CN117205154B - Targeted bionic nanometer therapy carrier system for arthritis therapy - Google Patents

Targeted bionic nanometer therapy carrier system for arthritis therapy Download PDF

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CN117205154B
CN117205154B CN202311313669.1A CN202311313669A CN117205154B CN 117205154 B CN117205154 B CN 117205154B CN 202311313669 A CN202311313669 A CN 202311313669A CN 117205154 B CN117205154 B CN 117205154B
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inflammatory
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liposome
phospholipid
carrier system
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CN117205154A (en
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张还添
曾戎
许一迪
彭媛姝
李志鹏
伍昭胜
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First Affiliated Hospital of Jinan University
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/10—Dispersions; Emulsions
    • A61K9/127—Liposomes
    • A61K9/1277—Processes for preparing; Proliposomes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/12—Ketones
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
    • A61K31/19—Carboxylic acids, e.g. valproic acid
    • A61K31/195—Carboxylic acids, e.g. valproic acid having an amino group
    • A61K31/196—Carboxylic acids, e.g. valproic acid having an amino group the amino group being directly attached to a ring, e.g. anthranilic acid, mefenamic acid, diclofenac, chlorambucil
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/337—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having four-membered rings, e.g. taxol
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
    • A61K31/57—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids substituted in position 17 beta by a chain of two carbon atoms, e.g. pregnane or progesterone
    • A61K31/573—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids substituted in position 17 beta by a chain of two carbon atoms, e.g. pregnane or progesterone substituted in position 21, e.g. cortisone, dexamethasone, prednisone or aldosterone
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
    • A61K31/58—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids containing heterocyclic rings, e.g. danazol, stanozolol, pancuronium or digitogenin
    • A61K31/585—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids containing heterocyclic rings, e.g. danazol, stanozolol, pancuronium or digitogenin containing lactone rings, e.g. oxandrolone, bufalin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/24—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing atoms other than carbon, hydrogen, oxygen, halogen, nitrogen or sulfur, e.g. cyclomethicone or phospholipids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/28—Steroids, e.g. cholesterol, bile acids or glycyrrhetinic acid
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

The invention aims to provide a targeted bionic nanometer therapy carrier system which can carry anti-inflammatory substances to more accurately identify and regulate and control macrophages in an inflammatory environment, so as to realize accurate collaborative anti-inflammatory therapy. The provided targeting bionic nanometer therapeutic carrier system is a nanometer liposome composed of phospholipid, active oxygen responsive aryl borate modified phosphatidylserine sodium salt, cholesterol and auxiliary agent; the anti-inflammatory substance is a hydrophobic anti-inflammatory substance or a hydrophilic anti-inflammatory substance. The targeted bionic nano liposome system prepared by the invention is easy to carry anti-inflammatory substances, prevents the mononuclear macrophage system from being cleared in a normal physiological environment, and generates the transformation of simulated apoptosis in an inflammatory environment of an arthritis part, thereby realizing the targeted synergistic anti-inflammatory treatment of macrophages of the arthritis part.

