Preparation method of biotinylated amino acid derivative

Abstract

The invention belongs to the technical field of preparation of amino acid derivatives, and relates to a preparation method of a biotinylated amino acid derivative, which is characterized by comprising the following steps: adding amino acid and alkali into biotin D-biotin, stirring, adding a catalyst and a solvent, stirring to react, pumping the solvent, extracting, and purifying to obtain the biotinylated amino acid derivative. The catalyst is 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide and 1-hydroxybenzotriazole, and the catalyst is a mixture of a catalyst and a catalyst. According to the method, metal catalysis or reagents such as high-activity acyl chloride are not needed, amino acidification of biotin D-biotin is achieved under the action of alkali under mild conditions, and the method is easy to operate, mild in reaction condition, wide in substrate application range, low in raw material cost and easy to obtain, high in product selectivity and wide in market prospect.

Description

Technical field:

[0001] The invention belongs to the technical field of preparation of amino acid derivatives, and relates to a method for preparing biotinylated amino acid derivatives. The method uses biotin D-biotin as a raw material and reacts with various amino acids under alkaline conditions to obtain corresponding biotinylated amino acid derivatives. Background technology:

[0002] Biotin derivatives are an important class of organic functional molecules, which are widely used in the fields of health medicine and biomaterials. For example, biotin derivatives NHS-biotin, Sulfo-NHS-biotin, NHS-LC-biotin haid Sulfo-NHS-LC-Biotin for labeling protein amino groups; activated biotin for labeling protein sulfhydryl groups: Biotin-BMCC, Biotin-HPDP; Biotin-Hydrazide haid Biotin-LC-Hydrazide, Biocytin Hydrazide for labeling protein carbonyl groups, etc. Therefore, the development of methods for preparing biotin derivatives is of great significance and research value for the development of new drugs and biological functional molecules. At present, many best-selling marker molecules on the market contain biotin structural units. Combining biotin with amino acids is an effective means of functionalizing amino acids for labeling amino acids. Functionalized amino acids retain the functions of amino acids and have the fluorescence tracking function of the new fragment biotin D-biotin. The combination of the two is expected to have important significance and practical value in the fields of health care and new drug development such as proteins.

[0003]

[0004] At present, the most commonly used method for synthesizing amides is to react acyl chlorides with amines to obtain amides. This method first requires the reaction of carboxylic acid and dithionyl chloride to obtain acyl chlorides, such as [Chem. Eur. J. 2008, 14, 9864–9867, [Org. Biomol. Chem., 2015, 13, 7803–7812], etc. The reaction rate of acyl chlorides and amines is fast, but because the acyl chlorides are too active, the functional group compatibility is poor; the secondary amides obtained can continue to react with acyl chlorides, with many by-products, low yields, and poor functional group selectivity. Amides can also be obtained by using transition metals to catalyze the imine reaction. This method requires the use of expensive metal catalysts, such as dimerized rhodium acetate as a catalyst [Angew. Chem. Int. Ed. 2005, 44, 3096–3099]. The participation of metals increases the difficulty of subsequent separation and purification, especially in pharmaceutical production, where the requirement for low metal content greatly reduces the practical value of the synthesis method.

[0005] Therefore, it is extremely urgent to provide a simple, efficient and metal-free method for amino acidification of biotin D-biotin, which will have important research significance and practical value in the fields of health care and new drug development. Summary of the invention:

[0006] The purpose of the present invention is to overcome the problems existing in the prior art and provide a method for preparing functional amino acid derivatives by using biotin D-biotin and amino acids as raw materials and organic bases or inorganic bases as alkaline reagents to realize the amidation reaction of D-biotin, that is, using biotin D-biotin and amino acids as raw materials to obtain corresponding biotin derivatives. The method is simple to operate, has a high yield, a wide range of substrate applications, mild reaction conditions, and high selectivity.

[0007] In order to achieve the above-mentioned purpose, the present invention provides a method for preparing biotinylated amino acid derivatives. With biotin D-biotin and various amino acids as substrates, in the presence of an organic base or an apolar base, using EDC and HOBt as co-catalysts, biotin D-biotin is subjected to a directional amino acidization reaction in an organic solvent to obtain amino acidized biotin derivatives. The specific method is as follows: firstly, biotin D-biotin is added to a reaction tube, and then, under the protection of an inert gas at -78°C, amino acids and bases are first added, and stirred at room temperature for 0.5h; then, under the protection of an inert gas at -78°C, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC), 1-hydroxybenzotriazole (HOBt) and a solvent are added, and the reaction is stirred for 1-48h, the solvent is drained, and the reaction is washed with n-hexane, extracted with dichloromethane, and purified by a chemical method to obtain the corresponding biotin derivatives.

