Pyrroloquinoline fused heteroazasugar derivatives, synthetic methods and uses thereof

Abstract

The application provides a kind of pyrroloquinoxaline fused heteroazasugar derivative and its synthesis method and application.The structural formula of the pyrroloquinoxaline fused heteroazasugar derivative is shown as formula (I)-(V), and the preparation method is efficient, simple and easy to scale production.It has been verified by test that the compound of the application has good anti-tumor cell proliferation activity, and can be applied to the preparation of anti-tumor drug preparation, which will provide more drug selection for clinical treatment of tumor-related diseases.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of medicine, and particularly relates to a kind of pyrroloquinoxaline fused heteroazasugar derivatives and a synthesis method and application thereof. BACKGROUND

[0002] In the field of medicine and pharmacy today, the research and development of antitumor drugs has always been the focus of attention. As one of the diseases that seriously threaten human health, tumor constantly prompts researchers to work hard to find more effective and more targeted treatment methods and drugs.

[0003] Traditional antitumor drugs have achieved some success to some extent, but often have various limitations, such as greater side effects, easy drug resistance, etc. Therefore, it is of great significance to develop new antitumor compounds.

[0004] Azasugars, i.e. imino sugars, are widely present in plants and microorganisms in nature. Due to their unique structure and potential biological activity, such compounds and their derivatives have shown great potential for the treatment of diabetes, viral infection and immune system disorders, and have attracted widespread attention in recent years. However, existing azasugar derivatives still have some deficiencies in practical application, such as complex preparation process, low efficiency, or unsatisfactory antitumor activity, etc.

[0005] Therefore, it is of great significance to develop novel and unique nitrogen-containing fused heterocyclic compounds and study their antitumor cell proliferation activity for the research and development of antitumor drugs. SUMMARY

[0006] The purpose of the present application is to provide a kind of pyrroloquinoxaline fused heteroazasugar derivatives and a synthesis method thereof, and to provide the application of the derivatives in the preparation of antitumor drug preparations, so as to provide more drug options for clinical antitumor.

[0007] The purpose of the present application is achieved by the following technical solutions:

[0008] A kind of pyrroloquinoxaline fused heteroazasugar derivatives, the structural formula is shown as formula (I)~formula (V).

[0009]

[0010] The present application also provides a synthesis method of the above-mentioned pyrroloquinoxaline fused heteroazasugar derivatives, wherein the synthesis of the compound shown in formula (I) is as follows:

[0011]

[0012] The synthesis of the compound shown in 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 product is sequentially extracted, washed, dried, filtered, and separated by 200-300 mesh silica gel column chromatography to obtain the compounds represented by formula (I) to (V).

[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 pyrroloquinoxalinofused azasugar derivative in the preparation of anti-tumor pharmaceutical 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 pyrroloquinoxalinofused 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 a pyrroloquinoxalino-fused azasugar derivative represented by formula (V) in the preparation of pharmaceutical preparations for treating HCT116, SUN398 and SGC7901 tumors.

[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 the active ingredient, it can be combined with water, sucrose, sorbitol, fructose and other components to prepare oral liquid preparations; and 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 prescribe dosages based on individual patient differences.

[0035] The present invention provides a class of pyrroloquinoxaline-fused azasugar derivatives with an efficient and simple preparation method. The compounds and their isomers can be obtained in pure form by column chromatography, making them easy to manufacture on a large scale. Experimental verification has shown that the compounds of the present invention exhibit strong inhibitory activity against HCT116, with inhibition rates exceeding 50%. In addition to significantly inhibiting HCT116 (with an inhibition rate of 98%), the compound represented by formula (V) also exhibits very strong inhibitory activity against SUN398 and SGC7901. This invention will provide more drug options for the clinical treatment of tumor-related diseases. DETAILED DESCRIPTION

[0036] The present invention will be further described below with reference to the following examples. The following examples are only for illustration and are not intended to 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 well known in the art, the reagents used in the examples are all analytically pure or chemically pure, and are commercially available or prepared by methods well known to those skilled in the art, and the following examples all achieve the purpose of the present application.

[0038] Example 1

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

[0040] The chemical reaction flow is as follows:

[0041]

[0042] The specific method is:

[0043] D-ribose (344.4 mg, 1 mmol, commercially available or prepared by taking D-ribose as raw material, referring 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) were weighed into a 50 mL reaction bottle, 2 mL of acetonitrile was added to stir and dissolve, and the reaction was stirred at room temperature under nitrogen protection for 48 hours. The reaction was monitored by TLC. The reaction solution was diluted with water, extracted with ethyl acetate (10 mL x 3), and the organic phase was collected and washed with saturated brine. The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated by rotary evaporation. The residue was separated by silica gel column chromatography (V 石油醚 :V 乙酸乙酯 = 3:1) to obtain a light yellow solid, which was compound (I).

[0044] Compound (I): light yellow solid, yield 24%; (c 0.1, CH3OH); 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 N2O3([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] Propylidene-protected and toluenesulfonated (Ts-) D-ribose (344.4 mg, 1 mmol, commercially available or prepared from D-ribose according to the literature method of 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) were weighed into a 50 mL reaction flask and dissolved with 2 mL of methanol. 1 equivalent of acetic acid was added under nitrogen protection, and the mixture was stirred in an oil bath at 50°C-60°C for 1 hour. The reaction was monitored by TLC for completion. The reaction solution was diluted with water and extracted with ethyl acetate (10 mL x 3). The organic phase was collected and washed with saturated sodium bicarbonate solution and brine. The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was evaporated to remove the solvent, and the residue was separated 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,CH3OH); 1 H NMR(400MHz, Methanol-d4)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=1 1.3, 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-d4)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 N2O3([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] Propylidene-protected and mesylated (Ms) L-rhamnose (282.1 mg, 1 mmol, commercially available or prepared from L-rhamnose according to the literature 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) were weighed into a 50 mL reaction flask and dissolved with 2 mL of methanol. 0.2 equivalents of trifluoroacetic acid were added under nitrogen protection, and the mixture was stirred in an oil bath at 50°C-60°C for 1 hour. The reaction was monitored by TLC for completion. The reaction solution was diluted with water and extracted with ethyl acetate (10 mL x 3). The organic phase was collected and washed with saturated brine. The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was evaporated to remove the solvent, and the residue was 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,CH3OH); 1 H NMR (400MHz, Methanol-d4) 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); 13C NMR (101 MHz, Methanol-d4) δ 135.1, 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 N2O3 ([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 scheme is as follows:

