A p-methoxyphenylamino cyclobutenedione quinoline derivative, and a preparation method and application thereof

Abstract

The application discloses a p-methoxyphenylamino cyclobutenedione quinoline derivative, a preparation method and application thereof, and the derivative has a structure shown in formula I. 14 The derivative has good P2Y 14 R antagonistic activity, and obviously improves apparent solubility in water; application in treatment of liver fibrosis can obviously reduce liver inflammation, tissue structure damage and abnormal accumulation of collagen fibers, and has an oral effective characteristic, is favorable for improving patient compliance, and has a very good clinical application prospect.

Description

Technical Field

[0001] This invention relates to a p-methoxyphenylaminocyclobutenidine quinoline derivative, and also to a method for preparing the above derivative and its application in the preparation of P2Y. 14 Applications of R antagonists. Background Technology

[0002] Liver fibrosis, a key pathological process in the progression of chronic liver disease to cirrhosis and even liver cancer, has long lacked safe and effective methods for reversing. Currently, clinical anti-fibrotic therapy faces two major challenges: first, existing drugs lack sufficient targeting of the disease's core driver cells—hepatic stellate cells (HSCs), resulting in limited efficacy and significant side effects; second, apart from liver transplantation, there is no therapy that can truly reverse the fibrotic process. Therefore, identifying key targets that can precisely regulate HSC function and developing corresponding therapeutic drugs has become a pressing scientific challenge in the field of liver disease research.

[0003] P2Y 14 As a unique member of the purinergic receptor family, the P2Y receptor has been identified in recent years as a key molecule regulating the progression of liver fibrosis. Unlike classic purinergic receptors, P2Y receptors... 14 The receptor specifically recognizes glyconucleotide ligands such as uridine diphosphate glucose (UDP-Glc), and this unique ligand selectivity enables it to play a special role in metabolic stress and cell communication. In the normal liver, P2Y 14 The receptor expression level is low. However, when the liver is subjected to viral invasion, alcohol damage, lipid accumulation, or autoimmune attack, this receptor exhibits specific high expression on the surface of HSCs. More importantly, UDP-Glc released by damaged hepatocytes acts as a "danger signal," interacting with P2Y on HSCs. 14 Upon receptor binding, a cascade reaction is triggered: through the Gi / o protein signaling pathway, downstream key signaling molecules such as MAPK and ERK are activated, causing quiescent HSCs to transform into highly activated myofibroblasts. These activated cells not only synthesize large amounts of extracellular matrix components such as type I and type III collagen, but also form a vicious cycle by secreting pro-fibrotic factors, ultimately leading to liver structural damage and loss of function.

[0004] Based on P2Y 14 The receptor is specifically and highly expressed on activated hepatocytes (HSCs), thus providing a precise molecular target for the development of anti-fibrotic drugs. Its inhibitors can selectively act on fibrotic core cells, effectively avoiding damage to normal hepatocytes and significantly improving treatment safety. As an upstream hub connecting hepatocyte damage and HSC activation, targeting P2Y... 14 Receptors can block fibrosis signaling at its source, offering greater potential for fundamental treatment compared to interventions targeting downstream collagen deposition. Preclinical studies have further confirmed that inhibiting P2Y...14 The receptor can not only prevent the formation of new fibrous tissue, but also induce apoptosis of activated HSCs and promote their phenotypic reversal, thereby creating favorable conditions for the degradation of deposited fibrous tissue and demonstrating the comprehensive advantages of multiple therapeutic benefits.

[0005] There is currently no P2Y globally. 14 Most R-related drugs that are marketed or in clinical trials are in the preclinical or drug discovery stage, mainly including naphthoic acid derivatives, amide derivatives, triazole derivatives, and benzo[a]heterocyclic derivatives. These compounds are mostly derived from the only currently marketed P2Y. 14 The R inhibitor PPTN, after structural optimization, possesses anti-inflammatory and immunomodulatory activities, but its physicochemical properties are poor, rendering it unsuitable for drug development. Furthermore, recently reported quinoline squaring amide derivatives have achieved drug-like effects on P2Y through structural optimization. 14 R exhibits nanomolar activity and high selectivity, demonstrating good efficacy in various inflammation models. However, the development of these compounds faces multiple challenges, including poor water solubility and the need for improvement in pharmacokinetic properties. Therefore, further optimization based on their structure is necessary to obtain novel P2Y compounds with good water solubility and high bioavailability. 14 R inhibitors are of great significance for targeted treatment of liver fibrosis. Summary of the Invention

[0006] Objective of the Invention: The objective of this invention is to provide a p-methoxyphenylaminocyclobutenidine quinoline derivative with good water solubility and high bioavailability, and also to provide a method for preparing the above derivative and its application in the preparation of P2Y. 14 Applications of R antagonists.

[0007] Technical solution: The p-methoxyphenylaminocyclobutenidine quinoline derivative of the present invention has the structure shown in Formula I: ;

[0008] R¹ or R² is selected from alkyl groups (e.g., methyl, ethyl, n-propyl) or 5- to 6-membered heterocyclic groups containing an alkyl chain with a length of 2 to 3 carbon atoms.

[0009] Wherein, R1 or R2 is an unsubstituted methyl, ethyl, propyl, or alkylene chain (-(CH2)). n The linker is a pentacyclic ring (tetrahydropyrrole) or a six-membered ring (piperidine, morpholine, tetrahydropyran), ethyl carboxylic acid, propanol, methyl carboxylic acid, ethanol, or 2-methoxyethoxy; wherein the pentacyclic ring is tetrahydropyrrole and the six-membered ring is piperidine, morpholine, or tetrahydropyran.

[0010] The above derivatives include the following structures: ; .

[0011] The above method for preparing the p-methoxyphenylaminocyclobutenedionylquinoline derivative involves reacting a compound of formula II with a compound of formula III to obtain the p-methoxyphenylaminocyclobutenedionylquinoline derivative shown in formula I: ;

[0012] Its synthetic route is as follows: .

[0013] The present invention also discloses a pharmaceutical composition comprising the above-mentioned derivative or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient.

[0014] The above-mentioned derivatives or pharmaceutical compositions can be used in the preparation of P2Y. 14 In drugs used to treat receptor-related diseases.

[0015] Wherein, the P2Y 14 Receptor-related diseases include ischemic acute kidney injury, nerve damage following subarachnoid hemorrhage, inflammatory diseases such as gout, venous thromboembolism, and liver fibrosis.

[0016] The aforementioned derivatives or pharmaceutical compositions can be used in the preparation of drugs for treating liver fibrosis and can be used in combination with other drugs for treating related diseases. The derivatives or pharmaceutical compositions described in this invention can be combined with anti-inflammatory and hepatoprotective drugs (such as silymarin, bicyclol, glycyrrhizic acid preparations, ursodeoxycholic acid, etc.) to reduce inflammatory damage at the source and block the initiation of fibrosis; they can be combined with drugs for treating metabolic-related fatty liver disease (such as PPAR agonists, FXR agonists) to simultaneously intervene in metabolic disorders and fibrotic deposition; they can be combined with targeted anti-fibrotic drugs (such as LOXL2, integrins, Galectin-3 inhibitors) to block hepatic stellate cell activation and extracellular matrix deposition from multiple angles; and they can be combined with renin-angiotensin system blockers (such as losartan) to dual inhibit the pro-fibrotic effect of angiotensin II.