[0008] The method for preparing biotin derivatives of the present invention has the following general reaction formula:

[0009]

[0010] Where R 1 is hydrogen, methyl, ethyl, tert-butyl, methoxy, methylthio, trifluoromethyl, trifluoromethoxy, amino, substituted amino, vinyl, styryl, phenylethynyl, phenyl, acylamino, benzamido, hydroxy, substituted hydroxy, thiol, substituted thiol, pinacol boronyl, oxazolyl, pyrazolyl, pyridyl, thienyl, thiazolyl, furanyl, pyrrolyl, morpholinyl, piperidinyl, fluorine, chlorine, bromine, iodine functional group or cyclic group; R 2It is hydrogen, (substituted) methyl, ethyl, tert-butyl, methoxy, methylthio, trifluoromethyl, trifluoromethoxy, amino, substituted amino, vinyl, styryl, phenylethynyl, phenyl, acylamino, benzamido, hydroxy, hydroxymethyl, thiol, thiolmethyl, pinacol boronyl, oxazolyl, pyrazolyl, pyridyl, thienyl, thiazolyl, furanyl, pyrrolyl, morpholinyl, piperidinyl, fluorine, chlorine, bromine, iodine functional group or cyclic group.

[0011] The chemical structural formula of the EDC compound of the present invention is as follows:

[0012] The chemical structural formula of the HOBt compound of the present invention is as follows:

[0013] The chemical structural formula of biotin D-biotin of the present invention is as follows:

[0014]

[0015] The amino acid of the present invention is at least one of glycine, alanine, valine, leucine, isoleucine, methionine (methionine), proline, tryptophan, serine, tyrosine, cysteine, phenylalanine, asparagine, glutamine, threonine, aspartic acid, glutamic acid, lysine, arginine, histidine, selenocysteine ​​or pyrrolysine.

[0016] The solvent described in the present invention is a polar solvent or a non-polar solvent, and is selected from at least one of ethanol, methanol, ethyl acetate, ether, dioxane, tetrahydrofuran, ethylene glycol dimethyl ether, 1,2-dichloroethane, dichloromethane, acetonitrile, 1,4-dioxane, benzene, toluene, xylene, trifluorotoluene, N,N-dimethylformamide, N,N-dimethylacetamide, and dimethyl sulfoxide.

[0017] The inert gas described in the present invention is nitrogen or argon.

[0018] The reaction temperature of the present invention is between 0°C and 100°C.

[0019] The reaction time of the present invention is 1-48 hours.

[0020] The molar ratio of biotin, alkali, EDC, HOBt and amino acid is 1:(1-10):(1-10):(1-10):(1-10).

[0021] The base of the present invention is an inorganic base or an organic base. The inorganic base is lithium hydroxide, sodium hydroxide, potassium hydroxide, sodium carbonate, sodium bicarbonate, potassium carbonate, potassium bicarbonate, cesium carbonate, calcium carbonate, sodium phosphate, disodium hydrogen phosphate or sodium dihydrogen phosphate. The organic base is sodium methoxide, potassium ethoxide, potassium tert-butoxide, butyl lithium, phenyl lithium, lithium diisopropylamide (LDA), lithium hexamethyldisilazide (LiHMDS), triethylamine or trimethylamine.

[0022] Compared with the existing methods, the present invention has the following advantages: no metal catalysis or highly active reagents such as acyl chloride are required, and the amino acidization of biotin D-biotin is achieved under mild conditions under the action of alkali. The method is simple to operate, has mild reaction conditions, a wide range of substrate applications, cheap and readily available raw materials, high product selectivity, and broad market prospects. Description of the drawings:

[0023] Figure 1 This is a product status diagram of Example 1 involved in the present invention.

[0024] Figure 2 This is the nuclear magnetic resonance spectrum of the product of Example 1 of the present invention.

[0025] Figure 3 This is a high-resolution mass spectrum of the product of Example 1 of the present invention. Specific implementation method:

[0026] The present invention is further described in detail below by way of examples.

