Tryptanthrin compound as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of organic compound preparation, and discloses a tryptanthrin compound as well as a preparation method and application thereof. The preparation method of the tryptanthrin compound disclosed by the invention comprises the step of enabling an indole-2, 3-diketone derivative, 2H-thieno [3, 2-D] [1, 3] oxazine-2, 4 (1H)-diketone, triethylamine and methylbenzene to react, so as to obtain the tryptanthrin compound. The obtained tryptanthrin compound is novel in structure, shows ulcerative colitis resisting activity and has the potential of being developed into ulcerative colitis resisting drugs.

Description

Technical Field

[0001] The present invention relates to the technical field of organic compound preparation, in particular to a tryptanthrin compound and a preparation method and application thereof. Background Art

[0002] Ulcerative colitis (UC) is a chronic, nonspecific inflammatory bowel disease that primarily affects the mucosal and submucosa layers of the colon and rectum. Typical symptoms include recurrent diarrhea, bloody and mucus-purulent stools, abdominal pain, tenesmus (a feeling of incomplete defecation), and weight loss. Severe cases may be accompanied by systemic symptoms such as fever and anemia. Lesions typically begin in the rectum and extend continuously toward the proximal colon. The extent of inflammation correlates with disease severity. The etiology is currently unclear but may be related to a multifactorial interaction, including genetic susceptibility, abnormal immune system activation, intestinal dysbiosis, and environmental factors (such as diet and smoking). Treatment aims to control inflammation, alleviate symptoms, and reduce recurrences. Mild to moderate cases are commonly treated with aminosalicylic acids (such as mesalamine). Moderate to severe or refractory cases require treatment with glucocorticoids, immunosuppressants (such as azathioprine), or biologics (such as anti-TNF-α monoclonal antibodies). The development of new anti-UC drugs remains an urgent challenge in this field.

[0003] Tryptanthrin is a natural indole alkaloid widely found in medicinal plants such as Radix Isatidis. In recent years, it has become a research hotspot for the treatment of ulcerative colitis due to its significant anti-inflammatory and immunomodulatory activities. Studies have shown that tryptanthrin can inhibit intestinal inflammatory responses through multiple targets: (1) blocking key signaling pathways such as NF-κB and STAT3, reducing the expression of pro-inflammatory factors such as TNF-α, IL-6, and IL-1β; (2) regulating the balance of Th17 / Treg cells and inhibiting excessive immune responses; (3) enhancing intestinal barrier function and reducing oxidative stress damage. Animal experiments have shown that tryptanthrin can significantly alleviate the symptoms of dextran sulfate sodium (DSS)-induced colitis in mice, reduce the disease activity index and tissue pathology score. To improve efficacy and reduce toxicity, researchers have modified the structure of tryptanthrin and developed a series of derivatives. For example, the introduction of sulfonic acid groups can enhance water solubility, fluorinated derivatives show stronger NF-κB inhibitory activity, and C-6 hydroxyl substituents show better antioxidant capacity. Currently, tryptanthrin provides a potential direction for the development of anti-ulcerative colitis drugs. The development of anti-ulcerative colitis drugs based on tryptanthrin is of great significance for the development of new drug treatments.

[0004] Therefore, it is of great significance to study and obtain a tryptanthrin compound with novel structure and anti-ulcerative colitis activity as well as its preparation method and application. Summary of the Invention

[0005] In view of this, the present invention provides a tryptanthrin compound and a preparation method and application thereof, the purpose of which is to solve the problem that existing tryptanthrin compounds have low efficacy and high toxicity.

[0006] In order to achieve the above object, the present invention adopts the following technical solutions:

[0007] The present invention provides a tryptanthrin compound, the structural formula of the tryptanthrin compound is as follows:

[0008]

[0009] Said R1, R2 and R3 are independently hydrogen, halogen, methoxy, methyl, nitroso or trifluoromethoxy.

[0010] Preferably, the structural formula of the tryptanthrin compound includes:

[0011]

[0012]

[0013] The present invention also provides a method for preparing the tryptanthrin compound, comprising the following steps:

[0014] Tryptanthrin compounds are obtained by reacting an indole-2,3-dione derivative, 2H-thieno[3,2-D][1,3]oxazine-2,4(1H)-dione, triethylamine and toluene.

[0015] Preferably, the structural formula of the indole-2,3-dione derivative is:

[0016]

[0017] Said R1, R2 and R3 are independently hydrogen, halogen, methoxy, methyl, nitroso or trifluoromethoxy.

