A pyrano[2,3-a]phenazine derivative, and a preparation method and use thereof

Abstract

This invention belongs to the field of medicinal chemistry and pharmaceutical technology, and discloses a pyrano[2,3-a]phenazine derivative, its preparation method, and its application in the preparation of anti-glioma drugs. The structural formula of the pyrano[2,3-a]phenazine derivative is as follows: R1 is H, halogen, or -N(C2H5)2; R2 is H or halogen; R3 is H, halogen, or -CH3; R4 is H or halogen; R5 is H, halogen, or -OCH3; R6 is H, halogen, -CH3, -C2H5, or -OCH3; R7 is H or -OCH3; X is -CN or -COOC2H5. This invention also discloses the preparation method of this pyrano[2,3-a]phenazine derivative and its application in the preparation of anti-glioma drugs. This type of compound has a strong inhibitory effect on thioredoxin reductase (TrxR) and significant inhibitory activity against human glioma cells (U87), showing great application potential in the preparation of anti-glioma drugs.

Description

Technical Field

[0001] This invention relates to the fields of medicinal chemistry and pharmaceutical technology, specifically to a class of novel pyrano[2,3-a]phenazine derivatives with anti-glioma activity, as well as methods for preparing these compounds and their pharmaceutical uses. Background Technology

[0002] Cancer remains one of the most threatening diseases to humankind. By 2040, an estimated 28.4 million new cancer cases are projected globally. In 2020, an estimated 19.3 million new cancer cases were diagnosed worldwide (excluding non-melanoma skin cancer), and approximately 10 million people died from cancer. Glioblastoma, a highly malignant brain tumor, is genetically unstable and highly invasive, exhibiting resistance to chemotherapy. Its incidence is 2-3 per 100,000 adults, with a median survival of only 12-15 months, making it one of the most difficult primary tumors of the central nervous system to treat. Current clinical treatments for cancer include surgical resection, radiotherapy, and chemotherapy. Chemotherapy, which kills primary and metastatic cancer cells with chemotherapeutic drugs, is highly efficient compared to other treatments and has become a major method for treating cancer. However, as treatment continues, tumors often develop resistance to chemotherapeutic compounds. Therefore, overcoming tumor resistance and finding chemotherapeutic drugs targeting different targets is urgently needed.

[0003] The thioredoxin (Trx / TrxR) system is one of the most important systems regulating redox balance in cells. Studies have found that TrxR is highly expressed in various primary tumors, including liver cancer, lung cancer, and glioma, and is associated with tumor growth and proliferation, drug resistance, and poor patient prognosis. Therefore, TrxR has become an effective target for cancer treatment. Compared with healthy cells, cancer cells are characterized by elevated levels of reactive oxygen species (ROS), caused by their uncontrolled proliferation and intense metabolism. Under normal physiological conditions, cells maintain redox homeostasis by regulating the balance between intracellular oxidants and antioxidants, resulting in low levels of ROS. However, excessive ROS can also be cytotoxic, and cancer cells may be more sensitive to drugs that increase ROS production. Therefore, targeting TrxR with exogenous substances to induce ROS increase and selectively kill cancer cells has become an effective therapeutic strategy. Summary of the Invention

[0004] To address the aforementioned technical problems in the prior art, this invention provides a series of pyrano[2,3-a]phenazine derivatives and their preparation methods, and provides the application of these compounds in the preparation of anti-glioma drugs.

[0005] The technical solution of this invention is as follows:

[0006] The first object of this invention is to provide a pyrano[2,3-a]phenazine derivative having the structural formula shown in Formula I:

[0007] in,

[0008] R1 is H, halogen, or -N(C2H5)2;

[0009] R2 is H or a halogen;

[0010] R3 can be H, halogen, or -CH3;

[0011] R4 is H or a halogen;

[0012] R5 is H, halogen, or -OCH3;

[0013] R6 can be H, halogen, -CH3, -C2H5, or -OCH3;

[0014] R7 is H or -OCH3;

[0015] X is either -CN or -COOC2H5.

