CN106074389A - ICAM 1 monoclonal antibody modifies simvastatin nano structured lipid carrier and preparation and application - Google Patents

ICAM 1 monoclonal antibody modifies simvastatin nano structured lipid carrier and preparation and application Download PDF

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CN106074389A
CN106074389A CN201610530175.2A CN201610530175A CN106074389A CN 106074389 A CN106074389 A CN 106074389A CN 201610530175 A CN201610530175 A CN 201610530175A CN 106074389 A CN106074389 A CN 106074389A
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simvastatin
icam
monoclonal antibody
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姜赛平
李淑娟
杜永忠
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Zhejiang University ZJU
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    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
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Abstract

The present invention provides a kind of ICAM 1 monoclonal antibody to modify simvastatin nano structured lipid carrier, by: simvastatin, matrix material, polyethylene glycol mono stearate, the polyglycol distearate of mono amino terminal, N, N ' two succinimidyl carbonate and ICAM 1 monoclonal antibody formed.The present invention is with simvastatin as medicine, simvastatin nano structured lipid carrier is prepared with solvent diffusion method, by its surface being carried out the modification of ICAM 1 monoclonal antibody, build the ICAM 1 monoclonal antibody simvastatin nano structured lipid carrier of selectively targeted ALI pulmonary vascular endothelial cell.The lipid carrier of the present invention can targeting ALI pulmonary vascular endothelial cell, delay the release of simvastatin, improve the drug accumulation amount of pulmonary lesions thus improve Drug therapy efficiency, reduce the liver toxicity of simvastatin, muscle poison.From the pathogenesis reversing the damage of ALI pulmonary vascular endothelial cell, it is effectively improved ALI disease.

Description

ICAM-1 monoclonal antibody modifies simvastatin nano structured lipid carrier and preparation and application
Technical field
The invention belongs to the preparation of medicine Novel Drug Delivery Systems, be specially the ICAM-1 receptor to vascular endothelial cell and have The ICAM-1 monoclonal antibody of specific binding capacity modifies simvastatin nano structured lipid carrier and preparation method thereof, and Its application in prepared by the medicine of acute lung injury.
Background technology
Acute lung injury (acute lung injury, ALI) is a kind of common clinical syndrome, and its advanced stage is Adult respiratory distress syndrome (acute respiratory distress syndrome, ARDS), has higher sickness rate And mortality rate, the life and health of the mankind in serious threat up to now.The pathology of ALI characterizes and is presented as that a large amount of diffusivity inflammation is thin In born of the same parents' lung, infiltration, alveolar and pulmonary parenchyma inner height edema, alveolar hyaline membrane are formed and the interstitial pulmonary fibrosis in later stage.In the past Ten years, the improvement for ALI is still based primarily upon several Supporting Therapys technology, such as small tidal volume ventilation, gives PEEP, lung Liquid management, antimicrobial therapy etc..The most a series of pharmacotherapy is also applied to the improvement of ALI, such as vasodilation The Inhalation in Treating of (nitric oxide, prostacyclin etc.), Adrenal Glucocorticoid, ketoconazole, antioxidant etc., but there is no so far Considerable clinical therapeutic efficacy.This make us not satisfied Drug therapy result, be attributable to induce ALI to a certain extent sick Various heterogeneity of cause and the pathophysiological change that alveolar is lasting during ALI disease is sent out.Therefore from the complicated morbidity of ALI Mechanism is set out, and solves the basic reason of ALI morbidity, thus seeks a new ALI therapy approach and the most more and more closed Note.
Relative to the pathogeny such as the most excessive cascade of response of inflammation, oxidative stress is unbalance, Abnormality of Coagulation Fibrinolysis, lung blood Pipe damage is widely regarded as ALI disease and is sent out leading reason the most originally.Studies have reported that the blood vessel endothelium caused by ALI changes ALI morbidity plays important regulating and controlling effect.And neutrophilic granulocyte Dependent is widely regarded as the main way of lung endothelial injury Footpath.Accumulate the neutrophilic granulocyte in pulmonary microvasculature to activate accordingly with it due to inherent immunity, by retting conditions and Release Pro-inflammatory Cytokine, reactive oxygen species, protease and coagulant pigr molecule etc. cause endothelial cell damage, pulmonary vascular permeability Strengthen, and then induction pulmonary edema occurs.Lung Endothelial Dysfunction can further result in inflammatory cell in lung adhere to and migrate strengthen, Inflammatory factor and the vicious cycle of coagulant anticoagulant Imbalance.Additionally, based on pulmonary vascular endothelial cell as regulation pulmonary and complete The main metabolic organ of body blood vessel balance, so the damage from reversing pulmonary vascular endotheliocyte is started with, finds one and effectively controls The means treating ALI have highly important researching value and clinical meaning.
Statins is as the traditional Hydroxymethylglutaryl CoA-reductase inhibitors for reducing blood fat, existing Stage is the most more and more applied in the research that ALI protects.This multidirectional medicinal property being had based on it, including Protect vascular endothelial cell cytoskeleton, the generation of reduction superoxides, promote NO synthesis, rise endothelial cell inflammation decay thing Matter integrin-the β 4 and claudin-5 relevant to intercellular tight junction.Before the clinic that ALI is protected by statins Research makes remarkable progress, equally including changing the ALI that the modes such as intratracheal instillation LPS, mechanical ventilation, radiation cause Kind, and its common starting stage pneumonia disease is served positive curative effect of medication equally.Although Statins up to now Medicine is applied to the treatment of ALI the most clinically, but the improvement of human body ALI is studied and achieved actively by recent years some Achievement.Have research display, carry out the patient of statins prevention or treatment in advance, can significantly reduce ICU patient be in hospital front 4 It sepsis and the incidence rate of ALI/ARDS, fully demonstrate statins for preventing or treating the potential of acute lung injury Clinical value, so that statins is likely to become treats ALI's by reversing pulmonary vascular endothelial cell damage process A new generation's medicine.But Statins was still needed before really becoming the medicine of ALI and is overcome following point: 1. uncertain therapeutic efficacy Fixed, the randomized control study carried out in Ireland for 2010 shows: 80mg simvastatin is used for treating ALI mechanical ventilation and suffers from Person, although the 14th day treatment group patient has more preferable oxygenation index and ventilation parameters, but poor without significance compared with blank group Different；2. untoward reaction is many, mainly shows as sarcolysis and the liver poison caused due to oxidative stress and mitochondrial dysfunction Property, central nervous system toxicity etc..Based on fat-soluble statins can by the way of Passive diffusion by cellular uptake thus It is easier to act on cells play drug effect, therefore the present invention selects the simvastatin that lipotropy in statins is the strongest to be research medicine Thing.But, also due to higher tissue permeability, for water solublity statin, fat-soluble statin has higher medicine Thing incidence of side effects.For overcoming statins for treating the above-mentioned deficiency of ALI, statins is wrapped in by this patent In nano structured lipid carrier, by nano structured lipid carrier by targeted drug delivery to ALI vascular endothelial cell, strengthen disease The drug accumulation at stove position, significantly improves Drug therapy efficiency.Damage reversing vascular endothelial cell from the pathogenesis of ALI While wound, reduce the untoward reaction of statins.
Research report ALI can induction of vascular endothelial cell produce different kind organism label, including von Willebrand because of Son, Ang-2 and intercellular adhesion molecule-1 (ICAM-1).ICAM-1 is as the transmembrane glycopeptide being expressed in endothelial cell surface In vain, its expression is closely related with inflammatory intensity.External multidigit scholar is applied in the targeting design of nano-carrier, he Achievement in research confirm: by nano-carrier surface being carried out the modification of ICAM-1 monoclonal antibody, utilize ICAM-1 monoclonal antibody With the principle that Surface of Vascular Endothelial Cells ICAM-1 receptor-specific is combined, the nano-carrier target to vascular endothelial cell can be realized To conveying.The present invention selects the nano structured lipid carrier in administration nano-drug administration system as drug delivery system.Nanostructured fat Matter carrier, as a kind of novel drug-supplying system, has a following delivery properties: the best biocompatibility；2. medicine delays, controls Release；The most relatively low bio-toxicity；The most easily prepared etc..Above-mentioned characteristic based on nano structured lipid carrier, by right The efficient packet of statins carries, and nano-carrier surface carries out ICAM-1 monoclonal antibody modification, makes statins targeted Imagination to ALI vascular endothelial cell becomes a reality.
