Preparation method of luminescent substrate

Abstract

The invention discloses a preparation method of a luminescent substrate. The luminescent substrate is obtained by reacting a compound shown in a general formula I with a compound shown in a general formula II under the catalysis of cuprous iodide, in the # imgabs0 #, R1 is a methyl group, a phenyl group or a benzyl group; r2 is methyl, ethyl, phenyl or benzyl; and R3 is chlorine or bromine. By using the preparation method of the luminescent substrate disclosed by the invention, the PS-atto chemiluminescent substrate can be prepared under milder conditions, and meanwhile, the yield of the PS-atto chemiluminescent substrate is improved.

Description

Technical Field

[0001] The present application relates to the field of chemiluminescent immunoassay technology, and in particular to a method for preparing a luminescent substrate. Background Art

[0002] Chemiluminescent immunoassay is an analytical method that determines the content of the corresponding compound by measuring the total amount of chemiluminescent light at a specific moment. It is also one of the more mature immunoassay methods, with the advantages of high specificity, high sensitivity, wide applicability, simple equipment, wide detection range and high degree of automation. According to the different luminescent agents in the chemiluminescent system, chemiluminescent immunoassay can be divided into direct chemiluminescent immunoassay, chemiluminescent enzyme immunoassay and electrochemiluminescent immunoassay.

[0003] Lumi-Phos HRP (PS-atto) is a special chemiluminescent substrate used in chemiluminescent enzyme immunoassay to detect the concentration of horseradish peroxidase (HRP) marker. In the prior art, the alkoxy sulfonate derivatization of PS-atto is mainly achieved by reacting with sultone under the action of sodium hydride (NaH) to obtain the final product. Since sodium hydride (NaH) is required in this method, the reaction conditions are relatively harsh, the post-processing is difficult and the yield is low.

[0004] Therefore, there is an urgent need to provide a new preparation method to prepare PS-atto chemiluminescent substrates under milder conditions and at the same time improve their yield. Summary of the invention

[0005] In order to solve the above-mentioned technical problems, the present application provides a method for preparing a luminescent substrate, wherein a compound of general formula I and a compound of general formula II are reacted under the catalysis of cuprous iodide to obtain;

[0006]

[0007] Wherein, R1 is methyl, phenyl or benzyl; R2 is methyl, ethyl, phenyl or benzyl; R3 is chlorine or bromine.

[0008] Preferably, the compound of general formula I and the compound of general formula II are obtained by reacting in the presence of an alkaline substance, cuprous iodide and a solvent.

[0009] Preferably, the alkaline substance is selected from one or more of sodium tert-butoxide and potassium tert-butoxide.

[0010] Preferably, the solvent is selected from one or more of tetrahydrofuran, DMF or DMSO.

[0011] Preferably, the molar ratio of the compound of formula I to the compound of formula II is 1:(1-2).

[0012] Furthermore, the molar ratio of the compound of formula I to the compound of formula II can be 1:1, 1:1.1, 1:1.2, 1:1.3, 1:1.4, 1:1.5, 1:1.6, 1:1.7, 1:1.8, 1:1.9, 1:2, but is not limited thereto.

[0013] Preferably, the molar ratio of the compound of formula I to the alkaline substance is 1:(1-2).

[0014] Furthermore, the molar ratio of the compound of formula I to the alkaline substance can be 1:1, 1:1.1, 1:1.2, 1:1.3, 1:1.4, 1:1.5, 1:1.6, 1:1.7, 1:1.8, 1:1.9, 1:2, but is not limited thereto.

[0015] Preferably, the ratio of the compound of formula I to the solvent is 1 mol:(1-3)L.

[0016] Preferably, the molar ratio of the compound of formula I to cuprous iodide is 1:(0.01-0.1).

[0017] Furthermore, the molar ratio of the compound of formula I to cuprous iodide can be 1:0.01, 1:0.02, 1:0.03, 1:0.04, 1:0.05, 1:0.06, 1:0.07, 1:0.08, 1:0.09, 1:0.1, but is not limited thereto.

