CN117696017B - A blood purification adsorption modified material and preparation method thereof - Google Patents

Abstract

The invention belongs to the field of blood purification materials, and particularly relates to a blood purification adsorption modified material and a preparation method thereof. The preparation method comprises the following steps: soaking the blood purification and adsorption material in an oxidant acid solution, and performing reaction and cleaning to obtain a carboxylation purification and adsorption material; EDC/NHS-MES buffer solution is firstly added to the surface of the carboxylation purification adsorption material for reaction, then amine-containing hydrophobic compound-ethanol/water solution is added for reaction, finally NO catalyst solution is added for reaction, and after completion, the blood purification adsorption modified material is obtained through cleaning and drying. According to the invention, the hydrophobic compound and the NO catalyst are modified on the surface of the blood purification adsorption material, so that the biocompatibility of the adsorption material can be effectively ensured, and the coagulation and inflammatory reaction can be reduced; also improves the removal of non-polar harmful substances, fat-soluble drugs, fat-soluble substances, non-polar bacterial endotoxins and harmful proteins which can coordinate with NO catalysts.

Description

A blood purification adsorption modified material and preparation method thereof

Technical Field

The invention belongs to the field of blood purification materials, and in particular relates to a blood purification adsorption modified material and a preparation method thereof.

Background technique

Blood purification therapy is an extracorporeal circulation life support technology that has been used clinically to treat various diseases, such as poisoning (drugs, pesticides, heavy metals, biological toxins, drugs, etc.), uremia, liver disease, hyperlipidemia, sepsis, autoimmune diseases, etc. Patients with these diseases accumulate a large amount of exogenous or endogenous toxins in their bodies due to the ingestion of toxic substances or dysfunction of their own detoxification, immune, and metabolic systems. In the case of failure of the patient's own detoxification organs (kidneys, liver) function, blood purification therapy can lead the patient's blood out of the body and pass it through a purification device to purify and remove toxins in the blood, and then return the purified blood to the patient's body, thereby replacing the patient's detoxification system to achieve toxin removal. The methods of blood purification include: hemodialysis, hemofiltration and hemoperfusion. Among them, the core purification material of hemoperfusion is hemoperfusion adsorbent. Based on the principle of "adsorption", toxins in the blood are removed through the broad-spectrum adsorption and specific adsorption of the adsorbent. Broad-spectrum adsorbents play an important role in this process. They can simultaneously remove a variety of harmful substances, including but not limited to urea, creatinine, bilirubin, endotoxins, inflammatory factors, drug residues, toxins, etc., shortening the treatment time and frequency, reducing the burden on organs, and thus reducing patients' pain and financial pressure.

Although broad-spectrum adsorption columns have been widely used in clinical practice, they still face many difficulties and challenges during use. These include: 1) Balance between selectivity and affinity: Broad-spectrum adsorbents need to have high adsorption capacity for a variety of substances, but at the same time do not want to produce excessive adsorption on normal blood components (such as proteins, electrolytes, nutrients, etc.). Therefore, it is a major challenge to design and prepare broad-spectrum adsorbents with both high selectivity and high affinity. 2) Biocompatibility: The adsorbent needs to be in direct contact with the blood, so it must have good biocompatibility to avoid inducing adverse reactions such as thrombosis, inflammation or immune response. At present, many adsorbents have not yet achieved ideal biocompatibility. 3) Easy to clog: Blood contains a variety of solid components, such as red blood cells, white blood cells, platelets, fibrin, etc. These components may clog the pores of the adsorbent, thereby affecting its adsorption effect and perfusion efficiency. 4) Broad-spectrum performance: There are many types of harmful substances in the blood, including organic matter, inorganic matter, drugs, toxins, pathogens, cells, etc. It is a huge challenge to develop an adsorbent that can have efficient adsorption performance for these multiple substances. 5) Adsorption of proteins: Blood contains a large amount of proteins. Some adsorbents may have non-specific adsorption effects on proteins, thereby changing the composition and properties of the blood.