[0017] Invention Principle: The p-methoxyphenylaminocyclobutenedionylquinoline derivative of this invention utilizes the alkylation of the amino group in the square amide skeleton to block the formation of strong hydrogen bonds in the compound molecule, thus synthesizing the p-methoxyphenylaminocyclobutenedionylquinoline derivative of this invention. This overcomes the problem in the prior art where the formation of strong hydrogen bonds between the secondary square amides in the square amide skeleton leads to high melting points and poor solubility in the compounds. Compared with the previously studied compound HDB-18, its solubility is significantly improved. Formula 14, where R1 is N-(propyl)morpholino and R2 is a hydrogen atom, maintains the inhibitory activity of HDB-18 while increasing solubility by 100 times, and can then be applied to P2Y.14 In drugs for treating receptor-related diseases, especially those for liver fibrosis, stronger solubility can be achieved, resulting in a stronger therapeutic effect.

[0018] Beneficial effects: Compared with the prior art, the present invention has the following significant advantages: the p-methoxyphenylaminocyclobutenedionylquinoline derivative of the present invention has better P2Y. 14 The compound exhibits antagonistic activity against R and significantly improves solubility in water. The best-performing compound showed an inhibitory activity of 119.87% and a solubility of 675.15 μg / ml. In the treatment of liver fibrosis, it can significantly reduce liver inflammation, tissue damage, and abnormal collagen fiber accumulation. Its oral efficacy improves patient compliance and makes it highly promising for clinical application. Furthermore, it shows significant efficacy in a mouse model of liver fibrosis, downregulating key fibrosis genes (such as ACTA2 and COL1A1), reducing ALT / AST enzyme activity, and alleviating liver damage. It also reduces liver TG / TC levels and improves lipid metabolism disorders. Therefore, this invention has oral efficacy, which improves patient compliance and makes it highly promising for clinical application. Attached Figure Description

[0019] Figure 1 This study investigated the effect of compound LJR-8 on serum ALT and AST levels in a CDF-HFD-induced liver fibrosis mouse model. All data are expressed as mean ± standard error (SEM). *Compared with the CDF-HFD group* P < 0.05, ** P < 0.01, #Compared with the control group, # P < 0.05, ## P < 0.01, ### P < 0.001.

[0020] Figure 2 This study investigated the effect of compound LJR-8 on serum TG and TC levels in a CDF-HFD-induced liver fibrosis mouse model. All data are expressed as mean ± standard error (SEM). *Compared with the CDF-HFD group* P < 0.05, ** P < 0.01, #Compared with the control group, # P < 0.05, ## P < 0.01, ### P < 0.001.

[0021] Figure 3 This study investigated the effect of compound LJR-8 on hydroxyproline levels in a CDF-HFD-induced liver fibrosis mouse model. All data are expressed as mean ± standard error (SEM). *Compared with the CDA-HFD group* P < 0.05, ** P < 0.01, #Compared with the control group, # P < 0.05, ## P < 0.01, ### P < 0.001.

[0022] Figure 4This study investigated the effect of compound LJR-8 on the mRNA expression levels of fibrosis-related marker genes in a CDF-HFD-induced liver fibrosis mouse model. All data are expressed as mean ± standard error (SEM). *Compared with the CDF-HFD group* P < 0.05, ** P < 0.01, #Compared with the control group, # P < 0.05, ## P < 0.01, ### P < 0.001.

[0023] Figure 5 Masson's staining bar chart shows the liver effects of compound LJR-8 on CDF-HFD-induced diet-induced mice. All data are expressed as mean ± standard error (SEM). *Compared with CDA-HFD group* P < 0.05, ** P < 0.01, #Compared with control group, # P < 0.05, ## P < 0.01, ### P < 0.001.

[0024] Figure 6 HE staining of liver tissue from CDF-HFD-induced diet-induced mice treated with compound LJR-8. All data are expressed as mean ± standard error (SEM). *Compared with CDA-HFD group* P < 0.05, ** P < 0.01, #Compared with control group, # P < 0.05, ## P < 0.01, ### P < 0.001. Detailed Implementation

[0025] The technical solution of the present invention will be further described below with reference to the embodiments. The test materials used in the embodiments can all be purchased through conventional means.

[0026] Example 1

[0027] Synthesis of 3-((4-methoxyphenyl)(methyl)amino)-4-(quinolin-6-ylamino)cyclobut-3-ene-1,2-dione:

[0028] 1.34 g of quinoline-6-amine was dissolved in methanol, and 1.42 g of 3,4-dimethoxycyclobutane-3-ene-1,2-dione was added. The mixture was stirred at room temperature for 24 h. When the reaction was complete, the crude product was extracted with water and ethyl acetate. The organic phase was dried over anhydrous sodium sulfate and evaporated to dryness under reduced pressure. The product was purified by silica gel column chromatography, eluting with dichloromethane:methanol (50:1) to give 3-(quinoline-6-ylamino)-4-methoxycyclobutane-3-ene-1,2-dione.

[0029] 0.738 g of p-methoxyaniline was dissolved in acetonitrile, and 0.846 g of iodomethane and 0.696 g of potassium iodide were added. The mixture was stirred at 65 °C for 30 minutes, followed by the addition of 0.828 g of potassium carbonate. The mixture was then refluxed at 70 °C and stirred for 12 hours. When the reaction was complete, the crude product was extracted with water and ethyl acetate. The organic phase was dried over anhydrous sodium sulfate and evaporated to dryness under reduced pressure. The product was purified by silica gel column chromatography, eluting with dichloromethane:methanol (20:1) to give 4-methoxy-N-methylaniline.

[0030] 0.642 g of 3-(quinoline-6-ylamino)-4-methoxycyclobut-3-ene-1,2-dione was dissolved in methanol, and 0.642 g of 4-methoxy-N-methylaniline was added. The mixture was stirred at 65 °C for 24 hours. When the reaction was complete, the crude product was extracted with water and ethyl acetate. The organic phase was dried over anhydrous sodium sulfate and evaporated to dryness under reduced pressure. The product was purified by silica gel column chromatography, eluting with dichloromethane:methanol (10:1) to give 3-((4-methoxyphenyl)(methyl)amino)-4-(quinoline-6-ylamino)cyclobut-3-ene-1,2-dione.

[0031] The NMR data are as follows:

[0032] 1H NMR (600 MHz, DMSO-d6) δ 8.85 (dd, J = 7.5, 1.6 Hz, 1H), 8.54 (dd,J = 7.5, 1.6 Hz, 1H), 8.26 (dt, J = 7.5, 1.5 Hz, 1H), 7.89 (d, J = 7.5 Hz,1H), 7.38 (t, J = 7.5 Hz, 1H), 6.92 (d, J = 1.6 Hz, 1H), 6.76 (s, 4H), 3.79(s, 3H), 3.47 (s, 3H).

[0033] Example 2

[0034] Synthesis of 3-(ethyl(4-methoxyphenyl)amino)-4-(quinolin-6-ylamino)cyclobut-3-ene-1,2-dione:

[0035] The synthesis method using iodoethane is described in Example 1.

