powdered pharmaceutically acceptable excipients. Pharmaceutically acceptable excipients particularly suitable for use as dry powders are known to those skilled in the art and include lactose, starch, mannitol, and mono-, di- and polysaccharides. Fine powders can be prepared, for example, by micronization and grinding. Generally, size-reduced (e.g., micronized) compounds can be prepared by a D 50 values ​​(e.g., measured by laser diffraction).

[0125] Aerosols can be prepared by suspending or dissolving the compounds disclosed herein in a liquefied propellant. Suitable propellants include chlorinated hydrocarbons, hydrocarbons, and other liquefied gases. Representative propellants include: trichlorofluoromethane (propellant 11), dichlorofluoromethane (propellant 12), dichlorotetrafluoroethane (propellant 114), tetrafluoroethane (HFA-134a), 1,1-difluoroethane (HFA-152a), difluoromethane (HFA-32), pentafluoroethane (HFA-12), heptafluoropropane (HFA-227a), perfluoropropane, perfluorobutane, perfluoropentane, butane, isobutane, and pentane. Aerosols containing the compounds disclosed herein are typically administered to patients via a metered dose inhaler (MDI). Such devices are known to those skilled in the art.

[0126] Aerosols may contain additional pharmaceutically acceptable excipients that can be used with MDIs, such as surfactants, lubricants, co-solvents, and other excipients to improve the physical stability of the formulation, improve valve characteristics, improve solubility, or improve taste.

[0127] Pharmaceutical compositions suitable for transdermal administration can be prepared as discontinuous patches intended to remain in close contact with the patient's epidermis for an extended period of time. For example, the active ingredient can be delivered from the patch by iontophoresis as generally described in Pharmaceutical Research, 3(6), 318 (1986).

[0128] The pharmaceutical composition that is suitable for topical administration can be configured to ointment, cream, suspension, lotion, powder, solution, paste, gel, spray, aerosol or oil.For example, ointment, cream and gel can configure with water or oil base, and applicable thickener and / or gel and / or solvent.Such matrix can comprise, water, and / or oil such as liquid liquid paraffin and vegetable oil (such as peanut oil or castor oil), or solvent such as polyethylene glycol.The thickener and gel that use according to matrix properties comprise soft paraffin, aluminum stearate, cetearyl alcohol, polyethylene glycol, lanolin, beeswax, carboxyvinyl polyol and cellulose derivative, and / or glyceryl monostearate and / or nonionic emulsifier.

[0129] Lotions may be formulated with an aqueous or oily base, and generally also contain one or more emulsifying agents, stabilizing agents, dispersing agents, suspending agents or thickening agents.

[0130] Powders for external use may be formulated in the presence of any suitable powder base such as talc, lactose or starch. Drops may be formulated with an aqueous or non-aqueous base containing one or more dispersants, solubilizers, suspending agents or preservatives.

[0131] Topical formulations can be administered by application to the affected area once or more daily; occlusive dressings covering the skin are preferred. Adhesive reservoir systems allow for continuous or prolonged administration.

[0132] For treatment of the eye, or other organs such as the mouth and skin, the compositions can be applied as a topical ointment or cream. When formulated as an ointment, the compounds disclosed herein can be used with either a paraffinic or a water-soluble ointment base. Alternatively, the compounds disclosed herein can be formulated into a cream with an oil-in-water cream base or an oil-in-water base.

[0133] In one embodiment, the treatment method disclosed herein comprises administering a safe and effective amount of a compound of the present invention or a pharmaceutical composition comprising the compound of the present invention to a patient in need thereof. Various embodiments disclosed herein include methods for treating the above-mentioned diseases by administering a safe and effective amount of a compound of the present invention or a pharmaceutical composition comprising the compound of the present invention to a patient in need thereof.

[0134] In one embodiment, the compounds disclosed herein or pharmaceutical compositions comprising the compounds disclosed herein can be administered by any suitable route of administration, including systemic administration and topical administration. Systemic administration includes oral administration, parenteral administration, transdermal administration, and rectal administration. Typical parenteral administration refers to administration by injection or infusion, including intravenous, intramuscular, and subcutaneous injection or infusion. Topical administration includes application to the skin, as well as intraocular, ear, vaginal, inhalation, and intranasal administration. In one embodiment, the compounds disclosed herein or pharmaceutical compositions comprising the compounds disclosed herein can be administered orally. In another embodiment, the compounds disclosed herein or pharmaceutical compositions comprising the compounds disclosed herein can be administered by inhalation. In another embodiment, the compounds disclosed herein or pharmaceutical compositions comprising the compounds disclosed herein can be administered intranasally.

[0135] In one embodiment, the compounds disclosed herein or pharmaceutical compositions comprising the compounds disclosed herein can be administered once or several times at different time intervals over a specified period of time according to a dosing regimen. For example, the drug may be administered once, twice, three times, or four times a day. In one embodiment, the drug is administered once a day. In another embodiment, the drug is administered twice a day. The drug may be administered until the desired therapeutic effect is achieved or the desired therapeutic effect is maintained indefinitely. The appropriate dosing regimen for the compounds disclosed herein or pharmaceutical compositions comprising the compounds disclosed herein depends on the pharmacokinetic properties of the compound, such as dilution, distribution, and half-life, which can be determined by a skilled person. In addition, the appropriate dosing regimen for the compounds disclosed herein or pharmaceutical compositions comprising the compounds disclosed herein, including the duration of implementation of the regimen, depends on the disease being treated, the severity of the disease being treated, the age and physical condition of the patient being treated, the medical history of the patient being treated, the nature of the concurrent therapy, the desired therapeutic effect, and other factors within the knowledge and experience of the skilled person. Such skilled persons should also understand that the dosing regimen may require adjustment to the individual patient's response to the dosing regimen or as the individual patient's needs change over time.

[0136] The compounds disclosed herein can be administered simultaneously with, before, or after one or more other therapeutic agents. The compounds disclosed herein can be administered separately with other therapeutic agents via the same or different routes of administration, or in the form of a pharmaceutical composition.

