CN104459099B - Derivatives of porphyrin detects the application in inflammation related disease reagent in preparation - Google Patents

Abstract

The invention discloses the application of a porphyrin derivative in the preparation of a reagent for detecting inflammation-related diseases. The porphyrin derivative can be tetra-p-sulfonic acid phenyl porphyrin. The inflammation-related disease is rheumatoid arthritis. Porphyrin derivatives have stable and good optical and electrochemical properties and excellent biocompatibility, and metabolites in body fluids or urine of related diseases can significantly change the optical properties of porphyrin derivatives (including ultraviolet-visible absorption spectra and fluorescence spectra, etc.) and electrochemical properties. Utilizing this characteristic, the present invention proposes for the first time the application of porphyrin derivatives in the preparation of reagents for detecting inflammation-related diseases, and expands the new application of porphyrin derivatives.

Description

Application of porphyrin derivatives in the preparation of reagents for detecting inflammation-related diseases

technical field

The invention relates to the field of biochemical pharmacy, in particular to the application of porphyrin derivatives in the preparation of reagents for detecting inflammation-related diseases (such as rheumatoid arthritis).

Background technique

As we all know, inflammation is a very common and important basic pathological process. Trauma infection on the body surface and most common and frequently-occurring diseases of various organs (such as boils, carbuncles, pneumonia, hepatitis, nephritis, etc.) are all inflammatory diseases. The defensive response of living tissue with vascular system to injury factors, and correspondingly produces many kinds of metabolites in this process. Rheumatoid arthritis is one of the chronic systemic autoimmune diseases mainly characterized by joint disease. The main clinical manifestations are joint swelling and pain caused by the synovial membrane of the facet joints, followed by cartilage destruction and joint space narrowing. In the late stage, severe bone destruction and absorption lead to joint stiffness, deformity, and dysfunction. The disease is mostly a recurrent disease, with a high disability rate and poor prognosis, and there is no good cure at present. Controlling rheumatoid drugs can produce long-term relief, delay or stop the progression of the disease. Anti-inflammatory and analgesic drugs can relieve pain and improve stiffness, but they cannot prevent joint damage or slow down the progression of the disease. In most cases, when the symptoms are obvious, it is already in the middle and late stage. Therefore, finding an early detection method may be able to better cure this kind of disease, and it is also the direction of early prevention in clinical medicine in the future.

Porphyrin derivatives are usually divided into fat-soluble porphyrin compounds and water-soluble porphyrin compounds. This substance has certain photosensitive properties and can effectively release singlet oxygen under the action of ultraviolet light or visible light. Early porphyrins were obtained from natural products containing porphyrin compounds, such as heme and chlorophyll, through extraction, separation, and purification. Currently, there are two ways to obtain target porphyrin molecules: structural modification of natural porphyrins and total synthesis of porphyrin compounds. Although the structural modification of natural porphyrins can be easily modified, it is limited by the structure itself, and the choice of functional groups on the outer ring is also very limited. In addition, the physiological activity of porphyrin compounds is also limited. Therefore, people generally need to obtain porphyrin molecules with specific physiological activities and functions through total synthesis. The tetra-p-sulfonic acid phenyl porphyrin (TSPP) used in the present invention is a kind of water-soluble sulfonic acid based porphyrin derivative, which has stable fluorescence characteristics and good biocompatibility.

Contents of the invention

The technical problem to be solved by the present invention is to provide a change in the optical and electrochemical properties of porphyrin derivatives and inflammation-related metabolites to achieve early rapid and highly sensitive detection of rheumatoid arthritis, which has broad application value and outlook.

In order to solve the above-mentioned technical problems, the technical scheme adopted in the present invention is to provide a kind of application of tetra-p-sulfonic acid phenylporphyrin in the preparation and detection of rheumatoid arthritis reagent, which is characterized in that, using four-p- -The porphyrin ring structure of sulfophenylporphyrin and the sulfonic acid group bind and coordinate with specific proteins in inflammatory metabolites, thereby changing the optical and optical properties of tetra-p-sulfophenylporphyrin Principles of electrochemical properties, preparation of reagents for the detection of rheumatoid arthritis.

The beneficial effects of the present invention are: compared with the prior art, the present invention has the following advantages: because porphyrin derivatives have stable and good optical and electrochemical properties and excellent biocompatibility, and the body fluid or urine of related diseases The metabolites in the porphyrin derivatives can significantly change the optical properties (including UV-visible absorption spectra and fluorescence spectra, etc.) and electrochemical properties. Utilizing this characteristic, the present invention proposes for the first time the application of porphyrin derivatives in the preparation of reagents for detecting inflammation-related diseases, and expands the new application of porphyrin derivatives.

Description of drawings

Figure 1 is the UV-Vis absorption spectrum of TSPP with a concentration of 0.01mg/ml

Fig. 2 is the comparison chart of the changes of the ultraviolet-visible absorption peaks between the control group and the experimental group in the experiment of using porphyrin derivatives described in the present invention——TSPP to detect rheumatoid arthritis

detailed description

The present invention will be further described below in conjunction with the accompanying drawings of the description

Since tetra-p-sulfophenylporphyrin (TSPP) has a porphyrin ring structure and a sulfonic acid group similar to most porphyrin derivatives, and the detected proteins in related metabolites are easily Combining and coordinating with porphyrin ring structure and sulfonic acid group, thereby changing the optical and electrochemical properties of porphyrin derivatives, the following examples take tetra-p-sulfonic acid phenylporphyrin (TSPP) as an example for porphyrin The application of porphyrin derivatives in the preparation of reagents for detecting inflammation-related diseases will be described. Other porphyrin derivatives with similar structures can also be used in the preparation of reagents for detecting inflammation-related diseases.