[0036] The NMR data are as follows:

[0037] 1H NMR (600 MHz, DMSO-d6) δ 8.85 (dd, J = 7.5, 1.5 Hz, 1H), 8.53 (dd,J = 7.6, 1.4 Hz, 1H), 8.26 (dt, J = 7.4, 1.6 Hz, 1H), 7.89 (d, J = 7.5 Hz,1H), 7.38 (t, J = 7.5 Hz, 1H), 6.93 (t, J = 1.6 Hz, 1H), 6.79 – 6.73 (m, 2H), 6.73 – 6.67 (m, 2H), 4.56 (q, J = 8.0 Hz, 2H), 3.79 (s, 3H), 1.50 (t, J = 8.0Hz, 3H).

[0038] Example 3

[0039] Synthesis of 3-((4-methoxyphenyl)(propyl)amino)-4-(quinolin-6-ylamino)cyclobut-3-ene-1,2-dione:

[0040] The synthesis method using 1-bromopropane as a raw material is described in Example 1.

[0041] The NMR data are as follows:

[0042] 1H NMR (600 MHz, DMSO-d6) δ 8.85 (dd, J = 7.5, 1.5 Hz, 1H), 8.39 (dd,J = 7.6, 1.4 Hz, 1H), 8.27 (dt, J = 7.6, 1.5 Hz, 1H), 7.88 (d, J = 7.5 Hz,1H), 7.38 (t, J = 7.5 Hz, 1H), 6.95 (t, J = 1.4 Hz, 1H), 6.79 – 6.73 (m, 2H), 6.72 – 6.66 (m, 2H), 3.81 – 3.74 (m, 5H), 1.62 (h, J = 7.9 Hz, 2H), 0.93 (t,J = 8.0 Hz, 3H).

[0043] Example 4

[0044] Synthesis of 3-((2-hydroxyethyl)(4-methoxyphenyl)amino)-4-(quinoline-6-ylamino)cyclobut-3-ene-1,2-dione:

[0045] The synthesis method using 2-bromoethanol as a raw material is described in Example 1.

[0046] The NMR data are as follows:

[0047] 1H NMR (600 MHz, DMSO-d6) δ 8.85 (dd, J = 7.5, 1.5 Hz, 1H), 8.50 (dd,J = 7.5, 1.6 Hz, 1H), 8.27 (dt, J = 7.7, 1.6 Hz, 1H), 7.89 (d, J = 7.5 Hz,1H), 7.38 (t, J = 7.5 Hz, 1H), 6.98 (t, J = 1.4 Hz, 1H), 6.79 – 6.73 (m, 2H), 6.71 – 6.66 (m, 2H), 4.88 (t, J = 4.9 Hz, 1H), 3.79 (s, 3H), 3.57 (dt, J =5.1, 3.8 Hz, 2H), 3.29 (t, J = 3.8 Hz, 2H).

[0048] Example 5

[0049] Synthesis of N-(3,4-diketone-2-(quinolin-6-ylamino)cyclobut-1-en-1-yl)-N-(4-methoxyphenyl)glycine:

[0050] The synthesis method using 2-bromoacetic acid as a raw material is described in Example 1.

[0051] The NMR data are as follows:

[0052] 1H NMR (600 MHz, DMSO-d6) δ 8.85 (dd, J = 7.5, 1.5 Hz, 1H), 8.40 (dd,J = 7.5, 1.6 Hz, 1H), 8.31 (dt, J = 7.5, 1.6 Hz, 1H), 7.88 (d, J = 7.5 Hz,1H), 7.38 (t, J = 7.5 Hz, 1H), 7.22 (t, J = 1.5 Hz, 1H), 6.80 – 6.74 (m, 4H), 4.58 (s, 2H), 3.79 (s, 3H).

[0053] Example 6

[0054] Synthesis of 3-((4-methoxyphenyl)(2-(pyrrolidone-1-yl)ethyl)amino)-4-(quinolin-6-ylamino)cyclobut-3-ene-1,2-dione:

[0055] The synthesis method using 1-(2-bromoethyl)pyrrolidine as a raw material is described in Example 1.

[0056] The NMR data are as follows:

[0057] 1H NMR (600 MHz, DMSO-d6) δ 8.85 (dd, J = 7.5, 1.6 Hz, 1H), 8.34 (dt,J = 7.5, 1.6 Hz, 1H), 8.02 (dd, J = 7.5, 1.3 Hz, 1H), 7.85 (d, J = 7.5 Hz,1H), 7.54 (d, J = 1.6 Hz, 1H), 7.38 (t, J = 7.6 Hz, 1H), 6.81 – 6.74 (m, 4H), 3.79 (s, 3H), 3.54 (t, J = 5.9 Hz, 2H), 3.29 – 3.21 (m, 4H), 2.50 (t, J = 6.0Hz, 2H), 1.95 – 1.87 (m, 4H).

[0058] Example 7

[0059] Synthesis of 3-((4-methoxyphenyl)(2-morpholinoethyl)amino)-4-(quinolin-6-ylamino)cyclobut-3-ene-1,2-dione:

[0060] The synthesis method using 4-(2-bromoethyl)morpholine as a raw material is described in Example 1.

[0061] The NMR data are as follows:

[0062] 1H NMR (600 MHz, DMSO-d6) δ 8.85 (dd, J = 7.5, 1.5 Hz, 1H), 8.52 (dd,J = 7.6, 1.4 Hz, 1H), 8.25 (dt, J = 7.5, 1.6 Hz, 1H), 7.90 (d, J = 7.5 Hz,1H), 7.38 (t, J = 7.5 Hz, 1H), 6.94 (t, J = 1.7 Hz, 1H), 6.79 – 6.73 (m, 2H), 6.71 – 6.66 (m, 2H), 3.79 (s, 3H), 3.57 – 3.51 (m, 3H), 3.54 (s, 3H), 2.50(t, J = 6.1 Hz, 2H), 2.41 (t, J = 4.6 Hz, 4H).

[0063] Example 8

[0064] Synthesis of 3-((4-methoxyphenyl)(2-(tetrahydro-2H-pyran-4-yl)ethyl)amino)-4-(quinoline-6-ylamino)cyclobut-3-ene-1,2-dione:

[0065] The synthesis method is described in Example 1, using 4-(2-bromoethyl)tetrahydro-2H-pyran as the starting material.

[0066] The NMR data are as follows:

[0067] 1H NMR (600 MHz, DMSO-d6) δ 8.85 (dd, J = 7.5, 1.5 Hz, 1H), 8.46 (dd,J = 7.5, 1.5 Hz, 1H), 8.27 (dt, J = 7.5, 1.5 Hz, 1H), 7.89 (d, J = 7.5 Hz,1H), 7.38 (t, J = 7.5 Hz, 1H), 6.92 (d, J = 1.6 Hz, 1H), 6.79 – 6.73 (m, 2H),6.72 – 6.66 (m, 2H), 3.88 (dt, J = 11.4, 7.1 Hz, 2H), 3.81 – 3.74 (m, 5H), 3.45 (dt, J = 11.5, 7.1 Hz, 2H), 1.61 – 1.52 (m, 4H), 1.40 (hept, J = 6.9 Hz, 1H), 1.22 (dq, J = 13.9, 7.1 Hz, 2H).

