A cyclobutenedione-based benzoxazole derivative, its preparation method and application

Abstract

The present invention provides a cyclobutenedione-based benzoxazole derivative, a preparation method thereof and an application thereof. Specifically, the cyclobutenedione-based benzoxazole derivative has a structure shown in formula (I). Experimental results show that the cyclobutenedione-based benzoxazole derivative provided by the present invention has good P2Y 14 receptor antagonistic activity and can be used as a therapeutic drug for preparing drugs for treating P2Y 14 receptor-related diseases.

Description

Technical Field

[0001] The present invention relates to the technical field of medicinal chemistry, and particularly relates to a preparation method and application of a cyclobutenedione-based benzoxazole derivative. Background Art

[0002] P2Y 14 receptor (P2Y 14 R) is one of the members of the P2Y receptor. When P2Y 14 R is stimulated, it can promote the release of mediators by mast cells and inflammation of renal intercalated cells, increase the hypersensitivity of microglial cells and the chemotaxis of neutrophils; and can inhibit the release of matrix metalloproteinases and tumor necrosis factor by astrocytes. P2Y 14 receptor antagonists have good innovation and application prospects in the field of drug development for related diseases such as liver fibrosis, arthritis, acute kidney injury and neuropathic pain. The activation of P2Y 14 R is closely related to the intracellular cAMP content, and cAMP can prevent the activation of the NLRP3 inflammasome. Therefore, P2Y 14 R may regulate the inflammatory response through the NLRP3 inflammasome. In chronic liver diseases, fibrosis is the main factor determining the prognosis, but there is a lack of effective anti-fibrotic treatment. The latest research [1] shows that the activation of ERK induced by P2Y 14 is the main cause of the profibrotic effect of HSCs(44).

[0003] Benzoxazole is an important pharmacophore in modern drug discovery. A large number of benzoxazole compounds have been successfully developed. The inventor's research group has only disclosed benzoxazole derivatives with various structures and used them as P2Y 14 receptor antagonists, and the inhibitory effects of these compounds need to be further improved. Moreover, the poor solubility of existing compounds in aqueous solution limits the clinical application of this type of compound. Summary of the Invention

[0004] In view of this, the present invention discloses a cyclobutenedione-based benzoxazole derivative, its preparation method and application. The prepared cyclobutenedione-based benzoxazole derivative has good antagonistic activity against the P2Y 14 receptor and activity in treating related diseases such as liver fibrosis. Cyclobutenedione-based benzoxazole is an important pharmacophore in drugs. By using the principle of activity superposition, after introducing the cyclobutenedione-based benzoxazole group into many small molecule drugs, their activities are greatly improved, and they have low toxicity, high bioavailability, good biocompatibility and efficacy. Cyclobutenedione-based benzoxazole compounds have a wide range of biological activities.

[0005] The present invention adopts the following technical solutions:

[0006] A cyclobutenedione-based benzoxazole derivative has the structure shown in formula (I): Wherein, R is a ring, preferably an alicyclic ring, an aromatic ring, a substituted aromatic ring, a heterocyclic ring or a substituted heterocyclic ring; more preferably, the R is an alicyclic ring, a benzene ring, a substituted benzene ring or a heterocyclic ring, such as a 5- to 6-membered heterocyclic ring; still more preferably, the R is cyclopropane, cyclobutane, cyclohexane, cyclopentane, benzene ring and substituted benzene ring. In the substituted aromatic ring and the substituted heterocyclic ring, the substituents are independently selected from one or more of alkyl, alkoxy, haloalkyl, and haloalkoxy, and preferably the substituents are independently selected from one or more of methyl, ethyl, methoxy, fluorine atom, chlorine atom, trifluoromethyl, and trifluoromethoxy. Preferably, R is a substituted benzene ring, and the compound has good P2Y14 receptor antagonistic activity.

[0007] In some specific embodiments of the present invention, the cyclobutenedione-based benzoxazole derivative has any of the following structures:

[0008] Formula (HDB-1) Formula (HDB-2) Formula (HDB-3)

[0009] Formula (HDB-4) Formula (HDB-5) Formula (HDB-6)

[0010] Formula (HDB-7) Formula (HDB-8) Formula (HDB-9)

[0011] Formula (HDB-10) Formula (HDB-11) Formula (HDB-12)

[0012] Formula (HDB-13) Formula (HDB-14) Formula (HDB-15)

[0013] Formula (HDB-16) Formula (HDB-17).

