Synthesis method and application of 6-site alpha-hydroxyalkylated purine nucleoside compound

Abstract

The invention discloses a synthesis method and application of a 6-site alpha-hydroxyalkylated purine nucleoside compound, and belongs to the field of organic chemistry. A 9-site substituted purine compound 1 and an alcohol compound 2 are used as raw materials, and a constant-current electrolytic reaction is performed in a non-separation electrolytic tank, NHPI as a hydrogen reagent and an acetonitrile solvent to obtain a 6-site alpha-hydroxyalkylated purine nucleoside compound 3. The method is mild in reaction condition and environment-friendly, the 6-site alpha-hydroxyalkylated purine nucleoside product can be obtained with high yield, a simple and effective synthesis way is provided for the 6-site alpha-hydroxyalkylated purine nucleoside product, and meanwhile, the compound has better half inhibitory concentration on HL60 cells.

Description

Technical Field

[0001] This invention relates to a method for synthesizing and applying 6-α-hydroxyalkylated purine nucleoside compounds, belonging to the field of organic chemistry. Background Technology

[0002] C6-hydroxyalkyl purines and purine nucleosides have attracted considerable attention due to their potential applications in organic and medicinal chemistry. For example, C6-hydroxymethyl-9-(β-D-ribofuranosyl)purine nucleoside, initially isolated from *Amanita muscaria*, exhibits unique biological activities, including antifungal, cytotoxic, and antiviral (vesicular stomatitis virus) activities, as well as inhibition of adenosine deaminase. Furthermore, this compound has been used as a key intermediate in the synthesis of C6-(F- / O- / N- / S-substituted methyl) and 6-carboxymethyl purine nucleosides with cytotoxic and antiviral activities.

[0003] Despite their attractive properties, efficient methods for synthesizing C6-hydroxyalkylated purine derivatives remain limited. Traditional synthetic methods for introducing hydroxymethyl groups at the C6 position of purine nucleosides include: 1) radical photoaddition of unsubstituted purines with methanol; 2) nucleophilic addition of C6-magnesiated or lithiated purines to formaldehyde; and 3) palladium-catalyzed cross-coupling of C6-halopurines with organozinc or organotin reagents. However, these methods face numerous challenges, such as low atom economy, demanding reaction conditions, narrow substrate applicability, poor functional group tolerance, and lengthy synthetic routes, which limit their scope of synthetic applications.

[0004] Therefore, it is still very necessary to develop a green, environmentally friendly, economical and efficient method for preparing 6-α-hydroxyalkylated purine nucleoside compounds and to study their preliminary biological activities. Summary of the Invention

[0005] To overcome the above-mentioned technical defects, this invention discloses a method for preparing 6-α-hydroxyalkylated purine nucleoside compounds. The method uses 9-substituted purine compound 1 and alcohol compound 2 as raw materials, and obtains 6-α-hydroxyalkylated purine nucleoside compounds under electroanodic oxidation conditions and catalysis by a hydrogen-extracting reagent. The method has mild reaction conditions, is environmentally friendly, has high yield, good purity, and is suitable for industrial production.

[0006] The 6-α-hydroxyalkylated purine nucleoside compound of the present invention has the following general structural formula:

[0007]

[0008] Wherein: R1 is selected from benzyl, 4-acrylbenzyl, C1-C5 alkyl, allyl, propargyl, acetone, acetonyl, methoxyethoxymethyl, cyclobutylmethyl, ethoxycarbonylmethyl, phenyl, R2 is selected from hydrogen, C1-C6 alkyl or cycloalkyl, R3 is selected from hydrogen, C1-C6 alkyl, or cycloalkyl.

[0009] This invention provides a method for synthesizing a 6-α-hydroxyalkylated purine nucleoside compound, comprising the following steps: using a 9-substituted purine compound 1 and an alcohol compound 2 as raw materials, a constant current electrolysis reaction is carried out in a non-separating electrolytic cell in the presence of a hydrogen-extracting reagent, an additive, and an organic solvent to obtain a 6-α-hydroxyalkylated purine nucleoside compound 3; the reaction equation is as follows:

[0010]

[0011] Wherein, the substituents R1, R2 and R3 are the same as those described above.

