CN112220932B - VCAM-1 monoclonal antibody tanshinone IIA nanostructured lipid carrier, preparation method and application - Google Patents
VCAM-1 monoclonal antibody tanshinone IIA nanostructured lipid carrier, preparation method and application Download PDFInfo
- Publication number
- CN112220932B CN112220932B CN202011131715.2A CN202011131715A CN112220932B CN 112220932 B CN112220932 B CN 112220932B CN 202011131715 A CN202011131715 A CN 202011131715A CN 112220932 B CN112220932 B CN 112220932B
- Authority
- CN
- China
- Prior art keywords
- vcam
- tanshinone iia
- monoclonal antibody
- polyethylene glycol
- lipid
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- 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
- A61K47/50—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/68—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
- A61K47/6801—Drug-antibody or immunoglobulin conjugates defined by the pharmacologically or therapeutically active agent
-
- 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
-
- 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
- A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
- A61K47/14—Esters of carboxylic acids, e.g. fatty acid monoglycerides, medium-chain triglycerides, parabens or PEG fatty acid esters
-
- 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
- A61K47/16—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing nitrogen, e.g. nitro-, nitroso-, azo-compounds, nitriles, cyanates
- A61K47/18—Amines; Amides; Ureas; Quaternary ammonium compounds; Amino acids; Oligopeptides having up to five amino acids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P13/00—Drugs for disorders of the urinary system
- A61P13/12—Drugs for disorders of the urinary system of the kidneys
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
- A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Engineering & Computer Science (AREA)
- Medicinal Chemistry (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Epidemiology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Chemical & Material Sciences (AREA)
- Diabetes (AREA)
- Organic Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Hematology (AREA)
- Obesity (AREA)
- Endocrinology (AREA)
- Emergency Medicine (AREA)
- Immunology (AREA)
- Urology & Nephrology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Peptides Or Proteins (AREA)
- Medicinal Preparation (AREA)
Abstract
The invention discloses a VCAM-1 monoclonal antibody tanshinone IIA nanostructured lipid carrier, a preparation method and application thereof. The VCAM-1 monoclonal antibody tanshinone IIA lipid nanoparticle is prepared from 10 parts by mass of tanshinone IIA, 84-89 parts by mass of glyceryl monostearate, 0-5 parts by mass of polyethylene glycol monostearate, 0.5 parts by mass of monoamino terminal polyethylene glycol stearate and 0.5 parts by mass of VCAM-1 monoclonal antibody.
Description
Technical Field
The invention belongs to the field of research on novel drug delivery systems, and particularly relates to a VCAM-1 monoclonal antibody tanshinone IIA lipid nanoparticle, and a preparation method and application thereof.
Background
Diabetic Nephropathy (DN) refers to abnormality of structural, functional or clinical indices of the kidney caused by diabetes, is one of the most common complications of diabetes, and is also the main cause of end-stage renal failure (ESRD). Recent studies have shown that 5.5 billion diabetic patients are expected to occur worldwide by 2030, with 30% to 50% of diabetic patients developing further DN. DN is characterized pathologically by glomerular hypertrophy, thickening of basement membrane and matrix deposition in the mesangial region, with glomerulosclerosis and interstitial fibrosis at the late stages, and even into the uremic phase. To date, clinical treatments for DN have been mainly intervention with drugs that control blood glucose and blood pressure, low protein diet, lipid lowering, and angiotensin, but the overall efficacy has not been satisfactory. Therefore, it is becoming more and more interesting to explore a new therapeutic approach starting from the pathogenesis of DN.
In recent years, it has been found that glomerular endothelial cells are the main target cells for the generation and development of DN. The glomerular endothelial cells are a layer of highly differentiated monolayer flattened cells positioned on the inner wall of blood vessels and lymphatic vessels, and vascular endothelium formed by the endothelial cells is positioned between blood circulation in the lumens and blood vessel walls, is a main barrier for maintaining vascular permeability and is also a target point for regulating glomerular microcirculation by metabolites and hemodynamic signals. When DN occurs, a plurality of factors mainly comprising sugar metabolism disorder can cause endothelial cells to generate a large amount of Reactive Oxygen Radicals (ROS), and the ROS can act on a glomerular basement membrane to ensure that lipid in the basement membrane is overoxidized, so that the permeability of the glomerulus is increased, and protein is easy to deposit on the basement membrane through the endothelial cells; ROS directly acts on endothelial cells, improves the activity of NF-kB, reduces the synthesis of heparan sulfate proteoglycan of glomerular endothelium, thickens a basement membrane or directly destroys glycoprotein of the endothelial cells, thereby damaging the structure and the function of the endothelial cells; ROS can also cause glycosylation or oxidation of antioxidase, so that the activities of antioxidase such as superoxide dismutase, catalase and Gu Zanggan peptide peroxidase are reduced, the capability of an organism in removing free radicals is weakened, and further DN is promoted to be generated and developed. Therefore, starting from reversing the oxidative stress injury of the glomerular endothelial cells, finding a means for effectively treating DN has very important research value and clinical significance.
In view of the great harm of DN to human health, the search for effective DN prevention medicines is the current research focus, the traditional Chinese medicinal materials are explored, the obvious advantages of effective DN prevention and treatment medicines are developed from Chinese herbal medicines, and the Chinese herbal medicines and extracts thereof are widely used for treating various diseases including diabetic nephropathy.
Tanshinone IIA is a fat-soluble effective monomer of Salvia miltiorrhiza Bunge, and has pharmacological effects of resisting oxidation, scavenging oxygen free radicals, inhibiting platelet aggregation and anticoagulation, inhibiting proliferation of smooth muscle cells, dilating coronary artery blood vessel, etc. In recent years, research shows that tanshinone IIA can inhibit the activity of endothelial cell NF-kB induced by hydrogen peroxide, up-regulate the expression of eNOS and other ways to improve the endothelium relaxation dysfunction caused by hyperglycemia; by adjusting the membrane potential of mitochondria, the release of cytochrome C from mitochondria into cytoplasm is inhibited, and the effect of resisting apoptosis is achieved. However, there are still many disadvantages to this drug for DN: the drug has no specific selective action on kidney tissues, so that the distribution of the drug at a target site is low, and the effective treatment of diseases is influenced; accumulation of the drug at non-target sites may increase the incidence of adverse effects of the drug. Therefore, a preparation method is necessary to obviously improve the treatment effect of the tanshinone IIA on DN and reduce the adverse reaction of the tanshinone IIA on the DN from the aspects of increasing the in vivo circulation time of the medicament, changing the in vivo distribution and the like.