[0014] The present invention provides a preparation method of the above-mentioned cyclobutenedione-based benzoxazole derivative, which includes the following steps: reacting 3-(benzoxazol-6-ylamino)-4-methoxycyclobut-3-ene-1,2-dione with RNH 2React to obtain the cyclobutenedione-based benzoxazole derivative. Preferably, the reaction temperature is from room temperature to 100 °C and the time is from 5 to 60 minutes; more preferably, the reaction temperature is from 50 to 70 °C and the time is from 15 to 30 minutes.

[0015] The reaction equation of the above preparation method is as follows:

[0016]

[0017] Said R 1 and R 2 have the same ranges as above and will not be elaborated here.

[0018] The present invention discloses the application of the above cyclobutenedione-based benzoxazole derivative or a pharmaceutically acceptable salt thereof in the preparation of a therapeutic drug for P2Y 14 receptor-related diseases such as liver fibrosis and other related diseases.

[0019] The present invention discloses the application of the above cyclobutenedione-based benzoxazole derivative or a pharmaceutically acceptable salt thereof in the preparation or as a P2Y 14 receptor antagonist, or in the application in the preparation of an anti-inflammatory drug.

[0020] The present invention discloses a drug for treating P2Y 14 receptor-related diseases, with the above cyclobutenedione-based benzoxazole derivative or a pharmaceutically acceptable salt thereof as the active ingredient. It further includes excipients; the excipients can be pharmaceutically acceptable excipients.

[0021] The above drug for treating liver fibrosis provided by the present invention can also be used in combination with drugs for treating other related diseases.

[0022] Compared with the prior art, the present invention provides a cyclobutenedione-based benzoxazole derivative. Experimental results show that the cyclobutenedione-based benzoxazole derivative provided by the present invention has good activity in treating P2Y 14 receptor-related diseases such as liver fibrosis. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 is the relative inhibition rate of the P2Y 14 receptor.

[0024] Figure 2 are the serum ALT and AST levels.

[0025] Figure 3 are the results of HE staining and Masson staining. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0026] To further illustrate the present invention, the cyclobutenedione-based benzoxazole derivatives provided by the present invention, their preparation methods and applications will be described in detail below in conjunction with embodiments. The raw materials used in the present invention are commercially available products, and the specific preparation operations and performance tests are conventional techniques.

[0027] Synthesis Example

[0028] Dissolve 1.34 g (10 mmol) of benz[d]oxazol-6-amine in methanol, add 1.42 g (10 mmol) of 3,4-dimethoxycyclobut-3-ene-1,2-dione, and stir the mixture at 65 °C for 24 h. When the reaction is complete, extract the crude product with water and ethyl acetate. The organic phase is dried over anhydrous sodium sulfate and evaporated to dryness under reduced pressure. The product is purified by silica gel column chromatography, eluted with dichloromethane:methanol (50:1), to obtain 1.95 g of 3-(benz[d]oxazol-6-ylamino)-4-methoxycyclobut-3-ene-1,2-dione.

[0029] Example 1

[0030] Synthesis of 3-(benz[d]oxazol-6-ylamino)-4-(p-tolylamino)cyclobut-3-ene-1,2-dione: Dissolve 0.642 g (6 mmol) of p-toluidine in methanol, add 0.732 g (3 mmol) of 3-(benz[d]oxazol-6-ylamino)-4-methoxycyclobut-3-ene-1,2-dione, and stir the mixture at 65 °C for 24 hours. When the reaction is complete, extract the crude product with water and ethyl acetate. The organic phase is dried over anhydrous sodium sulfate and evaporated to dryness under reduced pressure. The product is purified by silica gel column chromatography, eluted with dichloromethane:methanol (100:1), to obtain 3-(benz[d]oxazol-6-ylamino)-4-(p-tolylamino)cyclobut-3-ene-1,2-dione.

