Pyrroloquinoxaline fused azasugar derivative as well as synthesis method and application thereof

Abstract

The invention provides pyrroloquinoxaline fused azasugar derivatives as well as a synthesis method and application thereof. The structural formula of the pyrroloquinoxaline fused azasugar derivative is as shown in formulas (I)-(V), and the preparation method is efficient, simple and convenient and easy for large-scale production. Tests prove that the compound has good anti-tumor cell proliferation activity, can be applied to preparation of anti-tumor medicine preparations, and can provide more medicine choices for clinical treatment of tumor-related diseases.

Description

Technical Field

[0001] The present invention belongs to the field of pharmaceutical technology, and specifically relates to a class of pyrroloquinoxaline-fused azasugar derivatives and a synthesis method and application thereof. Background Art

[0002] In today's medical and pharmaceutical fields, the research and development of anti-tumor drugs has always been a focus of attention. As one of the diseases that seriously threaten human health, tumors have continuously prompted researchers to devote themselves to finding more effective and targeted treatment methods and drugs.

[0003] Although traditional anti-tumor drugs have achieved certain results, they often have various limitations, such as large side effects and easy to develop drug resistance. Therefore, it is of great significance to develop new anti-tumor compounds.

[0004] Azasugars, also known as iminosugars, are widely found in plants and microorganisms in nature. This class of compounds and their derivatives have shown great medicinal potential in the treatment of diabetes, viral infections and immune system disorders due to their unique structure and potential biological activity, and have attracted widespread attention in recent years. However, existing azasugar derivatives still have some shortcomings in practical applications, such as complex preparation processes, low efficiency, or unsatisfactory anti-tumor activity.

[0005] Therefore, it is of great significance to develop novel and unique nitrogen-containing hybrid compounds and study their anti-tumor cell proliferation activity for the research and development of anti-tumor drugs. Summary of the invention

[0006] The purpose of the present invention is to provide a class of pyrroloquinoxaline-fused azasugar derivatives and a synthesis method thereof, and at the same time provide the use of the derivatives in the preparation of anti-tumor drug preparations, in order to provide more drug options for clinical anti-tumor.

[0007] The objective of the present invention is achieved through the following technical solutions:

[0008] A class of pyrroloquinoxaline-fused azasugar derivatives, whose structural formulas are shown in formula (I) to formula (V).

[0009]

[0010] The present invention also provides a method for synthesizing the above-mentioned pyrroloquinoxaline-fused azasugar derivative, wherein the synthesis of the compound represented by formula (I) is as follows:

[0011]

[0012] The synthesis of the compound represented by formula (II) is as follows:

[0013]

[0014] The synthesis of the compound represented by formula (III) is as follows:

[0015]

[0016] The synthesis of the compound represented by formula (IV) is as follows:

[0017]

[0018] The synthesis of the compound represented by formula (V) is as follows:

[0019]

[0020] Furthermore, in the synthesis of the compound represented by formula (I), the molar ratio of compound 1a to compound 2 is 1:1.2.

[0021] Furthermore, in the synthesis of the compound represented by formula (II), the molar ratio of compound 1a to compound 3 is 1:1.2.

[0022] Furthermore, in the synthesis of the compound represented by formula (II), the molar ratio of compound 1a to acetic acid is 1:1.

[0023] Furthermore, in the synthesis of the compounds represented by formula (III)-(V), the molar ratio of compound 1b to compound 2 or compound 1b to compound 4 or compound 1b to compound 5 is 1:1.2.

[0024] Furthermore, in the synthesis of the compounds represented by formula (III)-(V), the molar ratio of compound 1b to trifluoroacetic acid is 1:0.2.

[0025] Furthermore, after the above chemical reaction is completed, the compounds represented by formula (I)-(V) are obtained by extraction, washing, drying, suction filtration and 200-300 mesh silica gel column chromatography.