[0016] Furthermore, the halogen is fluorine, chlorine, or bromine.

[0017] Furthermore, the pyrano[2,3-a]phenazine derivative is selected from compounds shown in the following structural formulas:

[0018]

[0019]

[0020] Furthermore, the pyrano[2,3-a]phenazine derivative is selected from compounds shown in the following structural formulas:

[0021]

[0022]

[0023] A second object of the present invention is to provide the use of the pyrano[2,3-a]phenazine derivative of Formula I in the preparation of anti-glioma drugs.

[0024] A third object of the present invention is to provide a method for preparing the pyrano[2,3-a]phenazine derivative of formula I, comprising the following steps:

[0025] S1. Aniline, o-toluidine, 3-bromo-2-methylaniline, 2,4-dichloroaniline, and N,N-diethyl-p-phenylenediamine react with 5-chloro-2-nitrobenzene ether in the presence of potassium tert-butoxide to give compounds a, b, c, d, and e, respectively. The general structural formulas of compounds a, b, c, d, and e are shown in Formula II.

[0026] The reaction process is as follows:

[0027]

[0028] S2. The products a, b, c, d, and e of S1 are reacted with N,O-bis(trimethylsilyl)acetamide (BSA) to obtain compounds with the general structural formula shown in Formula III. Then, under the action of boron tribromide, compounds with the general structural formula shown in Formula IV are obtained.

[0029]

[0030] The reaction process is as follows:

[0031]

[0032] S3: The product of S2 is reacted with a nitrile derivative and an aromatic aldehyde. The product is purified to obtain the pyrano[2,3-a]phenazine derivative of formula I.

[0033] In steps (1) and (2), R1 is H, a halogen, or -N(C2H5)2; R2 is H or a halogen; and R3 is H, a halogen, or -CH3.

[0034] In a particular embodiment:

[0035] In compounds a corresponding to formulas II to IV, R1 is H; R2 is H; and R3 is H.

[0036] In compounds b corresponding to formulas II to IV, R1 is H; R2 is H; and R3 is -CH3.

[0037] In compounds c corresponding to formulas II to IV, R1 is H; R2 is Br; and R3 is -CH3.

[0038] In compounds d corresponding to formulas II to IV, R1 is Cl; R2 is H; and R3 is Cl.

[0039] In compounds e corresponding to formulas II to IV, R1 is -N(CH2CH3)2; R2 is H; and R3 is H.

[0040] Furthermore, the purification is performed by recrystallization.

[0041] Compared with the prior art, the present invention has the following beneficial effects:

[0042] (1) Provide a series of novel pyrano[2,3-a]phenazine derivatives and apply them to anti-glioma drugs.

[0043] (2) The method provided is simple and efficient, and can quickly construct a variety of derivatives with both pyran and phenazine active skeletons.

[0044] (3) Experiments have shown that the novel pyrano[2,3-a]phenazine derivative involved in this invention has a significant inhibitory effect on the U87 glioma cell line and has broad application potential in the preparation of anticancer drugs. Detailed Implementation

[0045] The present invention will be further explained below with reference to the embodiments, but the embodiments do not limit the present invention in any way.

[0046] Example 1: Synthesis of compounds a, b, c, d, and e

[0047] The stir bar and flask were dried at 100°C for 1 hour beforehand. Potassium tert-butoxide (4 eq.) was added to the flask under nitrogen protection, followed by the addition of an appropriate amount of N,N-dimethylformamide solution. The flask was then placed in a cryogenic reactor and stirred for 20 minutes at -60°C. After stirring, aniline compounds (aniline, o-toluidine, 3-bromo-2-methylaniline, 2,4-dichloroaniline, or N,N-diethyl-p-phenylenediamine) dissolved in N,N-dimethylformamide (1.1 eq.) were slowly added dropwise. After the addition was complete, the mixture was stirred for 10 minutes, followed by the addition of a 5-chloro-2-nitrosoanisole (1 eq.) solution in N,N-dimethylformamide. Stirring continued at -60°C for 3-4 hours. After the reaction was complete, the resulting mixture was extracted with saturated ammonium chloride aqueous solution and ethyl acetate. The mixture was washed three times with saturated brine, and the organic layers were combined and dried with anhydrous sodium sulfate. The mixture was filtered and concentrated under vacuum to obtain the crude nitroso compound target product, which was used directly in the next step without purification.