[0068] Example 9

[0069] Synthesis of 3-((4-methoxyphenyl)(2-(piperidin-1-yl)ethyl)amino)-4-(quinolin-6-ylamino)cyclobut-3-ene-1,2-dione:

[0070] The synthesis method using 1-(2-bromoethyl)piperidine as a raw material is described in Example 1.

[0071] The NMR data are as follows:

[0072] 1H NMR (600 MHz, DMSO-d6) δ 8.85 (dd, J = 7.5, 1.5 Hz, 1H), 8.53 (dd,J = 7.5, 1.5 Hz, 1H), 8.26 (dt, J = 7.5, 1.6 Hz, 1H), 7.89 (d, J = 7.4 Hz,1H), 7.38 (t, J = 7.5 Hz, 1H), 7.00 (d, J = 1.4 Hz, 1H), 6.79 – 6.73 (m, 2H), 6.71 – 6.66 (m, 2H), 3.79 (s, 3H), 3.54 (t, J = 6.3 Hz, 2H), 2.50 (t, J = 6.4Hz, 2H), 2.42 (t, J = 5.4 Hz, 4H), 1.52 – 1.40 (m, 5H), 1.40 (d, J = 5.8 Hz, 1H).

[0073] Example 10

[0074] Synthesis of 3-((3-hydroxypropyl)(4-methoxyphenyl)amino)-4-(quinolin-6-ylamino)cyclobut-3-ene-1,2-dione:

[0075] The synthesis method using 3-bromoprop-1-ol as a starting material is described in Example 1.

[0076] The NMR data are as follows:

[0077] 1H NMR (600 MHz, DMSO-d6) δ 8.85 (dd, J = 7.5, 1.5 Hz, 1H), 8.34 (dt,J = 7.5, 1.6 Hz, 1H), 8.02 (dd, J = 7.5, 1.5 Hz, 1H), 7.86 (d, J = 7.5 Hz,1H), 7.49 (t, J = 1.7 Hz, 1H), 7.38 (t, J = 7.6 Hz, 1H), 6.79 – 6.73 (m, 2H), 6.72 – 6.67 (m, 2H), 4.55 (t, J = 5.1 Hz, 1H), 3.81 – 3.74 (m, 5H), 3.53 (q,J = 4.8 Hz, 2H), 1.75 (tt, J = 7.4, 4.6 Hz, 2H).

[0078] Example 11

[0079] Synthesis of 3-((3-hydroxybut-3-en-1-yl)(4-methoxyphenyl)amino)-4-(quinoline-6-ylamino)cyclobut-3-en-1,2-dione:

[0080] The synthesis method is described in Example 1, using 4-bromobut-1-en-2-ol as the starting material.

[0081] The NMR data are as follows:

[0082] 1H NMR (600 MHz, DMSO-d6) δ 8.85 (dd, J = 7.5, 1.5 Hz, 1H), 8.48 (dd,J = 7.5, 1.5 Hz, 1H), 8.21 (dt, J = 7.5, 1.5 Hz, 1H), 7.88 (d, J = 7.5 Hz,1H), 7.38 (t, J = 7.5 Hz, 1H), 6.96 – 6.91 (m, 2H), 6.82 – 6.73 (m, 3H), 3.79(s, 3H), 3.46 (t, J = 7.9 Hz, 2H), 2.51 (t, J = 7.9 Hz, 2H).

[0083] Example 12

[0084] Synthesis of 3-((4-methoxyphenyl)(3-(pyrrolidone-1-yl)propyl)amino)-4-(quinolin-6-ylamino)cyclobut-3-ene-1,2-dione:

[0085] The synthesis method using 1-(3-bromopropyl)pyrrolidine as a starting material is described in Example 1.

[0086] The NMR data are as follows:

[0087] 1H NMR (600 MHz, DMSO-d6) δ 8.85 (dd, J = 7.5, 1.5 Hz, 1H), 8.34 (dt,J = 7.5, 1.6 Hz, 1H), 7.84 – 7.78 (m, 2H), 7.52 (d, J = 1.4 Hz, 1H), 7.38 (t,J = 7.5 Hz, 1H), 6.92 – 6.87 (m, 2H), 6.79 – 6.73 (m, 2H), 3.81 – 3.74 (m,5H), 2.70 – 2.62 (m, 4H), 2.36 (t, J = 5.2 Hz, 2H), 2.00 – 1.91 (m, 6H).

[0088] Example 13

[0089] Synthesis of 3-((4-methoxyphenyl)(3-(piperidin-1-yl)propyl)amino)-4-(quinolin-6-ylamino)cyclobut-3-ene-1,2-dione:

[0090] The synthesis method using 1-(3-bromopropyl)piperidine as a raw material is described in Example 1.

[0091] The NMR data are as follows:

[0092] 1H NMR (600 MHz, DMSO-d6) δ 8.85 (dd, J = 7.5, 1.5 Hz, 1H), 8.43 (dd,J = 7.6, 1.4 Hz, 1H), 8.25 (dt, J = 7.5, 1.5 Hz, 1H), 7.38 (t, J = 7.5 Hz,1H), 6.97 (t, J = 1.5 Hz, 1H), 6.78 – 6.74 (m, 2H), 6.70 – 6.65 (m, 2H), 3.81– 3.74 (m, 5H), 2.42 (t, J = 5.4 Hz, 4H), 2.36 (t, J = 7.4 Hz, 2H), 1.95 (q,J = 7.5 Hz, 2H), 1.52 (p, J = 5.6 Hz, 4H), 1.41 (q, J = 6.1 Hz, 2H).

[0093] Example 14

[0094] Synthesis of 3-((4-methoxyphenyl)(3-morpholinopropyl)amino)-4-(quinolin-6-ylamino)cyclobut-3-ene-1,2-dione:

[0095] The synthesis method using 1-(3-bromopropyl)piperidine as a raw material is described in Example 1.

[0096] The NMR data are as follows:

[0097] 1H NMR (600 MHz, DMSO-d6) δ 8.85 (dd, J = 7.5, 1.5 Hz, 1H), 8.46 (dd,J = 7.5, 1.5 Hz, 1H), 8.25 (dt, J = 7.5, 1.6 Hz, 1H), 7.89 (d, J = 7.5 Hz,1H), 7.38 (t, J = 7.5 Hz, 1H), 6.94 (t, J = 1.5 Hz, 1H), 6.79 – 6.73 (m, 2H), 6.72 – 6.67 (m, 2H), 3.81 – 3.74 (m, 5H), 3.56 (t, J = 4.7 Hz, 4H), 2.39 –2.32 (m, 6H), 1.94 (p, J = 7.5 Hz, 2H).

[0098] Example 15

[0099] Synthesis of 3-((2-(2-methoxyethoxy)ethyl)(4-methoxyphenyl)amino)-4-(quinoline-6-ylamino)cyclobut-3-ene-1,2-dione:

[0100] The synthesis method is described in Example 1, using 1-bromo-2-(2-methoxyethoxy)ethane as the starting material.