[0137] For individuals weighing about 50-70 kg, the pharmaceutical compositions and combinations disclosed herein can be in unit dosage form containing about 1-1000 mg, or about 1-500 mg, or about 1-250 mg, or about 1-150 mg, or about 0.5-100 mg, or about 1-50 mg of active ingredient. The therapeutically effective amount of the compound, pharmaceutical composition, or combination thereof depends on the species, weight, age, and individual condition of the individual, the disorder or disease being treated, or its severity. A physician, clinician, or veterinarian with ordinary skill can readily determine the effective amount of each active ingredient required to prevent, treat, or inhibit the progression of a disorder or disease.

[0138] The dosage characteristics cited above have been demonstrated in vitro and in vivo using advantageous mammals (e.g., mice, rats, dogs, monkeys) or isolated organs, tissues, and specimens thereof. The compounds disclosed herein are administered in vitro in the form of solutions, e.g., aqueous solutions, and in vivo in the form of suspensions or aqueous solutions, either enterally, parenterally, or particularly intravenously.

[0139] In one embodiment, a therapeutically effective dose of a compound disclosed herein is from about 0.1 mg to about 2,000 mg per day. A pharmaceutical composition thereof should provide a dose of from about 0.1 mg to about 2,000 mg of the compound. In a specific embodiment, a pharmaceutical dosage unit form is prepared that provides from about 1 mg to about 2,000 mg, from about 10 mg to about 1,000 mg, from about 20 mg to about 500 mg, or from about 25 mg to about 250 mg of the active ingredient, or a combination of active ingredients, per dosage unit form. In a specific embodiment, a pharmaceutical dosage unit form is prepared that provides from about 10 mg, 20 mg, 25 mg, 50 mg, 100 mg, 250 mg, 500 mg, 1000 mg, or 2000 mg of the active ingredient.

[0140] In addition, the compounds disclosed in the present invention can be administered in the form of prodrugs. In the present invention, the "prodrug" of the compounds disclosed in the present invention is a functional derivative that can ultimately release the compounds disclosed in the present invention in vivo when administered to a patient. When administering the compounds disclosed in the present invention in the form of prodrugs, those skilled in the art may implement one or more of the following methods: (a) changing the onset time of the compound in vivo; (b) changing the duration of the compound's action in vivo; (c) changing the compound's transport or distribution in vivo; (d) changing the compound's solubility in vivo; and (e) overcoming the side effects or other difficulties faced by the compound. Typical functional derivatives used to prepare prodrugs include variants of the compound that are chemically or enzymatically cleaved in vivo. These variants, including those for preparing phosphates, amides, esters, thioesters, carbonates, and carbamates, are well known to those skilled in the art.

[0141] Definitions and Explanations of Terms

[0142] Unless otherwise defined, all technical and scientific terms herein have the same meanings as commonly understood by persons skilled in the art to which the claimed subject matter belongs. Unless otherwise indicated, all patents, patent applications, and publications cited herein are incorporated by reference in their entirety. If multiple definitions of a term are used herein, the definitions in this section shall prevail.

[0143] It should be understood that the above brief description and the following detailed description are exemplary and for explanation only, and do not limit the subject matter of this application. In this application, unless otherwise indicated, the use of "or" or "alternatively" means "and / or". In addition, the use of the term "including" and other forms such as "comprising", "including" and "containing" are not limiting.

[0144] The term "alkyl" refers to a straight or branched chain alkyl group having 1 to 6 carbon atoms, for example, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, sec-butyl, pentyl, neopentyl.

[0145] The term "alkoxy" refers to a straight or branched alkoxy group having 1 to 6 carbon atoms, for example, methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, tert-butoxy, sec-butoxy.

[0146] The term "halogen" refers to fluorine, chlorine, bromine, or iodine, preferably fluorine, chlorine, or bromine.

[0147] The term "aryl" is understood to mean preferably a monovalent aromatic or partially aromatic monocyclic, bicyclic or tricyclic hydrocarbon ring having 6 to 20 carbon atoms, preferably "C 6-14 Aryl". The term "C 6-1 "Aryl" is understood to mean preferably a monovalent aromatic or partially aromatic monocyclic, bicyclic or tricyclic hydrocarbon ring ("C 6-14 or a ring having 9 carbon atoms ("C9 aryl"), for example indanyl or indenyl, or a ring having 10 carbon atoms ("C 10 aryl) such as tetrahydronaphthyl, dihydronaphthyl or naphthyl, or a ring having 13 carbon atoms ("C 13 aryl), such as fluorenyl, or a ring having 14 carbon atoms ("C 14 "aryl"), for example anthracenyl.

[0148] The term "heteroaryl" should be understood as containing 5-20 ring atoms, 5-14 ring atoms, or 5-12 ring atoms, or 5-10 ring atoms, or 5-6 ring atoms. Monocyclic, bicyclic and tricyclic ring systems, wherein at least one ring system is aromatic, and at least one ring system contains one or more heteroatoms (e.g., N, O, S, Se, etc.), wherein each ring system contains a ring consisting of 5-7 atoms and has one or more connection points connected to the rest of the molecule. The heteroaryl group is optionally substituted with one or more substituents described herein. In some embodiments, the heteroaryl group consisting of 5-10 atoms contains 1, 2, 3 or 4 heteroatoms independently selected from O, S, Se and N. In other embodiments, the heteroaryl group consisting of 5-6 atoms contains 1, 2, 3 or 4 heteroatoms independently selected from O, S, Se and N.

[0149] Examples of monocyclic heteroaryl groups include, but are not limited to, thienyl, furyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, thiadiazolyl, thia-4H-pyrazolyl, and the like, and benzo derivatives thereof, such as benzofuranyl, benzothienyl, benzoxazolyl, benzisoxazolyl, benzimidazolyl, benzotriazolyl, indazolyl, indolyl, isoindolyl, etc.; or pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, etc., and benzo derivatives thereof, such as quinolyl, quinazolinyl, isoquinolyl, etc.; or acinyl, indolizinyl, purinyl, etc., and benzo derivatives thereof; or cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, naphthyridinyl, pteridinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, etc.

