CN116173175A

CN116173175A - Preparation method of blood perfusion filler for treating immunodeficiency-induced tumors

Info

Publication number: CN116173175A
Application number: CN202310163791.9A
Authority: CN
Inventors: 李昱锟; 周友浪; 陈情忠
Original assignee: Jiangsu Qirui Biotechnology Co ltd
Current assignee: Jiangsu Qirui Biotechnology Co ltd
Priority date: 2023-02-24
Filing date: 2023-02-24
Publication date: 2023-05-30

Abstract

The invention relates to the technical field of biomedical science, in particular to a preparation method of a blood perfusion filler for treating tumors caused by immunodeficiency, which is formed by mixing a black bean peptide solution and an inner core according to the following steps: respectively preparing black bean peptide solution and an inner core; a. preparing black bean peptide solution; adding 6-10 parts of black bean protein hydrolysate and 3-5 parts of enzyme into 85-95 parts of water according to the mass ratio, and carrying out enzymolysis on the black bean protein hydrolysate at the temperature of 50-60 ℃ for 60-80 min to obtain hydrolysate containing black bean peptide and amino acid; the blood perfusion filler prepared from the immunocompetent peptide can well regulate immune organs of a human body by filling the filler into a blood perfusion device, so that the blood of tumors caused by immunodeficiency can be purified, and the toxic and side effects of the medicine can be reduced.

Description

Preparation method of blood perfusion filler for treating immunodeficiency-induced tumors

Technical Field

The invention relates to the technical field of biomedical technology, in particular to a preparation method of a blood perfusion filler for treating immunodeficiency-induced tumors.

Background

At present, the probability of diseases caused by low immunity rises year by year, but the probability of diseases caused by low immunity is an invisible problem. And when the medicine is used for carrying out immune conditioning, other diseases can be caused by taking the medicine for a long time. In recent years, tumor diseases have become one of the main causes of life threatening for human beings, but most of the current tumor therapeutic drugs have immunosuppressive effects, and if used for a long time, the drugs damage the immune system and cause other toxic and side effects. Therefore, it is important to develop safer and more efficient immunotherapeutic approaches. To date, the main methods of treating tumors still have certain limitations. For example, chemotherapeutic agents, which lack judgment on healthy cells in the treatment of tumors, both kill malignant cells and inevitably damage healthy cells, thereby producing toxic side effects. The tumor is generated in the host body mainly because the anti-tumor mechanism loses activity, and the immune response of the body can be improved to enhance the activity of the anti-tumor mechanism. Therefore, some immunoactive peptides with immunoregulatory effects can play a role as immunoregulatory agents in anti-tumor mechanisms. The research shows that the anti-tumor mechanism can enhance the immunoregulation effect through the bioactive peptide in different levels and stages in different forms, for example, the bioactive peptide extracted from plant protein or milk casein can enhance the immunoregulation capability of the organism, and has the characteristics of enhancing lymphocyte transformation and NK cell activity, thereby enhancing the immunity of the organism to pathogens in the external environment and reducing the incidence of tumors.

The current scientific research shows that the immune active peptide has important significance for enhancing the disease prevention capability and improving the immunity resistance capability of human bodies to external pathogens, has the advantages of no toxicity, high safety and the like, and has the active functions of antioxidation, anti-tumor, immunoregulation and the like along with the research discovery. Because the molecular mass of the immune active peptide is small, the rejection reaction of the organism is not caused, and the development of the active peptide with novel functions is gradually approaching to the direction of the immune peptide. Since a means for delivering a protein peptide into a human body by blood perfusion has not been found so far, the use of a protein peptide is of great importance for effective alleviation of diseases caused by immunodeficiency and supplementary therapy by attaching the protein peptide to a resin.

Disclosure of Invention

In order to realize the blood purification of immunodeficiency-induced tumors and reduce the toxic and side effects of medicines, the application provides a preparation method of a blood perfusion filler for treating immunodeficiency-induced tumors.

