METHOD FOR PHOTOCLEAVAGE OF AMIDE BONDS

Abstract

The present disclosure provides a method for photocleavage of an amide bond, the method has mild reaction conditions and can realize the cleavage of amide bonds by using light. The method comprises the following steps: subjecting 2,4-dinitrofluorobenzene to a reaction with an amino group of a substance represented by structural formula I with an α-amino acid at the end to produce a compound 1 represented by structural formula II; then under light irradiation, subjecting the compound 1 to a cleavage reaction of amide bond;

##STR00001##

Wherein, R1 is the side chain group of α-amino acid; R2 is: aryl, aliphatic hydrocarbon, —CH(R)—COOH or polypeptide.

Claims

1. A method for photocleavage of an amide bond, wherein comprising the following steps: Subjecting 2,4-dinitrofluorobenzene to a reaction with an amino group of a substance represented by structural formula I with an α-amino acid at the end to produce a compound 1 represented by structural formula II; then under light irradiation, subjecting the compound 1 to a cleavage reaction of the amide bond; ##STR00024## Wherein, R.sup.1 is the side chain group of α-amino acid; R.sup.2 is: aryl, aliphatic hydrocarbon, —CH(R)—COOH or polypeptide.

2. The method for photocleavage of the amide bond according to claim 1, wherein comprising the following steps: dissolving 2,4-dinitrofluorobenzene in an organic solvent, then dissolving the substance represented by structural formula I with an α-amino acid at the end and NaHCO.sub.3 in water, mixing the two in a reaction vessel, and stirring and refluxing at 40-100° C. under darkness for 3-10 h.

3. The method for photocleavage of the amide bond according to claim 1, wherein the light irradiation wavelength range is 250-550 nm.

4. The method for photocleavage of the amide bond according to claim 1, wherein the light irradiation time is 0.1 second to 6 h.

5. The method for photocleavage of the amide bond according to claim 2, wherein the organic solvent is an organic solvent capable of partially or completely dissolving the target lysate, i.e. compound 1.

6. The method for photocleavage of the amide bond according to claim 5, wherein the organic solvent is one or a combination of dimethyl sulfoxide, alcohol, ketone, nitrile, ether, and amide.

7. The method for photocleavage of the amide bond according to claim 6, wherein the alcohol is methanol, ethanol, butanol, ethylene glycol, n-octanol or isopropanol; the ketone is acetone, methyl ethyl ketone, methyl isobutyl ketone or cyclohexanone; the nitrile is acetonitrile, propionitrile, isobutyronitrile, butyronitrile, malononitrile, benzonitrile, benzyl cyanide, succinonitrile or glutaronitrile; the ether is diethyl ether, tetrahydrofuran, dioxane, glycol dimethyl ether, 2-methyltetrahydrofuran or diphenyl ether; and the amide is N,N-dimethylformamide or N,N-dimethylacetamide.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0018] FIG. 1 and FIG. 2 are the .sup.1H-NMR spectroscopy and high-resolution mass spectrum of compound 2;

[0019] FIG. 3 is the high-resolution mass spectrum of compound 2 after photoreaction;

[0020] FIG. 4 and FIG. 5 are high-resolution mass spectra of compound 3 and after photoreaction;

[0021] FIG. 6 and FIG. 7 are high-resolution mass spectra of compound 4 and after photoreaction;

[0022] FIG. 8 and FIG. 9 are high-resolution mass spectra of compound 5 and after photoreaction;

[0023] FIG. 10 and FIG. 11 are high-resolution mass spectra of compound 6 and after photoreaction;

[0024] FIG. 12 and FIG. 13 are high-resolution mass spectra of compound 7 and after photoreaction;

[0025] FIG. 14 and FIG. 15 are high-resolution mass spectra of compound 8 and after photoreaction;

[0026] FIG. 16 and FIG. 17 are high-resolution mass spectra of compound 9 and after photoreaction;

[0027] FIG. 18 and FIG. 19 are high-resolution mass spectra of compound 10 and after photoreaction;

