Oleoyl-ACP hydrolyase small molecule inhibitors and uses thereof

Abstract

The application discloses a kind of compound with oleoyl-ACP hydrolytic enzyme (OLAH) inhibitory activity, pharmaceutical composition and purposes thereof.The compound has the structure shown in formula A:Ar 1 -(L 1 ) a -X-(L 2 ) b -Ar 2 Wherein Ar 1 And Ar 2 Each is aryl, fused aryl, heteroaryl, cycloalkyl or heterocyclyl, L 1 And L 2 It is connecting group, and X is connecting group or heterocyclyl.The compound includes the compound shown in formula I~formula VII.Experiments show that the compound has OLAH inhibitory activity, and part of the compound can be directly combined with OLAH and inhibit respiratory syncytial virus replication.The application also relates to the purposes of the compound in preparing OLAH inhibitor, anti-respiratory syncytial virus drug or composition, and detecting OLAH activity or screening OLAH inhibitor kit.

Description

Technical Field

[0001] This invention belongs to the pharmaceutical field, specifically relating to small molecule inhibitors of oleoyl-ACP hydrolase and their uses. Background Technology

[0002] Respiratory syncytial virus (RSV) is a common single-stranded negative-sense RNA respiratory virus that primarily infects infants, the elderly, and immunocompromised individuals. It is a significant pathogen causing acute lower respiratory tract infections. RSV infection can lead to bronchiolitis, pneumonia, and in severe cases, respiratory failure and even death. Due to RSV's high transmissibility and the increased likelihood of severe illness in high-risk groups, the development of safe and effective anti-RSV drugs is of great importance.

[0003] Currently, interventions for RSV infection mainly include vaccination, monoclonal antibody prophylaxis, and antiviral drug therapy. However, existing antiviral drugs primarily target proteins within the virus itself, which may lead to issues such as decreased susceptibility due to viral mutations, the development of drug resistance, or limited applicability to certain populations. Therefore, developing anti-RSV drugs targeting host factors upon which viral replication depends is a promising research direction.

[0004] Oleoyl-acyl-carrier-protein hydrolase (OLAH) is an enzyme involved in fatty acid metabolism. Recent research and commentaries published in *Cell* (Cell, 2024, 187, 4586-4604; Cell, 2024, 187, 4549-4551) indicate that OLAH plays a crucial pathogenic role in severe respiratory viral infections. RSV infection induces upregulation of OLAH expression in host cells, promoting oleic acid production.

[0005] Further research revealed that OLAH-mediated oleic acid production is closely related to RSV infection progression. On the one hand, oleic acid can promote the accumulation of intracellular lipid droplets, creating a lipid-rich intracellular environment conducive to viral replication; on the other hand, oleic acid can affect the function of the antiviral protein IFITM3 and activate the NF-κB inflammatory signaling pathway, promoting the release of inflammatory factors. Furthermore, OLAH-related lipid metabolism disorders may also affect immune cell function, leading to an imbalance in the antiviral immune response.

[0006] Previous studies have shown that lipid droplet formation inhibitor ML-262 and fatty acid synthase inhibitor C75 can inhibit viral infection in wild-type macrophages; however, in OLAH-deficient cells, the antiviral effects of these inhibitors are significantly weakened or absent. These results suggest that OLAH-mediated rapid lipid droplet generation may be an important host cell event upon which efficient RSV replication depends, and OLAH could serve as a potential target for anti-RSV drug development.

[0007] However, there is still a lack of effective small molecule inhibitors for OLAH. It is necessary to develop small molecule compounds that can directly bind to and inhibit OLAH activity and have anti-RSV effects to enrich the anti-RSV drug development strategy targeting host lipid metabolism pathways. Summary of the Invention

[0008] The present invention aims to at least partially solve one of the technical problems in the related art.

[0009] Therefore, one objective of this invention is to provide a class of compounds with oleoyl-ACP hydrolase (OLAH) inhibitory activity. These compounds can inhibit OLAH enzyme activity, some of which can directly bind to the OLAH protein, and have the potential for application in inhibiting respiratory syncytial virus (RSV) replication.

[0010] To achieve the above objectives, the first aspect of this application provides a compound, said compound being a compound represented by Formula A, or a tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt, or prodrug of a compound represented by Formula A: Formula A: Ar 1 -(L 1 ) a -X-(L 2 ) b -Ar 2 ; Among them, Ar 1 and Ar 2 Each is independently selected from unsubstituted or R-substituted C6-C. 20 Aryl, C8-C 24 Dysmerized aryl, 5-14 membered heteroaryl, 5-14 membered dysmerized heteroaryl, C3-C 10 cycloalkyl, C3-C 10 Heterocyclic groups, or ring systems formed by the fusion or spirolysis of the aforementioned groups; L 1 and L 2 Each is independently selected from single bonds, C1-C6 alkylene groups, C2-C6 alkenyl groups, -O-, -S-, -NH-, -N(R)a )-, -C(=O)-, -C(=S)-, -SO2-, -N=N-, -O-(CH2) c -O-, or a divalent linker formed by a combination of the above groups; X is selected from the absence of, -O-, -S-, -NH-, -N(R b -, -C(=O)-, -C(=S)-, -SO2-, -N=N-, 5-7 membered heterocyclic groups containing 1-4 heteroatoms selected from N, O, and S, or fused heterocyclic groups containing 1-4 heteroatoms selected from N, O, and S; a and b are each independently 0, 1, 2, 3, or 4; c is 1, 2, 3, 4, 5, or 6; R a and R b Each R is independently H or C1-C6 alkyl; each R is independently selected from hydroxyl, amino, halogen, cyano, carboxyl, C1-C6 alkoxycarbonyl, carbamoyl, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkylthio, sulfonic acid, sulfonate, oxo, C1-C6 alkylsulfonyl, methylenedioxy, and -C(=O)NR. c R d Among them, R c and R d Each is independently H or C1-C6 alkyl; wherein the heteroatoms in the 5-14 membered heteroaryl, 5-7 membered heterocyclic and fused heterocyclic groups are each independently selected from N, O and S; wherein the pharmaceutically acceptable salts include alkali metal salts.

[0011] In some embodiments of the present invention, the Ar 1 and Ar 2 Each is independently selected from unsubstituted or R-substituted phenyl, naphthyl, biphenyl, benzothiazolyl, benzothiophene, benzofuranyl, indolyl, quinolinyl, isoquinolinyl, thiazolyl, thiadiazolyl, pyridyl, C3-C8 cycloalkyl or C3-C8 heterocyclic groups.