[0018] Preferably, the usage ratio of the indole-2,3-dione derivative, 2H-thieno[3,2-D][1,3]oxazine-2,4(1H)-dione, triethylamine and toluene is 0.8-1.2 mmol: 0.8-1.2 mmol: 0.8-1.2 mL: 8-12 mL.

[0019] Preferably, the reaction time is 4 to 8 hours, and the reaction temperature is 80 to 100°C.

[0020] The present invention also provides use of the tryptanthrin compound in the preparation of anti-inflammatory drugs.

[0021] It can be seen from the above technical solutions that compared with the prior art, the present invention has the following beneficial effects:

[0022] The invention prepares a tryptanthrin compound, which has a novel structure, exhibits anti-ulcerative colitis activity and has the potential to be developed into an anti-ulcerative colitis drug. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are merely embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on the provided drawings without paying any creative work.

[0024] Figure 1 This is a graph showing the NO secretion induced by bacterial lipopolysaccharide in response to the compounds obtained in Examples 1 to 16;

[0025] Figure 2 This is a graph showing the effect of compound 21 obtained in Example 12 on cell viability;

[0026] Figure 3 This is a graph showing the effect of compound 21 obtained in Example 12 on the disease activity index;

[0027] Figure 4 The effect of compound 21 obtained in Example 12 on the body weight of mice with ulcerative colitis;

[0028] Figure 5 Effect of compound 21 obtained in Example 12 on colon length in mice with ulcerative colitis;

[0029] Figure 6 This is a statistical graph showing the effect of compound 21 obtained in Example 12 on the colon length of mice with ulcerative colitis. DETAILED DESCRIPTION

[0030] The present invention provides a tryptanthrin compound, the structural formula of the tryptanthrin compound is as follows:

[0031]

[0032] Said R1, R2 and R3 are independently hydrogen, halogen, methoxy, methyl, nitroso or trifluoromethoxy.

[0033] In the present invention, among the tryptanthrin compounds, the structural formula of tryptanthrin is as follows:

[0034]

[0035] In the present invention, the structural formula of the tryptanthrin compound preferably includes:

[0036]

[0037]

[0038] The present invention also provides a method for preparing the tryptanthrin compound, comprising the following steps:

[0039] Tryptanthrin compounds are obtained by reacting an indole-2,3-dione derivative, 2H-thieno[3,2-D][1,3]oxazine-2,4(1H)-dione, triethylamine and toluene.

[0040] In the present invention, the structural formula of the indole-2,3-dione derivative is preferably:

[0041]

[0042] Said R1, R2 and R3 are preferably independently hydrogen, halogen, methoxy, methyl, nitroso or trifluoromethoxy.

[0043] In the present invention, the usage ratio of the indole-2,3-dione derivative, 2H-thieno[3,2-D][1,3]oxazine-2,4(1H)-dione, triethylamine and toluene is preferably 0.8-1.2 mmol: 0.8-1.2 mmol: 0.8-1.2 mL: 8-12 mL, more preferably 0.9-1.1 mmol: 0.9-1.1 mmol: 0.9-1.1 mL: 9-11 mL, and more preferably 1 mmol: 1 mmol: 1 mL: 10 mL.

[0044] In the present invention, the reaction is a condensation reaction, and the synthetic route of the reaction is:

[0045]

[0046] In the present invention, the reaction time is preferably 4 to 8 hours, more preferably 5 to 7 hours, more preferably 6 hours, and the reaction temperature is preferably 80 to 100°C, more preferably 85 to 95°C, more preferably 90 to 92°C.

[0047] In the present invention, the reaction is followed by cooling, filtering, washing and purification;

[0048] The cooling temperature is preferably 20-30°C, more preferably 22-28°C, and more preferably 25-26°C;

[0049] The washing reagent is preferably methanol, and the number of washing times is preferably 3 to 6 times;

[0050] The purification is preferably performed by silica gel column chromatography, wherein the eluent of the silica gel column chromatography is a n-hexane-ethyl acetate system, and the volume ratio of n-hexane to ethyl acetate is preferably 3 to 5:1, more preferably 4:1.

[0051] The present invention also provides use of the tryptanthrin compound in the preparation of anti-inflammatory drugs.

[0052] The technical solutions provided by the present invention are described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0053] In the embodiment, the structural formula of the indole-2,3-dione derivative is:

[0054]

[0055] Said R1, R2 and R3 are independently hydrogen, halogen, methoxy, methyl, nitroso or trifluoromethoxy.