[0026] Furthermore, the mobile phase for chromatographic separation of the compound represented by formula (I) using a 200-300 mesh silica gel column is V 石油醚 :V 乙酸乙酯 =(3-5):1; The mobile phase for chromatographic separation of the compounds represented by formula (I) to (V) using a 200-300 mesh silica gel column is V 二氯甲烷 :V 甲醇 =(10~15):1.

[0027] The present invention also provides the use of the pyrroloquinoxaline-fused azasugar derivative in the preparation of anti-tumor drug preparations.

[0028] Furthermore, the tumor is a colon tumor (HCT116), a colon tumor (SW480), a liver cancer tumor (SUN398) or a gastric gland tumor (SGC7901).

[0029] Furthermore, the present invention provides the use of pyrroloquinoxaline-fused azasugar derivatives represented by formula (I) to (IV) in the preparation of anti-HCT116 tumor drug preparations.

[0030] Furthermore, the present invention provides the use of the pyrroloquinoxaline-fused azasugar derivative represented by formula (V) in the preparation of anti-HCT116, SUN398 and SGC7901 tumor drug preparations.

[0031] The compound of the present invention is uniformly mixed with a pharmacologically acceptable carrier and can be prepared into various forms of pharmaceutical preparations for tumor-related diseases according to conventional preparation methods.

[0032] If the compound of the present invention is an active ingredient, it can be combined with water, sucrose, sorbitol, fructose and other components to prepare an oral liquid preparation; it can be combined with excipients (lactose, glucose, sucrose, mannitol), disintegrants (starch), lubricants (stearic acid, talc), binders (gelatin, polyvinyl alcohol) and other components to prepare tablets or capsules.

[0033] The compound of the present invention as an active ingredient can also be prepared into an injection solution with physiological saline, glucose solution or a mixed carrier consisting of saline and glucose.

[0034] The effective dosage of the present invention for clinical use is 10-20 mg / person / day, 2-3 times a day. Doctors can also formulate dosages based on individual differences of patients.

[0035] The present invention provides a class of pyrroloquinoxaline and fused azasugar derivatives, the preparation method of which is efficient and simple, and the compound and its isomers can be obtained by column chromatography separation, which is easy to mass produce. It has been verified by experiments that the compounds of the present invention have strong inhibitory activity against HCT116, and their inhibition rates are all above 50%. In addition to being able to significantly inhibit HCT116 (inhibition rate of 98%), the compound shown in formula (V) also shows very strong inhibitory activity against SUN398 and SGC7901. The present invention will provide more drug options for the clinical treatment of tumor-related diseases. DETAILED DESCRIPTION

[0036] The present invention is further described below in conjunction with examples. The following examples are only for illustration and do not limit the scope of protection of the present invention in any way.

[0037] The processes and methods not described in detail in the following examples are conventional methods known in the art. The reagents used in the examples are all analytically pure or chemically pure and can be purchased commercially or prepared by methods well known to those of ordinary skill in the art. The following examples all achieve the purpose of the present invention.

[0038] Example 1

[0039] Synthesis of Compound (Ⅰ) ((7R,7aR,10aS,10bS)-9,9-dimethyl-7,7a,10a,10b-tetrahydro-6H-[1,3]dioxo[4',5':3,4]pyrido[1,2-a]pyrrolo[2,1-c]quinoxalin-7-ol)

[0040] The chemical reaction process is as follows:

[0041]

[0042] The specific method is:

[0043] Weigh propylidene-protected and toluenesulfonylated (Ts) D-ribose (344.4 mg, 1 mmol, commercially available or prepared with D-ribose as raw material according to the method of Alexis Kotland, et al., J. Org. Chem. 2011, 76, 10, 4094-4098) and 1-2-aminophenylpyrrole (189.6 mg, 1.2 equivalents, commercially available) in a 50 mL reaction bottle, add 2 mL of acetonitrile and stir to dissolve, stir at room temperature under nitrogen protection for 48 hours, and monitor the reaction by TLC until complete. Dilute the reaction solution with water, extract with ethyl acetate (10 mL×3), collect the organic phase, wash with saturated brine, dry the organic phase with anhydrous sodium sulfate, filter, remove the solvent from the filtrate by rotary evaporation, and separate the residue by 200-300 mesh silica gel column chromatography (V 石油醚 :V 乙酸乙酯 =3:1) to obtain a light yellow solid, namely compound (I).