[0027] Embodiment 1:

[0028] The present invention relates to a method for synthesizing and preparing a biotinylated amino acid derivative, wherein the amino acid selected is β-alanine, and the specific steps are as follows:

[0029]

[0030] In the reaction tube, add 1mmol biotin D-biotin, under nitrogen protection, add 89mg β-alanine and 101mg triethylamine at -78℃, and stir at room temperature for 0.5h; then under nitrogen protection, add 157mg EDC, 135mg HOBt and 2mL DCM (dichloromethane) at -78℃, and then stir and react at 35℃ for 24h; after the reaction is completed, drain the solvent, wash with n-hexane, extract with dichloromethane, spin dry the solvent, and chromatograph on a silica gel column (the eluent is a mixture of petroleum ether and ethyl acetate in a volume ratio of 1:1) to obtain 252mg of the product with a yield of 80%. The product state, product nuclear magnetic resonance and product high-resolution mass spectrometry characterization results are shown as follows Figure 1-3 shown.

[0031] Embodiment 2:

[0032] The present invention relates to a method for synthesizing and preparing a biotinylated amino acid derivative, wherein the amino acid substrate selected isoserine, and the specific steps are as follows:

[0033]

[0034] In a reaction tube, 1 mmol biotin was added, and under nitrogen protection, 105 mg isoserine and 101 mg triethylamine were added at -78°C, and the mixture was stirred at room temperature for 0.5 h; then, under nitrogen protection, 157 mg EDC, 135 mg HOBt and 2 mL DCM were added at -78°C, and the mixture was stirred and reacted at 35°C for 24 h. After the reaction was completed, the solvent was drained, the mixture was washed with n-hexane, extracted with dichloromethane, and the solvent was dried by spin drying. After silica gel column chromatography (the eluent was a mixture of petroleum ether and ethyl acetate in a volume ratio of 1:1), 248 mg of the product was obtained, and the yield was 75%.

[0035] Embodiment 3:

[0036] The present invention relates to a method for synthesizing and preparing a biotinylated amino acid derivative, wherein the amino acid substrate selected is glycine, and the specific steps are as follows:

[0037]

[0038] In a reaction tube, 1 mmol biotin was added, and under nitrogen protection, 75 mg glycine and 101 mg triethylamine were added at -78°C, and the mixture was stirred at room temperature for 0.5 h. Under nitrogen protection, 157 mg EDC, 135 mg HOBt and 2 mL DCM were added at -78°C, and then the mixture was stirred and reacted at 35°C for 24 h. After the reaction was completed, the solvent was drained, the mixture was washed with n-hexane, extracted with dichloromethane, the solvent was spin-dried, and the mixture was chromatographed on a silica gel column (the eluent was a mixture of petroleum ether and ethyl acetate in a volume ratio of 1:1) to obtain 210 mg of the product with a yield of 70%.

[0039] Embodiment 4:

[0040] The present invention relates to a method for synthesizing and preparing a biotinylated amino acid derivative, wherein the amino acid substrate selected is L-alanine, and the specific steps are as follows:

[0041]

[0042] In a reaction tube, add 1mmol biotin, under nitrogen protection, add 89mg L-alanine and 101mg triethylamine at -78°C, and stir at room temperature for 0.5h. Under nitrogen protection, add 157mg EDC, 135mg HOBt, and 2mL DCM at -78°C, stir at 35°C for 24h, drain the solvent, wash with n-hexane, extract with dichloromethane, spin dry the solvent, and chromatograph on a silica gel column (the eluent is a mixture of petroleum ether and ethyl acetate in a volume ratio of 1:1) to obtain 255mg of the product with a yield of 81%.

[0043] Embodiment 5:

[0044] The present invention relates to a method for synthesizing and preparing a biotinylated amino acid derivative, wherein the amino acid substrate selected is serine, and the specific steps are as follows:

[0045]

[0046] In a reaction tube, 1 mmol biotin was added, and under nitrogen protection, 105 mg serine and 101 mg triethylamine were added at -78°C, and the mixture was stirred at room temperature for 0.5 h. Under nitrogen protection, 157 mg EDC, 135 mg HOBt and 2 mL DCM were added at -78°C, and then the mixture was stirred and reacted at 35°C for 24 h. After the reaction was completed, the solvent was drained, the mixture was washed with n-hexane, extracted with dichloromethane, the solvent was spin-dried, and the product was chromatographed on a silica gel column (the eluent was a mixture of petroleum ether and ethyl acetate in a volume ratio of 1:1) to obtain 166 mg of the product with a yield of 50%.