[0101] The NMR data are as follows:

[0102] 1H NMR (600 MHz, DMSO-d6) δ 8.85 (dd, J = 7.5, 1.5 Hz, 1H), 8.43 (dd,J = 7.5, 1.6 Hz, 1H), 8.28 (dt, J = 7.5, 1.6 Hz, 1H), 7.88 (d, J = 7.5 Hz,1H), 7.38 (t, J = 7.5 Hz, 1H), 6.79 – 6.68 (m, 4H), 3.79 (s, 3H), 3.66 – 3.58(m, 4H), 3.54 (td, J = 7.0, 1.2 Hz, 2H), 3.31 – 3.25 (m, 5H).

[0103] Example 16

[0104] Synthesis of 3-((4-methoxyphenyl)amino)-4-(methyl(quinolin-6-yl)amino)cyclobut-3-ene-1,2-dione:

[0105] 1.34 g of quinoline-6-amine was dissolved in acetonitrile, and 1.846 g of iodomethane and 1.696 g of potassium iodide were added. The mixture was stirred at room temperature for 30 minutes, followed by the addition of 1.828 g of potassium carbonate. The mixture was then refluxed at 70 °C and stirred for 12 hours. When the reaction was complete, the crude product was extracted with water and ethyl acetate. The organic phase was dried over anhydrous sodium sulfate and evaporated to dryness under reduced pressure. The product was purified by silica gel column chromatography, eluting with dichloromethane:methanol (50:1) to give N-methylquinoline-6-amine.

[0106] 0.642 g of N-methylquinoline-6-amine was dissolved in methanol, and 0.642 g of 3,4-dimethoxycyclobutane-3-ene-1,2-dione was added. The mixture was stirred at 65 °C for 24 h. When the reaction was complete, the crude product was extracted with water and ethyl acetate. The organic phase was dried over anhydrous sodium sulfate and evaporated to dryness under reduced pressure. The product was purified by silica gel column chromatography, eluting with dichloromethane:methanol (20:1) to give 3-methoxy-4-(methyl(quinoline-6-yl)amino)cyclobutane-3-ene-1,2-dione.

[0107] 0.642 g of p-toluidine was dissolved in methanol, and 0.642 g of 3-methoxy-4-(methyl(quinoline-6-yl)amino)cyclobut-3-ene-1,2-dione was added. The mixture was stirred at 65 °C for 24 hours. When the reaction was complete, the crude product was extracted with water and ethyl acetate. The organic phase was dried over anhydrous sodium sulfate and evaporated to dryness under reduced pressure. The product was purified by silica gel column chromatography, eluting with dichloromethane:methanol (10:1) to give 3-((4-methoxyphenyl)amino)-4-(methyl(quinoline-6-yl)amino)cyclobut-3-ene-1,2-dione.

[0108] The NMR data are as follows:

[0109] 1H NMR (600 MHz, DMSO-d6) δ 8.85 (dd, J = 7.5, 1.5 Hz, 1H), 8.29 (dt,J = 7.6, 1.5 Hz, 1H), 8.08 (dd, J = 7.5, 1.6 Hz, 1H), 7.88 (d, J = 7.5 Hz,1H), 7.38 (t, J = 7.5 Hz, 1H), 7.20 (s, 1H), 6.81 – 6.75 (m, 2H), 6.69 – 6.64(m, 2H), 3.79 (s, 3H), 3.47 (s, 3H).

[0110] Example 17

[0111] Synthesis of 3-(ethyl(quinoline-6-yl)amino)-4-((4-methoxyphenyl)amino)cyclobut-3-ene-1,2-dione:

[0112] The synthesis method using iodoethane is described in Example 17.

[0113] The NMR data are as follows:

[0114] 1H NMR (600 MHz, DMSO-d6) δ 8.85 (dd, J = 7.5, 1.5 Hz, 1H), 8.29 (dt,J = 7.6, 1.5 Hz, 1H), 7.98 (dd, J = 7.6, 1.4 Hz, 1H), 7.86 (d, J = 7.5 Hz,1H), 7.38 (t, J = 7.5 Hz, 1H), 7.18 (t, J = 1.5 Hz, 1H), 6.77 – 6.71 (m, 2H), 6.69 – 6.63 (m, 2H), 4.56 (q, J = 8.0 Hz, 2H), 3.79 (s, 3H), 1.50 (t, J = 8.0Hz, 3H).

[0115] Example 18

[0116] Synthesis of 3-((4-methoxyphenyl)amino)-4-(propyl(quinolin-6-yl)amino)cyclobut-3-ene-1,2-dione:

[0117] The synthesis method using 1-bromopropane as a raw material is described in Example 17.

[0118] The NMR data are as follows:

[0119] 1H NMR (600 MHz, DMSO-d6) δ 8.85 (dd, J = 7.5, 1.5 Hz, 1H), 8.28 (dt,J = 7.5, 1.6 Hz, 1H), 7.95 (dd, J = 7.6, 1.4 Hz, 1H), 7.85 (d, J = 7.7 Hz,1H), 7.38 (t, J = 7.5 Hz, 1H), 7.15 (t, J = 1.6 Hz, 1H), 6.73 – 6.68 (m, 2H), 6.69 – 6.63 (m, 2H), 3.81 – 3.74 (m, 5H), 1.62 (h, J = 7.9 Hz, 2H), 0.93 (t,J = 8.0 Hz, 3H).

[0120] Example 19

[0121] Synthesis of 3-((4-methoxyphenyl)amino)-4-(propyl(quinolin-6-yl)amino)cyclobut-3-ene-1,2-dione:

[0122] The synthesis method using 2-bromoethanol as a raw material is described in Example 17.

[0123] The NMR data are as follows:

[0124] 1H NMR (600 MHz, DMSO-d6) δ 8.85 (dd, J = 7.5, 1.5 Hz, 1H), 8.26 (dt,J = 7.5, 1.5 Hz, 1H), 7.99 (dd, J = 7.5, 1.6 Hz, 1H), 7.86 (d, J = 7.7 Hz,1H), 7.38 (t, J = 7.5 Hz, 1H), 6.80 – 6.74 (m, 2H), 6.69 – 6.63 (m, 2H), 4.84(t, J = 5.1 Hz, 1H), 3.79 (s, 3H), 3.57 (dt, J = 5.1, 3.8 Hz, 2H), 3.29 (t, J = 3.8 Hz, 2H).

[0125] Example 21

[0126] Synthesis of 3-((2-hydroxypropenyl)(quinoline-6-yl)amino)-4-((4-methoxyphenyl)amino)cyclobut-3-ene-1,2-dione:

[0127] The synthesis method using 2-bromoacetic acid as a raw material is described in Example 17.

[0128] The NMR data are as follows:

[0129] 1H NMR (600 MHz, DMSO-d6) δ 8.85 (dd, J = 7.5, 1.5 Hz, 1H), 8.32 (dt,J = 7.6, 1.5 Hz, 1H), 7.93 (dd, J = 7.5, 1.6 Hz, 1H), 7.86 (d, J = 7.5 Hz,1H), 7.38 (t, J = 7.5 Hz, 1H), 7.32 (t, J = 1.4 Hz, 1H), 6.89 – 6.83 (m, 2H), 6.69 – 6.63 (m, 2H), 4.58 (s, 2H), 3.79 (s, 3H).