[0044] Compound (Ⅰ): light yellow solid, yield 24%; (c 0.1,CH 3 OH); 1H NMR(400MHz,Chloroform-d)δ7.36–7.32(m,1H),7.22(dd,J=3.0,1.5Hz,1H),7. 10–7.04(m,1H),6.91–6.83(m,2H),6.37(q,J=3.2,2.6Hz,1H),6.26–6.22(m,1H) ,4.47(d,J=4.6Hz,1H),4.44(d,J=9.0Hz,1H),4.23(dd,J=9.0,5.1Hz,2H),3.90 (dd,J=13.2,5.0Hz,1H),3.22(dd,J=13.1,9.9Hz,1H),1.68(s,3H),1.45(s,3H); 13 C NMR (101MHz, Chloroform-d) δ126.2,125.1,118.6,114.7,114.0,112.2,110.8,110.3,105.9,77.3,75.2,63.8,55.7,47.4,28.2,26.2; MS (ESI): C 18 H 20 N 2 O 3 ([M+H] + ):313.2.

[0045] Example 2

[0046] Synthesis of Compound (II) ((1S,2R,3R,15bS)-1,3,4-15b-tetrahydro-2H-indolo[1,2-a]pyrido[2,1-c]quinoxaline-1,2,3-triol)

[0047] The chemical reaction process is as follows:

[0048]

[0049] The specific method is:

[0050] Weigh propylidene-protected and toluenesulfonylated (Ts) D-ribose (344.4 mg, 1 mmol, commercially available or prepared with D-ribose as raw material according to the literature method Alexis Kotland, et al., J. Org. Chem. 2011, 76, 10, 4094-4098) and 2-(1H-indol-1-yl)aniline (249.6 mg, 1.2 equivalents, commercially available) in a 50 mL reaction bottle, add 2 mL of methanol and stir to dissolve, add 1 equivalent of acetic acid under nitrogen protection, and stir to react in an oil bath at 50°C-60°C for 1 hour, and monitor the reaction by TLC until it is complete. Dilute the reaction solution with water, extract with ethyl acetate (10 mL×3), collect the organic phase, wash with saturated sodium bicarbonate solution and brine, dry the organic phase with anhydrous sodium sulfate, filter, remove the solvent from the filtrate by rotary evaporation, and separate the residue by 200-300 mesh silica gel column chromatography (V 二氯甲烷 :V 甲醇 =10:1) to obtain a white solid, namely compound (II).

[0051] Compound (II): white solid, yield 62%; [α] 2 D 5 -61.0(c 0.1,CH 3 OH); 1 H NMR (400 MHz, Methanol-d 4 )δ8.00(d,J=8.4Hz,1H),7.95–7.90(m,1H),7.62(d,J=7.8Hz,1H),7.28– 7.22(m,1H),7.19–7.10(m,3H),7.04(ddd,J=8.2,6.8,2.0Hz,1H),6.70(s ,1H),4.44(d,J=10.0Hz,1H),4.14(d,J=2.7Hz,1H),3.91(ddd,J=11.3,4. 7,2.6Hz,1H),3.67(ddd,J=12.1,7.1,3.8Hz,2H),3.28(d,J=12.0Hz,1H); 13 C NMR (101MHz, Methanol-d 4 )δ137.6,135.7,130.2,124.2,121.9,120.5,120.4,119.3,116.4,113.6,111.1,100.6,71.9,69.2,65.1,54.8,46.1; MS(ESI):C 19 H 18 N 2 O 3 ([M+H]+ ):323.2.