The preparation method of the blood perfusion filler for treating immunodeficiency-induced tumors adopts the following technical scheme:

the preparation method of the hemoperfusion filling material for treating immunodeficiency-induced tumors comprises the following steps of mixing a black bean peptide solution and an inner core:

step one, respectively preparing a black bean peptide solution and an inner core;

a. preparation of Black Soy peptide solution

Firstly, adding 6-10 parts of black bean protein hydrolysate and 3-5 parts of enzyme into 85-95 parts of water according to the mass ratio, and carrying out enzymolysis on the black bean protein hydrolysate at the temperature of 50-60 ℃ for 60-80 min to obtain hydrolysate containing black bean peptide and amino acid;

secondly, freeze-drying the hydrolysate to obtain freeze-dried powder;

finally, in mass (mg/kg): taking freeze-dried powder with the volume (ml) of 1-4:9, and adding the freeze-dried powder into PBS buffer solution to obtain black soybean peptide solution;

b. preparation of inner core

Taking polystyrene resin as a carrier of an inner core, and performing biological gel: uniformly coating the biological glue on the surface of the polystyrene resin according to the mass ratio of polystyrene resin=1:4-5 to obtain an inner core;

step two, mixing a black bean peptide solution and an inner core;

pouring the inner core into the prepared black bean peptide solution, and filtering out after the inner core and the black bean peptide solution are fully and uniformly mixed to obtain a semi-finished product;

and drying the semi-finished product to obtain a finished product, namely the blood perfusion filler.

Through adopting the steps, the immune active peptide-black bean peptide in the black bean protein is decomposed and extracted, the immune active peptide is adopted to prepare the blood perfusion filler, the blood perfusion filler is contacted with blood, the immune regulation capability of the immune active peptide is utilized, the incidence rate of tumors can be reduced, the immune active peptide is attached to the blood perfusion filler and is sent into a body, and the immune regulation capability of the body is enhanced to different degrees, so that the immune organ of the body can be well regulated after perfusion, and the toxic and side effects caused by adopting drug treatment are avoided.

Preferably, in the first step, the bio-glue is uniformly coated on the polystyrene resin by adopting an atomization spraying mode.

By adopting the technical scheme, the surface of the polystyrene resin can be ensured to fully absorb the biological glue.

Preferably, in the first step, when the black bean peptide is dissolved in the PBS buffer, the PBS buffer floods the polystyrene resin.

Preferably, in the second step, the polystyrene resin and the black bean peptide solution are fully and uniformly mixed by adopting a shaking table, and the shaking time is more than 2 hours.

Preferably, in the second step, the polystyrene resin is poured into a dehydrator for drying.

Preferably, in the first step, the black bean peptide solution is adopted to prepare freeze-dried powder at the temperature of minus 50 ℃ to minus 80 ℃ by a freeze dryer.

Preferably, the biological glue adopts alpha-cyano octyl acrylate.

In summary, the present application has the following beneficial effects:

the blood perfusion filler prepared from the immunocompetent peptide can well regulate immune organs of a human body by filling the filler into a blood perfusion device, so that the blood of tumors caused by immunodeficiency can be purified, and the toxic and side effects of the medicine can be reduced.

Drawings

FIG. 1 shows the morphological changes of spleen tissue of immunodeficiency New Zealand rabbits (200X);

FIG. 2 shows the change in thymus histomorphology of an immunodeficient New Zealand rabbit (200X);

FIG. 3 is a graph showing the results of comparison of the effect of black bean peptide on proliferation of immunodeficient New Zealand rabbit lymphocytes;

FIG. 4 is a graph of the results of a comparison of black bean peptide versus immunodeficiency New Zealand rabbit NK cell activity;

FIG. 5 is a graph showing the results of phagocytic function of black bean peptide on macrophages in the abdominal cavity of an immunodeficient New Zealand rabbit.

Detailed Description

The following description of the embodiments of the present invention will be made clearly and completely, and it is apparent that the described embodiments are only some embodiments of the present invention, but not all embodiments.

The preparation method comprises the steps of preparing a blood perfusion filler, wherein the blood perfusion filler is formed by mixing a black bean peptide solution and an inner core according to the following steps:

example 1:

in the preparation of the black bean peptide solution, 5 parts of black bean protein hydrolysate and 3 parts of enzyme are added into 85 parts of water according to the mass ratio, and the black bean protein hydrolysate is subjected to enzymolysis at 60 ℃ for 70min, so that the hydrolysate containing the black bean peptide and the amino acid is obtained. The hydroxyl radical removal rate was 31.92% at this time. The content of black bean protein peptide is 84.36%. The amino acid species lacks threonine.

Example 2:

in the preparation of the black bean peptide solution, 7 parts of black bean protein hydrolysate and 4 parts of enzyme are added into 89 parts of water according to the mass ratio, and the black bean protein hydrolysate is subjected to enzymolysis at 60 ℃ for 70min, so that the hydrolysate containing the black bean peptide and the amino acid is obtained. The hydroxyl radical removal rate at this time was 32.85%. The content of black bean protein peptide under the optimal hydrolysis condition is 85.02 percent. The amino acids are complete in variety and rich in content, wherein the ratio of essential amino acids to branched chain amino acids in the total content is high, and threonine and glycine in the black bean peptide have the effect of regulating and controlling the immune function.