[0150] The term "heterocyclyl" refers to a monocyclic, bicyclic or tricyclic ring system in which one or more ring atoms are independently optionally substituted with heteroatoms, the ring may be fully saturated or contain one or more degrees of unsaturation, but is not aromatic, and has one or more points of attachment to other molecules. One or more ring hydrogen atoms may be independently unsubstituted or substituted with one or more substituents described herein. In some embodiments, the "heterocyclyl" is a monocyclic ring of 3-7 atoms or a bicyclic ring of 7-10 atoms, containing 1-5, preferably 1-3, heteroatoms selected from N, O, S and Se. In particular, the heterocyclic group may include, but is not limited to, a 4-membered ring such as azetidinyl, oxetanyl; a 5-membered ring such as tetrahydrofuranyl, dioxolyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, pyrrolinyl; or a 6-membered ring such as tetrahydropyranyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl or trithianyl; or a 7-membered ring such as diazepanyl. Optionally, the heterocyclic group may be benzo-fused. The heterocyclic group may be bicyclic, for example, but not limited to, a 5,5-membered ring such as hexahydrocyclopenta [c] pyrrole-2 (1H) -yl ring, or a 5,6-membered bicyclic ring such as hexahydropyrrolo [1,2-a] pyrazine-2 (1H) -yl ring. The ring containing the nitrogen atom may be partially unsaturated, i.e., it may contain one or more double bonds, such as, but not limited to, 2,5-dihydro-1H-pyrrolyl, 4H-[1,3,4]thiadiazinyl, 4,5-dihydrooxazolyl or 4H-[1,4]thiazinyl, or it may be benzo-fused, such as, but not limited to, dihydroisoquinolinyl, 1,3-benzoxazolyl, 1,3-benzodioxolyl.

[0151] Unless otherwise indicated, heterocyclyl and heteroaryl include all possible isomeric forms thereof, such as positional isomers thereof. Thus, for some illustrative, non-limiting examples, pyridyl or pyridinylene include pyridin-2-yl, pyridin-2-ylene, pyridin-3-yl, pyridin-3-ylene, pyridin-4-ylene, and pyridin-4-ylene; thienyl or thienylene include thien-2-yl, thien-2-ylene, thien-3-ylene, and thien-3-ylene.

[0152] As used herein, the term "treat" and other similar synonyms include alleviating, reducing or ameliorating the symptoms of a disease or condition, preventing other symptoms, ameliorating or preventing the underlying metabolic causes of symptoms, inhibiting the disease or condition, such as preventing the development of the disease or condition, alleviating the disease or condition, making the disease or condition better, alleviating the symptoms caused by the disease or condition, or stopping the symptoms of the disease or condition. In addition, the term includes the purpose of prevention. The term also includes obtaining a therapeutic effect and / or a prophylactic effect. The therapeutic effect refers to curing or improving the underlying disease being treated. In addition, the cure or improvement of one or more physiological symptoms associated with the underlying disease is also a therapeutic effect, for example, although the patient may still be affected by the underlying disease, the patient's condition is observed to improve. In terms of prophylactic effect, the composition can be administered to a patient at risk for a particular disease, or even if a diagnosis of the disease has not yet been made, the composition can be administered to a patient who has one or more physiological symptoms of the disease.

[0153] As used herein, the terms "effective amount," "therapeutically effective amount," or "pharmaceutically effective amount" refer to an amount of at least one agent or compound sufficient to provide some relief to some degree from one or more symptoms of the disease or condition being treated. This can result in a reduction and / or alleviation of signs, symptoms, or causes of disease, or any other desired change in a biological system. For example, a therapeutically effective amount is the amount of a composition comprising a compound disclosed herein that provides a clinically significant alleviation of symptoms. Techniques such as dose escalation studies can be used to determine the effective amount appropriate for any individual case.

[0154] As used herein, the terms "administer," "administer," "dosing," and the like refer to methods that enable a compound or composition to be delivered to the desired site of biological action. These methods include, but are not limited to, oral routes, intraduodenal routes, parenteral injection (including intravenous, subcutaneous, intraperitoneal, intramuscular, intraarterial injection or infusion), topical, and rectal administration. Those skilled in the art are familiar with administration techniques that can be used for the compounds and methods described herein. In preferred embodiments, the compounds and compositions discussed herein are administered orally.

[0155] The term "acceptable" with respect to a formulation, composition or ingredient, as used herein, means having no long-term detrimental effect on the general health of the subject being treated.

[0156] As used herein, the term "pharmaceutically acceptable" refers to a substance (such as a carrier or diluent) that does not affect the biological activity or properties of the compounds of the present invention and is relatively non-toxic, that is, the substance can be administered to a subject without causing adverse biological reactions or interacting in an adverse manner with any components contained in the composition.

[0157] As used herein, the term "pharmaceutical composition" refers to a biologically active compound optionally mixed with at least one pharmaceutically acceptable chemical component, including but not limited to carriers, stabilizers, diluents, dispersants, suspending agents, thickeners and / or excipients.

[0158] As used herein, the term "carrier" refers to relatively nontoxic chemical compounds or agents that facilitate the introduction of a compound into cells or tissues.

[0159] As used herein, the term "pharmaceutically acceptable salt" refers to a salt that retains the biological effectiveness of the free acid and free base of the specified compound and has no adverse biological or other effects. Examples of pharmaceutically acceptable salts include, but are not limited to, inorganic or organic acid salts of basic residues (such as amines), alkaline or organic salts of acidic residues (such as carboxylic acids), and the like. Pharmaceutically acceptable salts include conventional non-toxic salts or quaternary ammonium salts of the parent compound formed from, for example, non-toxic inorganic or organic acids. For example, these conventional non-toxic salts include those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, phosphoric acid, nitric acid, and the like; and salts prepared from organic acids such as acetic acid, propionic acid, succinic acid, glycolic acid, lactic acid, malic acid, tartaric acid, citric acid, fumaric acid, methanesulfonic acid, toluenesulfonic acid, salicylic acid, p-aminobenzenesulfonic acid, and the like.

[0160] Pharmaceutically acceptable salts of the present invention can be synthesized from the parent compound containing alkaline or acidic moieties by conventional chemical methods. Usually, this salt can be prepared by reacting the free acid or alkaline form of these compounds with stoichiometric appropriate alkali or acid in water or an organic solvent or a mixture of the two.

[0161] In the description of this specification, the reference terms "one embodiment", "some embodiments", "example", "specific example", or "some examples" mean that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described can be combined in any one or more embodiments or examples in a suitable manner. In addition, those skilled in the art can combine and combine different embodiments or examples described in this specification and features of different embodiments or examples without contradiction.

