CN114146065A

CN114146065A - Chloroquine-coated denatured albumin nano-particle for selectively resisting inflammatory cells and preparation method and application thereof

Info

Publication number: CN114146065A
Application number: CN202111529666.2A
Authority: CN
Inventors: 查正宝; 马玉佩
Original assignee: Hefei University of Technology
Current assignee: Hefei University of Technology
Priority date: 2021-12-14
Filing date: 2021-12-14
Publication date: 2022-03-08

Abstract

The invention discloses a chloroquine-coated denatured albumin nano particle for selectively resisting inflammatory cells, and a preparation method and application thereof. The nano-particle has good stability and biocompatibility, can be internalized by macrophages with high efficiency, and can effectively aim at inflammatory cells at a colon part, so that inflammation is reduced to treat colitis diseases caused by dextran sodium sulfate.

Description

Chloroquine-coated denatured albumin nano-particle for selectively resisting inflammatory cells and preparation method and application thereof

Technical Field

The invention belongs to the technical field of nano materials, and particularly relates to chloroquine-coated denatured albumin nanoparticles for selectively resisting inflammatory cells.

Background

Inflammatory disease (IBD) is a disease of the digestive system and is roughly divided into two groups: ulcerative colitis and crohn's disease can affect the entire digestive tract, mainly ileum, colon and appendicitis. The pathogenesis and the cause of the disease are not clear at present, and the main symptoms of the disease are manifested by persistent diarrhea, weight loss, fatigue, hematochezia and abdominal pain. IBD is generally considered to be a chronic, relapsing disease caused by the interplay of multiple factors including genetic susceptibility, intestinal immune abnormalities, dysbacteriosis, and the environment. With the promotion of national economy and the gradual westernization of national life style, the number of cases of inflammatory bowel diseases in China is continuously increased in recent years. Although the number of death caused by IBD is few, the chronic and easily recurring characteristics of IBD can seriously affect the quality of life of patients, and moreover, the intestinal inflammation stimulation can increase the risk of patients suffering from complications such as thrombus, joint inflammation, primary cholangitis and the like, thereby causing serious economic stress and medical burden to families and society of patients. At present, no medicine capable of completely curing IBD exists in clinic, most of medicines for first-line treatment such as 5-aminosalicylic acid (5-ASA) are mainly used for relieving symptoms of patients but cannot be radically cured, and the patients are at risk of side effects such as nausea, vomiting, colitis symptom aggravation and hepatotoxicity after long-term administration. Therefore, the development of a safe and effective therapeutic strategy for IBD has important research value.

Since the first time in 1985 that japanese scholars prepared hamster ulcerative colitis models using Dextran Sulfate Sodium (DSS), there has been a large amount of data demonstrating that the causes, clinical symptoms, pathological changes and therapeutic responses of DSS colitis models are similar to human Ulcerative Colitis (UC). Therefore, the DSS colitis model becomes an important treatment means for researching UC etiology, pathogenesis and tumor diseases, and is one of the most widely applied UC models at present.

Chloroquine is widely applied to clinic as a low-price antimalarial drug at the earliest time, and the application in clinic is mainly used for treating autoimmune diseases such as rheumatoid arthritis, systemic lupus erythematosus and the like. In addition, chloroquine has been shown to have anticancer activity or prophylactic effect, and as a well-known autophagy inhibitor, p62 protein content can be accumulated. Surprisingly, more and more researches in recent years show that chloroquine also has strong anti-inflammatory capability and has good effect in a plurality of known inflammation models. However, in clinical practice, the toxic and side effects are large, and the administration time and dosage are large, so that the development of a carrier with excellent properties is urgently needed to carry and transport the carrier to the action site.

At present, for inflammatory diseases, nano-carriers are widely applied to carrying drugs to exert different effects, and different researchers and laboratories also develop various nano-particles for treatment. However, the traditional nanoparticles have the problems of high cost, difficult metabolism, difficult biological safety and the like, and the nanoparticles prepared by crosslinking after the desolvation of Bovine Serum Albumin (BSA) are a promising drug carrier. Bovine Serum Albumin (BSA) is a protein which is non-toxic, biodegradable, good in biocompatibility and low in price, and is widely applied to a drug sustained-release system. The denatured albumin nano-particles prepared by the ethanol desolventizing method still have good biocompatibility, and research reports show that the nano-particles can be efficiently internalized by inflammatory cells such as macrophages and neutrophils at inflammatory sites, so that the nano-particles have deep potential in the aspect of specific targeting. Therefore, the invention can be convenient for carrying chloroquine medicine to carry out specific delivery on inflammatory cells such as macrophage, neutrophil and the like at the colon part, so that the release of inflammatory factors is further reduced by inhibiting the accumulation of p62 protein caused by autophagy of the cells, and the aim of treating colitis is fulfilled.