[0028] FIG. 20 and FIG. 21 are high-resolution mass spectra of compound 11 and after photoreaction;

[0029] FIG. 22 and FIG. 23 are high-resolution mass spectra of compound 12 and after photoreaction;

[0030] FIG. 24 and FIG. 25 are high-resolution mass spectra of compound 13 and after photoreaction;

[0031] FIG. 26 and FIG. 27 are high-resolution mass spectra of compound 14 and after photoreaction;

[0032] FIG. 28 and FIG. 29 are high-resolution mass spectra of compound 15 and after photoreaction;

[0033] FIG. 30 and FIG. 31 are high-resolution mass spectra of compound 16 and after photoreaction;

[0034] FIG. 32 and FIG. 33 are high-resolution mass spectra of compound 17 and after photoreaction;

[0035] FIG. 34 and FIG. 35 are high-resolution mass spectra of compound 18 and after photoreaction;

[0036] FIG. 36 and FIG. 37 are high-resolution mass spectra of compound 19 and after photoreaction;

[0037] FIG. 38 and FIG. 39 are high-resolution mass spectra of compound 20 and after photoreaction;

[0038] FIG. 40 and FIG. 41 are high-resolution mass spectra of compound 21 and after photoreaction;

[0039] FIG. 42 and FIG. 43 are high-resolution mass spectra of compound 22 and after photoreaction;

DETAILED DESCRIPTION OF THE EMBODIMENTS

[0040] The present disclosure will be further described in further detail below with reference to the accompanying drawings.

Example 1

[N-(2,4-dinitrophenyl)-L-glycyl]-L-phenylalanine, GLY-PHE

[0041] ##STR00004##

[0042] L-glyto-L-phenylalanine (0.48 g, 2.16 mm) and NaHCO.sub.3 (0.3629 g, 4.32 mm) were dissolved in 25 mL deionized water, and (0.45 g, 2.41 mm) 2,4-dinitropylfluorice was dissolved in 10 mL of ethanol solution, and the two were mixed in a 100 mL round bottom flask. The mixture was stirred and refluxed at 80° C. under darkness for 5 h. After spin-drying, 0.75 g of the yellow solid product was separated by a chromatographic column using chloroform and methanol as the eluent, i.e., [N-(2,4-dinitrophenyl)-L-glycyl]-L-phenylalanine (.sup.1H-NMR spectroscopy and high-resolution mass spectrum are shown in FIG. 1 and FIG. 2). 0.01 mmol of [N-(2,4-dinitrophenyl)-L-glycinoyl]-L-phenylalanine was dissolved in 10 mL of HPLC methanol solution. The 1 mmol/L standard solution was prepared, 10 μL of the standard solution was added to 290 μL of deionized aqueous solution (pH=7), and mass spectrum data was recorded after 365 nm ultraviolet light. From the mass spectrum data, the characteristic mass spectrum peak of 5-nitro-1H-benzo[d]imidazole-3-oxide (2-a) can be clearly observed (see FIG. 3).

Example 2

[N-(2,4-dinitrophenyl)-L-alanyl]-L-alanine, ala-ala

[0043] ##STR00005##

[0044] (0.15 mM) L-alanyl-L-alanine and (0.3 mm) NaHCO.sub.3 were dissolved in 3.5 mL of deionized water, (0.18 mM) 2,4-dinitrofluorobenzene was dissolved in 2.5 mL of ethanol solution, and the two were mixed in a 25 mL flask, and the mixture was stirred at 50° C. under darkness for 4 h to obtain a yellow mixture. 5 μL of the mixture was added to 10 mL of the deionized aqueous solution (pH=7) to give the solution to be photolyzed. The characteristic peak of [N-(2,4-dinitrophenyl)-L-alanyl]-L-alanine (compound 3) can be clearly observed from the mass spectrum of the solution to be photolyzed (see FIG. 4)). 200 μL of the solution to be photolyzed was placed in a glass 96-well plate, irradiated with 365 nm ultraviolet light for 10 min, the mass spectrum data was recorded, and the characteristic mass spectrum peak of 2-methyl-5-nitro-1H-benzo[d]imidazole-3-oxide (3-a) can be clearly observed (see FIG. 5).