In blood perfusion, the broad-spectrum adsorption of adsorbents mainly includes van der Waals force, hydrogen bond force, electrostatic force, hydrophobic effect and coordination effect, etc. Therefore, surface modification technology can be used to make broad-spectrum adsorbents have multiple forces, which is an effective means to achieve efficient adsorption of multiple substances.

Therefore, based on this, the technical solution of the present invention is proposed.

Summary of the invention

In order to solve the problems existing in the prior art, the present invention provides a method for preparing a blood purification adsorption modified material, the preparation method comprising the following steps:

(1) Soaking the blood purification adsorption material in an oxidant acid solution, reacting and washing, and obtaining a carboxylation purification adsorption material;

(2) First, EDC/NHS-MES buffer is added to the surface of the carboxylated purification adsorption material for reaction, then an amino-containing hydrophobic compound-ethanol/water solution is added for reaction, and finally, NO catalyst solution is added for reaction. After completion, the reaction is washed and dried to obtain a blood purification adsorption modified material.

For ease of understanding of the present invention, the reaction process of the present invention is described:

The method of the present invention first utilizes the oxidation effect of an oxidant to generate carboxyl groups on the surface of a blood purification adsorption material; then, under the action of a carboxyl activator EDC/NHS, a hydrophobic compound is combined with the carboxyl groups on the surface of the adsorption material and a catalyst that can catalyze the decomposition of NO donors to release NO through an amide reaction and a chelation reaction, respectively, and the hydrophobic compound and the catalyst that can catalyze the decomposition of NO donors to release NO are efficiently and firmly fixed on the surface of the adsorption column material by chemical grafting. The increased hydrophobicity of the surface of the blood purification adsorption modified material improves the adsorption capacity of toxins and wastes during the blood purification process; and the catalyst that can catalyze the decomposition of NO donors to release NO can catalyze the endogenous NO donors in the blood to continuously release NO, thereby ensuring the biocompatibility of the blood purification adsorption modified material. The synergistic effect of the hydrophobic compound and the catalyst that can catalyze the decomposition of NO donors to release NO improves the adsorption performance of the blood purification adsorption modified material. The present invention can be applied to the surface modification treatment of blood purification materials and devices.

Among them, the CAS number of NHS is: 6066-82-6, the English name is: N-Hydroxysuccinimide, and the Chinese name is: N-hydroxysuccinimide.

The CAS number of EDC is: 25952-53-8, and its English name is:

1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, Chinese name: 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride.

The CAS number of MES is: 4432-31-9, and its Chinese name is: 2-morpholineethanesulfonic acid.

Preferably, in step (1), the blood purification adsorption material is one of activated carbon resin, cation exchange resin, anion exchange resin, polystyrene, polyoxymethylene, polypropylene, polysulfone, polyethersulfone, nano-activated carbon, nano-graphene, and agarose.

Preferably, in step (1), the oxidant acidic solution is prepared by mixing an oxidant and an acidic aqueous solution;

And/or, the oxidant is one or a combination of two or more of nitric acid, potassium permanganate, and hydrogen peroxide; the concentration of the nitric acid and hydrogen peroxide is 0.5-15%, and the concentration of the potassium permanganate is 0.5-5 mg/mL;

And/or, the pH of the acidic aqueous solution is 3-6.5.

Preferably, in step (1), the reaction temperature is 10-40° C., and the reaction time is 1-48 h.

Preferably, in step (2), the EDC/NHS-MES buffer is prepared by mixing EDC/NHS and MES buffer;

And/or, the amino-containing hydrophobic compound-ethanol/water solution is prepared by mixing an amino-containing hydrophobic compound and an ethanol/water solution;

And/or, the NO catalyst solution is prepared by mixing a catalyst that catalyzes the decomposition of an NO donor to release NO and water.