[0052] Example 3

[0053] Synthesis of Compound (III) ((1S,2R,12bS)-3-((S)-1-hydroxyethyl)-1,2,3,12b-tetrahydrodipyrrolo[1,2-a:2',1'-c]quinoxaline-1,2-diol)

[0054] The chemical reaction process is as follows:

[0055]

[0056] The specific method is:

[0057] Weigh 282.1 mg, 1 mmol of propylidene-protected and mesylated (Ms-) L-rhamnose (282.1 mg, 1 mmol, commercially available or prepared with L-rhamnose as raw material according to the literature method Alexis Kotland, et al., J. Org. Chem. 2011, 76, 10, 4094–4098) and 1-2-aminophenylpyrrole (189.6 mg, 1.2 equivalents, commercially available) in a 50 mL reaction bottle, add 2 mL of methanol and stir to dissolve, add 0.2 equivalents of trifluoroacetic acid under nitrogen protection, and stir and react in an oil bath at 50°C-60°C for 1 hour, and monitor the reaction by TLC until it is complete. Dilute the reaction solution with water, extract with ethyl acetate (10 mL×3), collect the organic phase, wash with saturated brine, dry the organic phase with anhydrous sodium sulfate, filter, and remove the solvent by rotary evaporation of the filtrate. The residue is separated by 200-300 mesh silica gel column chromatography (V 二氯甲烷 :V 甲醇 =10:1) to obtain a light yellow solid, namely compound (III).

[0058] Compound (III): light yellow solid, yield 47%; [α] 2 D 5 -36.0(c 0.1,CH 3 OH); 1 H NMR (400 MHz, Methanol-d 4)δ7.38(dd,J=7.9,1.2Hz,1H),7.28(dd,J=2.9,1.4Hz,1H),6.97(td,J=7.6,1 .2Hz, 1H), 6.91 (dd, J=8.0, 1.4Hz, 1H), 6.82 (td, J=7.6, 1.5Hz, 1H), 6.29 (t, J= 3.2Hz,1H),6.16(dd,J=3.5,1.4Hz,1H),4.50(d,J=9.3Hz,1H),4.20–4.12(m, 2H), 3.81 (d, J = 3.0Hz, 1H), 3.73 (dd, J = 9.3, 1.9Hz, 1H), 1.32 (d, J = 6.3Hz, 3H); 13 C NMR (101MHz, Methanol-d 4 )δ135.1,125.9,124.5,118.9,116.1,114.3,114.1,109.6,105.6,86.1,79.8,78.6,77.4,53.5,12.8; MS(ESI):C 16 H 18 N 2 O 3 ([M+H] + ):287.2.

[0059] Example 4

[0060] Synthesis of Compound (Ⅳ) ((1S,2R,12bS)-1,2-dihydroxy-3-((S)-1-hydroxyethyl)-12-isopropyl-1,2,3,12b-tetrahydropyrrolo[1,2-a:2',1'-c]quinoxaline-6-carboxylic acid methyl ester)

[0061] The chemical reaction process is as follows:

[0062]

[0063] The specific method is:

[0064] Propylidene-protected and mesylated (Ms) L-rhamnose (282.1 mg, 1 mmol, commercially available or prepared with L-rhamnose as raw material according to the method of Alexis Kotland, et al., J. Org. Chem. 2011, 76, 10, 4094-4098) and methyl 3-amino-4-(1H-pyrrol-1-yl)benzoate (259.3 mg, 1.2 equivalents) were weighed, and compound (IV) was synthesized according to the method of Example 3.