Example 3

[N-(2,4-dinitrophenyl)-L-alanyl]-L-glycine, ala-gly

[0045] ##STR00006##

[0046] In the same conditions as in Example 2, L-alanyl-L-glycine was reacted with 2,4-dinitrofluorobenzene, and the characteristic peak of [N-(2,4-dinitrophenyl)-L-alanyl]-L-glycine (compound 3) can be clearly observed on the mass spectrum data after the reaction (see FIG. 6). Under the same conditions as in Example 2, the compound 3 was subjected to a photocleavage reaction, the characteristic peak of 2-methyl-5-nitro-1H-benzo[d]imidazole-3-oxide (3-a) can be clearly observed on the mass spectrum after the photoreaction (see FIG. 7).

Example 4

[N-(2,4-dinitrophenyl)-L-histidyl]-L-leucine His-leu

[0047] ##STR00007##

[0048] Under the same conditions as in Example 2, L-histidyl-L-leucine was reacted with 2,4-dinitrofluorobenzene, and the characteristic peaks of [N-(2, 4-dinitrophenyl)-L-histaminoyl]-L-leucine (compound 5) and [N,N′-bis(2, 4-dinitrophenyl) L-histaminoyl]-L-leucine (compound 5′) could be clearly observed on the mass spectrum data after the reaction (See FIG. 8). Under the same conditions as in Example 2, compound 5 and compound 5′ were subjected to photocleavage reaction, and the characteristic peaks of 2-[1H-imidazol-4-yl)methyl]-5-nitro-1H-benzo[d]imidazole-3-oxide (5-a) and 2-[1-(2,4-dinitrophenyl)-1H-imidazol-4-yl)methyl)-5-nitro-1H-benzo[d]imidazole-3-oxide (5-b) could be clearly observed on the mass spectrum data after the reaction (see FIG. 9).

Example 5

[N-(2,4-dinitrophenyl)-L-alanyl]-L-phenylalanine Ala-phe

[0049] ##STR00008##

[0050] Under the same conditions as in Example 2, L-alanyl-L-phenylalanine was reacted with 2,4-dinitrofluorobenzene, the characteristic peak of [N-(2,4-dinitrophenyl)-L-alanyl]-L-phenylalanine (compound 6) can be clearly observed on the mass spectrum data after the reaction (see FIG. 10). Under the same conditions as in Example 2, compound 6 was subjected to a photocleavage reaction, the characteristic peak of 2-methyl-5-nitro-1H-benzo[d]imidazole-3-oxide (6-a) can be clearly observed on the mass spectrum after the photoreaction (see FIG. 11).

Example 6

[N-(2,4-dinitrophenyl)-L-alanyl]-L-glutamine Ala-gln

[0051] ##STR00009##

[0052] Under the same conditions as in Example 2, L-alanyl-L-glutamine was reacted with 2,4-dinitrofluorobenzene, the characteristic peak of [N-(2,4-dinitrophenyl)-L-alanyl]-L-glutamine (compound 7) can be clearly observed on the mass spectrum data after the reaction (see FIG. 12). Under the same conditions as in Example 2, [N-(2,4-dinitrophenyl)-L-alanyl]-L-glutamine was subjected to a photocleavage reaction, the characteristic peak of 2-methyl-5-nitro-1H-benzo[d]imidazole-3-oxide (7-a) can be clearly observed on the mass spectrum after the photoreaction (see FIG. 13).

Example 7

[N-(2,4-dinitrophenyl)-L-alanyl]-L-leucine Ala-leu

[0053] ##STR00010##

[0054] Under the same conditions as in Example 2, L-alanyl-L-leucine was reacted with 2,4-dinitrofluorobenzene, the characteristic peak of [N-(2,4-dinitrophenyl)-L-alanyl]-L-leucine (compound 8) can be clearly observed on the mass spectrum data after the reaction (see FIG. 14). Under the same conditions as in Example 2, [N-(2,4-dinitrophenyl)-L-alanyl]-L-leucine was subjected to a photocleavage reaction, the characteristic peak of 2-methyl-5-nitro-1H-benzo[d]imidazole-3-oxide (8-a) can be clearly observed on the mass spectrum after the photoreaction (see FIG. 15).