[0012] In some embodiments of the present invention, each of the R groups is independently selected from hydroxyl, amino, fluorine, bromine, cyano, carboxyl, methoxy, methylthio, methyl, ethyl, propyl, ethoxycarbonyl, carbamoyl, sulfonic acid, sodium sulfonate, oxo, methylenedioxy, or -C(=O)NH2.

[0013] In some embodiments of the present invention, the L 1 and L 2Each is independently selected from single bonds, -CH2-, -CH=CH-, -O-, -S-, -NH-, -C(=O)-, -C(=S)-, -SO2-, -N=N-, -OCH2CH2O-, -OCH2CH2CH2O-, or divalent linkages formed by sequentially connecting two or three of these linkages.

[0014] In some embodiments of the present invention, X is selected from non-existent, -O-, -NH-, -C(=S)-, -N=N-, 5-7 membered heterocyclic groups containing N and S, or fused heterocyclic groups containing N, S and oxo groups.

[0015] In some embodiments of the present invention, the compound is selected from any of the following classes of compounds: nitrogen- and / or sulfur-containing heterocyclic compounds; fused aromatic ring compounds; polyhydroxy aryl thioamide compounds; aromatic compounds containing carboxyl aryl ethers and polyether linkers; or azosulfonate compounds.

[0016] In some embodiments of the present invention, the compound is selected from compounds of formula I, II, III, IV, V, VI or VII, or their tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts or prodrugs: , , , , , , .

[0017] In some embodiments of the present invention, the compounds are selected from compounds I, II, III, IV, V, VI, and VII.

[0018] The inventors used the fluorescent substrate 4-methylumbelliferyl ketone heptanoate (4-MUH) to determine the OLAH inhibitory activity of the above compounds. Experimental results showed that compounds I-VII all exhibited OLAH enzyme activity inhibition, with half-maximal inhibitory concentrations (IC50) of [amount missing]. 50 The effective concentrations ranged from 3.096 to 33.722 μM. Among them, compounds II, III, IV, V, and VII exhibited strong OLAH inhibitory activity.

[0019] In some embodiments of the present invention, compound I has an IC50 effect on OLAH. 50 The concentration was 33.35 μM; the IC50 of compound II against OLAH was... 50 The concentration was 3.096 μM; the IC50 of compound III against OLAH was 3.096 μM. 50The concentration was 3.342 μM; the IC50 of compound IV against OLAH was... 50 The concentration was 3.682 μM; the IC50 of compound V against OLAH was... 50 The concentration was 3.246 μM; the IC50 of compound VI against OLAH was... 50 The concentration was 33.722 μM; the IC50 of compound VII against OLAH was... 50 It is 7.84 μM.

[0020] In some embodiments of the present invention, surface plasmon resonance (SPR) technology was used to detect the interaction between the compounds and OLAH. The results showed that compounds I-VII could directly bind to the OLAH protein; the equilibrium dissociation constant K between compound I and OLAH was [not specified in the original text]. D The concentration was 5.8 μM; the Kc of compound II and OLAH was... D The K2 of compound III and OLAH was 2.9 μM; D The Kc of compound IV was 16.8 μM; the Kc of OLAH was... D The Km of compound V is 9.9 μM; the Km of compound V with OLAH is 9.9 μM. D The Kc of compound VI is 0.78 μM; the Kc of compound VI and OLAH is 0.78 μM. D The Kc of compound VII is 2.4 μM; the Kc of OLAH is 2.4 μM. D It is 3.5 μM.

[0021] A second aspect of this application provides a pharmaceutical composition comprising the compound described in the first aspect of this application, or its tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts, or prodrugs; the pharmaceutical composition is used for the prevention and / or treatment of diseases associated with abnormal oleyl-ACP hydrolase (OLAH) activity.

[0022] In some embodiments of the present invention, the pharmaceutical composition includes one or more pharmaceutically acceptable carriers, excipients, and / or excipients.

[0023] In some embodiments of the present invention, the disease associated with abnormal OLAH activity is a viral infection.

[0024] In some embodiments of the present invention, the viral infection is RSV infection.

[0025] The third aspect of this application provides the use of the compounds described in the first aspect of this application, or the pharmaceutical compositions described in the second aspect of this application, in the preparation of OLAH inhibitors.

[0026] In some embodiments of the invention, the use is for in vitro use, not for therapeutic purposes. For example, the compound or pharmaceutical composition can be used to prepare a reagent for in vitro inhibition of OLAH enzyme activity.

[0027] The fourth aspect of this application provides the use of the compounds described in the first aspect of this application, or the pharmaceutical compositions described in the second aspect of this application, in the preparation of a medicament for the prevention and / or treatment of diseases associated with abnormal OLAH activity or expression levels.

[0028] In some embodiments of the present invention, the disease is a viral infection, such as RSV infection.

[0029] The fifth aspect of this application provides the use of the compounds described in the first aspect of this application, or the pharmaceutical compositions described in the second aspect of this application, in the preparation of a medicament or composition for inhibiting the replication of respiratory viruses.

[0030] In some embodiments of the present invention, the respiratory virus is RSV.

[0031] In some embodiments of the present invention, at a concentration of 3 μM, compound I showed an inhibition rate of 31.82% against RSV, and compound II showed an inhibition rate of 47.94% against RSV; at a concentration of 0.4 μM, compound III showed an inhibition rate of 30.70% against RSV, compound IV showed an inhibition rate of 100% against RSV, compound V showed an inhibition rate of 53.77% against RSV, and compound VI showed an inhibition rate of 40.14% against RSV.

[0032] The sixth aspect of this application provides the use of the compounds described in the first aspect of this application in the preparation of kits for detecting OLAH activity or screening OLAH inhibitors.

[0033] In some embodiments of the present invention, the kit includes the compound, OLAH or an enzyme preparation containing OLAH, a reaction buffer, and a detection substrate. In some specific embodiments, the detection substrate is 4-MUH. The kit can be used for non-therapeutic in vitro OLAH activity assays, OLAH inhibitory activity evaluations, or OLAH inhibitor screening.