[0065] Compound (IV): yellow solid, yield 31%; (c 0.1,CH 3 OH); 1 H NMR (400 MHz, Methanol-d 4 )δ7.47(d,J=1.4Hz,1H),7.39(d,J=1.8Hz,2H),7.27(d,J=3.1Hz,1H),6.30(d ,J=3.1Hz,1H),4.69(d,J=7.1Hz,1H),4.11(dd,J=3.1,1.6Hz,1H),4.05–3.98( m,1H),3.88(d,J=1.1Hz,3H),3.79(dd,J=3.5,1.6Hz,1H),3.67(dd,J=7.1,3. 1Hz, 1H), 3.02 (p, J = 6.9Hz, 1H), 1.25 (t, J = 6.1Hz, 6H), 1.20 (d, J = 6.9Hz, 3H).; 13 C NMR (101MHz, Methanol-d 4 )δ128.8,128.7,125.3,120.2,119.5,116.0,114.3,113.2,108.7,87.3,79.0,78.7,77.0,51.0,50.7,24.7,23.8,23.1,12.8; MS(ESI):C 21 H 26 N 2 O 5 ([M+H] + ):387.3.

[0066] Example 5

[0067] Synthesis of Compound (V) ((1S,R,12bS)-7-bromo-3-((S)-1-hydroxyethyl)-1,2,3,12b-tetrahydrodipyrrolo[1,2-a:2',1'-c]quinoxaline-1,2-diol)

[0068] The chemical reaction process is as follows:

[0069]

[0070] The specific method is:

[0071] Propylidene-protected and mesylated (Ms) L-rhamnose (282.1 mg, 1 mmol, commercially available or prepared with L-rhamnose as raw material according to the method of Alexis Kotland, et al., J. Org. Chem. 2011, 76, 10, 4094-4098) and 5-bromo-2-(1H-pyrrol-1-yl)aniline (283.2 mg, 1.2 equivalents) were weighed, and compound (V) was synthesized according to the method of Example 3.

[0072] Compound (V): light yellow solid, yield 64%; [α] 2 D 5 -37.0(c 0.1,CH 3 OH); 1 H NMR (400 MHz, Methanol-d 4 )δ7.51(d,J=2.3Hz,1H),7.28–7.24(m,1H),7.21(d,J=8.6Hz,1H),7.11(d d,J=8.7,2.3Hz,1H),6.32(t,J=3.2Hz,1H),6.14–6.11(m,1H),4.52(dd,J =5.4,3.2Hz,1H),4.19(dd,J=7.9,4.5Hz,1H),4.12(d,J=5.3Hz,1H),3.99 (dd,J=7.9,3.1Hz,1H),3.71(dd,J=6.7,4.7Hz,1H),0.80(d,J=6.6Hz,3H); 13 C NMR (101MHz, Methanol-d 4 )δ137.0,132.8,130.1,127.6,123.8,123.3,115.1,114.3,110.7,102.9,70.5,68.8,68.0,56.3,56.2,10.5; MS(ESI):C 16 H 17 Bn 2 O 3 ([M+H] + ):365.1.

[0073] Example 8

[0074] The compounds of the present invention were tested for their anti-proliferation activities against colon tumor (HCT116), colon tumor (SW480), liver cancer tumor (SUN398), and gastric gland tumor (SGC7901) cells.

[0075] Test method:

[0076] Cell counting kit-8 (CCK-8) is a product of Sigma and is used to measure cell viability. The experiment was divided into a blank group, a control group, and a sample group, with 3 parallels in each group. Tumor cells were inoculated into a 96-well plate at a density of 3000-5000 cells / well, and the culture medium was 100 μL. The test compounds were added to the wells at different concentrations, and the plates were incubated at 37°C for 48 hours and 96 hours, respectively. After 48 hours and 96 hours of medication, 10 μL of CCK-8 reagent was added to each well of the plate in the dark. The absorbance value at 450 nm was measured by a microplate reader to evaluate cell viability. The inhibition rate of compounds (I)-(V) on tumor cells in % was calculated according to the following formulas:

[0077] 100-[(sample group OD 450nm / Blank group OD 450nm )×100].