[0031] The NMR data is as follows: 1 H NMR (400 MHz, DMSO-d 6 ) δ 8.27 (s, 1H), 7.65 – 7.55 (m,2H), 7.19 – 7.11 (m, 2H), 6.95 – 6.85 (m, 3H), 2.32 (d, J = 1.3 Hz, 3H); 13CNMR (101 MHz, DMSO-d6) δ 183.43, 158.24, 148.33, 143.51, 143.14, 139.51, 139.28, 137.54, 130.10, 126.97, 126.50, 124.83, 119.88, 117.66, 115.83, 105.06, 17.90。

[0032] Example 2

[0033] Synthesis of 3-(benzo[d]oxazol-6-ylamino)-4-(phenylamino)cyclobut-3-ene-1,2-dione: Using 6 mmol of aniline and 3 mmol of 3-(benzo[d]oxazol-6-ylamino)-4-methoxycyclobut-3-ene-1,2-dione as raw materials, the synthesis method refers to Example 1.

[0034] The NMR data are as follows: 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.75 (s, 2H), 8.66 (s, 1H), 8.47 (s, 2H), 8.03 (d, J = 2.1 Hz, 2H), 7.74 (d, J = 8.6 Hz, 2H), 7.29 (dd, J = 8.6, 2.2 Hz, 2H); 13 C NMR (101 MHz, DMSO-d 6 ) δ 183.00, 181.54, 169.41, 164.34, 154.33, 150.61, 137.72, 135.48, 120.97, 116.05, 101.17, 53.99, 50.55, 22.78。

[0035] Example 3

[0036] Synthesis of 3-(benzo[d]oxazol-6-ylamino)-4-(cyclobutylamino)cyclobut-3-ene-1,2-dione: Using 6 mmol of cyclobutylamine and 3 mmol of 3-(benzo[d]oxazol-6-ylamino)-4-methoxycyclobut-3-ene-1,2-dione as raw materials, the synthesis method refers to Example 1.

[0037] The NMR data are as follows: 1 H NMR (400 MHz, DMSO-d 6) δ 9.79 (s, 1H), 8.66 (s, 1H), 8.00 (s, 2H), 7.74 (d, J = 8.6 Hz, 1H), 7.30 (dd, J = 8.6, 2.2 Hz, 1H), 4.55 (q, J = 8.3 Hz, 1H), 2.37 – 2.27 (m, 2H), 2.07 (td, J = 9.5, 2.7 Hz, 2H), 1.75 – 1.61 (m, 2H). 13 C NMR (101 MHz, DMSO-d 6 ) δ 180.88, 168.71, 163.59, 154.28, 150.62, 137.83, 135.43, 120.92, 116.13, 101.21, 49.27, 32.09, 14.45。

[0038] Example 4

[0039] Synthesis of 3-(benzo[d]oxazol-6-ylamino)-4-(bicyclo[1.1.1]pentan-1-ylamino)cyclobut-3-ene-1,2-dione: Using 6 mmol of bicyclo[1.1.1]-1-pentylamine hydrochloride and 3 mmol of 3-(benzo[d]oxazol-6-ylamino)-4-methoxycyclobut-3-ene-1,2-dione as raw materials, the synthesis method is referred to Example 1.

[0040] The NMR data are as follows: 1 H NMR (400 MHz, DMSO-d 6 ) δ 10.35 (s, 1H), 9.36 (s, 1H), 8.68 (s, 1H), 8.01 (s, 1H), 7.76 (d, J = 8.5 Hz, 1H), 7.40 – 7.31 (m, 2H), 7.24 – 7.16 (m, 2H), 7.06 (t, J = 6.8 Hz, 1H), 2.37 (s, 3H). 13 C NMR (101 MHz, DMSO-d 6 ) δ 182.85, 167.08, 165.90, 154.45, 150.48, 137.41, 136.55, 135.85, 130.92, 129.10, 126.90, 125.19, 122.78, 120.88, 116.72, 101.99, 18.27。

[0041] Example 5

[0042] Synthesis of 3-(benzo[d]oxazol-6-ylamino)-4-(o-tolylamino)cyclobut-3-ene-1,2-dione: Using 6 mmol of o-toluidine and 3 mmol of 3-(benzo[d]oxazol-6-ylamino)-4-methoxycyclobut-3-ene-1,2-dione as raw materials, the synthesis method is referred to Example 1.