Summary of the invention
It is an object of the present invention to provide a kind of ICAM-1 monoclonal antibody and modify simvastatin nano structured lipid carrier.Should ICAM-1 monoclonal antibody modify simvastatin nano structured lipid carrier mainly by: simvastatin, matrix material, polyethyleneglycol are stearic Acid esters (MW 2000), the polyglycol distearate of mono amino terminal, N, N '-two succinimidyl carbonate and ICAM- 1 monoclonal antibody is formed.Formulation ingredients mass percent is 4.98%-14.95% simvastatin, 83.72%-93.68% Matrix material, the polyethylene glycol mono stearate (MW 2000) of 0%-9.97%, the poly-second two of mono amino terminal of 1.00% Alcohol stearate, 0.22%N, N '-two succinimidyl carbonate and the ICAM-1 monoclonal antibody of 0.11%.Described fat Material selects one or both in glyceryl monostearate, stearic acid, oleic acid, medium chain length fatty acid triglyceride.
Second object of the present invention is to provide ICAM-1 monoclonal antibody and modifies the preparation of simvastatin nano structured lipid carrier Method, realizes especially by below scheme:
1. synthesize the polyglycol distearate of mono amino terminal
Weigh 119.8mg 1-(3-dimethylamino-propyl)-3-ethyl-carbodiimide hydrochloride, 24.0mg N-hydroxysuccinimidyl Acid imide, 35.5mg stearic acid is dissolved in 3ml anhydrous dimethyl sulphoxide, reacts 30min, by reactant liquor under the conditions of 400rpm 60 DEG C Being slowly added dropwise to the double N-terminal polyglycol solution of 250mg of 2ml anhydrous dimethyl sulphoxide dissolving, under room temperature, 400rpm stirs Mix reaction 24h.After reaction terminates, reactant liquor is transferred in bag filter (MWCO:7kDa), pure water dialysis 48h, removes water solublity By-product, lyophilization i.e. obtains the polyglycol distearate of mono amino terminal.
The preparation of the simvastatin nano structured lipid carrier that 2.ICAM-1 monoclonal antibody is modified
Solvent diffusion method prepares simvastatin nano structured lipid carrier, concrete grammar: precision weighs the pungent of 5-15mg and cuts down Statin, the matrix material of 84-94mg, the polyglycol distearate (MW 2000) of 0-10mg and the N-terminal of 1mg synthesis Polyethylene glycol mono stearate, altogether 100mg, add 1ml ethanol and dissolve as organic facies, water-bath 60 DEG C heating, make material complete Full thawing.The material that syringe absorption 0.45ml melts, ethanol dilution to 0.5ml, with deionized water as disperse medium, In the deionized water of the 4.5ml that under 400rpm mixing speed, organic facies is rapidly injected successively 60 DEG C of heating in water bath, inject complete Take out after stirring 30s, naturally cool to room temperature, prepare the simvastatin nano structured lipid carrier of lipid concentration 9mg/ml.Add Enter the N of 0.1mg, N '-two succinimidyl carbonate, vortex 1min be uniformly dispersed after incubation reaction 3h under room temperature, add 50 μ G ICAM-1 monoclonal antibody, vortex 1min continues incubation reaction 3h under room temperature after being uniformly dispersed.This reaction is by N, N '-two amber Amber imide carbonic ester connects the amino of Polyethylene Glycol and the amino in ICAM-1 monoclonal antibody on pharmaceutical carrier, by antibody Successfully being grafted onto on pharmaceutical carrier, lyophilization i.e. obtains ICAM-1 monoclonal antibody and modifies simvastatin nano structured lipid carrier.
It is also another object of the present invention to provide described ICAM-1 monoclonal antibody and modify simvastatin nano structured lipid carrier Application in preparing targeting ALI pulmonary vascular endothelial cell medicine.
The present invention is with simvastatin as medicine, with matrix material and polyethylene glycol mono stearate as main material, with water Property solvent diffusion method prepares simvastatin nano structured lipid carrier, repaiies by its surface is carried out ICAM-1 monoclonal antibody Decorations, build the ICAM-1 monoclonal antibody simvastatin nano structured lipid carrier of selectively targeted ALI pulmonary vascular endothelial cell, and evaluate Its improvement result to ALI.From the pathogenesis reversing the damage of ALI pulmonary vascular endothelial cell, seek a kind of safe efficient ALI statin targeted therapy new way.
Usefulness of the present invention, the ICAM-1 monoclonal antibody simvastatin nano structured lipid carrier being provided can targeting ALI pulmonary vascular endothelial cell, delay the release of simvastatin, improve the drug accumulation amount of pulmonary lesions thus improve Drug therapy Efficiency, reduces the liver toxicity of simvastatin, muscle poison.From the pathogenesis reversing the damage of ALI pulmonary vascular endothelial cell, have Effect improves ALI disease.
Accompanying drawing explanation
Fig. 1 is simvastatin nano structured lipid carrier transmission electron microscope photo.
Fig. 2 is simvastatin nano structured lipid carrier drug release profiles.
Fig. 3 is simvastatin nano structured lipid carrier distribution in vivo living imaging.
Fig. 4 is the protective effect to acute lung injury of the simvastatin nano structured lipid carrier.
Detailed description of the invention
The present invention is further described in conjunction with the accompanying drawings and embodiments.
Embodiment 1
1. synthesize the polyglycol distearate of mono amino terminal
Weigh 119.8mg 1-(3-dimethylamino-propyl)-3-ethyl-carbodiimide hydrochloride, 24.0mg N-hydroxysuccinimidyl Acid imide, 35.5mg stearic acid is dissolved in 3ml anhydrous dimethyl sulphoxide, reacts 30min, by reactant liquor under the conditions of 400rpm 60 DEG C Being slowly added dropwise to the double N-terminal polyglycol solution of 250mg of 2ml anhydrous dimethyl sulphoxide dissolving, under room temperature, 400rpm stirs Mix reaction 24h.After reaction terminates, reactant liquor is transferred in bag filter (MWCO:7kDa), pure water dialysis 48h, removes water solublity By-product, lyophilization i.e. obtains the polyglycol distearate of mono amino terminal.
The preparation of the simvastatin nano structured lipid carrier that 2.ICAM-1 monoclonal antibody is modified
Solvent diffusion method prepares simvastatin nano structured lipid carrier, concrete grammar: precision weigh 5mg simvastatin, 94mg glyceryl monostearate, the polyglycol distearate of the mono amino terminal of 1mg synthesis, altogether 100mg, add 1ml ethanol Dissolve as organic facies, water-bath 60 DEG C heating, make material melt completely.Syringe draws the material that 0.45ml melts, and ethanol is dilute Release to 0.5ml, with deionized water as disperse medium, organic facies is rapidly injected successively 60 DEG C of water-baths under 400rpm mixing speed In the deionized water of the 4.5ml of heating, take out after injecting complete stirring 30s, naturally cool to room temperature, prepare lipid concentration 9mg/ The simvastatin nano structured lipid carrier of ml.Adding N, N '-two succinimidyl carbonate of 0.1mg, vortex 1min divides After dissipating uniformly, incubation reaction 3h under room temperature, adds 50 μ g ICAM-1 monoclonal antibodies, and vortex 1min continues under room temperature after being uniformly dispersed Continuous incubation reaction 3h.This reaction by N, N '-two succinimidyl carbonate connect the amino of Polyethylene Glycol on pharmaceutical carrier With the amino in ICAM-1 monoclonal antibody, being successfully grafted onto on pharmaceutical carrier by antibody, lyophilization i.e. obtains ICAM-1 monoclonal antibody The simvastatin nano structured lipid carrier modified.
Use the size of nanoparticle prepared by dynamic light scattering determination.Sample is diluted to deionized water 0.1mg/ml, measures pungent cutting down before and after ICAM-1 monoclonal antibody is modified with Malvern Zetasizer Nano-S90 nano-particle size analysis instrument The particle diameter of statin nano structured lipid carrier is respectively 204.5 ± 4.56nm, 217.4 ± 24.25nm.