The nano drug delivery system has unique advantages in the aspects of realizing targeted distribution of drugs, prolonging the action time of the drugs in vivo and the like: (1) the slow release and the controlled release of the medicine are realized; (2) the targeted delivery of the drug is realized; (3) stable delivery of unstable drugs is achieved; (4) high drug loading of lipophilic drugs is achieved. Based on the characteristics of the nano drug delivery system, the tanshinone IIA can be delivered to the glomerular endothelial cells in a targeted manner by efficiently encapsulating the tanshinone IIA and properly modifying the surface structure of the nano carrier, so that the idea of DN active targeted therapy becomes reality.
DN can induce endothelial cells to produce various biomarkers, such as vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1), vascular Endothelial Growth Factor (VEGF), and the like. VCAM-1 is exposed on the surface of endothelial cells, and the expression level of the VCAM-1 is closely related to the degree of oxidative stress injury, so that the VCAM-1 is applied to the targeted design of nano-carriers by foreign scholars, and the research results of the scholars prove that: the VCAM-1 monoclonal antibody (monoclonal antibody) is modified on the surface of the nano-carrier, and the targeted delivery of the nano-carrier to endothelial cells can be realized by utilizing the principle that the VCAM-1 monoclonal antibody is specifically combined with a VCAM-1 receptor on the surface of the endothelial cells. Kowalski and the like research the influence of whether VCAM-1 monoclonal antibody on the surface of the liposome is modified on the targeted distribution of the VCAM-1 monoclonal antibody, and the result shows that the VCAM-1 monoclonal antibody modified on the surface of the liposome can obviously increase the distribution of the liposome in inflammatory endothelial cells and simultaneously reduce the uptake degree of liver and spleen to the liposome; leus and Gosk et al also demonstrated the specificity of VCAM-1 mAb mediated nanocarriers to specifically target endothelial cells.
Disclosure of Invention
The invention aims to solve the problems in the prior art and provides a VCAM-1 monoclonal antibody tanshinone IIA lipid nanoparticle, a preparation method and an application thereof.
The invention has the following inventive concept: tanshinone IIA is used as a research medicament, a lipid material and polyethylene glycol monostearate are used as main materials, an aqueous solvent diffusion method is used for preparing tanshinone IIA lipid nanoparticles, VCAM-1 monoclonal antibody modification is carried out on the surface of the tanshinone IIA lipid nanoparticles, VCAM-1 monoclonal antibody tanshinone IIA lipid nanoparticles specifically targeting inflammatory vascular endothelial cells are constructed, the improvement effect of the tanshinone IIA lipid nanoparticles on renal function is evaluated at an animal level, and a safe and efficient novel diabetic nephropathy targeted treatment way is searched from the pathogenesis of diabetic nephropathy.
The technical scheme adopted by the invention is as follows:
in a first aspect, the invention provides a tanshinone IIA nanostructured lipid carrier with VCAM-1 monoclonal antibodies, which comprises tanshinone IIA lipid nanoparticles and VCAM-1 monoclonal antibodies;
the tanshinone IIA lipid nanoparticle is prepared from 10 parts by mass of tanshinone IIA, 84-89 parts by mass of glyceryl monostearate, 0-5 parts by mass of polyethylene glycol monostearate and 0.5 part by mass of monoamino-terminated polyethylene glycol stearate;
the amino group of the polyethylene glycol stearate of the single amino terminal on the tanshinone IIA lipid nanoparticle is connected with the carboxyl group on the VCAM-1 monoclonal antibody through N, N' -disuccinimidyl carbonate, so that the antibody is grafted on the lipid nanoparticle.
Preferably, the content of the VCAM-1 monoclonal antibody is 0.5 part by mass, and the mass ratio of the tanshinone IIA lipid nanoparticles to the VCAM-1 monoclonal antibody is 99.5:0.5.
in a second aspect, the present invention provides a method for preparing a nano-structured lipid carrier of VCAM-1 monoclonal antibody tanshinone IIA as described in the first aspect, which comprises the following steps:
1) Mixing 10mg of tanshinone IIA, 84-89mg of glyceryl monostearate, 0-5mg of polyethylene glycol monostearate and 0.5mg of monoamino-terminated polyethylene glycol stearate to obtain a material A;
2) Adding 0.5mL of ethanol into the material A, and heating in a water bath at 60 ℃ to completely melt the material A to obtain a material B;
3) Injecting all the materials B into 9.5mL of deionized water heated in a water bath at 60 ℃ at a stirring speed of 400rpm, stirring for 30s after injection, taking out, and naturally cooling to room temperature to obtain a tanshinone IIA lipid nanoparticle dispersion solution;
4) Adding N, N' -disuccinimidyl carbonate serving as a catalyst into the tanshinone IIA lipid nanoparticle dispersion liquid prepared in the step 3), swirling for 1min to disperse uniformly, and then incubating at room temperature for 3h; then adding 100 mu L of 5mg/mL VCAM-1 monoclonal antibody, swirling for 1min to disperse uniformly, and continuing incubation reaction for 3h at room temperature; and dialyzing and freeze-drying to obtain the VCAM-1 monoclonal antibody tanshinone IIA nanostructured lipid carrier.
Preferably, the polyethylene glycol monostearate has a molecular weight of 2000.
Preferably, the preparation method of the monoamino-terminated polyethylene glycol stearate comprises the following steps: dissolving 59.9mg of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride, 12.0mg of N-hydroxysuccinimide and 17.7mg of stearic acid in 3mL of anhydrous dimethyl sulfoxide, reacting at 400rpm at 60 ℃ for 30min, slowly dropwise adding the reaction solution into a mixed solution prepared from 120mg of amino-terminated polyethylene glycol and 2mL of anhydrous dimethyl sulfoxide, and stirring at 400rpm at room temperature for reacting for 24h; after the reaction is finished, the reaction solution is subjected to dialysis, freeze drying and column chromatography separation and purification to obtain the mono-amino terminal polyethylene glycol stearate.