Description

Targeted bionic nanometer therapy carrier system for arthritis therapy
Technical Field
The invention belongs to the field of nanobiotechnology, and particularly relates to a targeted bionic nanotherapeutic carrier system for arthritis treatment.
Background
Inflammatory response is a protective measure of the innate immune system to remove harmful stimuli or pathogens and promote repair, but inflammation inhibition and inflammatory imbalance are closely related to arthritis, atherosclerosis, parkinson's disease, certain tumors, poor regeneration of diseased damaged tissues and the like, and the diseases related to inflammation seriously affect the body health and the quality of life of human beings. Anti-inflammatory is considered an important strategy for controlling these inflammation-related disorders, but how to improve the anti-inflammatory effect, achieving accurate treatment is still a difficulty restricting the success of this strategy.
In recent years, it is recognized that macrophages have extremely strong plasticity and functional diversity, play an important role in pathological processes such as inflammation, and target regulation of polarity and corresponding functions of the macrophages is becoming an important strategy for treating inflammation-related diseases such as arthritis. In view of the fact that macrophages can recognize phosphatidylserine on the surfaces of apoptotic cells to activate phagocytosis and polarization of the apoptotic cells to anti-inflammatory M2 subtype, a synergistic anti-inflammatory therapeutic system targeting macrophages can be constructed by utilizing the biomimetic signal of the apoptotic cells, phosphatidylserine. However, the presence of the mononuclear macrophage system in the normal physiological environment limits the direct targeting of the phosphatidylserine biomimetic therapeutic system to macrophages in the inflammatory environment and limits the therapeutic effect.
Disclosure of Invention
The invention aims to provide a targeted bionic nanometer therapy carrier system which can carry anti-inflammatory substances to more accurately identify and regulate and control macrophages in an inflammatory environment, so as to realize accurate collaborative anti-inflammatory therapy.
The invention firstly provides a targeting bionic nanometer therapeutic carrier system, which is a nanometer liposome composed of phospholipid, active oxygen responsive aryl borate modified phosphatidylserine sodium salt, cholesterol and auxiliary agent;
the phospholipid is one or more of phosphatidylcholine, phosphatidylethanolamine, phosphatidic acid or phosphatidylglycerol;
the active oxygen responsive arylborate modified phosphatidylserine sodium salt is preferably 4-nitrophenyl 4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) benzyl ester modified 1, 2-dipalmitoyl-sn-glycero-3-phospho-L-serine sodium salt (BPS), and has the following structural formula:
the auxiliary agent is one or more of tween, cholic acid, deoxycholate and span; preferably, the auxiliary agent is cholic acid or deoxycholic acid;
in the carrier system, the molar ratio of cholesterol to phospholipid is 1:4-6;
the mol ratio of the auxiliary agent to the phospholipid is 1: 1-40;
the mol ratio of the BPS to the phospholipid is 1:2 to 10;
the bionic nanometer therapeutic carrier system provided by the invention can be used for encapsulating anti-inflammatory substances;
the anti-inflammatory substance is a hydrophobic anti-inflammatory substance or a hydrophilic anti-inflammatory substance;
the anti-inflammatory substance is hydrophobic curcumin, hydrophobic triptolide, hydrophobic paclitaxel, hydrophilic diclofenac sodium, hydrophilic dexamethasone and the like;
the method for preparing BPS provided by the invention comprises the following steps:
(1) Dissolving 1, 2-dipalmitoyl-sn-glycerol-3-phosphate-L-serine sodium salt (PS) in chloroform to obtain a solution A, and dissolving NBC in an organic solvent to obtain a solution B;
(2) Adding organic base into the solution B, reacting for a period of time, adding the solution B into the solution A, stirring for reaction, and regulating the pH value of a reaction medium to be neutral or weak acid;
(3) And (3) purifying and freeze-drying the crude product obtained in the step (2) to obtain the active oxygen responsive BPS pure product.
The preferred scheme of the invention is as follows:
the NBC in the step (1) is 4-nitrophenyl 4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) benzyl ester
The organic solvent in the step (1) is preferably N, N-dimethylformamide, N-dimethylacetamide, dimethyl sulfoxide, tetrahydrofuran or 1,4 dioxane.
The organic base in the step (2) is preferably tetramethyl guanidine, triethylamine, triethylene diamine, N-methylmorpholine, N-diisopropylethylamine, tetramethyl ethylenediamine or ethylenediamine.
The stirring reaction time in the step (2) is preferably 8-24 hours.
The stirring reaction temperature in the step (2) is preferably 20-60 ℃.
The pH regulator in the step (2) is preferably hydrochloric acid or acetic acid.
The purification in the step (3) is preferably dialysis or ultrafiltration.
In a further aspect, the present invention also provides an inflammation therapeutic product prepared by using the targeted bionic nano therapeutic carrier system, where the product is the nano BPS liposome carrier system coated with anti-inflammatory substances;
the preparation method of the therapeutic product, wherein the hydrophobic anti-inflammatory substance is entrapped, comprises the following steps:
1) Dissolving phospholipid, BPS and cholesterol in organic solvent, mixing, adding hydrophobic antiinflammatory material, dissolving again, and mixing;
2) Evaporating the organic solution under vacuum to obtain liposome film;
