6-position alpha-hydroxyalkylated purine nucleoside compounds, methods of synthesis and uses thereof

Abstract

The application discloses a kind of 6 position alpha-hydroxyalkylation purine nucleoside compound synthesis method and application, belong to organic chemistry field.9 position substitution purine compound 1 and aldehyde compound 2 are used as raw material, in non-separation electrolytic cell, NHPI is used as hydrogenation reagent, acetonitrile and water mixed solvent constant current electrolysis reaction, obtain 6 position alpha-hydroxyalkylation purine nucleoside compound 3.The method of the application is mild, environment-friendly, can obtain 6 position alpha-hydroxyalkylation purine nucleoside product with high yield, provides a kind of simple and effective synthesis path for it.Simultaneously, the compound has good half inhibitory concentration to HL60 cell.

Description

Technical Field

[0001] This invention relates to a 6-α-hydroxyalkylated purine nucleoside compound, its synthesis method and application, and belongs to the field of organic chemistry. Background Technology

[0002] Purine bases and nucleosides, as fundamental building blocks of RNA and DNA, have attracted widespread attention in the fields of biochemistry and medicinal chemistry. Purine pharmacophores are common motifs in many antiviral and anticancer drugs. Among numerous derivatives, C6-hydroxyalkylated purines are a particularly important subclass, renowned for their unique structural features and valuable biological activities, and possess significant potential for drug discovery.

[0003] 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, harsh 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 from aldehydes 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 aldehyde compound 2 as raw materials, and obtains 6-α-hydroxyalkylated purine nucleoside compounds under conditions of electroanodic oxidation and cathodic reduction, and catalysis by a hydrogen-extracting reagent. The method has mild reaction conditions, is environmentally friendly, has high yield and 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, phenyl, , , R2 is selected from hydrogen, C1-C18 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 aldehyde compound 2 as raw materials, a constant-current paired electrolytic reaction is carried out in a non-separated 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 and R2 are the same as those mentioned 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, dichloromethane, and N,N-dimethylformamide; 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-n-butyltetrafluoroborate ammonium, or tetraethyltetrafluoroborate ammonium; preferably tetrabutyltetrafluoroborate 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 3-10 mA; preferably 3 mA.

[0017] Furthermore, in the above technical solution, the molar ratio of the 9-substituted purine compound 1 to the alcohol compound 2 is 1:3.0.

[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 from GF (carbon felt).

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

[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 the disappearance of the compound.

[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] Beneficial effects of the invention

[0024] 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, avoiding the use of traditional toxic oxidants, and yielding a wide variety of 6-α-hydroxyalkylated purine nucleoside compounds. Detailed Implementation

[0025] 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.

[0026] Example 1

[0027] Condition optimization experiment

[0028] In a non-separated electrolyzer, substrate 1a (42.0 mg, 0.20 mmol) and aldehyde compound 2 (0.6 mmol), NHPI (16.3 mg, 50 mol%), were mixed. n Bu4NBF4 (86.8 mg, 0.4 mmol) and TFA (31 μL, 0.6 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:

[0029]

[0030]

[0031] aThe reaction conditions were as follows: substrate 1a (42.0 mg, 0.2 mmol), substrate 2a (0.6 mmol, 3 equiv.), NHPI (50 mol%), TFA (0.6 mmol, 3 equiv.), acetonitrile solvent system, carbon felt anode (1.0 × 1.0 × 0.3 cm). 3 ) and platinum cathode (1.0 × 2.0 × 0.1 cm) 3 In an undivided electrolytic cell, a constant current of 3 mA was applied for 16 hours under nitrogen atmosphere at room temperature. bc The value in parentheses is the separation yield; c No response

[0032] The optimal conditions were finally determined: carbon felt anode (1.0*1.0*0.3 cm) 3 ) and platinum cathode (1.0*2.0*0.1cm) 3 Compounds 1a (0.20 mmol), 2a (0.6 mmol), NHPI (50 mol%), TFA (0.60 mmol), MeCN (2 mL), H2O (20 μL), and 3 mA were reacted at room temperature under nitrogen for 16 h.

[0033] Example 2

[0034]

[0035] In a non-separated electrolytic cell, substrate 1a (42.0 mg), 2a (76 μL, 0.6 mmol), and NHPI (16.3 mg, 50 mol%) were added. n Bu4NBF4 (132.7 mg, 0.4 mmol) and TFA (46 μL, 0.6 mmol) were dissolved in a mixed solvent of acetonitrile and water in a ratio of 100:1. Electrolysis was carried out under nitrogen atmosphere at room temperature with a constant current of 3.0 mA for 16 h. 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 give a yellow solid 3a (59.8 mg, yield 87%). 1H NMR (400 MHz, CDCl3): δ 8.95 (s, 1H), 8.03 (s, 1H), 7.38 -7.34 (m,3H), 7.32 - 7.30 (m, 2H), 7.22 - 7.18 (m, 4H), 7.13 - 7.10 (m, 1H), 5.45 (s,2H), 5.35 (dd, J = 8.0, 3.6 Hz, 1H), 4.65 (br, 1H), 2.90 - 2.82 (m, 2H), 2.56- 2.46 (m, 1H), 2.23 - 2.16 (m, 1H). 13 C NMR (100 MHz, CDCl3): δ 161.9, 152.0,144.1, 135.0, 130.4, 129.4, 128.9, 128.7, 128.4, 128.1, 125.9, 70.5, 47.6,39.2, 31.6, 29.9.

[0036] Example 3

[0037]

[0038] In a non-separated electrolytic cell, substrate 1m (41.7 mg, 0.2 mmol), 2b (3.0 eq), and NHPI (16.3 mg, 50 mol%) were added. n Bu4NBF4 (132.7 mg, 0.4 mmol) and TFA (46 μL, 0.6 mmol) were dissolved in a mixed solvent of acetonitrile to water at a ratio of 100:1. Electrolysis was carried out under nitrogen atmosphere at room temperature with a constant current of 3.0 mA for 16 h. 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 4a (62.7 mg, yield 88%). 1H NMR (400 MHz, CDCl3): δ 8.90 (s, 1H), 8.22 (s, 1H), 7.22 - 7.18 (m,2H), 7.14 - 7.08 (m, 3H), 5.72 (s, 2H), 5.32 (dd, J = 7.6, 4.0 Hz, 1H), 4.52(br, 1H), 3.72 - 3.68 (m, 2H), 3.51 - 3.46 (m, 2H), 3.30 (s, 3H), 2.67 - 2.60(m, 2H), 2.21 - 2.12 (m, 1H), 1.94 - 1.73 (m, 3H). 13 C NMR (100 MHz, CDCl3): δ162.2, 152.2, 151.5, 144.5, 142.3, 130.2, 128.4, 128.2, 125.7, 73.2, 71.5,70.6, 69.3, 59.1, 37.2, 35.8, 27.1, 23.78, 19.6.

[0039] Example 4

[0040]

[0041] Referring to the reaction conditions in Example 3, starting from 1m (41.7 mg, 0.2 mmol) and 2v (3.0 eq), the reaction was carried out for 14 h to give a white solid 4u (49.0 mg, yield 68%). 1 H NMR (400 MHz, CDCl3): δ 8.93 (s, 1H), 8.22 (s, 1H), 5.73 (s, 2H), 5.40 - 5.28 (m, 1H), 5.26 - 5.13 (m, 1H), 4.20 (br, 1H), 3.74 - 3.69 (m, 2H), 3.53 - 3.47 (m, 2H), 3.32 (s, 3H), 2.30 - 2.09(m, 2H), 2.06 - 1.79 (m, 3H), 1.60 - 1.54 (m, 3H), 0.92 (d, J = 6.0 Hz, 3H),0.74 (d, J = 17.6 Hz, 3H). 13C NMR (100 MHz, CDCl3): δ 163.1, 162.9, 152.5,152.4, 151.6, 148.5, 148.3, 144.5, 130.6, 122.1, 121.9, 73.2, 71.6, 70.9,70.0, 69.3, 59.2, 47.3, 47.0, 46.9, 46.3, 38.4, 36.4, 35.3, 25.6, 20.0, 19.7,12.7, 12.6.

[0042] Example 5

[0043]

[0044] Referring to the reaction conditions in Example 3, starting from 1r (75.6 mg, 0.2 mmol) and 2a (3.0 eq), the reaction was carried out for 14 h to give a white solid 3r (81.0 mg, yield 81%). 1 H NMR (400 MHz, CDCl3): δ 8.92 (s, 1H),8.21 (s, 1H), 7.22 - 7.16 (m, 4H), 7.13 - 7.08 (m, 1H), 6.23 (d, J = 5.2 Hz,1H), 5.97 - 5.93 (m, 1H), 5.67 (t, J = 4.8 Hz, 1H), 5.32 (dd, J = 8.0, 4.0Hz, 1H), 4.56 (br, 1H), 4.49 - 4.42 (m, 3H), 2.90 - 2.78 (m, 2H), 2.52 - 2.43(m, 1H), 2.24 - 2.17 (m, 1H), 2.14 (s, 3H), 2.11 (s, 3H), 2.07 (s, 3H). 13 CNMR (100 MHz, CDCl3): δ 170.4, 169.7, 169.5, 162.4, 152.1, 150.8, 142.7,141.8, 131.1, 128.6, 128.5, 128.3, 125.8 ,86.7, 86.6, 80.5, 73.2, 73.1, 70.6,70.3, 63.1, 38.9, 31.5, 20.8, 20.5.

[0045] Example 6

[0046]

[0047] Referring to the reaction conditions in Example 3, starting from 1 m (41.7 mg, 0.2 mmol) and 2 u (3.0 eq), the reaction was carried out for 14 h to give 4 t (39.5 mg, yield 62%) of white solid. 1 H NMR (400 MHz, CDCl3): δ 8.93 (s, 1H), 8.19 (s, 1H), 5.85 - 5.79 (m, 2H), 5.73 (s 2H), 3.75 - 3.72 (m, 2H), 3.72 -3.67 (m, 1H), 3.53 - 3.50 (m, 2H), 3.34 (s, 3H), 2.73 - 2.61 (m, 1H), 2.42 -2.19 (m, 4H), 2.13 - 2.07 (m, 2H). 13 C NMR (100 MHz, CDCl3): δ 166.5, 153.2,151.3, 143.8, 131.9, 126.8, 126.2, 73.1, 71.7, 69.3, 59.2, 38.0, 30.0 ,29.8,27.6, 25.5.

[0048] Example 7

[0049]

[0050] Referring to the reaction conditions in Example 3, starting from 1m (41.7 mg, 0.2 mmol) and 2w (3.0 eq), the reaction was carried out for 14h to give a white solid 4v (39.5 mg, yield 62%). 1 H NMR (400 MHz, CDCl3): 8.97 (s, 1H), 8.24 (s, 1H), 5.75 (s, 2H), 5.43 - 5.40 (m, 1H), 5.12 (d, J = 10.0 Hz, 1H), 3.76 - 3.72 (m, 2H), 3.65 (br, 1H), 3.54 - 3.50 (m, 2H), 3.34 (s, 3H), 2.56 (d, J = 6.6 Hz, 1H), 2.23 - 2.15 (m, 1H), 1.81 - 1.77 (m, 2H), 1.69 - 1.62(m, 1H), 1.59 (s, 3H), 1.09 (d, J = 6.9 Hz, 3H). 13C NMR (100 MHz, CDCl3): δ162.8, 152.8, 151.7, 144.8, 132.6, 131.3, 127.8, 73.3, 73.2, 71.7, 69.4,59.2, 44.5, 30.7, 30.3, 23.5, 20.4, 15.5.

[0051] Example 8

[0052]

[0053] Referring to the reaction conditions in Example 3, starting from 1m (41.7 mg, 0.2 mmol) and 2y (3.0 eq), the reaction was carried out for 16 h to obtain a colorless oily substance 4x (47.8 mg, yield 58%). 1 H NMR (400 MHz, CDCl3): δ 8.92 (s, 1H),8.24 (s, 1H), 7.16 - 7.11 (m, 2H), 6.96 - 6.91 (m, 2H), 5.74 (d, J = 2.4 Hz,2H), 5.25 (d, J = 4.0 Hz, 1H), 4.58 (br, 1H), 3.78 - 3.68 (m, 2H), 3.53 -3.51 (m, 2H), 3.34 (s, 3H), 2.81 - 2.73 (m, 1H), 2.64 (dd, J = 13.6, 4.4 Hz,1H), 2.42 (dd, J = 13.6, 10.0 Hz, 1H), 1.25 (s, 9H), 1.07 (d, J = 6.8 Hz, 3H). 13 C NMR (100 MHz, CDCl3): δ 161.8, 152.1, 151.5, 148.4, 144.4, 137.6,130.7, 128.9, 124.9, 75.1, 73.3, 71.7, 69.4, 59.2, 41.2, 36.4, 34.4, 31.5,17.0.

[0054] Example 9

[0055]

[0056] Referring to the reaction conditions in Example 3, starting from 1m (41.7 mg, 0.2 mmol) and 2k (3.0 eq), the reaction was carried out for 16 h to obtain a colorless oily substance 4j (27.3 mg, yield 44%).1 H NMR (400 MHz, CDCl3): δ 8.93 (s, 1H), 8.23 ​​(s, 1H), 5.74 (s, 2H), 5.34 - 5.26 (m, 1H), 4.53 (d, J = 6.4 Hz, 1H), 3.74 - 3.69 (m, 2H), 3.53 - 3.50 (m, 4H), 3.32 (s, 3H), 2.21 - 2.13 (m, 1H), 1.95 - 1.85 (m, 2H), 1.73 - 1.54 (m, 3H). 13 C NMR (100 MHz, CDCl3): δ 162.1,152.3, 151.7, 144.6, 130.2, 73.3, 71.6, 70.5, 69.4, 59.2, 45.0, 36.7, 35.6,22.8.

[0057] Example 10

[0058]

[0059] Referring to the reaction conditions in Example 3, starting from 1m (41.7 mg, 0.2 mmol) and 2r (3.0 eq), the reaction was carried out for 16 h to obtain a colorless oily substance 4s (44.5 mg, yield 69%). 1 H NMR (400 MHz, CDCl3): δ 8.91 (s, 1H), 8.17 (s, 1H), 5.71 (s, 2H), 3.88 (p, J = 8.4 Hz, 1H), 3.73 - 3.69 (m, 2H), 3.53 - 3.49 (m, 2H), 3.33 (s, 3H), 2.20 - 2.13 (m, 2H), 2.09 - 2.02 (m, 2H), 1.95 - 1.91 (m, 2H), 1.81 - 1.72 (m, 3H). 13 C NMR (100 MHz, CDCl3): δ 166.8,153.2, 151.1, 143.7, 132.2, 73.1, 71.6, 69.3, 59.2, 42.9, 32.9, 26.4.

[0060] Example 11

[0061] Using the same reaction conditions as in Example 3 above, but changing only the reaction substrate, a variety of substituted compounds were obtained. The reaction results are as follows:

[0062]

[0063]

[0064]

[0065]

[0066] Example 12

[0067] The anticancer activity of the compounds was assessed using CCK8 assay, and their antiproliferative activity was evaluated through cell culture studies. First, cells were seeded at a density of 5000 cells per well into 96-well plates containing 100 μL of culture medium per well and incubated at 37°C. o Incubate overnight at 37°C and 5% CO2. The next day, add 100 μL of culture medium diluted with different concentrations of the test compound to each well. Then incubate the cells at 37°C. o Incubate at C and 5% CO2 for 72 hours. Then, add 10 μL of CCK8 to each well and place the 96-well plate at 37°C. o Incubate at 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:

[0068]

[0069] The activity results indicate that the 6-α-hydroxyalkylated purine nucleoside compounds 3 and 4 provided in this invention can inhibit the proliferation activity of HL60 cells, with 3r, 4j, and 4s exhibiting significant inhibitory effects. These activity results suggest that these compounds have 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, phenyl, , , R2 is selected from hydrogen, C1-C18 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 aldehyde 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:

3. Wherein: the substituents R1 and R2 are the same as those described in claim 1.

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

5. 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 tetrabutylammonium tetrafluoroborate.

6. 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 aldehyde compound 2 is 1:

3.

7. 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 3mA.

8. 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 foam nickel.

9. 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.

10. 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.

11. 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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