[0043] The NMR data are as follows: 1 H NMR (400 MHz, DMSO-d 6 ) δ 10.12 (s, 1H), 9.95 (s, 1H), 8.69 (s, 1H), 8.04 (s, 1H), 7.78 (d, J = 8.4 Hz, 1H), 7.44 (d, J = 39.7 Hz, 5H), 7.09 (s, 1H), 2.50 (s, 3H). 13 C NMR (101 MHz, DMSO-d 6 ) δ 182.23, 182.13, 166.31, 165.82, 154.54, 150.51, 138.92, 137.30, 135.92, 129.86, 123.88, 120.98, 119.06, 116.59, 101.83.

[0044] Example 6

[0045] Synthesis of 3-(benzo[d]oxazol-6-ylamino)-4-(m-tolylamino)cyclobut-3-ene-1,2-dione: Using 6 mmol of m-toluidine and 3 mmol of 3-(benzo[d]oxazol-6-ylamino)-4-methoxycyclobut-3-ene-1,2-dione as raw materials, the synthesis method is referred to Example 1.

[0046] The NMR data are as follows: 1 H NMR (400 MHz, DMSO-d 6 ) δ 10.12 (s, 1H), 9.91 (s, 1H), 8.69 (s, 1H), 8.03 (s, 1H), 7.76 (s, 1H), 7.27 (d, J = 13.8 Hz, 4H), 6.89 (s, 1H), 2.31 (s, 3H). 13 C NMR (101 MHz, DMSO-d 6) δ 182.22, 182.14, 166.34, 165.76, 154.52, 150.51, 139.23, 138.85, 137.35, 135.89, 129.69, 124.64, 120.97, 119.60, 116.57, 116.26, 101.81, 21.62。

[0047] Example 7

[0048] Synthesis of 3-(benzo[d]oxazol-6-ylamino)-4-(cyclopentylamino)cyclobut-3-ene-1,2-dione: Using 6 mmol of cyclopentylamine and 3 mmol of 3-(benzo[d]oxazol-6-ylamino)-4-methoxycyclobut-3-ene-1,2-dione as raw materials, the synthesis method is referred to Example 1.

[0049] The NMR data are as follows: 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.75 (s, 1H), 8.65 (s, 1H), 8.04 (s, 1H), 7.73 (d, J = 8.6 Hz, 2H), 7.29 (dd, J = 8.6, 2.1 Hz, 1H), 4.40 (q, J = 6.5 Hz, 1H), 2.01 – 1.93 (m, 2H), 1.75 – 1.68 (m, 2H), 1.62 – 1.53 (m, 4H). 13 C NMR (101 MHz, DMSO-d6) δ 184.24, 180.62, 169.19, 163.64, 154.24, 150.64, 137.90, 135.33, 120.93, 115.98, 101.09, 56.06, 34.21, 23.64。

[0050] Example 8

[0051] Synthesis of 3-(benzo[d]oxazol-6-ylamino)-4-(cyclohexylamino)cyclobut-3-ene-1,2-dione: Using 6 mmol of cyclohexylamine and 3 mmol of 3-(benzo[d]oxazol-6-ylamino)-4-methoxycyclobut-3-ene-1,2-dione as raw materials, the synthesis method is referred to Example 1.

[0052] The NMR data are as follows: 11H NMR (400 MHz, DMSO-d6) δ 9.78 (s, 1H), 8.65 (s, 1H), 8.04 (s, 1H), 7.73 (d, J = 8.5 Hz, 2H), 7.29 (dd, J = 8.6, 2.2 Hz, 1H), 3.87 (s, 1H), 1.94 (s, 2H), 1.73 (s, 2H), 1.57 (d, J = 12.3 Hz, 1H), 1.40 – 1.25 (m, 5H). 13 13C NMR (101 MHz, DMSO-d6) δ 184.03, 180.56, 169.01, 163.64, 154.24, 150.64, 137.90, 135.33, 120.93, 115.96, 101.08, 53.14, 34.05, 25.18, 24.48。

[0053] Example 9

[0054] Synthesis of 3-(benzo[d]oxazol-6-ylamino)-4-((4-(trifluoromethyl)phenyl)amino)cyclobut-3-ene-1,2-dione: Using 6 mmol of 4-trifluoromethylaniline and 3 mmol of 3-(benzo[d]oxazol-6-ylamino)-4-methoxycyclobut-3-ene-1,2-dione as starting materials, the synthesis method is referred to Example 1.