[0056] Example 1

[0057] Compound 2a:

[0058]

[0059] Preparation of compound 2a:

[0060] 1 mmol of indole-2,3-dione (R1, R2, and R3 are all hydrogen), 1 mmol of 2H-thieno[3,2-D][1,3]oxazine-2,4(1H)-dione, 1 mL of triethylamine, and 10 mL of toluene were condensed at 90°C for 6 h to obtain a crude product;

[0061] The resulting product was cooled to 25°C and filtered. The filtered solid was washed three times with methanol and then purified by silica gel column chromatography (eluent: n-hexane-ethyl acetate, n-hexane:ethyl acetate ratio: 4:1) to obtain compound 2a as a yellow solid. The calculated yield was 70%.

[0062] After testing: 1 H NMR (400MHz, DMSO-d6) δ8.43(d,J=8.0Hz,1H),8.33(d,J=5.2Hz,1H),7.89–7.80(m,2H),7.64(d,J=5.2Hz,1H),7.47(t,J=7.5Hz,1H)ppm; 13C NMR (101MHz, DMSO-d6) δ182.11,155.50,154.23,147.08,145.97,137.65,136.62,127.15,126.66,126.42,124.91,122.51,117.11ppm.

[0063] Example 2

[0064] Compound 2b:

[0065]

[0066] Preparation of compound 2b:

[0067] 1 mmol of 5-fluoroindole-2,3-dione (R2 is fluorine, R1 and R3 are both hydrogen), 1 mmol of 2H-thieno[3,2-D][1,3]oxazine-2,4(1H)-dione, 1 mL of triethylamine and 10 mL of toluene were condensed at 90°C for 6 h to obtain a crude product;

[0068] The resulting product was cooled to 25°C and filtered. The filtered solid was washed three times with methanol and then purified by silica gel column chromatography (eluent: n-hexane-ethyl acetate, n-hexane:ethyl acetate ratio: 4:1) to obtain compound 2b as a yellow solid. The calculated yield was 56%.

[0069] After testing: 1 H NMR (400MHz, DMSO-d6) δ8.44 (dd, J=8.8, 4.2Hz, 1H), 8.34 (d, J=5.2Hz, 1H), 7.7 8(dd,J=7.1,2.8Hz,1H),7.69(td,J=9.0,2.8Hz,1H),7.65(d,J=5.3Hz,1H)ppm; 13 C NMR(101MHz,DMSO-d6)δ181.22,159.18,155.43,154.05,142.26,136.76,12 6.66,126.37,124.39,123.81,123.57,118.90,118.82,112.00,111.76ppm.

[0070] Example 3

[0071] Compound 2c:

[0072]

[0073] Preparation of compound 2c:

[0074] 1 mmol of 5-chloroindole-2,3-dione (R2 is chlorine, R1 and R3 are both hydrogen), 1 mmol of 2H-thieno[3,2-D][1,3]oxazine-2,4(1H)-dione, 1 mL of triethylamine and 10 mL of toluene were condensed at 90°C for 6 h to obtain a crude product;

[0075] The resulting product was cooled to 25°C and filtered. The filtered solid was washed three times with methanol and then purified by silica gel column chromatography (eluent: n-hexane-ethyl acetate, n-hexane:ethyl acetate ratio: 4:1) to obtain compound 2c as a yellow solid. The calculated yield was 69%.

[0076] After testing: 1 H NMR (400MHz, DMSO-d6) δ8.39(d,J=8.5Hz,1H),8.32(d,J=5.2Hz,1H),7.92(d,J=2.3Hz,1H),7.86(dd,J=8.5,2.3Hz,1H),7.63(d,J=5.2Hz,1H)ppm; 13 C NMR (101MHz, DMSO-d6) δ181.00,155.53,154.13,147.05,144.44,136.97,136.73,131.48,126.74,126.31,124.52,124.34,118.70ppm.

[0077] Example 4

[0078] Compound 2d:

[0079]

[0080] Preparation of compound 2d:

[0081] 1 mmol of 5-bromoindole-2,3-dione (R2 is bromine, R1 and R3 are both hydrogen), 1 mmol of 2H-thieno[3,2-D][1,3]oxazine-2,4(1H)-dione, 1 mL of triethylamine and 10 mL of toluene were condensed at 90°C for 6 h to obtain a crude product;

[0082] The resulting product was cooled to 25°C and filtered. The filtered solid was washed three times with methanol and then purified by silica gel column chromatography (eluent: n-hexane-ethyl acetate, n-hexane:ethyl acetate ratio: 4:1) to obtain Compound 2d as a yellow solid. The calculated yield was 85%.