Disclosure of Invention

In order to solve the problems of colitis diseases caused by dextran sodium sulfate, high cost, difficult metabolism, difficult guarantee of biological safety and the like of the traditional nanoparticles, the invention constructs the chloroquine-coated denatured albumin nanoparticles for selectively resisting inflammatory cells, and applies the chloroquine-coated denatured albumin nanoparticles to the treatment of the colitis diseases caused by dextran sodium sulfate DSS.

In order to solve the technical problem, the invention adopts the following technical scheme:

the invention firstly discloses a chloroquine-coated denatured albumin nano particle for selectively resisting inflammatory cells, which is characterized in that: the modified albumin nano particles wrapped with chloroquine are prepared by wrapping drug chloroquine with modified albumin nano particles serving as carriers, and the hydration particle size of the modified albumin nano particles is 50-150 nm.

The preparation method of the chloroquine-coated denatured albumin nano-particle comprises the following steps:

dissolving 80mg of bovine serum albumin in 4mL of deionized water to obtain a solution of 20mg/mL of bovine serum albumin; dissolving 16mg of chloroquine in 320 mu L of dimethyl sulfoxide to obtain a chloroquine solution with the concentration of 50 mg/mL; adding chloroquine solution into bovine serum albumin solution, and stirring at room temperature for reaction for 10-20 min; then dropwise adding 14mL of absolute ethyl alcohol, stirring and reacting at room temperature for 1h after dropwise adding, then adding 320 mu L of glutaraldehyde solution with the mass concentration of 2%, and reacting for 12-24h in a dark place: and (3) centrifuging the obtained product (the centrifugation temperature is 4 ℃) and washing the product with water to obtain the chloroquine-coated modified albumin nano particle.

The modified albumin nano particle coated with chloroquine has the function of remarkably relieving colitis caused by dextran sodium sulfate, has no remarkable toxicity to mammalian cells, has good biological safety, and can be used for preparing an effective anti-inflammatory preparation for resisting colitis caused by dextran sodium sulfate.

The mechanism for selectively resisting inflammatory cells in a colon part by the chloroquine-coated denatured albumin nano-particles is as follows: in a DSS-induced inflammatory bowel disease model, inflammatory cells such as macrophages and neutrophils are actively recruited to a colon part to resist the generation of situations such as the increase of the level of inflammatory factors, and the synthesized chloroquine-coated denatured albumin nanoparticles can be effectively internalized by the macrophages and the neutrophils, so that the synthesized nanoparticles can reach the inflammatory part more efficiently. When the denatured albumin wraps chloroquine to reach an inflammation part, on one hand, the chloroquine drug can reduce the level of inflammatory factors and inhibit the generation of inflammatory bodies so as to slow down inflammation, thereby achieving the result of relieving treatment. On the other hand, the chloroquine drug, as a well-known autophagy inhibitor, can also achieve the effect of treating inflammation by inhibiting macrophage autophagy in the colon part to increase the content of P62 protein so as to inhibit the macrophages from secreting inflammatory factors. In conclusion, chloroquine is a cheap and effective drug for treating colitis, and the modified albumin nanoparticles are a good and safe carrier system.

The invention has the beneficial effects that:

1. the modified albumin nano particle wrapped by chloroquine has good stability and biocompatibility. Chloroquine is a cheap and effective medicament for treating colitis, and the modified albumin nano particles are a good and safe carrier system, so that the safety of clinical application is improved; meanwhile, the macrophage and the neutrophil at the inflammation part efficiently take up the denatured albumin nano particles, so that the efficient enrichment of the material at the inflammation part is realized.

2. The modified albumin nano particle coated with chloroquine is used for resisting DSS induced colitis, the anti-inflammatory performance of the traditional chloroquine medicine is obviously improved, and the medicine concentration and dosage are obviously reduced.

3. The modified albumin nano particle coated with chloroquine has the advantages of simple preparation process, mild conditions, possibility of large-scale production and potential for industrial and practical application.

4. The material used in the invention has good biocompatibility, no direct or indirect toxic action on human body and no potential toxicity.