In a third aspect, the invention provides a nano-structured lipid carrier of VCAM-1 monoclonal antibody tanshinone IIA, prepared by the method of the second aspect.
In a fourth aspect, the invention provides a use of the nano-structured lipid carrier of VCAM-1 monoclonal antibody tanshinone IIA as described in the first and third aspects in the preparation of a targeted drug for diabetic nephropathy.
Compared with the prior art, the invention has the following beneficial effects:
the VCAM-1 monoclonal antibody tanshinone IIA nanostructured lipid carrier provided by the invention can target diabetic nephropathy inflammatory vascular endothelial cells and improve the drug concentration of target cells at the kidney part, thereby improving the drug treatment efficiency, effectively relieving the kidney inflammation level, simultaneously reducing the distribution of drugs at non-target parts and possibly caused adverse reactions, and realizing safe and efficient targeted therapy of diabetic nephropathy.
Drawings
FIG. 1 is a transmission electron microscope photograph of VCAM-1 monoclonal antibody tanshinone IIA lipid nanoparticles;
FIG. 2 is the in vitro release curve of VCAM-1 monoclonal antibody tanshinone IIA lipid nanoparticle;
FIG. 3 is a photograph of the distribution of internal organs of VCAM-1 monoclonal antibody tanshinone IIA lipid nanoparticle.
Detailed Description
The invention will be further elucidated and described with reference to the embodiments and the drawings.
On one hand, the invention provides a VCAM-1 monoclonal antibody tanshinone IIA nanostructured lipid carrier (hereinafter also referred to as VCAM-1 monoclonal antibody tanshinone IIA lipid nanoparticle), which comprises tanshinone IIA lipid nanoparticles and VCAM-1 monoclonal antibody. The tanshinone IIA lipid nanoparticle is prepared from 10 parts by mass of tanshinone IIA, 84-89 parts by mass of glyceryl monostearate, 0-5 parts by mass of polyethylene glycol monostearate (molecular weight of 2000) and 0.5 part by mass of monoamino-terminated polyethylene glycol stearate. The VCAM-1 monoclonal antibody tanshinone IIA lipid nanoparticle is prepared by adding 0.5 part by mass of VCAM-1 monoclonal antibody into the tanshinone IIA lipid nanoparticle, and the amino group of the mono-amino terminal polyethylene glycol stearate on the lipid nanoparticle is connected with the carboxyl group on the VCAM-1 monoclonal antibody through N, N' -disuccinimidyl carbonate, so that the antibody is grafted on the lipid nanoparticle.
On the other hand, the invention provides a preparation method of the VCAM-1 monoclonal antibody tanshinone IIA lipid nanoparticles, which comprises the following steps:
1) Accurately weighing 10% of tanshinone IIA, 84% -89% of glyceryl monostearate and 0% -5% of polyethylene glycol monostearate (molecular weight: 2000 99.5mg of mono-amino terminated polyethylene glycol stearate and 0.5% of the above materials, were mixed to obtain material A;
2) Adding 0.5mL of ethanol into the material A, and heating in a water bath at 60 ℃ to completely melt the material A to obtain a material B;
3) Injecting 0.5mL of material B into 9.5mL of deionized water heated in a water bath at 60 ℃ at a stirring speed of 400rpm by using deionized water as a dispersion medium, stirring for 30s after injection, taking out, and naturally cooling to room temperature to obtain tanshinone IIA liposome nano dispersion;
4) Adding N, N' -disuccinimidyl carbonate into the tanshinone IIA liposome nanoparticle dispersion liquid prepared in the step 3), swirling for 1min to disperse uniformly, and then incubating and reacting at room temperature for 3h; then adding 100 mu L of 5mg/mL VCAM-1 monoclonal antibody, swirling for 1min to disperse uniformly, and continuing incubation reaction for 3h at room temperature; and dialyzing and freeze-drying to obtain the VCAM-1 monoclonal antibody tanshinone IIA lipid nanoparticles. In the reaction, amino groups of polyethylene glycol stearate with single amino terminal on the lipid nanoparticles are connected with carboxyl groups on the VCAM-1 monoclonal antibody through N, N' -disuccinimidyl carbonate, so that the antibody is grafted on the lipid nanoparticles, and the VCAM-1 monoclonal antibody tanshinone IIA lipid nanoparticles are obtained by freeze drying.
The VCAM-1 monoclonal antibody tanshinone IIA lipid nanoparticles prepared by the method are administrated through a mouse tail vein, and the drug has a kidney targeting characteristic in a mouse body, and has a considerable improvement effect on diabetic nephropathy compared with a free tanshinone IIA treatment group and a tanshinone IIA lipid nanoparticle treatment group without VCAM-1 monoclonal antibody modification.
The present invention is further illustrated by the following examples, in which the reagents used may be commercially available products unless otherwise specified. In the following examples, the VCAM-1 monoclonal antibody used was Anti-VCAM1 antibody [ EPR5047] (ab 134047),
in the following examples, the mono-amino terminated polyethylene glycol stearate was prepared by the method: 59.9mg of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride, 12.0mg of N-hydroxysuccinimide and 17.7mg of stearic acid are dissolved in 3mL of anhydrous dimethyl sulfoxide, the mixture reacts for 30min at the temperature of 400rpm and 60 ℃, the reaction solution is slowly dripped into a mixed solution prepared from 120mg of amino-terminated polyethylene glycol and 2mL of anhydrous dimethyl sulfoxide, and the mixture is stirred at the room temperature of 400rpm and reacts for 24h. After the reaction is finished, transferring the reaction solution into a dialysis bag (MWCO: 7 kDa), dialyzing with purified water for 48h, removing water-soluble byproducts, freeze-drying to obtain a coarse product of the polyethylene glycol stearate with the single amino terminal, and purifying the obtained freeze-dried product by adopting a column chromatography separation method to obtain the polyethylene glycol stearate with the single amino terminal. Of course, this is only an alternative method of preparation and other examples may employ the commercially available finished mono amino-terminated polyethylene glycol stearate.