[0130] Example 20

[0131] Synthesis of 3-((2-cyclopentylethyl)(quinolin-6-yl)amino)-4-((4-methoxyphenyl)amino)cyclobut-3-ene-1,2-dione:

[0132] The synthesis method using 1-(2-bromoethyl)pyrrolidine as a raw material is described in Example 17.

[0133] The NMR data are as follows:

[0134] 1H NMR (600 MHz, DMSO-d6) δ 8.85 (dd, J = 7.5, 1.5 Hz, 1H), 8.30 (dt,J = 7.8, 1.6 Hz, 1H), 8.00 (dd, J = 7.5, 1.5 Hz, 1H), 7.86 (d, J = 7.4 Hz,1H), 7.38 (t, J = 7.5 Hz, 1H), 7.20 (t, J = 1.6 Hz, 1H), 6.69 – 6.59 (m, 4H), 3.79 (s, 3H), 3.54 (t, J = 6.1 Hz, 2H), 3.29 – 3.21 (m, 4H), 2.50 (t, J = 6.1Hz, 2H), 1.95 – 1.87 (m, 4H).

[0135] Example 21

[0136] Synthesis of 3-((4-methoxyphenyl)amino)-4-((2-(piperidin-1-yl)ethyl)(quinolin-6-yl)amino)cyclobut-3-ene-1,2-dione:

[0137] The synthesis method using 1-(2-bromoethyl)piperidine as a raw material is described in Example 17.

[0138] The NMR data are as follows:

[0139] 1H NMR (600 MHz, DMSO-d6) δ 8.85 (dd, J = 7.5, 1.5 Hz, 1H), 8.29 (dt,J = 7.5, 1.5 Hz, 1H), 7.99 (dd, J = 7.5, 1.5 Hz, 1H), 7.86 (d, J = 7.5 Hz,1H), 7.38 (t, J = 7.5 Hz, 1H), 7.20 (t, J = 1.4 Hz, 1H), 6.69 – 6.59 (m, 4H),3.79 (s, 3H), 3.54 (t, J = 6.4 Hz, 2H), 2.50 (t, J = 6.4 Hz, 2H), 2.42 (t, J= 5.4 Hz, 4H), 1.48 (p, J = 5.6 Hz, 4H), 1.40 (d, J = 5.8 Hz, 1H).

[0140] Example 22

[0141] Synthesis of 3-((4-methoxyphenyl)amino)-4-((2-(piperidin-1-yl)ethyl)(quinolin-6-yl)amino)cyclobut-3-ene-1,2-dione:

[0142] The synthesis method using 4-(2-bromoethyl)morpholine as a raw material is described in Example 17.

[0143] The NMR data are as follows:

[0144] 1H NMR (600 MHz, DMSO-d6) δ 8.85 (dd, J = 7.5, 1.5 Hz, 1H), 8.30 (dt,J = 7.8, 1.5 Hz, 1H), 8.00 (dd, J = 7.5, 1.5 Hz, 1H), 7.87 (d, J = 7.4 Hz,1H), 7.38 (t, J = 7.5 Hz, 1H), 6.69 – 6.63 (m, 2H), 6.60 – 6.55 (m, 2H), 3.79(s, 3H), 3.57 – 3.51 (m, 6H), 2.50 (t, J = 6.3 Hz, 2H), 2.41 (t, J = 4.6 Hz, 4H).

[0145] Example 23

[0146] Synthesis of 3-((4-methoxyphenyl)amino)-4-((3-(piperrol-1-yl)propyl)(quinolin-6-yl)amino)cyclobut-3-ene-1,2-dione:

[0147] The synthesis method using 1-(3-bromopropyl)pyrrolidine as a starting material is described in Example 17.

[0148] The NMR data are as follows:

[0149] 1H NMR (600 MHz, DMSO-d6) δ 8.85 (dd, J = 7.5, 1.5 Hz, 1H), 8.28 (dt,J = 7.5, 1.6 Hz, 1H), 7.98 (dd, J = 7.5, 1.5 Hz, 1H), 7.86 (d, J = 7.4 Hz,1H), 7.38 (t, J = 7.5 Hz, 1H), 7.16 (t, J = 1.4 Hz, 1H), 6.69 – 6.63 (m, 2H), 6.59 – 6.54 (m, 2H), 3.81 – 3.74 (m, 5H), 2.70 – 2.62 (m, 4H), 2.36 (t, J =7.6 Hz, 2H), 2.00 – 1.90 (m, 6H).

[0150] Example 24

[0151] Synthesis of 3-((4-methoxyphenyl)amino)-4-((3-(piperidin-1-yl)propyl)(quinolin-6-yl)amino)cyclobut-3-ene-1,2-dione:

[0152] The synthesis method using 1-(3-bromopropyl)piperidine as a starting material is described in Example 17.

[0153] The NMR data are as follows:

[0154] 1H NMR (600 MHz, DMSO-d6) δ 8.85 (dd, J = 7.5, 1.5 Hz, 1H), 8.29 (dt,J = 7.5, 1.5 Hz, 1H), 7.97 (dd, J = 7.5, 1.6 Hz, 1H), 7.86 (d, J = 7.4 Hz,1H), 7.38 (t, J = 7.5 Hz, 1H), 6.68 – 6.64 (m, 2H), 6.59 – 6.54 (m, 2H), 3.81– 3.74 (m, 5H), 2.42 (t, J = 5.5 Hz, 4H), 2.36 (t, J = 5.6 Hz, 2H), 1.94 (tt,J = 7.6, 5.6 Hz, 2H), 1.52 (p, J = 5.7 Hz, 4H), 1.41 (q, J = 6.1 Hz, 2H).

[0155] Example 25

[0156] Synthesis of 3-((4-methoxyphenyl)amino)-4-((3-(piperidin-1-yl)propyl)(quinolin-6-yl)amino)cyclobut-3-ene-1,2-dione:

[0157] The synthesis method using 1-(3-bromopropyl)piperidine as a starting material is described in Example 17.

[0158] The NMR data are as follows:

[0159] 1H NMR (600 MHz, DMSO-d6) δ 8.85 (dd, J = 7.5, 1.5 Hz, 1H), 8.27 (dt,J = 7.5, 1.5 Hz, 1H), 8.00 (dd, J = 7.5, 1.6 Hz, 1H), 7.86 (d, J = 7.5 Hz,1H), 7.38 (t, J = 7.5 Hz, 1H), 6.69 – 6.63 (m, 2H), 6.58 – 6.53 (m, 2H), 3.81– 3.74 (m, 5H), 3.56 (t, J = 4.7 Hz, 4H), 2.39 – 2.32 (m, 6H), 1.94 (p, J =7.5 Hz, 2H).

[0160] Example 26

[0161] Synthesis of 3-((2-(2-methoxyethoxy)ethyl)(quinoline-6-yl)amino)-4-((4-methoxyphenyl)amino)cyclobut-3-ene-1,2-dione:

[0162] The synthesis method is described in Example 17, using 1-bromo-2-(2-methoxyethoxy)ethane as the starting material.

[0163] The NMR data are as follows:

[0164] 1H NMR (600 MHz, DMSO-d6) δ 8.85 (dd, J = 7.5, 1.5 Hz, 1H), 8.28 (dt,J = 7.5, 1.5 Hz, 1H), 7.98 (dd, J = 7.5, 1.6 Hz, 1H), 7.86 (d, J = 7.4 Hz,1H), 7.38 (t, J = 7.5 Hz, 1H), 7.14 (t, J = 1.5 Hz, 1H), 6.81 – 6.75 (m, 2H),6.69 – 6.64 (m, 2H), 3.79 (s, 3H), 3.66 – 3.58 (m, 4H), 3.54 (td, J = 7.0,1.2 Hz, 2H), 3.31 – 3.25 (m, 5H).

[0165] Example 27

[0166] Synthesis of 3-((4-methoxyphenyl)(methyl)amino)-4-(methyl(quinolin-6-yl)amino)cyclobut-3-ene-1,2-dione:

[0167] 1.34 g of quinoline-6-amine was dissolved in acetonitrile, and 1.846 g of iodomethane and 1.696 g of potassium iodide were added. The mixture was stirred at room temperature for 30 minutes, followed by the addition of 1.828 g of potassium carbonate. The mixture was then refluxed at 70 °C and stirred for 12 hours. When the reaction was complete, the crude product was extracted with water and ethyl acetate. The organic phase was dried over anhydrous sodium sulfate and evaporated to dryness under reduced pressure. The product was purified by silica gel column chromatography, eluting with dichloromethane:methanol (50:1) to give N-methylquinoline-6-amine.

[0168] 0.642 g of N-methylquinoline-6-amine was dissolved in methanol, and 0.642 g of 3,4-dimethoxycyclobutane-3-ene-1,2-dione was added. The mixture was refluxed at 65 °C and stirred for 24 h. When the reaction was complete, the crude product was extracted with water and ethyl acetate. The organic phase was dried over anhydrous sodium sulfate and evaporated to dryness under reduced pressure. The product was purified by silica gel column chromatography, eluting with dichloromethane:methanol (20:1) to give 3-methoxy-4-(methyl(quinoline-6-yl)amino)cyclobutane-3-ene-1,2-dione.

[0169] 0.738 g of p-methoxyaniline was dissolved in acetonitrile, and 0.846 g of iodomethane and 0.696 g of potassium iodide were added. The mixture was stirred at room temperature for 30 minutes, followed by the addition of 0.828 g of potassium carbonate. The mixture was then refluxed at 70 °C and stirred for 12 hours. When the reaction was complete, the crude product was extracted with water and ethyl acetate. The organic phase was dried over anhydrous sodium sulfate and evaporated to dryness under reduced pressure. The product was purified by silica gel column chromatography, eluting with dichloromethane:methanol (20:1) to give 4-methoxy-N-methylaniline.

[0170] 0.428 g of 4-methoxy-N-methylaniline was dissolved in methanol, and 0.428 g of 3-methoxy-4-(methyl(quinoline-6-yl)amino)cyclobut-3-ene-1,2-dione was added. The mixture was refluxed at 65 °C and stirred for 24 h. When the reaction was complete, the crude product was extracted with water and ethyl acetate. The organic phase was dried over anhydrous sodium sulfate and evaporated to dryness under reduced pressure. The product was purified by silica gel column chromatography, eluting with dichloromethane:methanol (10:1) to give 3-((4-methoxyphenyl)(methyl)amino)-4-(methyl(quinoline-6-yl)amino)cyclobut-3-ene-1,2-dione.

[0171] The NMR data are as follows:

[0172] 1H NMR (600 MHz, DMSO-d6) δ 8.85 (dd, J = 7.5, 1.6 Hz, 1H), 8.31 (dt,J = 7.4, 1.5 Hz, 1H), 7.93 (dd, J = 7.5, 1.6 Hz, 1H), 7.83 (d, J = 7.5 Hz,1H), 7.38 (t, J = 7.5 Hz, 1H), 7.25 (t, J = 1.4 Hz, 1H), 6.79 – 6.73 (m, 2H), 6.70 – 6.64 (m, 2H), 3.79 (s, 3H), 3.47 (s, 5H).

[0173] Example 28

[0174] Verification of the anti-inflammatory activity of this invention against methoxyphenylaminocyclobutenedione quinoline derivatives:

[0175] The experiment used a cell line (stable expression of P2Y14R) provided by Keygen Biotech, cultured in DMEM high-glucose medium (containing 10% fetal bovine serum). Cells were seeded in 384-well plates at 1 × 10⁶ cells per well. 4 Individual samples were cultured at 37°C, 95% oxygen, and 5% carbon dioxide for 24 hours. Before the experiment, the original culture medium was removed and replaced with serum-free DMEM, and IBMX and Ro 20-1724 were added to inhibit phosphodiesterase activity and ensure that cAMP levels were maintained at a high level. cAMP production was stimulated by adding Forskolin, and different concentrations of the target compound were added for the experiment. PPTN was used as the control group. P2Y was also added. 14 R agonist UDP-Glc. After 30 minutes, the intracellular cAMP content was detected using the cAMP GloTMAssay kit, and the IC50 value and inhibition rate were calculated accordingly; the experimental results are shown in Table 1.

[0176] Example 29

[0177] Verification of the solubility of the methoxyphenylaminocyclobutenyronylquinoline derivative of this invention: The apparent solubility of the compound in water was determined by high performance liquid chromatography (HPLC). First, the compound reference standard was accurately weighed and a series of standard solutions of known concentrations were prepared. These solutions were then injected and analyzed under selected chromatographic conditions. A standard curve was established by linear regression of peak area against concentration (R0 required). 2 The chromatographic conditions were as follows: A CTS-C18 column (250 mm × 4.6 mm, 5 μm) was used; the mobile phase was 0.1% trifluoroacetic acid aqueous solution and acetonitrile, with gradient elution; the column temperature was 35℃; the flow rate was 1.0 mL / min; the detection wavelength was set according to the UV absorption characteristics of the compound; and the injection volume was 20 μL. Subsequently, an excess of the analyte was added to pure water and the mixture was shaken at 37℃ in the dark for 72 hours to reach dissolution equilibrium. After high-speed centrifugation (10,000 rpm, 15 minutes), the supernatant was accurately measured and analyzed under the same chromatographic conditions as the standard curve, and the target peak area was recorded. Finally, the measured peak area was substituted into the standard curve equation to calculate the actual concentration of the compound in the supernatant, which is the apparent solubility of the compound in water at 37℃. Parallel experiments were set up throughout the process, and blank interference was subtracted to ensure the accuracy and reproducibility of the results; the experimental results are shown in Tables 1-4.

[0178] Table 1 Screening of in vitro anti-inflammatory activity and solubility determination of test compounds - 1

[0179] Table 2 Screening of in vitro anti-inflammatory activity and solubility determination of test compounds - 2

[0180] Table 3 Screening of in vitro anti-inflammatory activity and solubility determination of test compounds - 3

[0181] Table 4 Screening of in vitro anti-inflammatory activity and solubility determination of test compounds - 4

[0182] Previous studies have shown that while compound HDB-18 exhibits excellent receptor inhibitory activity, its extremely low solubility severely limits its drug development and in vivo pharmacodynamic studies against liver fibrosis. Literature review revealed that the poor solubility of compound HDB-18 may be due to strong hydrogen bonds formed between the secondary amides in the squamamide skeleton, resulting in a high melting point and low solubility. Therefore, this invention addresses this issue by blocking the formation of strong hydrogen bonds through amino alkylation of the squamamide skeleton and introducing polar substituents such as hydroxyl, carboxyl, and piperazine heterocyclic structures or ionizable groups, effectively improving the hydrophilicity of the molecule, thereby obtaining a candidate anti-liver fibrosis compound with both good solubility and inhibitory activity. Results show that compared with HDB-18 and the control group PPTN, the new compounds of this application exhibit significantly improved solubility with minimal impact on inhibitory activity. Among them, compound No. 14 (hereinafter named LJR-8), with R1 as N-(propyl)morpholino and R2 as a hydrogen atom, maintains strong P2Y... 14 While inhibiting the activity of R, its solubility was increased by 100 times compared to HDB-18. The inhibition rate of LJR-8 was 119.87%, IC50... 50 The value is 12.26 nM, and the solubility is 675.15 μg / mL, which is 100 times higher than HDB-18 and more than 600 times higher than PPTN.

[0183] Example 32

[0184] Verification of the therapeutic effect of the present invention on the methoxyphenylaminocyclobutenylonylquinoline derivative in an animal model of liver fibrosis:

[0185] Taking 3-((4-methoxyphenyl)(3-morpholinopropyl)amino)-4-((quinolin-6-yl)amino)cyclobut-3-ene-1,2-dione (LJR-8) as an example, we studied the pharmacodynamics of this type of compound in a mouse model of liver fibrosis.

[0186] Male C57BL / 6 mice weighing 20-25g and aged 6-8 weeks were selected and randomly divided into 6 groups. Except for the normal control group (fed a standard maintenance diet), all other groups were fed a CDA-HFD diet for the corresponding number of weeks. The mice's mental state was observed daily during the modeling period. Treatment drugs were administered according to a predetermined protocol: HDB-1 (5 mg / kg) was dissolved in phosphate-buffered saline (PBS) containing 2.5% dimethyl sulfoxide (DMSO) and 1% Tween 80 and administered intraperitoneally; LJR-8 (30, 60 mg / kg) and Resmetirom (MGIG, 5 mg / kg) were dissolved in 0.5% CMC-Na solution and administered orally. Animals were sacrificed after the modeling period, and liver tissue was collected for subsequent analysis.

[0187] The results are as follows Figure 1 As shown, after 8 weeks of CDA-HFD diet modeling, the serum ALT and AST levels of mice increased significantly, while LJR-8 could significantly downregulate the serum ALT and AST levels of mice, suggesting that liver damage was improved to some extent.

[0188] like Figure 2 The mice shown had significantly increased serum TG and TC levels after 8 weeks of CDA-HFD diet modeling, while LJR-8 significantly downregulated serum TG and TC levels, suggesting that fatty liver was improved to some extent.

[0189] like Figure 3 As shown, when mice were treated with CDA-HFD diet for 8 weeks, the hydroxyproline content in their liver tissue increased significantly. In stark contrast, after intervention with LJR-8, the hydroxyproline content in the liver tissue of mice showed a significant decrease. This phenomenon strongly suggests that collagen deposition in the mouse liver was reduced, and the degree of liver fibrosis was effectively improved.

[0190] like Figure 4 As shown, compared with the control group fed a normal diet, the mRNA expression levels of the aforementioned fibrosis-related marker genes in the liver tissue of CDF-HFD-induced liver fibrosis model mice were significantly upregulated. Oral administration of the compound LJR-8 at 60 mg / kg significantly reduced the expression of these genes, suggesting that LJR-8 may alleviate the progression of liver fibrosis by inhibiting hepatic stellate cell activation and collagen synthesis and cross-linking.

[0191] like Figure 5 and 6As shown, H&E staining results indicated that, compared with the control group, the CDA-HFD model group exhibited typical fatty degeneration characteristics in liver tissue, characterized by the accumulation of a large number of lipid droplets in hepatocytes, accompanied by disordered lobular structure and hepatocyte necrosis. After LJR-8 intervention, the above-mentioned fatty degeneration, structural damage, and necrosis were significantly improved, suggesting that all three can effectively alleviate liver tissue damage induced by metabolic stress. Masson and Sirius Red staining results further showed that the liver structure in the CDA-HFD model group was severely damaged, with continuous fibrous septa forming between the portal area and the central vein, and a large amount of collagen fiber deposition. After LJR-8 intervention, the liver tissue structural damage was significantly reduced, the formation of fibrous septa decreased, and collagen deposition significantly decreased, suggesting that all three can effectively alleviate the progression of liver fibrosis.

[0192] These results indicate that compound LJR-8 not only possesses excellent anti-fibrotic activity, but its oral efficacy also demonstrates promising development potential and clinical application prospects as a candidate drug for the treatment of liver fibrosis. When applied to the treatment of liver fibrosis, the compound of this application can significantly reduce liver inflammation, tissue structure damage, and abnormal accumulation of collagen fibers, and its oral efficacy is beneficial to improving patient compliance, thus showing great clinical application potential.

Claims

1. A p-methoxyphenylaminocyclobutenyronylquinoline derivative, characterized in that, The structure of the derivative is shown in Formula I: ； R1 or R2 is selected from alkyl groups or 5- to 6-membered heterocyclic groups containing alkyl chains with a length of 2 to 3 carbon atoms.

2. The p-methoxyphenylaminocyclobutenedionylquinoline derivative according to claim 1, characterized in that, R1 or R2 is a pentacyclic or six-membered ring without substitution, with methyl, ethyl, propyl or alkylene chains as linkers, ethyl carboxylate, propanol, methyl carboxylate, ethanol, or 2-methoxyethoxy.

3. The p-methoxyphenylaminocyclobutenedionylquinoline derivative according to claim 2, characterized in that, The five-membered ring is tetrahydropyrrole, and the six-membered ring is piperidine, morpholine, or tetrahydropyran.

4. The p-methoxyphenylaminocyclobutenylonylquinoline derivative according to claim 1, characterized in that, The derivative includes the following structure: ； 。 5. A method for preparing the p-methoxyphenylaminocyclobutenidine quinoline derivative of claim 1, characterized in that, Reaction of a compound of formula II and a compound of formula III yields the p-methoxyphenylaminocyclobutenidine quinoline derivative of formula I, wherein R1 or R2 is as described in claim 1: 。 6. The preparation method according to claim 5, characterized in that, The synthetic route is as follows: 。 7. A pharmaceutical composition, characterized in that, It comprises the derivative of claim 1 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient.

8. A derivative of claim 1 or a pharmaceutical composition of claim 7 in the preparation of P2Y 14 Application in drugs for the treatment of receptor-related diseases.

9. The application according to claim 8, characterized in that, The P2Y14 receptor-related diseases include ischemic acute kidney injury, nerve damage following subarachnoid hemorrhage, inflammatory diseases, venous thromboembolism, or liver fibrosis.

10. The application according to claim 8, characterized in that, The derivatives or their pharmaceutical compositions may be combined with anti-inflammatory and hepatoprotective drugs, drugs for treating metabolic-related fatty liver disease, targeted anti-fibrotic drugs, or renin-angiotensin system blockers.

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