3) Hydrating the obtained liposome film with physiological saline dissolved with an auxiliary agent to obtain liposome suspension;
4) After the liposome suspension is subjected to ultrasonic treatment, the membrane is coated by an extrusion device to obtain the inflammation treatment product loaded with the hydrophobic drug.
The preparation method of the therapeutic product, wherein the hydrophilic anti-inflammatory substance is entrapped, comprises the following steps:
1) Dissolving phospholipid, BPS and cholesterol in organic solvent, and mixing;
2) Evaporating the organic solution under vacuum to obtain liposome film;
3) Hydrating the obtained liposome film with physiological saline dissolved with an auxiliary agent and a hydrophilic anti-inflammatory substance to obtain liposome suspension;
4) After the liposome suspension is treated by ultrasonic treatment, the membrane is coated by an extrusion device to obtain the inflammation treatment product loaded with the hydrophilic drug.
The organic solvent is one or more of methanol, chloroform or ethanol; preferably chloroform;
the evaporation is preferably performed by a rotary evaporator.
The ultrasonic power is 100W-1000W, and the time is 1-10 min;
the ultrasonic is preferably probe ultrasonic;
the aperture of the extruded membrane is 50 nm-500 nm;
the number of times of extruding the film is 2-6 times.
The targeted bionic nano liposome system prepared by the invention is easy to carry anti-inflammatory substances, prevents the mononuclear macrophage system from being cleared in a normal physiological environment, and generates the transformation of simulated apoptosis in an inflammatory environment of an arthritis part, thereby realizing the targeted synergistic anti-inflammatory treatment of macrophages of the arthritis part.
Drawings
FIG. 1 is an arylboronic acid modification obtained in example 1Molecular formula of 1, 2-dipalmitoyl-sn-glycero-3-phosphate-L-serine sodium salt (BPS) 1 H NMR spectrum.
FIG. 2 is a mass spectrum (negative ion pattern) of arylboronic acid modified 1, 2-dipalmitoyl-sn-glycerol-3-phosphate-L-serine sodium salt (BPS) obtained in example 1.
FIG. 3 is a transmission electron micrograph of the targeted biomimetic nanotherapeutic carrier system obtained in examples 2-4; wherein a is the curcumin-loaded nano-BPS liposome of example 2, b is the dexamethasone-loaded nano-BPS liposome of example 3, and c is the curcumin-loaded nano-BPS liposome of example 4.
FIG. 4 is a graph showing the results of detecting the fraction of PS-signaling-containing liposomes on the surface by a flow cytometer in example 5.
FIG. 5 is a fluorescence photomicrograph of a biomimetic nano-BPS liposome system taken by macrophages in example 6.
FIG. 6 is a graph showing the therapeutic effect of mice model of arthritis of example 7.
Fig. 7 is a graph showing the effect of curcumin-loaded nano-BPS liposomes prepared in example 2 on alleviating OA pain induced by DMM.
Detailed Description
The invention provides a targeting bionic therapeutic nano liposome carrier system which is composed of phospholipid, arylborate modified 1, 2-dipalmitoyl-sn-glycero-3-phosphate-L-serine sodium salt (BPS), cholesterol and an auxiliary agent, and can be loaded with hydrophilic or hydrophobic anti-inflammatory substances.
The present invention will be described in further detail with reference to examples and drawings, but embodiments of the present invention are not limited thereto.
EXAMPLE 1 preparation of arylboronic acid ester modified 1, 2-dipalmitoyl-sn-glycerol-3-phosphate-L-serine sodium salt (BPS)
0.379g (about 0.5 mmol) of 1, 2-dipalmitoyl-sn-glycero-3-phosphate-L-serine sodium salt (PS) was weighed, 15mL of chloroform was added, and an appropriate amount of triethylamine solution was added as solution A; 0.159g (about 0.4 mmol) of NBC,0.035g (about 0.3 mmol) of N-hydroxysuccinimide (NHS) was weighed out and 15mL of DMSO solution was added as solution B.
Solution B was added dropwise to solution a, stirred at room temperature, the reaction monitored by TLC, overnight (time 10 h), after the reaction was completed, the system was alkaline, ph=6.5 was adjusted with 0.5M hydrochloric acid solution, stirred at room temperature for 30 minutes, dialyzed against deionized water for 3 days, and lyophilized to obtain BPS. In CCl 3 D is solvent, which is tested by UX-500 nuclear magnetic resonance 1 The spectrum of H NMR is shown in FIG. 1.
The product BPS was dissolved in CHCl 3 The mass spectrum was tested by using 1290LC-6545QTOF MS high-resolution liquid chromatograph-mass spectrometer, and the negative ion mode spectrogram is shown in figure 2.
Example 2 preparation of curcumin-loaded nano-BPS liposomes
6.5. Mu. Mol of phosphatidylcholine, 1.5. Mu. Mol of BPS, 2. Mu. Mol of cholesterol and 3.75. Mu. Mol of curcumin are dissolved in 5mL of chloroform in sequence and mixed with shaking. Evaporating chloroform by a rotary evaporator to obtain a lipid film. The lipid film was hydrated with 5mL of physiological saline solution containing 0.8. Mu. Mol of sodium deoxycholate to obtain a liposome suspension. After ultrasonic treatment for 5min (300W) with a probe ultrasonic instrument, a lipoeasy LE-800 extruder was used to obtain curcumin-loaded nano BPS liposome suspension by passing through a 100nm polycarbonate membrane three times under argon pressure. The transmission electron micrograph of the obtained curcumin-loaded nano BPS liposome is shown in figure 3a, and the average particle size is about 100 nm.
Example 3 preparation of dexamethasone-loaded nano-BPS liposomes
