CN117482067A - Nanoparticle with neutral granulocyte membrane wrapping PIM2 kinase small molecule inhibitor and preparation method and application thereof - Google Patents
Nanoparticle with neutral granulocyte membrane wrapping PIM2 kinase small molecule inhibitor and preparation method and application thereof Download PDFInfo
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
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- A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
- A61K9/51—Nanocapsules; Nanoparticles
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- A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
- A61K31/445—Non condensed piperidines, e.g. piperocaine
- A61K31/4523—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems
- A61K31/454—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems containing a five-membered ring with nitrogen as a ring hetero atom, e.g. pimozide, domperidone
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- A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
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Abstract
The invention belongs to the technical field of medical biology, and particularly discloses a nanoparticle of a PIM2 kinase small molecule inhibitor wrapped by a neutrophil membrane, a preparation method and application thereof, wherein neutrophils are obtained and stored under liquid nitrogen for subsequent membrane separation; separating neutrophil granulocyte membrane from neutrophil granulocyte by repeated freeze thawing method; synthesizing a nanoparticle with a neutral granulocyte membrane wrapping the PIM2 kinase small molecule inhibitor. Compared with the prior art, the nanoparticle prepared by wrapping the PIM2 kinase small molecule inhibitor AZD1208 by using the neutral granulocyte membrane can target and inhibit PIM2 kinase to reverse macrophage M1 polarization, strengthen and relieve autoimmune arthritis, and has better targeting and better treatment effects and lower drug side effects compared with the treatment of autoimmune arthritis by using AZD1208 alone.
Description
Technical Field
The invention relates to the technical field of medical biology, in particular to a nanoparticle of a PIM2 kinase small molecule inhibitor wrapped by a neutral granulocyte membrane, a preparation method and application thereof.
Background
Autoimmune arthritis is an arthritis in which the body attacks the joints and connective tissues due to the abnormality of the immune system, and is a local manifestation of systemic autoimmune diseases at joint sites, including ankylosing spondylitis, rheumatoid arthritis, and the like. Autoimmune arthritis can accumulate at various joint sites throughout the body, early stage of joint swelling, pain, stiffness, and gradually develop into late stage of rigidity and deformity. The pathogenesis of autoimmune arthritis is not clear, and it is considered that the pathogenesis of autoimmune arthritis is the result of the combined action of various factors, such as immune factors, genetic factors, intestinal dysbacteriosis, infectious factors, environmental factors and the like. Recent studies have shown that proinflammatory macrophages may dominate autoimmune arthritis. Macrophages are largely classified into two types, pro-inflammatory (M1) and anti-inflammatory (M2). It has been demonstrated that the deregulation of M1 type macrophages plays an important role in the pathogenesis of a variety of autoimmune arthritis such as rheumatoid arthritis, ankylosing spondylitis, psoriatic arthritis and the like.
Post-translational modifications (posttranslation modification, PTM) refer to post-translational chemical modifications of proteins that reversibly modulate the activity and function of the protein to accommodate changes in different cellular states and environments. PTM has been found to be closely associated with a variety of autoimmune diseases including rheumatoid arthritis, systemic lupus erythematosus, and the like. PIM kinases are a class of serine/threonine kinases that include three isozymes PIM-1, PIM2, PIM-3. It is involved in posttranslational modification of various protein substrates, and plays an important role in apoptosis and proliferation, cell cycle regulation, migration, and energy metabolism. Recent studies have shown that PIM2 can enhance the proliferative, anti-apoptotic and invasive metastatic capacity of tumor cells by mediating phosphorylation of a variety of glycolytic key enzymes, promoting aerobic glycolysis. Meanwhile, PIM2 is also involved in regulating the development and function of immune cells. It was found that cells significantly up-regulate PIM2 in an inflammatory environment, and that up-regulation of PIM2 further promotes expression of inflammatory cytokines such as IL-6 and IL-1β.