[0048] The target products of the crude nitroso compounds are compound a obtained from aniline, compound b obtained from o-toluidine, compound c obtained from 3-bromo-2-methylaniline, compound d obtained from 2,4-dichloroaniline, and compound e obtained from N,N-diethyl-p-phenylenediamine.

[0049] Example 2: Synthesis of compounds a1, b1, c1, d1, and e1

[0050] Crude nitroso compound (1 eq) was dissolved in an appropriate amount of N,N-dimethylformamide under nitrogen protection, and then N,O-bis(trimethylsilyl)acetamide (BSA, 5 eq) was added. The resulting mixture was stirred at 50 °C for 16 h until the reaction was complete. An appropriate amount of water was added to the reaction mixture, and stirring was continued at room temperature for 10 min. After solid precipitation, the precipitate was filtered with cold ethyl acetate and washed, and the filter cake was collected. The filtrate was extracted with saturated brine and ethyl acetate, washed three times with brine, and the organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The concentrated product and filter cake were combined and purified by silica gel column chromatography to obtain the target product with the general formula compound III.

[0051] The target products of the general structural formula compound III are compound a1 obtained from compound a, compound b1 obtained from compound b, compound c1 obtained from compound c, compound d1 obtained from compound d, and compound e1 obtained from compound e.

[0052] Example 3: Synthesis of compounds a2, b2, c2, d2, and e2

[0053] The round-bottom flask and stir bar were dried in an oven at 100°C for 1 hour beforehand. Compound (1 eq) with the general formula (III) was dissolved in anhydrous dichloromethane under nitrogen protection. The solution was placed in a -78°C cryogenic reactor and stirred for 20 minutes. 1M boron tribromide solution (BBr3, 6 eq) was added dropwise. The reaction mixture was stirred at -78°C for 1 hour and then allowed to stand overnight at room temperature. After overnight reaction, the mixture was heated to reflux at 40°C until the reaction was complete. The reaction was quenched with an appropriate amount of saturated brine and extracted with dichloromethane. The organic layer was dried with anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain the phenazine core compound.

[0054] The phenazine nucleus compounds are compound a2 obtained from compound a1, compound b2 obtained from compound b1, compound c2 obtained from compound c1, compound d2 obtained from compound d1, and compound e2 obtained from compound e1.

[0055] Example 4: Synthesis of compound H1

[0056] A one-pot, multi-component reaction was employed. Compound a2, 3,4-dichlorobenzaldehyde (1 eq), malononitrile (1 eq), and triethylenediamine (DABCO, 1 eq) were added to anhydrous ethanol and refluxed for 12 h. After the reaction was complete, the precipitate was cooled, filtered, and recrystallized from hot ethanol to obtain compound H1.

[0057] Yield: 35%. 1H NMR(300MHz,DMSO-d6)δ8.3(dt,J=6.6,3.3Hz,2H),8.2(s,1H),8.1(dd,J=6.8,3.4H z,2H),7.6(d,J=8.3Hz,1H),7.5–7.5(m,3H),7.2(dd,J=8.4,2.2Hz,1H),5.1(s,1H). 13 C NMR (126MHz, DMSO-d6) δ159.9,145.6,145.5,143.9,142.8,142.5,135.3,133.9,132.8,1 32.4,131.8,131.6,130.3,129.9,129.8,129.7,128.4,125.2,119.9,119.4,57.4,40.3.

[0058]

[0059] Example 5: Synthesis of compound H2

[0060] The method is the same as in Example 4, except that compound b2 is used instead of compound a2 to obtain compound H2.