The first embodiment is as follows: preparing a VCAM-1 monoclonal antibody tanshinone IIA nanostructured lipid carrier: accurately weighing 99.5mg of tanshinone IIA, 84mg of glyceryl monostearate, 5mg of polyethylene glycol monostearate and 0.5mg of monoamino-terminated polyethylene glycol stearate, adding 0.5mL of ethanol to dissolve the materials to obtain an organic phase, heating the organic phase in a water bath at 60 ℃ to completely melt the materials, taking deionized water as a dispersion medium, rapidly injecting the organic phase into 9.5mL of deionized water heated in the water bath at 60 ℃ in turn at a stirring speed of 400rpm, taking out the materials after stirring for 30s after the injection is finished, and naturally cooling the materials to room temperature to obtain the tanshinone IIA lipid nanoparticles. Adding 0.1mg of N, N' -disuccinimidyl carbonate as a catalyst, performing vortex for 1min to disperse uniformly, performing incubation reaction at room temperature for 3h, adding 100 mu L of VCAM-1 monoclonal antibody (5 mg/mL), performing vortex for 1min to disperse uniformly, and performing incubation reaction at room temperature for 3h. The reaction comprises the steps of connecting amino of polyethylene glycol on a drug carrier and carboxyl on a VCAM-1 monoclonal antibody through N, N' -disuccinimidyl carbonate, successfully grafting the antibody onto the drug carrier, and dialyzing and freeze-drying to obtain the VCAM-1 monoclonal antibody tanshinone IIA lipid nanoparticles.
The particle size of the prepared nanoparticles is measured by a dynamic light scattering method. Diluting the sample to 0.1mg/mL with deionized water, and determining the particle size of VCAM-1 monoclonal antibody tanshinone IIA lipid nanoparticles to be 43.8 +/-9.5 nm by using a Malvern Zetasizer Nano-S90 nanometer particle size analyzer.
The encapsulation efficiency and the drug-loading rate of the VCAM-1 monoclonal antibody tanshinone IIA lipid nanoparticles are measured by an acid-adjusting centrifugal method and a high-performance liquid phase method. Adding a proper amount of 0.1M hydrochloric acid into the VCAM-1 monoclonal antibody tanshinone IIA lipid nanoparticles which are placed to room temperature after the preparation is finished, reducing the pH of the system to 1.2 to promote the flocculation of the nanoparticles, centrifuging at 20000rpm for 30min, measuring the drug concentration in the supernatant by HPLC, and calculating the encapsulation rate and the drug loading rate of the VCAM-1 monoclonal antibody tanshinone IIA lipid nanoparticles according to the following formula.
Encapsulation efficiency = (drug mass in lipid nanoparticle/total drug dosage) × 100%
Drug loading = (drug mass in lipid nanoparticles/total mass of drug-loaded nanoparticles) × 100%
Through determination, the encapsulation rate and the drug-loading rate of the VCAM-1 monoclonal antibody tanshinone IIA lipid nanoparticles are 86.36 percent and 4.39 percent respectively.
Example two: preparing the tanshinone IIA lipid nanoparticles with VCAM-1 monoclonal antibody, precisely weighing 10mg of tanshinone IIA, 85mg of glyceryl monostearate, 4mg of polyethylene glycol monostearate and 0.5mg of monoamino-terminated polyethylene glycol stearate, wherein the total amount is 99.5mg, adding 0.5mL of ethanol to dissolve the components to obtain an organic phase, heating the organic phase in a water bath at 60 ℃ to completely melt the materials, using deionized water as a dispersion medium, rapidly injecting the organic phase into 9.5mL of deionized water heated in the water bath at 60 ℃ in turn at a stirring speed of 400rpm, taking out the mixture after stirring for 30s after the injection is finished, and naturally cooling the mixture to room temperature to obtain the tanshinone IIA lipid nanoparticles. Adding 0.1mg of N, N' -disuccinimidyl carbonate serving as a catalyst, performing vortex for 1min to disperse uniformly, performing incubation reaction at room temperature for 3h, adding 100 mu L of VCAM-1 monoclonal antibody (5 mg/mL), performing vortex for 1min to disperse uniformly, and performing incubation reaction at room temperature for 3h. The reaction comprises the steps of connecting amino of polyethylene glycol on a drug carrier and carboxyl on a VCAM-1 monoclonal antibody through N, N' -disuccinimidyl carbonate, successfully grafting the antibody on the drug carrier, and dialyzing and freeze-drying to obtain the VCAM-1 monoclonal antibody tanshinone IIA lipid nanoparticles.
The particle size of the prepared nanoparticles is measured by a dynamic light scattering method. Diluting the sample to 0.1mg/mL with deionized water, and determining the particle size of VCAM-1 monoclonal antibody tanshinone IIA lipid nanoparticles to be 40.3 +/-10.3 nm by using a Malvern Zetasizer Nano-S90 nanometer particle size analyzer.
The encapsulation efficiency and drug-loading capacity of the tanshinone IIA lipid nanoparticles are measured by an acid-adjusting centrifugation method in combination with a high performance liquid chromatography method. Adding appropriate amount of 0.1M hydrochloric acid into the VCAM-1 monoclonal antibody tanshinone IIA nanoparticles which are placed to room temperature after the preparation is finished, reducing the pH of the system to 1.2 to promote the flocculation of the nanoparticles, centrifuging at 20000rpm for 30min, measuring the drug concentration in the supernatant by HPLC, and calculating the encapsulation rate and the drug loading rate of the tanshinone IIA lipid nanoparticles according to the following formula.
Encapsulation efficiency = (drug mass in lipid nanoparticle/total drug dosage) × 100%
Drug loading = (drug mass in lipid nanoparticles/total mass of drug-loaded nanoparticles) × 100%
Through determination, the encapsulation efficiency and the drug-loading rate of the VCAM-1 monoclonal antibody tanshinone IIA lipid nanoparticles are 84.97% and 4.74% respectively.