[0034] The compounds shown in Formula A and their specific representative compounds provided by this invention possess OLAH inhibitory activity. Compounds I through VII can all inhibit OLAH enzyme activity; among them, compounds I through VII can directly bind to the OLAH protein, and compounds I through VI exhibit inhibitory effects on RSV replication in cellular anti-RSV experiments. Therefore, the compounds provided by this invention can serve as candidate compounds for OLAH inhibitors and can be used to develop in vitro reagents for inhibiting OLAH activity, OLAH detection or screening kits, and candidate drugs or pharmaceutical compositions for RSV infection intervention.

[0035] Additional aspects and advantages of this application will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of this application. Attached Figure Description

[0036] The above and / or additional aspects and advantages of the present invention will become apparent and readily understood from the description of the embodiments taken in conjunction with the following drawings, in which: Figure 1 This is a gel image of oleyl-ACP hydrolase (OLAH) obtained by SDS-PAGE detection and purification in Example 1 of the present invention.

[0037] Figure 2 This is a graph showing the results of OLAH-catalyzed hydrolysis of the fluorescent substrate 4-methylumbelliferone heptanoate in Example 1 of the present invention.

[0038] Figure 3 The IC50 of compound I in Example 2 of this invention inhibits OLAH enzyme activity. 50 Value result.

[0039] Figure 4 The IC50 of compound II in Example 2 of this invention inhibits OLAH enzyme activity. 50 Value result.

[0040] Figure 5 The IC50 of compound III in Example 2 of this invention inhibits OLAH enzyme activity. 50 Value result.

[0041] Figure 6 The IC50 of compound IV in Example 2 of this invention inhibits OLAH enzyme activity. 50 Value result.

[0042] Figure 7 The IC50 value of compound V in Example 2 of this invention inhibits OLAH enzyme activity. 50 Value result.

[0043] Figure 8 The IC50 value of compound VI in Example 2 of this invention inhibits OLAH enzyme activity. 50 Value result.

[0044] Figure 9 The IC50 value of compound VII in Example 2 of this invention inhibits OLAH enzyme activity. 50 Value result.

[0045] Figure 10 The binding affinity (K) result of the interaction between compound I and OLAH in Example 3 of this invention. D value).

[0046] Figure 11 The binding affinity (K) result of the interaction between compound II and OLAH in Example 3 of this invention. D value).

[0047] Figure 12 The binding affinity (K) result of the interaction between compound III and OLAH in Example 3 of this invention. D value).

[0048] Figure 13 The binding affinity (K) results for the interaction between compound IV and OLAH in Example 3 of this invention are shown. D value).

[0049] Figure 14 The binding affinity (K) result of the interaction between compound V and OLAH in Example 3 of this invention. D value).

[0050] Figure 15 The binding affinity (K) result of the interaction between compound VI and OLAH in Example 3 of this invention. D value).

[0051] Figure 16 The binding affinity (K) result of the interaction between compound VII and OLAH in Example 3 of this invention. D value). Detailed Implementation

[0052] The embodiments of this application are described in detail below. The embodiments described below are exemplary and are only used to explain this application, and should not be construed as limiting this application.

[0053] It should be noted that the terms "first," "second," etc., are used for descriptive purposes only and should not be construed as indicating or implying relative importance, or implicitly specifying the number of technical features indicated. Therefore, a feature specified with "first," "second," etc., may explicitly or implicitly include one or more of that feature. Furthermore, in the description of this application, unless otherwise stated, "multiple" means two or more.

[0054] The endpoints of the numerical ranges disclosed herein and any numerical values ​​are not limited to the precise range or value, but should be understood to include values ​​close to said range or value. Without contradiction, the endpoint values ​​of various numerical ranges, the endpoint values ​​of various ranges and individual point values, and individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered as specifically disclosed herein.

[0055] Unless otherwise defined, all technical and scientific terms used herein have the meanings commonly understood by one of ordinary skill in the art to which this application pertains. In this document, the terms "comprising" or "including" are open-ended expressions, meaning they include the specified content but do not exclude other content.

[0056] In this document, the terms “optionally,” “optionally,” or “optionally” generally mean that the event, feature, or condition described below may or may not occur, and the description includes both the occurrence of the event, feature, or condition and the absence of the event, feature, or condition.

[0057] In this paper, the term "oleoyl-acyl-carrier-protein hydrolase (OLAH)" refers to an enzyme with oleoyl-acyl-carrier protein hydrolase activity. OLAH is involved in host lipid metabolism processes. Previous studies have suggested that changes in OLAH expression and / or activity during RSV infection may be related to oleic acid production, lipid droplet accumulation, and viral replication. Therefore, OLAH can serve as a potential host target protein for screening and developing anti-RSV candidate compounds.

[0058] In this document, the term "respiratory syncytial virus (RSV)" refers to a virus that can cause respiratory infections. RSV infection includes, but is not limited to, acute respiratory infections, bronchiolitis, pneumonia, and other RSV-related diseases or conditions caused by RSV.

[0059] In this document, the term "OLAH inhibitor" refers to a compound, composition, or formulation that can reduce OLAH enzyme activity, decrease OLAH-substrate interaction, and / or directly bind to OLAH protein in in vitro enzymatic assays, cell assays, or other suitable detection systems.

[0060] In this paper, the term "inhibition of RSV replication" refers to a reduction in viral plaque number, viral titer, viral nucleic acid level, viral protein expression level, and / or other indicators reflecting viral replication in virus-infected cells compared to a viral control without administration of the test compound. This reduction can be determined by plaque formation assays, cytopathic effect assays, immunostaining assays, nucleic acid quantification assays, or other suitable methods.

[0061] In this document, the term "pharmaceutically acceptable salt" means a salt that is suitable for contact with human or animal tissues within the bounds of reasonable medical judgment, has acceptable toxicity, irritation, and allergic risk, and has a reasonable benefit / risk ratio. Pharmaceutically acceptable salts include salts formed from inorganic acids, organic acids, inorganic bases, or organic bases.

[0062] According to embodiments of this application, the inorganic acids include, but are not limited to, hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, and nitric acid; the organic acids include, but are not limited to, formic acid, acetic acid, propionic acid, lactic acid, citric acid, tartaric acid, fumaric acid, maleic acid, succinic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, and trifluoromethanesulfonic acid. The inorganic bases include, but are not limited to, sodium hydroxide, potassium hydroxide, lithium hydroxide, calcium hydroxide, and magnesium hydroxide; the organic bases include, but are not limited to, ammonia, ethylamine, diethylamine, triethylamine, ethanolamine, diethanolamine, triethanolamine, tromethamine, piperazine, and piperidine.