[0061] Yield: 59%. 1 H NMR (300MHz, DMSO-d6) δ8.2 (s, 1H), 8.1 (dd, J = 8.6, 1.7Hz, 1H), 8.0–7.9 (m, 2H), 7. 6(s,1H),7.5(d,J=5.6Hz,3H),7.2(dd,J=8.4,2.2Hz,1H),5.1(s,1H),2.8(s,3H). 13 C NMR(126MHz,Chloroform-d)δ159.0,144.4,144.1,143.2,143.0,141.6,138.3,134.8,133.2,1 32.7,131.8,131.7,131.1,130.9,129.6,127.5,127.2,126.2,119.5,118.5,61.5,39.8,17.6.

[0062]

[0063] Example 6: Synthesis of compound H3

[0064] The method is the same as in Example 4, except that compound c2 is used instead of compound a2 to obtain compound H3.

[0065] Yield: 41%. 1H NMR(300MHz,Chloroform-d)δ8.2(s,1H),8.0(s,2H),7.4(d,J=8.3Hz,1H),7.3( d,J=2.2Hz,1H),7.2(dd,J=8.3,2.2Hz,1H),5.2(s,2H),5.1(s,1H),3.0(s,3H). 13 CNMR(126MHz,Chloroform-d)δ158.8,144.5,143.8,142.8,142.1,141.8,137.9,136.2,135.6, 133.2,132.6,131.9,130.9,129.6,128.0,128.0,127.2,126.2,119.9,118.4,61.6,39.8,17.4.

[0066]

[0067] Example 7: Synthesis of compound H4

[0068] The method is the same as in Example 4, except that compound d2 is used instead of compound a2 to obtain compound H4.

[0069] Yield: 20%. 1 H NMR (300MHz, DMSO-d6) δ8.4(d,J=2.2Hz,1H),8.2–8.2(m,2H),7.6(d,J=8.3Hz,1H),7.5–7.5(m,3H),7.2(dd,J=8.4,2.2Hz,1H),5.1(s,1H). 13 C NMR(126MHz,Chloroform-d)δ158.7,144.4,143.1,142.6,142.0,139.7,136.9,136.6,134. 6,133.9,133.3,132.5,132.1,131.0,129.6,127.3,127.2,126.2,121.2,118.2,61.6,39.8.

[0070]

[0071] Example 8: Synthesis of compound H5

[0072] The method is the same as in Example 4, except that compound e2 is used instead of compound a2 to obtain compound H5.

[0073] Yield: 50%. 1H NMR (300MHz, DMSO-d6) δ8.0(d,J=9.7Hz,1H),8.0(s,1H),7.8(dd,J=9.7,2.7Hz,1H),7.6(d,J=8.3Hz,1H),7.5(d,J=2.1Hz ,1H),7.4(s,2H),7.1(dd,J=8.3,2.2Hz,1H),6.9(d,J=2.4Hz,1H),5.0(s,1H),3.6(d,J=7.1Hz,4H),1.3(t,J=6.8Hz,6H). 13 C NMR (126MHz, DMSO-d6) δ160.2,149.7,145.8,145.1,144.4,141.3,139.5,134.0,131.7,131.6,130.6 ,130.4,130.2,129.8,127.7,125.3,124.8,120.2,119.1,100.6,57.3,45.0,45.0,40.3,13.0,13.0.

[0074]

[0075] Example 9: Synthesis of compound H6

[0076] The method is the same as in Example 4, except that compound e2 is used instead of compound a2, and o-chlorobenzaldehyde is used instead of 3,4-dichlorobenzaldehyde, to obtain compound H6.

[0077] Yield: 53.2%. 1 H NMR(300MHz,DMSO-d6)δ8.0(d,J=9.7Hz,1H),7.9(s,1H),7.8(dd,J=9.8,2.8Hz,1H),7.5–7.4(m,1H),7.3 –7.2(m,4H),7.0–7.0(m,1H),6.9(d,J=2.7Hz,1H),5.5(s,1H),3.6(q,J=7.1Hz,4H),1.3(t,J=6.9Hz,6H). 13 C NMR (151MHz, DMSO-d6) δ160.3,149.7,145.5,145.0,141.8,139.9,139.3,133.8,132.4,130.6,130. 5,130.0,129.2,128.6,125.3,124.9,119.8,119.7,116.6,100.6,56.5,45.0,45.0,13.0,13.0,6.4.