Preferably, the pH of the MES buffer is 3-6.5;

And/or, in the EDC/NHS-MES buffer, the concentration of EDC is 0.75-7.5 mmol/L, and the concentration of NHS is 0.5-5 mmol/L;

And/or, the amino-containing hydrophobic compound is one or a combination of two or more of hexadecylamine, dodecylamine, N-palmitoylethanolamine, N-oleoylethanolamine, leucine, isoleucine, phenylalanine, polydimethylsiloxane amine, dodecyltrimethylamine, fluorododecylamine, fluorinated polyamine, and poly(p-aminophenylene ether);

and/or, in the amino-containing hydrophobic compound-ethanol/water solution, the concentration of the amino-containing hydrophobic compound is 0.1-10 mg/mL;

and/or, the catalyst for catalyzing the decomposition of the NO donor to release NO is one or a combination of two or more of Cu ²⁺ , Fe ²⁺ , Fe ³⁺ , Mn ²⁺ , Co ²⁺ , Ag ⁺ , ascorbic acid, iron phthalocyanine, cobalt phthalocyanine, ebselen, and selenocystamine;

And/or, in the NO catalyst solution, the concentration of the catalyst for catalyzing the decomposition of the NO donor to release NO is 0.1-10 mg/mL.

Preferably, in step (2), the reaction temperature is 10-40° C., and the reaction time is 1-48 h.

Based on the same technical concept, another embodiment of the present invention is to provide a blood purification adsorption modified material obtained by the above preparation method.

The beneficial effects of the present invention are:

1. The preparation method of the present invention utilizes the oxidation effect of an oxidant in an acidic aqueous solution to generate carboxyl groups in situ on the surface of the blood perfusion adsorbent, thereby converting the inert surface into an active surface. This carboxylation surface process is non-material-dependent, can fully oxidize the surface of the adsorbent and generate carboxyl groups, and will not adversely affect the adsorbent material itself, and has wide applicability and safety.

2. The carboxylated adsorbent undergoes an amide reaction with the hydrophobic compound containing an amine group, fixing the hydrophobic compound molecules on the surface of the material by chemical bonding, significantly improving the hydrophobicity of the adsorbent material without affecting the pore size of the adsorption column.

3. When the NO catalyst chemically fixed on the surface of the adsorbent comes into contact with blood, it will catalyze the decomposition of NO donors in the blood and release NO. As a vascular signaling molecule, NO can effectively inhibit the activation and aggregation of platelets, thereby reducing the formation of thrombi on the surface of the adsorbent and improving blood compatibility; at the same time, NO can inhibit the adhesion and migration of leukocytes, inhibit the release of inflammatory cytokines (such as TNF-α, IL-1, IL-6) and chemokines (such as MCP-1), and has excellent anti-inflammatory effects. Excellent blood compatibility and anti-inflammatory effects can further improve the biocompatibility of the adsorbent, reduce the blockage rate of the adsorbent pores, and improve its adsorption effect and perfusion efficiency.

4. There are multiple forces between the modified adsorbent and the adsorbate, including the comprehensive properties of the adsorbent material itself, hydrophobic effect, coordination effect of NO catalyst, van der Waals force, hydrogen bond and electrostatic force. Two or more of the multiple forces can work together to achieve efficient adsorption of multiple harmful substances in the blood.

5. The method of the present invention is simple to operate, has efficient reaction, does not require expensive equipment, has broad-spectrum practicability, and can fix different hydrophobic compounds and NO catalysts on the surface of the adsorbent to prepare a blood purification adsorption column with excellent adsorption performance.

BRIEF DESCRIPTION OF THE DRAWINGS

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

FIG. 1 is a graph showing the water contact angle of polystyrene before and after modification described in Example 1.

FIG. 2 shows the reaction between the polyoxymethylene adsorption column material described in Example 2 before and after modification and the blood of rabbits with acute kidney injury, and the contents of creatinine (a) and urea nitrogen (b) in the blood.