[0063] In this document, the term "stereoisomer" refers to isomers formed due to differences in the spatial arrangement of atoms or groups in a molecule, including but not limited to enantiomers, diastereomers, cis-trans isomers, and conformational isomers. This application includes various stereoisomers and mixtures thereof of compounds shown in structural formulas I, II, III, IV, V, VI, and VII.

[0064] In this document, the term "tautomer" refers to a functional group isomer that can interconvert due to the migration of atoms, particularly hydrogen atoms, between different positions in a molecule. This application includes tautomer forms of compounds represented by structural formulas I-VII and mixtures thereof.

[0065] In this document, the term "solvent" refers to a compound formed by the combination of the present application compound or its salt with one or more solvents through intermolecular non-covalent interactions, wherein the solvents may be stoichiometric or non-stoichiometric. The solvents include, but are not limited to, water, methanol, ethanol, acetic acid, acetone, ethyl acetate, dichloromethane, and dimethyl sulfoxide. When the solvent is water, the solvate is a hydrate.

[0066] In this invention, "hydrate" is a special case of solvate, referring to a substance in which the solvent molecule is water.

[0067] In this document, the term "prodrug" refers to a derivative that can be converted into a biologically active compound of this application under physiological conditions or through processes such as solvation, enzymatic hydrolysis, or metabolism. The prodrug can be prepared by conventional modification of the modifiable functional groups in the compound and converted into the corresponding active compound in vivo.

[0068] In this document, the term "therapeuticly effective amount" refers to the amount of the compound or pharmaceutical composition of this application sufficient to produce the effect required to prevent, alleviate, improve, or treat a disease, condition, or symptom. Therapeuticly effective amount can be determined based on the subject's age, weight, route of administration, disease severity, treatment regimen, concomitant medications, and clinician judgment.

[0069] In this document, the term "pharmaceutical composition" means a composition comprising the compound described in this application, or a pharmaceutically acceptable salt, solvate, hydrate, or prodrug thereof, and a pharmaceutically acceptable carrier, excipient, and / or excipient.

[0070] In this document, the term "pharmaceutically acceptable carrier" includes carriers, diluents, fillers, absorption enhancers, adsorbents, binders, lubricants, humectants, buffers, flavoring agents, and sweeteners commonly used in the pharmaceutical field. Non-limiting examples include lactose, sucrose, mannitol, sorbitol, corn starch, cellulose and its derivatives, calcium phosphate, polyvinylpyrrolidone, polyethylene glycol, polyvinyl alcohol, stearic acid, magnesium stearate, calcium stearate, vegetable oils, and pharmaceutically acceptable surfactants.

[0071] This application discloses a class of small molecule compounds with oleoyl-acyl-carrier-protein hydrolase (OLAH) inhibitory activity, including compound I, compound II, compound III, compound IV, compound V, compound VI, and compound VII; and provides the use of said compounds as OLAH inhibitors, in the preparation of medicaments or compositions for inhibiting respiratory syncytial virus (RSV) replication, in pharmaceutical compositions for the prevention and / or treatment of RSV infection-related diseases, and in the preparation of reagents or kits for inhibiting OLAH activity, which will be described in detail below.

[0072] compound In a first aspect, this application provides a compound, said compound being a compound of formula A, or a tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt, or prodrug of a compound of formula A: Formula A: Ar 1 -(L 1 ) a -X-(L 2 ) b -Ar 2 ; Among them, Ar 1 and Ar 2 Each is independently selected from unsubstituted or R-substituted C6-C. 20 Aryl, C8-C 24 Dysmerized aryl, 5-14 membered heteroaryl, 5-14 membered dysmerized heteroaryl, C3-C 10 cycloalkyl, C3-C 10 Heterocyclic groups, or ring systems formed by the fusion or spirolysis of the aforementioned groups; L 1 and L2 Each is independently selected from single bonds, C1-C6 alkylene groups, C2-C6 alkenyl groups, -O-, -S-, -NH-, -N(R) a -, -C(=O)-, -C(=S)-, -SO2-, -N=N-, -O-(CH2)cO-, or divalent linkers formed by combinations of the above groups; X is selected from non-existent, -O-, -S-, -NH-, -N(R b -, -C(=O)-, -C(=S)-, -SO2-, -N=N-, 5-7 membered heterocyclic groups containing 1-4 heteroatoms selected from N, O, and S, or fused heterocyclic groups containing 1-4 heteroatoms selected from N, O, and S; a and b are each independently 0, 1, 2, 3, or 4; c is 1, 2, 3, 4, 5, or 6; R a and R b Each R is independently H or C1-C6 alkyl; each R is independently selected from hydroxyl, amino, halogen, cyano, carboxyl, C1-C6 alkoxycarbonyl, carbamoyl, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkylthio, sulfonic acid, sulfonate, oxo, C1-C6 alkylsulfonyl, methylenedioxy, and -C(=O)NR. c R d Among them, R c and R d Each is independently H or C1-C6 alkyl; wherein the heteroatoms in the 5-14 membered heteroaryl, 5-7 membered heterocyclic and fused heterocyclic groups are each independently selected from N, O and S; wherein the pharmaceutically acceptable salts include alkali metal salts.

[0073] According to embodiments of this application, the pharmaceutically acceptable salt is a sodium salt.

[0074] According to embodiments of this application, the Ar 1 and Ar 2 Each is independently selected from unsubstituted or R-substituted phenyl, naphthyl, biphenyl, benzothiazolyl, benzothiophene, benzofuranyl, indolyl, quinolinyl, isoquinolinyl, thiazolyl, thiadiazolyl, pyridyl, C3-C8 cycloalkyl or C3-C8 heterocyclic groups.

[0075] According to embodiments of this application, each of the R groups is independently selected from hydroxyl, amino, fluorine, bromine, cyano, carboxyl, methoxy, methylthio, methyl, ethyl, propyl, ethoxycarbonyl, carbamoyl, sulfonic acid, sodium sulfonate, oxo, methylenedioxy, or -C(=O)NH2.

[0076] According to an embodiment of this application, the L 1 and L 2Each is independently selected from single bonds, -CH2-, -CH=CH-, -O-, -S-, -NH-, -C(=O)-, -C(=S)-, -SO2-, -N=N-, -OCH2CH2O-, -OCH2CH2CH2O-, or divalent linkages formed by sequentially connecting two or three of these linkages.

[0077] According to embodiments of this application, X is selected from non-existent, -O-, -NH-, -C(=S)-, -N=N-, 5-7 membered heterocyclic groups containing N and S, or fused heterocyclic groups containing N, S and oxo groups.