[0055] The NMR data are as follows: 1 1H NMR (400 MHz, DMSO-d 6 ) δ 10.31 (d, J = 3.9 Hz, 2H), 8.69 (s, 1H), 7.99 (d, J = 2.1 Hz, 1H), 7.78 (d, J = 8.6 Hz, 1H), 7.71 (d, J = 8.7 Hz, 2H), 7.65 (d, J = 8.6 Hz, 2H). 13 13C NMR (101 MHz, DMSO-d6) δ 182.98, 182.05, 166.66, 165.80, 154.62, 150.42, 142.57, 137.05, 136.14, 127.14, 126.21, 123.38, 120.96, 119.02, 116.79, 102.08。

[0056] Example 10

[0057] Synthesis of 3-(benzo[d]oxazol-6-ylamino)-4-((4-chlorophenyl)amino)cyclobut-3-ene-1,2-dione: Using 6 mmol of 4-chloroaniline and 3 mmol of 3-(benzo[d]oxazol-6-ylamino)-4-methoxycyclobut-3-ene-1,2-dione as raw materials, the synthesis method is referred to Example 1.

[0058] The NMR data are as follows: 1 H NMR (400 MHz, DMSO-d 6 ) δ 10.24 (d, J = 45.4 Hz, 2H), 8.70 (s, 1H), 8.01 (s, 1H), 7.78 (d, J = 8.5 Hz, 1H), 7.70 (s, 1H), 7.37 (dt, J = 15.5, 7.1 Hz, 3H), 7.12 (d, J = 7.5 Hz, 1H). 13 C NMR (101 MHz, DMSO-d6) δ 165.92, 154.60, 150.46, 140.54, 137.17, 131.45, 123.36, 120.97, 118.77, 117.56, 116.74, 101.99.

[0059] Example 11

[0060] Synthesis of 3-(benzo[d]oxazol-6-ylamino)-4-((4-ethylphenyl)amino)cyclobut-3-ene-1,2-dione: Using 6 mmol of 4-ethylphenyl and 3 mmol of 3-(benzo[d]oxazol-6-ylamino)-4-methoxycyclobut-3-ene-1,2-dione as raw materials, the synthesis method is referred to Example 1.

[0061] The NMR data are as follows: 1 H NMR (400 MHz, DMSO-d 6 ) δ 10.07 (s, 1H), 9.89 (s, 1H), 8.68 (s, 1H), 8.04 (s, 1H), 7.76 (s, 1H), 7.39 (d, J = 8.3 Hz, 3H), 7.22 (s, 2H), 2.57 (q, J = 9.5, 8.6 Hz, 2H), 1.17 (s, 3H). 13C NMR (101 MHz, DMSO-d6) δ 182.19, 182.00, 166.28, 165.52, 154.48, 150.51, 139.52, 137.39, 136.60, 135.84, 129.05, 120.95, 119.23, 116.51, 101.74, 27.98, 16.13。

[0062] Example 12

[0063] Synthesis of 3-(benzo[d]oxazol-6-ylamino)-4-((3-(trifluoromethyl)phenyl)amino)cyclobut-3-ene-1,2-dione: Using 6 mmol of 3-(trifluoromethyl)aniline and 3 mmol of 3-(benzo[d]oxazol-6-ylamino)-4-methoxycyclobut-3-ene-1,2-dione as starting materials, the synthesis method is referred to Example 1.