[0083] After testing: 1H NMR (400MHz, DMSO-d6) δ8.39–8.33(m,2H),8.07–7.99(m,2H),7.66(d,J=5.2Hz,1H)ppm; 13 C NMR (101MHz, DMSO-d6) δ180.84,155.51,154.07,146.81,144.77,139.57,136.93,127.27,126.70,126.27,124.53,119.37,118.98ppm.

[0084] Example 5

[0085] Compound 2e:

[0086]

[0087] Preparation of compound 2e:

[0088] 1 mmol of 5-iodoindole-2,3-dione (R2 is iodine, R1 and R3 are both hydrogen), 1 mmol of 2H-thieno[3,2-D][1,3]oxazine-2,4(1H)-dione, 1 mL of triethylamine and 10 mL of toluene were condensed at 90°C for 6 h to obtain a crude product;

[0089] The resulting product was cooled to 25°C and filtered. The filtered solid was washed three times with methanol and then purified by silica gel column chromatography (eluent: n-hexane-ethyl acetate, n-hexane:ethyl acetate ratio: 4:1) to obtain compound 2e as a yellow solid. The calculated yield was 81%.

[0090] After testing: 1 H NMR (400MHz, DMSO-d6) δ8.35(d,J=5.3Hz,1H),8.22(d,J=8.3Hz,1H),8.19(d,J=1.8Hz,1H),8.16(t,J=2.3Hz,1H),7.65(d,J=5.2Hz,1H)ppm; 13 C NMR (101MHz, DMSO-d6) δ180.82,155.53,154.08,146.52,145.43,145.23,136.86,132.75,126.68,126.28,124.51,119.09,91.71ppm.

[0091] Example 6

[0092] Compound 2f:

[0093]

[0094] Preparation of compound 2f:

[0095] 1 mmol of 6-chloroindole-2,3-dione (R3 is chlorine, R1 and R2 are both hydrogen), 1 mmol of 2H-thieno[3,2-D][1,3]oxazine-2,4(1H)-dione, 1 mL of triethylamine and 10 mL of toluene were condensed at 90°C for 6 h to obtain a crude product;

[0096] The resulting product was cooled to 25°C and filtered. The filtered solid was washed three times with methanol and then purified by silica gel column chromatography (eluent: n-hexane-ethyl acetate, n-hexane:ethyl acetate ratio: 4:1) to obtain Compound 2f as a yellow solid. The calculated yield was 88%.

[0097] After testing: 1 H NMR (400MHz, DMSO-d6) δ8.39(d,J=1.9Hz,1H),8.33(d,J=5.2Hz,1H),7.88(d,J=8.0Hz,1H),7.64(d,J=5.2Hz,1H),7.53(dd,J=8.1,1.9Hz,1H)ppm; 13 C NMR (101MHz, DMSO-d6) δ181.33,155.98,154.80,147.57,147.12,141.87,137.56,127.63,127.20,126.88,126.54,121.97,117.43ppm.

[0098] Example 7

[0099] Compound 2g:

[0100]

[0101] Preparation of compound 2g:

[0102] 1 mmol of 4-chloroindole-2,3-dione (R1 is chlorine, R2 and R3 are both hydrogen), 1 mmol of 2H-thieno[3,2-D][1,3]oxazine-2,4(1H)-dione, 1 mL of triethylamine and 10 mL of toluene were condensed at 90°C for 6 h to obtain a crude product;

[0103] The resulting product was cooled to 25°C and filtered. The filtered solid was washed three times with methanol and then purified by silica gel column chromatography (eluent: n-hexane-ethyl acetate system, n-hexane:ethyl acetate volume ratio of 4:1) to obtain 2 g of the compound as a yellow solid. The calculated yield was 46%.

[0104] After testing:1 H NMR (400MHz, DMSO-d6) δ8.49(d,J=8.0Hz,1H),8.41(d,J=5.3Hz,1H),7.88(t,J=8.1Hz,1H),7.72(d,J=5.2Hz,1H),7.57(d,J=8.2Hz,1H)ppm; 13 C NMR (101MHz, DMSO-d6) δ179.22,155.52,154.13,147.18,146.67,138.36,137.00,131.48,128.29,126.69,126.15,119.39,115.83ppm.