[0078] According to embodiments of this application, the compound is selected from any of the following classes of compounds: nitrogen- and / or sulfur-containing heterocyclic compounds; fused aromatic ring compounds; polyhydroxy aryl thioamide compounds; aromatic compounds containing carboxyl aryl ethers and polyether linkers; or azosulfonate compounds.

[0079] According to embodiments of this application, the compound is selected from compounds represented by Formula I, II, III, IV, V, VI, or VII, or their tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts, or prodrugs: , , , , , , .

[0080] According to embodiments of this application, the compounds are selected from compounds I, II, III, IV, V, VI, and VII.

[0081] The inventors used the fluorescent substrate 4-methylumbelliferyl ketone heptanoate (4-MUH) to determine the OLAH inhibitory activity of the above compounds. Experimental results showed that compounds I-VII all exhibited OLAH enzyme activity inhibition, with half-maximal inhibitory concentrations (IC50) of [amount missing]. 50 The effective concentrations ranged from 3.096 to 33.722 μM. Among them, compounds II, III, IV, V, and VII exhibited strong OLAH inhibitory activity.

[0082] According to embodiments of this application, compound I has an IC effect on OLAH. 50 The concentration was 33.35 μM; the IC50 of compound II against OLAH was... 50 The concentration was 3.096 μM; the IC50 of compound III against OLAH was 3.096 μM. 50The concentration was 3.342 μM; the IC50 of compound IV against OLAH was... 50 The concentration was 3.682 μM; the IC50 of compound V against OLAH was... 50 The concentration was 3.246 μM; the IC50 of compound VI against OLAH was... 50 The concentration was 33.722 μM; the IC50 of compound VII against OLAH was... 50 It is 7.84 μM.

[0083] According to embodiments of this application, surface plasmon resonance (SPR) technology was used to detect the interaction between the compounds and OLAH. The results showed that compounds I-VII can directly bind to the OLAH protein; the equilibrium dissociation constant K between compound I and OLAH was... D The concentration was 5.8 μM; the Kc of compound II and OLAH was... D The K2 of compound III and OLAH was 2.9 μM; D The Kc of compound IV was 16.8 μM; the Kc of OLAH was... D The Km of compound V is 9.9 μM; the Km of compound V with OLAH is 9.9 μM. D The Kc of compound VI is 0.78 μM; the Kc of compound VI and OLAH is 0.78 μM. D The Kc of compound VII is 2.4 μM; the Kc of OLAH is 2.4 μM. D It is 3.5 μM.

[0084] The above results indicate that the compounds shown in formulas I to VII have inhibitory effects on OLAH enzyme activity. Among them, compounds II, III, IV, V, and VII exhibit strong OLAH inhibitory activity; compounds I to VII have experimental support for direct binding to OLAH.

[0085] According to embodiments of this application, the compound can be prepared by chemical synthesis or obtained by separation and purification methods. The specific synthetic route, reaction conditions, separation and purification methods, and structural identification methods can be determined based on the actual source of the compound, its structural characteristics, and conventional techniques in the art. For example, column chromatography, recrystallization, preparative high-performance liquid chromatography, or combinations thereof can be used for purification; nuclear magnetic resonance, mass spectrometry, infrared spectroscopy, elemental analysis, and / or high-performance liquid chromatography can be used for structural confirmation and purity detection.

[0086] According to embodiments of this application, the compound exists in free form, as a solvate, as a hydrate, as a pharmaceutically acceptable salt, or as a prodrug. For compounds containing acidic groups, alkali metal salts, such as sodium salts, may be formed; for compounds containing basic groups, addition salts with pharmaceutically acceptable acids may be formed.

[0087] Pharmaceutical Composition In a second aspect of this application, a pharmaceutical composition is provided. According to embodiments of this application, the pharmaceutical composition comprises the compound described in the first aspect of this application, or its tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts, or prodrugs; the pharmaceutical composition is used for the prevention and / or treatment of diseases associated with abnormal oleyl-ACP hydrolase (OLAH) activity.

[0088] According to embodiments of this application, the pharmaceutical composition includes one or more pharmaceutically acceptable carriers, excipients, and / or excipients.

[0089] According to an embodiment of this application, the disease associated with abnormal OLAH activity is a viral infection.

[0090] According to an embodiment of this application, the viral infection is an RSV infection.

[0091] According to embodiments of this application, the pharmaceutical composition is used for the prevention and / or treatment of diseases associated with abnormal OLAH activity and / or expression. In some embodiments, the disease is RSV infection or an RSV infection-related disease.

[0092] According to embodiments of this application, the pharmaceutical composition can be prepared into a formulation suitable for oral administration, including but not limited to tablets, capsules, granules, powders, oral liquids, suspensions, or emulsions.

[0093] According to embodiments of this application, the pharmaceutical composition can be prepared as a formulation suitable for parenteral administration, including but not limited to injections, lyophilized powder injections, sterile powders for injection, liposome formulations, or nanoformulations.

[0094] According to embodiments of this application, the pharmaceutical composition can be prepared as a formulation suitable for local administration to the respiratory tract, including but not limited to inhalers, nebulized inhalation formulations, sprays, nasal formulations, aerosols, dry powder inhalers, or nasal drops.

[0095] According to embodiments of this application, the dosage, frequency, and duration of administration of the compounds in the pharmaceutical composition can be determined based on the subject's age, weight, disease type, severity of infection, route of administration, dosage form, concomitant medications, and the judgment of the clinician.

[0096] use The third aspect of this application provides the use of the compound described in the first aspect of this application, or its tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, or the pharmaceutical compositions described in the second aspect of this application, in the preparation of OLAH inhibitors.

[0097] According to embodiments of this application, the use is for in vitro use without therapeutic purposes. For example, the compound or pharmaceutical composition can be used to prepare reagents for in vitro inhibition of OLAH enzyme activity, for evaluation of OLAH enzyme activity, evaluation of OLAH inhibitory activity, or related basic research.

[0098] According to embodiments of this application, the compound is used as an OLAH inhibitor in a fluorescent enzyme activity detection system using 4-MUH as a substrate.

[0099] use In a fourth aspect of this application, this application provides the use of the compound described in the first aspect of this application, or its tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, or the pharmaceutical compositions described in the second aspect of this application, in the preparation of a medicament for the prevention and / or treatment of diseases associated with abnormal OLAH activity and / or expression.