[0078]

[0079] Example 10: Synthesis of compound H7

[0080] The method is the same as in Example 4, except that compound e2 is used instead of compound a2, and p-chlorobenzaldehyde is used instead of 3,4-dichlorobenzaldehyde, to obtain compound H7.

[0081] Yield: 26.4%. 1 H NMR(300MHz,Chloroform-d)δ8.0(d,J=9.8Hz,1H),8.0(s,1H),7.6–7.6(m,1H),7.3(s,1H),7 .3(s,1H),7.2–7.1(m,3H),5.1(d,J=20.2Hz,3H),3.6(q,J=7.1Hz,4H),1.4(t,J=6.9Hz,6H). 13 C NMR(151MHz,DMSO-d6)δ160.1,149.7,145.5,144.5,143.7,139.9,139.3,133.9,132.0,130.6,130.5 ,129.7,129.7,129.2,129.2,125.2,125.0,120.2,119.9,100.5,57.7,45.0,45.0,13.0,13.0,13.0.

[0082]

[0083] Example 11: Synthesis of compound H8

[0084] The method is the same as in Example 4, except that compound e2 is used instead of compound a2, and 2,3-dichlorobenzaldehyde is used instead of 3,4-dichlorobenzaldehyde, to obtain compound H8.

[0085] Yield: 72%. 1 H NMR (300MHz, DMSO-d6) δ8.0(d,J=9.7Hz,1H),8.0(s,1H),7.8(dd,J=9.8,2.8Hz,1H),7.5(dd,J=8.0,1.5Hz,1H),7.4(s, 2H),7.3(t,J=7.9Hz,1H),7.0–7.0(m,1H),6.9(d,J=2.7Hz,1H),5.6(s,1H),3.6(d,J=7.3Hz,4H),1.2(d,J=6.8Hz,6H). 13CNMR(151MHz,DMSO-d6)δ160.3,149.7,145.5,145.0,140.0,139.4,133.8,132.5,130.6,130.6,130. 5,130.3,129.7,129.4,125.3,125.0,119.7,119.3,100.5,100.5,56.0,45.0,45.0,13.0,13.0,13.0.

[0086]

[0087] Example 12: Synthesis of compound H9

[0088] The method is the same as in Example 4, except that compound e2 is used instead of compound a2, and benzaldehyde is used instead of 3,4-dichlorobenzaldehyde to obtain compound H9.

[0089] Yield: 34.2%. 1 H NMR(300MHz,DMSO-d6)δ8.0(d,J=9.7Hz,1H),7.9(s,1H),7.8(dd,J=9.7,2.7Hz,1H),7.4–7.2(m, 5H),7.2–7.1(m,2H),6.9(d,J=2.7Hz,1H),4.9(s,1H),3.6(t,J=7.3Hz,4H),1.2(d,J=6.8Hz,6H). 13 C NMR(151MHz,DMSO-d6)δ160.1,149.6,145.5,144.7,144.5,139.8,139.3,133.9,130.7,130.6,129.2 ,129.2,127.8,127.8,127.5,125.2,124.9,120.5,120.4,100.6,58.2,45.0,45.0,13.0,13.0,13.0.

[0090]

[0091] Example 13: Synthesis of compound H10

[0092] The method is the same as in Example 4, except that compound e2 is used instead of compound a2, and 2-bromobenzaldehyde is used instead of 3,4-dichlorobenzaldehyde, to obtain compound H10.

[0093] Yield: 60%. 1H NMR(300MHz,Chloroform-d)δ8.0(s,1H),8.0(s,1H),7.6(d,J=8.2Hz,2H),7.2(s,1H),7.1(s ,1H),7.0(t,J=8.5Hz,2H),5.7(s,1H),5.1(s,2H),3.6(d,J=7.6Hz,4H),1.4(d,J=3.4Hz,6H). 13 C NMR(151MHz,DMSO-d6)δ172.5,160.2,149.7,145.5,144.9,143.6,139.9,139.3,133.9,133.3,130.6 ,130.5,129.4,129.2,128.2,125.3,124.9,124.9,123.2,100.6,56.5,45.0,45.0,21.5,13.0,13.0.