Acid adjustment centrifuging is used to coordinate envelop rate and the load of Syrups by HPLC simvastatin nano structured lipid carrier Dose.Stand to add to the simvastatin nanoparticle dispersion liquid without the modification of ICAM-1 monoclonal antibody of room temperature after preparation and fit The hydrochloric acid of amount 0.1M, the PH to system is down to 1.2 to promote nanoparticle flocculate, and 20000rpm is centrifuged 30min, HPLC mensuration supernatant Liquid drug concentration, calculates envelop rate and the drug loading of simvastatin nano structured lipid carrier as follows.
Envelop rate=(drug quality in lipid nanoparticle/total dosage) × 100%
Drug loading=(drug quality in lipid nanoparticle/drug-carrying nanometer particle gross mass) × 100%
After measured, envelop rate and the drug loading of the simvastatin nano structured lipid carrier modified without ICAM-1 monoclonal antibody divide It is not 81.32% and 4.10%.
The preparation of the simvastatin nano structured lipid carrier that embodiment 2 ICAM-1 monoclonal antibody is modified
Solvent diffusion method prepares simvastatin nano structured lipid carrier, concrete grammar: precision weigh 10mg simvastatin, 89mg glyceryl monostearate, the polyglycol distearate of the mono amino terminal of 1mg synthesis, altogether 100mg, add 1ml ethanol Dissolve as organic facies, water-bath 60 DEG C heating, make material melt completely.Syringe draws the material that 0.45ml melts, and ethanol is dilute Release to 0.5ml, with deionized water as disperse medium, organic facies is rapidly injected successively 60 DEG C of water-baths under 400rpm mixing speed In the deionized water of the 4.5ml of heating, take out after injecting complete stirring 30s, naturally cool to room temperature, prepare lipid concentration 9mg/ The simvastatin nano structured lipid carrier of ml.Adding N, N '-two succinimidyl carbonate of 0.1mg, vortex 1min divides After dissipating uniformly, incubation reaction 3h under room temperature, adds 50 μ g ICAM-1 monoclonal antibodies, and vortex 1min continues under room temperature after being uniformly dispersed Continuous incubation reaction 3h.This reaction by N, N '-two succinimidyl carbonate connect the amino of Polyethylene Glycol on pharmaceutical carrier With the amino in ICAM-1 monoclonal antibody, being successfully grafted onto on pharmaceutical carrier by antibody, lyophilization i.e. obtains ICAM-1 monoclonal antibody The simvastatin nano structured lipid carrier modified.
Use the size of nanoparticle prepared by dynamic light scattering determination.Sample is diluted to deionized water 0.1mg/ml, measures pungent cutting down before and after ICAM-1 monoclonal antibody is modified with Malvern Zetasizer Nano-S90 nano-particle size analysis instrument The particle diameter of statin nano structured lipid carrier is respectively 245.6 ± 12.4nm, 257.2 ± 20.7nm.
Acid adjustment centrifuging is used to coordinate envelop rate and the load of Syrups by HPLC simvastatin nano structured lipid carrier Dose.Stand to add to the simvastatin nanoparticle dispersion liquid without the modification of ICAM-1 monoclonal antibody of room temperature after preparation and fit The hydrochloric acid of amount 0.1M, the PH to system is down to 1.2 to promote nanoparticle flocculate, and 20000rpm is centrifuged 30min, HPLC mensuration supernatant Liquid drug concentration, calculates envelop rate and the drug loading of simvastatin nano structured lipid carrier as follows.
Envelop rate=(drug quality in lipid nanoparticle/total dosage) × 100%
Drug loading=(drug quality in lipid nanoparticle/drug-carrying nanometer particle gross mass) × 100%
After measured, envelop rate and the drug loading of the simvastatin nano structured lipid carrier modified without ICAM-1 monoclonal antibody divide It is not 85.70% and 8.69%.
The preparation of the simvastatin nano structured lipid carrier that embodiment 3 ICAM-1 monoclonal antibody is modified
Solvent diffusion method prepares simvastatin nano structured lipid carrier, concrete grammar: precision weigh 15mg simvastatin, 84mg glyceryl monostearate, the polyglycol distearate of the mono amino terminal of 1mg synthesis, altogether 100mg, add 1ml ethanol Dissolve as organic facies, water-bath 60 DEG C heating, make material melt completely.Syringe draws the material that 0.45ml melts, and ethanol is dilute Release to 0.5ml, with deionized water as disperse medium, organic facies is rapidly injected successively 60 DEG C of water-baths under 400rpm mixing speed In the deionized water of the 4.5ml of heating, take out after injecting complete stirring 30s, naturally cool to room temperature, prepare lipid concentration 9mg/ The simvastatin nano structured lipid carrier of ml.Adding N, N '-two succinimidyl carbonate of 0.1mg, vortex 1min divides After dissipating uniformly, incubation reaction 3h under room temperature, adds 50 μ g ICAM-1 monoclonal antibodies, and vortex 1min continues under room temperature after being uniformly dispersed Continuous incubation reaction 3h.This reaction by N, N '-two succinimidyl carbonate connect the amino of Polyethylene Glycol on pharmaceutical carrier With the amino in ICAM-1 monoclonal antibody, being successfully grafted onto on pharmaceutical carrier by antibody, lyophilization i.e. obtains ICAM-1 monoclonal antibody The simvastatin nano structured lipid carrier modified.
Use the size of nanoparticle prepared by dynamic light scattering determination.Sample is diluted to deionized water 0.1mg/ml, measures pungent cutting down before and after ICAM-1 monoclonal antibody is modified with Malvern Zetasizer Nano-S90 nano-particle size analysis instrument The particle diameter of statin nano structured lipid carrier is respectively 274.3 ± 7.3nm, 278.9 ± 13.6nm.
Acid adjustment centrifuging is used to coordinate envelop rate and the load of Syrups by HPLC simvastatin nano structured lipid carrier Dose.Stand to add to the simvastatin nanoparticle dispersion liquid without the modification of ICAM-1 monoclonal antibody of room temperature after preparation and fit The hydrochloric acid of amount 0.1M, the PH to system is down to 1.2 to promote nanoparticle flocculate, and 20000rpm is centrifuged 30min, HPLC mensuration supernatant Liquid drug concentration, calculates envelop rate and the drug loading of simvastatin nano structured lipid carrier as follows.
Envelop rate=(drug quality in lipid nanoparticle/total dosage) × 100%
Drug loading=(drug quality in lipid nanoparticle/drug-carrying nanometer particle gross mass) × 100%
After measured, envelop rate and the drug loading of the simvastatin nano structured lipid carrier modified without ICAM-1 monoclonal antibody divide It is not 75.42% and 11.75%.
The preparation of the simvastatin nano structured lipid carrier that embodiment 4 ICAM-1 monoclonal antibody is modified
Solvent diffusion method prepares simvastatin nano structured lipid carrier, concrete grammar: precision weigh 5mg simvastatin, 94mg stearic acid, the polyglycol distearate of the mono amino terminal of 1mg synthesis, altogether 100mg, add 1ml ethanol and dissolve conduct Organic facies, water-bath 60 DEG C heating, make material melt completely.Syringe draws the material that 0.45ml melts, and ethanol dilution is extremely 0.5ml, with deionized water as disperse medium, is rapidly injected 60 DEG C of heating in water bath successively under 400rpm mixing speed by organic facies 4.5ml deionized water in, inject and take out after complete stirring 30s, naturally cool to room temperature, prepare lipid concentration 9mg/ml's Simvastatin nano structured lipid carrier.Adding N, N '-two succinimidyl carbonate of 0.1mg, vortex 1min dispersion is all Incubation reaction 3h under even rear room temperature, adds 50 μ g ICAM-1 monoclonal antibodies, and vortex 1min continues to incubate under room temperature after being uniformly dispersed Educate reaction 3h.This reaction by N, N '-two succinimidyl carbonate connect on pharmaceutical carrier the amino of Polyethylene Glycol with Amino in ICAM-1 monoclonal antibody, is successfully grafted onto antibody on pharmaceutical carrier, and lyophilization i.e. obtains ICAM-1 monoclonal antibody and repaiies The simvastatin nano structured lipid carrier of decorations.