[0100] According to embodiments of this application, the disease associated with abnormal oleyl-ACP hydrolase (OLAH) activity or expression levels is a viral infection.

[0101] According to embodiments of this application, the disease associated with abnormal OLAH activity and / or expression is respiratory syncytial virus (RSV) infection. The RSV infection includes acute respiratory infections, bronchiolitis, pneumonia, or other RSV infection-related diseases caused by RSV.

[0102] According to embodiments of this application, the compound or pharmaceutical composition may be administered alone or in combination with one or more other treatment modalities. These other treatment modalities may include antiviral therapy, immunotherapy, anti-inflammatory therapy, oxygen therapy, respiratory support therapy, and / or symptomatic supportive therapy. Combined administration may be simultaneous, sequential, or administered at appropriate time intervals.

[0103] use In a fifth aspect of this application, this application provides the use of the compound described in the first aspect of this application, or its tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, or the pharmaceutical compositions described in the second aspect of this application, in the preparation of a medicament or composition for inhibiting the replication of respiratory viruses.

[0104] According to an embodiment of this application, the respiratory virus is respiratory syncytial virus.

[0105] According to embodiments of this application, the inhibitory effect of the compounds on RSV replication was evaluated using a plaque formation assay. At a concentration of 3 μM, compound I showed an inhibition rate of 31.82% on RSV, and compound II showed an inhibition rate of 47.94% on RSV; at a concentration of 0.4 μM, compound III showed an inhibition rate of 30.70% on RSV, compound IV showed an inhibition rate of 100% on RSV, compound V showed an inhibition rate of 53.77% on RSV, and compound VI showed an inhibition rate of 40.14% on RSV.

[0106] It should be understood that, based on the currently available data from cell anti-RSV assays, compounds I-VI have experimental support for inhibiting RSV replication in vitro.

[0107] use In a sixth aspect of this application, this application provides the use of the compound described in the first aspect of this application, or its tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, in the preparation of a kit for detecting oleyl-ACP hydrolase activity or screening oleyl-ACP hydrolase inhibitors.

[0108] According to embodiments of this application, the kit includes the compound, oleyl-ACP hydrolase or an enzyme preparation containing oleyl-ACP hydrolase, a reaction buffer, and a detection substrate. The kit may also include instructions for use, indicating the use of the compound for an in vitro OLAH enzyme activity inhibition assay, or as a positive control for an OLAH inhibitor screening assay.

[0109] The present invention will be further described below with reference to embodiments. Those skilled in the art should understand that the following embodiments are for illustrative purposes only and should not be considered as limiting the scope of protection of the present invention. Where specific techniques or conditions are not specified in the embodiments, they are performed according to conventional techniques in the art or the techniques or conditions described in the product manual. Reagents or instruments used, unless otherwise specified, are all conventional products that can be obtained through commercial channels.

[0110] Example 1: Expression, isolation, purification, and SDS-PAGE verification of OLAH protein 1. Expression, isolation, and purification of OLAH protein Under aseptic conditions, strains containing the OLAH recombinant expression vector (construction and management of recombinant strains are described in the literature) were... J. Biol. Chem. , 2016, 291 (7) 3520-3530, bacterial concentration of OD 600= 0.6) was inoculated at a 1:100 ratio into LB medium containing kanamycin (concentration 1 μL / mL) and cultured with shaking at 37 ℃ and 220 rpm for 12 h. After obtaining single colonies by streaking isolation, a single colony was picked and inoculated into 500 mL of LB medium containing kanamycin and cultured at 37 ℃ and 220 rpm for 12 h. Subsequently, isopropyl-β-D-thiogalactoside (IPTG) was added to induce expression for 4 h.

[0111] After culturing, the bacterial cells were collected by centrifugation and resuspended in PBS. The resuspended bacterial cells were then sonicated to disrupt the cytotoxicity. The sonication parameters were: 4 s sonication, 16 s interval, 5 min total sonication time, and 75% power. After sonication, the cells were centrifuged at 4 ℃ and 10,000 rpm for 15 min, and the supernatant was collected.

[0112] The supernatant was loaded onto a Ni-NTA nickel column pre-equilibrated with 10 mM imidazole at a flow rate of 1 mL / min. The throughflow was collected, and gradient elution was performed using 10–500 mM imidazole. The eluted fraction containing OLAH protein was collected and subjected to buffer replacement by gel size exclusion chromatography (SEC) to obtain OLAH protein in the target buffer. The target buffer composition was: 100 mM Tris, 50 mM NaCl, and 1 mM EDTA, pH 7.4.

[0113] 2. SDS-PAGE verification The purified OLAH protein sample was mixed with 5× protein loading buffer at a ratio of 4:1 (v / v) and heated at 100℃ for 10 min to denature the protein. After centrifugation, the supernatant was collected for SDS-PAGE electrophoresis. The electrophoresis conditions were 140 V constant voltage electrophoresis for 1 h.

[0114] After electrophoresis, the protein bands were stained with Coomassie Brilliant Blue for 1 hour and then destained for 2 hours to verify the expression and purification of OLAH protein.

[0115] The SDS-PAGE gel image of the purified OLAH is shown below. Figure 1 As shown.

[0116] 3. Verification of the ability to catalyze the hydrolysis of the fluorescent substrate 4-methylumbelliferone heptaester To evaluate the catalytic function of the purified OLAH protein, a fluorescence kinetic detection system was established using 4-methylumbelliferyl ketoheptanate (4-MUH) as a substrate. Before the experiment, the purified OLAH protein was adjusted to a working stock solution of 250 nM with dilution buffer (containing 100 mM Tris, 50 mM NaCl, and 1 mM EDTA, pH 7.4). The substrate 4-MUH and its product 4-methyl-7-hydroxycoumarin (4-MU) were dissolved in DMSO, with a stock solution concentration of 12 mM. Simultaneously, denatured and inactivated enzyme samples were prepared by heat treatment at 100 °C for 10 min, serving as an activity-dependent control.

[0117] Enzyme activity assays were performed in black 96-well plates: 1 μL of substrate stock solution was added to 99 μL of pre-prepared active enzyme solution or heat-inactivated enzyme solution to trigger the reaction. For the blank control, buffer solution was used instead of enzyme solution, and 1 μL of 4-MUH or 4-MU stock solution was added, respectively. The excitation / emission wavelengths were set to 350 / 450 nm, and continuous monitoring was performed for 60 min in top-reading mode, with a data acquisition interval of 5 min.