[0012] Furthermore, in the above technical solution, the hydrogen-sequestering reagent is selected from TMSN3, DABCO, NHPI, Cl4NHPI, NHS or HOBt; preferably from NHPI.

[0013] Furthermore, in the above technical solution, the organic solvent is selected from acetonitrile, 1,2-dichloroethane, acetone or dimethyl sulfoxide; preferably acetonitrile.

[0014] Furthermore, in the above technical solution, the additive is selected from p-toluenesulfonic acid, acetic acid, formic acid, or trifluoroacetic acid. Trifluoroacetic acid is preferred.

[0015] Furthermore, in the above technical solution, the electrolyte in the non-separated electrolytic cell is selected from tetra-n-butylhexafluorophosphonic acid, tetra-tetrabutyltetrafluoroborate ammonium, tetraethyltetrafluoroborate ammonium, tetraethylhexafluorophosphonic acid, and tetraethylperchlorate ammonium; preferably tetraethyltetrafluoroborate ammonium.

[0016] Furthermore, in the above technical solution, the constant current refers to the power supply output current being constant, and the constant current output current is 3-10mA; preferably 5mA.

[0017] Furthermore, in the above technical solution, the molar ratio of the 9-substituted purine compound 1 to the alcohol compound 2 is 1:5.0-8.5.

[0018] Furthermore, in the above technical solution, the electrode anode material is selected from platinum sheet, RVC (reticulated glassy carbon), and GF (carbon felt); preferably GF (carbon felt).

[0019] Furthermore, in the above technical solution, the electrode cathode material is selected from platinum sheet, graphite carbon rod, and GF (carbon felt); preferably from platinum sheet.

[0020] Furthermore, in the above technical solution, the constant current electrolysis reaction temperature is selected from room temperature.

[0021] Furthermore, in the above technical solution, the progress of the constant current electrolysis reaction can be monitored using conventional monitoring methods in the art (such as TLC, HPLC or NMR), and the reaction endpoint is generally defined as when the compound disappears or when the disubstituted product begins to form.

[0022] Furthermore, in the above technical solution, if a crude compound is obtained after the reaction, it can be separated and purified by conventional methods such as preparative HPLC, preparative TLC, or recrystallization.

[0023] This invention also provides the application of 6-α-hydroxyalkylated purine nucleoside compounds in the preparation of drugs with anti-HL60 cell activity.

[0024] Furthermore, in the above technical solution, the 6-hydroxyalkylated purine nucleoside compound is selected from the following specific structures:

[0025]

[0026] Beneficial effects of the invention

[0027] The method for preparing 6-α-hydroxyalkylated purine nucleoside compounds of the present invention is green, environmentally friendly, economical and efficient, with high yield and good purity. It avoids the use of traditional toxic oxidants and yields a wide variety of 6-α-hydroxyalkylated purine nucleoside compounds that exhibit good HL60 activity, showing potential for large-scale application. Detailed Implementation

[0028] The present invention is further illustrated below by way of embodiments, but the invention is not limited to the scope of the embodiments described herein. Experimental methods in the following embodiments that do not specify specific conditions were performed according to conventional methods and conditions, or as selected according to the product instructions.

[0029] Example 1

[0030] Condition optimization experiment

[0031] In a non-separated electrolyzer, substrate 1a (42.0 mg, 0.20 mmol) and alcohol 2 (0.1 mL), NHPI (13.3 mg, 40 mol%), Et4NBF4 (86.8 mg, 0.4 mmol), and TFA (31 μL, 0.4 mmol) were dissolved in a solvent. Electrolysis was carried out at room temperature using a constant current until the substrate was completely consumed (as measured by TLC or...). 1 (H NMR analysis monitoring). After the reaction was complete, the solvent was removed under reduced pressure. The resulting residue was purified by silica gel column chromatography (PE / EA: 5 / 1-1 / 1) to obtain a white solid 3a; results under different reaction conditions are as follows:

[0032]

[0033]

[0034]

[0035] The reaction conditions are as follows: substrate 1a (42.0 mg, 0.2 mmol), substrate 2a (0.1 mL), NHPI (13.3 mg, 40 mol%), TFA (31 μL, 0.4 mmol), and acetonitrile solvent system; carbon felt anode (1.0 × 1.5 × 0.3 cm). 3 ) and platinum cathode (1.0×1.5×0.03cm) 3 In an undivided electrolytic cell, a constant current of 5mA is applied for 5 hours under nitrogen atmosphere at room temperature. b 1,3,5-Trimethoxybenzene was used as an internal standard to determine the NMR yield; c. the yield in parentheses is the separation yield; d. no reaction; e. the reaction was carried out for 13 hours with electrolysis.