[0078] Test results: see Table 1

[0079] Table 1 Antiproliferation and inhibition activity of compounds (I)-(V) of the present invention on different tumor cells

[0080]

[0081] As shown in Table 1, compounds (I)-(V) of the present invention have strong inhibitory activity against HCT116, and their inhibition rates are all above 50%. Compounds (I) and (III) also have strong inhibitory activity against SW480. In addition to being able to significantly inhibit HCT116 (inhibition rate of 98%), compound (V) also shows very strong inhibitory activity against SUN398 and SGC7901.

[0082] Example 9

[0083] 5 mg of the compound (V) prepared in Example 7, 60 mg of lactose, 30 mg of potato flour, 2 mg of polyvinyl alcohol, and 1 mg of magnesium stearate were prepared into oral tablets.

[0084] Examples 1-9 listed in the present invention are intended to illustrate the preparation method of pyrroloquinoxaline-fused azasugar derivatives and the inhibitory activity of such compounds on the proliferation of HCT116, SW480, 7901 and SNU398 tumor cells. The examples are not only for illustrating the synthesis method of the specific compounds described in themselves and the anti-proliferation activity of HCT116, SW480, 7901 and SNU398 tumor cells, but also for illustrating the synthesis of homologues and analogues thereof by changing the type and amount of raw materials, and do not constitute any limitation to the scope of the present invention.

Claims

1. A pyrroloquinoxaline-fused azasugar derivative, characterized in that: Its structural formula is shown in formula (I) to formula (V).

2. The method for synthesizing the pyrroloquinoxaline-fused azasugar derivative according to claim 1, characterized in that: The synthesis of the compound represented by formula (I) is as follows: The synthesis of the compound represented by formula (II) is as follows: The synthesis of the compound represented by formula (III) is as follows: The synthesis of the compound represented by formula (IV) is as follows: The synthesis of the compound represented by formula (V) is as follows:

3. The method for synthesizing the pyrroloquinoxaline-fused azasugar derivative according to claim 2, characterized in that: In the synthesis of the compound represented by formula (I), the molar ratio of compound 1a to compound 2 is 1:1.

2.

4. The method for synthesizing the pyrroloquinoxalinofused azasugar derivative according to claim 2, characterized in that: In the synthesis of the compound represented by formula (II), the molar ratio of compound 1a to compound 3 is 1:1.

2.

5. The method for synthesizing the pyrroloquinoxalinofused azasugar derivative according to claim 2, characterized in that: In the synthesis of the compound represented by formula (II), the molar ratio of compound 1a to acetic acid is 1:

1.

6. The method for synthesizing the pyrroloquinoxalino-fused azasugar derivative according to claim 2, characterized in that: In the synthesis of the compounds represented by formula (III)-(V), the molar ratio of compound 1b to compound 2 or compound 1b to compound 4 or compound 1b to compound 5 is 1:1.

2.

7. The method for synthesizing the pyrroloquinoxalino-fused azasugar derivative according to claim 2, characterized in that: In the synthesis of the compounds represented by formula (III)-(V), the molar ratio of compound 1b to trifluoroacetic acid is 1:0.

2.

8. The method for synthesizing the pyrroloquinoxalino-fused azasugar derivative according to claim 2, characterized in that: After the chemical reaction is complete, the compounds represented by formula (I) to (V) are obtained by extraction, washing, drying, suction filtration and 200-300 mesh silica gel column chromatography.

9. The method for synthesizing the pyrroloquinoxalino-fused azasugar derivative according to claim 8, characterized in that: The mobile phase for chromatographic separation of the compound represented by formula (I) using a 200-300 mesh silica gel column is V 石油醚 :V 乙酸乙酯 =(3-5):1; The mobile phase for chromatographic separation of the compounds represented by formula (I) to (V) using a 200-300 mesh silica gel column is V 二氯甲烷 :V 甲醇 =(10~15):

1.

10. Use of the pyrroloquinoxalinofused azasugar derivative according to claim 1 in the preparation of anti-tumor pharmaceutical preparations.

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