Example 8

[N-(2,4-dinitrophenyl)-L-leucyl]-L-isoleucine leu-ile

[0055] ##STR00011##

[0056] Under the same conditions as in Example 2, L-leucyl-L-isoleucine was reacted with 2,4-dinitrofluorobenzene, the characteristic peak of [N-(2,4-dinitrophenyl)-L-leucyl]-L-isoleucine (compound 9) can be clearly observed on the mass spectrum data after the reaction (see FIG. 16). Under the same conditions as in Example 2, [N-(2,4-dinitrophenyl)-L-leucyl]-L-isoleucine was subjected to a photocleavage reaction, the characteristic peak of 2-isobutyl-5-nitro-1H-benzo[d]imidazole-3-oxide (9-a) can be clearly observed on the mass spectrum after the photoreaction (see FIG. 17).

Example 9

[N-(2,4-dinitrophenyl)-L-leucyl]-L-phenylalanine leu-phe

[0057] ##STR00012##

[0058] Under the same conditions as in Example 2, L-leucyl-L-phenylalanine was reacted with 2,4-dinitrofluorobenzene, the characteristic peak of [N-(2,4-dinitrophenyl)-L-leucyl]-L-phenylalanine (compound 10) can be clearly observed on the mass spectrum data after the reaction (see FIG. 18). Under the same conditions as in Example 2, [N-(2,4-dinitrophenyl)-L-leucyl]-L-phenylalanine was subjected to a photocleavage reaction, the characteristic peak of 2-isobutyl-5-nitro-1H-benzo[d]imidazole-3-oxide (10-a) can be clearly observed on the mass spectrum after the photoreaction (see FIG. 19).

Example 10

[N-(2,4-dinitrophenyl)-L-leucyl]-L-leucine leu-leu

[0059] ##STR00013##

[0060] Under the same conditions as in Example 2, L-leucyl-L-leucine was reacted with 2,4-dinitrofluorobenzene, the characteristic peak of [N-(2,4-dinitrophenyl)-L-leucyl]-L-leucine (compound 11) can be clearly observed on the mass spectrum data after the reaction (see FIG. 20). Under the same conditions as in Example 2, [N-(2,4-dinitrophenyl)-L-leucyl]-L-leucine was subjected to a photocleavage reaction, the characteristic peak of 2-isobutyl-5-nitro-1H-benzo[d]imidazole-3-oxide (11-a) can be clearly observed on the mass spectrum after the photoreaction (see FIG. 21).

Example 11

[N-(2,4-dinitrophenyl)-L-phenylalanyl]-L-leucine phe-leu

[0061] ##STR00014##

[0062] Under the same conditions as in Example 2, L-phenylalanyl-L-leucine was reacted with 2,4-dinitrofluorobenzene, the characteristic peak of [N-(2,4-dinitrophenyl)-L-phenylalanyl]-L-leucine (compound 12) can be clearly observed on the mass spectrum data after the reaction (see FIG. 22). Under the same conditions as in Example 2, [N-(2,4-dinitrophenyl)-L-phenylalanyl]-L-leucine was subjected to a photocleavage reaction, the characteristic peak of 2-benzyl-5-nitro-1H-benzo[d]imidazole-3-oxide (12-a) can be clearly observed on the mass spectrum after the photoreaction (see FIG. 23).

Example 12

[N-(2,4-dinitrophenyl)-L-phenylalanyl]-L-alanine phe-ala

[0063] ##STR00015##

[0064] Under the same conditions as in Example 2, L-phenylalanyl-L-alanine was reacted with 2,4-dinitrofluorobenzene, the characteristic peak of [N-(2,4-dinitrophenyl)-L-phenylalanyl]-L-alanine (compound 13) can be clearly observed on the mass spectrum data after the reaction (see FIG. 24). Under the same conditions as in Example 2, [N-(2,4-dinitrophenyl)-L-phenylalanyl]-L-alanine was subjected to a photocleavage reaction, the characteristic peak of 2-benzyl-5-nitro-1H-benzo[d]imidazole-3-oxide (13-a) can be clearly observed on the mass spectrum after the photoreaction (see FIG. 25).