[0036] The optimal conditions were finally determined: carbon felt anode (1.0*1.5*0.3cm) 3 ) and platinum cathode (1.0*1.5*0.03cm) 3 Compounds 1a (0.20 mmol), 2a (0.1 mL), NHPI (40 mol%), TFA (0.40 mmol), MeCN (3 mL), and 5 mA were reacted at room temperature under nitrogen atmosphere for 5 h.

[0037] Example 2

[0038]

[0039] In a non-separated electrolytic cell, substrate 1a (42.0 mg), 2a (0.1 mL), NHPI (13.3 mg, 40 mol%), Et4NBF4 (86.8 mg, 0.4 mmol), and TFA (31 μL, 0.4 mmol) were dissolved in acetonitrile. Electrolysis was carried out at a constant current of 5.0 mA for 5 h under nitrogen atmosphere at room temperature. After the reaction was completed, the solvent was removed under reduced pressure, and the product was purified by silica gel column chromatography (PE / EA: 5 / 1-1 / 1) to obtain a white solid 3a (49.4 mg, yield 92%). 1 H NMR (600MHz, CDCl3): δ8.94(s,1H),8.04(s,1H),7.40-7.32(m,5H),5.46(s,2H),5.44(s,1H),1.78(s,6H). 13C NMR (100MHz, CDCl3): δ165.8,151.8,151.6,143.6,135.1,129.8,129.4,128.9,128.1,72.4,47.6,29.8.

[0040] Example 3

[0041]

[0042] In a non-separated electrolytic cell, substrate 1p (75.7 mg, 0.2 mmol), 2m (5.0 eq), NHPI (13.3 mg, 40 mol%), Et4NBF4 (86.8 mg, 0.4 mmol), and TFA (31 μL, 0.4 mmol) were dissolved in acetonitrile. Electrolysis was carried out at a constant current of 5.0 mA for 7.5 h under nitrogen atmosphere at room temperature. After the reaction was completed, the solvent was removed under reduced pressure, and the product was purified by silica gel column chromatography (PE / EA: 5 / 1-1 / 1) to obtain 4 L (86.5 mg, yield 72%) of a colorless oil. 1 H NMR (400MHz, CDCl3): δ9.08(s,1H),8.41(s,1H),8.00-7.91(m,2H),6.91-6.84(m,2H),6.27(d,J=5.2Hz,1H),5.95(t,J=5.2Hz,1H),5 .64(t,J=5.2Hz,1H),4.57-4.29(m,6H),3.83(s,3H),3.52(t,J=7.2Hz,2H),2.35-2.24(m,2H),2.15(s,3H),2.12(s,3H),2.07(s,3H). 13 C NMR (100MHz, CDCl3): δ120.0,170.4,169.7,169.5,166.4,163.4,153.8,152.4,149.3,146.0 ,131.7,122.7,113.6,86.8,80.6,73.2,70.6,64.0,63.0,55.5,36.8,23.2,20.9,20.7,20.5.

[0043] Example 4

[0044]

[0045] Referring to the reaction conditions in Example 3, starting from 1p (75.7 mg, 0.2 mmol) and 2p (5.0 eq), the reaction was carried out for 6.5 h to obtain a colorless oil 4o (105.3 mg, yield 87%). 1H NMR (600MHz, CDCl3): δ8.87 (d, J = 12.0Hz, 1H), 8.20 (s, 1H), 7.51-7.46 (m, 1H), 7.4 1-7.35(m,1H),7.31-7.25(m,2H),6.22-6.16(m,1H),5.95-5.76(m,1H),5.72-5.62 (m,1H),5.62-5.54(m,1H),4.51-4.32(m,5H),3.04-2.90(m,1H),2.55-2.44(m,1H ),2.16(d,J=1.8Hz,3H),2.12(d,J=3.0Hz,3H),2.10(s,3H),1.79(d,J=2.4Hz,3H). 13 C NMR (150MHz, CDCl3): δ170.33,170.27,169.6,169.43,169.41,165.3,165.1,164.8 ,164.6,151.5,151.3,151.1,150.9,142.4,141.9,139.11,139.09,130.7,130.53,1 30.47,130.4,128.6,128.54,128.51,128.4,86.7,86.3,80.4,73.5,73.1,72.9,72.5,70.5,70.4,62.99,62.97,61.5,61.2,40.14,40.07,29.7,29.6,20.8,20.6,20.5.