[0018] Preferably, the reaction is carried out at room temperature.

[0019] Preferably, the reaction time is 1 to 5 hours.

[0020] Furthermore, the reaction time is 1h, 1.5h, 2h, 2.5h, 3h, 3.5h, 4h, 4.5h, 5h, but is not limited thereto.

[0021] In summary, this application has the following beneficial effects:

[0022] 1. The preparation method provided in the present application is carried out at room temperature under simple reaction conditions, and does not use harsh reaction conditions such as sodium hydride (NaH), and is simple and easy to obtain.

[0023] 2. The post-treatment method of the preparation method provided in the present application is very simple, and the target product can be obtained by only washing with saturated brine, followed by separation and recrystallization. DETAILED DESCRIPTION

[0024] Example

[0025] The following examples are methods for preparing the luminescent substrate of the present application.

[0026] Example 1

[0027] 20 mL THF, 0.34 g raw material 1, 0.14 g sodium tert-butoxide and 0.01 g CuI were added to a three-necked flask and stirred, and 0.24 g raw material 2 was added dropwise, and the reaction was carried out at room temperature for 1 hour. After the reaction was completed, the reaction solution was filtered, washed with 4 mL saturated brine, and then separated. The organic phase was concentrated and dried, and recrystallized from ethanol to obtain 0.28 g of the product. The product structure is shown below, and the yield is 57%.

[0028] Raw material 1 is:

[0029]

[0030] Raw material 2 is:

[0031]

[0032] The products are:

[0033]

[0034] Example 2

[0035] 10 mL THF, 0.36 g raw material 1, 0.22 g potassium tert-butoxide, and 0.02 g CuI were added to a three-necked flask and stirred, and 0.32 g raw material 2 was added dropwise, and the reaction was carried out at room temperature for 2 h. After the reaction was complete, the reaction solution was filtered, washed with 2 mL saturated brine, and then separated. The organic phase was concentrated and dried, and recrystallized from ethanol to obtain 0.31 g of the product. The product structure is shown below, and the yield is 59%.

[0036] Raw material 1 is:

[0037]

[0038] Raw material 2 is:

[0039]

[0040] The products are:

[0041]

[0042] Example 3

[0043] 10 mL DMF, 0.48 g raw material 1, 0.1 g sodium tert-butoxide, and 0.002 g CuI were added to a three-necked flask and stirred, and 0.16 g raw material 2 was added dropwise, and the reaction was carried out at room temperature for 5 h. After the reaction was complete, the reaction solution was filtered, washed with 2 mL saturated brine, and then separated. The organic phase was concentrated and dried, and recrystallized from ethanol to obtain 0.31 g of the product. The product structure is shown below, and the yield is 49%.

[0044] Raw material 1 is:

[0045]

[0046] Raw material 2 is:

[0047]

[0048] The products are:

[0049]

[0050] Example 4

[0051] 30 mL THF, 0.48 g raw material 1, 0.1 g sodium tert-butoxide, and 0.01 g CuI were added to a three-necked flask and stirred, and 0.20 g raw material 2 was added dropwise, and the reaction was carried out at room temperature for 4 h. After the reaction was complete, the reaction solution was filtered, washed with 6 mL saturated brine, and then separated. The organic phase was concentrated and dried, and recrystallized from ethanol to obtain 0.30 g of the product. The product structure is shown below, and the yield is 47%.

[0052] Raw material 1 is:

[0053]

[0054] Raw material 2 is:

[0055]

[0056] The products are:

[0057]

[0058] Example 5

[0059] 10 mL DMSO, 0.43 g raw material 1, 0.14 g sodium tert-butoxide, and 0.01 g CuI were added to a three-necked flask and stirred, and 1.5 mmol 0.3 g raw material 2 was added dropwise, and the reaction was carried out at room temperature for 3 h. After the reaction was complete, the reaction solution was filtered, washed with 2 mL saturated brine, and then separated. The organic phase was concentrated and dried, and recrystallized from ethanol to obtain 0.27 g of the product. The product structure is shown below, and the yield is 47%.