[0105] Example 8

[0106] Compound 2h:

[0107]

[0108] Preparation of compound 2h:

[0109] 1 mmol of 6-bromoindole-2,3-dione (R3 is bromine, R1 and R2 are both hydrogen), 1 mmol of 2H-thieno[3,2-D][1,3]oxazine-2,4(1H)-dione, 1 mL of triethylamine and 10 mL of toluene were condensed at 90°C for 6 h to obtain a crude product;

[0110] The resulting product was cooled to 25°C and filtered. The filtered solid was washed three times with methanol and then purified by silica gel column chromatography (eluent: n-hexane-ethyl acetate, n-hexane:ethyl acetate ratio: 4:1) to obtain Compound 2h as a yellow solid. The calculated yield was 52%.

[0111] After testing: 1 H NMR (400MHz, DMSO-d6) δ8.59(d,J=1.7Hz,1H),8.37(d,J=5.2Hz,1H),7.82(d,J=8.1Hz,1H),7.71(dd,J=8.1,1.7Hz,1H),7.67(d,J=5.2Hz,1H)ppm; 13 C NMR (101MHz, DMSO-d6) δ180.84,155.51,154.07,146.81,144.77,139.57,136.93,127.27,126.70,126.27,124.53,119.37,118.98ppm.

[0112] Example 9

[0113] Compound 2i:

[0114]

[0115] Preparation of compound 2i:

[0116] 1 mmol of 5-methylindole-2,3-dione (R2 is methyl, R1 and R3 are both hydrogen), 1 mmol of 2H-thieno[3,2-D][1,3]oxazine-2,4(1H)-dione, 1 mL of triethylamine and 10 mL of toluene were condensed at 90°C for 6 h to obtain a crude product;

[0117] The resulting product was cooled to 25°C and filtered. The filtered solid was washed three times with methanol and then purified by silica gel column chromatography (eluent: n-hexane-ethyl acetate, n-hexane:ethyl acetate ratio: 4:1) to obtain Compound 2i as a yellow solid. The calculated yield was 85%.

[0118] After testing: 1 H NMR (400MHz, DMSO-d6) δ8.32(d,J=5.3Hz,1H),8.29(d,J=8.1Hz,1H),7.68(d,J=1.8Hz,1H),7.66–7.61(m,2H),2.40(s,3H)ppm; 13 C NMR (101MHz, DMSO-d6) δ182.15,155.46,154.03,147.17,143.88,137.92,136.93,136.43,126.60,126.45,124.99,122.53,116.85,20.41ppm.

[0119] Example 10

[0120] Compound 2j:

[0121]

[0122] Preparation of compound 2j:

[0123] 1 mmol of 5-methoxyindole-2,3-dione (R2 is methoxy, R1 and R3 are both hydrogen), 1 mmol of 2H-thieno[3,2-D][1,3]oxazine-2,4(1H)-dione, 1 mL of triethylamine and 10 mL of toluene were condensed at 90°C for 6 h to obtain a crude product;

[0124] The resulting product was cooled to 25°C and filtered. The filtered solid was washed three times with methanol and then purified by silica gel column chromatography (eluent: n-hexane-ethyl acetate, n-hexane:ethyl acetate ratio: 4:1) to obtain Compound 2j as a yellow solid. The calculated yield was 87%.

[0125] After testing: 1 H NMR (400MHz, DMSO-d6) δ8.34–8.28(m,2H),7.62(d,J=5.3Hz,1H),7.41–7.36(m,2H),3.86(s,3H)ppm; 13 C NMR (101MHz, DMSO-d6) δ182.00,158.15,155.36,153.86,147.32,139.71,136.29,126.58,126.53,123.65,123.26,118.29,108.97,56.04ppm.

[0126] Example 11

[0127] Compound 2k:

[0128]

[0129] Preparation of compound 2k:

[0130] 1 mmol of 5-trifluoromethoxyindole-2,3-dione (R2 is trifluoromethoxy, R1 and R3 are both hydrogen), 1 mmol of 2H-thieno[3,2-D][1,3]oxazine-2,4(1H)-dione, 1 mL of triethylamine and 10 mL of toluene were condensed at 90°C for 6 h to obtain a crude product;

[0131] The resulting product was cooled to 25°C and filtered. The filtered solid was washed three times with methanol and then purified by silica gel column chromatography (eluent: n-hexane-ethyl acetate, n-hexane:ethyl acetate ratio: 4:1) to obtain Compound 2k as a yellow solid. The calculated yield was 65%.

