A study on the synthesis of sulfoxides by enzyme-catalyzed oxidation of sulfides

Abstract

This invention discloses an enzyme-catalyzed method for synthesizing sulfoxides by the oxidation of thioethers. Specifically, it involves the catalytic aerobic oxidation of compound A (thioether) at room temperature to prepare compound B (sulfoxide). The preparation process of this invention is simple and low-cost, avoids the use of large amounts of oxidants and metal reagents, uses air as the oxidant, and produces water as a byproduct, thus reducing waste generation. Compared with the original process, its reaction conditions are milder, the product is less likely to be over-oxidized to sulfone, and it achieves high conversion rate, high yield, and high purity.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical intermediates, specifically relating to a method for synthesizing sulfoxides by enzyme-catalyzed oxidation of thioethers. Background Technology

[0002] Compounds containing sulfoxide units exhibit excellent biological activities against HIV-1, tumors, cell proliferation, epilepsy, hypertension, inflammation, oxidation, and parasites. In addition, these compounds are also used as metal ligands. Developing new synthetic methods for sulfoxides holds promising market potential.

[0003] The structural formulas of sulfoxide compounds are as follows:

[0004]

[0005] The synthesis of sulfoxides typically involves using strong oxidants such as hydrogen peroxide or m-CPBA with metal catalysts like sodium tungstate or titanium salts at low temperatures. Due to their strong oxidizing properties, this type of reaction often results in peroxidation to form sulfones. The resulting sulfones have similar physical and chemical properties to sulfoxides and are difficult to remove, significantly limiting the application of sulfoxides in the pharmaceutical field. Therefore, developing a method to prepare sulfoxides from sulfides without undergoing excessive oxidation holds great promise. Summary of the Invention

[0006] To overcome the shortcomings of the prior art, this invention provides an enzyme-catalyzed method for the oxidation of thioethers (compound A) to sulfoxides (compound B), which can efficiently obtain sulfoxides with high purity.

[0007] To achieve the technical objective of this invention, the technical solution is as follows: Compound A (thioether), enzyme, proton donor, coenzyme, solubilizer, and buffer solution are added to a reaction flask to prepare compound B (sulfoxide).

[0008]

[0009] The enzyme is MOX-012 and GDH enzyme; the proton donor is glucose; and the coenzyme is NADP+.

[0010] The amino acid sequence of the monooxygenase MOX-012 is shown in SEQ ID NO.1:

[0011] MDKHIDVLIVGAGISGLGLAAHLSKNCPQRSFEIVERREGIGGTWDLFRYPGIRSDSDMS

[0012] TFGYNFKPWRKAKILADGASIRQYLHEVVDEFHLDRKIHFKHRVISANYDTALKLWIVEI

[0013] EDQQGQNQTWYANFLLGCTGYYNYDEGFMPEYPGQHQFKGTLVHPQHWPEKLDYTGKRVI

[0014] VIGSGATAITLVPSMVKGGAAHVTMLQRSPTYIASIPSIDFVYQKMRGFLSEEMAYKLTR

[0015] ARNIGMQRAVYALSQKQPKLVRKLLLKSIEMQLKGKVDMKHFTPSYNPWDQRLCVVPDGD

[0016] LFKALREGHASVETDHIEKFTETGIQLKSGKHLEADIIISATGLQIQIMGGIQGTVDGQP

[0017] IDTSEHMLYNGILISDVPNMAMIIGYINASWTLKVDVAAEYICRLLNYMDKHHYDEVIAP

[0018] TDHSEIEQDTVMGSLSAGYIRRAADVIPKQGKHAPWQVTNNYLADRKALKQAGFEDGILQFTKRDKQLERKPKLVS;

[0019] The gene sequence of the monooxygenase MOX-012 is as shown in SEQ ID NO.2:

[0020] atggataaacacattgatgttctaattgtaggagctggtatttcaggcttgggcctggct

[0021] gcgcatctttctaaaaactgcccccagcgctcatttgaaattgtagaaagacgtgaagga

[0022] atcgggggaacttgggatttattccgctatccaggtatccgctccgattcggatatgtcg ​​

[0024] atacgccagtatttacatgaggttgtggatgaatttcacctagacaggaaaattcatttt

[0025] aaacaccgagtaatttctgcgaattatgatacggcccttaaactctggatagtggaaatc

[0026] gaagaccagcaaggtcagaaccagacttggtacgctaattttctactcggctgtaccggt

[0027] tactataattatgatgaaggctttatgcctgagtaccctggacaacaccagtttaaaggt

[0028] actttggtgcatccccagcattggccagaaaagctggactatactggcaagcgagtcatt

[0029] gtgataggtagcggtgcaacagctattactctcgtaccctctatggtgaaagggggggcg

[0030] gctcatgtaaccatgctacagcgttcgcctacctatattgcttcaattccttctattgat

[0031] tttgtttatcagaaaatgcgtggttttctttcggaagagatggcttataaactgacgcgt

[0032] gcacgtaatattggtatgcagcgtgcagtctatgcactctcacagaaacaacctaaactg

[0033] gtacgtaagctccttttaaaatcgatcgaaatgcagcttaaaggtaaagtagatatgaaa

[0034] cactttactccaagctataacccatgggatcaacgcctttgtgtagtgcccgatggtgac

[0035] ctgtttaaagctttgcgtgaaggtcatgccagtgtcgagactgaccatattgaaaaattt

[0036] actgaaacaggaatccagttaaagtcaggcaagcatctggaagcagatattattatctct

[0037] gcaactggtttgcagatccagattatgggtggaatacagggaactgtagatggtcaaccg

[0038] atagatacctcagaacatatgctatacaacggtattctgatcagtgacgtacctaatatg

[0039] gcaatgattattggttatattaatgcttcttggaccttaaaagtcgatgttgcggcagag

[0040] tatatctgtcgactacttaattatatggataagcatcattacgatgaagtgatagcgcct

[0041] accgatcatagtgagattgaacaggatactgtgatgggtagcctgtccgcaggatatatt

[0042] cgtcgcgctgccgatgtaattccaaagcaggggaaacatgcaccttggcaagtgactaat

[0043] aactatctggctgatcgtaaagccctgaaacaggcgggttttgaagatggaatcttgcaatttactaaacgtgataaacagctagaacgtaaacctaaactggtttcataa;

[0044] The co-solvent is methanol, ethanol, acetonitrile, acetone, dimethyl sulfoxide, or tetrahydrofuran;

[0045] The pH value of the buffer solvent is 6 to 8.5;

[0046] The reaction temperature can be 0℃ to 30℃, more preferably 10℃ to 25℃, and more preferably room temperature.

[0047] The mass ratio of compound A to MOX-012 was 1:6; the mass ratio of compound A to GDH enzyme was 1:3; the mass ratio of compound A to NADP+ was 1:0.05; the mass ratio of compound A to buffer solution with pH=7.5 was 1:100; and the mass ratio of compound A to cosolvent was 1:5.

[0048] The reaction time can be 12-24 hours.

[0049] A preferred embodiment of the present invention is as follows: Compound A, monooxygenase, GDH enzyme, glucose, NADP+, 100 ml of buffer solution with pH=7.5 and co-solvent are stirred to obtain a solution; air is introduced to below the liquid surface, and the reaction is carried out at room temperature to prepare a sulfoxide compound.

[0050] This invention provides an enzyme-catalyzed method for synthesizing sulfoxides by sulfide oxidation. This method features mild reaction conditions, stable products, and products that are not easily over-oxidized to sulfones. It also exhibits high selectivity, with no over-oxidized impurities generated. Furthermore, it avoids the use of large amounts of oxidants and metal reagents, using air as the oxidant, thus reducing the generation of waste. The yield is over 95%, demonstrating excellent application prospects. Detailed Implementation

[0051] To further understand the present invention, the following detailed description, in conjunction with embodiments, illustrates an enzyme-catalyzed method for synthesizing sulfoxides via thioether oxidation. It should be understood that these embodiments are merely for further detailing the features of the invention and are not intended to limit the scope of the invention or the scope of the claims.