5. The modified albumin nano particle wrapped by chloroquine has good dispersibility and stability, and is beneficial to clinical use.

Drawings

FIG. 1 is a transmission electron micrograph of chloroquine-coated denatured albumin nanoparticles prepared in example 1.

FIG. 2 is a scanning electron micrograph of chloroquine-coated denatured albumin nanoparticles prepared in example 1.

Fig. 3 is a hydrated particle size diagram of chloroquine-coated denatured albumin nanoparticles prepared in example 1.

Fig. 4 is an ultraviolet-visible absorption spectrum of the chloroquine-coated modified albumin nanoparticles, the chloroquine-free modified albumin nanoparticles and chloroquine prepared in example 1.

Fig. 5 shows the encapsulation efficiency of chloroquine-coated denatured albumin nanoparticles prepared in example 1 with different mass ratios.

Fig. 6 is a drug release profile of chloroquine-coated denatured albumin nanoparticles prepared in example 1.

Fig. 7 is a 7-day stability graph of chloroquine-encapsulated denatured albumin nanoparticles prepared in example 1 in different solvents.

FIG. 8 is a graph showing the in vitro hemolytic performance test of the chloroquine-coated denatured albumin nanoparticles prepared in example 1.

Fig. 9 is a schematic flow chart for constructing a DSS-induced colitis model.

FIG. 10 is a graph showing the change of body weight of Balb/c female mice after DSS treatment and water treatment, in comparison with chloroquine-coated denatured albumin nanoparticles, non-chloroquine-coated denatured albumin nanoparticles and chloroquine prepared in example 1.

FIG. 11 is a graph showing the change of disease activity index of Balb/c female mice treated with DSS and water, in comparison with chloroquine-coated modified albumin nanoparticles, non-chloroquine-coated modified albumin nanoparticles and chloroquine prepared in example 1.

FIG. 12 is a photograph of colon of Balb/c female mice treated with DSS and water, showing modified albumin nanoparticles coated with chloroquine prepared in example 1, and modified albumin nanoparticles not coated with chloroquine and chloroquine.

FIG. 13 is a graph of colon length of Balb/c female mice treated with DSS and water treated with chloroquine, and modified albumin nanoparticles coated with chloroquine, and modified albumin nanoparticles not coated with chloroquine prepared in example 1.

FIG. 14 is a colon hematoxylin-eosin stained section of Balb/c female mice treated with DSS and water, prepared in example 1, with modified albumin nanoparticles coated with chloroquine, modified albumin nanoparticles not coated with chloroquine, and chloroquine.

FIG. 15 is a chart of IL-1 β immunohistochemical staining of colon of Balb/c female mice treated with DSS and water, with chloroquine-coated denatured albumin nanoparticles prepared in example 1, and with denatured albumin nanoparticles not coated with chloroquine and chloroquine.

FIG. 16 is a chart of immunohistochemical staining of colon IL-6 of Balb/c female mice after DSS treatment and water treatment with chloroquine-coated denatured albumin nanoparticles prepared in example 1, and with chloroquine-non-coated denatured albumin nanoparticles and chloroquine.

FIG. 17 is a chart of TNF-alpha immunohistochemical staining of colon of Balb/c female mice after DSS treatment and water treatment with chloroquine-coated denatured albumin nanoparticles prepared in example 1, and with chloroquine-non-coated denatured albumin nanoparticles and chloroquine.

Detailed Description

In order to make the aforementioned objects, features and advantages of the present invention more comprehensible, specific embodiments thereof will be described in detail with reference to the following examples. The following is merely exemplary and illustrative of the inventive concept and various modifications, additions and substitutions of similar embodiments may be made to the described embodiments by those skilled in the art without departing from the inventive concept or exceeding the scope of the claims defined thereby.

Example 1

In this example, chloroquine-coated denatured albumin nanoparticles were prepared as follows:

dissolving 80mg of bovine serum albumin in 4mL of deionized water to obtain a solution of 20mg/mL of bovine serum albumin; dissolving 16mg of chloroquine in 320 mu L of dimethyl sulfoxide to obtain a chloroquine solution with the concentration of 50 mg/mL; adding chloroquine solution into bovine serum albumin solution, and then stirring and reacting for 15min at room temperature at a stirring speed of 1600 rpm; then, 14mL of absolute ethyl alcohol is dropwise added, stirring reaction is carried out at the stirring speed of 600rpm for 1h at room temperature after the dropwise addition is finished, 320 mu L of glutaraldehyde solution with the mass concentration of 2% is added, and light-shielding reaction is carried out for 12 h: and centrifuging the obtained product (the centrifugation temperature is 4 ℃, and the rotation speed is 18000-.