6.5. Mu. Mol of phosphatidylcholine, 1.5. Mu. Mol of BPS and 2. Mu. Mol of cholesterol were mixed and dissolved in 5mL of chloroform, and the mixture was stirred and mixed uniformly. Evaporating chloroform by a rotary evaporator to obtain a lipid film. The lipid film was hydrated with 5mL of physiological saline solution containing 0.5. Mu. Mol of dexamethasone and 0.8. Mu. Mol of sodium deoxycholate to obtain a liposome suspension. Ultrasonic treatment was performed for 10min (300W) using a probe sonicator, and then a lipobioeasy LE-800 extruder was used to pass through a 100nm polycarbonate membrane six times under nitrogen pressure to obtain dexamethasone-loaded nano BPS liposome suspension. The transmission electron micrograph of the obtained dexamethasone-loaded nano BPS liposome is shown in figure 3b, and the average particle size is about 140 nm.
EXAMPLE 4 curcumin-loaded nano BPS liposome preparation
6.5mmol of phosphatidylcholine, 1.5 mu mol of BPS, 2 mu mol of cholesterol and 3.75 mu mol of curcumin are dissolved in 5mL of chloroform in sequence, and the mixture is mixed by shaking. Evaporating chloroform by a rotary evaporator to obtain a lipid film. The lipid film was hydrated with 5mL of physiological saline solution containing 0.8. Mu. Mol of sodium deoxycholate to obtain a liposome suspension. Ultrasonic treatment is carried out for 5min (100W) by using a probe ultrasonic instrument, then a lipoeasy LE-800 extruder is adopted, and the curcumin-loaded nano BPS liposome is obtained through a 500nm polycarbonate film for two times under the argon pressure, and a transmission electron microscope photograph of the curcumin-loaded nano BPS liposome is shown in figure 3c, and the average particle size is about 500nm.
Example 5 in vitro biomimetic nanotherapeutic System response transition to simulate inflammatory reactive oxygen species environments
According to the principle of specific binding of Annexin V and PS, an Annexin V apoptosis kit containing fluorescein is used for detecting PS signals on the surface of the nano BPS liposome after responsive transformation.
100. Mu.L of curcumin-loaded nano BPS liposome suspension prepared in example 2 was poured into an EP tube, and 5. Mu.L of 5%H was added 2 O 2 Simulating an inflammatory active oxygen environment, and uniformly mixing and reacting for 1 hour; adding an equal volume of Annexin buffer in an apoptosis kit (Kaiyi organism), and uniformly mixing; then 5. Mu.L of annexin V-FITC was added, and after mixing, incubated at room temperature for 30min in the dark. After the incubation, 500. Mu.L of PBS was added for washing, and the mixture was concentrated using an ultrafiltration centrifuge tube having a molecular weight cut-off of 100kDa to remove excess Annexin V-FITC solution. 100. Mu.L of sample in the centrifuge tube was collected and mixed with 200. Mu.L of PBS solution, and the percentage of surface-bound Annexin V-FITC liposomes was measured using a flow cytometer (BD FACSCanto).
Also, respectively without H 2 O 2 The curcumin-loaded nano-BPS liposome prepared in example 2 was treated, and the curcumin-loaded nano-PS liposome prepared by substituting PS for BPS was used as a control group according to the scheme of example 2, and was detected by the above method. As shown in FIG. 4, the results indicate that example 2 was preparedThe drug-loaded nano BPS liposome system is subjected to responsive transformation in an active oxygen environment, and the surface of the drug-loaded nano BPS liposome system is exposed with PS signals.
Example 6 uptake behavior of macrophages on biomimetic nano-BPS liposome systems
First, DHPE/BPS liposomes were prepared as test group 1 using fluorescent marker DHPE instead of curcumin, as per the protocol of example 2, with 5% H 2 O 2 The sample obtained after 1h treatment was used as test group 2, while DHPE/PS liposomes prepared with PS instead of BPS were used as test group 3 (positive control); then, the uptake behavior of macrophages into the above test group was observed using a fluorescence microscope.
The method comprises the following steps: in copolymer Jiao Min, the plank 10 5 Raw264.7 macrophages per well, placed CO 2 The incubator was left overnight. After medium exchange, each group of samples with a total lipid concentration of 500 μm was added, and incubated with macrophages for 1h, all the medium was aspirated, and washed three times with PBS under light-protected conditions. For cell membrane staining, 10. Mu.M of membrane stain DIL was added and the mixture was allowed to act at room temperature for 20min, and washed three times with PBS. And then photographed using a fluorescence microscope. As shown in FIG. 5, raw264.7 macrophages have little uptake of non-H-depleted macrophages 2 O 2 Treated nano BPS liposome system, and through H 2 O 2 The treated nano BPS liposome can be ingested by macrophages due to the exposure of PS, which is consistent with the phenomenon shown by a positive control group.
Example 7 treatment of arthritis model mice
The mouse OA model was successfully established by DMM method, and curcumin-loaded nano BPS liposome prepared in example 2 was injected into tail vein 1 time a week, followed by 5 consecutive times. After the treatment period is finished, the knee joint is decalcified and paraffin is cut into slices, and the damage condition of the joint cartilage of the mice is evaluated by adopting HE and S/O staining, as shown in figure 6, the result shows that the loss of the joint cartilage aminosugar is obviously reduced, and the joint degeneration is delayed.
Meanwhile, as shown in fig. 7, it was observed that the curcumin-loaded nano-BPS liposome prepared in example 2 was injected into tail vein to relieve the pain of OA induced by DMM, and significantly improve animal behavior.
The above examples are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above examples, and any other changes, modifications, substitutions, combinations, and simplifications that do not depart from the spirit and principle of the present invention should be made in the equivalent manner, and the embodiments are included in the protection scope of the present invention.