FIG. 3 is a SEM morphology of the polyethersulfone hemoperfusion adsorption column and microspheres described in Example 3 (a), and the reaction of the polyethersulfone microspheres with platelet-rich plasma before and after modification (b).

FIG4 is the XPS full spectrum (a) and the high-resolution spectrum (b) of the rear surface of the modified polypropylene described in Example 4, and the rate of NO release by the catalytic NO donor (c).

Detailed ways

To make the purpose, technical solution and advantages of the present invention clearer, the technical solution of the present invention will be described in detail below. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other implementation methods obtained by ordinary technicians in this field without creative work belong to the scope of protection of the present invention.

Example 1

This embodiment provides a method for preparing a blood purification adsorption modified material, the preparation method comprising the following steps:

(1) mixing hydrogen peroxide with an acidic aqueous solution of pH=6 to obtain a 5% hydrogen peroxide aqueous solution;

(2) Mixing EDC/NHS with MES buffer at pH = 5 to obtain EDC/NHS-MES buffer; wherein the concentration of EDC is 1 mmol/L and the concentration of NHS is 1.5 mmol/L;

(3) mixing dodecylamine with an ethanol/water solution to obtain a dodecylamine-ethanol/water solution with a concentration of 2 mg/mL; wherein the volume ratio of ethanol to water in the ethanol/water solution is 4:1;

(4) mixing copper chloride with water to obtain a copper chloride solution with a concentration of 1 mg/mL;

(5) fully immersing the blood purification adsorption material polystyrene in a 5% hydrogen peroxide aqueous solution, reacting at 20° C. for 2 hours, and then washing to obtain a carboxylated polystyrene purification adsorption material;

(6) First, EDC/NHS-MES buffer is added to the surface of the carboxylated polystyrene purification adsorption material for reaction, then dodecylamine-ethanol/water solution is added for reaction, and finally copper chloride solution is added for reaction. After completion, the reaction is washed and dried to obtain a blood purification adsorption modified material; the reaction is carried out at 20°C and the total reaction time is 5 hours.

Example 2

This embodiment provides a method for preparing a blood purification adsorption modified material, the preparation method comprising the following steps:

(1) mixing nitric acid with an acidic aqueous solution of pH=3 to obtain a nitric acid aqueous solution with a concentration of 1%;

(2) Mixing EDC/NHS with MES buffer at pH 3 to obtain EDC/NHS-MES buffer; wherein the concentration of EDC is 0.75 mmol/L and the concentration of NHS is 0.5 mmol/L;

(3) mixing isoleucine with an ethanol/water solution to obtain an isoleucine-ethanol/water solution with a concentration of 0.1 mg/mL; wherein the volume ratio of ethanol to water in the ethanol/water solution is 4:1;

(4) mixing ascorbic acid with water to obtain an ascorbic acid solution with a concentration of 0.1 mg/mL;

(5) fully immersing the blood purification adsorption material polyoxymethylene in a 1% nitric acid aqueous solution, reacting at 10° C. for 12 hours, and then washing to obtain a carboxylated polyoxymethylene purification adsorption material;

(6) First, add EDC/NHS-MES buffer to the surface of the carboxylated polyformaldehyde purification adsorption material for reaction, then add isoleucine-ethanol/water solution for reaction, and finally add ascorbic acid solution for reaction. After completion, wash and dry to obtain a blood purification adsorption modified material; the reaction is carried out at 10°C and the total reaction time is 12 hours.