Example 13

[N-(2,4-dinitrophenyl)-L-phenylalanyl]-L-phenylalanine phe-phe

[0065] ##STR00016##

[0066] Under the same conditions as in Example 2, L-phenylalanyl-L-phenylalanine was reacted with 2,4-dinitrofluorobenzene, the characteristic peak of [N-(2,4-dinitrophenyl)-L-phenylalanyl]-L-phenylalanine (compound 13) can be clearly observed on the mass spectrum data after the reaction (see FIG. 26). Under the same conditions as in Example 2, [N-(2,4-dinitrophenyl)-L-phenylalanyl]-L-phenylalanine was subjected to a photocleavage reaction, the characteristic peak of 2-benzyl-5-nitro-1H-benzo[d]imidazole-3-oxide (13-a) can be clearly observed on the mass spectrum after the photoreaction (see FIG. 27).

Example 14

[N-(2,4-dinitrophenyl)-L-leucyl]-L-glycine LEU-GLY

[0067] ##STR00017##

[0068] Under the same conditions as in Example 2, L-leucyl-L-glycine was reacted with 2,4-dinitrofluorobenzene, the characteristic peak of [N-(2,4-dinitrophenyl)-L-leucyl]-L-glycine (compound 15) can be clearly observed on the mass spectrum data after the reaction (see FIG. 28). Under the same conditions as in Example 2, [N-(2,4-dinitrophenyl)-L-leucyl]-L-glycine was subjected to a photocleavage reaction, the characteristic peak of 2-isobutyl-5-nitro-1H-benzo[d]imidazole-3-oxide (15-a) can be clearly observed on the mass spectrum data after the reaction (see FIG. 29).

Example 15

[N-(2,4-dinitrophenyl)-L-leucyl]-L-tyrosine LEU-tyr

[0069] ##STR00018##

[0070] Under the same conditions as in Example 2, L-leucyl-L-tyrosine was reacted with 2,4-dinitrofluorobenzene, under the same conditions as in Example 2, L-leucyl-L-tyrosine was reacted with 2,4-dinitrofluorobenzene, the characteristic peak of [N-(2,4-dinitrophenyl)-L-leucyl]-L-tyrosine (compound 16) can be clearly observed on the mass spectrum data after the reaction (see FIG. 30). Under the same conditions as in Example 2, [N-(2,4-dinitrophenyl)-L-leucyl]-L-tyrosine was subjected to a photocleavage reaction, the characteristic peak of 2-isobutyl-5-nitro-1H-benzo[d]imidazole-3-oxide (16-a) can be clearly observed on the mass spectrum after the photoreaction (see FIG. 31).

Example 16

[N-(2,4-dinitrophenyl)-L-leucyl]-L-valine leu-val

[0071] ##STR00019##

[0072] Under the same conditions as in Example 2, L-leucyl-L-valine was reacted with 2,4-dinitrofluorobenzene, the characteristic peak of [N-(2,4-dinitrophenyl)-L-leucyl]-L-valine (compound 17) can be clearly observed on the mass spectrum data after the reaction (see FIG. 32). Under the same conditions as in Example 2, [N-(2,4-dinitrophenyl)-L-leucyl]-L-valine was subjected to a photocleavage reaction, the characteristic peak of 2-isobutyl-5-nitro-1H-benzo[d]imidazole-3-oxide (17-a) can be clearly observed on the mass spectrum after the photoreaction (see FIG. 33).