Claims (4)

1. A bionic nanometer therapeutic carrier system for targeting arthritis is characterized in that the carrier system is a nanometer liposome composed of phospholipid, active oxygen responsive aryl borate modified phosphatidylserine sodium salt (BPS), cholesterol and auxiliary agent,
wherein the structural formula of BPS is as follows:
the aryl borate modified phosphatidylserine sodium salt (BPS) is obtained by reacting 4-nitrophenyl 4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) benzyl ester and 1, 2-dipalmitoyl-sn-glycero-3-phosphoric acid-L-serine sodium salt;
the phospholipid is one or more of phosphatidylcholine, phosphatidylethanolamine, phosphatidic acid or phosphatidylglycerol;
the auxiliary agent is one or more of tween, cholic acid, deoxycholate or span;
the molar ratio of cholesterol to phospholipid in the carrier system is 1:4-6, and the molar ratio of the auxiliary agent to phospholipid is 1: 1-40 parts; the mol ratio of the BPS to the phospholipid is 1: 2-10.
2. An article of manufacture for the treatment of arthritis, wherein the article of manufacture uses the carrier system of claim 1 to carry an anti-inflammatory substance.
3. The article of claim 2, wherein the anti-inflammatory substance is a hydrophobic anti-inflammatory substance, and wherein the article is prepared by a process comprising:
1) Dissolving phospholipid, BPS and cholesterol in organic solvent, mixing, adding hydrophobic antiinflammatory material, dissolving again, and mixing;
2) Evaporating the organic solution under vacuum to obtain liposome film;
3) Hydrating the obtained liposome film with physiological saline dissolved with an auxiliary agent to obtain liposome suspension;
4) After the liposome suspension is subjected to ultrasonic treatment, the membrane is coated by an extrusion device to obtain the inflammation treatment product loaded with the hydrophobic drug.
4. The article of claim 2, wherein the anti-inflammatory substance is a hydrophilic anti-inflammatory substance, and wherein the article is prepared by a process comprising:
1) Dissolving phospholipid, BPS and cholesterol in organic solvent, and mixing;
2) Evaporating the organic solution under vacuum to obtain liposome film;
3) Hydrating the obtained liposome film with physiological saline dissolved with an auxiliary agent and a hydrophilic anti-inflammatory substance to obtain liposome suspension;
4) After the liposome suspension is treated by ultrasonic treatment, the membrane is coated by an extrusion device to obtain the inflammation treatment product loaded with the hydrophilic drug.
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Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105999290A (en) * 2016-04-21 2016-10-12 中国人民解放军第四军医大学 Phosphatidylserine-modified curcumin nanoparticles
CN107082828A (en) * 2017-05-19 2017-08-22 暨南大学 A kind of active oxygen response macromolecule carrier and preparation method thereof
CN111793147A (en) * 2020-08-25 2020-10-20 首都医科大学附属北京中医医院 Modified chitosan, double-response nano-carrier drug, and preparation method and application thereof
CN113081965A (en) * 2021-04-16 2021-07-09 烟台大学 Sensitivity and H based on ROS2S-responsive multifunctional liposome and preparation method and application thereof

