A preparation method of modified blood purification filter membrane and corresponding purifier

Abstract

The present invention discloses a preparation method of a modified blood purification filter membrane and a corresponding purifier, and belongs to the technical field of blood semipermeable membrane separation. The method of the present invention comprises: firstly synthesizing sulfonated polyethersulfone by sulfonation reaction, then synthesizing oxidized cisplatin by oxidation reaction, and then reacting sulfonated polyethersulfone with oxidized cisplatin to prepare sulfonated cisplatin modified blood purification membrane. The sulfonated cisplatin modified blood purification membrane prepared by the chemical grafting method of the present invention has the function of slowly releasing cisplatin, and is particularly suitable for blood purification treatment of tumor patients. The modification process of the present invention is simple and controllable, economical and environmentally friendly, and easy to mass produce.

Description

Technical Field

[0001] The present invention relates to the technical field of blood semipermeable membrane separation, in particular to a method for preparing a modified blood purification filter membrane in contact with blood and a corresponding purifier. Background Art

[0002] Blood purification technology with hemodialysis as the core is increasingly used in clinical practice, especially in the treatment of critical illnesses. Malignant tumors are one of the three major causes of death that pose a serious threat to human health, and their incidence is increasing year by year. The occurrence, development and treatment of tumors are often accompanied by kidney damage and renal failure, requiring blood purification treatment. Cancer patients receiving blood purification treatment constitute a special group, who usually have the characteristics of severe underlying diseases, complex conditions, low immunity and poor prognosis. Although blood purification technology has made significant progress, its treatment is still "invasive". When cancer patients receive blood purification, the incidence of complications such as thrombosis, infection and allergy is significantly higher than that of other patients. Therefore, it is very necessary to minimize the "damaging" treatment methods (such as chemotherapy, radiotherapy and surgery) for cancer patients when the condition permits.

[0003] Cisplatin is one of the most commonly used anti-tumor chemotherapy drugs, widely used in the treatment of various malignancies such as lung cancer, ovarian cancer, prostate cancer, testicular cancer, nasopharyngeal cancer and esophageal cancer. The structure of cisplatin is centered on a divalent platinum ion, with two amino groups as ligands and two chloride ions as leaving groups. In the blood or extracellular fluid, the physiological concentration of chloride ions is about 100mmol / L, and the activity of cisplatin is low at this time; but when cisplatin enters the cell, the intracellular chloride concentration drops to several mmol / L. (The activity of cisplatin is usually significantly enhanced when the chloride concentration is lower than 20mmol / L in the cell. In experiments, the intracellular environment is often simulated, and it is believed that cisplatin has the strongest effect when the chloride concentration is in the range of 4-10mmol / L, and can efficiently bind to target molecules (such as DNA). The amino acid side chains, DNA and RNA in the cell contain a large number of nucleophilic groups (such as oxygen, nitrogen, thiol, etc.) that can undergo replacement reactions with the chloride ions in cisplatin to form new complexes. This process will disrupt DNA replication and transcription, thereby exerting its anti-tumor effect.

[0004] Completing blood purification treatment requires a blood purification filter composed of a hollow fiber filter membrane. At present, most blood purification membranes are artificially synthesized from polymer materials. Today, the modification of blood purification filter membrane materials is mainly focused on improving their anticoagulant, anti-inflammatory properties and biocompatibility. For example, heparin or heparin-like molecules are coated on dialysis membrane materials to give them anticoagulant activity; Avacopan is grafted on dialysis membrane materials to enhance their anti-inflammatory ability; and albumin is grafted on dialysis membrane materials to improve their biocompatibility. However, no research reports on blood purification filter membranes with anti-tumor effects have been found. Summary of the invention

[0005] The technical problem solved by the present invention is that: the risk of tumor patients receiving blood purification treatment is high, and there is no blood purification filter membrane specifically for tumor patients; the local modification method of the blood purification filter membrane is relatively complicated, the reaction conditions are harsh, and it is not conducive to industrial production.

[0006] Specifically, on the one hand, the present invention provides a method for preparing a modified blood purification filter membrane, wherein the modified blood purification filter membrane is a sulfonated cisplatin-modified polyethersulfone blood purification filter membrane, and the method comprises:

[0007] (1) dissolving the dried polyethersulfone in concentrated sulfuric acid, stirring well, and adding chlorosulfonic acid to react, wherein the mass ratio of polyethersulfone to chlorosulfonic acid is 10:1 to 2:1. After the reaction is completed, repeatedly washing the reaction product of this step with double distilled water, and vacuum drying the reaction product of this step to constant weight to obtain sulfonated polyethersulfone;

[0008] (2) dissolving cisplatin in distilled water, adding 30% hydrogen peroxide to react, and after the reaction is completed, repeatedly washing the reaction product of this step with double distilled water, and vacuum drying to constant weight to obtain oxidized cisplatin, wherein the mass ratio of polyethersulfone in step (1) to cisplatin in this step is 4:3 to 4:1;

[0009] (3) dissolving the reaction products obtained in step (1) and step (2) in dichloromethane for reaction, wherein the mass ratio of the two reaction products is 2:1 to 1:2, repeatedly washing the reaction product of this step with double distilled water, and vacuum drying to constant weight to obtain sulfonated cisplatin modified polyether sulfone;

[0010] (4) Dissolving sulfonated cisplatin-modified polyethersulfone and polyethersulfone in N-methylpyrrolidone solvent to prepare a 10%-20% solution, wherein the mass ratio of sulfonated cisplatin-modified polyethersulfone to polyethersulfone is 1:1 to 1:6, and using an immersion precipitation phase conversion method to prepare a sulfonated cisplatin-modified polyethersulfone blood purification filter membrane.

[0011] In a preferred implementation, the mass of polyethersulfone in step (1) is 10 g, and the amount of chlorosulfonic acid used is 1 mL.

[0012] In another preferred implementation, the mass ratio of polyethersulfone in step (1) to cisplatin in step (2) is 10:3.

[0013] In another preferred implementation, in step (1), the reaction temperature is 0°C-30°C, and the reaction time is 0.5-3h;

[0014] In step (2), the reaction temperature is 10°C-50°C, and the reaction time is 1-2h;

[0015] In step (3), the reaction temperature is 20°C-30°C, and the reaction time is 3-30h.

[0016] On the other hand, the present invention provides a purifier for purifying blood, wherein the purifier adopts the modified blood purification filter membrane prepared by the method.

[0017] On the other hand, the present invention provides an application of the modified blood purification filter membrane, wherein the sulfonated cisplatin modified blood purification filter membrane is installed in a hemodialysis device.

[0018] On the other hand, the present invention provides an anti-tumor biomaterial of a modified blood purification filter membrane prepared by the method, wherein the biomaterial comprises a sulfonic acid molecule layer grafted on the biomaterial and cisplatin grafted on the sulfonic acid group.

[0019] Preferably, the biomaterial is a water-insoluble polymer material, including at least one of polysulfone, polyethersulfone, polyacrylonitrile, polyolefin and polyvinyl alcohol.

[0020] Compared with the prior art, the present invention has the following beneficial effects:

[0021] The preparation method of the present invention first prepares sulfonated polyethersulfone, oxidizes cisplatin, and then reacts the sulfonated polyethersulfone with oxidized cisplatin, thereby grafting cisplatin on the polyethersulfone membrane. During blood purification treatment, cisplatin dissociates from the polyethersulfone membrane, and simultaneously plays a role in blood purification and anti-tumor. The present invention proposes for the first time a method for preparing a blood purification filter membrane with anti-tumor effect, which is particularly suitable for blood purification treatment of tumor patients.

[0022] The present invention has mild reaction conditions, low cost of required polymer materials, reagents and drugs, economic and environmental protection; the whole modification process is simple and easy, suitable for industrial production and easy to promote. The modified blood purification filter membrane also has anti-tumor effect and has great application prospects.

[0023] The blood purification filter membrane prepared by the method provided by the present invention does not release in a non-reducing environment, but can effectively release in a special reducing microenvironment of tumor cells, and has a strong prospect for targeted treatment.

[0024] In the purification filter membrane of the present invention, the grafted drug molecules still have good anti-tumor effects, and the bulk properties of the polymer material are not significantly affected. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 Schematic diagram showing the anti-tumor mechanism of sulfonated cisplatin-modified polyethersulfone blood purification filter membrane.

[0026] Figure 2 The following is a reaction formula for preparing sulfonated polyethersulfone.

[0027] Figure 3 The preparation of sulfonated cisplatin-modified polyethersulfone blood purification filter membrane and the cisplatin release reaction formula are shown.

[0028] Figure 4 The graph shows the drug release curves of the sulfonated cisplatin modified polyethersulfone blood purification filter membrane under different environments. DETAILED DESCRIPTION

[0029] The present invention will be further described below in conjunction with the embodiments.

[0030] The following embodiments of the present invention are described by taking a blood purification filter membrane as an example.

[0031] In the following examples, blood purification filter membranes with anti-tumor properties were prepared using exemplary ratios, but the technical solution of the present invention is applicable to other similar solutions, and these solutions are included in the scope of the present invention.

[0032] Example 1

[0033] ① Weigh 10g of dried polyethersulfone (PES) and dissolve it in 30mL of concentrated sulfuric acid, then slowly drop 1mL of chlorosulfonic acid (ClHSO3). Adjust the temperature of the reaction system to 0℃ and react for 1h. After the reaction is completed, cool it to room temperature and slowly pour the solution into double distilled water to obtain a brown solid. Wash it repeatedly with double distilled water to remove the residual solvent, dry it until it is neutral, and obtain sulfonated polyethersulfone.

[0034] ② Take 3g of cisplatin and dissolve it in 200mL of double distilled water. Add 150mL of 30% hydrogen peroxide (10 times excess) while stirring. Adjust the temperature of the reaction system to 20℃ and react for 1h in the dark. When the solution turns light yellow and dissolves, filter it while hot, then place the filtrate in a 4℃ refrigerator for recrystallization for about 4h, filter it, and wash it with cold water, cold ether, and cold ethanol for about three times. Finally, pale yellow crystals of oxidized cisplatin are obtained. The sample is vacuum dried in the dark.

[0035] ③ Dissolve 1 g of sulfonated polyethersulfone and 0.5 g of oxidized cisplatin in 20 mL of dichloromethane, adjust the temperature of the reaction system to 20°C, and react for 6 hours in the dark. After the reaction is completed, cool to room temperature, wash the reaction product repeatedly with double distilled water, and dry it in vacuum at 60°C to obtain sulfonated cisplatin-modified polyethersulfone.

[0036] ④ Take 0.2g of sulfonated cisplatin modified polyethersulfone and 1g of polyethersulfone and dissolve them in 5.5mL of N-methylpyrrolidone solvent to make an 18% solution. Use the immersion precipitation phase conversion method to drop the solution onto a carrier such as a glass plate, scrape it into a film, and place it in a coagulation bath (water) to make a sulfonated cisplatin modified polyethersulfone blood purification filter membrane.

[0037] Example 2

[0038] ① Weigh 10g of dried polyethersulfone (PES) and dissolve it in 30mL of concentrated sulfuric acid, then slowly drop 1.5mL of chlorosulfonic acid (ClHSO3). Adjust the temperature of the reaction system to 0℃ and react for 2h. After the reaction is completed, cool it to room temperature and slowly pour the solution into double distilled water to obtain a brown solid. Wash it with double distilled water several times to remove the residual solvent, dry it until it is neutral, and obtain sulfonated polyethersulfone.

[0039] ② Take 3g of cisplatin and dissolve it in 200mL of double distilled water. Add 150mL of 30% hydrogen peroxide (10 times excess) while stirring. Adjust the temperature of the reaction system to 30℃ and react for 1h in the dark. When the solution turns light yellow and dissolves, filter it while hot, then place the filtrate in a 4℃ refrigerator for recrystallization for about 4h, filter it, and wash it with cold water, cold ether, and cold ethanol for about three times. Finally, pale yellow crystals of oxidized cisplatin are obtained. The sample is vacuum dried in the dark.

[0040] ③ Take 1g of sulfonated polyethersulfone and 1g of oxidized cisplatin and dissolve them in 20mL of dichloromethane, adjust the temperature of the reaction system to 25℃, and react for 12h in the dark. After the reaction is completed, cool to room temperature, wash the reaction product repeatedly with double distilled water, and dry it in vacuum at 60℃ to obtain sulfonated cisplatin modified polyethersulfone.

[0041] ④ Take 0.5g of sulfonated cisplatin modified polyethersulfone and 1g of polyethersulfone and dissolve them in 6.8mL of N-methylpyrrolidone solvent to prepare an 18% solution, and use the immersion precipitation phase conversion method to prepare the sulfonated cisplatin modified polyethersulfone blood purification filter membrane.

[0042] Example 3

[0043] ① Weigh 10g of dried polyethersulfone (PES) and dissolve it in 30mL of concentrated sulfuric acid, then slowly drop 1mL of chlorosulfonic acid (ClHSO3). Adjust the temperature of the reaction system to 30℃ and react for 3h. After the reaction is completed, cool it to room temperature and slowly pour the solution into double distilled water to obtain a brown solid. Wash it with double distilled water several times to remove the residual solvent, dry it until it is neutral, and obtain sulfonated polyethersulfone.

[0044] ② Take 3g of cisplatin and dissolve it in 200mL of double distilled water. Add 150mL of 30% hydrogen peroxide (10 times excess) while stirring. Adjust the temperature of the reaction system to 50℃ and react for 2h in the dark. When the solution turns light yellow and dissolves, filter it while hot, then place the filtrate in a 4℃ refrigerator for recrystallization for about 4h, filter it, and wash it with cold water, cold ether, and cold ethanol for about three times. Finally, pale yellow crystals of oxidized cisplatin are obtained. The sample is vacuum dried in the dark.

[0045] ③ Dissolve 1g of sulfonated polyethersulfone and 2g of oxidized cisplatin in 20mL of dichloromethane, adjust the temperature of the reaction system to 25°C, and react for 24h in the dark. After the reaction is completed, cool to room temperature, wash the reaction product repeatedly with double distilled water, and dry it in vacuum at 60°C to obtain sulfonated cisplatin modified polyethersulfone.

[0046] ④ Take 0.8g of sulfonated cisplatin modified polyethersulfone and 1g of polyethersulfone and dissolve them in 8.2mL of N-methylpyrrolidone solvent to prepare an 18% solution, and use the immersion precipitation phase conversion method to prepare the sulfonated cisplatin modified polyethersulfone blood purification filter membrane.

[0047] Comparative Example 1:

[0048] Weigh 10g of dried polyethersulfone (PES) and dissolve it in 30mL of concentrated sulfuric acid, then slowly drop 2mL of chlorosulfonic acid (ClHSO3). Adjust the temperature of the reaction system to 60℃ and react for 1h. After the reaction is completed, cool it to room temperature and slowly pour the solution into double distilled water to obtain a black solid. Repeat the double distilled water washing several times to remove the residual solvent to obtain a black solid, indicating that a carbonization reaction has occurred and the reaction has failed. This indicates that the temperature of the sulfonation process cannot be too high.

[0049] Comparative Example 2:

[0050] Weigh 10g of dried polyethersulfone (PES) and dissolve it in 50mL of concentrated sulfuric acid, then slowly drop 1mL of chlorosulfonic acid (ClHSO3). Adjust the temperature of the reaction system to 0℃ and react for 1h. After the reaction is completed, cool it to room temperature and slowly pour the solution into double distilled water. Wash it repeatedly with double distilled water several times to remove the residual solvent, and obtain a gray solid substance, which is a mixture of sulfonated polyethersulfone and polyethersulfone, indicating that excessive concentrated sulfuric acid inhibits the reaction and reduces the yield.

[0051] The applicant tested the products prepared in the examples of the present invention.

[0052] The product of Example 3 was used to test the release behavior of Pt in the PES-Pt polymer drug system prepared by the present invention in a reducing environment of a PBS solution containing 1 mM ascorbic acid and a non-reducing environment of a PBS solution.