Example 3:

in the preparation of the black bean peptide solution, 10 parts of black bean protein hydrolysate and 5 parts of enzyme are added into 95 parts of water according to the mass ratio, and the black bean protein hydrolysate is subjected to enzymolysis at 60 ℃ for 70min, so that the hydrolysate containing the black bean peptide and the amino acid is obtained. At this time, the OH removal rate was 30.95%. The content of black bean protein peptide is 84.75%. The amino acid species lacks glycine.

Comparative example 1:

in the preparation of the black bean peptide solution, 3 parts of black bean protein hydrolysate and 2 parts of enzyme are added into 80 parts of water according to the mass ratio, and the black bean protein hydrolysate is subjected to enzymolysis at 60 ℃ for 70min, so that the hydrolysate containing the black bean peptide and the amino acid is obtained. The hydroxyl radical removal rate at this time was 27.65%. The content of black bean protein peptide is 74.76%. The amino acid species lacks glutamic acid.

Comparative example 2:

in the preparation of the black bean peptide solution, 10 parts of black bean protein hydrolysate and 6 parts of enzyme are added into 98 parts of water according to the mass ratio, and the black bean protein hydrolysate is subjected to enzymolysis at 60 ℃ for 70min, so that the hydrolysate containing the black bean peptide and the amino acid is obtained. The hydroxyl radical removal rate at this time was 26.03%. The content of black bean protein peptide is 73.36%. The amino acid species lacks arginine.

In summary, examples 1 to 3 and comparative examples 1 to 2 show that the optimal enzymolysis conditions for preparing the black bean protein hydrolysate are obtained by adding 7 parts of the black bean protein hydrolysate and 4 parts of the enzyme into 89 parts of water in a mass ratio and carrying out enzymolysis on the black bean protein hydrolysate at a temperature of 60 ℃ for 70 minutes. Through molecular weight distribution analysis, the black bean peptide has a molecular weight of 2079-658 Da accounting for 55.17% of the main component. The peptide with molecular weight less than 658Da accounts for 0.91%. From this, it is speculated that black bean peptide has potential immunoregulatory effect.

Example 4:

the blood perfusion filler is prepared in the embodiment, and is formed by mixing a black bean peptide solution and an inner core according to the following steps:

in the preparation of the black bean peptide solution, 7 parts of black bean protein hydrolysate and 4 parts of enzyme are added into 89 parts of water according to the mass ratio, and the black bean protein hydrolysate is subjected to enzymolysis at 60 ℃ for 70min to obtain hydrolysate containing black bean peptide and amino acid; secondly, freeze-drying the hydrolysate to obtain freeze-dried powder; finally, according to the mass: and adding the freeze-dried powder into PBS buffer solution with the volume of 1:9 to obtain a black bean peptide solution, wherein the ratio is obtained through gastric lavage measurement, and the ratio of the black bean peptide solution is lower than the ratio, so that the protein peptide is not effective, and immunosuppression can occur above the ratio.

When the inner core is prepared, taking polystyrene resin as a carrier of the inner core, and uniformly coating 8 parts of biological glue on the surface of 40 parts of polystyrene resin according to the mass ratio to obtain the inner core; the bio-glue in this example is n-octyl α -cyanoacrylate.

Mixing the black bean peptide solution and the inner core; in the embodiment, 200mg/kg of freeze-dried powder is taken according to 1kg of animal weight and added into 1800ml of buffer solution to prepare a low-concentration black bean peptide solution, the inner core is poured into the prepared black bean peptide solution, and the black bean peptide solution is filtered out after the inner core and the black bean peptide solution are fully and uniformly mixed to obtain a semi-finished product; and drying the semi-finished product to obtain a finished product, namely the blood perfusion filler, and completing the establishment of the drug model in the embodiment.

Example 5:

in the preparation of the black bean peptide solution, 8 parts of black bean protein hydrolysate and 3 parts of enzyme are added into 80 parts of water according to the mass ratio, and the black bean protein hydrolysate is subjected to enzymolysis at 60 ℃ for 70min to obtain hydrolysate containing black bean peptide and amino acid; secondly, freeze-drying the hydrolysate to obtain freeze-dried powder; the weight is as follows: and adding the freeze-dried powder into PBS buffer solution to obtain the black soybean peptide solution with the volume of 2:9.