Family Cites Families (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR101223366B1 (en) * 2004-05-03 2013-01-16 헤르메스 바이오사이언스, 인코포레이티드 Liposomes useful for drug delivery
US20110020242A1 (en) * 2007-12-12 2011-01-27 Gang Zheng High-density lipoprotein-like peptide-phospholipid scaffold ("hpps") nanoparticles
CN102793668A (en) * 2012-08-30 2012-11-28 绍兴文理学院 PS (Phosphatidylserine)-liposome
CN106074390A (en) * 2016-07-22 2016-11-09 泰山医学院 A kind of Thalidomide liposome nanometer formulation and its preparation method and application
CN112138166B (en) * 2020-09-27 2022-11-25 中国人民解放军军事科学院军事医学研究院 Nano medicine, its preparation method and application
CN115364218A (en) * 2021-05-21 2022-11-22 杭州高田生物医药有限公司 Application of pharmaceutical composition with specific drug-to-lipid ratio in antitumor

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105999290A (en) * 2016-04-21 2016-10-12 中国人民解放军第四军医大学 Phosphatidylserine-modified curcumin nanoparticles
CN107082828A (en) * 2017-05-19 2017-08-22 暨南大学 A kind of active oxygen response macromolecule carrier and preparation method thereof
CN111793147A (en) * 2020-08-25 2020-10-20 首都医科大学附属北京中医医院 Modified chitosan, double-response nano-carrier drug, and preparation method and application thereof
CN113081965A (en) * 2021-04-16 2021-07-09 烟台大学 Sensitivity and H based on ROS2S-responsive multifunctional liposome and preparation method and application thereof

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
含苯硼酸材料在药物投递中的研究进展;吕娟;马如江;史林启;;科学通报;20160710(19);2113-2123 *
纳米氧载体的设计及其在肿瘤诊疗中的应用;于朋;野庆松;吴锦慧;;药学进展;20191125(11);841-855 *

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