[0162] Compound preparation route:

[0163] The first step of the reaction:

[0164] To a 50 mL round-bottom flask, aryl bromide A (1 mmol, 1.0 equiv), aryl boronic acid compound B (1.5 mmol, 1.5 equiv), tetrakistriphenylphosphine palladium (0.05 mmol, 0.05 equiv), potassium carbonate (3 mmol, 3 equiv), and 10 mL of 1,4-dioxane aqueous solution (1,4-dioxane:H2O = 10:1) were added sequentially. Under nitrogen, the mixture was stirred at 100°C for 3 hours. After completion, 20 mL of water was added to terminate the reaction. The reaction solution was extracted with 100 mL of ethyl acetate three times. The organic phases were combined and washed once with 30 mL of saturated brine. The organic phases were dried over anhydrous sodium sulfate and concentrated under reduced pressure to remove the solvent, yielding the crude product. Purification by flash column chromatography afforded ester C.

[0165] Second step of reaction:

[0166] Compound C was dissolved in a mixture of tetrahydrofuran and methanol (volume ratio 1:1), and then an equal volume of aqueous sodium hydroxide solution was added dropwise to a final concentration of 1M. The mixture was stirred at room temperature for 5 hours for hydrolysis to obtain compound D. The tetrahydrofuran and methanol were removed by rotary evaporation, and the pH was adjusted to 5 with 1M hydrochloric acid to precipitate compound D. Product D was extracted with 120mL of ethyl acetate, and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to remove the solvent to obtain a crude product. Carboxylic acid D was purified by flash column chromatography.

[0167] Reaction step 3:

[0168] In a 50 mL reaction flask, carboxylic acid D (1.0 equivalent) was dissolved in an appropriate amount of solvent, and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (2.0 equivalents) and N,N-diisopropylethylamine (4.0 equivalents) were added. After stirring at room temperature for 15 minutes, compound E (1.1 equivalents) was added and stirring was continued at room temperature overnight. After completion of the reaction, the reaction solution was poured into 200 mL of ethyl acetate, and the organic phase was washed twice with deionized water and saturated brine. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to remove the solvent to obtain the crude product. Purification by flash column chromatography gave compound F.

[0169] Reaction step 4:

[0170] Compound F was dissolved in tetrahydrofuran, and an equal volume of aqueous lithium hydroxide solution was added to give a final concentration of 1 M lithium hydroxide. The hydrolysis reaction was stirred at room temperature, and the reaction progress was monitored by thin-layer chromatography. After the reaction was complete, the tetrahydrofuran was removed by rotary evaporation, and the pH was adjusted to 5 with 1 M hydrochloric acid to precipitate compound G. Product G was extracted with 120 mL of ethyl acetate, and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to remove the solvent to obtain crude product G.

[0171] Reaction step 5:

[0172] Crude Compound G was dissolved in 1,4-dioxane, and a 1,4-dioxane solution of hydrochloric acid was slowly added dropwise at 0°C to a final concentration of 2 M. The reaction solution was then slowly warmed to room temperature, and the reaction progress was monitored by HPLC-MS. After the reaction was complete, the 1,4-dioxane was removed by rotary evaporation, and the crude product was dissolved in a trace amount of N,N-dimethylformamide and purified by reverse-phase preparative chromatography to obtain product 20.

[0173] The preparation of the following compounds refers to the compound preparation route. 1 The H NMR characterization results are as follows:

[0174] Compound 1

[0175] 1 H NMR (400MHz, DMSO-d6) δ10.32(s,1H),9.15(s,1H),8.87(d,J=6.5Hz,1H),7.98( d,J=8.3Hz,2H),7.77(dd,J=20.6,7.9Hz,4H),7.50(t,J=7.6Hz,2H),7.42(t,J=7 .3Hz,1H),4.63(dd,J=12.6,5.9Hz,1H),4.42(s,1H),3.58(dd,J=6.8,3.4Hz,1H ),3.48(dd,J=11.5,4.3Hz,1H),2.69(dt,J=13.2,8.2Hz,1H),2.34–2.05(m,1H).

[0176] Compound 2

[0177] 11H NMR (400 MHz, DMSO-d6) δ 8.56 (d, J = 6.1 Hz, 1H), 7.50–7.24 (m, 4H), 7.12 (t, J = 7.1 Hz, 1H), 6.94 (d, J = 7.8 Hz, 2H), 6.82 (d, J = 8.6 Hz, 1H), 4.29 (t, J = 8.9 Hz, 1H), 3.59 (s, 1H), 3.51 (s, 2H), 3.41–3.25 (m, 1H), 3.09–2.85 (m, 1H), 2.60–2.43 (m, 1H), 1.86 (dd, J = 21.2, 9.2 Hz, 1H).

[0178] Compound 3

[0179] 1 1H NMR (400 MHz, DMSO-d6) δ 9.18 (d, J = 6.6 Hz, 1H), 8.57 (s, 1H), 8.20 (s, 1H), 8.03 (dd, J = 22.7, 8.6 Hz, 3H), 7.88 (d, J = 8.6 Hz, 1H), 7.64 (d, J = 8.5 Hz, 1H), 7.57 (d, J = 1.6 Hz, 1H), 7.11 (d, J = 8.6 Hz, 1H), 4.68 (dd, J = 13.5, 6.8 Hz, 1H), 4.37 (t, J = 8.9 Hz, 1H), 3.86 (s, 3H), 3.45 - 3.42 (m, 2H), 2.71 (dt, J = 13.1, 7.9 Hz, 1H), 2.43 - 2.16 (m, 1H), 2.06 (s, 9H), 1.75 (s, 6H).

[0180] Compound 4

[0181] 1 1H NMR (400 MHz, DMSO-d6) δ 8.94 (d, J = 6.6 Hz, 1H), 8.49 (s, 1H), 8.24 (s, 1H), 8.12–8.03 (m, 2H), 8.00–7.91 (m, 2H), 7.​​​​​

[0183] 1 1H NMR (400 MHz, MeOD) δ 8.44 (s, 1H), 8.16 (s, 1H), 8.11–8.02 (m, 2H), 7.91 (dd, J = 18.3, 8.4 Hz, 2H), 7.79 (s, 1H), 7.58 (d, J = 7.3 Hz, 1H), 7.46 (dt, J = 15.1, 7.7 Hz, 2H), 4.80–4.68 (m, 1H), 4.57 (t, J = 8.6 Hz, 1H), 3.76 (dd, J = 12.1, 7.2 Hz, 1H), 3.59 (dd, J = 12.0, 5.3 Hz, 1H), 3.37 (s, 1H), 3.09–2.81 (m, 1H), 2.67–2.23 (m, 1H), 1.43 (s, 9H).