[0118] like Figure 2 As shown, the native conformation of OLAH exhibits significant substrate hydrolytic activity, with the fluorescence signal increasing throughout the reaction and approaching the theoretical fluorescence intensity of an equimolar concentration of the 4-MU standard at the endpoint. In contrast, the heat-denatured group and the enzyme-free blank group maintained baseline substrate background signals throughout the process. These results indicate that OLAH-mediated hydrolysis of 4-MUH depends on the active conformation of OLAH.

[0119] Example 2: Determination of the inhibitory effect of compounds I-VII on OLAH enzyme activity The inhibitory effects of compounds I–VII on the catalytic activity of OLAH were determined using the fluorescent substrate 4-methylumbelliferyl ketone heptanoate (4-MUH), and the half-maximal inhibitory concentration (IC50) was calculated. 50 ).

[0120] The SEC-purified OLAH protein was replaced with a buffer of 100 mM Tris, 50 mM NaCl, and 1 mM EDTA at pH 7.4, and the protein concentration was adjusted to 250 nM. 4-MUH and compounds I-VII were dissolved in DMSO to prepare stock solutions; the 4-MUH stock solution concentration was 12.1 mM, and the stock solutions concentrations of each compound were 10.1 mM. The stock solutions of each compound were then serially diluted.

[0121] In a 96-well black plate, 99 μL of OLAH protein solution and 1 μL of different concentrations of the test compound solution were added to each well, and incubated at 37 °C for 30 min. Subsequently, 1 μL of 4-MUH stock solution was added to each well to initiate the reaction. In the control wells, an equal volume of DMSO was used to replace the test compound.

[0122] Fluorescence signals were detected in top-reading mode under excitation wavelength of 350 nm and emission wavelength of 450 nm for 30 min, with continuous shaking and mixing during the detection period. The inhibition rate of each compound on OLAH enzyme activity was calculated based on the fluorescence signals, and the IC50 was further calculated. 50 value.

[0123] The inhibitory effects of compounds I-VII on OLAH enzyme activity are shown in Table 1. Figures 3-9 As shown.

[0124] Table 1: IC50 of compounds I-VII on OLAH enzyme activity 50 value

[0125] Example 3: Determination of the binding affinity of compounds I-VII to OLAH protein The binding affinity between compounds I-VII and OLAH protein was determined using surface plasmon resonance (SPR) technology.

[0126] Detection was performed at 25 °C using the Biacore™ 8K+ system and CM5 sensor chip. First, 200 μM 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) and 50 μM... N An activation solution was prepared by mixing hydroxysuccinimide (NHS) and injected into the CM5 chip surface at a flow rate of 10 μL / min for 420 s. Subsequently, the purified OLAH protein was dissolved in 10 mM sodium acetate buffer (pH 4.5) and injected into the detection channel at a flow rate of 10 μL / min for 600 s to immobilize the OLAH protein on the chip surface. The chip surface was then blocked with 1 M ethanolamine for 420 s.

[0127] The reference channel underwent the same activation and blocking procedures, but PBS buffer was injected instead of OLAH protein solution. After chip equilibration, compounds I–VII were prepared in concentration gradients ranging from 0.78 μM to 50 μM, and detection was performed using a multi-cycle kinetic mode. Each concentration of compound was injected at a flow rate of 30 μL / min, with a binding time of 120 s and a dissociation time of 300 s.

[0128] Using Biacore Insight Evaluation software, the sensor map was fitted based on a 1:1 Langmuir binding model to calculate the kinetic parameters and dissociation constant (K) between the compound and the OLAH protein. D ).

[0129] The results showed that compounds I-VII could directly bind to the OLAH protein, and their K D The values ​​ranged from 0.78 to 16.8 μM. The SPR affinity test results for compounds I–VII with OLAH protein are shown below. Figures 10-16 .

[0130] Example 4: Determination of the inhibitory activity of compounds I-VI against respiratory syncytial virus The inhibitory effects of compounds I-VI on respiratory syncytial virus (RSV) replication were determined using a plaque formation assay.

[0131] Hep-2 cells were stored at a density of 2-3 × 10⁶ cells per well. 4 Cells were seeded at a density of 100 μL in 96-well plates, with 100 μL of DMEM medium added to each well, and incubated overnight. The RSV laboratory-adapted strain was serially diluted 10-fold with pre-cooled serum-free DMEM medium, and the diluted virus solution was stored on ice for later use.

[0132] The following day, the cell culture supernatant was discarded, and each well was washed once with PBS. Then, 100 μL of virus solution at different dilutions was added to each well, and the cells were incubated at 37 ℃ for 1–2 h for adsorption. Virus-free control wells and virus control wells were set up; serum-free DMEM was added to the virus-free control wells, while virus solution was added to the virus control wells without the test compound.

[0133] After virus adsorption, 1 μL of solution of compounds I-VI prepared in serum-free DMEM was added to each well; subsequently, 100 μL of 1% methylcellulose covering medium preheated to 37 °C was added to each well. The final concentration of the test compounds was 3 μM or 0.4 μM, with two replicates per group. Cells were incubated at 37 °C in a 5% CO2 incubator for 3 days.

[0134] After incubation, discard the culture medium and add 100 μL of pre-chilled acetone-methanol fixative (acetone to methanol volume ratio 60:40) to each well. Fix at room temperature for 20 min. After fixation, air dry naturally. Add 20 μL of PBST (phosphate buffer containing Tween 20) to each well, gently shake to wash, and discard. Then add 100 μL of blocking buffer and block at room temperature for 30–60 min.

[0135] Subsequently, anti-RSV primary antibody (commercially available) diluted with blocking buffer was added to each well to a final concentration of 2 μg / mL, and incubated at room temperature for 1 h. After washing three times with PBST, alkaline phosphatase-labeled goat anti-mouse IgG (H+L) secondary antibody (commercially available) was added, and incubated for 1 h. After washing three times again with PBST, 50–100 μL of BCIP / NBT alkaline phosphatase chromogenic solution was added for color development. The reaction was terminated with water after color development was complete. The inhibition rate of the compound against RSV was calculated by plaque counting.

[0136] The inhibition results of compounds I and II on RSV at a concentration of 3 μM are shown in Table 2; the inhibition results of compounds III to VI on RSV at a concentration of 0.4 μM are shown in Table 3.