Fig. 1 and fig. 2 are a transmission electron microscope image and a scanning electron microscope image of the chloroquine-coated modified albumin nanoparticle prepared in this example, respectively, and it can be seen from the images that the nanoparticle is in a spherical state and has a diameter of 50-150 nm.

Fig. 3 is a hydrated particle size diagram of chloroquine-coated denatured albumin nanoparticles prepared in this example, from which it can be seen that: the hydrated particle size of the nano particles is about 85nm, the dispersibility is good, and the particle sizes of most nano particles are concentrated between 50nm and 150 nm.

Fig. 4 is a graph showing the uv-vis absorption spectra of Chloroquine-coated modified albumin nanoparticles (BSA-Chloroquine), Chloroquine-free modified albumin-coated nanoparticles (BSA), and Chloroquine (Chloroquine) prepared in this example. The characterization method comprises the following steps: three sample solutions were prepared: the BSA content of the modified albumin nano particles wrapping chloroquine is 20mg/mL, and the equivalent of chloroquine is 400 mug/mL; the BSA content of the modified albumin nano particles not wrapped by chloroquine is 20 mg/mL; the content of free chloroquine is 400 mug/mL; 3mL of each sample was added to the UV cuvette and the UV-VIS absorption spectrum was measured. From the figure, it can be seen that the modified albumin nanoparticles encapsulating chloroquine show obvious absorption peaks at 300-400nm, which coincide with the positions of the absorption peaks appearing at 300-400nm of single chloroquine, and confirm that chloroquine is successfully encapsulated by the modified albumin nanoparticles.

FIG. 5 shows the encapsulation efficiency of modified chloroquine-encapsulated albumin nanoparticles prepared according to different mass ratios of BSA and chloroquine in this example. The characterization method comprises the following steps: preparing solutions of the nano particles with different mass ratios and solutions of the modified albumin nano particles without coating chloroquine (the BSA content is 20mg/mL), taking 3mL respectively, testing an ultraviolet-visible absorption spectrogram of each solution, taking the nano particle solution without coating chloroquine as a base line, and substituting an absorption value at 340nm into a standard curve of the chloroquine to obtain the encapsulation efficiency of the nano particles with different mass ratios. As can be seen from the figure, the mass ratio of bovine serum albumin to chloroquine is 5: the best encapsulation efficiency is obtained when the molecular weight is 1.

Fig. 6 is a drug release curve chart of the chloroquine-coated denatured albumin nanoparticle prepared in this example, and its characterization method is as follows: 2mL of Chloroquine-coated denatured albumin nanoparticles (BSA-Chl) with a Chloroquine equivalent of 400 μ g/mL and a free Chloroquine (Chloroquine) (400 μ g/mL) aqueous dispersion are added into a dialysis bag with a molecular weight of 1000MW, 30mL of PBS buffer solution with a pH value of 7.4 or 5 (buffer solution with a pH value of 7.4 is added at the periphery of the free Chloroquine dialysis bag) is added at the periphery of the dialysis bag to be incubated at a shaking table at 37 ℃, ultraviolet-visible light absorption spectra are tested at a plurality of time points, and an absorption value at 340nm is substituted into a Chloroquine concentration standard curve for quantification. It can be seen from the figure that chloroquine coated by denatured albumin has a slow release effect, and the maximum drug release amount is increased under the PBS medium with the pH value of 5.

Fig. 7 is a 7-day stability chart of chloroquine-coated denatured albumin nanoparticles prepared in this example in different media (water, PBS buffer solution with pH of 7.4 and DMEM in high-glucose medium), which is characterized by the following steps: dispersing the prepared modified albumin nano particles coated with chloroquine in different media, storing at normal temperature, and testing the sizes of the modified albumin nano particles by using a laser particle sizer every other day. It can be seen from the figure that the prepared nanoparticles have good stability, and keep stable under the condition of water dispersion, and the size of the nanoparticles is basically unchanged.