Example 3

This embodiment provides a method for preparing a blood purification adsorption modified material, the preparation method comprising the following steps:

(1) Mixing potassium permanganate with an acidic aqueous solution of pH=6.5 to obtain a potassium permanganate aqueous solution with a concentration of 5 mg/mL;

(2) Mixing EDC/NHS with MES buffer at pH 6.5 to obtain EDC/NHS-MES buffer, wherein the concentration of EDC is 7.5 mmol/L and the concentration of NHS is 5 mmol/L;

(3) mixing dodecyltrimethylamine with an ethanol/water solution to obtain a dodecyltrimethylamine-ethanol/water solution with a concentration of 10 mg/mL; wherein the volume ratio of ethanol to water in the ethanol/water solution is 4:1;

(4) mixing cobalt phthalocyanine with water to obtain a cobalt phthalocyanine solution with a concentration of 10 mg/mL;

(5) fully immersing the blood purification adsorption material polyethersulfone in a potassium permanganate aqueous solution with a concentration of 5 mg/mL, reacting at 40° C. for 1 hour, and then washing to obtain a carboxylated polyethersulfone purification adsorption material;

(6) First, EDC/NHS-MES buffer is added to the surface of the carboxylated polyethersulfone purification adsorption material for reaction, then dodecyltrimethylamine-ethanol/water solution is added for reaction, and finally cobalt phthalocyanine solution is added for reaction. After completion, the reaction is washed and dried to obtain a blood purification adsorption modified material; the reaction is carried out at 40° C. and the total reaction time is 1 hour.

Example 4

This embodiment provides a method for preparing a blood purification adsorption modified material, the preparation method comprising the following steps:

(1) Mixing potassium permanganate with an acidic aqueous solution of pH=6.5 to obtain a potassium permanganate aqueous solution with a concentration of 2 mg/mL;

(2) Mixing EDC/NHS with MES buffer at pH 6.5 to obtain EDC/NHS-MES buffer, wherein the concentration of EDC is 7.5 mmol/L and the concentration of NHS is 5 mmol/L;

(3) mixing the fluorinated polyamine with an ethanol/water solution to obtain a fluorinated polyamine-ethanol/water solution with a concentration of 10 mg/mL; wherein the volume ratio of ethanol to water in the ethanol/water solution is 4:1;

(4) mixing ebselen with water to obtain an ebselen solution with a concentration of 5 mg/mL;

(5) fully immersing the blood purification adsorption material polypropylene in a potassium permanganate aqueous solution with a concentration of 1 mg/mL, reacting at 40° C. for 48 hours, and then washing to obtain a carboxylated polypropylene purification adsorption material;

(6) First, add EDC/NHS-MES buffer to the surface of the carboxylated polypropylene purification adsorption material for reaction, then add fluorinated polyamine-ethanol/water solution for reaction, and finally add ebselen solution for reaction. After completion, wash and dry to obtain a blood purification adsorption modified material; the reaction is carried out at 40° C. and the total reaction time is 48 h.

Verification Example

FIG. 1 is a diagram of water contact angles of polystyrene before and after modification described in Example 1. It can be seen from the diagram that after modification, the water contact of the surface of the adsorption column material is significantly improved.

Figure 2 shows the reaction of the polyformaldehyde adsorption column material before and after modification with the blood of rabbits with acute kidney injury, and the contents of creatinine (a) and urea nitrogen (b) in the blood. It can be seen from the figure that the modified polyformaldehyde material can significantly reduce the contents of creatinine and urea nitrogen in the blood, proving that the modified adsorption column has excellent adsorption performance for toxic substances in the blood.

Figure 3 is a polyethersulfone hemoperfusion adsorption column and microspheres (a) and SEM morphology of polyethersulfone microspheres reacting with platelet-rich plasma before and after modification (b) as described in Example 3. It can be seen from the figure that the modified polyethersulfone still has excellent blood compatibility.

Figure 4 shows the XPS spectrum (a) and the high-resolution spectrum (b) of the modified polypropylene surface, as well as the rate (c) of catalyzing the release of NO by the NO donor. It can be seen from the figure that ebselen, which catalyzes the decomposition of the NO donor and releases NO, has been successfully modified on the surface of the material and can catalyze the release of NO within the physiological concentration range.