[0132] After testing: 1 H NMR (400MHz, DMSO-d6) δ8.49(d,J=8.7Hz,1H),8.33(d,J=5.2Hz,1H),7.89(d,J=2.5Hz,1H),7.84(dd,J=8.6,2.6Hz,1H),7.64(d,J=5.2Hz,1H)ppm; 13C NMR (101MHz, DMSO-d6) δ180.87,155.46,154.08,17.23,146.44,144.44,136.99,129.90,126.70,126.23,124.28,121.28,118.74,117.84ppm.

[0133] Example 12

[0134] Compound 21:

[0135]

[0136] Preparation of compound 21:

[0137] 1 mmol of 5-nitroindole-2,3-dione (R2 is a nitroso group, and R1 and R3 are both hydrogen), 1 mmol of 2H-thieno[3,2-D][1,3]oxazine-2,4(1H)-dione, 1 mL of triethylamine, and 10 mL of toluene were condensed at 90°C for 6 h to obtain a crude product;

[0138] The resulting product was cooled to 25°C and filtered. The filtered solid was washed three times with methanol and then purified by silica gel column chromatography (eluent: n-hexane-ethyl acetate system, n-hexane:ethyl acetate volume ratio of 4:1) to obtain Compound 21 as a yellow solid. The calculated yield was 61%.

[0139] After testing: 1 H NMR (400MHz, DMSO-d6) δ8.65(dd,J=8.8,2.4Hz,1H),8.58(d,J=8.8Hz,1H),8.48(d,J=2.3Hz,1H),8.35(d,J=5.2Hz,1H),7.65(d,J=5.2Hz,1H)ppm; 13 CNMR(101MHz,DMSO-d6)δ180.34,155.35,154.18,149.21,147.38,145.80,137.58,132.51,126.81,126.03,123.44,119.55,117.73ppm.

[0140] Example 13

[0141] Compound 2m:

[0142]

[0143] Preparation of compound 2m:

[0144] 1 mmol of 4,5-dichloroindole-2,3-dione (R1 and R2 are both chlorine, R3 is hydrogen), 1 mmol of 2H-thieno[3,2-D][1,3]oxazine-2,4(1H)-dione, 1 mL of triethylamine, and 10 mL of toluene were condensed at 90°C for 6 h to obtain a crude product;

[0145] The resulting product was cooled to 25°C and filtered. The filtered solid was washed three times with methanol and then purified by silica gel column chromatography (eluent: n-hexane-ethyl acetate, n-hexane:ethyl acetate ratio: 4:1) to obtain Compound 2m as a yellow solid. The calculated yield was 54%.

[0146] After testing: 1 H NMR (400MHz, DMSO-d6) δ8.57(s,1H),8.38(d,J=5.2Hz,1H),8.22(s,1H),7.68(d,J=5.3Hz,1H)ppm; 13 C NMR (101MHz, DMSO-d6) δ179.89,155.49,153.99,146.82,144.41,138.88,137.28,129.84,126.71,126.36,125.93,122.92,118.53ppm.

[0147] Example 14

[0148] Compound 2n:

[0149]

[0150] Preparation of compound 2n:

[0151] 1 mmol of 5,6-dichloroindole-2,3-dione (R2 and R3 are both chlorine, R1 is hydrogen), 1 mmol of 2H-thieno[3,2-D][1,3]oxazine-2,4(1H)-dione, 1 mL of triethylamine and 10 mL of toluene were condensed at 90°C for 6 h to obtain a crude product;

[0152] The resulting product was cooled to 25°C and filtered. The filtered solid was washed three times with methanol and then purified by silica gel column chromatography (eluent: n-hexane-ethyl acetate, n-hexane:ethyl acetate ratio: 4:1) to obtain compound 2n as a yellow solid. The calculated yield was 67%.

[0153] After testing: 1H NMR (400MHz, DMSO-d6) δ8.46–8.34(m,2H),8.07(d,J=8.6Hz,1H),7.67(d,J=5.2Hz,1H)ppm; 13 C NMR (101MHz, DMSO-d6) δ178.39,155.52,154.00,146.58,145.55,137.70,137.28,130.34,129.46,126.72,126.02,121.17,116.78ppm.