[0047] Embodiment 6:

[0048] The present invention relates to a method for synthesizing and preparing a biotinylated amino acid derivative, wherein the amino acid substrate selected is tert-leucine, and the specific steps are as follows:

[0049]

[0050] In a reaction tube, 1 mmol biotin was added, and under nitrogen protection, 131 mg tert-leucine and 101 mg triethylamine were added at -78°C, and the mixture was stirred at room temperature for 0.5 h. Under nitrogen protection, 157 mg EDC, 135 mg HOBt and 2 mL DCM were added at -78°C, and then the mixture was stirred and reacted at 35°C for 24 h. After the reaction was completed, the solvent was drained, the mixture was washed with n-hexane, extracted with dichloromethane, the solvent was spin-dried, and the mixture was chromatographed on a silica gel column (the eluent was a mixture of petroleum ether and ethyl acetate in a volume ratio of 1:1) to obtain 242 mg of the product with a yield of 68%.

[0051] Embodiment 7:

[0052] The present invention relates to a method for synthesizing and preparing a biotinylated amino acid derivative, wherein the amino acid substrate selected is threonine, and the specific steps are as follows:

[0053]

[0054] In a reaction tube, 1 mmol biotin was added, and under nitrogen protection, 119 mg threonine and 101 mg triethylamine were added at -78°C, and the mixture was stirred at room temperature for 0.5 h. Under nitrogen protection, 157 mg EDC, 135 mg HOBt and 2 mL DCM were added at -78°C, and then the mixture was stirred and reacted at 35°C for 24 h. After the reaction was completed, the solvent was drained, the mixture was washed with n-hexane, extracted with dichloromethane, the solvent was spin-dried, and the mixture was chromatographed on a silica gel column (the eluent was a mixture of petroleum ether and ethyl acetate in a volume ratio of 1:1) to obtain 207 mg of the product with a yield of 60%.

[0055] Embodiment 8:

[0056] The present invention relates to a method for synthesizing and preparing a biotinylated amino acid derivative, wherein the amino acid substrate selected is β-hydroxynorvaline, and the specific steps are as follows:

[0057]

[0058] In a reaction tube, 1 mmol biotin was added, and under nitrogen protection, 133 mg β-hydroxynorvaline and 101 mg triethylamine were added at -78°C, and the mixture was stirred at room temperature for 0.5 h; under nitrogen protection, 157 mg EDC, 135 mg HOBt and 2 mL DCM were added at -78°C, and then the mixture was stirred and reacted at 35°C for 24 h. After the reaction was completed, the solvent was drained, the mixture was washed with n-hexane, extracted with dichloromethane, the solvent was spin-dried, and the product was chromatographed on a silica gel column (the eluent was a mixture of petroleum ether and ethyl acetate in a volume ratio of 1:1) to obtain 219 mg of the product with a yield of 61%.

[0059] Embodiment 9:

[0060] The present invention relates to a method for synthesizing and preparing a biotinylated amino acid derivative. The amino acid substrate selected is methionine. The specific reaction equation and steps are as follows:

[0061]

[0062] In a reaction tube, 1 mmol biotin was added, and under nitrogen protection, 146 mg methionine and 101 mg triethylamine were added at -78°C, and the mixture was stirred at room temperature for 0.5 h. Under nitrogen protection, 157 mg EDC, 135 mg HOBt and 2 mL DCM were added at -78°C, and then the mixture was stirred and reacted at 35°C for 24 h. After the reaction was completed, the solvent was drained, the mixture was washed with n-hexane, extracted with dichloromethane, the solvent was spin-dried, and the mixture was chromatographed on a silica gel column (the eluent was a mixture of petroleum ether and ethyl acetate in a volume ratio of 1:1) to obtain 270 mg of the product with a yield of 72%.

[0063] Embodiment 10:

[0064] The present invention relates to a method for synthesizing and preparing a biotinylated amino acid derivative. The amino acid substrate selected is cysteine. The specific reaction equation and steps are as follows:

[0065]

[0066] In a reaction tube, 1 mmol biotin was added, and under nitrogen protection, 121 mg cysteine ​​and 101 mg triethylamine were added at -78°C, and the mixture was stirred at room temperature for 0.5 h. Under nitrogen protection, 157 mg EDC, 135 mg HOBt and 2 mL DCM were added at -78°C, and then the mixture was stirred and reacted at 35°C for 24 h. After the reaction was completed, the solvent was drained, the mixture was washed with n-hexane, extracted with dichloromethane, the solvent was spin-dried, and the product was chromatographed on a silica gel column (the eluent was a mixture of petroleum ether and ethyl acetate in a volume ratio of 1:1) to obtain 191 mg of the product with a yield of 55%.