Example 17

[N-(2,4-dinitrophenyl)-L-methionyl]-L-glycine MET-GLY

[0073] ##STR00020##

[0074] Under the same conditions as in Example 2, L-methionyl-L-glycine was reacted with 2,4-dinitrofluorobenzene, the characteristic peak of [N-(2,4-dinitrophenyl)-L-methionyl]-L-glycine (compound 18) can be clearly observed on the mass spectrum data after the reaction (see FIG. 34). Using deionized water with pH=6 instead of deionized water with pH=7, under the same conditions as in Example 2, [N-(2,4-dinitrophenyl)-L-methionyl]-L-glycine was subjected to a photocleavage reaction, the characteristic peak of 2-[2-(methylthio)ethyl]-5-nitro-1H-benzo[d]imidazole-3-oxide (18-a) can be clearly observed on the mass spectrum data after the reaction (see FIG. 35).

Example 18

[N-(2,4-dinitrophenyl)-L-phenylalanyl]-L-glycine phe-GLY

[0075] ##STR00021##

[0076] Under the same conditions as in Example 2, L-phenylalanyl-L-glycine was reacted with 2,4-dinitrofluorobenzene, the characteristic peak of [N-(2,4-dinitrophenyl)-L-phenylalanyl]-L-glycine (compound 19) can be clearly observed on the mass spectrum data after the reaction (see FIG. 36). Under the same conditions as in Example 2, [N-(2,4-dinitrophenyl)-L-phenylalanyl]-L-glycine was subjected to a photocleavage reaction, the characteristic peak of 2-benzyl-5-nitro-1H-benzo[d]imidazole-3-oxide (19-a) can be clearly observed on the mass spectrum after the photoreaction (see FIG. 37).

Example 19

[N-(2,4-dinitrophenyl)-L-glycyl]-L-glutamic acid GLY-glu

[0077] ##STR00022##

[0078] Under the same conditions as in Example 2, L-glycyl-L-glutamic acid was reacted with 2,4-dinitrofluorobenzene, the characteristic peak of [N-(2,4-dinitrophenyl)-L-glycyl]-L-glutamic acid (compound 20) can be clearly observed on the mass spectrum data after the reaction (see FIG. 38). Using deionized water of pH=6 instead of deionized water of pH=7, under the same conditions as in Example 2, [N-(2,4-dinitrophenyl)-L-glycyl]-L-glutamic acid was subjected to a photocleavage reaction, and the characteristic peak of 5-nitro-1H-benzo[d]imidazole-3-oxide (20-a) can be clearly observed in the mass spectrum after the photoreaction (see FIG. 39).

Example 20

[N-(2,4-dinitrophenyl)-L-valinyl]-L-alanine val-ala

[0079] ##STR00023##

[0080] Under the same conditions as in Example 2, L-valinyl-L-alanine was reacted with 2,4-dinitrofluorobenzene, the characteristic peak of [N-(2,4-dinitrophenyl)-L-valinyl]-L-alanine (compound 21) can be clearly observed on the mass spectrum data after the reaction (see FIG. 40). Using deionized water with pH=6 instead of deionized water with pH=7, under the same conditions as in Example 2, [N-(2,4-dinitrophenyl)-L-valinyl]-L-alanine was subjected to a photocleavage reaction, the characteristic peak of 2-isopropyl-5-nitro-1H-benzo[d]imidazole-3-oxide (21-a) can be clearly observed on the mass spectrum after the photoreaction (see FIG. 41).

Example 21

[N-(2,4-dinitrophenyl)-L-prolyl]-glycine pro-gly

[0081] Under the same conditions as in Example 2, L-prolyl-L-glycine was reacted with 2,4-dinitrofluorobenzene, the characteristic peak of [N-(2,4-dinitrophenyl)-L-prolyl]-L-glycine (compound 22) can be clearly observed on the mass spectrum data after the reaction (see FIG. 42). Using deionized water with pH=6 instead of deionized water with pH=7, under the same conditions as in Example 2, [N-(2,4-dinitrophenyl)-L-prolyl]-L-glycine was subjected to a photocleavage reaction, the characteristic peak of 6-nitro-2,3-dihydro-1H-benzo[d]pyrrole[1,2-a]imidazole 4-oxide (22-a) can be clearly observed on the mass spectrum data after the reaction (see FIG. 43).