CN112540143A

CN112540143A - Aryl sulfatase inhibitor and preparation method and application thereof

Info

Publication number: CN112540143A
Application number: CN201910900083.2A
Authority: CN
Inventors: 刘则华; 张俊; 钟姝姝; 党志
Original assignee: South China University of Technology SCUT
Current assignee: South China University of Technology SCUT
Priority date: 2019-09-23
Filing date: 2019-09-23
Publication date: 2021-03-23

Abstract

The invention belongs to the field of medicine, and discloses an arylsulfatase inhibitor, and a preparation method and application thereof. Adding dodecyl sulfate into a solvent to obtain a solution, and preparing the aryl sulfatase inhibitor. Compared with the conventional medical treatment inhibitor STX64, the invention has the advantages of low effective inhibition concentration, capability of inhibiting in different degrees according to requirements, simple operation, low cost, high speed and the like, and has wide application prospect in the fields of environment and medicine.

Description

Aryl sulfatase inhibitor and preparation method and application thereof

Technical Field

The invention belongs to the field of medicine, and particularly relates to an arylsulfatase inhibitor, and a preparation method and application thereof.

Background

Arylsulfatase (arylsulfatase) is widely present in various organs of humans and animals, and also in various environmental media such as domestic sewage. The important function of the aryl sulfatase is to convert sulfate conjugates such as natural estrogen into natural estrogen, i.e. desulfate conjugates. In animals, arylsulfatase may be closely related to the normal physiological functions of the human body. For example, some patients with breast cancer use drugs to inhibit arylsulfatase, thereby achieving the goal of controlling the progression of breast cancer. The current method for effectively inhibiting the arylsulfatase uses an STX64 inhibitor, but the method is mainly used in the medical field and has the defects of high price, short drug effect, side effect and the like.

Disclosure of Invention

In view of the above disadvantages, the primary object of the present invention is to provide an arylsulfatase inhibitor.

Another object of the present invention is to provide a process for producing the above arylsulfatase inhibitor.

It is a further object of the present invention to provide the use of the above arylsulfatase inhibitors.

The invention is realized by the following technical scheme:

a preparation method of an arylsulfatase inhibitor comprises the following steps: adding dodecyl sulfate into a solvent to obtain a solution, and preparing the aryl sulfatase inhibitor.

Preferably, the concentration of the dodecyl sulfate in the solution is 120 to 300 mu mol/L.

Preferably, the lauryl sulfate is one or more of sodium lauryl sulfate, potassium lauryl sulfate and calcium lauryl sulfate, and more preferably sodium lauryl sulfate.

Preferably, the solvent is one or two of physiological saline and Tris-HCl buffer solution.

Preferably, the pH value of the Tris-HCl buffer solution is 5-7, and more preferably 5.8.

The aryl sulfatase inhibitor prepared by the preparation method of the aryl sulfatase inhibitor.

The application of the arylsulfatase inhibitor in preparing a breast cancer medicament.

Compared with the prior art, the invention has the following advantages and beneficial effects:

(1) the greatest advantage of the present invention is the economy compared to thousands of dollars required for a few milligrams of conventional inhibitors such as STX 64.

(2) The 50% inhibitory concentration in the present invention is 52mg/L (equivalent to 181. mu. mol/L) compared to the 50% inhibitory concentration of conventional inhibitors such as STX64, which requires several tens of milligrams.

(3) The invention has the advantages of simple operation, good effect, low price and the like, can carry out inhibition of different degrees according to the needs, and has wide application prospect in the aspect of medical treatment.

Detailed Description

The present invention will be described in further detail with reference to examples, but the embodiments of the present invention are not limited thereto.

Examples arylsulfatase was purchased from Sigma Aldrich (model EC 3.1.6.1, from snail extracts); p-nitrophenol sulfuric acid conjugate (purity > 98%) was purchased from Sigma Aldrich.

Example 1

3mL of Tris-HCl buffer (pH 5.8) was put in a glass tube, 5.22U of arylsulfatase was added to the glass tube, 1mol/L sodium dodecyl sulfate solution was added to the Tris-HCl buffer solution so that the concentration of sodium dodecyl sulfate was 100. mu. mol/L, 800. mu. mol/L of the combined p-nitrophenol and sulfuric acid was added thereto, the glass tube was placed in a 37 ℃ water bath and reacted for 1 hour, then the reaction was terminated with 2mL of 0.5mol/L sodium hydroxide, and the activity of arylsulfatase was measured by a standard method (reference to Tabatabai and Brenner, Soil Science of America Journal,1970,34: 225-229).