Use the size of nanoparticle prepared by dynamic light scattering determination.Sample is diluted to deionized water 0.1mg/ml, measures pungent cutting down before and after ICAM-1 monoclonal antibody is modified with Malvern Zetasizer Nano-S90 nano-particle size analysis instrument The particle diameter of statin nano structured lipid carrier is respectively 213.7 ± 10.5nm, 223.9 ± 7.7nm.
Acid adjustment centrifuging is used to coordinate envelop rate and the load of Syrups by HPLC simvastatin nano structured lipid carrier Dose.Stand to add to the simvastatin nanoparticle dispersion liquid without the modification of ICAM-1 monoclonal antibody of room temperature after preparation and fit The hydrochloric acid of amount 0.1M, the PH to system is down to 1.2 to promote nanoparticle flocculate, and 20000rpm is centrifuged 30min, HPLC mensuration supernatant Liquid drug concentration, calculates envelop rate and the drug loading of simvastatin nano structured lipid carrier as follows.
Envelop rate=(drug quality in lipid nanoparticle/total dosage) × 100%
Drug loading=(drug quality in lipid nanoparticle/drug-carrying nanometer particle gross mass) × 100%
After measured, envelop rate and the drug loading of the simvastatin nano structured lipid carrier modified without ICAM-1 monoclonal antibody divide It is not 79.53% and 4.02%.
The preparation of the simvastatin nano structured lipid carrier that embodiment 5 ICAM-1 monoclonal antibody is modified
Solvent diffusion method prepares simvastatin nano structured lipid carrier, concrete grammar: precision weigh 10mg simvastatin, 89mg stearic acid, the polyglycol distearate of the mono amino terminal of 1mg synthesis, altogether 100mg, add 1ml ethanol and dissolve conduct Organic facies, water-bath 60 DEG C heating, make material melt completely.Syringe draws the material that 0.45ml melts, and ethanol dilution is extremely 0.5ml, with deionized water as disperse medium, is rapidly injected 60 DEG C of heating in water bath successively under 400rpm mixing speed by organic facies 4.5ml deionized water in, inject and take out after complete stirring 30s, naturally cool to room temperature, prepare lipid concentration 9mg/ml's Simvastatin nano structured lipid carrier.Adding N, N '-two succinimidyl carbonate of 0.1mg, vortex 1min dispersion is all Incubation reaction 3h under even rear room temperature, adds 50 μ g ICAM-1 monoclonal antibodies, and vortex 1min continues to incubate under room temperature after being uniformly dispersed Educate reaction 3h.This reaction by N, N '-two succinimidyl carbonate connect on pharmaceutical carrier the amino of Polyethylene Glycol with Amino in ICAM-1 monoclonal antibody, is successfully grafted onto antibody on pharmaceutical carrier, and lyophilization i.e. obtains ICAM-1 monoclonal antibody and repaiies The simvastatin nano structured lipid carrier of decorations.
Use the size of nanoparticle prepared by dynamic light scattering determination.Sample is diluted to deionized water 0.1mg/ml, measures pungent cutting down before and after ICAM-1 monoclonal antibody is modified with Malvern Zetasizer Nano-S90 nano-particle size analysis instrument The particle diameter of statin nano structured lipid carrier is respectively 252.7 ± 13.6nm, 277.89 ± 11.5nm.
Acid adjustment centrifuging is used to coordinate envelop rate and the load of Syrups by HPLC simvastatin nano structured lipid carrier Dose.Stand to add to the simvastatin nanoparticle dispersion liquid without the modification of ICAM-1 monoclonal antibody of room temperature after preparation and fit The hydrochloric acid of amount 0.1M, the PH to system is down to 1.2 to promote nanoparticle flocculate, and 20000rpm is centrifuged 30min, HPLC mensuration supernatant Liquid drug concentration, calculates envelop rate and the drug loading of simvastatin nano structured lipid carrier as follows.
Envelop rate=(drug quality in lipid nanoparticle/total dosage) × 100%
Drug loading=(drug quality in lipid nanoparticle/drug-carrying nanometer particle gross mass) × 100%
After measured, envelop rate and the drug loading of the simvastatin nano structured lipid carrier modified without ICAM-1 monoclonal antibody divide It is not 83.11% and 8.45%.
The preparation of the simvastatin nano structured lipid carrier that embodiment 6 ICAM-1 monoclonal antibody is modified
Solvent diffusion method prepares simvastatin nano structured lipid carrier, concrete grammar: precision weigh 15mg simvastatin, 84mg stearic acid, the polyglycol distearate of the mono amino terminal of 1mg synthesis, altogether 100mg, add 1ml ethanol and dissolve conduct Organic facies, water-bath 60 DEG C heating, make material melt completely.Syringe draws the material that 0.45ml melts, and ethanol dilution is extremely 0.5ml, with deionized water as disperse medium, is rapidly injected 60 DEG C of heating in water bath successively under 400rpm mixing speed by organic facies 4.5ml deionized water in, inject and take out after complete stirring 30s, naturally cool to room temperature, prepare lipid concentration 9mg/ml's Simvastatin nano structured lipid carrier.Adding N, N '-two succinimidyl carbonate of 0.1mg, vortex 1min dispersion is all Incubation reaction 3h under even rear room temperature, adds 50 μ g ICAM-1 monoclonal antibodies, and vortex 1min continues to incubate under room temperature after being uniformly dispersed Educate reaction 3h.This reaction by N, N '-two succinimidyl carbonate connect on pharmaceutical carrier the amino of Polyethylene Glycol with Amino in ICAM-1 monoclonal antibody, is successfully grafted onto antibody on pharmaceutical carrier, and lyophilization i.e. obtains ICAM-1 monoclonal antibody and repaiies The simvastatin nano structured lipid carrier of decorations.
Use the size of nanoparticle prepared by dynamic light scattering determination.Sample is diluted to deionized water 0.1mg/ml, measures pungent cutting down before and after ICAM-1 monoclonal antibody is modified with Malvern Zetasizer Nano-S90 nano-particle size analysis instrument The particle diameter of statin nano structured lipid carrier is respectively 280.2 ± 9.3nm, 283.5 ± 3.5nm.
Acid adjustment centrifuging is used to coordinate envelop rate and the load of Syrups by HPLC simvastatin nano structured lipid carrier Dose.Stand to add to the simvastatin nanoparticle dispersion liquid without the modification of ICAM-1 monoclonal antibody of room temperature after preparation and fit The hydrochloric acid of amount 0.1M, the PH to system is down to 1.2 to promote nanoparticle flocculate, and 20000rpm is centrifuged 30min, HPLC mensuration supernatant Liquid drug concentration, calculates envelop rate and the drug loading of simvastatin nano structured lipid carrier as follows.
Envelop rate=(drug quality in lipid nanoparticle/total dosage) × 100%
Drug loading=(drug quality in lipid nanoparticle/drug-carrying nanometer particle gross mass) × 100%
After measured, envelop rate and the drug loading of the simvastatin nano structured lipid carrier modified without ICAM-1 monoclonal antibody divide It is not 74.25% and 11.58%.
The preparation of the simvastatin nano structured lipid carrier that embodiment 7 ICAM-1 monoclonal antibody is modified
Solvent diffusion method prepares simvastatin nano structured lipid carrier, concrete grammar: precision weigh 5mg simvastatin, 84mg glyceryl monostearate, 10mg oleic acid, the polyglycol distearate of the mono amino terminal of 1mg synthesis, 100mg, adds altogether Enter 1ml ethanol to dissolve as organic facies, water-bath 60 DEG C heating, make material melt completely.Syringe draws the material that 0.45ml melts Material, ethanol dilution to 0.5ml, with deionized water as disperse medium, under 400rpm mixing speed, organic facies is the most quickly noted In the deionized water of the 4.5ml entering 60 DEG C of heating in water bath, take out after injecting complete stirring 30s, naturally cool to room temperature, prepare fat The simvastatin nano structured lipid carrier of matter concentration 9mg/ml.Add N, N '-two succinimidyl carbonate of 0.1mg, Vortex 1min be uniformly dispersed after incubation reaction 3h under room temperature, add 50 μ g ICAM-1 monoclonal antibodies, vortex 1min is uniformly dispersed Incubation reaction 3h is continued under rear room temperature.This reaction by N, N '-two succinimidyl carbonate connect poly-second on pharmaceutical carrier Amino in the amino of glycol and ICAM-1 monoclonal antibody, is successfully grafted onto antibody on pharmaceutical carrier, lyophilization and get final product The simvastatin nano structured lipid carrier that ICAM-1 monoclonal antibody is modified.