[0046] Example 5

[0047]

[0048] Referring to the reaction conditions in Example 3, starting from 1p (75.7 mg, 0.2 mmol) and 2q (5.0 eq), the reaction was carried out for 5.5 h to obtain a colorless oily substance 4p (89.5 mg, yield 88%). 1H NMR (400MHz, CDCl3): δ8.89 (s, 1H), 8.20 (d, J = 2.4Hz, 1H), 6.24 (d, J = 5.2Hz, 1H), 6.01-5.89(m,1H),5.75-5.61(m,1H),5.49(s,1H),4.47-4.28(m,3H),3.99-3.84 (m,2H),3.60(dd,J=15.6,6.0Hz,1H),3.04(d,J=15.6Hz,1H),2.12(d,J=1.6Hz,3 H),2.08(s,3H),2.06(d,J=1.6Hz,3H),1.70(d,J=2.0Hz,3H),1.09-1.02(m,3H). 13 C NMR (100MHz, CDCl3): δ170.9,170.3,169.62,169.61,169.40,169.40,164.4,169.3,151.51,151.47,151.2,151.1,142.3,130.59 ,130.58,86.6,86.5,80.42,80.40,73.2,73.1,73.0,72.9,70.63,70.55,63.10,63.05,60.3,45.9,29.2,20.8,20.6,20.5,14.0.

[0049] Example 6

[0050]

[0051] In a non-separated electrolytic cell, substrate 1p (75.7 mg, 0.2 mmol), 2u (0.1 mL), NHPI (33.3 mg, 0.2 mmol), Et4NBF4 (86.8 mg, 0.4 mmol), and TFA (31 μL, 0.4 mmol) were dissolved in acetonitrile. Electrolysis was carried out at a constant current of 5.0 mA for 9 h under nitrogen atmosphere at room temperature. After the reaction was completed, the solvent was removed under reduced pressure, and the product was purified by silica gel column chromatography (PE / EA: 5 / 1-1 / 1) to obtain a colorless oily substance 4t (36.8 mg, yield 45%). 1 H NMR (600MHz, CDCl3): δ8.92(s,1H),8.22(s,1H),6.23(d,J=9.6Hz,1H),5.95(t,J=5.4Hz,1H), 5.65(t,J=5.4Hz,1H),5.18(s,2H),4.47-4.33(m,3H),2.13(s,3H),2.09(s,3H),2.06(s,3H). 13C NMR (150MHz, CDCl3): δ170.4,169.7,169.5,159.8,152.3,150.5,143.0,131.3,86.7,80.5,73.2,70.6,63.1,61.3,20.8,20.6,20.5.

[0052] Example 7

[0053]