[0094]

[0095] Example 14: Synthesis of compound H11

[0096] The method is the same as in Example 4, except that compound e2 is used instead of compound a2, and 4-bromobenzaldehyde is used instead of 3,4-dichlorobenzaldehyde, to obtain compound H11.

[0097] Yield: 31.1%. 1 H NMR(300MHz,DMSO-d6)δ8.0(d,J=9.7Hz,1H),7.9(s,1H),7.8(dd,J=9.8,2.8Hz,1H),7.5–7.5(m,2H),7 .3(s,2H),7.2–7.1(m,2H),6.9(d,J=2.6Hz,1H),5.0(s,1H),3.6(q,J=7.0Hz,4H),1.2(d,J=7.2Hz,6H). 13 C NMR (126MHz, DMSO-d6) δ160.1,149.7,145.5,144.5,144.1,141.2,138.2,133.9,132.2,132.2,130.6 ,130.5,130.1,130.1,125.3,125.0,120.6,119.8,108.1,100.6,57.7,45.0,45.0,13.0,13.0,13.0.

[0098]

[0099] Example 15: Synthesis of compound H12

[0100] The method is the same as in Example 4, except that compound e2 is used instead of compound a2, and 3-bromo-4,5-dimethoxybenzaldehyde is used instead of 3,4-dichlorobenzaldehyde, to obtain compound H12.

[0101] Yield: 44%. 1 H NMR (300MHz, DMSO-d6) δ8.0(d,J=9.7Hz,1H),7.9(s,1H),7.8(dd,J=9.8,2.8Hz,1H),7.3(s,2H),6.9(dd,J=6.2, 2.3Hz,2H),6.8(d,J=1.9Hz,1H),5.0(s,1H),3.8(s,3H),3.7(s,3H),3.6(t,J=7.8Hz,4H),1.2(t,J=7.0Hz,6H). 13 C NMR(126MHz,Chloroform-d)δ166.0,158.7,154.4,150.2,149.9,149.3,147.1,144.7,144.1,138.7,135.3,1 29.9,129.4,127.7,125.0,124.4,122.1,117.8,117.0,105.4,65.3,65.3,62.5,61.3,61.3,49.7,17.7,17.7.

[0102]

[0103] Example 16: Synthesis of compound H13

[0104] The method is the same as in Example 4, except that compound e2 is used instead of compound a2, and 4-methylbenzaldehyde is used instead of 3,4-dichlorobenzaldehyde, to obtain compound H13.

[0105] Yield: 26%. 1 H NMR (300MHz, DMSO-d6) δ8.0(d,J=9.7Hz,1H),7.9(s,1H),7.8(dd,J=9.8,2.8Hz,1H),7.2(s,2H),7.1(d,J=8.0Hz, 2H),7.0(d,J=8.1Hz,2H),6.9(d,J=2.7Hz,1H),4.9(s,1H),3.6(q,J=7.0Hz,4H),2.2(s,3H),1.3(t,J=6.9Hz,6H). 13C NMR(126MHz,Chloroform-d)δ164.9,164.9,154.4,150.2,149.1,146.5,144.6,144.0,141.4,138.6,135.5 ,134.5,134.5,132.4,132.4,132.4,125.4,107.7,105.3,105.3,63.2,49.7,49.7,25.8,17.7,17.7,17.7.

[0106]

[0107] Example 17: Synthesis of compound H14

[0108] The method is the same as in Example 4, except that compound e2 is used instead of compound a2, and 4-ethylbenzaldehyde is used instead of 3,4-dichlorobenzaldehyde, to obtain compound H14.