[0064] The NMR data are as follows: 1 H NMR (400 MHz, DMSO-d 6 ) δ 10.32 (s, 2H), 8.70 (s, 1H), 7.97 (d, J = 22.1 Hz, 2H), 7.77 (s, 1H), 7.63 (d, J = 23.3 Hz, 2H), 7.40 (s, 2H). 13 C NMR (101 MHz, DMSO-d6) δ 183.43, 158.24, 148.33, 143.51, 143.14, 139.51, 139.28, 137.54, 130.10, 126.97, 126.50, 124.83, 119.88, 117.66, 115.83, 105.06, 17.90。

[0065] Example 13

[0066] Synthesis of 3-(benzo[d]oxazol-6-ylamino)-4-(cyclopropylamino)cyclobut-3-ene-1,2-dione: Using 6 mmol of cyclopropylamine and 3 mmol of 3-(benzo[d]oxazol-6-ylamino)-4-methoxycyclobut-3-ene-1,2-dione as starting materials, the synthesis method is referred to Example 1.

[0067] The NMR data are as follows: 1 H NMR (400 MHz, DMSO-d 6) δ 9.69 (s, 1H), 8.65 (s, 1H), 7.98 (s, 1H), 7.73 (d, J = 8.6 Hz, 1H), 7.29 (d, J = 8.0 Hz, 1H), 3.12 (tq, J = 7.1, 3.6 Hz, 1H), 0.79 (td, J = 7.2, 4.9 Hz, 2H), 0.70 – 0.65 (m, 2H). 13 C NMR (101 MHz, DMSO-d6) δ 181.27, 170.93, 164.11, 154.27, 150.58, 137.83, 135.40, 120.86, 26.52, 7.51。

[0068] Example 14

[0069] Synthesis of 3-(benzo[d]oxazol-6-ylamino)-4-((3-chlorophenyl)amino)cyclobut-3-ene-1,2-dione: Using 6 mmol of m-chloroaniline and 3 mmol of 3-(benzo[d]oxazol-6-ylamino)-4-methoxycyclobut-3-ene-1,2-dione as starting materials, the synthesis method is referred to Example 1.

[0070] The NMR data are as follows: 1 H NMR (400 MHz, DMSO-d 6 ) δ 10.23 (d, J = 44.4 Hz, 2H), 8.70 (s, 1H), 8.01 (d, J = 2.1 Hz, 1H), 7.78 (d, J = 8.6 Hz, 1H), 7.70 (s, 1H), 7.42 – 7.33 (m, 3H), 7.12 (d, J = 7.8 Hz, 1H). 13 C NMR (101 MHz, DMSO-d6) δ 166.28, 165.92, 154.60, 150.45, 140.53, 137.16, 136.05, 134.24, 131.46, 123.37, 120.97, 118.78, 117.58, 116.76, 102.02。

[0071] Example 15

[0072] Synthesis of 3-(benzo[d]oxazol-6-ylamino)-4-((3-ethylphenyl)amino)cyclobut-3-ene-1,2-dione: Using 6 mmol of m-ethylaniline and 3 mmol of 3-(benzo[d]oxazol-6-ylamino)-4-methoxycyclobut-3-ene-1,2-dione as raw materials, the synthesis method is referred to Example 1.

[0073] The NMR data are as follows: 1 H NMR (400 MHz, DMSO-d 6 ) δ 10.04 (d, J = 65.9 Hz, 2H),8.69 (s, 1H), 8.04 (s, 1H), 7.78 (s, 1H), 7.32 (d, J = 44.9 Hz, 3H), 6.97 (s,1H), 6.66 (s, 1H), 4.40 (s, 1H), 3.77 (s, 3H), 1.23 (s, 1H). 13 C NMR (101 MHz,DMSO-d6) δ 183.43, 158.24, 148.33, 143.73, 142.08, 139.28, 138.78, 137.54,129.58, 125.81, 121.20, 119.88, 119.43, 117.66, 105.06, 28.97, 15.60.

[0074] Example 16

[0075] Synthesis of 3-(benzo[d]oxazol-6-ylamino)-4-((2-ethylphenyl)amino)cyclobut-3-ene-1,2-dione: Using 6 mmol of o-ethylaniline and 3 mmol of 3-(benzo[d]oxazol-6-ylamino)-4-methoxycyclobut-3-ene-1,2-dione as raw materials, the synthesis method is referred to Example 1.