Use the size of nanoparticle prepared by dynamic light scattering determination.Sample is diluted to deionized water 0.1mg/ml, measures pungent cutting down before and after ICAM-1 monoclonal antibody is modified with Malvern Zetasizer Nano-S90 nano-particle size analysis instrument The particle diameter of statin nano structured lipid carrier is respectively 284.5 ± 5.7nm, 290.4 ± 14.2nm.
Acid adjustment centrifuging is used to coordinate envelop rate and the load of Syrups by HPLC simvastatin nano structured lipid carrier Dose.Stand to add to the simvastatin nanoparticle dispersion liquid without the modification of ICAM-1 monoclonal antibody of room temperature after preparation and fit The hydrochloric acid of amount 0.1M, the PH to system is down to 1.2 to promote nanoparticle flocculate, and 20000rpm is centrifuged 30min, HPLC mensuration supernatant Liquid drug concentration, calculates envelop rate and the drug loading of simvastatin nano structured lipid carrier as follows.
Envelop rate=(drug quality in lipid nanoparticle/total dosage) × 100%
Drug loading=(drug quality in lipid nanoparticle/drug-carrying nanometer particle gross mass) × 100%
After measured, envelop rate and the drug loading of the simvastatin nano structured lipid carrier modified without ICAM-1 monoclonal antibody divide It is not 83.53% and 4.21%.
The preparation of the simvastatin nano structured lipid carrier that embodiment 8 ICAM-1 monoclonal antibody is modified
Solvent diffusion method prepares simvastatin nano structured lipid carrier, concrete grammar: precision weigh 5mg simvastatin, 74mg glyceryl monostearate, 20mg oleic acid, the polyglycol distearate of the mono amino terminal of 1mg synthesis, 100mg, adds altogether Enter 1ml ethanol to dissolve as organic facies, water-bath 60 DEG C heating, make material melt completely.Syringe draws the material that 0.45ml melts Material, ethanol dilution to 0.5ml, with deionized water as disperse medium, under 400rpm mixing speed, organic facies is the most quickly noted In the deionized water of the 4.5ml entering 60 DEG C of heating in water bath, take out after injecting complete stirring 30s, naturally cool to room temperature, prepare fat The simvastatin nano structured lipid carrier of matter concentration 9mg/ml.Add N, N '-two succinimidyl carbonate of 0.1mg, Vortex 1min be uniformly dispersed after incubation reaction 3h under room temperature, add 50 μ g ICAM-1 monoclonal antibodies, vortex 1min is uniformly dispersed Incubation reaction 3h is continued under rear room temperature.This reaction by N, N '-two succinimidyl carbonate connect poly-second on pharmaceutical carrier Amino in the amino of glycol and ICAM-1 monoclonal antibody, is successfully grafted onto antibody on pharmaceutical carrier, lyophilization and get final product The simvastatin nano structured lipid carrier that ICAM-1 monoclonal antibody is modified.
Use the size of nanoparticle prepared by dynamic light scattering determination.Sample is diluted to deionized water 0.1mg/ml, measures pungent cutting down before and after ICAM-1 monoclonal antibody is modified with Malvern Zetasizer Nano-S90 nano-particle size analysis instrument The particle diameter of statin nano structured lipid carrier is respectively 312.5 ± 14.3nm, 316.7 ± 17.1nm.
Acid adjustment centrifuging is used to coordinate envelop rate and the load of Syrups by HPLC simvastatin nano structured lipid carrier Dose.Stand to add to the simvastatin nanoparticle dispersion liquid without the modification of ICAM-1 monoclonal antibody of room temperature after preparation and fit The hydrochloric acid of amount 0.1M, the PH to system is down to 1.2 to promote nanoparticle flocculate, and 20000rpm is centrifuged 30min, HPLC mensuration supernatant Liquid drug concentration, calculates envelop rate and the drug loading of simvastatin nano structured lipid carrier as follows.
Envelop rate=(drug quality in lipid nanoparticle/total dosage) × 100%
Drug loading=(drug quality in lipid nanoparticle/drug-carrying nanometer particle gross mass) × 100%
After measured, envelop rate and the drug loading of the simvastatin nano structured lipid carrier modified without ICAM-1 monoclonal antibody divide It is not 87.45% and 4.40%.
The preparation of the simvastatin nano structured lipid carrier that embodiment 9 ICAM-1 monoclonal antibody is modified
Solvent diffusion method prepares simvastatin nano structured lipid carrier, concrete grammar: precision weigh 5mg simvastatin, 64mg glyceryl monostearate, 30mg oleic acid, the polyglycol distearate of the mono amino terminal of 1mg synthesis, 100mg, adds altogether Enter 1ml ethanol to dissolve as organic facies, water-bath 60 DEG C heating, make material melt completely.Syringe draws the material that 0.45ml melts Material, ethanol dilution to 0.5ml, with deionized water as disperse medium, under 400rpm mixing speed, organic facies is the most quickly noted In the deionized water of the 4.5ml entering 60 DEG C of heating in water bath, take out after injecting complete stirring 30s, naturally cool to room temperature, prepare fat The simvastatin nano structured lipid carrier of matter concentration 9mg/ml.Add N, N '-two succinimidyl carbonate of 0.1mg, Vortex 1min be uniformly dispersed after incubation reaction 3h under room temperature, add 50 μ g ICAM-1 monoclonal antibodies, vortex 1min is uniformly dispersed Incubation reaction 3h is continued under rear room temperature.This reaction by N, N '-two succinimidyl carbonate connect poly-second on pharmaceutical carrier Amino in the amino of glycol and ICAM-1 monoclonal antibody, is successfully grafted onto antibody on pharmaceutical carrier, lyophilization and get final product The simvastatin nano structured lipid carrier that ICAM-1 monoclonal antibody is modified.
Use the size of nanoparticle prepared by dynamic light scattering determination.Sample is diluted to deionized water 0.1mg/ml, measures pungent cutting down before and after ICAM-1 monoclonal antibody is modified with Malvern Zetasizer Nano-S90 nano-particle size analysis instrument The particle diameter of statin nano structured lipid carrier is respectively 342.6 ± 24.6nm, 347.8 ± 19.2nm.
Acid adjustment centrifuging is used to coordinate envelop rate and the load of Syrups by HPLC simvastatin nano structured lipid carrier Dose.Stand to add to the simvastatin nanoparticle dispersion liquid without the modification of ICAM-1 monoclonal antibody of room temperature after preparation and fit The hydrochloric acid of amount 0.1M, the PH to system is down to 1.2 to promote nanoparticle flocculate, and 20000rpm is centrifuged 30min, HPLC mensuration supernatant Liquid drug concentration, calculates envelop rate and the drug loading of simvastatin nano structured lipid carrier as follows.
Envelop rate=(drug quality in lipid nanoparticle/total dosage) × 100%
Drug loading=(drug quality in lipid nanoparticle/drug-carrying nanometer particle gross mass) × 100%
After measured, envelop rate and the drug loading of the simvastatin nano structured lipid carrier modified without ICAM-1 monoclonal antibody divide It is not 85.32% and 4.30%.