The above is only a specific embodiment of the present invention, but the protection scope of the present invention is not limited thereto. Any person skilled in the art who is familiar with the technical field can easily think of changes or substitutions within the technical scope disclosed by the present invention, which should be included in the protection scope of the present invention. Therefore, the protection scope of the present invention should be based on the protection scope of the claims.

Claims (7)

1. A method for preparing a blood purification adsorption modified material, which is characterized by comprising the following steps:

(1) Soaking the blood purification and adsorption material in an oxidant acid solution, and performing reaction and cleaning to obtain a carboxylation purification and adsorption material; wherein:

The blood purification adsorption material is one of activated carbon resin, cation exchange resin, anion exchange resin, polystyrene, polyoxymethylene, polypropylene, polysulfone, polyether sulfone, nano activated carbon, nano graphene and agarose;

(2) EDC/NHS-MES buffer solution is firstly added to the surface of the carboxylation purification adsorption material for reaction, then amine-containing hydrophobic compound-ethanol/water solution is added for reaction, finally NO catalyst solution is added for reaction, and after completion, the material is washed and dried, thus obtaining the blood purification adsorption modified material; wherein:

The NO catalyst solution is prepared by mixing a catalyst for catalyzing NO donor to decompose and release NO with water;

the catalyst for catalyzing the decomposition of the NO donor to release NO is one or a combination of more than two of Cu ²⁺、Fe²⁺、Fe³⁺、Mn²⁺、Co²⁺、Ag⁺, ascorbic acid, iron phthalocyanine, cobalt phthalocyanine, ebselen and selenocysteine;

In the NO catalyst solution, the concentration of a catalyst for catalyzing the decomposition of the NO donor to release NO is 0.1-10 mg/mL.

2. The method for producing a blood purification adsorption-modified material according to claim 1, wherein in the step (1), the oxidizing agent acidic solution is prepared by mixing an oxidizing agent and an acidic aqueous solution;

And/or the oxidant is one or the combination of more than two of nitric acid, potassium permanganate and hydrogen peroxide; the concentration of the nitric acid and the hydrogen peroxide is 0.5-15%, and the concentration of the potassium permanganate is 0.5-5 mg/mL;

and/or the pH value of the acidic aqueous solution is 3-6.5.

3. The method for producing a modified adsorbent for blood purification according to claim 1, wherein in the step (1), the reaction temperature is 10 to 40 ℃ and the reaction time is 1 to 48 hours.

4. The method for preparing a blood purification and adsorption modified material according to claim 1, wherein in the step (2), the EDC/NHS-MES buffer is prepared by mixing EDC/NHS and MES buffer;

and/or the amino-containing hydrophobic compound-ethanol/water solution is prepared by mixing the amino-containing hydrophobic compound and the ethanol/water solution.

5. The method for producing a modified adsorbent for blood purification according to claim 4, wherein the MES buffer has a pH of 3 to 6.5;

and/or EDC/NHS-MES buffer solution, wherein the concentration of EDC is 0.75-7.5 mmol/L, and the concentration of NHS is 0.5-5 mmol/L;

And/or the amino-containing hydrophobic compound is one or more than two of hexadecylamine, dodecylamine, N-palmitoylethanolamine, N-oleoylethanolamide, leucine, isoleucine, phenylalanine, polydimethylsiloxane amine, dodecyltrimethylamine, fluorododecylamine, fluorinated polyamine and poly-p-aminophenether;

and/or, in the amino-containing hydrophobic compound-ethanol/water solution, the concentration of the amino-containing hydrophobic compound is 0.1-10 mg/mL.

6. The method for producing a modified adsorbent for blood purification according to claim 1, wherein in the step (2), the reaction temperature is 10 to 40 ℃ and the reaction time is 1 to 48 hours.

7. The modified blood purification and adsorption material obtained by the method according to any one of claims 1 to 6.