[0052] Example 1:

[0053] Weigh 1g of compound A, 6g of monooxygenase, 3g of GDH enzyme, 3g of glucose, 0.05g of NADP+, 100ml of pH 7.5 buffer solution, and 5g of dimethyl sulfoxide into a reaction flask, and then stir to obtain a solution. Purge air to below the liquid surface, and stir at room temperature for 12-24 hours until the reaction is complete. Adjust the pH to approximately 10 and then filter. Adjust the pH of the filtrate to approximately 3, and filter again after the product precipitates. Dry to obtain the product with a purity of 98% and a yield of 95%.

[0054] Example 2:

[0055] Weigh 1g of compound A, 6g of monooxygenase, 3g of GDH enzyme, 3g of glucose, 0.05g of NADP+, 100ml of pH 6.0 buffer solution, and 5g of dimethyl sulfoxide into a reaction flask, and then stir to obtain a solution. Purge air to below the liquid surface, and stir at room temperature for 12-24 hours until the reaction is complete. Adjust the pH to approximately 10 and then filter. Adjust the pH of the filtrate to approximately 3, and filter the precipitated product. Dry to obtain the product with a purity of 35% and a yield of 30%.

[0056] Example 3:

[0057] Weigh 1g of compound A, 6g of monooxygenase, 3g of GDH enzyme, 3g of glucose, 0.05g of NADP+, 100ml of pH 6.5 buffer solution, and 5g of dimethyl sulfoxide into a reaction flask, and then stir to obtain a solution. Purge air to below the liquid surface, and stir at room temperature for 12-24 hours until the reaction is complete. Adjust the pH to approximately 10 and then filter. Adjust the pH of the filtrate to approximately 3, and filter again after the product precipitates. Dry to obtain the product with a purity of 49% and a yield of 43%.

[0058] Example 4:

[0059] Weigh 1g of compound A, 6g of monooxygenase, 3g of GDH enzyme, 3g of glucose, 0.05g of NADP+, 100ml of pH 7.0 buffer solution, and 5g of dimethyl sulfoxide into a reaction flask, and then stir to obtain a solution. Purge air to below the liquid surface, and stir at room temperature for 12-24 hours until the reaction is complete. Adjust the pH to approximately 10 and then filter. Adjust the pH of the filtrate to approximately 3, and filter the precipitated product. Dry to obtain the product with a purity of 70% and a yield of 65%.

[0060] Example 5:

[0061] Weigh 1g of compound A, 6g of monooxygenase, 3g of GDH enzyme, 3g of glucose, 0.05g of NADP+, 100ml of pH 8.0 buffer solution, and 5g of dimethyl sulfoxide into a reaction flask, and then stir to obtain a solution. Purge air to below the liquid surface, and stir at room temperature for 12-24 hours until the reaction is complete. Adjust the pH to approximately 10 and then filter. Adjust the pH of the filtrate to approximately 3, and filter again after the product precipitates. Dry to obtain the product with a purity of 90% and a yield of 85%.

[0062] Example 6:

[0063] Weigh 1g of compound A, 6g of monooxygenase, 3g of GDH enzyme, 3g of glucose, 0.05g of NADP+, 100ml of pH 8.5 buffer solution, and 5g of dimethyl sulfoxide into a reaction flask, and then stir to obtain a solution. Purge air to below the liquid surface, and stir at room temperature for 12-24 hours until the reaction is complete. Adjust the pH to approximately 10 and then filter. Adjust the pH of the filtrate to approximately 3, and filter again after the product precipitates. Dry to obtain the product with a purity of 83% and a yield of 79%.

[0064] Example 7:

[0065] Weigh 1g of compound A, 6g of monooxygenase, 3g of GDH enzyme, 3g of glucose, 0.05g of NADP+, 100ml of pH 7.5 buffer solution, and 5g of methanol into a reaction flask, and then stir to obtain a solution. Purge air to below the liquid surface, and stir at room temperature for 12-24 hours until the reaction is complete. Adjust the pH to approximately 10 and then filter. Adjust the pH of the filtrate to approximately 3, and filter again after the product precipitates. Dry to obtain the product with a purity of 95% and a yield of 91%.