[0053] like Figure 4 As shown, over time, the inventors observed that PES-Pt nanomedicines are almost stable in PBS solutions. After 40 hours of release, only about 5.4% of platinum drugs were released, and they remained stable after 16 hours; in the release results in PBS solutions containing 1mM ascorbic acid, it can be seen that PES-Pt drugs have reduction-sensitive release properties. PES-Pt is rapidly and continuously released in 1mM ascorbic acid PBS solution. After 40 hours, the release amount of platinum drugs released from PES-Pt also reaches 24.6%, indicating that PES-Pt polymer drugs can effectively release platinum drugs in the simulated tumor cell reduction microenvironment. This result proves that PES-Pt polymer drugs have ascorbic acid reduction responsiveness and can continuously release chemotherapy cisplatin drugs in the special reduction environment of tumor cells.

[0054] The present invention adopts a chemical grafting method to graft cisplatin onto a polyethersulfone blood purification filter membrane. When the sulfonated cisplatin modified polyethersulfone membrane contacts blood and enters cells, the Pt(IV) complex will undergo two electron reduction reactions under the action of a cell reducing agent in the special reduction environment of tumor cells, thereby regenerating the original square plane Pt(II) cisplatin drug and releasing two polyethersulfone ligands. Cisplatin is dissociated from the membrane material and slowly released, allowing tumor patients to receive anti-tumor treatment while undergoing blood purification. The sulfonated cisplatin modified blood purification filter membrane has both blood purification function and anti-tumor effect, thereby reducing the number of "damaging" treatments and medical burdens for tumor patients, achieving multiple goals at one stroke.

[0055] The above is only a preferred specific implementation manner of the present invention, and the protection scope of the present invention is not limited thereto. Any simple change or equivalent replacement of the technical solution that can be obviously obtained by any technician familiar with the technical field within the technical scope disclosed in the present invention falls within the protection scope of the present invention.

Claims

1. A method for preparing a modified blood purification filter membrane, characterized in that: The modified blood purification filter membrane is a sulfonated cisplatin modified polyethersulfone blood purification filter membrane, and the method comprises: (1) dissolving the dried polyethersulfone in concentrated sulfuric acid, stirring well, and adding chlorosulfonic acid to react, wherein the mass ratio of polyethersulfone to chlorosulfonic acid is 10:1 to 2:

1. After the reaction is completed, repeatedly washing the reaction product of this step with double distilled water, and vacuum drying the reaction product of this step to constant weight to obtain sulfonated polyethersulfone; (2) dissolving cisplatin in distilled water, adding 30% hydrogen peroxide to react, and after the reaction is completed, repeatedly washing the reaction product of this step with double distilled water, and vacuum drying to constant weight to obtain oxidized cisplatin, wherein the mass ratio of polyethersulfone in step (1) to cisplatin in this step is 4:3 to 4:1; (3) dissolving the reaction products obtained in step (1) and step (2) in dichloromethane for reaction, wherein the mass ratio of the two reaction products is 2:1 to 1:2, repeatedly washing the reaction product of this step with double distilled water, and vacuum drying to constant weight to obtain sulfonated cisplatin modified polyether sulfone; (4) Dissolving sulfonated cisplatin-modified polyethersulfone and polyethersulfone in N-methylpyrrolidone solvent to prepare a 10%-20% solution, wherein the mass ratio of sulfonated cisplatin-modified polyethersulfone to polyethersulfone is 1:1 to 1:6, and using an immersion precipitation phase conversion method to prepare a sulfonated cisplatin-modified polyethersulfone blood purification filter membrane.

2. The method for preparing the modified blood purification filter membrane according to claim 1, characterized in that: The mass of polyethersulfone in step (1) is 10 g, and the amount of chlorosulfonic acid is 1 mL.

3. The method for preparing the modified blood purification filter membrane according to claim 1, characterized in that: The mass ratio of polyethersulfone in step (1) to cisplatin in step (2) is 10:

3.

4. The method for preparing the modified blood purification filter membrane according to claim 1, characterized in that: In step (1), the reaction temperature is 0°C-30°C, and the reaction time is 0.5-3h; In step (2), the reaction temperature is 10°C-50°C, and the reaction time is 1-2h; In step (3), the reaction temperature is 20°C-30°C, and the reaction time is 3-30h.

5. A purifier, characterized in that: The purifier is used for purifying blood, and the purifier adopts the modified blood purification filter membrane prepared by the method of claim 1.

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