Example three: preparing the tanshinone IIA lipid nanoparticles with VCAM-1 monoclonal antibody, precisely weighing 10mg of tanshinone IIA, 86mg of glyceryl monostearate, 3mg of polyethylene glycol monostearate and 0.5mg of monoamino-terminated polyethylene glycol stearate, wherein the total amount is 99.5mg, adding 0.5mL of ethanol to dissolve the components to obtain an organic phase, heating the organic phase in a water bath at 60 ℃ to completely melt the materials, using deionized water as a dispersion medium, rapidly injecting the organic phase into 9.5mL of deionized water heated in the water bath at 60 ℃ at a stirring speed of 400rpm, taking out the mixture after stirring for 30s after the injection is finished, and naturally cooling the mixture to room temperature to obtain the tanshinone IIA lipid nanoparticles. Adding 0.1mg of N, N' -disuccinimidyl carbonate serving as a catalyst, performing vortex for 1min to disperse uniformly, performing incubation reaction at room temperature for 3h, adding 100 mu L of VCAM-1 monoclonal antibody (5 mg/mL), performing vortex for 1min to disperse uniformly, and performing incubation reaction at room temperature for 3h. The reaction comprises the steps of connecting amino of polyethylene glycol on a drug carrier and carboxyl on a VCAM-1 monoclonal antibody through N, N' -disuccinimidyl carbonate, successfully grafting the antibody onto the drug carrier, and dialyzing and freeze-drying to obtain the VCAM-1 monoclonal antibody tanshinone IIA lipid nanoparticles.
The particle size of the prepared nanoparticles is measured by a dynamic light scattering method. Diluting the sample to 0.1mg/mL with deionized water, and measuring the particle sizes of VCAM-1 monoclonal antibody tanshinone IIA lipid nanoparticles to be 38.5 +/-7.8 nm by using a Malvern Zetasizer Nano-S90 nanometer particle size analyzer.
The encapsulation efficiency and the drug loading capacity of the VCAM-1 monoclonal antibody tanshinone IIA lipid nanoparticles are measured by an acid-regulating centrifugation method and a high performance liquid chromatography method. Adding a proper amount of 0.1M hydrochloric acid into the VCAM-1 monoclonal antibody tanshinone IIA nanoparticles which are placed to room temperature after the preparation is finished, reducing the pH of the system to 1.2 to promote the flocculation of the nanoparticles, centrifuging at 20000rpm for 30min, measuring the drug concentration in the supernatant by HPLC, and calculating the encapsulation rate and the drug loading rate of the VCAM-1 monoclonal antibody tanshinone IIA lipid nanoparticles according to the following formula.
Encapsulation efficiency = (drug mass in lipid nanoparticle/total drug dosage) × 100%
Drug loading rate = (drug mass in lipid nanoparticle/total mass of drug-loaded nanoparticle) × 100%
Through determination, the encapsulation efficiency and the drug-loading rate of the VCAM-1 monoclonal antibody tanshinone IIA lipid nanoparticles are 88.36% and 5.03% respectively.
Example four: preparing the tanshinone IIA lipid nanoparticles with VCAM-1 monoclonal antibody, precisely weighing 10mg of tanshinone IIA, 87mg of glyceryl monostearate, 2mg of polyethylene glycol monostearate and 0.5mg of monoamino-terminated polyethylene glycol stearate, wherein the total amount is 99.5mg, adding 0.5mL of ethanol to dissolve the materials to obtain an organic phase, heating the organic phase in a water bath at 60 ℃ to completely melt the materials, using deionized water as a dispersion medium, rapidly injecting the organic phase into 9.5mL of deionized water heated in the water bath at 60 ℃ at a stirring speed of 400rpm, taking out the mixture after stirring for 30s after the injection is finished, and naturally cooling the mixture to room temperature to obtain the tanshinone IIA lipid nanoparticles. Adding 0.1mg of N, N' -disuccinimidyl carbonate as a catalyst, performing vortex for 1min to disperse uniformly, performing incubation reaction at room temperature for 3h, adding 100 mu L of VCAM-1 monoclonal antibody (5 mg/mL), performing vortex for 1min to disperse uniformly, and performing incubation reaction at room temperature for 3h. The reaction comprises the steps of connecting amino of polyethylene glycol on a drug carrier and carboxyl on a VCAM-1 monoclonal antibody through N, N' -disuccinimidyl carbonate, successfully grafting the antibody onto the drug carrier, and dialyzing and freeze-drying to obtain the VCAM-1 monoclonal antibody tanshinone IIA lipid nanoparticles.
The particle size of the prepared nanoparticles is measured by a dynamic light scattering method. Diluting the sample to 0.1mg/mL with deionized water, and measuring the particle sizes of VCAM-1 monoclonal antibody tanshinone IIA lipid nanoparticles to be 36.1 +/-6.8 nm by using a Malvern Zetasizer Nano-S90 nanometer particle size analyzer.
The encapsulation efficiency and the drug-loading rate of the VCAM-1 monoclonal antibody tanshinone IIA lipid nanoparticles are measured by an acid-adjusting centrifugal method and a high-performance liquid phase method. Adding a proper amount of 0.1M hydrochloric acid into the VCAM-1 monoclonal antibody tanshinone IIA nanoparticles which are placed to room temperature after the preparation is finished, reducing the pH of the system to 1.2 to promote the flocculation of the nanoparticles, centrifuging at 20000rpm for 30min, measuring the drug concentration in the supernatant by HPLC, and calculating the encapsulation rate and the drug loading rate of the VCAM-1 monoclonal antibody tanshinone IIA lipid nanoparticles according to the following formula.
Encapsulation efficiency = (drug mass in lipid nanoparticle/total drug dosage) × 100%
Drug loading rate = (drug mass in lipid nanoparticle/total mass of drug-loaded nanoparticle) × 100%
Through determination, the encapsulation efficiency and the drug-loading rate of the VCAM-1 monoclonal antibody tanshinone IIA lipid nanoparticles are 86.24% and 4.25% respectively.