Example 2

3mL of Tris-HCl buffer (pH 5.8) was placed in a glass tube, 5.22U of arylsulfatase was added to the glass tube, 1mol/L sodium dodecyl sulfate solution was added to the Tris-HCl buffer solution so that the concentration of sodium dodecyl sulfate was 120. mu. mol/L, 800. mu. mol/L of the combined p-nitrophenol and sulfuric acid was added thereto, the glass tube was placed in a 37 ℃ water bath and reacted for 1 hour, then the reaction was terminated with 2mL of 0.5mol/L sodium hydroxide, and the activity of arylsulfatase was measured by a standard method (reference to Tabatabai and Brenner, Soil Science of America Journal,1970,34: 225-229).

Example 3

3mL of Tris-HCl buffer (pH 5.8) was placed in a glass tube, 5.22U of arylsulfatase was added to the glass tube, 1mol/L sodium dodecyl sulfate solution was added to the Tris-HCl buffer solution to make the concentration of sodium dodecyl sulfate 140. mu. mol/L, 800. mu. mol/L of the combined p-nitrophenol and sulfuric acid was added thereto, the glass tube was placed in a 37 ℃ water bath and reacted for 1 hour, then the reaction was terminated with 2mL of 0.5mol/L sodium hydroxide, and the arylsulfatase activity was measured by a standard method (reference to Tabatabai and Brenner, Soil Science of America Journal,1970,34: 225-229).

Example 4

3mL of Tris-HCl buffer (pH 5.8) was placed in a glass tube, 5.22U of arylsulfatase was added to the glass tube, 1mol/L sodium dodecyl sulfate solution was added to the Tris-HCl buffer solution to give a concentration of 160. mu. mol/L sodium dodecyl sulfate, 800. mu. mol/L of the combined p-nitrophenol-sulfuric acid was added thereto, the glass tube was placed in a 37 ℃ water bath and reacted for 1 hour, 2mL of 0.5mol/L sodium hydroxide was used to terminate the reaction, and the arylsulfatase activity was measured by a standard method (Tabatabai and Brenner, Soil Science Society of America Journal,1970,34: 225-229).

Example 5

3mL of Tris-HCl buffer (pH 5.8) was put in a glass tube, 5.22U of arylsulfatase was added to the glass tube, 1mol/L of a sodium dodecyl sulfate solution was added to the Tris-HCl buffer so that the concentration of sodium dodecyl sulfate in the Tris-HCl buffer was 200. mu. mol/L, a p-nitrophenol sulfuric acid conjugate was added in accordance with the standard of 800. mu. mol/L, the glass tube was placed in a 37 ℃ water bath and reacted for 1 hour, then the reaction was terminated with 2mL of 0.5mol/L of sodium hydroxide, and the arylsulfatase activity was measured by a standard method (reference to Tabatabai and Brenner, Soil Society of America Journal,1970,34: 225-.

Example 6

3mL of Tris-HCl buffer (pH 5.8) was placed in a glass tube, 5.22U of arylsulfatase was added to the glass tube, 1mol/L sodium dodecyl sulfate solution was added to the Tris-HCl buffer solution so that the concentration of sodium dodecyl sulfate was 220. mu. mol/L, 800. mu. mol/L of the combined p-nitrophenol and sulfuric acid was added thereto, the glass tube was placed in a 37 ℃ water bath and reacted for 1 hour, then the reaction was terminated with 2mL of 0.5mol/L sodium hydroxide, and the activity of arylsulfatase was measured by a standard method (reference to Tabatabai and Brenner, Soil Science of America Journal,1970,34: 225-229).

Example 7

3mL of Tris-HCl buffer (pH 5.8) was placed in a glass tube, 5.22U of arylsulfatase was added to the glass tube, 1mol/L sodium dodecyl sulfate solution was added to the Tris-HCl buffer solution so that the concentration of sodium dodecyl sulfate was 240. mu. mol/L, 800. mu. mol/L of the p-nitrophenol-sulfuric acid conjugate was added, the glass tube was placed in a 37 ℃ water bath and reacted for 1 hour, 2mL of 0.5mol/L sodium hydroxide was used to terminate the reaction, and the arylsulfatase activity was measured by a standard method (Tabatabai and Brenner, Soil Science Society of America Journal,1970,34: 225-.