The current clinical medicines for treating autoimmune arthritis mainly comprise nonsteroidal anti-inflammatory drugs (NSAIDs), tumor Necrosis Factor Inhibitors (TNFi) and disease-relieving antirheumatic drugs (DMARDs). NSAIDs can improve symptoms of lumbago, backache, morning stiffness and joint pain of partial patients, but still has poor treatment response of partial patients to NSAIDs, and has obvious side effects after long-term administration. The DMARDs are mainly used for patients with peripheral arthritis, and have poor treatment effect on AS mainly caused by spinal lesions. TNFi can block TNF activity and inhibit lesion progression, and is suitable for patients with non-effective and extra-articular symptoms due to NSAIDs. Therefore, a new target for treating autoimmune arthritis needs to be explored, a small molecular inhibitor for specifically inhibiting the target is sought, and the neutral granulocyte membrane is used for wrapping the drug-loaded nano particles, so that the targeting property of the drug-loaded nano particles is further improved, and further, the targeted therapeutic effect of the drug-loaded nano particles in treating autoimmune arthritis is explored, so that a safe and effective new method is brought to clinical treatment of autoimmune arthritis.
Disclosure of Invention
In order to solve the technical problems, the invention provides a nanoparticle with a neutral particle cell membrane wrapping PIM2 kinase small molecule inhibitor, and a preparation method and application thereof.
In order to achieve the above purpose, the invention is implemented according to the following technical scheme:
the first object of the present invention is to provide a method for preparing a nanoparticle of a small molecular inhibitor of PIM2 kinase encapsulated by a neutrophil membrane, comprising the steps of:
s1, obtaining neutrophils and preserving the neutrophils under liquid nitrogen for subsequent membrane separation;
s2, separating neutrophil granulocyte membranes from the neutrophil granulocytes by using a repeated freeze thawing method;
s3, synthesizing nanoparticles of a PIM2 kinase small molecule inhibitor wrapped by a neutrophil membrane:
s31, dissolving 2.5mg of PIM2 kinase small molecule inhibitor AZD1208 in 1ml of dimethyl sulfoxide DMSO to obtain solution A, dissolving 10mg of MMP2 polypeptide in 9ml of water to obtain solution B, mixing the solution A and the solution B, performing ultrasonic treatment on ice in an ultrasonic breaker with the power of 50% for 30min to obtain solution C, dropwise adding the solution C into 10ml of water, and stirring at 4 ℃ for 8-12h to obtain solution D after the dropwise adding is completed within 2 min;
s32, mixing the neutrophil membrane with the solution D according to the volume ratio of the neutrophil membrane to the solution D=1:2.5, performing ultrasonic treatment for 30min, stirring for 2h, extruding liquid by a cell membrane extruder or filtering the liquid by a 0.8 mu m cell filter after extruding by a syringe to obtain the nano-particles of the neutrophil membrane-encapsulated PIM2 kinase small molecular inhibitor.
Further, the step S1 specifically includes:
intraperitoneal injection of 1.5mg/kg lipopolysaccharide LPS in mice to activate neutrophils; after 6h, peripheral blood of the mice is collected in a test tube containing an anticoagulant EDTA-2K, and neutrophils are separated by using a peripheral blood neutrophil separation kit of the mice; the isolated neutrophils were then resuspended in 1M sucrose solution to give neutrophils and stored under liquid nitrogen.
Further, the step S2 specifically includes:
re-suspending the neutrophils with 1mL pre-chilled double distilled water, then sonicating the mixture at 42kHz for 10min, and performing 3 freeze-thawing cycles; freezing at-80deg.C for 30min, and thawing at room temperature for 30min; centrifuging at 4deg.C under 10000g centrifugal force for 10min, collecting supernatant, and centrifuging at 120000g centrifugal force for 45min; after centrifugation, the neutrophil membrane is obtained and stored at 80 ℃ for standby.
Preferably, 20mg of DSPE-PEG-COOH is added after mixing the solution A and the solution B in the step S31.
A second object of the present invention is to provide a nanoparticle having a small molecular inhibitor of PIM2 kinase encapsulated by a neutrophil membrane prepared by the above method.
The third object of the invention is to provide an application of the nanoparticle of the PIM2 kinase small molecular inhibitor wrapped by the neutral granulocyte membrane in preparing a medicament for preventing and treating autoimmune arthritis, wherein the medicament for preventing and treating autoimmune arthritis targets and inhibits macrophage M1 polarization for the nanoparticle of the PIM2 kinase small molecular inhibitor wrapped by the neutral granulocyte membrane.
Preferably, the medicament further comprises a pharmaceutically acceptable carrier and/or adjuvant.
Preferably, the dosage forms of the medicine comprise injection, oral liquid, tablet, granule, capsule and pill.