Mixing the black bean peptide solution and the inner core; in the embodiment, according to the weight of 1kg of animals, 400mg/kg of freeze-dried powder is added into 1800ml of buffer solution to prepare a medium-concentration black bean peptide solution, the inner core is poured into the prepared black bean peptide solution, and the black bean peptide solution is filtered out after the inner core and the black bean peptide solution are fully and uniformly mixed to obtain a semi-finished product; and drying the semi-finished product to obtain a finished product, namely the blood perfusion filler, and completing the establishment of the drug model in the embodiment.

Example 6:

the blood perfusion filler is prepared in the embodiment, and is formed by mixing black bean peptide solution and an inner core according to the following steps,

in the preparation of the black bean peptide solution, 8 parts of black bean protein hydrolysate and 3 parts of enzyme are added into 80 parts of water according to the mass ratio, and the black bean protein hydrolysate is subjected to enzymolysis at 60 ℃ for 70min to obtain hydrolysate containing black bean peptide and amino acid; secondly, freeze-drying the hydrolysate to obtain freeze-dried powder; finally, according to the mass: and adding the freeze-dried powder into PBS buffer solution to obtain the black soybean peptide solution with the concentration of 50 percent.

Mixing the black bean peptide solution and the inner core; in the embodiment, according to the weight of 1kg of animal, 800mg/kg of freeze-dried powder is added into 1800ml of buffer solution to prepare high-concentration black bean peptide solution, the inner core is poured into the prepared black bean peptide solution, and the black bean peptide solution is filtered out after the inner core and the black bean peptide solution are fully and uniformly mixed to obtain a semi-finished product; and drying the semi-finished product to obtain a finished product, namely the blood perfusion filler, and completing the establishment of the drug model in the embodiment.

In examples 4-6, it can be seen that the black bean peptide was formulated with the inner core according to low, medium and high dosages, respectively, as seen in C, D, E of FIGS. 1 and 2, and the effect of the black bean peptide on immune function was observed by modeling animal immunodeficiency in examples 4-6 described above.

Most immunodeficient animal model studies select rodents as modeled animals, such as rats, mice. The big and small mice are experimental animals used conventionally, but are not suitable for experimental study of blood perfusion due to small weight and small total blood volume of the whole body. Adult New Zealand rabbits are also experimental animals which are used conventionally, no research report on an alcoholic liver injury model of the New Zealand rabbits exists in the past, and the weight of the adopted New Zealand rabbits is generally 3-3.5kg. The composition has high weight, and can be used for blood perfusion treatment. The present example thus establishes an animal model of alcoholic liver injury for blood perfusion treatment in New Zealand rabbits.

New Zealand rabbits were sampled and preserved before the perfusion experiment was started, and pre-experimental preparation was performed before the start of the experiment, with 10mL of cyclophosphamide reagent at a dose of 80mg/kg being injected into each New Zealand rabbit, and the normal control group was injected with an equal amount of sterile physiological saline at a concentration of 0.9%. As shown in fig. 1 and 2, new zealand rabbits were adaptively raised and fed with standard feed in an experimental environment of (24±2) °c and a relative humidity of (45±3)%, and after 7 days, they were divided into a normal control group, an immunodeficiency group and a black bean peptide experimental group according to body weight, and the black bean peptide experimental groups were respectively black bean peptide low, medium and high dose groups, and their corresponding black bean peptide masses were 200, 400 and 800mg/kg, respectively. As cyclophosphamide is used as a clinically common immunosuppression drug, the cyclophosphamide can cause the reduction of the immune function of a host and is commonly used as an experimental modeling drug. Thus, this study used cyclophosphamide to construct an animal model of immunodeficiency.

In the above embodiment, the bio-glue is uniformly coated on the polystyrene resin by adopting an atomization spraying mode, so that the surface of the polystyrene resin can be ensured to fully absorb the bio-glue. When the black bean peptide was dissolved in the PBS buffer, the PBS buffer submerged the polystyrene resin. And (3) fully and uniformly mixing the polystyrene resin and the black bean peptide solution by adopting a shaking table, wherein the shaking time is more than 2 hours. Freezing the black bean peptide solution at a low temperature of between 50 ℃ below zero and 80 ℃ below zero, and crushing the frozen zymolyte into freeze-dried powder. And pouring the polystyrene resin into a dehydrator for spin-drying to obtain the filler.