Example 8

3mL of Tris-HCl buffer (pH 5.8) was placed in a glass tube, 5.22U of arylsulfatase was added to the glass tube, 1mol/L sodium dodecyl sulfate solution was added to the Tris-HCl buffer solution to make the concentration of sodium dodecyl sulfate 260. mu. mol/L, 800. mu. mol/L of the p-nitrophenol-sulfuric acid conjugate was added, the glass tube was placed in a 37 ℃ water bath and reacted for 1 hour, 2mL of 0.5mol/L sodium hydroxide was used to terminate the reaction, and the arylsulfatase activity was measured by a standard method (Tabatabai and Brenner, Soil Science Society of America Journal,1970,34: 225-229).

Example 9

3mL of Tris-HCl buffer (pH 5.8) was put in a glass tube, 5.22U of arylsulfatase was added to the glass tube, 1mol/L of a sodium dodecyl sulfate solution was added to the Tris-HCl buffer so that the concentration of sodium dodecyl sulfate in the Tris-HCl buffer was 300. mu. mol/L, a p-nitrophenol sulfuric acid conjugate was added in accordance with 800. mu. mol/L, the glass tube was placed in a 37 ℃ water bath and reacted for 1 hour, then the reaction was terminated with 2mL of 0.5mol/L of sodium hydroxide, and the arylsulfatase activity was measured by a standard method (reference to Tabatabai and Brenner, Soil Science of America Journal,1970,34: 225-229).

Comparative example

3mL of Tris-HCl buffer (pH 5.8) was put in a glass tube, 5.22U of arylsulfatase was added to the glass tube, and then a p-nitrophenol-sulfuric acid conjugate was added thereto in an amount of 800. mu. mol/L, the glass tube was placed in a water bath at 37 ℃ for 1 hour, and then the reaction was terminated with 2mL of 0.5mol/L sodium hydroxide, and the arylsulfatase activity was measured by a standard method (refer to Tabatabai and Brerner, Soil Science of America Journal,1970,34: 225-.

The test results of examples 1 to 9 and comparative example are shown in Table 1.

TABLE 1 summary of test results

1U is defined as producing 1. mu.g/ml p-nitrophenol per hour

As can be seen from Table 1: when the concentration of sodium lauryl sulfate was 0. mu. mol/L, that is, when it was a comparative example, it had no inhibitory effect on arylsulfatase. Meanwhile, the inhibitory effect of the sodium dodecyl sulfate on arylsulfatase is stronger and stronger as the concentration of the sodium dodecyl sulfate is increased.

Tris-HCl buffer (pH 5.8) in the examples of the present invention was replaced with physiological saline (pH 5.8), which also had a good inhibitory effect on arylsulfatase.

The lauryl sodium sulfate in the embodiment of the invention is replaced by the lauryl potassium sulfate and the lauryl calcium sulfate, and the lauryl potassium sulfate and the lauryl calcium sulfate also have good inhibition effect on aryl sulfatase.

Without being limited to the above embodiments, other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the invention should be construed as equivalents thereof and are intended to be included in the scope of the invention.

Claims

1. A preparation method of an arylsulfatase inhibitor is characterized by comprising the following steps: adding dodecyl sulfate into a solvent to obtain a solution, and preparing the aryl sulfatase inhibitor.

2. The method of claim 1, wherein the concentration of dodecyl sulfate in the solution is 120 μmol/L to 300 μmol/L.

3. The method of claim 1, wherein the solvent is one or both of physiological saline and Tris-HCl buffer.

4. The method of claim 1, wherein the lauryl sulfate is one or more of sodium lauryl sulfate, potassium lauryl sulfate, and calcium lauryl sulfate.

5. The method of claim 3, wherein the Tris-HCl buffer has a pH of 5 to 7.

6. The method of claim 5, wherein the Tris-HCl buffer has a pH of 5.8.

7. The method of claim 4, wherein the lauryl sulfate is sodium lauryl sulfate.

8. An arylsulfatase inhibitor produced by the process for producing an arylsulfatase inhibitor according to any one of claims 1 to 7.

9. Use of an arylsulfatase inhibitor according to claim 8 for the preparation of a medicament for treating breast cancer.