The preparation of the simvastatin nano structured lipid carrier that embodiment 10 ICAM-1 monoclonal antibody is modified
Solvent diffusion method prepares simvastatin nano structured lipid carrier, concrete grammar: precision weigh 5mg simvastatin, 84mg glyceryl monostearate, 10mg medium chain length fatty acid triglyceride, the Polyethylene Glycol stearic acid of the mono amino terminal of 1mg synthesis Ester, altogether 100mg, add 1ml ethanol and dissolve as organic facies, water-bath 60 DEG C heating, make material melt completely.Syringe is drawn The material that 0.45ml melts, ethanol dilution to 0.5ml, with deionized water as disperse medium, will have under 400rpm mixing speed In the deionized water of the 4.5ml that machine is rapidly injected 60 DEG C of heating in water bath mutually successively, take out after injecting complete stirring 30s, the coldest But to room temperature, the simvastatin nano structured lipid carrier of lipid concentration 9mg/ml is prepared.Add N, N '-two succinum of 0.1mg Imide carbonic ester, vortex 1min be uniformly dispersed after incubation reaction 3h under room temperature, add ICAM-150 μ g monoclonal antibody, whirlpool Revolve and continue incubation reaction 3h under room temperature after 1min is uniformly dispersed.This reaction by N, N '-two succinimidyl carbonate connect On pharmaceutical carrier, the amino of Polyethylene Glycol and the amino in ICAM-1 monoclonal antibody, be successfully grafted onto pharmaceutical carrier by antibody On, lyophilization i.e. obtains the simvastatin nano structured lipid carrier that ICAM-1 monoclonal antibody is modified.
Use the size of nanoparticle prepared by dynamic light scattering determination.Sample is diluted to deionized water 0.1mg/ml, measures pungent cutting down before and after ICAM-1 monoclonal antibody is modified with Malvern Zetasizer Nano-S90 nano-particle size analysis instrument The particle diameter of statin nano structured lipid carrier is respectively 302.5 ± 12.5nm, 311.5 ± 12.7nm.
Acid adjustment centrifuging is used to coordinate envelop rate and the load of Syrups by HPLC simvastatin nano structured lipid carrier Dose.Stand to add to the simvastatin nanoparticle dispersion liquid without the modification of ICAM-1 monoclonal antibody of room temperature after preparation and fit The hydrochloric acid of amount 0.1M, the PH to system is down to 1.2 to promote nanoparticle flocculate, and 20000rpm is centrifuged 30min, HPLC mensuration supernatant Liquid drug concentration, calculates envelop rate and the drug loading of simvastatin nano structured lipid carrier as follows.
Envelop rate=(drug quality in lipid nanoparticle/total dosage) × 100%
Drug loading=(drug quality in lipid nanoparticle/drug-carrying nanometer particle gross mass) × 100%
After measured, envelop rate and the drug loading of the simvastatin nano structured lipid carrier modified without ICAM-1 monoclonal antibody divide It is not 87.93% and 4.42%.
The preparation of the simvastatin nano structured lipid carrier that embodiment 11 ICAM-1 monoclonal antibody is modified
Solvent diffusion method prepares simvastatin nano structured lipid carrier, concrete grammar: precision weigh 5mg simvastatin, 74mg glyceryl monostearate, 20mg medium chain length fatty acid triglyceride, the Polyethylene Glycol stearic acid of the mono amino terminal of 1mg synthesis Ester, altogether 100mg, add 1ml ethanol and dissolve as organic facies, water-bath 60 DEG C heating, make material melt completely.Syringe is drawn The material that 0.45ml melts, ethanol dilution to 0.5ml, with deionized water as disperse medium, will have under 400rpm mixing speed In the deionized water of the 4.5ml that machine is rapidly injected 60 DEG C of heating in water bath mutually successively, take out after injecting complete stirring 30s, the coldest But to room temperature, the simvastatin nano structured lipid carrier of lipid concentration 9mg/ml is prepared.Add N, N '-two succinum of 0.1mg Imide carbonic ester, vortex 1min be uniformly dispersed after incubation reaction 3h under room temperature, add 50 μ g ICAM-1 monoclonal antibodies, Vortex 1min continues incubation reaction 3h under room temperature after being uniformly dispersed.This reaction by N, N '-two succinimidyl carbonate even Connect the amino of Polyethylene Glycol and the amino in ICAM-1 monoclonal antibody on pharmaceutical carrier, antibody is successfully grafted onto pharmaceutical carrier On, lyophilization i.e. obtains the simvastatin nano structured lipid carrier that ICAM-1 monoclonal antibody is modified.
Use the size of nanoparticle prepared by dynamic light scattering determination.Sample is diluted to deionized water 0.1mg/ml, measures pungent cutting down before and after ICAM-1 monoclonal antibody is modified with Malvern Zetasizer Nano-S90 nano-particle size analysis instrument The particle diameter of statin nano structured lipid carrier is respectively 322.1 ± 17.7nm, 330.6 ± 20.2nm.
Acid adjustment centrifuging is used to coordinate envelop rate and the load of Syrups by HPLC simvastatin nano structured lipid carrier Dose.Stand to add to the simvastatin nanoparticle dispersion liquid without the modification of ICAM-1 monoclonal antibody of room temperature after preparation and fit The hydrochloric acid of amount 0.1M, the PH to system is down to 1.2 to promote nanoparticle flocculate, and 20000rpm is centrifuged 30min, HPLC mensuration supernatant Liquid drug concentration, calculates envelop rate and the drug loading of simvastatin nano structured lipid carrier as follows.
Envelop rate=(drug quality in lipid nanoparticle/total dosage) × 100%
Drug loading=(drug quality in lipid nanoparticle/drug-carrying nanometer particle gross mass) × 100%
After measured, envelop rate and the drug loading of the simvastatin nano structured lipid carrier modified without ICAM-1 monoclonal antibody divide It is not 92.19% and 4.63%.
The preparation of the simvastatin nano structured lipid carrier that embodiment 12 ICAM-1 monoclonal antibody is modified
Solvent diffusion method prepares simvastatin nano structured lipid carrier, concrete grammar: precision weigh 5mg simvastatin, 64mg glyceryl monostearate, 30mg medium chain length fatty acid triglyceride, the Polyethylene Glycol stearic acid of the mono amino terminal of 1mg synthesis Ester, altogether 100mg, add 1ml ethanol and dissolve as organic facies, water-bath 60 DEG C heating, make material melt completely.Syringe is drawn The material that 0.45ml melts, ethanol dilution to 0.5ml, with deionized water as disperse medium, will have under 400rpm mixing speed In the deionized water of the 4.5ml that machine is rapidly injected 60 DEG C of heating in water bath mutually successively, take out after injecting complete stirring 30s, the coldest But to room temperature, the simvastatin nano structured lipid carrier of lipid concentration 9mg/ml is prepared.Add N, N '-two succinum of 0.1mg Imide carbonic ester, vortex 1min be uniformly dispersed after incubation reaction 3h under room temperature, add 50 μ g ICAM-1 monoclonal antibodies, Vortex 1min continues incubation reaction 3h under room temperature after being uniformly dispersed.This reaction by N, N '-two succinimidyl carbonate even Connect the amino of Polyethylene Glycol and the amino in ICAM-1 monoclonal antibody on pharmaceutical carrier, antibody is successfully grafted onto pharmaceutical carrier On, lyophilization i.e. obtains the simvastatin nano structured lipid carrier that ICAM-1 monoclonal antibody is modified.
Use the size of nanoparticle prepared by dynamic light scattering determination.Sample is diluted to deionized water 0.1mg/ml, measures pungent cutting down before and after ICAM-1 monoclonal antibody is modified with Malvern Zetasizer Nano-S90 nano-particle size analysis instrument The particle diameter of statin nano structured lipid carrier is respectively 345.1 ± 12.7nm, 355.6 ± 23.7nm.
Acid adjustment centrifuging is used to coordinate envelop rate and the load of Syrups by HPLC simvastatin nano structured lipid carrier Dose.Stand to add to the simvastatin nanoparticle dispersion liquid without the modification of ICAM-1 monoclonal antibody of room temperature after preparation and fit The hydrochloric acid of amount 0.1M, the PH to system is down to 1.2 to promote nanoparticle flocculate, and 20000rpm is centrifuged 30min, HPLC mensuration supernatant Liquid drug concentration, calculates envelop rate and the drug loading of simvastatin nano structured lipid carrier as follows.