The black bean peptide has good immunoregulatory function, and the prepared filler can well regulate immune organs of organisms after blood perfusion in vivo. The clearance of hydroxyl groups of the enzymatic hydrolysis black bean protein hydrolysate is 32.85%, so that the content of black bean peptide is extremely high and 85.02%, the amino acids are complete in variety and rich, and the ratio of essential amino acids to branched chain amino acids in the total content is high.

After the animal model and the drug model are successfully established, 50g of filler is taken and canned, the perfusion blood volume is 180mL, the flow rate is 100mL/min, the blood perfusion is continuous for 5d every week for 3 weeks, and after the perfusion is finished, the viscera of New Zealand rabbits are taken for measuring the immune index, and the measurement results are as follows:

experimental results:

1. referring to fig. 1, black bean peptide experimental group new zealand rabbits were dark in hair color, poor in gloss, and slow in feeding, after 5 days of continuous intraperitoneal injection of cyclophosphamide, while normal control group new zealand rabbits were glossy in coat color, and were normal in stool and urine and intake.

2. Referring to fig. 2, pathological changes in organ tissues of new zealand rabbits: the spleen tissue structure of the normal New Zealand rabbit is uniformly arranged, the spleen nodule is well developed, and the edges of the red marrow and the white marrow are imaged clearly under the mirror; the closely packed lymphocyte layer is observed under the mirror in thymus tissue, and the cortex and medulla are well-defined. Compared with a normal control group, the spleen nodule in the spleen of the New Zealand rabbit of the CY model group has a breakage phenomenon, and the red marrow and the white marrow at the center of the tissue are blurred in edge imaging; the boundary gap between the cortex and the medulla in the thymus tissue is increased, the hierarchy is unclear, the cortex area is reduced, and the medulla area is increased. Compared with CY model group, the spleen nodules of low, medium and high dose groups of black bean peptide are recovered; while the thymic cortex area increases, the cortical and medullary boundaries become progressively clearer and progressively closer to normal control levels.

3. Body weight and organ index change in New Zealand rabbits:

spleen and thymus are used as important immune organs of human body, play a certain role in coordination and protection in the process of cell growth, and can be widely involved in the process of nonspecific immunoregulation. Therefore, whether the medicine has influence on spleen index and thymus index can be used as a basic index of the scheme human immune mechanism.

TABLE 1 Effect of Black Soy bean peptide on New Zealand rabbit immune organ index

Note that: p <0.01# # compared to normal group p <0.05 p <0.01 compared to model group

The body weight and immune organ index analysis results of each group of New Zealand rabbits in this protocol are shown in Table 1. After 3 weeks of the experiment, there was no statistical difference in the final body weight (p > 0.05) for each group of New Zealand rabbits. The spleen index and thymus index of new zealand rabbits in the CY model group were increased compared to the normal control group, and the difference was extremely remarkable (p < 0.01); compared with the CY model group, the spleen index and thymus index of the black bean peptide low, medium and high dose groups are obviously reduced (p < 0.05), wherein the black bean peptide high dose group has extremely obvious difference (p < 0.01) compared with the spleen index of the New Zealand rabbit in the model group and gradually approaches to the normal level. In conclusion, the protein peptide can well regulate the immune organs of the organism after perfusion.

4. Changes in spleen lymphocyte proliferation effects in New Zealand rabbits:

activation of lymphocytes is the first means to defend against external invasion, and as a cellular immune transduction pathway, lymphocytes are finally in an activated state after division, proliferation and differentiation, are transformed into effector cells and perform various physiological functions. When T lymphocytes are stimulated, they divide into lymphoblasts, which increase cell volume and are metabolized. The extent of stimulation of ConA on lymphocytes can reflect the cellular immunity level of the body, and therefore can be used as one of the indicators for determining the immune function of the body.

The effect of spleen lymphoproliferation in each group of New Zealand rabbits in this protocol is shown in FIG. 3, and the lymphocyte stimulation index of the New Zealand rabbits is greatly reduced compared with that of the normal group, and the effect has extremely significant difference (p < 0.01), which indicates that the injection of cyclophosphamide can cause the reduced reactivity of the normal New Zealand rabbits to Canavalin A. After continuous blood perfusion for 3 weeks with black bean peptide, compared with the CY model group, the stimulation capability of lymphocytes of New Zealand rabbits (200-800 mg/kg) of black bean peptide dose group on the Canavalia gladiata protein is remarkably improved (p < 0.01), and the cell line is gradually close to a normal control group.