[0054] In a non-separated electrolytic cell, substrate 1p (75.7 mg, 0.2 mmol), 2v (5.0 eq), NHPI (33.3 mg, 0.2 mmol), Et4NBF4 (86.8 mg, 0.4 mmol), and TFA (31 μL, 0.4 mmol) were dissolved in acetonitrile. Electrolysis was carried out at a constant current of 5.0 mA for 6.5 h under nitrogen atmosphere at room temperature. After the reaction was completed, the solvent was removed under reduced pressure, and the product was purified by silica gel column chromatography (PE / EA: 5 / 1-1 / 1) to obtain a colorless oily substance 4u (65.2 mg, yield 48%). 1 H NMR (400MHz, CDCl3): δ8.98-8.88(m,1H),8.24-8.17(m,1H),7.53-7.46(m,2H),7. 46-7.30(m,4H),7.15-7.04(m,2H),6.28-6.19(m,1H),6.02-5.90(m,1H),5.70-5.6 3(m,1H),5.36(s,1H),4.56-4.29(m,6H),3.71-3.57(m,1H),2.59-2.46(m,1H),2.1 6(s,3H),2.10(d,J=1.2Hz,3H),2.09-2.05(m,3H),1.72(s,1H),1.52-1.44(m,3H). 13 C NMR (100MHz, CDCl3): δ174.01,173.97,170.4,169.7,169.5,161.7,159.8(d,CF, 1 J C-F =246.6Hz),152.1,150.9,143.0,142.9,141.9(d,CF, 3 J C-F =8.0Hz), 135.7, 130.9(d,CF, 4 J C-F =3.0Hz), 129.1(d,CF, 4 J C-F=4.0Hz),128.5,127.7,123.7,115.4(d,CF, 2 J C-F =23.4Hz),86.78,86.73,86.7,80.58,80.55,73.2,70.7(d,CF, 4 J C-F =3.0Hz),67.84,67.81,63.1,61.6,45.1,36.1,20.9,20.7,20.5,18.50,18.48,18.4. 19 F NMR (376MHz, CDCl3): δ-117.8.

[0055] Example 8

[0056]

[0057] Referring to the reaction conditions in Example 7, starting from 1p (75.7 mg, 0.2 mmol) and 2w (5.0 eq), the reaction was carried out for 6.5 h to obtain a colorless oily substance 4v (43.7 mg, yield 32%). 1 H NMR (400MHz, CDCl3): δ9.10(s,1H),8.40(s,1H),7.20(d,J=8.0Hz,2H),7.08(d,J=8.0Hz,2H),6.28(d, J=5.2Hz,1H),5.97(t,J=5.6Hz,1H),5.66(t,J=5.2Hz,1H),4.51-4.37(m,3H),4.07(t,J=6.8Hz,2H),3. 68(q,J=7.2Hz,1H),3.29(t,J=7.2Hz,1H),2.42(d,J=7.2Hz,2H),2.16(s,3H),2.13(s,3H),2.08(s,3H) ,1.85-1.74(m,3H),1.69-1.60(m,4H),1.48(d,J=6.8Hz,3H),1.43-1.37(m,2H),0.87(d,J=6.8Hz,6H). 13 C NMR (100MHz, CDCl3): δ175.0,170.4,169.7,169.5,152.5,149.8,145.9,140.6,138.0,129.4,127.3,86 .8,80.7,73.2,70.7,64.7,63.1,45.3,45.2,40.1,30.3,28.5,25.6,23.4,22.5,20.9,20.7,20.5,18.6.

[0058] Example 9

[0059]

[0060] Referring to the reaction conditions in Example 7, starting from 1p (75.7 mg, 0.2 mmol) and 2x (5.0 eq), the reaction was carried out for 11 h to obtain a colorless oily substance 4v (69.4 mg, yield 51%). 1 H NMR (400MHz, CDCl3): δ8.90(s,1H),8.22(s,1H),6.25(d,J=5.2Hz,1H),5.96(t,J=5.6Hz,1H),5.67(t,J=5.2Hz,1H),5.24(s,1H),4.49- 4.35(m,3H),2.65-2.54(m,1H),2.53-2.38(m,2H),2.15(s,3H),2.12(s,3H),2.08(s,3H),2.04-1.24(m,21H),1.08(s,3H),0.88(s,3H). 13 C NMR (100MHz, CDCl3): δ170.4,169.7,169.5,166.1,151.5,151.3,142.0,130.9,86.6,80.5,73.9,73.2,70.7,63.2,5 4.2,51.7,48.0,41.0,40.1,36.1,36.0,35.3,34.1,33.5,31.8,31.0,28.3,21.9,20.9,20.7,20.5,20.4,14.0,11.8.