In the following examples, the body fluid (such as blood) or urine of the normal experimental model group is taken from normal male DBA/1 mice or other normal control groups; the body fluid or urine of the inflammation-related disease experimental model group is taken from collagen-induced Early CIA male DBA/1 mice or other experimental model groups of inflammation-related diseases.

Below in conjunction with specific embodiment, further illustrate the present invention.

Example 1: Using TSPP optical properties to detect inflammation-related metabolites such as rheumatoid arthritis, the specific steps are as follows:

1 Utilize the UV-visible spectrophotometer and the fluorescence spectrophotometer to characterize the optical (UV-visible absorption spectrum and fluorescence spectrum) characteristics of TSPP (0.01mg/ml), and obtain the UV-visible absorption peak A of the standard solution TSPP (0.01mg/ml) _TSPP (as shown in Figure 1 ) and the fluorescence emission peak I _TSPP obtained by excitation with 515nm excitation light.

2 Take 10 μl of body fluid or urine from the normal experimental model group and 10 μl of the body fluid or urine from the inflammation-related disease experimental model group, add TSPP (0.01 mg/ml) standard solution dropwise, and use a UV-visible spectrophotometer and a fluorescence spectrophotometer Characterize the optical (ultraviolet-visible absorption spectrum and fluorescence spectrum) properties of the mixed solution, obtain the ultraviolet-visible absorption peak value A _UN of each mixed solution and the fluorescence emission peak value I _UN obtained by exciting with 515nm excitation light, and calculate the standard solution TSPP (0.01 mg/ml) and the change values ΔA=A _TSPP -A _UN and ΔI=I _TSPP -I _UN between the normal experimental model group and the inflammation-related disease experimental model group.

3 The test results show that the peak change of the UV-visible absorption peak ΔA _NOR and the change of the peak value of the fluorescence emission ΔI _NOR in the normal experimental model group are much smaller than the change ΔA _ILL of the peak value of the ultraviolet-visible absorption peak and the change ΔI of the peak value of the fluorescence emission peak of the experimental model group of inflammation-related diseases _ill .

Example 2: Using the electrochemical characteristics of TSPP to detect inflammation-related metabolites such as rheumatoid arthritis, the specific steps are as follows:

1. Use the electrochemical workstation to characterize the electrochemical characteristics of TSPP standard solution (0.01mg/ml), and obtain the electrochemical differential pulse voltammetry curve (DPV _TSPP ) of the standard solution and several corresponding electrochemical peaks.

2 Take 10 μl of body fluid or urine from the normal experimental model group and 10 μl of the body fluid or urine from the inflammation-related disease experimental model group, add TSPP (0.01 mg/ml) standard solution dropwise, and use the electrochemical workstation to characterize the electrochemical reaction of the above mixed solution. The electrochemical differential pulse voltammetry curves (DPV _NOR and DPV _ILL ) of each mixed solution and corresponding electrochemical peaks were obtained.

3 The results showed that there was no significant difference between the DPV _NOR of the normal experimental model group and the DPV _TSPP of the standard solution, and the corresponding electrochemical peak position and peak value did not change significantly; while the DPV _ILL of the inflammation-related disease experimental model group was significantly different from the DPV _TSPP of the standard solution Differently, many corresponding electrochemical peaks disappeared or the peaks became smaller due to the addition of body fluid or urine from the experimental model group of inflammation-related diseases.

Show by above concrete example result: the porphyrin derivative that this method adopts has good optical characteristic and electrochemical characteristic, and the body fluid or urine of inflammatory disease model animal and patient can significantly change the optical characteristic of porphyrin derivative (including UV-Vis spectra and fluorescence spectra, etc.) and electrochemical properties. The method takes full advantage of the excellent properties of the porphyrin derivatives and their effects on body fluids and urine in inflammatory diseases, is simple, easy to implement, rapid and specific, and has broad medical clinical application value and prospects.

The basic principles and main features of the present invention and the advantages of the present invention have been shown and described above. Those skilled in the industry should understand that the present invention is not limited by the above-mentioned embodiments. What are described in the above-mentioned embodiments and the description only illustrate the principle of the present invention. Without departing from the spirit and scope of the present invention, the present invention will also have Variations and improvements are possible, which fall within the scope of the claimed invention. The protection scope of the present invention is defined by the appended claims and their equivalents.

Claims (1)

1. the application of four-p-sulfonic group phenyl porphyrin in preparation detection type rheumatic arthritis reagent, it is characterized in that, the porphyrin ring texture of four-p-sulfonic group phenyl porphyrin and sulfonic group is utilized to combine with the specific proteins in inflammation metabolin, coordination, thus change the four-p-optics of sulfonic group phenyl porphyrin and the principle of electrochemical properties, for the preparation of the reagent of detection type rheumatic arthritis.