Example five: preparing the tanshinone IIA lipid nanoparticles with VCAM-1 monoclonal antibody, precisely weighing 10mg of tanshinone IIA, 88mg of glyceryl monostearate, 1mg of polyethylene glycol monostearate and 0.5mg of monoamino-terminated polyethylene glycol stearate, wherein the total amount is 99.5mg, adding 0.5mL of ethanol to dissolve the materials to obtain an organic phase, heating the organic phase in a water bath at 60 ℃ to completely melt the materials, using deionized water as a dispersion medium, rapidly injecting the organic phase into 9.5mL of deionized water heated in the water bath at 60 ℃ at a stirring speed of 400rpm, taking out the mixture after stirring for 30s after the injection is finished, and naturally cooling the mixture to room temperature to obtain the tanshinone IIA lipid nanoparticles. Adding 0.1mg of N, N' -disuccinimidyl carbonate as a catalyst, performing vortex for 1min to disperse uniformly, performing incubation reaction at room temperature for 3h, adding 100 mu L of VCAM-1 monoclonal antibody (5 mg/mL), performing vortex for 1min to disperse uniformly, and performing incubation reaction at room temperature for 3h. The reaction comprises the steps of connecting amino of polyethylene glycol on a drug carrier and carboxyl on a VCAM-1 monoclonal antibody through N, N' -disuccinimidyl carbonate, successfully grafting the antibody onto the drug carrier, and dialyzing and freeze-drying to obtain the VCAM-1 monoclonal antibody tanshinone IIA lipid nanoparticles.
The particle size of the prepared nanoparticles is measured by a dynamic light scattering method. Diluting the sample to 0.1mg/mL with deionized water, and determining the particle sizes of VCAM-1 monoclonal antibody tanshinone IIA lipid nanoparticles to be 33.5 +/-6.2 nm by using a Malvern Zetasizer Nano-S90 nanometer particle size analyzer.
The encapsulation efficiency and the drug loading capacity of the VCAM-1 monoclonal antibody tanshinone IIA lipid nanoparticles are measured by an acid-regulating centrifugation method and a high performance liquid chromatography method. Adding a proper amount of 0.1M hydrochloric acid into the VCAM-1 monoclonal antibody tanshinone IIA nanoparticles which are placed to room temperature after the preparation is finished, reducing the pH of the system to 1.2 to promote the flocculation of the nanoparticles, centrifuging at 20000rpm for 30min, measuring the drug concentration in the supernatant by HPLC, and calculating the encapsulation rate and the drug loading rate of the VCAM-1 monoclonal antibody tanshinone IIA lipid nanoparticles according to the following formula.
Encapsulation efficiency = (drug mass in lipid nanoparticle/total drug dosage) × 100%
Drug loading rate = (drug mass in lipid nanoparticle/total mass of drug-loaded nanoparticle) × 100%
Through determination, the encapsulation rate and the drug-loading rate of the VCAM-1 monoclonal antibody tanshinone IIA lipid nanoparticles are respectively 89.52 percent and 5.54 percent.
Example six: the preparation method of the VCAM-1 monoclonal antibody tanshinone IIA lipid nanoparticle comprises the steps of precisely weighing 10mg of tanshinone IIA, 89mg of glyceryl monostearate and 0.5mg of mono-amino terminal polyethylene glycol stearate, wherein the total amount is 100mg, adding 0.5mL of ethanol to dissolve the mixture to obtain an organic phase, heating the organic phase in a water bath at 60 ℃ to completely melt the material, taking deionized water as a dispersion medium, rapidly injecting the organic phase into 9.5mL of deionized water heated in the water bath at 60 ℃ in turn at a stirring speed of 400rpm, taking out the mixture after stirring for 30s after the injection is finished, and naturally cooling the mixture to room temperature to obtain the tanshinone IIA lipid nanoparticle. Adding 0.1mg of N, N' -disuccinimidyl carbonate serving as a catalyst, performing vortex for 1min to disperse uniformly, performing incubation reaction at room temperature for 3h, adding 100 mu L of VCAM-1 monoclonal antibody (5 mg/mL), performing vortex for 1min to disperse uniformly, and performing incubation reaction at room temperature for 3h. The reaction comprises the steps of connecting amino of polyethylene glycol on a drug carrier and carboxyl on a VCAM-1 monoclonal antibody through N, N' -disuccinimidyl carbonate, successfully grafting the antibody onto the drug carrier, and dialyzing and freeze-drying to obtain the VCAM-1 monoclonal antibody tanshinone IIA lipid nanoparticles.
The particle size of the prepared nanoparticles is measured by a dynamic light scattering method. Diluting the sample to 0.1mg/mL with deionized water, and determining the particle sizes of VCAM-1 monoclonal antibody tanshinone IIA lipid nanoparticles to be 30.1 +/-5.2 nm by using a Malvern Zetasizer Nano-S90 nanometer particle size analyzer.
The encapsulation efficiency and the drug loading capacity of the VCAM-1 monoclonal antibody tanshinone IIA lipid nanoparticles are measured by an acid-regulating centrifugation method and a high performance liquid chromatography method. Adding a proper amount of 0.1M hydrochloric acid into the VCAM-1 monoclonal antibody tanshinone IIA nanoparticles which are placed to room temperature after the preparation is finished, reducing the pH of the system to 1.2 to promote the flocculation of the nanoparticles, centrifuging at 20000rpm for 30min, measuring the drug concentration in the supernatant by HPLC, and calculating the encapsulation rate and the drug loading rate of the VCAM-1 monoclonal antibody tanshinone IIA lipid nanoparticles according to the following formula.
Encapsulation efficiency = (drug mass in lipid nanoparticle/total drug dosage) × 100%
Drug loading rate = (drug mass in lipid nanoparticle/total mass of drug-loaded nanoparticle) × 100%
Through determination, the encapsulation efficiency and the drug-loading rate of the VCAM-1 monoclonal antibody tanshinone IIA lipid nanoparticles are 88.36% and 5.25% respectively.
Example six: and (3) observing the form of the VCAM-1 monoclonal antibody tanshinone IIA lipid nanoparticles. The VCAM-1 monoclonal antibody tanshinone IIA lipid nanoparticles in the first embodiment are taken to prepare a dispersion liquid of 0.1mg/mL, the lipid nanoparticles are dispersed and dripped on a copper mesh, the dispersion liquid is naturally dried, 1% uranyl acetate solution is used for negative staining for 1min, and a JEM-1200EX type transmission electron microscope is used for observing the shape, and the result is shown in figure 1 and is in a similar spherical shape.