[0109] Yield: 37%. 1 H NMR (300MHz, DMSO-d6) δ8.0(d,J=9.7Hz,1H),7.9(s,1H),7.8(dd,J=9.8,2.8Hz,1H),7.2(s,2H),7.1(d,J=8.2Hz,2H),7.1(d ,J=8.2Hz,2H),6.9(d,J=2.7Hz,1H),4.9(s,1H),3.6(q,J=7.0Hz,4H),2.6(s,2H),1.3(t,J=6.9Hz,6H),1.1(t,J=7.6Hz,3H).

[0110]

[0111] Example 18: Synthesis of compound H15

[0112] The method is the same as in Example 4, except that compound e2 is used instead of compound a2, 3-fluoro-4-methoxybenzaldehyde is used instead of 3,4-dichlorobenzaldehyde, and ethyl cyanoacetate malononitrile is used instead to obtain compound H15.

[0113] Yield: 19%. 1H NMR(300MHz, DMSO-d6)δ8.0(d,J=9.7Hz,1H),7.9(s,1H),7.9–7.8(m,3H),7.4–7.2(m,1H),7.1–7.0(m,2H),6.9( d,J=3.0Hz,1H),5.2(s,1H),4.1(ddd,J=10.7,6.9,3.5Hz,2H),3.8(s,3H),3.6(d,J=7.1Hz,4H),1.3–1.2(m,9H). 13 C NMR (126MHz, Chloroform-d) δ172.9,165.6,157.2,154.3,152.3,151.0,150.7,150.3,149.0,144.5,143.9,143. 7,143.4,138.7,135.5,129.2,129.2,127.7,118.9,105.4,82.4,64.3,61.1,49.7,36.4,35.1,19.6,17.8,17.8.

[0114]

[0115] Example 19: Synthesis of compound H16

[0116] The method is the same as in Example 4, except that compound e2 is used instead of compound a2, 2,4-dichlorobenzaldehyde is used instead of 3,4-dichlorobenzaldehyde, and ethyl cyanoacetate malononitrile is used instead to obtain compound H16.

[0117] Yield: 53%. 1 H NMR(300MHz,DMSO-d6)δ8.0(d,J=9.6Hz,3H),7.9(s,1H),7.8(dd,J=9.8,2.8Hz,1H),7.5(d,J=1.9Hz,1H) ,7.4–7.3(m,2H),6.9(d,J=2.6Hz,1H),5.5(s,1H),4.1–4.1(m,2H),3.6(t,J=7.3Hz,4H),1.3–1.2(m,9H).

[0118]

[0119] Example 20: Inhibitory effect of the pyrano[2,3-a]phenazine derivative described in this patent on glioma cell growth.

[0120] Using the reported TrxR inhibitor CPUL1 as a positive control, the MTT assay was used to detect the in vitro antiproliferative activity of U87 (human glioma cells). Cells were first seeded in 96-well plates and cultured for 24 hours. Then, pyrano[2,3-a]phenazine derivatives H1-H16 prepared in Examples 4-19 were added to the cells at concentration gradients of 40 μM, 20 μM, 10 μM, 5 μM, and 2.5 μM, respectively. After 48 hours of incubation, MTT solution was added to each well, and the cells were incubated at 37°C for 4 hours. The absorbance was then measured. The concentration of the compound that inhibited cell growth by 50% was calculated and expressed as IC50. 50 The values ​​are represented, and the results are shown in Table 1.

[0121] Table 1. Inhibitory effects of compounds on the growth of the central route cell line (IC50). 50 / μM)

[0122]

[0123] The results showed that the compound described in this patent has a strong inhibitory effect on the U87 cell line in vitro. Therefore, the pyrano[2,3-a]phenazine derivative described in this invention can be used to prepare anti-glioma drugs.

[0124] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A pyrano[2,3-a]phenazine derivative characterized by, The pyrano[2,3-a]phenazine derivative is selected from the compounds represented by the following structural formula: H4 H5 H6 H7 H8 H10 H11 H14.

2. Use of the pyrano[2,3-a]phenazine derivative of claim 1 in the preparation of an anti-glioma drug.

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