[0154] Example 15

[0155] Compound 2o:

[0156]

[0157] Preparation of compound 2o:

[0158] 1 mmol of 4-chloro-5-fluoroindole-2,3-dione (R1 is chlorine, R2 is fluorine, and R3 is hydrogen), 1 mmol of 2H-thieno[3,2-D][1,3]oxazine-2,4(1H)-dione, 1 mL of triethylamine, and 10 mL of toluene were condensed at 90°C for 6 h to obtain a crude product;

[0159] The resulting product was cooled to 25°C and filtered. The filtered solid was washed three times with methanol and then purified by silica gel column chromatography (eluent: n-hexane-ethyl acetate, n-hexane:ethyl acetate ratio: 4:1) to obtain compound 2o as a yellow solid. The calculated yield was 69%.

[0160] After testing: 1 H NMR (400MHz, DMSO-d6) δ8.42(dd,J=8.8,3.8Hz,1H),8.36(d,J=5.3Hz,1H),7.86(t,J=9.2Hz,1H),7.67(d,J=5.2Hz,1H)ppm; 13 C NMR(101MHz,DMSO-d6)δ178.63,157.07,155.40,154.62,153.98,146.85,142.92,13 7.07,126.64,126.10,123.98,123.75,120.94,118.59,118.38,116.88,116.80ppm.

[0161] Example 16

[0162] Compound 2p:

[0163]

[0164] Preparation of compound 2p:

[0165] 1 mmol of 5-fluoro-6-chloroindole-2,3-dione (R2 is fluorine, R3 is chlorine, and R1 is hydrogen), 1 mmol of 2H-thieno[3,2-D][1,3]oxazine-2,4(1H)-dione, 1 mL of triethylamine, and 10 mL of toluene were condensed at 90°C for 6 h to obtain a crude product;

[0166] The resulting product was cooled to 25°C and filtered. The filtered solid was washed three times with methanol and then purified by silica gel column chromatography (eluent: n-hexane-ethyl acetate, n-hexane:ethyl acetate ratio: 4:1) to obtain Compound 2p as a yellow solid. The calculated yield was 63%.

[0167] After testing: 1 H NMR (400MHz, DMSO-d6) δ8.50(d,J=6.1Hz,1H),8.34(d,J=5.2Hz,1H),8.04(d,J=7.5Hz,1H),7.64(d,J=5.2Hz,1H)ppm; 13 C NMR(101MHz,DMSO-d6)δ180.69,157.65,155.94,155.18,154.47,147.52,142.5 6,142.53,137.68,127.20,126.51,123.53,123.46,119.15,113.86,113.62ppm.

[0168] The NO secretion amount of the compounds obtained in Examples 1 to 16 was tested respectively:

[0169] RAW264.7 cells were diluted with complete culture medium to 2.0 × 10 cells per well. 5 Cells were seeded in a 96-well plate. After the cells adhered, the supernatant was gently aspirated and 90 μL of complete medium containing compounds 1-16 (5 μM concentration) was added to each well. After 2 hours, 10 μL of 1 μg / mL bacterial lipopolysaccharide (LPS) was added to each well. Culture was continued for 20 hours, and the supernatant was collected.

[0170] The NO content was determined using the Griess method: 50 μL of cell supernatant was taken from each sample well, 50 μL of sulfanilamide solution was added, and the reaction was in the dark for 10 minutes. Subsequently, 50 μL of N-(1-naphthyl)ethylenediamine solution was added, and the reaction was continued in the dark for 10 minutes. Finally, the absorbance of each well was measured at a wavelength of 562 nm on a microplate reader. The results were as follows: Figure 1 shown.

[0171] The NO secretion induced by bacterial lipopolysaccharide induced by the compounds obtained in Examples 1 to 16 is shown in the figure Figure 1 As shown. Figure 1 As shown, compounds 2c, 2d, 2e, 2h, 2j, 2k, and 2l exhibited nitric oxide (NO) inhibition (relative to the inflammatory model group, i.e., the LPS group in the figure), demonstrating anti-inflammatory activity and potential as anti-inflammatory drugs. In particular, 2l exhibited the most significant inhibition of NO secretion at the same molar concentration, demonstrating significant anti-inflammatory activity and potential for further development.

[0172] The MTT cell viability test was performed on compound 21 obtained in Example 12:

[0173] The specific steps are as follows: RAW264.7 cells were prepared with complete culture medium to a density of 1.25×10 5 A single cell suspension of 100 μL / mL was inoculated into each well of a 96-well plate. After the cells were cultured under appropriate conditions for 24 hours, the original culture medium was discarded and 100 μL of drug solution of different concentrations was added. The drug to be tested was set to six concentration gradients: 100 μM, 50 μM, 25 μM, 12.5 μM, 6.25 μM and 3.125 μM (the drug stock solution was configured to 1 mmol / L using biological-grade DMSO, and then gradiently diluted with culture medium when used). A blank group and a control group were also set up. After continuing to culture for 20 hours, 10 μL of MTT solution was added to each well, incubated for 4 hours, and then DMSO was added and shaken for 1 minute. Finally, the absorbance value (A) of each well at a wavelength of 490 nm was measured using an enzyme reader, and the cell viability was calculated according to the following formula. The results are as follows Figure 2 shown.