[0066] Example 8:

[0067] Weigh 1g of compound A, 6g of monooxygenase, 3g of GDH enzyme, 3g of glucose, 0.05g of NADP+, 100ml of pH 7.5 buffer solution, and 5g of ethanol into a reaction flask, and then stir to obtain a solution. Purge air to below the liquid surface, and stir at room temperature for 12-24 hours until the reaction is complete. Adjust the pH to approximately 10 and then filter. Adjust the pH of the filtrate to approximately 3, and filter again after the product precipitates. Dry to obtain the product; purity 90%, yield 86%.

[0068] Example 9:

[0069] Weigh 1g of compound A, 6g of monooxygenase, 3g of GDH enzyme, 3g of glucose, 0.05g of NADP+, 100ml of pH 7.5 buffer solution, and 5g of acetonitrile into a reaction flask, and then stir to obtain a solution. Purge air to below the liquid surface, and stir at room temperature for 12-24 hours until the reaction is complete. Adjust the pH to approximately 10 and then filter. Adjust the pH of the filtrate to approximately 3, and filter again after the product precipitates. Dry to obtain the product with a purity of 57% and a yield of 50%.

[0070] Example 10:

[0071] Weigh 1g of compound A, 6g of monooxygenase, 3g of GDH enzyme, 3g of glucose, 0.05g of NADP+, 100ml of pH 7.5 buffer solution, and 5g of acetone into a reaction flask, and then stir to obtain a solution. Purge air to below the liquid surface, and stir at room temperature for 12-24 hours until the reaction is complete. Adjust the pH to approximately 10 and then filter. Adjust the pH of the filtrate to approximately 3, and filter again after the product precipitates. Dry to obtain the product with a purity of 62% and a yield of 55%.

[0072] Example 11:

[0073] Weigh 1g of compound A, 6g of monooxygenase, 3g of GDH enzyme, 3g of glucose, 0.05g of NADP+, 100ml of pH 7.5 buffer solution, and 5g of tetrahydrofuran into a reaction flask, and then stir to obtain a solution. Purge air to below the liquid surface, and stir at room temperature for 12-24 hours until the reaction is complete. Adjust the pH to approximately 10 and filter. Adjust the pH of the filtrate to approximately 3, and filter again after the product precipitates. Dry to obtain the product; purity 10%, yield 8%.

[0074] This invention has illustrated its principles and implementation methods through specific embodiments. The descriptions of these embodiments are merely illustrative of the method and core ideas of this invention. Furthermore, those skilled in the art will recognize that modifications may be made to the specific implementation methods and application scope based on the principles of this invention. Therefore, the content of this specification should not be construed as limiting the invention.

[0075]

[0076]

[0077]

Claims

1. A method for preparing sulfoxide by enzyme-catalyzed oxidation of thioethers, characterized in that, Compound A, enzyme, proton donor, coenzyme, solubilizer, and buffer solution are added to a reaction flask to prepare compound B: The enzymes are monooxygenase MOX-012 and GDH enzyme; the proton donor is glucose; and the coenzyme is NADP+.

2. The preparation method according to claim 1, characterized in that, The amino acid sequence of the monooxygenase MOX-012 is shown in SEQ ID NO.1, and the gene sequence of the monooxygenase MOX-012 is shown in SEQ ID NO.

2.

3. The preparation method according to claim 1, characterized in that, The co-solvent is methanol, ethanol, acetonitrile, acetone, dimethyl sulfoxide, or tetrahydrofuran.

4. The preparation method according to claim 1, characterized in that, The pH value of the buffer solvent is 6 to 8.

5.

5. The preparation method according to claim 1, characterized in that, The mass ratio of compound A to MOX-012 is 1:

6.

6. The preparation method according to claim 1, characterized in that, The mass ratio of compound A to GDH enzyme is 1:

3.

7. The preparation method according to claim 1, characterized in that, The mass ratio of compound A to NADP+ is 1:0.

05.

8. The preparation method according to claim 1, characterized in that, The buffer solution is a buffer solution with pH = 7.5, and the mass ratio of compound A to the buffer solution with pH = 7.5 is 1:

100.

9. The preparation method according to claim 1, characterized in that, The reaction temperature is 0℃~30℃.

10. The preparation method according to claim 1, characterized in that, The mass ratio of compound A to cosolvent is 1:5.

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