Compared with the prior art, the nanoparticle prepared by wrapping the PIM2 kinase small molecule inhibitor AZD1208 by using the neutral granulocyte membrane can target and inhibit PIM2 kinase to reverse macrophage M1 polarization, strengthen and relieve autoimmune arthritis, and has better targeting and better treatment effects and lower drug side effects compared with the treatment of autoimmune arthritis by using AZD1208 alone.
Drawings
FIG. 1 is a schematic representation of the synthesis of nanoparticles of the present invention wherein the neutrophil membrane encapsulates a PIM2 kinase small molecule inhibitor.
FIG. 2 is a transmission electron microscope appearance, zeta potential and drug release profile of nanoparticles with neutrophil membrane encapsulating PIM2 kinase small molecule inhibitors.
FIG. 3 shows PIM2 expression in Bone Marrow Derived Macrophages (BMDM) of mice during M1 polarization.
Fig. 4 is a graph showing that the PIM2 kinase small molecule inhibitor AZD1208 inhibits the M1 polarization of BMDM.
Fig. 5 is a graph showing CIA mouse arthritis scores after treatment with the PIM2 kinase small molecule inhibitor AZD 1208.
FIG. 6 shows the polarization of the nanoparticles of the small molecular inhibitor of PIM2 kinase encapsulated by the neutrophil membrane in the action of MMP2 on macrophage M1.
FIG. 7 shows the arthritis score of CIA mice treated with nanoparticles of a small molecule inhibitor of neutrophil membrane-encapsulated PIM2 kinase.
FIG. 8 shows the toxic side effects of nanoparticles of a small molecular inhibitor of PIM2 kinase encapsulated by neutrophil membrane on various organs of mice.
Detailed Description
The present invention will be described in further detail with reference to the following examples in order to make the objects, technical solutions and advantages of the present invention more apparent. The specific embodiments described herein are for purposes of illustration only and are not intended to limit the invention.
The experimental methods in the following examples, unless otherwise specified, are conventional, and the experimental materials used in the following examples, unless otherwise specified, are commercially available.
Example 1
The preparation of the nanoparticle with the PIM2 kinase small molecule inhibitor wrapped by the neutral granulocyte membrane is shown in figure 1, and comprises the following specific steps:
(1) Neutrophils were obtained and stored under liquid nitrogen for subsequent membrane separation:
intraperitoneal injection of 1.5mg/kg lipopolysaccharide LPS in mice to activate neutrophils; after 6h, peripheral blood of the mice is collected in a test tube containing an anticoagulant EDTA-2K, and neutrophils are separated by using a peripheral blood neutrophil separation kit of the mice; the isolated neutrophils were then resuspended in 1M sucrose solution to give neutrophils and stored under liquid nitrogen.
(2) Separating neutrophil granulocyte membrane from neutrophil by repeated freeze thawing method:
re-suspending the neutrophils with 1mL pre-chilled double distilled water, then sonicating the mixture at 42kHz for 10min, and performing 3 freeze-thawing cycles; freezing at-80deg.C for 30min, and thawing at room temperature for 30min; centrifuging at 4deg.C under 10000g centrifugal force for 10min, collecting supernatant, and centrifuging at 120000g centrifugal force for 45min; after centrifugation, the neutrophil membrane is obtained and stored at 80 ℃ for standby.
(3) Synthesis of nanoparticles with neutrophil membrane encapsulation of PIM2 kinase small molecule inhibitors:
dissolving 2.5mg of PIM2 kinase small molecule inhibitor AZD1208 in 1ml of dimethyl sulfoxide DMSO to obtain solution A, dissolving 10mg of MMP2 polypeptide in 9ml of water to obtain solution B, mixing the solution A and the solution B, adding 20mg of DSPE-PEG-COOH, performing ultrasonic treatment on ice in a 50% power ultrasonic breaker for 30min to obtain solution C, dripping the solution C into 10ml of water, and stirring at 4 ℃ for 8-12h to obtain solution D after the dripping is completed within 2 min; mixing the neutrophil membrane with the solution D according to the volume ratio of the neutrophil membrane to the solution D=1:2.5, performing ultrasonic treatment for 30min, stirring for 2h, extruding liquid by a cell membrane extruder or filtering the liquid by a 0.8 mu M cell filter after extrusion by a syringe to obtain the nano-particles (NP@NM-M) of the small-molecule inhibitor of the PIM2 kinase, wherein the nano-particles are wrapped by the neutrophil membrane.