[0184] Compound 7

[0185] 1 1H NMR (400 MHz, DMSO-d6) δ 8.94 (d, J = 6.1 Hz, 1H), 8.71 (s, 1H), 8.66 (d, J = 5.2 Hz, 2H), 8.28–7.99 (m, 3H), 7.90 (dd, J = 8.9, 1.5 Hz, 1H), 7.63–7.44 (m, 2H), 4.67 (dd, J = 13.7, 7.1 Hz, 1H), 4.42 (t, J = 8.7 Hz, 1H), 3.62–3.50 (m, 1H), 3.44–3.31 (m, 1H), 2.84–2.66 (m, 1H), 2.38–2.06 (m, 1H).

[0186] Compound 8

[0187] 1 1H NMR (400 MHz, DMSO-d6) δ 9.10 (d, J = 24.5 Hz, 1H), 8.58 (d, J = 7.2 Hz, 1H), 8.33 (s, 1H), 8.11 (dd, J = 17.6, 8.7 Hz, 2H), 7.98 (ddd, J = 15.7, 12.8, 3.1 Hz, 4H), 7.52 (d, J = 8.3 Hz, 2H), 4.69 (dt, J = 13.5, 6.6 Hz, 1H), 4.47 (dt, J = 62.7, 8.9 Hz, 1H), 3.79 (s, 2H), 3.74–3.63 (m, 1H), 3.59–3.48 (m, 2H), 2.73 (dt, J = 13.0, 8.4 Hz, 1H), 2.28 (dt, J = 24.5, 9.8 Hz, 1H).

[0188] Compound 9

[0189] 1 H NMR (400MHz, DMSO-d6) δ10.24 (s, 1H), 9.64 (d, J = 1.9Hz, 1H), 9.22 (s, 1H), 9. 05(d,J=6.5Hz,1H),8.62(d,J=17.8Hz,2H),8.36–8.10(m,5H),8.09–7.93(m, 2H),7.84(t,J=7.5Hz,1H),4.69(dd,J=13.8,6.8Hz,1H),4.46(s,1H),3.55(d ,J=4.9Hz,1H),3.40(s,1H),2.74(dt,J=13.3,8.2Hz,1H),2.37–2.12(m,1H).

[0190] Compound 10

[0191] 1 H NMR(400MHz,MeOD)δ8.30(s,1H),7.85(dd,J=11.5,5.1Hz,2H),7.78(d,J=8.6Hz,3H) ,7.63(d,J=1.5Hz,1H),7.53(d,J=8.6Hz,2H),7.39(dd,J=8.9,2.0Hz,1H),4.77–4.41 (m,2H),3.74(dd,J=12.1,7.2Hz,1H),3.65–3.47(m,2H),2.89(dd,J=14.9,6.8Hz,1H) ,2.55–2.39(m,1H),1.86(d,J=7.0Hz,1H),1.69(dd,J=14.9,6.7Hz,1H),1.30(m,9H).

[0192] Compound 11

[0193] 1H NMR(400MHz,MeOD)δ8.41(d,J=11.5Hz,2H),8.01(d,J=8.9Hz,1H),7.88(dd ,J=15.4,6.4Hz,5H),7.73(s,1H),4.73(s,1H),4.53(t,J=8.4Hz,1H),3.75( dd,J=11.8,6.7Hz,1H),3.60(dd,J=7.6,5.3Hz,1H),2.91(s,1H),2.48(d,J =6.2Hz,1H),1.95–1.81(m,1H),1.69(dd,J=14.7,6.5Hz,1H),1.42(s,18H).

[0194] Compound 12

[0195] 1 H NMR(400MHz, DMSO-d6)δ9.18(s,1H),8.57(s,1H),8.21(s,1H),8.13–7.98(m,3H),7.92–7.80(m,1H),7.48–7.18(m,2H),7.00(d,J=8.2H z,1H),4.67(d,J=7.5Hz,1H),4.31(s,4H),4.13(s,1H),3.46(d,J=6.8Hz,3H),2.84–2.63(m,1H),2.28–2.14(m,1H),2.06–1.85(m,2H).

[0196] Compound 13

[0197] 11H NMR (400 MHz, DMSO-d6) δ 9.91 (s, 1H), 8.87 (d, J = 6.7 Hz, 1H), 8.50 (s, 1H), 8.23 (s, 1H), 8.12 (dd, J = 17.0, 8.7 Hz, 2H), 7.97 (d, J = 8.5 Hz, 1H), 7.91–7.84 (m, 1H), 7.64 (s, 1H), 7.57 (d, J = 7.9 Hz, 1H), 7.50 (t, J = 7.8 Hz, 1H), 7.29 (d, J = 7.9 Hz, 1H), 4.61 (dd, J = 13.3, 6.7 Hz, 1H), 3.98 (t, J = 8.3 Hz, 1H), 3.44 (dd, J = 11.5, 6.8 Hz, 2H), 3.29 (dd, J = 11.5, 6.2 Hz, 2H), 3.07 (s, 3H), 2.66–2.52 (m, 1H), 2.24–2.08 (m, 1H), 2.01 (dd, J = 15.1, 7.5 Hz, 1H).

[0198] Compound 14 [[ID=…]]

[0199] 1 1H NMR (400 MHz, DMSO-d6) δ 9.02 (d, J = 6.5 Hz, 1H), 7.78 (d, J = 8.3 Hz, 2H), 7.67 (t, J = 5.7 Hz, 3H), 7.56–7.38 (m, 3H), 4.33 (dd, J = 13.8, 6.9 Hz, 1H), 3.71 (t, J = 8.3 Hz, 1H), 3.30 (dd, J = 11.3, 6.7 Hz, 3H), 3.05 (dd, J = 11.4, 6.5 Hz, 1H), 2.46–2.35 (m, 1H), 2.01–1.71 (m, 1H), 1.35 (s, 9H).

[0200] Compound 15

[0201] 1 1H NMR (400 MHz, DMSO-d6) δ 8.64 (d, J = 8.7 Hz, 1H), 7.87 (d, J = 8.0 Hz, 2H), 7.66 (d, J = 7.9 Hz, 2H), 7.52–7.25 (m, 4H), 4.50 (dd, J = 15.0, 5.6 Hz, 2H), 3.76 (t, J = 6.4 Hz, 2H), 3.24–3.11 (m, 2H), 1.76 (dd, J = 35.4, 15.8 Hz, 4H), 1.57–1.12 (m, 7H).

[0202] Compound 16 (注:原文中“[[ID=…]]”部分你未给出完整准确内容,我按原样保留并翻译了其他部分。)

[0203] 1 H NMR (400MHz, DMSO-d6) δ8.55(s,1H),7.78(d,J=8.0Hz,2H),7.33(d,J=8.2Hz,2H),7.13(d,J=2.0Hz,1H),7.07(dd,J=8.4,1.8Hz,1H),6.88(d,J=8.5Hz ,1H),6.63(d,J=12.3Hz,1H),6.54(d,J=12.3Hz,1H),4.50(s,1H),3.79(s, 3H),3.45–3.17(m,3H),3.10–2.88(m,1H),2.16–1.83(m,1H),1.19(s,9H).

[0204] Compound 17

[0205] 1 H NMR (400MHz, DMSO-d6) δ8.47(d,J=6.6Hz,1H),7.69(dd,J=24.1,8.1Hz,5H),7.54–7.34(m,4H),6.66(d,J=15.9Hz,1H),4.40(dd ,J=13.9,7.0Hz,1H),3.76(t,J=8.2Hz,1H),3.36(d,J=6.8Hz,1H),3.06(dd,J=11.6,6.7Hz,1H),2.08–1.79(m,1H),1.35(s,9H).

[0206] Compound 18

[0207] 1 H NMR (400MHz, DMSO-d6) δ8.54(d,J=6.9Hz,1H),7.66(d,J=8.4Hz,2H),7.23(dd,J=8.5,2.3Hz,1H),7.18–7.07(m,3H),7.06–6.96(m,2H),4.47(q ,J=6.6Hz,1H),3.84(s,3H),3.74(t,J=8.1Hz,1H),3.13(dd,J=11.5,6.1Hz,1H),2.47–2.32(m,1H),1.98(dt,J=13.5,6.9Hz,1H),1.21(s,9H).

[0208] Compound 19

[0209] 1 H NMR (400MHz, DMSO-d6) δ8.67(d,J=6.9Hz,1H),7.86(q,J=8.3Hz,4H),7.73–7.53(m,2H),7.33(s,1H),7.08(d,J=8.6Hz,1H),4.52(q ,J=6.3Hz,1H),3.88(s,3H),3.77(t,J=7.9Hz,1H),3.20(dd,J=11.4,5.9Hz,2H),2.49–2.39(m,1H),2.14–1.95(m,1H),1.38(s,9H).

[0210] Biological experimental methods

[0211] The present invention is described below with reference to specific examples. It should be noted that these examples are merely illustrative and do not limit the present invention in any way.

[0212] Test Example 1: Application of amino acid compounds in an in vitro T cell differentiation experiment.

[0213] (1) Equipment preparation: surgical instruments (ophthalmic surgical scissors, ophthalmic forceps), pipette, 70 μM filter, 1 ml syringe, mouse fixation plate, LS column, magnetic stand, flow cytometry tube with cap, 70 μM flow cytometry tube with cap, 50 mL centrifuge tube, 15 mL centrifuge tube, 1.5 mL EP tube, 200 μL EP tube, flat-bottom 96-well plate, U-bottom 96-well plate.

[0214] (2) Reagent preparation: staining buffer (1xPBS + 2% super fine FBS), red blood cell lysis buffer, Miltenyi CD4 (L3T4) Microbeads, T cell culture medium (1640 + 10% super fine FBS + 1% penicillin-streptomycin solution + 0.1% β-mercaptoethanol), phosphoenol compounds to be tested (freeze at -80 °C with a concentration of 50 mM solvent DMSO), anti-CD3, anti-CD28, IL6 (50 ng / μL), IL23 (25 ng / μL), IL1β (10 ng / μL), TGFβ (2 ng / μL), IL-2 (40 ng / μL), IL12 (10 ng / μL), anti-IL4 (20 ng / μL), anti-IFN γ, MOG35-55, OVA257-264, GolgiStop, Ionomycin, PMA, cell membrane perforation solution, cell fixative, cell nucleus perforation solution, perforation solution washing solution (1xPerm), flow cytometry antibodies CD4, CD44, CD62L, CD25, Live / Dead, CD8, IL17A, IL17F, IFNγ, IL4, GM-CSF, RORγt, FoxP3, Phospho-STAT3, Phospho-STAT5.

[0215] (3) Use mice of special genotype for in vitro culture. Take 8-12 weeks old 2D2, OT-I genotype mice raised in an SPF animal breeding room. After euthanasia, use sterile surgical instruments in a cell clean bench to separate the mouse spleen, place it in a 70μM filter placed on a 50mL tube, use the handle of a 1mL syringe to fully grind the tissue, and use staining buffer to rinse all the cells on the filter into the 50mL tube, and centrifuge at 500G for 5 minutes. After discarding the supernatant, add 8mL / mouse red blood cell lysis buffer to the tube, resuspend the cells, let it stand at room temperature for 5 minutes, add an equal volume of staining buffer and mix to stop lysis; centrifuge at 500G to remove the supernatant. Resuspend the cells with 1mL staining buffer, filter the cells using a 70μM flow cytometry tube with a cap, centrifuge at 500G again to remove the supernatant, and resuspend the cells with T cell culture medium to obtain a whole spleen cell suspension of mice of special genotype.

[0216] (4) In vitro cell culture of whole spleen cells of mice with special genotypes. Count the cells of 1 mL of cell suspension, adjust the cell suspension concentration to 2×10^6 cells / mL, add cytokines for differentiation into Th17 or Tc17 (IL6 working concentration is 20 ng / mL, TGFβ working concentration is 1 ng / mL), cytokines for differentiation into Th2 (IL4 working concentration is 100 ng / mL, anti-IFNγ working concentration is 5 μg / mL), cytokines for differentiation into Th1 (IL2 working concentration is 20 ng / mL, IL12 working concentration is 10 ng / mL), cytokines for differentiation into Treg (TGFβ working concentration is 1 ng / mL), and add 50 μg / mL of peptide MOG35-55 (2D2 genotype mice) or 0.5 μg / mL of peptide OVA257-264 (OT-I genotype mice) and mix well. Obtain the cell fluid of the amino acid compound to be tested.

[0217] (5) When using C57BL / 6 wild-type mice for in vitro culture, it is necessary to coat the cell culture plates with antibodies in advance: dilute anti-CD3 and anti-CD28 in PBS at a working concentration of 5 μg / mL the night before the experiment, and add the dilution to a 96-well plate at 100 μL / well; seal the 96-well plate with sealing film and place it in a 4°C refrigerator, away from light, overnight.

[0218] (6) Childish Isolation of CD4-positive T cells. 8-12 week-old C57BL / 6 wild-type mice housed in an SPF animal care facility were euthanized. The spleen and peripheral lymph nodes were isolated using sterile surgical instruments in a cleanroom. The cells were then placed through a 70 μM filter placed on a 50 mL tube. The tissue was thoroughly triturated using the handle of a 1 mL syringe. All cells on the filter were rinsed into the 50 mL tube with staining buffer and centrifuged at 500 g for 5 minutes. The supernatant was discarded, and 8 mL / mouse of red blood cell lysis buffer was added to the tube to resuspend the cells. After incubation at room temperature for 5 minutes, an equal volume of staining buffer was added and mixed to terminate lysis. The supernatant was then centrifuged at 500 g and the supernatant discarded. The cells were resuspended in 0.5 mL of staining buffer and filtered through a 70 μM flow cytometer tube with a cap. 50 μL of CD4 (L3T4) Microbeads were then added and mixed. The tube was incubated at 4°C in the dark for 15 minutes. Add 4mL staining buffer to the flow tube, centrifuge at 500G and discard the supernatant, resuspend the cells with 1mL staining buffer / mouse, add to the LS column placed on the magnetic stand, wait for the liquid to drip clean, and then add staining buffer three times, 3mL / time. Remove the column, flush the cells into a 15mL tube with staining buffer, centrifuge and discard the supernatant. Dilute the flow cytometry antibodies CD4, CD44, CD62L and CD25 (500μL / mouse) with staining buffer at a ratio of 1:400, resuspend the cells with the antibody dilution solution, incubate at 4℃ in the dark for 30 minutes, and then wash the cells with staining buffer. After resuspending the cells with staining buffer, filter them into the tube using a 70μM flow tube with a cap, and finally sort the cells marked as CD4 positive, CD44 negative, CD62L positive and CD25 negative into the collection tube through a flow sorter to obtain naive cells. CD4-positive T cells.

[0219] (7) C57BL / 6 wild-type mice were selected from naive mice In vitro cell culture of CD4 positive T cells. CD4+ T cells were resuspended in T cell culture medium (concentration: 5 × 10^5 cells / mL), and cytokines for Th17 differentiation were added (IL6 working concentration: 20 ng / mL, TGFβ working concentration: 1 ng / mL, IL1β working concentration: 10 ng / mL, IL23 working concentration: 25 ng / mL), and mixed thoroughly. This gave the cell fluid to be mixed with the amino acid compounds to be tested.

[0220] (8) Divide the cell suspension mixed with the amino acid compound to be tested in the previous step into several 1.5mL EP tubes, 400μL / tube. Dilute the test compound to 100 times the working concentration with T cell culture medium, then add it to the cell suspension and mix thoroughly. 400μL of cell suspension requires 4μL of 100X compound diluent. For the control group, add the same volume of the amino acid compound to the corresponding T cell culture medium and mix thoroughly. Pipette the same volume of liquid as the 100X compound diluent and mix it with the cell suspension. After mixing the liquid in each 1.5mL EP tube, add it to the corresponding 96-well plate, 100μL / well. Place in a 37℃ cell culture incubator and culture for 48 hours or 96 hours.

[0221] (9) 48 hours (whole spleen cells of special genotype mice cultured in vitro) or 96 hours (selected naive spleen cells from C57BL / 6 wild-type mice) After in vitro cell culture of CD4-positive T cells), the cells were resuspended and transferred to a U-bottom 96-well plate, centrifuged at 500G to remove the supernatant, 100 μL T cell stimulation solution (GolgiStop, Ionomycin, PMA) was added to each well, and the cells were placed in a 37°C cell culture incubator for stimulation for 4 hours. The cell plate was removed and the cells were washed with staining buffer, centrifuged at 500G to remove the supernatant, and 50 μL surface antibody dilution solution was added to each well to perform flow cytometry staining on the cells (Live / Dead dilution ratio of 1:4000; according to the cell type, Th is CD25 and CD4; Tc is CD25 and CD8, dilution ratio 1:400), and placed in a 4°C refrigerator away from light for staining for 30 minutes. The cells were washed with buffer, centrifuged at 500G to remove the supernatant, fixed and punched with punching solution for 50 minutes or overnight, washed with 1xPerm, centrifuged at 800G to remove the supernatant, and 50 μL of cytokine (IL17A, IL17F, GM-CSF, IFNγ, IL4, dilution ratio 1:400) or transcription factor (RORγt, FoxP3, Phospho-STAT3, Phospho-STAT5 dilution ratio 1:200) antibody dilution solution was added to each well to stain the cells for 2 hours or overnight, and finally the experimental results were recorded by flow cytometry.

[0222] (10) The experimental results showed that the amino acid compound (administration group) could inhibit the secretion of cytokines IL17A, IL17F, and GM-CSF by Th17 cells (Figure 1-A). The experimental results further showed that the amino acid compound (administration group) could significantly inhibit the expression of RORγt (Figure 1-B) and significantly reduce the phosphorylation of the transcription factor STAT3 (Figure 1-C).

[0223] (11) The experimental results showed that amino acid compounds (administered group) could inhibit the secretion of cytokine IL4 by Th2 cells (Figure 1-D). Since IL4 plays an extremely important role in allergic diseases such as asthma, the results of this experiment revealed that amino acid compounds can slow the occurrence of allergic diseases such as asthma.

[0224] Test Example 2: Application of amino acid compounds in mouse autoimmune model.

[0225] (1) Supplies: Supplies required for in vitro cell culture are as described in Example 1, including 24-well plates, BD brand 1 mL syringes, and ordinary 1 mL syringes.

[0226] (2) Reagents: The reagents required for in vitro cell culture are as described in Example 1, including peptide MOG35-55, pertussis toxin, PBS, DMF, Tween 80, and anesthetic.