[0060] Raw material 1 is:

[0061]

[0062] Raw material 2 is:

[0063]

[0064] The products are:

[0065]

[0066] Example 6

[0067] 30 mL THF, 0.34 g raw material 1, 0.29 g sodium tert-butoxide, and 0.04 g CuI were added to a three-necked flask and stirred, and 0.48 g raw material 2 was added dropwise, and the reaction was carried out at room temperature for 1 h. After the reaction was complete, the reaction solution was filtered, washed with 6 mL saturated brine, and then separated. The organic phase was concentrated and dried, and recrystallized from ethanol to obtain 0.20 g of the product. The product structure is shown below, and the yield is 45%.

[0068] Raw material 1 is:

[0069]

[0070] Raw material 2 is:

[0071]

[0072] The products are:

[0073]

[0074] Example 7

[0075] 10 mL THF, 0.34 g raw material 1, 0.05 g sodium tert-butoxide, and 0.002 g CuI were added to a three-necked flask and stirred, and 0.08 g raw material 2 was added dropwise, and the reaction was carried out at room temperature for 1 h. After the reaction was complete, the reaction solution was filtered, washed with 2 mL saturated brine, and then separated. The organic phase was concentrated and dried, and recrystallized from ethanol to obtain 0.12 g of the product. The product structure is shown below, and the yield is 24%.

[0076] Raw material 1 is:

[0077]

[0078] Raw material 2 is:

[0079]

[0080] The products are:

[0081]

[0082] Example 8

[0083] 50 mL DMSO, 0.34 g raw material 1, 0.14 g sodium tert-butoxide, and 0.03 g CuI were added to a three-necked flask and stirred, and 0.24 g raw material 2 was added dropwise, and the reaction was carried out at room temperature for 1 h. After the reaction was complete, the reaction solution was filtered, washed with 6 mL saturated brine, and then separated. The organic phase was concentrated and dried, and recrystallized from ethanol to obtain 0.24 g of the product. The product structure is shown below, and the yield is 50%.

[0084] Raw material 1 is:

[0085]

[0086] Raw material 2 is:

[0087]

[0088] The products are:

[0089]

[0090] Example 9

[0091] 20 mL THF, 0.34 g raw material 1, 0.14 g sodium tert-butoxide, and 0.01 g CuI were added to a three-necked flask and stirred, and 0.24 g raw material 2 was added dropwise, and the mixture was reacted at 60°C for 1 h. After the reaction was complete, the reaction solution was filtered, washed with 4 mL saturated brine, and then separated. The organic phase was concentrated and dried, and recrystallized from ethanol to obtain 0.19 g of the product. The product structure is shown below, and the yield is 40%.

[0092] Raw material 1 is:

[0093]

[0094] Raw material 2 is:

[0095]

[0096] The products are:

[0097]

[0098] Example 10

[0099] 20 mL THF, 0.34 g raw material 1, 0.14 g sodium tert-butoxide, and 0.01 g CuI were added to a three-necked flask and stirred, and 0.24 g raw material 2 was added dropwise, and the mixture was reacted at 10°C for 1 h. After the reaction was complete, the reaction solution was filtered, washed with 4 mL saturated brine, and then separated. The organic phase was concentrated and dried, and recrystallized from ethanol to obtain 0.20 g of the product. The product structure is shown below, and the yield is 45%.

[0100] Raw material 1 is:

[0101]

[0102] Raw material 2 is:

[0103]

[0104] The products are:

[0105]

[0106] Embodiment 11

[0107] 20 mL THF, 0.34 g raw material 1, 0.14 g sodium tert-butoxide, and 0.01 g CuI were added to a three-necked flask and stirred, and 0.24 g raw material 2 was added dropwise, and the reaction was carried out at room temperature for 0.5 h. After the reaction was complete, the reaction solution was filtered, washed with 4 mL saturated brine, and then separated. The organic phase was concentrated and dried, and recrystallized from ethanol to obtain 0.23 g of the product. The product structure is shown below, and the yield is 47%.