[0061] Example 10

[0062] Using the same reaction conditions as in Example 2 above, but changing only the reaction substrate, a variety of substituted compounds were obtained. The reaction results are as follows:

[0063]

[0064]

[0065] Example 11

[0066] The anticancer activity of the compounds was assessed using CCK8 assays via antiproliferative activity studies. First, cells were seeded at a density of 5000 cells per well into 96-well plates containing 100 μL of culture medium and incubated overnight at 37°C and 5% CO2. The next day, 100 μL of culture medium diluted with different concentrations of the test compound was added to each well. The cells were then incubated at 37°C and 5% CO2 for 72 hours. Next, 10 μL of CCK8 was added to each well, and the 96-well plates were incubated at 37°C for 2 hours. Absorbance at 450 nm was measured using a Perkin Elmer microplate reader, and IC50 values ​​were calculated using GraphPadPrism 6.0 software. All experiments were performed in triplicate. HL60 cells were selected as the study subject, and 5-fluorouracil (5-FU) was used as a positive control. The anticancer activity results of some compounds are as follows:

[0067]

[0068]

[0069] The activity results indicate that the 6-α-hydroxyalkylated purine nucleoside compound 3 provided by this invention can inhibit the proliferation activity of HL60 cells, with 3j, 4a, and 4t exhibiting significant inhibitory effects. These activity results suggest that this type of compound has medicinal value in preventing / treating / inhibiting the progression of cancer.

[0070] The above embodiments describe the basic principles, main features, and advantages of the present invention. Those skilled in the art should understand that the present invention is not limited to the above embodiments. The embodiments and descriptions in the specification are merely illustrative of the principles of the present invention. Various changes and modifications can be made to the present invention without departing from its principles, and all such changes and modifications fall within the scope of protection of the present invention.

Claims

A 1,6-α-hydroxyalkylated purine nucleoside compound, characterized in that, Its general structural formula is as follows: Wherein: R1 is selected from benzyl, 4-acrylbenzyl, C1-C5 alkyl, allyl, propargyl, acetone, acetonyl, methoxyethoxymethyl, cyclobutylmethyl, ethoxycarbonylmethyl, phenyl, R2 is selected from hydrogen, C1-C6 alkyl or cycloalkyl, R3 is selected from hydrogen, C1-C6 alkyl, or cycloalkyl.

2. The method for synthesizing the 6-α-hydroxyalkylated purine nucleoside compound as described in claim 1, characterized in that, The process includes the following steps: using 9-substituted purine compound 1 and alcohol compound 2 as raw materials, a constant current electrolysis reaction is carried out in a non-separating electrolytic cell in the presence of a hydrogen-extracting reagent, additives, and organic solvents to obtain 6-α-hydroxyalkylated purine nucleoside compound 3; the reaction equation is shown below: Wherein, the substituents R1, R2 and R3 are consistent with those described in claim 1.

3. The method for synthesizing the 6-α-hydroxyalkylated purine nucleoside compound according to claim 2, characterized in that: The hydrogen-extracting reagent is selected from NHPI, the organic solvent is selected from acetonitrile, and the additive is selected from trifluoroacetic acid.

4. The method for synthesizing the 6-α-hydroxyalkylated purine nucleoside compound according to claim 2, characterized in that: In the non-separated electrolytic cell, the electrolyte is selected from tetrabutyltetrafluoroboronic acid.

5. The method for synthesizing the 6-α-hydroxyalkylated purine nucleoside compound according to claim 2, characterized in that: The molar ratio of the 9-substituted purine compound 1 to the alcohol compound 2 is 1:5.0-8.

5.

6. The method for synthesizing the 6-α-hydroxyalkylated purine nucleoside compound according to claim 2, characterized in that: The constant current refers to the power supply output current being constant, with a constant current output current of 5mA.

7. The method for synthesizing the 6-α-hydroxyalkylated purine nucleoside compound according to claim 2, characterized in that: The anode material of the electrode is selected from platinum sheet, mesh glassy carbon, and carbon felt; the cathode material of the electrode is selected from platinum sheet, graphite carbon rod, and carbon felt.

8. The method for synthesizing the 6-α-hydroxyalkylated purine nucleoside compound according to claim 2, characterized in that: The electrolysis reaction is carried out at room temperature.

9. The use of the 6-α-hydroxyalkylated purine nucleoside compound as described in claim 1 in the preparation of an anti-HL60 cell active drug.

10. The use of the 6-α-hydroxyalkylated purine nucleoside compound according to claim 9 in the preparation of an anti-HL60 cell active drug, characterized in that: The 6-α-hydroxyalkylated purine nucleoside compound is selected from...

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