[0227] (3) The spleen cells of 2D2 genotype mice were induced to differentiate into Th17 in vitro (for detailed operation, see Test Example 1 above), and IL6, TGFβ, IL1β, IL23 and peptide MOG35-55 were added. The cell solution was added to a 24-well plate at 500 μL / well and placed in a 37°C cell culture incubator. After 34 hours, 20 μM of the amino acid compound to be tested was added and incubated for another 14 hours. The cells were washed out and injected into RAG1 gene knockout mice through the tail vein using a BD brand 1 mL syringe at a rate of 1.5x10^6 cells per recipient mouse, 300 μL / mouse. 24 hours later, the mice were intraperitoneally injected with pertussis toxin using a regular 1 mL syringe, 300 ng of the solvent was PBS / mouse, 200 μL / mouse.

[0228] (4) Administration to mice. PBS, DMF, and Tween80 were thoroughly mixed in a ratio of 88:10:2 and filtered through a 0.22 filter to obtain a PDT solvent. The amino acid compound to be tested was added to the PDT solvent at 250 μg / 200 μL; an equal amount of DMSO was added to the control group. The prepared drug was injected into the mouse via the peritoneal cavity at 200 μL / mouse. The drug was administered daily for 18 consecutive days. After 28 days, the popliteal lymph nodes of the mice were isolated and cell suspensions were prepared for flow cytometry analysis of Th17 cell-related characteristic molecules. 24 hours later, the amino acid compound to be tested was injected into the mouse peritoneal cavity using an ordinary 1 mL syringe at a dosage of 20 μL of drug per mouse per day (drug concentration of 50 mM) for 18 consecutive administrations.

[0229] (5) Starting from the return of cells through the tail vein, the weight of the mice was recorded every day and the clinical disease scores of the mice were observed. Clinical disease scores: healthy state (0 points); weak tail (1 point); slow walking, uncoordinated movement of the left and right hind limbs, and involuntary tremors of the hind limbs (2 points); paralysis of the hind limbs alone (3 points); paralysis of both hind limbs (4 points); paralysis of the forelimbs, or death of the mouse (5 points). On the 28th day after the return of cells through the tail vein, the central nervous system (CNS) of the mice was removed and a single cell suspension was prepared. The proportion and number of cells contained in the suspension were analyzed using flow cytometry.

[0230] (6) The experimental results showed that the disease incidence score of autoimmune encephalomyelitis (EAE) in the drug group was significantly lower than that in the control group (Figure 2-A), and the weight loss in the drug group was also significantly lower than that in the control group (Figure 2-B). This shows that amino acid compounds can significantly delay the progression of autoimmune diseases in mice.

Claims

1. A compound, which is a compound represented by formula (I) or a stereoisomer or a pharmaceutically acceptable salt of a compound represented by formula (I): in: n is 1, 2, or 3; R 1 For hydrogen, C 1-18 Alkylene, C 1-18 heteroalkylene, substituted or unsubstituted arylene, substituted or unsubstituted heteroarylene; R 2 For hydrogen, C 1-18 Alkylene, C 1-18 heteroalkylene, substituted or unsubstituted arylene, substituted or unsubstituted heteroarylene and -C(O)R 4 The substituents are composed of 4 C 1-18 Alkylene, C 1-18 heteroalkylene, substituted or unsubstituted arylene, substituted or unsubstituted heteroarylene; R 3 For hydrogen, C 1-18 Alkylene, C 1-18 Heteroalkylene, substituted or unsubstituted arylene, substituted or unsubstituted heteroarylene, OH, NH2, SH, N3, CN, COOH, SO3H, PO3H, alkyl or aryl carbonate, alkyl or aryl sulfonate, alkyl or aryl phosphate; L1 is selected from: single bond, C1-C6 alkylene, C1-C6 heteroalkylene, C 3-18 Cycloalkylene, C 3-18 Heterocyclylene, N(R 5 )、O、S、C(O)、N(R 5 )C(O), C(O)N(R 5 ), C(O)O, OC(O), SO, SO2, PO3, N(R 5 )SO2 or SO2N(R 5 ), where R 5 Selected from: hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, heterocycloalkyl, heterocycloalkylalkyl; Y1 is selected from: substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted alkenyl or substituted or unsubstituted alkynyl; L2 is selected from: single bond, C1-C6 alkyl, C1-C6 heteroalkyl, C3-C6 cycloalkyl, heterocyclic group, C2-C 24 Alkenyl, partially substituted or fully substituted C2-C 24 Alkenyl, substituted or unsubstituted alkynyl, N(R 5 )、O、S、C(O)、C(O)N(R 5 )、N(R 5 )C(O), C(O)O, OC(O), SO, SO2 and N(R 5 )SO2,R 5 Selected from: hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, heterocycloalkyl, heterocycloalkylalkyl; Y2 is selected from substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted alkenyl or substituted or unsubstituted alkynyl.

2. The compound according to claim 1, characterized in that The compound represented by formula (I) has at least one of the following structures: Demonstration compound skeleton: Optionally, the compound comprises a molecule selected from the group consisting of:

3. Use of the pharmaceutical composition in preparing a drug, characterized in that: The pharmaceutical composition is used for treating inflammatory diseases, and the pharmaceutical composition comprises the compound according to claim 1 or 2.

4. The use according to claim 3, characterized in that The compound regulates the metabolic level in cells, and the compound is used in an amount of 0.1 to 500 mg / kg.

5. The use according to claim 3 or 4, characterized in that The inflammatory disease is selected from autoimmune diseases and allergic diseases such as asthma.

6. The use according to claim 5, characterized in that The inflammatory disease is caused by differentiation of at least one selected from T cells, dendritic cells, eosinophils, neutrophils, macrophages, natural lymphocytes and monocytes.

7. The use according to claim 5, characterized in that The autoimmune disease is an immune response caused by self-antigens.

8. The use according to claim 5, characterized in that The asthma is induced by allergens.

9. The use according to claim 5, characterized in that The compound can be used alone or in combination with other compounds to treat inflammatory diseases.

10. A method for treating inflammatory diseases, characterized in that: The method comprises administering a pharmaceutically effective amount of the compound of claim 1 to a patient with an inflammatory disease.

11. Use of the compound of claim 1 in treating inflammatory diseases.

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