Envelop rate=(drug quality in lipid nanoparticle/total dosage) × 100%
Drug loading=(drug quality in lipid nanoparticle/drug-carrying nanometer particle gross mass) × 100%
After measured, envelop rate and the drug loading of the simvastatin nano structured lipid carrier modified without ICAM-1 monoclonal antibody divide It is not 91.18% and 4.58%.、
The preparation of the simvastatin nano structured lipid carrier that embodiment 13 ICAM-1 monoclonal antibody is modified
Solvent diffusion method prepares simvastatin nano structured lipid carrier, concrete grammar: precision weigh 5mg simvastatin, 74mg glyceryl monostearate, 10mg oleic acid, the poly-second of the mono amino terminal of 10mg polyethylene glycol mono stearate, 1mg synthesis Glycol stearate, altogether 100mg, add 1ml ethanol and dissolve as organic facies, water-bath 60 DEG C heating, make material melt completely.Note Emitter draws the material that 0.45ml melts, and ethanol dilution to 0.5ml, with deionized water as disperse medium, in 400rpm stirring speed In the deionized water of the 4.5ml that under degree, organic facies is rapidly injected successively 60 DEG C of heating in water bath, take after injecting complete stirring 30s Go out, naturally cool to room temperature, prepare the simvastatin nano structured lipid carrier of lipid concentration 9mg/ml.Add the N of 0.1mg, N '-two succinimidyl carbonate, vortex 1min be uniformly dispersed after incubation reaction 3h under room temperature, add 50 μ g ICAM-1 mono- Clonal antibody, vortex 1min continues incubation reaction 3h under room temperature after being uniformly dispersed.This reaction is by N, N '-two succinimido Carbonic ester connects the amino of Polyethylene Glycol and the amino in ICAM-1 monoclonal antibody on pharmaceutical carrier, is successfully grafted onto by antibody On pharmaceutical carrier, lyophilization i.e. obtains the simvastatin nano structured lipid carrier that ICAM-1 monoclonal antibody is modified.
Use the size of nanoparticle prepared by dynamic light scattering determination.Sample is diluted to deionized water 0.1mg/ml, measures pungent cutting down before and after ICAM-1 monoclonal antibody is modified with Malvern Zetasizer Nano-S90 nano-particle size analysis instrument The particle diameter of statin nano structured lipid carrier is respectively 295.5 ± 13.7nm, 310.4 ± 14.8nm.
Acid adjustment centrifuging is used to coordinate envelop rate and the load of Syrups by HPLC simvastatin nano structured lipid carrier Dose.Stand to add to the simvastatin nanoparticle dispersion liquid without the modification of ICAM-1 monoclonal antibody of room temperature after preparation and fit The hydrochloric acid of amount 0.1M, the PH to system is down to 1.2 to promote nanoparticle flocculate, and 20000rpm is centrifuged 30min, HPLC mensuration supernatant Liquid drug concentration, calculates envelop rate and the drug loading of simvastatin nano structured lipid carrier as follows.
Envelop rate=(drug quality in lipid nanoparticle/total dosage) × 100%
Drug loading=(drug quality in lipid nanoparticle/drug-carrying nanometer particle gross mass) × 100%
After measured, envelop rate and the drug loading of the simvastatin nano structured lipid carrier modified without ICAM-1 monoclonal antibody divide It is not 82.25% and 4.15%.
The preparation of the simvastatin nano structured lipid carrier that embodiment 14 ICAM-1 monoclonal antibody is modified
Solvent diffusion method prepares simvastatin nano structured lipid carrier, concrete grammar: precision weigh 5mg simvastatin, 64mg glyceryl monostearate, 20mg oleic acid, the poly-second of the mono amino terminal of 10mg polyethylene glycol mono stearate, 1mg synthesis Glycol stearate, altogether 100mg, add 1ml ethanol and dissolve as organic facies, water-bath 60 DEG C heating, make material melt completely.Note Emitter draws the material that 0.45ml melts, and ethanol dilution to 0.5ml, with deionized water as disperse medium, in 400rpm stirring speed In the deionized water of the 4.5ml that under degree, organic facies is rapidly injected successively 60 DEG C of heating in water bath, take after injecting complete stirring 30s Go out, naturally cool to room temperature, prepare the simvastatin nano structured lipid carrier of lipid concentration 9mg/ml.Add the N of 0.1mg, N '-two succinimidyl carbonate, vortex 1min be uniformly dispersed after incubation reaction 3h under room temperature, add 50 μ g ICAM-1 mono- Clonal antibody, vortex 1min continues incubation reaction 3h under room temperature after being uniformly dispersed.This reaction is by N, N '-two succinimido Carbonic ester connects the amino of Polyethylene Glycol and the amino in ICAM-1 monoclonal antibody on pharmaceutical carrier, is successfully grafted onto by antibody On pharmaceutical carrier, lyophilization i.e. obtains the simvastatin nano structured lipid carrier that ICAM-1 monoclonal antibody is modified.
Use the size of nanoparticle prepared by dynamic light scattering determination.Sample is diluted to deionized water 0.1mg/ml, measures pungent cutting down before and after ICAM-1 monoclonal antibody is modified with Malvern Zetasizer Nano-S90 nano-particle size analysis instrument The particle diameter of statin nano structured lipid carrier is respectively 320.7 ± 20.1nm, 325.0 ± 19.2nm.
Acid adjustment centrifuging is used to coordinate envelop rate and the load of Syrups by HPLC simvastatin nano structured lipid carrier Dose.Stand to add to the simvastatin nanoparticle dispersion liquid without the modification of ICAM-1 monoclonal antibody of room temperature after preparation and fit The hydrochloric acid of amount 0.1M, the PH to system is down to 1.2 to promote nanoparticle flocculate, and 20000rpm is centrifuged 30min, HPLC mensuration supernatant Liquid drug concentration, calculates envelop rate and the drug loading of simvastatin nano structured lipid carrier as follows.
Envelop rate=(drug quality in lipid nanoparticle/total dosage) × 100%
Drug loading=(drug quality in lipid nanoparticle/drug-carrying nanometer particle gross mass) × 100%
After measured, envelop rate and the drug loading of the simvastatin nano structured lipid carrier modified without ICAM-1 monoclonal antibody divide It is not 86.55% and 4.36%.
The preparation of the simvastatin nano structured lipid carrier that embodiment 15 ICAM-1 monoclonal antibody is modified
Solvent diffusion method prepares simvastatin nano structured lipid carrier, concrete grammar: precision weigh 5mg simvastatin, 54mg glyceryl monostearate, 30mg oleic acid, the poly-second of the mono amino terminal of 10mg polyethylene glycol mono stearate, 1mg synthesis Glycol stearate, altogether 100mg, add 1ml ethanol and dissolve as organic facies, water-bath 60 DEG C heating, make material melt completely.Note Emitter draws the material that 0.45ml melts, and ethanol dilution to 0.5ml, with deionized water as disperse medium, in 400rpm stirring speed In the deionized water of the 4.5ml that under degree, organic facies is rapidly injected successively 60 DEG C of heating in water bath, take after injecting complete stirring 30s Go out, naturally cool to room temperature, prepare the simvastatin nano structured lipid carrier of lipid concentration 9mg/ml.Add the N of 0.1mg, N '-two succinimidyl carbonate, vortex 1min be uniformly dispersed after incubation reaction 3h under room temperature, add 50 μ g ICAM-1 mono- Clonal antibody, vortex 1min continues incubation reaction 3h under room temperature after being uniformly dispersed.This reaction is by N, N '-two succinimido Carbonic ester connects the amino of Polyethylene Glycol and the amino in ICAM-1 monoclonal antibody on pharmaceutical carrier, is successfully grafted onto by antibody On pharmaceutical carrier, lyophilization i.e. obtains the simvastatin nano structured lipid carrier that ICAM-1 monoclonal antibody is modified.
Use the size of nanoparticle prepared by dynamic light scattering determination.Sample is diluted to deionized water 0.1mg/ml, measures pungent cutting down before and after ICAM-1 monoclonal antibody is modified with Malvern Zetasizer Nano-S90 nano-particle size analysis instrument The particle diameter of statin nano structured lipid carrier is respectively 340.2 ± 23.3nm, 345.9 ± 15.1nm.