[0174] Cell viability (%) = (A treatment group - A control group) / (A control group - A blank group) × 100%.

[0175] The following performance tests were performed on the compound 21 obtained in Example 12:

[0176] Animal grouping and drug administration: Based on the test results of MTT and NO secretion, tryptanthrin and compound 21 with good anti-inflammatory activity and low cytotoxicity were selected for the experiment.

[0177] The experimental animals were divided into the following groups: control group, model group, positive control group with sulfasalazine added at a dose of 125 mg / kg, tryptanthrin group (TRYP) at a dose of 78 mg / kg, and compound 21 group at a dose of 108 mg / kg.

[0178] Modeling: Sixty SPF-grade C57BL / 6 male mice, aged 6 to 8 weeks and weighing 18 to 22 g, were used. The experiments began after one week of adaptive feeding. The control group of mice drank purified water, while the other groups drank a 2.5% dextran sodium sulfate solution to induce UC in mice.

[0179] Drug administration: The mice in the control and model groups were gavaged with distilled water, while the mice in the sulfasalazine, TRYP, and compound 21 groups were gavaged with corresponding doses of drugs at a volume of 10 mL / kg for 12 consecutive days.

[0180] Disease Activity Index Score: From the first day of animal modeling, the weight, stool texture, and blood in the stool of the mice were recorded according to the DAI score table to perform the disease activity index (DAI) score. Figures 3 to 6 shown.

[0181] The effect of compound 21 obtained in Example 12 on the colon histopathological score of UC mice is shown in the figure below. Figure 3 As shown. Figure 3 It can be seen that the therapeutic effect of compound 21 is better than that of the positive control drug sulfasalazine.

[0182] The effect of compound 21 obtained in Example 12 on the body weight of mice with ulcerative colitis is as follows: Figure 4 As shown. Figure 4 It can be seen that compared with the model group and the sulfasalazine group, the average body weight of mice treated with compound 21 was the heaviest.

[0183] The effect of compound 21 obtained in Example 12 on the colon length of mice with ulcerative colitis is as follows: Figure 5 and Figure 6 As shown. Figure 5 and Figure 6 It can be seen that compared with the model group and the sulfasalazine group, the colon length of mice treated with compound 21 was the longest.

[0184] Combine Figures 3 to 6 It can be seen that compound 21 has a therapeutic effect on ulcerative colitis and has the potential to be developed into a drug.

[0185] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as within the scope of protection of the present invention.

Claims

1. A tryptanthrin compound, characterized in that The structural formula of the tryptanthrin compound is as follows: Said R1, R2 and R3 are independently hydrogen, halogen, methoxy, methyl, nitroso or trifluoromethoxy.

2. A tryptanthrin compound according to claim 1, characterized in that The structural formula of the tryptanthrin compounds includes:

3. The method for preparing a tryptanthrin compound according to claim 1 or 2, characterized in that: The steps include: Tryptanthrin compounds are obtained by reacting an indole-2,3-dione derivative, 2H-thieno[3,2-D][1,3]oxazine-2,4(1H)-dione, triethylamine and toluene.

4. The method for preparing a tryptanthrin compound according to claim 3, wherein: The structural formula of the indole-2,3-dione derivative is: Said R1, R2 and R3 are independently hydrogen, halogen, methoxy, methyl, nitroso or trifluoromethoxy.

5. The method for preparing a tryptanthrin compound according to claim 4, wherein: The usage ratio of the indole-2,3-dione derivative, 2H-thieno[3,2-D][1,3]oxazine-2,4(1H)-dione, triethylamine and toluene is 0.8-1.2 mmol: 0.8-1.2 mmol: 0.8-1.2 mL: 8-12 mL.

6. The method for preparing a tryptanthrin compound according to claim 4 or 5, characterized in that: The reaction time is 4 to 8 hours, and the reaction temperature is 80 to 100°C.

7. Use of a tryptanthrin compound according to claim 1 or 2 in the preparation of anti-inflammatory drugs.

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