[0062]

[0063] The specific method is as follows:

[0064] Take propylidene-protected and mesylated (Ms) L-rhamnose (282.1 mg, 1 mmol, commercially available or prepared from L-rhamnose according to the literature method 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 eq), and synthesize compound (IV) according to the method of Example 3.

[0065] Compound (IV): yellow solid, yield 31%; (c 0.1, CH3OH). 1H NMR (400 MHz, Methanol-d4) δ 7.47 (d, J = 1.4 Hz, 1H), 7.39 (d, J = 1.8 Hz, 2H), 7.27 (d, J = 3.1 Hz, 1H), 6.30 (d, J = 3.1 Hz, 1H), 4.69 (d, J = 7.1 Hz, 1H), 4.11 (dd, J = 3.1, 1.6 Hz, 1H), 4.05 - 3.98 (m, 1H), 3.88 (d, J = 1.1 Hz, 3H), 3.79 (dd, J = 3.5, 1.6 Hz, 1H), 3.67 (dd, J = 7.1, 3.1 Hz, 1H), 3.02 (p, J = 6.9 Hz, 1H), 1.25 (t, J = 6.1 Hz, 6H), 1.20 (d, J = 6.9 Hz, 3H). 13 C NMR (101 MHz, Methanol-d4) δ 128.8, 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 N2O5 ([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- tetrahydridopyrrolo[1,2-a:2',1'-c]quinoxalin-1,2-diol)

[0068] The chemical reaction scheme is as follows:

[0069]

[0070] The specific method is as follows:

[0071] Propylidyl-protected and mesylated (Ms) L-rhamnose (282.1 mg, 1 mmol, commercially available or prepared from L-rhamnose according to the literature method 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 eq) were weighed out and compound (V) was synthesized according to the method of Example 3.

[0072] Compound (V): light yellow solid, yield 64%; [a] 2D 5 -37.0(c 0.1,CH3OH); 1 H NMR(400MHz, Methanol-d4)δ7.51(d,J=2.3Hz,1H),7.28–7.24(m,1H),7.21(d,J =8.6Hz,1H),7.11(dd,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-d4) 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 BrN2O3([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 was calculated according to the following formula:

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

[0078] Test results: see Table 1

[0079] Table 1 Antiproliferative inhibitory 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 all exhibited strong inhibitory activity against HCT116, with inhibition rates exceeding 50%. Compounds (I) and (III) also exhibited strong inhibitory activity against SW480. In addition to significantly inhibiting HCT116 (with an inhibition rate of 98%), compound (V) also exhibited very strong inhibitory activity against SUN398 and SGC7901.

[0082] Example 9

[0083] 5 mg of 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 methods of pyrroloquinoxalino-fused azasugar derivatives and the inhibitory activities of such compounds against the proliferation of HCT116, SW480, 7901, and SNU398 tumor cells. These examples not only illustrate the synthesis methods and anti-proliferation activities of the specific compounds described herein, but also serve to illustrate the synthesis of homologues and analogues thereof by varying the types and quantities of the raw materials. They do not constitute any limitation on the scope of the present invention.

Claims

1. A pyrroloquinoxalino-fused azasugar derivative, characterized in that: Its structural formula is shown in formula (I), formula (III) or formula (V): 。 2. The method for synthesizing the pyrroloquinoxalino-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 (III) is as follows: ； The synthesis of the compound represented by formula (V) is as follows: 。 3. The method for synthesizing the pyrroloquinoxalino-fused azasugar derivative according to claim 2, wherein: 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 pyrroloquinoxalino-fused azasugar derivative according to claim 2, wherein: In the synthesis of the compound represented by formula (III), the molar ratio of compound 1b to compound 2 was 1:1.2; in the synthesis of the compound represented by formula (V), the molar ratio of compound 1b to compound 5 was 1:1.

2.

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

2.

6. The method for synthesizing the pyrroloquinoxalino-fused azasugar derivative according to claim 2, wherein: After the chemical reaction is complete, the product is sequentially extracted, washed, dried, filtered, and separated by 200-300 mesh silica gel column chromatography to obtain the compound represented by formula (I), formula (III) or formula (V).

7. The method for synthesizing the pyrroloquinoxalino-fused azasugar derivative according to claim 6, wherein: 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 compound represented by formula (III) or formula (V) using a 200-300 mesh silica gel column is V 二氯甲烷 :V 甲醇 =(10~15):

1.

8. Use of the pyrroloquinoxalino-fused azasugar derivative according to claim 1 in the preparation of an anti-tumor pharmaceutical preparation, characterized in that: When the pyrroloquinoxaline-fused azasugar derivative is a compound represented by formula (I) or formula (III), the tumor is human colon cancer; when the pyrroloquinoxaline-fused azasugar derivative is a compound represented by formula (V), the tumor is human colon cancer, human gastric adenocarcinoma or human liver cancer.

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