[0076] The NMR data are as follows: 1 H NMR (400 MHz, DMSO-d 6) δ 10.49 (s, 1H), 9.49 (s, 1H), 8.67 (s, 1H), 8.04 (d, J = 2.1 Hz, 1H), 7.76 (d, J = 8.5 Hz, 1H), 7.40 (dd, J = 8.6, 2.2 Hz, 1H), 7.30 (dd, J = 7.9, 1.3 Hz, 1H), 7.25 (dd, J = 7.5, 1.6 Hz, 1H), 7.22 – 7.15 (m, 1H), 7.12 (td, J = 7.4, 1.4 Hz, 1H), 2.76 (q, J = 7.5 Hz, 2H), 1.21 (t, J = 7.5 Hz, 3H). 13 C NMR (101 MHz, DMSO-d6) δ 182.79, 182.35, 167.30, 165.89, 154.41, 150.49, 137.51, 135.80, 135.24, 128.91, 126.76, 125.65, 123.57, 120.86, 116.69, 101.92, 49.06, 24.05, 14.56。

[0077] Example 17

[0078] Synthesis of 3-(benzo[d]oxazol-6-ylamino)-4-((3-methoxyphenyl)amino)cyclobut-3-ene-1,2-dione: Using 6 mmol of m-methoxyaniline and 3 mmol of 3-(benzo[d]oxazol-6-ylamino)-4-methoxycyclobut-3-ene-1,2-dione as starting materials, the synthesis method is referred to Example 1.

[0079] The NMR data are as follows: 1 H NMR (400 MHz, DMSO-d 6 ) δ 10.07 (s, 1H), 9.89 (s, 1H), 8.67 (s, 1H), 8.03 (s, 1H), 7.77 (s, 1H), 7.30 (d, J = 20.6 Hz, 4H), 6.92 (d, J = 6.9 Hz, 1H), 2.61 (s, 3H). 1313C NMR (101 MHz, DMSO-d6) δ 182.18, 166.36, 165.71, 154.50, 150.52, 145.57, 138.89, 137.36, 135.88, 129.74, 123.50, 120.97, 118.42, 116.51, 101.76, 28.63, 15.78。

[0080] The product structural formula of the above examples is shown in Table 1.

[0081]

[0082] Example 18 P2Y 14 P2Y12R antagonistic activity test

[0083] The HEK293 cell line stably expressing human P2Y 14 receptor was purchased from Keygen Biotech. Approximately 24 hours before the assay, the cells were seeded in a 384-well plate at a density of 1×10 4 cells per well. Before the assay, the medium was discarded and replaced with serum-free medium. IBMX (500 μM) and Ro 20-1724 (100 μM) were added to inhibit the activity of PDEs. The AC agonist Forskolin (30 μM) was used to stimulate the production of cAMP in the cells. Different concentrations of cyclobutenedione benzoxazole derivatives (0.0001, 0.001, 0.01, 0.1, 1, 10, 100 nM, aqueous solution) were added separately in advance, and PPTN (CAS No.: 1160271-30-6) was used as a positive control. At the same time, 10 μM of P2Y 14 receptor agonist UDPG was added. After 30 min, the intracellular cAMP content was detected according to the instructions of the cAMP GloTM Assay kit (PROMEGA Co., Ltd, USA). The IC 50

[0084] value and the relative inhibition rate of the P2Y 14 receptor were calculated based on the cAMP content. The results are shown in Figure 1 and Table 2.

[0085]

[0086] Example 19 P2Y 14 Experiment on the new P2Y12R antagonist for alleviating liver fibrosis

[0087] Animal model experiments are conventional techniques and comply with the relevant requirements of Soochow University. Male C57BL / 6 mice, 6 - 8 weeks old, weighing 20 - 25 g, with free access to food and water, 12 h of lighting per day, and the environmental temperature of 25 ± 2 °C. The mice were randomly divided into 4 groups: sham operation control group, model control group, positive control group (magnesium isoglycyrrhizinate 20 mg / kg), and HDB-1 group (HDB-1 10 mg / kg). A mouse fibrosis model was constructed using common bile duct ligation. After laparotomy, the common bile duct was isolated and ligated twice with 5-0 silk thread. The other steps of the sham operation group were the same as those of the model group except for not ligating. On the second day after bile duct ligation, the treatment groups were intraperitoneally injected with drugs once a day, and the sham operation control group was injected with the same dose of normal saline until the end of model establishment. After 14 days, blood was collected by eye socket puncture, and the serum ALT and AST levels were measured using ALT and AST kits. The results are shown in Figure 2 , ** P <0.01, *** P <0.001, compared with the model group. The mice were sacrificed to obtain liver tissues for fixation for HE staining and Masson staining. The results are shown in Figure 3 .