Example seven: and (3) carrying out in-vitro drug release investigation on the VCAM-1 monoclonal antibody tanshinone IIA lipid nanoparticles. The VCAM-1 monoclonal antibody tanshinone IIA lipid nanoparticles in example one were used for in vitro drug release investigation, and free tanshinone IIA was used as a control. 1mL of VCAM-1 monoclonal antibody tanshinone IIA lipid nanoparticle solution of free tanshinone IIA and tanshinone IIA with equal mass is respectively taken and placed in a dialysis bag, and then placed in a release tube containing 15mL of PBS with pH value of 7.4, and the solution is vibrated at the constant temperature of 37 ℃ and 60 rpm. 1mL was sampled at the set time point and 1mL of fresh PBS solution was made up. And (3) measuring the concentration of the tanshinone IIA in the release medium by a high performance liquid chromatography method, and calculating the cumulative release amount of the tanshinone IIA. The release behavior of VCAM-1 monoclonal antibody tanshinone IIA lipid nanoparticles is determined as shown in fig. 2, and compared with a simple free drug, the lipid nanoparticles have a sustained release effect on the encapsulated tanshinone IIA.
Example eight: the kidney targeting research of the VCAM-1 monoclonal antibody tanshinone IIA lipid nanoparticles. Taking a c57bl/6 mouse as a model animal, performing left side kidney ligation, recovering for 1 week after operation, feeding for 4 weeks with high fat feed, and performing intraperitoneal injection of l% Streptozotocin (STZ) 25mg/kg to the molded animal (operation and high fat) at one time. After 72 hours, fasting is performed for 3 hours, and the blood sugar of the tail tip of the animal is greater than 16.7mmol/L for the test.
The VCAM-1 monoclonal antibody tanshinone IIA lipid nanoparticles in the first embodiment are marked by a near-infrared fluorescent probe Dir, and the distribution condition of the nanoparticles in a model animal body is researched. The results are shown in fig. 3, which shows that after the nanostructured lipid carrier is modified by the VCAM-1 monoclonal antibody, the kidney targeting distribution is significantly improved, which indicates that the vascular endothelial cells of the kidney tissue of the model animal with diabetic nephropathy highly express the VCAM-1 receptor, the VCAM-1 monoclonal antibody nano-carrier is specifically bound by the antibody receptor and is distributed in a large amount in the kidney of the model mouse, and the VCAM-1 monoclonal antibody nano-carrier is distributed more in the kidney of the model mouse than the lipid nanoparticles without the VCAM-1 monoclonal antibody modification, which shows the kidney targeting property of the VCAM-1 monoclonal antibody tanshinone IIA lipid nanoparticles.
Example nine: pharmacodynamics research of VCAM-1 monoclonal antibody tanshinone IIA lipid nanoparticle on diabetic nephropathy. A left kidney ligation technique is adopted, and streptozotocin is injected into the abdominal cavity and high-fat feed are used for feeding to construct an animal model of diabetic nephropathy. Collecting 24h urine by a metabolism cage, recording urine volume, determining 24h urine volume and 24h urine protein quantification according to the kit operation instructions, and calculating glomerular filtration rate; the levels of serum creatinine and urea nitrogen were determined using an automated biochemical analyzer. The experimental groups were as follows: (1) normal; (2) DN + physiological saline; (3) DN + tanshinone IIA; (4) DN + tanshinone IIA lipid nanoparticles of example one; (5) DN + tanshinone IIA lipid nanoparticles of VCAM-1 monoclonal antibody in example I. The results are shown in table 1 below, which,
TABLE 1 Effect of the various formulations on glomerular filtration Rate, serum creatinine and Urea Nitrogen in mice with diabetic nephropathy
The above-described embodiments are merely preferred embodiments of the present invention, which should not be construed as limiting the invention. Various changes and modifications may be made by one of ordinary skill in the pertinent art without departing from the spirit and scope of the present invention. Therefore, the technical scheme obtained by adopting the mode of equivalent replacement or equivalent transformation is within the protection scope of the invention.
Claims (5)
1. A VCAM-1 monoclonal antibody tanshinone IIA nanostructured lipid carrier is characterized by comprising tanshinone IIA lipid nanoparticles and a VCAM-1 monoclonal antibody;
the tanshinone IIA lipid nanoparticle is prepared from 10 parts by mass of tanshinone IIA, 84-89 parts by mass of glyceryl monostearate, 0-5 parts by mass of polyethylene glycol monostearate and 0.5 part by mass of monoamino-terminated polyethylene glycol stearate;
the amino group of the polyethylene glycol stearate of the single amino terminal on the tanshinone IIA lipid nanoparticle is connected with the carboxyl group on the VCAM-1 monoclonal antibody through N, N' -disuccinimidyl carbonate, so that the antibody is grafted on the lipid nanoparticle;
the preparation method of the VCAM-1 monoclonal antibody tanshinone IIA nanostructured lipid carrier comprises the following steps:
1) Mixing 10mg of tanshinone IIA, 84-89mg of glyceryl monostearate, 0-5mg of polyethylene glycol monostearate and 0.5mg of monoamino-terminated polyethylene glycol stearate to obtain a material A;
2) Adding 0.5mL ethanol into the material A, and heating in a water bath at 60 ℃ to completely melt the material A to obtain a material B;
3) Injecting all the materials B into 9.5mL deionized water heated in a water bath at 60 ℃ at a stirring speed of 400rpm, stirring 30s after injection, taking out, and naturally cooling to room temperature to obtain a tanshinone IIA lipid nanoparticle dispersion liquid;
4) Adding N, N' -disuccinimidyl carbonate serving as a catalyst into the tanshinone IIA lipid nanoparticle dispersion liquid prepared in the step 3), swirling for 1min to disperse uniformly, and then carrying out incubation reaction at room temperature for 3h; then adding 100 mu L of 5mg/mL VCAM-1 monoclonal antibody, swirling for 1min to disperse uniformly, and then continuing incubating the reaction for 3h at room temperature; and dialyzing and freeze-drying to obtain the VCAM-1 monoclonal antibody tanshinone IIA nanostructured lipid carrier.
2. The lipid carrier with a nano-structure of VCAM-1 monoclonal antibody tanshinone IIA as claimed in claim 1, wherein the content of the VCAM-1 monoclonal antibody is 0.5 parts by mass, and the mass ratio of the tanshinone IIA lipid nanoparticle to the VCAM-1 monoclonal antibody is 99.5:0.5.