[0108] Raw material 1 is:

[0109]

[0110] Raw material 2 is:

[0111]

[0112] The products are:

[0113]

[0114] Example 12

[0115] 20 mL THF, 0.34 g raw material 1, 0.14 g sodium tert-butoxide, and 0.01 g CuI were added to a three-necked flask and stirred, and 0.24 g raw material 2 was added dropwise, and the reaction was carried out at room temperature for 8 h. After the reaction was complete, the reaction solution was filtered, washed with 4 mL saturated brine, and then separated. The organic phase was concentrated and dried, and recrystallized from ethanol to obtain 0.27 g of the product. The product structure is shown below, and the yield is 55%.

[0116] Raw material 1 is:

[0117]

[0118] Raw material 2 is:

[0119]

[0120] The products are:

[0121]

[0122] Embodiment 13

[0123] 10 mL THF and 10 mL DMSO, 0.34 g raw material 1, 0.14 g sodium tert-butoxide, and 0.01 g CuI were added to a three-necked flask and stirred, and 0.24 g raw material 2 was added dropwise, and the reaction was carried out at room temperature for 1 hour. After the reaction was complete, the reaction solution was filtered, washed with 4 mL saturated brine, and then separated. The organic phase was concentrated and dried, and recrystallized from ethanol to obtain 0.29 g of the product. The product structure is shown below, and the yield is 59%.

[0124] Raw material 1 is:

[0125]

[0126] Raw material 2 is:

[0127]

[0128] The products are:

[0129]

[0130] It can be seen from the above examples that the preparation method disclosed in the present application has relatively simple reaction conditions, does not use sodium hydride (NaH) used in the prior art, the reaction can be carried out at room temperature, the conditions are relatively mild, and has a relatively high yield compared to the prior art.

[0131] In addition, the present application uses a very simple post-processing method to process the reaction products and obtain the target product. Compared with the complicated post-processing process of the prior art, the preparation method provided by the present application is easier to carry out industrial production, which greatly reduces the production cost.

[0132] This specific embodiment is merely an explanation of the present application and is not a limitation of the present application. After reading this specification, those skilled in the art may make modifications to the present embodiment without any creative contribution as needed, but such modifications are protected by the patent law as long as they are within the scope of the claims of the present application.

Claims

1. A method for preparing a luminescent substrate, characterized in that: The compound of general formula I and the compound of general formula II are reacted under the catalysis of cuprous iodide to obtain; Wherein, R1 is methyl, phenyl or benzyl; R2 is methyl, ethyl, phenyl or benzyl; R3 is chlorine or bromine.

2. The preparation method according to claim 1, characterized in that: The compound of general formula I and the compound of general formula II are reacted in the presence of an alkaline substance, cuprous iodide and a solvent to obtain the product.

3. The preparation method according to claim 1, characterized in that: The alkaline substance is selected from one or more of sodium tert-butoxide and potassium tert-butoxide.

4. The preparation method according to claim 1, characterized in that: The solvent is selected from one or more of tetrahydrofuran, DMF or DMSO.

5. The preparation method according to any one of claims 1 to 4, characterized in that: The molar ratio of the compound of general formula I to the compound of general formula II is 1:(1-2).

6. The preparation method according to any one of claims 1 to 4, characterized in that: The molar ratio of the compound of the general formula I to the alkaline substance is 1:(1-2).

7. The preparation method according to any one of claims 1 to 4, characterized in that: The ratio of the compound of formula I to the solvent is 1 mol:(1-3)L.

8. The preparation method according to any one of claims 1 to 4, characterized in that: The molar ratio of the compound of the general formula I to cuprous iodide is 1:(0.01-0.1).

9. The preparation method according to claim 1, characterized in that: The reaction was carried out at room temperature.

10. The preparation method according to claim 1, characterized in that: The reaction time is 1 to 5 hours.

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