Fig. 8 is a graph of in vitro hemolytic performance test of chloroquine-coated denatured albumin nanoparticles prepared in this example, which is characterized by the following steps: diluting the nanoparticle aqueous dispersion to 25 mug/mL, 50 mug/mL, 100 mug/mL, 200 mug/mL, 400 mug/mL and 800 mug/mL chloroquine equivalent, adding 4.5mL physiological saline into 500 mug fresh blood for centrifugal washing for 5-8 times, wherein the centrifugal speed is 3000rpm and the centrifugal time is 10min, after the blood supernatant is clear and transparent, discarding the supernatant, using the physiological saline to fix the volume to 5mL), mixing the solution with 0.2mL treated blood (500 mug fresh blood), incubating for 4h at 37 ℃, then centrifuging for 10min at 3000rpm, and absorbing the absorbance value of the supernatant at OD541 nm to calculate the hemolysis rate. It can be seen from the figure that the hemolysis rate of the nanoparticles with different concentrations is lower than 5%, indicating that the biocompatibility of the material is good.

To verify the therapeutic effect of the chloroquine-coated denatured albumin nanoparticles prepared in this example on DSS-induced colitis, the following DSS model was established: 50 Balb/c female mice were divided into five groups: normal control group PBS + Water: the cells were intravenously injected with a PBS buffer solution of pH 7.4, and were incubated with water throughout. (ii) DSS control group PBS + DSS: the cells were intravenously injected with a PBS buffer solution having a pH of 7.4, and were incubated with a 3% DSS aqueous solution for one week after acclimation. ③ blank vector control group BSANPs + DSS: the dispersions of BSANPs in PBS (PBS buffer pH 7.4) were injected intravenously and incubated with 3% by mass DSS in water for one week after acclimation. (iv) nanoparticle treatment group BSA-chl NPs + DSS: PBS dispersion of BSA-chl NPs (PBS buffer solution with pH 7.4) was injected intravenously, and was incubated with 3% mass concentration DSS aqueous solution for one week after acclimation. Free drug control Free chl + DSS: a PBS dispersion (PBS buffer solution with pH 7.4) of an intravenous chloroquine drug was incubated with 3% DSS aqueous solution at a mass concentration for one week after the adaptation. Ten of the above groups are placed. And (3) carrying out tail vein injection administration on the modified albumin nano particles coated with chloroquine, the modified albumin nano particles not coated with chloroquine and chloroquine on

days

7, 9, 11 and 13 established in a DSS model.

100. mu.L of a PBS dispersion of chloroquine-coated denatured albumin nanoparticles (chloroquine concentration equivalent in nanoparticles is 5mg/mL, and this concentration is 25mg of chloroquine-equivalent nanoparticles injected in vivo per kg of mouse body weight), 100. mu.L of a PBS dispersion of free chloroquine (free chloroquine concentration is 10mg/mL, and this concentration is 50mg of free chloroquine injected in vivo per kg of mouse body weight), and 100. mu.L of a PBS dispersion of denatured albumin nanoparticles not coated with chloroquine (BSA concentration is consistent with the above-mentioned concentration of the coated drug) were administered on

days

7, 9, 11, and 13 of the DSS model set-up.

The growth of the mice in each experimental group was observed by the change in the weight percentage of the mice in each group, and the results are shown in fig. 10. It can be seen that the body weight of normal mice increases continuously within 16 days, while the PBS + DSS treated mice have significant weight loss, and the treatment with chloroquine-coated denatured albumin nanoparticles increased the body weight of the mice, indicating that the treatment with chloroquine-coated denatured albumin nanoparticles can significantly alleviate typical inflammatory symptoms.

The growth of mice in each experimental group was observed by the change of disease activity index of mice in each group. As shown in fig. 11, it can be seen that the body weight of normal mice has almost no change in DAI score within 16 days, while PBS + DSS treated mice have a significant increase in disease activity index, and chloroquine-coated denatured albumin nanoparticles decrease the disease activity index of mice, indicating that treatment with chloroquine-coated denatured albumin nanoparticles can significantly alleviate typical inflammatory symptoms.

On day 16, each group of mice was dissected and colons were isolated by taking a visual picture of the colons of each group. The results are shown in fig. 12, and it can be seen that the shortening of the colon in the PBS + DSS group and DSS + BSANPs restored the length of the colon to normal levels after treatment with chloroquine-coated denatured albumin nanoparticles.

Each group of mice was dissected and colons were isolated on day 16, and a colon length comparison chart was observed for each group by measuring the colons of each group. As shown in FIG. 13, it can be seen that the colon was shortened from 9.2+0.2cm to 7.6+0.2cm in the PBS + DSS group and DSS + BSANPs, and the length of the colon was restored to a normal level after the treatment with chloroquine-coated denatured albumin nanoparticles, as compared to healthy mice.