3. the nano-structured lipid carrier of VCAM-1 mab tanshinone IIA as claimed in claim 1, wherein the molecular weight of polyethylene glycol monostearate is 2000.
4. The lipid carrier with a nano-structure of VCAM-1 monoclonal antibody tanshinone IIA as claimed in claim 1, wherein the preparation method of the mono-amino terminal polyethylene glycol stearate comprises: dissolving 59.9mg of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride, 12.0mg of N-hydroxysuccinimide and 17.7mg stearic acid in 3mL of anhydrous dimethyl sulfoxide, reacting at 400rpm at 60 ℃ for 30min, slowly dropwise adding a reaction solution into a mixed solution prepared from 120mg of double-ended aminopolyethylene glycol and 2mL of anhydrous dimethyl sulfoxide, and reacting at room temperature under 400rpm stirring for 24h; after the reaction is finished, the reaction solution is subjected to dialysis, freeze drying and column chromatography separation and purification to obtain the mono-amino terminal polyethylene glycol stearate.
5. An application of the VCAM-1 monoclonal antibody tanshinone IIA nanostructured lipid carrier of any of claims 1~4 in the preparation of a targeted medicament for diabetic nephropathy.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011131715.2A CN112220932B (en) | 2020-10-21 | 2020-10-21 | VCAM-1 monoclonal antibody tanshinone IIA nanostructured lipid carrier, preparation method and application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011131715.2A CN112220932B (en) | 2020-10-21 | 2020-10-21 | VCAM-1 monoclonal antibody tanshinone IIA nanostructured lipid carrier, preparation method and application |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN112220932A CN112220932A (en) | 2021-01-15 |
| CN112220932B true CN112220932B (en) | 2022-12-13 |
Family
ID=74118289
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202011131715.2A Active CN112220932B (en) | 2020-10-21 | 2020-10-21 | VCAM-1 monoclonal antibody tanshinone IIA nanostructured lipid carrier, preparation method and application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112220932B (en) |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106074389A (en) * | 2016-06-30 | 2016-11-09 | 浙江大学 | ICAM 1 monoclonal antibody modifies simvastatin nano structured lipid carrier and preparation and application |
-
2020
- 2020-10-21 CN CN202011131715.2A patent/CN112220932B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106074389A (en) * | 2016-06-30 | 2016-11-09 | 浙江大学 | ICAM 1 monoclonal antibody modifies simvastatin nano structured lipid carrier and preparation and application |
Non-Patent Citations (2)
| Title |
|---|
| Ganesh Ram R. Visweswaran等.Targeting Rapamycin to Podocytes Using a Vascular Cell Adhesion Molecule-1 (VCAM-1)-Harnessed SAINT-Based Lipid Carrier System.《PLOS ONE》.2015,1-17. * |
| 瞿继兰等.丹参酮ⅡA 固体脂质纳米粒的制备及质量评价.《天津中医药》.2014,564-566. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN112220932A (en) | 2021-01-15 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107952072B (en) | Preparation method of drug-loaded and oxygen-loaded hybrid protein nanoparticles, drug-loaded and oxygen-loaded hybrid protein nanoparticles and application | |
| CN111617246B (en) | Self-assembled nanoparticles of pure photosensitizer and preparation and application thereof | |
| CN112716915A (en) | Bionic nano-carrier and application thereof in preparing medicament for treating brain glioma | |
| WO2023125404A1 (en) | Spinal cord injury targeted drug, polymer-hydrophobic compound micelle, and preparation method for spinal cord injury targeted drug | |
| CN113876716B (en) | Bioadhesive nanoparticle for treating gastrointestinal diseases and preparation method thereof | |
| Shen et al. | Biomimetic nanoparticles: U937 cell membranes based core–shell nanosystems for targeted atherosclerosis therapy | |
| CN107714675B (en) | Gambogic acid core-shell structure composite nano preparation and preparation method thereof | |
| CN111494338B (en) | Ovalbumin nanocapsule with synergistic effect of chemotherapy and photodynamic therapy and preparation method and application thereof | |
| CN112220932B (en) | VCAM-1 monoclonal antibody tanshinone IIA nanostructured lipid carrier, preparation method and application | |
| Hu et al. | Ultrasound-assisted biomimetic nanobubbles for targeted treatment of atherosclerosis | |
| CN104045823B (en) | A kind of Enoxolone derivative and its preparation method and application | |
| Zhu et al. | Construction of long circulating and deep tumor penetrating gambogic acid-hydroxyethyl starch nanoparticles | |
| CN109953974B (en) | Preparation method of enzyme-reduction dual-responsiveness hyaluronic acid-polypropylene sulfide copolymer nanocapsule | |
| CN109662956B (en) | Application of oleanolic acid grafted chitosan drug-loaded nanoparticles | |
| WO2023193389A1 (en) | Resveratrol-lecithin nanoparticle, method for preparing same, and use thereof | |
| CN114209663B (en) | Preparation and application of phloretin-loaded soybean lecithin-chitosan nanoparticles for preventing diabetes | |
| CN110464708A (en) | A kind of spirulina nanometer formulation and preparation method thereof | |
| Liang et al. | The therapeutic potential of targeted Nanoparticulate systems to treat rheumatoid arthritis | |
| CN113577016A (en) | Triptolide-gold nanoparticle/hyaluronic acid composite hydrogel and preparation and application thereof | |
| CN114668856A (en) | Application of beta-cyclodextrin mediated immune cell-drug loaded liposome couplet in resisting atherosclerosis | |
| CN106822037A (en) | A kind of preparation method of the nano-particle of the tripeptides modification for targetting colitis tissue | |
| CN109289057B (en) | Dexamethasone nanometer preparation for targeted therapy of rheumatoid arthritis and preparation method thereof | |
| CN111358955A (en) | Inflammation-targeted bindarit nanoparticle for treating lipid metabolism diseases, and preparation method and application thereof | |
| Zhang et al. | Cyclic RGD functionalized PLGA nanoparticles loaded with noncovalent complex of indocyanine green with urokinase for synergistic thrombolysis | |
| CN115737598B (en) | Preparation of nanoparticle cluster Al-PHNPs-PAMAM and application thereof in treating brain diseases |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |