Method for synthesizing benzoxycarbonyl basic group

Abstract

The invention discloses a method for synthesizing a benzoxycarbonyl basic group. The method comprises the following steps of dissolving a nucleobase into a solvent; adding protecting reagent N-(benzyloxycarbonyloxy)succinimide; controlling the reaction temperature to 0-65 DEG C and reacting for 4-24 hours at normal pressure. The invention has the advantages of mild reaction condition, short time, simple preparation, high yield, environmental protection, no corrosion on equipment in the preparation process, simple process, and the like.

Description

technical field

[0001] The invention relates to a method for synthesizing a benzyloxycarbonyl base by one-step reaction using benzyloxycarbonyl succinimide as a raw material. Background technique

[0002] Since Nielsen et al. first designed and synthesized the nucleic acid analog peptide nucleic acid (PNA) in 1991, due to its excellent hybridization characteristics, it has been widely used in the research of biochips, gene drugs, molecular biology tools and other fields in the past ten years. In order to expand the application of PNA, research on new synthesis methods of PNA and molecular modification of PNA is still going on. So far, the price of PNA monomer is still very expensive, which limits its wide application.

[0003] When synthesizing peptide nucleic acid monomers, the active amino groups of cytosine and adenine in natural nucleobases must be protected. The most widely used protecting group is benzyloxycarbonyl chloride. The conditions for synthesizing benzyloxyc...

Examples

Embodiment 1

[0012] Synthesis of Benzyloxycarbonylcytosine

[0013]

[0014] Dissolve 10g (91mmol) of cytosine in 350ml of dry N,N-dimethylformamide, add 25g (100mmol) of benzyloxycarbonylsuccinimide in batches after 10 minutes, react at 65°C for 24 hours, evaporate under reduced pressure As a solvent, add 150ml of water, adjust the pH to 1~2 with 4M hydrochloric acid, filter with suction, wash with water twice to obtain a white light yellow solid, add 300ml of ethanol, reflux at 80°C for 15 minutes, filter, wash twice with ethanol, and wash twice with ether. 20.16 g of white solid was obtained, and the yield was 90.65%. m.p>250℃; FT-IR: V C-H : 2964V C＝O : 1745, 1693, V C-O : 1203; ESI-MS: [M+H] + =246.25; 1 HNMR (DMSO-d6): δ7.78(d, 1H), 7.36(m, 5H), 6.91(d, 1H), 5.16(s, 2H, CH 2 ).

Embodiment 2

[0016] Synthesis of benzyloxycarbonyl adenine:

[0017]

[0018] Adenine 5g (37mmol) was dissolved in ice-cold 150ml N,N-dimethylformamide, under vigorous stirring, 10g (40mmol) of benzyloxycarbonylsuccinimide was added in batches, after stirring for 4 hours, pour into 300ml In ice water, adjust the pH to 7 with 1M hydrochloric acid, filter, wash the precipitate with water, recrystallize from a mixture of methanol and chloroform, refrigerate at -4°C for 12 hours, and filter to obtain 5.22 g of a white product, with a yield of 40%. m.p=214℃; FT-IR: V C-H : 3297, 3112, V C-H : 2923V C＝O : 1754, V C-O : 1220; [M+H] + =270.13; 1 HNMR (DMSO-d6): δ: 8.11 (d, 2H), 7.275 (s, 2H), 5.052 (d, 2H), 4.15 (q, 2H), 1.199 (t, 3H).

technical field

[0001] The invention relates to a method for synthesizing a benzyloxycarbonyl base by one-step reaction using benzyloxycarbonyl succinimide as a raw material. Background technique

[0002] Since Nielsen et al. first designed and synthesized the nucleic acid analog peptide nucleic acid (PNA) in 1991, due to its excellent hybridization characteristics, it has been widely used in the research of biochips, gene drugs, molecular biology tools and other fields in the past ten years. In order to expand the application of PNA, research on new synthesis methods of PNA and molecular modification of PNA is still going on. So far, the price of PNA monomer is still very expensive, which limits its wide application.

[0003] When synthesizing peptide nucleic acid monomers, the active amino groups of cytosine and adenine in natural nucleobases must be protected. The most widely used protecting group is benzyloxycarbonyl chloride. The conditions for synthesizing benzyloxyc...

Examples

Embodiment 1

[0012] Synthesis of Benzyloxycarbonylcytosine

[0013]

[0014] Dissolve 10g (91mmol) of cytosine in 350ml of dry N,N-dimethylformamide, add 25g (100mmol) of benzyloxycarbonylsuccinimide in batches after 10 minutes, react at 65°C for 24 hours, evaporate under reduced pressure As a solvent, add 150ml of water, adjust the pH to 1~2 with 4M hydrochloric acid, filter with suction, wash with water twice to obtain a white light yellow solid, add 300ml of ethanol, reflux at 80°C for 15 minutes, filter, wash twice with ethanol, and wash twice with ether. 20.16 g of white solid was obtained, and the yield was 90.65%. m.p>250℃; FT-IR: V C-H : 2964V C＝O : 1745, 1693, V C-O : 1203; ESI-MS: [M+H] + =246.25; 1 HNMR (DMSO-d6): δ7.78(d, 1H), 7.36(m, 5H), 6.91(d, 1H), 5.16(s, 2H, CH 2 ).

Embodiment 2

[0016] Synthesis of benzyloxycarbonyl adenine:

[0017]

[0018] Adenine 5g (37mmol) was dissolved in ice-cold 150ml N,N-dimethylformamide, under vigorous stirring, 10g (40mmol) of benzyloxycarbonylsuccinimide was added in batches, after stirring for 4 hours, pour into 300ml In ice water, adjust the pH to 7 with 1M hydrochloric acid, filter, wash the precipitate with water, recrystallize from a mixture of methanol and chloroform, refrigerate at -4°C for 12 hours, and filter to obtain 5.22 g of a white product, with a yield of 40%. m.p=214℃; FT-IR: V C-H : 3297, 3112, V C-H : 2923V C＝O : 1754, V C-O : 1220; [M+H] + =270.13; 1 HNMR (DMSO-d6): δ: 8.11 (d, 2H), 7.275 (s, 2H), 5.052 (d, 2H), 4.15 (q, 2H), 1.199 (t, 3H).

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