To further demonstrate the therapeutic effect of the chloroquine-coated denatured albumin nanoparticles obtained in this example on DSS-induced colitis, the following tests were performed: on day 16, each group of mice was dissected and colons were isolated, and the isolated colon tissue pieces (generally no more than 0.5 cm thick) were placed in a pre-prepared fixative (10% formalin, Bouin's fixative) to denature and coagulate the proteins of the tissue, thereby preventing autolysis after cell death and maintaining the original morphological structure of the cells.

The H & E sections were prepared after dehydration and clearing, wax immersion and embedding, slicing and pasting, dewaxing, hematoxylin-eosin staining, dehydration and clearing, sealing and the like, and as a result, as shown in fig. 14, it can be seen that significant destruction of colon microstructure and disruption of goblet cells can be observed in colitis mice (PBS + DSS group and DSS + BSANPs group).

The IL-1 beta immunohistochemical section is prepared by the steps of decoloring, washing and soaking, antibody application, color development, full washing, counterstaining, dehydration, transparence, mounting and the like, and the result is shown in figure 15, so that the level of inflammatory factors in colitis mice (PBS + DSS group and DSS + BSANPs group) is obviously improved and is obviously higher than that of a normal group and a treatment group, which indicates that the section has better treatment capability in a DSS-induced colitis model.

The IL-6 immunohistochemical section is prepared by the steps of decoloring, washing and soaking, antibody coating, color development, full washing, counterstaining, dehydration, transparence, mounting and the like, and the result is shown in figure 16, so that the level of inflammatory factors in colitis mice (PBS + DSS group and DSS + BSA NPs group) is obviously improved and is obviously higher than that of a normal group and a treatment group, which indicates that the section has better treatment capability in a DSS-induced colitis model.

The TNF-alpha immunohistochemical section is prepared by the steps of decoloring, washing and soaking, antibody application, color development, full washing, counterstaining, dehydration, transparence, mounting and the like, and the result is shown in figure 17, so that the level of inflammatory factors in colitis mice (PBS + DSS group and DSS + BSANPs group) is obviously improved and is higher than that of a normal group and a treatment group, which indicates that the TNF-alpha immunohistochemical section has better treatment capability in a DSS-induced colitis model.

The present invention is not limited to the above exemplary embodiments, and any modifications, equivalent replacements, and improvements made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A chloroquine-encapsulated denatured albumin nanoparticle for selective combating inflammatory cells, characterized by: the modified albumin nano particle wrapped by chloroquine takes the modified albumin nano particle as a carrier and is wrapped by chloroquine.

2. The chloroquine-encapsulated denatured albumin nanoparticle of claim 1, wherein: the hydration particle size of the modified albumin nano particles wrapping chloroquine is 50-150 nm.

3. A method for preparing chloroquine-coated denatured albumin nanoparticles according to claim 1 or 2, characterized in that:

dissolving bovine serum albumin in deionized water to obtain a bovine serum albumin solution; dissolving chloroquine in dimethyl sulfoxide to obtain a chloroquine solution; adding chloroquine solution into bovine serum albumin solution, and stirring at room temperature for reaction for 10-20 min; and then dropwise adding absolute ethyl alcohol, stirring and reacting at room temperature for 1h after dropwise adding, then adding a glutaraldehyde solution with the mass concentration of 2%, and reacting for 12-24h in a dark place: and centrifuging and washing the obtained product to obtain the chloroquine-coated denatured albumin nano particles.

4. The production method according to claim 3, characterized in that: the concentration of the bovine serum albumin solution is 20 mg/mL.

5. The production method according to claim 3, characterized in that: the concentration of the chloroquine solution is 50 mg/mL.

6. The production method according to claim 3, characterized in that: the mass ratio of the bovine serum albumin to the chloroquine is 5: 1.

7. The production method according to claim 3, characterized in that: the volume ratio of the absolute ethyl alcohol to the bovine serum albumin is 14 mL: 80 mg.

8. The production method according to claim 3, characterized in that: the ratio of the volume of the glutaraldehyde solution to the mass of the bovine serum albumin is 320 μ L: 80 mg.

9. The production method according to claim 3, characterized in that: the temperature of the centrifugation was 4 ℃.

10. The use of the chloroquine-coated denatured albumin nanoparticles of any one of claims 1-2, wherein: for the preparation of an anti-inflammatory agent against colitis caused by dextran sodium sulphate.