The morphology of the neutrophil-encapsulated AZD 1208-loaded nanoparticle (NP@NM-M) was examined by a transmission electron microscope with the zeta potential measured by a Mo Erwen particle size analyzer, and the results are shown in FIG. 2, and as can be seen from FIG. 2: the NP@NM-M has good synthesis effect and can form round-like nano particles; under the action of MMP2, NP@NM-M has better drug release capability.
Example 2
The PIM2 kinase small molecule inhibitor has an effect of preventing and treating autoimmune arthritis by inhibiting M1 polarization of BMDM, and the specific process is as follows:
1. in vitro test
(1) BMDM separation
After dislocation of C57BL/J mice, the mice were rinsed with 75% alcohol for 5min. The tibia and femur of the mice were removed in a sterile environment and the epiphyses at both ends of the bone were cut to facilitate needle insertion. Serum-free culture medium DMEM was repeatedly added to the bone marrow cavity until the bone cavity became white. Bone marrow cells were centrifuged at 1350rpm for 10min at room temperature. After discarding the supernatant, red blood cell lysate was added and incubated for 3min. DMEM was added to stop the reaction, centrifuged and resuspended in the appropriate medium. The suspension was filtered using a 70 μm sterile cell filter. After centrifugation, the cells were cultured in DMEM containing 10% fetal calf serum, 100U/mL penicillin streptomycin, 50. Mu.M beta. -mercaptoethanol and 20ng/mL mouse macrophage colony stimulating factor.
(2) Induction of BMDM differentiation
After isolation of BMDM, the cell concentration was adjusted to 2X 10 6 Plating/ml on a twelve-well plate, adding 20ng/ml M-CSF to the complete medium to induce maturation into macrophages; about 5-7 days, differentiating and maturing into macrophages (adherence), and carrying out medium replacement (or half-quantity liquid replacement) in the middle of 3 days, wherein F4/80+CD11b+ double positive expression can be selected as a marker protein for identifying macrophage maturation; mature macrophages were digested and re-plated to induce polarization towards M1: m1 polarization factor: LPS (100 ng/ml) +IFN-. Gamma.s (50 ng/ml); after 24 hours of stimulation, the M1, M1 macrophages up-regulate expression of CD86, MHC-II and the like after polarization are detected by a flow method; induction of polarization of M2: m2 polarization factor: IL-4 (10 ng/ml) +IL-10 (or IL-13) (10 ng/ml); m2 macrophages up-regulate expression of CD206, and the like.
(3) Proteins extracted from cell lysates were separated by 10% SDS-PAGE and then transferred to a polyvinylidene fluoride (PVDF) membrane. After blocking with 5% nonfat milk powder for 1.5 hours, the proteins were incubated with anti-beta-actin (1:1000), PIM2 (1:1000) overnight at 4 ℃. PVDF membranes were washed 3 times with TBST bufer and incubated with goat anti-rabbit IgG H & L (HRP) secondary antibody (1:5000) for 1 hour at room temperature. After washing with TBST, protein detection was performed using ECL reagent and ChemiScope 6100 chemiluminescent imaging system. The strips were subjected to at least 3 quantitative analyses. In immunoblotting experiments, beta-actin was used as a control for other proteins. The results are shown in fig. 3, and as can be seen from fig. 3: mouse Bone Marrow Derived Macrophages (BMDM) significantly up-regulate PIM2 expression during M1 polarization.
To assess the polarization of macrophage M1, BMDM cells were plated at 2X 10 per well 5 The individual cells were seeded at a rate of 12-well plate and, after differentiation into mature macrophages by the method described above,different stimuli were given. M0 group was used as control group without any stimulation; LPS (100 ng/mL) was added to group M1; LPS (100 ng/mL) +AZD1208 (20 uM) was added to the M1+AZD group. After 24h of stimulation, the samples were stained with PE anti-mouse MHC-II antibody and PI anti-mouse F4/80 antibody for 30min and detected using a flow cytometer (thermofiser), and the data were analyzed using FlowJo software, as shown in fig. 4, and as can be seen from fig. 4: the PIM2 kinase small molecule inhibitor AZD1208 is capable of inhibiting M1 polarization of BMDM.
2. In vivo test
Establishment of CIA arthritis mouse model: DBA mice (8-12 weeks old) were produced by Jiangsu Seiko technology, inc., all kept in one specific pathogen free facility. For CIA induction, 100. Mu.L of emulsion containing chicken type II collagen (2 mg/ml) and complete Freund's adjuvant (4 mg/ml) was injected intradermally into the tail end. For booster injections, 100 μl of an emulsion containing chicken type II collagen and incomplete freund's adjuvant was injected subcutaneously on day 21. In addition, at day 21, CIA mice of each group were given treatment. Vector group mice were given 100 μl of saline as a control; AZD1208 mice were given AZD1208 (30 mg/kg body weight, dissolved in 100ul saline. Twice weekly for up to day 43.
CIA arthritis scoring criteria: mice were scored every 2 days, recorded, and finally plotted as CIA arthritis scoring curve, starting on day 21 of CIA model induction, with the results shown in fig. 5.
As can be seen from fig. 5: PIM2 kinase small molecule inhibitor AZD1208 was able to ameliorate the arthritic symptoms in CIA arthritic mice, but the effect was not very pronounced.
Example 3
The nanoparticle of the PIM2 kinase small molecule inhibitor wrapped by the neutrophil membrane has the prevention and treatment effect on autoimmune arthritis by inhibiting M1 polarization of BMDM, and the specific process is as follows:
1. in vitro experiments
BMDM cells were plated at 2X 10 per well 5 The individual cells were seeded at a rate in 12-well plates and, after differentiation into mature macrophages by the method described above, were given different stimuli. M0 group was used as control group without any stimulation; LPS (100 ng/mL) was added to M1 group as followsIs a positive control group; the NP group was added with AZD1208 nanoparticles (20 uM); NM group was added to neutrophil membrane (1 mg/mL); the NP@NM-M group was added with a nanoparticle NP@NM-M (20 uM) with a neutrophil membrane encapsulating AZD 1208. MMP2 group added MMP2 polypeptide (20 uM); after 24h stimulation with NP@NM-M (20 uM) and MMP2 (20 uM) added simultaneously to the NP@NM-M+MMP2 group addition, staining was performed for 30min with PE anti-mouse MHC-II antibody and PI anti-mouse F4/80 antibody, and detection was performed using a flow cytometer (thermo cleaner), and the data was analyzed using FlowJo software, as shown in FIG. 6.
As can be seen from fig. 6: NP@NM-M can remarkably inhibit macrophage M1 polarization under the action of MMP2 polypeptide.
2. In vivo test
(1) Prevention and treatment effect of NP@NM-M on autoimmune arthritis
Establishment of CIA arthritis mouse model: DBA mice (8-12 weeks old) were produced by Jiangsu Seiko technology, inc., all kept in one specific pathogen free facility. For CIA induction, 100. Mu.L of emulsion containing chicken type II collagen (2 mg/ml) and complete Freund's adjuvant (4 mg/ml) was injected intradermally into the tail end. For booster injections, 100 μl of an emulsion containing chicken type II collagen and incomplete freund's adjuvant was injected subcutaneously on day 21. In addition, at day 21, CIA mice of each group were given treatment. The Blank group is normal mice that did not induce arthritis; vector group was given 100 μl of physiological saline as a control to the arthritis-induced mice; NM-M group was given pure neutrophil membrane therapy (12 mg/kg body weight, dissolved in 100uL physiological saline) to arthritic mice; NP group was nanoparticle therapy (30 mg/kg body weight, dissolved in 100uL saline) given to AZD1208 of arthritic mice; NP@NM-M is a nanoparticle therapy (30 mg/kg body weight, dissolved in 100uL physiological saline) given to an arthritic mouse with 100uL neutrophil membrane-encapsulated AZD 1208. Oral administration was twice weekly for up to day 43.
CIA arthritis scoring criteria: mice were scored every 2 days, recorded, and finally plotted as CIA arthritis scoring curve, starting on day 21 of CIA model induction, with the results shown in fig. 6.
As can be seen from fig. 7: the NP@NM-M can obviously improve the arthritis symptoms of CIA arthritic mice, and the improvement effect is more obvious than that of free AZD 1208.
(2) Toxic effects of NP@NM-M in vivo
The toxic and side effects of the nanomaterial on mice are judged by taking heart, liver, spleen, lung and kidney tissues after killing each group of CIA arthritis mice in an in-vivo test, embedding the heart, liver, spleen, lung and kidney tissues in paraffin, and then performing HE staining, and the result is shown in figure 8.
As can be seen from fig. 8: NP@NM-M has no obvious toxic or side effect on mice.
The technical scheme of the invention is not limited to the specific embodiment, and all technical modifications made according to the technical scheme of the invention fall within the protection scope of the invention.
Claims (8)
1. A method for preparing a nanoparticle with a neutral granulocyte membrane wrapping PIM2 kinase small molecule inhibitor, which is characterized by comprising the following steps:
s1, obtaining neutrophils and preserving the neutrophils under liquid nitrogen for subsequent membrane separation;
s2, separating neutrophil granulocyte membranes from the neutrophil granulocytes by using a repeated freeze thawing method;
s3, synthesizing nanoparticles of a PIM2 kinase small molecule inhibitor wrapped by a neutrophil membrane:
s31, dissolving 2.5mg of PIM2 kinase small molecule inhibitor AZD1208 in 1ml of dimethyl sulfoxide DMSO to obtain solution A, dissolving 10mg of MMP2 polypeptide in 9ml of water to obtain solution B, mixing the solution A and the solution B, performing ultrasonic treatment on ice in an ultrasonic breaker with the power of 50% for 30min to obtain solution C, dropwise adding the solution C into 10ml of water, and stirring at 4 ℃ for 8-12h to obtain solution D after the dropwise adding is completed within 2 min;
s32, mixing the neutrophil membrane with the solution D according to the volume ratio of the neutrophil membrane to the solution D=1:2.5, performing ultrasonic treatment for 30min, stirring for 2h, extruding liquid by a cell membrane extruder or filtering the liquid by a 0.8 mu m cell filter after extruding by a syringe to obtain the nano-particles of the neutrophil membrane-encapsulated PIM2 kinase small molecular inhibitor.
2. The method for preparing the nanoparticle of the small molecular inhibitor of PIM2 kinase wrapped by the neutrophil membrane according to claim 1, wherein the step S1 specifically comprises:
intraperitoneal injection of 1.5mg/kg lipopolysaccharide LPS in mice to activate neutrophils; after 6h, peripheral blood of the mice is collected in a test tube containing an anticoagulant EDTA-2K, and neutrophils are separated by using a peripheral blood neutrophil separation kit of the mice; the isolated neutrophils were then resuspended in 1M sucrose solution to give neutrophils and stored under liquid nitrogen.
3. The method for preparing the nanoparticle of the small molecular inhibitor of PIM2 kinase encapsulated by the neutrophil membrane according to claim 1, wherein the step S2 specifically comprises:
re-suspending the neutrophils with 1mL pre-chilled double distilled water, then sonicating the mixture at 42kHz for 10min, and performing 3 freeze-thawing cycles; freezing at-80deg.C for 30min, and thawing at room temperature for 30min; centrifuging at 4deg.C under 10000g centrifugal force for 10min, collecting supernatant, and centrifuging at 120000g centrifugal force for 45min; after centrifugation, the neutrophil membrane is obtained and stored at 80 ℃ for standby.
4. The method for preparing the nanoparticle of the small molecular inhibitor of PIM2 kinase wrapped by the neutral granulocyte membrane according to claim 1, wherein the method comprises the following steps: after the solution A and the solution B are mixed in the step S31, 20mg of DSPE-PEG-COOH is added.
5. A nanoparticle of a small molecule inhibitor of PIM2 kinase encapsulated by a neutrophil membrane prepared according to the method of any one of claims 1-4.
6. Use of the nanoparticle of the small molecular inhibitor of PIM2 kinase encapsulated by neutrophil membrane as claimed in claim 5 in the preparation of a medicament for preventing and treating autoimmune arthritis, wherein: the nano-particles for preventing and treating autoimmune arthritis are neutral granulocyte membrane wrapped PIM2 kinase small molecule inhibitor and targeted to inhibit macrophage M1 polarization.
7. The use of the nanoparticle of the small molecular inhibitor of PIM2 kinase wrapped by the medium-neutral granulocyte membrane in the preparation of a medicament for preventing and treating autoimmune arthritis, wherein the nanoparticle is characterized in that: the medicament also comprises pharmaceutically acceptable carriers and/or auxiliary materials.
8. The use of the nanoparticle of the small molecular inhibitor of PIM2 kinase encapsulated by neutrophil membrane in the preparation of a medicament for preventing and treating autoimmune arthritis as claimed in claim 6, wherein: the dosage forms of the medicine comprise injection, oral liquid, tablet, granule, capsule and pill.
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