[0067] Embodiment 11:

[0068] The present invention relates to a method for synthesizing and preparing a biotinylated amino acid derivative. The amino acid substrate selected is histidine. The specific reaction equation and steps are as follows:

[0069]

[0070] In a reaction tube, 1 mmol biotin was added, and under nitrogen protection, 155 mg histidine and 101 mg triethylamine were added at -78°C, and the mixture was stirred at room temperature for 0.5 h. Under nitrogen protection, 157 mg EDC, 135 mg HOBt and 2 mL DCM were added at -78°C, and then the mixture was stirred and reacted at 35°C for 24 h. After the reaction was completed, the solvent was drained, the mixture was washed with n-hexane, extracted with dichloromethane, the solvent was spin-dried, and the product was chromatographed on a silica gel column (the eluent was a mixture of petroleum ether and ethyl acetate in a volume ratio of 1:1) to obtain 186 mg of the product with a yield of 49%.

[0071] Embodiment 12:

[0072] Compared with Example 1, in this example, triethylamine is replaced by potassium carbonate, and the specific steps are as follows:

[0073]

[0074] In the reaction tube, add 1mmol biotin, under nitrogen protection, add 146mg β-alanine and 138mg potassium carbonate at -78℃, and stir at room temperature for 0.5h. Under nitrogen protection, add 157mg EDC, 135mg HOBt and 2mL DCM at -78℃, stir at 35℃ for 24h, drain the solvent, wash with n-hexane, extract with dichloromethane, spin dry the solvent, and chromatograph on a silica gel column (the eluent is a mixture of petroleum ether and ethyl acetate in a volume ratio of 1:1), and obtain 258mg of the product with a yield of 82%. This shows that the reaction can also be achieved by replacing triethylamine with potassium carbonate.

[0075] Comparative Example 1:

[0076] Compared with Example 1, the solvent DCM was replaced with THF in this comparative example, and the specific steps were as follows:

[0077]

[0078] In a reaction tube, add 1mmol biotin, under nitrogen protection, add 146mg β-alanine and 101mg triethylamine at -78°C, and stir at room temperature for 0.5h. Under nitrogen protection, add 157mg EDC, 135mg HOBt, and 2mL THF (tetrahydrofuran) at -78°C, stir at 35°C for 24h, drain the solvent, wash with n-hexane, extract with dichloromethane, spin dry the solvent, and chromatograph on a silica gel column (the eluent is a mixture of petroleum ether and ethyl acetate in a volume ratio of 1:1) to obtain 236mg of the product with a yield of 75%.

[0079] Comparative Example 2:

[0080] Compared with Example 1, the reaction temperature in this comparative example is set to 100° C., and the specific steps are as follows:

[0081]

[0082] In a reaction tube, 1 mmol biotin was added, and under nitrogen protection, 146 mg β-alanine and 101 mg triethylamine were added at -78°C, and the mixture was stirred at room temperature for 0.5 h. Under nitrogen protection, 157 mg EDC, 135 mg HOBt and 2 mL DCM were added at -78°C, and the mixture was stirred and reacted at 100°C for 24 h. The solvent was drained, and the mixture was washed with n-hexane and extracted with dichloromethane. The solvent was spin-dried and chromatographed on a silica gel column (the eluent was a mixture of petroleum ether and ethyl acetate in a volume ratio of 1:1) to obtain 252 mg of the product with a yield of 80%.

[0083] Comparative Example 3:

[0084] Compared with Example 1, this comparative example did not add triethylamine to the reaction raw materials, and the specific steps were as follows:

[0085]

[0086] In the reaction tube, add 1mmol biotin, under nitrogen protection, add 146mg β-alanine at -78℃, and stir at room temperature for 0.5h. Under nitrogen protection, add 157mg EDC, 135mg HOBt, and 2mL DCM at -78℃, stir at 35℃ for 24h, drain the solvent, wash with n-hexane, extract with dichloromethane, spin dry the solvent, and chromatograph on a silica gel column (the eluent is a mixture of petroleum ether and ethyl acetate in a volume ratio of 1:1), and obtain 0mg of the product with a yield of 0%. This shows that without the participation of a base, biotin and amino acids cannot react, and no amino acid derivatives can be obtained.