[0137] Table 2: Inhibition rate of compounds I-II on RSV

[0138] Table 3: Inhibition rate of compounds III-VII on RSV

[0139] In the description of this specification, the references to "one embodiment," "some embodiments," "example," "specific example," or "some examples," etc., indicate that a specific feature, structure, material, or characteristic described in connection with that embodiment or example is included in at least one embodiment or example of this application. In this specification, the illustrative expressions of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples. Moreover, without contradiction, those skilled in the art can combine and integrate the different embodiments or examples described in this specification, as well as the features of different embodiments or examples.

[0140] Although embodiments of this application have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting this application. Those skilled in the art can make changes, modifications, substitutions and variations to the above embodiments within the scope of this application.

Claims

1. A compound, characterized in that, The compound is the compound represented by Formula A, or a tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt, or prodrug of the compound represented by Formula A: Formula A: Ar 1 -(L 1 ) a -X-(L 2 ) b -Ar 2 ; in, Ar 1 and Ar 2 Each is independently selected from unsubstituted or R-substituted C6-C. 20 Aryl, C8-C 24 Fused aryl, 5-14 fused heteroaryl, 5-14 fused heteroaryl, C3-C 10 cycloalkyl, C3-C 10 Heterocyclic groups, or ring systems formed by the fusion or spirofusion of the aforementioned groups; L 1 and L 2 Each is independently selected from single bonds, C1-C6 alkylene groups, C2-C6 alkenyl groups, -O-, -S-, -NH-, -N(R) a )-, -C(=O)-, -C(=S)-, -SO2-, -N=N-, -O-(CH2) c -O-, or a divalent linker formed by a combination of the above groups; X is selected from non-existent, -O-, -S-, -NH-, -N(R) b -, -C(=O)-, -C(=S)-, -SO2-, -N=N-, 5-7 membered heterocyclic groups containing 1 to 4 heteroatoms selected from N, O and S, or fused heterocyclic groups containing 1 to 4 heteroatoms selected from N, O and S; a and b are each independently 0, 1, 2, 3 or 4; c is 1, 2, 3, 4, 5, or 6; R a and R b Each is independently H or C1-C6 alkyl; Each R is independently selected from hydroxyl, amino, halogen, cyano, carboxyl, C1-C6 alkoxycarbonyl, carbamoyl, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkylthio, sulfonic acid, sulfonate, oxo, C1-C6 alkylsulfonyl, methylenedioxy, and -C(=O)NR. c R d ; Among them, R c and R d Each is independently H or C1-C6 alkyl; In this context, the heteroatoms in the 5-14 membered heteroaryl groups, 5-7 membered heterocyclic groups, and fused heterocyclic groups are each independently selected from N, O, and S; The pharmaceutically acceptable salts include alkali metal salts, optionally sodium salts.

2. The compound according to claim 1, characterized in that, The Ar 1 and Ar 2 Each is independently selected from unsubstituted or R-substituted phenyl, naphthyl, biphenyl, benzothiazolyl, benzothiophene, benzofuranyl, indolyl, quinolinyl, isoquinolinyl, thiazolyl, thiadiazolyl, pyridyl, C3-C8 cycloalkyl or C3-C8 heterocyclic groups.

3. The compound according to claim 1 or 2, characterized in that, Each of the R groups is independently selected from hydroxyl, amino, fluorine, bromine, cyano, carboxyl, methoxy, methylthio, methyl, ethyl, propyl, ethoxycarbonyl, carbamoyl, sulfonic acid, sodium sulfonate, oxo, methylenedioxy, or -C(=O)NH2.

4. The compound according to any one of claims 1 to 3, characterized in that, The L 1 and L 2 Each is independently selected from single bonds, -CH2-, -CH=CH-, -O-, -S-, -NH-, -C(=O)-, -C(=S)-, -SO2-, -N=N-, -OCH2CH2O-, -OCH2CH2CH2O-, or divalent linkages formed by sequentially connecting two or three of these linkages.

5. The compound according to any one of claims 1 to 4, characterized in that, X is selected from non-existent, -O-, -NH-, -C(=S)-, -N=N-, 5-7 membered heterocyclic groups containing N and S, or fused heterocyclic groups containing N, S and oxo groups.

6. The compound according to any one of claims 1 to 5, characterized in that, The compound is selected from any of the following classes of compounds: Nitrogen- and / or sulfur-containing heterocyclic compounds; Fused aromatic ring compounds; Polyhydroxyaryl thioamide compounds; Aromatic compounds containing carboxyl aryl ethers and polyether linkages; or Azosulfonate compounds.

7. The compound according to any one of claims 1 to 6, characterized in that, The compound is selected from compounds of formula I, II, III, IV, V, VI or VII, or their tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts or prodrugs: 、 、 、 、 、 、 。 8. A pharmaceutical composition, characterized in that, Include: The compound of any one of claims 1 to 7, or its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug; The pharmaceutical composition is used for the prevention and / or treatment of diseases associated with abnormal oleyl-ACP hydrolase activity; Optionally, the disease associated with abnormal oleyl-ACP hydrolase activity is respiratory syncytial virus infection; Optionally, the pharmaceutical composition further includes a pharmaceutically acceptable carrier, excipient, or excipient.

9. The use of the compound of any one of claims 1 to 7, or its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, or the pharmaceutical composition of claim 8 in the preparation of an oleyl-ACP hydrolase inhibitor; Optionally, the use is for in vitro use for non-therapeutic purposes.

10. The use of any one of the compounds of claims 1 to 7, or tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts or prodrugs thereof, or the pharmaceutical composition of claim 8, in the preparation of a medicament for the prevention and / or treatment of diseases associated with abnormal oleyl-ACP hydrolase activity or expression levels; Optionally, the disease associated with abnormal oleyl-ACP hydrolase activity or expression levels is a viral infection; Optionally, the viral infection is respiratory syncytial virus infection.

11. The use of any compound of claims 1 to 7, or a tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug thereof, or the pharmaceutical composition of claim 8, in the preparation of a medicament or composition for inhibiting respiratory virus replication; Optionally, the respiratory virus is respiratory syncytial virus.

12. The use of any compound of claims 1 to 7, or a tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug thereof, in the preparation of a kit for detecting oleyl-ACP hydrolase activity or screening oleyl-ACP hydrolase inhibitors; Optionally, the kit includes the compound, oleyl-ACP hydrolase or an enzyme preparation containing oleyl-ACP hydrolase, a reaction buffer, and a detection substrate.

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