[0088] Example 20 P2Y 14 In vivo pharmacokinetic study of a novel P2Y

[0089] Twelve healthy male SD rats, weighing about 220 g, were adaptively fed for 5 days with free access to food and water. They were fasted overnight before the experiment and randomly divided into 2 groups (n = 6). Compound HDB-1 was administered by gavage at 20 mg / kg; the administration volume was 0.2 mL / 100 g; and it was administered via the tail vein at 2 mg / kg; the administration volume was 0.2 mL / 100 g. Blood samples were continuously collected from the fundus venous plexus at 0 h (before dosing), 10 min, 20 min, 30 min, 45 min, 1 h, 75 min, 90 min, 2 h, 4 h, 6 h, 8 h, 12 h after gavage administration; and at 2 min, 5 min, 10 min, 30 min, 45 min, 1 h, 2 h, 4 h, 6 h, 8 h after intravenous injection into heparin-treated EP tubes. The whole blood was centrifuged at 8000 rpm for 5 min, and the plasma samples were taken and stored at -80 °C. The drug concentration in the plasma was analyzed and determined by LC-MS / MS (Shimadzu LCMS-8030), and the pharmacokinetic parameters were analyzed using the WinNonlin Professional V6.3 non-compartment model. The results are shown in Table 3, indicating that HDB1 has good metabolic stability and oral bioavailability.

[0090]

[0091] As can be seen from the above embodiments, the cyclobutenedione-based derivative provided by the present invention has the structure shown in formula (I). The experimental results show that the cyclobutenedione-based derivative provided by the present invention has good P2Y 14 receptor antagonistic activity, in vivo anti-inflammatory activity and pharmacokinetic properties, and can be used as an application for preparing drugs for treating P2Y 14 receptor-related inflammatory diseases.

[0092] The description of the above embodiments is only used to help understand the method and its core idea of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and modifications can be made to the present invention, and these improvements and modifications also fall within the protection scope of the claims of the present invention.

Claims

1. A cyclobutenedione-based benzoxazole derivative having the structure shown in formula (I): ； Wherein, R is cyclopropane, cyclobutane, cyclohexane, cyclopentane, benzene ring, substituted benzene ring or substituted 5-6 membered heterocycle; in the substituted benzene ring and the substituted 5-6 membered heterocycle, the substituents are independently selected from one or more of methyl, ethyl, methoxy, fluorine atom, chlorine atom, trifluoromethyl, trifluoromethoxy.

2. A method for preparing the cyclobutenedione-based benzoxazole derivative according to claim 1, Characterized in that, It comprises the following steps: React 3-(benzo[d]oxazol-6-ylamino)-4-methoxycyclobut-3-ene-1,2-dione with RNH 2 to obtain the cyclobutenedione-based benzoxazole derivative.

3. According to the method for preparing the cyclobutenedione-based benzoxazole derivative according to claim 2, Characterized in that, The reaction temperature is room temperature to 100 °C and the time is 5 to 60 minutes.

4. Use of the cyclobutenedione-based benzoxazole derivative or a pharmaceutically acceptable salt thereof according to claim 1 in the preparation of a therapeutic drug for treating P2Y 14 receptor-related diseases.

5. Use of the cyclobutenedione-based benzoxazole derivative or a pharmaceutically acceptable salt thereof according to claim 1 in the preparation of, or as, a P2Y 14 receptor antagonist.

6. Use of the cyclobutenedione-based benzoxazole derivative according to claim 1 or a pharmaceutically acceptable salt thereof in the preparation of an anti-inflammatory drug.

7. A drug for treating P2Y 14 receptor-related diseases Characterized in that, The cyclobutenedione-based benzoxazole derivative according to claim 1 or a pharmaceutically acceptable salt thereof is used as an active ingredient.

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