5. Changes in NK cell activity in new zealand rabbits:

NK cells in the organism are taken as a member of effector cells, have strong killing and clearing functions, and therefore have certain bactericidal and antiviral effects. In some cases, it also participates in hypersensitivity reactions and recognizes target cells to kill the medium. Can kill pathological cells in vitro without any antigen stimulation, and can participate in an immune regulation mechanism to a certain extent.

The effect of the NK cell activity of each group of New Zealand rabbits in this scheme is shown in FIG. 4, and the activity of the NK cells of the New Zealand rabbits is extremely remarkably reduced (p < 0.01) compared with that of the normal group in the model group, which indicates that the injection of cyclophosphamide can lead to the reduction of the activity of the NK cells of the normal New Zealand rabbits. After continuous blood perfusion for 3 weeks with black bean peptide, compared with the CY model group, the NK cell activity of New Zealand rabbits (200-800 mg/kg) of black bean peptide dose group is extremely obviously improved (p < 0.01), and gradually approaches to that of a normal control group.

6. Changes in macrophage phagocytic function in New Zealand rabbits:

the macrophage is used as a defensive cell with comprehensive functions, plays an important role in inherent immune reaction, and the activated macrophage can strengthen the function of phagocytic cells by secreting a large amount of carriers, thereby playing the roles of resisting viruses, tumors and the like. Therefore, the intensity of phagocytic function of macrophages can reflect the state of nonspecific immunity. Therefore, the evaluation of macrophage functions is mostly applied to the immune regulation effect of drugs on organisms.

The effect of phagocytic function of the new zealand rabbit macrophages in each group in this protocol is shown in figure 5, with the percentage and index of phagocytosis of the new zealand rabbit macrophages significantly reduced (p < 0.01) compared to the normal group. It was shown that cyclophosphamide injection resulted in a decrease in macrophage defence function in normal New Zealand rabbits. After continuous blood perfusion for 3 weeks through black bean peptide, compared with a CY model group, the black bean peptide dosage group can obviously improve the phagocytic capacity of the immunodeficient New Zealand rabbit macrophages, and can obviously improve the phagocytic percentage of the New Zealand rabbit macrophages (p < 0.05) when the dosage is 400 and 800 mg/kg; for the phagocytic index, the phagocytic index of New Zealand rabbits can be obviously improved (p < 0.05) at the dosage of 400 mg/kg.bw, and the phagocytic index of the New Zealand rabbits can be obviously improved (p < 0.01) by the black bean peptide at the dosage of 800mg/kg, and the novel Zealand rabbits are gradually close to the normal control group.

In view of the above, purification by hemodialysis by attaching a protein peptide to a resin is of great importance for effective alleviation of diseases caused by immunodeficiency and for complementary treatment.

Claims

1. The preparation method of the hemoperfusion filling material for treating immunodeficiency-induced tumors is characterized by comprising the following steps of mixing a black bean peptide solution and an inner core:

a. preparation of Black Soy peptide solution

secondly, freeze-drying the hydrolysate to obtain freeze-dried powder;

b. preparation of inner core

step two, mixing a black bean peptide solution and an inner core;

2. The method for preparing the hemoperfusion filler for treating immunodeficiency-induced tumors of claim 1, wherein in the first step, the biological glue is uniformly coated on the polystyrene resin by adopting an atomization spraying mode.

3. The method of claim 1 or 2, wherein in the first step, the black bean peptide is dissolved in a PBS buffer, and the PBS buffer is immersed in a polystyrene resin.

4. The method for preparing the hemoperfusion filler for treating immunodeficiency-induced tumors according to claim 1 or 2, wherein in the second step, a shaking table is adopted to fully and uniformly mix polystyrene resin with black bean peptide solution, and the shaking time is more than 2 hours.

5. The method for preparing the hemoperfusion filler for treating immunodeficiency-induced tumors according to claim 1 or 2, wherein in the second step, the polystyrene resin is poured into a dehydrator for drying.

6. The method for preparing the hemoperfusion filler for treating immunodeficiency-induced tumors according to claim 1 or 2, wherein in the first step, the black bean peptide solution is prepared into freeze-dried powder at a temperature of-50 to-80 ℃ by a freeze dryer.

7. The method for preparing the hemoperfusion filler for treating immunodeficiency-induced tumors according to claim 1 or 2, wherein the biological glue is n-octyl α -cyanoacrylate.