[0087] Comparative Example 4:

[0088] Compared with Example 1, this comparative example does not add EDC to the reaction raw materials, and the specific steps are as follows:

[0089]

[0090] In a reaction tube, 1 mmol of biotin was added, and under nitrogen protection, 146 mg of β-alanine and 101 mg of triethylamine were added at -78°C, and stirred at room temperature for 0.5 h; under nitrogen protection, 135 mg of HOBt and 2 mL of DCM were added at -78°C, and then stirred at 35°C for 24 h, the solvent was drained, washed with n-hexane, extracted with dichloromethane, the solvent was spin-dried, and silica gel column chromatography (the eluent was a mixture of petroleum ether and ethyl acetate in a volume ratio of 1:1) was performed to obtain 0 mg of the product, with a yield of 0%. This shows that without the participation of EDC, biotin and amino acids cannot react, and no amino acid derivatives can be obtained.

[0091] Comparative Example 5:

[0092] Compared with Example 1, this comparative example does not add HOBt to the reaction raw materials, and the specific steps are as follows:

[0093]

[0094] In a reaction tube, 1 mmol of biotin was added, and under nitrogen protection, 146 mg of β-alanine and 101 mg of triethylamine were added at -78°C, and stirred at room temperature for 0.5 h; under nitrogen protection, 157 mg of EDC and 2 mL of DCM were added at -78°C, and stirred at 35°C for 24 h, the solvent was drained, washed with n-hexane, extracted with dichloromethane, and the solvent was spin-dried. After silica gel column chromatography (the eluent was a mixture of petroleum ether and ethyl acetate in a volume ratio of 1:1), 0 mg of the product was obtained, and the yield was 0%. This shows that without the participation of HOBt, biotin and amino acids cannot react, and no amino acid derivatives can be obtained.

Claims

1. A method for preparing a biotinylated amino acid derivative, characterized in that: Amino acids and bases are added to biotin D-biotin, and then a catalyst and a solvent are added, the reaction is stirred, the solvent is drained, extraction is performed, and purification is performed to obtain a biotinylated amino acid derivative; the catalyst is 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide and 1-hydroxybenzotriazole.

2. The method for preparing the biotinylated amino acid derivative according to claim 1, characterized in that: The amino acid is at least one of glycine, alanine, valine, leucine, isoleucine, methionine, proline, tryptophan, serine, tyrosine, cysteine, phenylalanine, asparagine, glutamine, threonine, aspartic acid, glutamic acid, lysine, arginine, histidine, selenocysteine ​​or pyrrolysine.

3. The method for preparing the biotinylated amino acid derivative according to claim 1, characterized in that: The stirring reaction time is 1-48h; the temperature is 0°C-100°C.

4. The method for preparing the biotinylated amino acid derivative according to claim 1, characterized in that: The molar ratio of biotin, alkali, EDC, HOBt and amino acid is 1:(1-10):(1-10):(1-10):(1-10).

5. The method for preparing the biotinylated amino acid derivative according to claim 1, characterized in that: Under inert gas protection, amino acid and base were added at -78°C.

6. The method for preparing the biotinylated amino acid derivative according to claim 1, characterized in that: Under inert gas protection, the catalyst and solvent were added at -78°C.

7. The method for preparing the biotinylated amino acid derivative according to claim 1, characterized in that: The solvent is a polar solvent or a non-polar solvent, and is selected from at least one of ethanol, methanol, ethyl acetate, ether, dioxane, tetrahydrofuran, ethylene glycol dimethyl ether, 1,2-dichloroethane, dichloromethane, acetonitrile, 1,4-dioxane, benzene, toluene, xylene, trifluorotoluene, N,N-dimethylformamide, N,N-dimethylacetamide, and dimethyl sulfoxide.

8. The method for preparing the biotinylated amino acid derivative according to claim 1, characterized in that: The base is an inorganic base or an organic base. The inorganic base is lithium hydroxide, sodium hydroxide, potassium hydroxide, sodium carbonate, sodium bicarbonate, potassium carbonate, potassium bicarbonate, cesium carbonate, calcium carbonate, sodium phosphate, disodium hydrogen phosphate or sodium dihydrogen phosphate. The organic base is sodium methoxide, potassium ethoxide, potassium tert-butoxide, butyl lithium, phenyl lithium, lithium diisopropylamide, lithium hexamethyldisilazide, triethylamine or trimethylamine.

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