Acid adjustment centrifuging is used to coordinate envelop rate and the load of Syrups by HPLC simvastatin nano structured lipid carrier Dose.Stand to add to the simvastatin nanoparticle dispersion liquid without the modification of ICAM-1 monoclonal antibody of room temperature after preparation and fit The hydrochloric acid of amount 0.1M, the PH to system is down to 1.2 to promote nanoparticle flocculate, and 20000rpm is centrifuged 30min, HPLC mensuration supernatant Liquid drug concentration, calculates envelop rate and the drug loading of simvastatin nano structured lipid carrier as follows.
Envelop rate=(drug quality in lipid nanoparticle/total dosage) × 100%
Drug loading=(drug quality in lipid nanoparticle/drug-carrying nanometer particle gross mass) × 100%
After measured, envelop rate and the drug loading of the simvastatin nano structured lipid carrier modified without ICAM-1 monoclonal antibody divide It is not 85.22% and 4.29%.
The preparation of the simvastatin nano structured lipid carrier that embodiment 16 ICAM-1 monoclonal antibody is modified
Solvent diffusion method prepares simvastatin nano structured lipid carrier, concrete grammar: precision weigh 5mg simvastatin, 74mg glyceryl monostearate, the monoamine of 10mg medium chain length fatty acid triglyceride, 10mg polyethylene glycol mono stearate, 1mg synthesis The polyglycol distearate of base terminal, altogether 100mg, add 1ml ethanol and dissolve as organic facies, water-bath 60 DEG C heating, make material Material melts completely.The material that syringe absorption 0.45ml melts, ethanol dilution to 0.5ml, with deionized water as disperse medium, In the deionized water of the 4.5ml that under 400rpm mixing speed, organic facies is rapidly injected successively 60 DEG C of heating in water bath, inject complete Take out after stirring 30s, naturally cool to room temperature, prepare the simvastatin nano structured lipid carrier of lipid concentration 9mg/ml.Add Enter the N of 0.1mg, N '-two succinimidyl carbonate, vortex 1min be uniformly dispersed after incubation reaction 3h under room temperature, add 50 μ G ICAM-1 monoclonal antibody, vortex 1min continues incubation reaction 3h under room temperature after being uniformly dispersed.This reaction is by N, N '-two amber Amber imide carbonic ester connects the amino of Polyethylene Glycol and the amino in ICAM-1 monoclonal antibody on pharmaceutical carrier, by antibody Successfully being grafted onto on pharmaceutical carrier, lyophilization i.e. obtains the simvastatin nano structured lipid carrier that ICAM-1 monoclonal antibody is modified.
Use the size of nanoparticle prepared by dynamic light scattering determination.Sample is diluted to deionized water 0.1mg/ml, measures pungent cutting down before and after ICAM-1 monoclonal antibody is modified with Malvern Zetasizer Nano-S90 nano-particle size analysis instrument The particle diameter of statin nano structured lipid carrier is respectively 317.5 ± 15.8nm, 323.9 ± 14.2nm.
Acid adjustment centrifuging is used to coordinate envelop rate and the load of Syrups by HPLC simvastatin nano structured lipid carrier Dose.Stand to add to the simvastatin nanoparticle dispersion liquid without the modification of ICAM-1 monoclonal antibody of room temperature after preparation and fit The hydrochloric acid of amount 0.1M, the PH to system is down to 1.2 to promote nanoparticle flocculate, and 20000rpm is centrifuged 30min, HPLC mensuration supernatant Liquid drug concentration, calculates envelop rate and the drug loading of simvastatin nano structured lipid carrier as follows.
Envelop rate=(drug quality in lipid nanoparticle/total dosage) × 100%
Drug loading=(drug quality in lipid nanoparticle/drug-carrying nanometer particle gross mass) × 100%
After measured, envelop rate and the drug loading of the simvastatin nano structured lipid carrier modified without ICAM-1 monoclonal antibody divide It is not 86.72% and 4.36%.
The preparation of the simvastatin nano structured lipid carrier that embodiment 17 ICAM-1 monoclonal antibody is modified
Solvent diffusion method prepares simvastatin nano structured lipid carrier, concrete grammar: precision weigh 5mg simvastatin, 64mg glyceryl monostearate, the monoamine of 20mg medium chain length fatty acid triglyceride, 10mg polyethylene glycol mono stearate, 1mg synthesis The polyglycol distearate of base terminal, altogether 100mg, add 1ml ethanol and dissolve as organic facies, water-bath 60 DEG C heating, make material Material melts completely.The material that syringe absorption 0.45ml melts, ethanol dilution to 0.5ml, with deionized water as disperse medium, In the deionized water of the 4.5ml that under 400rpm mixing speed, organic facies is rapidly injected successively 60 DEG C of heating in water bath, inject complete Take out after stirring 30s, naturally cool to room temperature, prepare the simvastatin nano structured lipid carrier of lipid concentration 9mg/ml.Add Enter the N of 0.1mg, N '-two succinimidyl carbonate, vortex 1min be uniformly dispersed after incubation reaction 3h under room temperature, add 50 μ G ICAM-1 monoclonal antibody, vortex 1min continues incubation reaction 3h under room temperature after being uniformly dispersed.This reaction is by N, N '-two amber Amber imide carbonic ester connects the amino of Polyethylene Glycol and the amino in ICAM-1 monoclonal antibody on pharmaceutical carrier, by antibody Successfully being grafted onto on pharmaceutical carrier, lyophilization i.e. obtains the simvastatin nano structured lipid carrier that ICAM-1 monoclonal antibody is modified.
Use the size of nanoparticle prepared by dynamic light scattering determination.Sample is diluted to deionized water 0.1mg/ml, measures pungent cutting down before and after ICAM-1 monoclonal antibody is modified with Malvern Zetasizer Nano-S90 nano-particle size analysis instrument The particle diameter of statin nano structured lipid carrier is respectively 337.8 ± 25.8nm, 354.7 ± 18.2nm.
Acid adjustment centrifuging is used to coordinate envelop rate and the load of Syrups by HPLC simvastatin nano structured lipid carrier Dose.Stand to add to the simvastatin nanoparticle dispersion liquid without the modification of ICAM-1 monoclonal antibody of room temperature after preparation and fit The hydrochloric acid of amount 0.1M, the PH to system is down to 1.2 to promote nanoparticle flocculate, and 20000rpm is centrifuged 30min, HPLC mensuration supernatant Liquid drug concentration, calculates envelop rate and the drug loading of simvastatin nano structured lipid carrier as follows.
Envelop rate=(drug quality in lipid nanoparticle/total dosage) × 100%
Drug loading=(drug quality in lipid nanoparticle/drug-carrying nanometer particle gross mass) × 100%
After measured, envelop rate and the drug loading of the simvastatin nano structured lipid carrier modified without ICAM-1 monoclonal antibody divide It is not 91.04% and 4.43%.
Embodiment 18
The preparation of the simvastatin nano structured lipid carrier that 1.ICAM-1 monoclonal antibody is modified
Solvent diffusion method prepares simvastatin nano structured lipid carrier, concrete grammar: precision weigh 5mg simvastatin, 54mg glyceryl monostearate, the monoamine of 30mg medium chain length fatty acid triglyceride, 10mg polyethylene glycol mono stearate, 1mg synthesis The polyglycol distearate of base terminal, altogether 100mg, add 1ml ethanol and dissolve as organic facies, water-bath 60 DEG C heating, make material Material melts completely.The material that syringe absorption 0.45ml melts, ethanol dilution to 0.5ml, with deionized water as disperse medium, In the deionized water of the 4.5ml that under 400rpm mixing speed, organic facies is rapidly injected successively 60 DEG C of heating in water bath, inject complete Take out after stirring 30s, naturally cool to room temperature, prepare the simvastatin nano structured lipid carrier of lipid concentration 9mg/ml.Add Enter the N of 0.1mg, N '-two succinimidyl carbonate, vortex 1min be uniformly dispersed after incubation reaction 3h under room temperature, add 50 μ G ICAM-1 monoclonal antibody, vortex 1min continues incubation reaction 3h under room temperature after being uniformly dispersed.This reaction is by N, N '-two amber Amber imide carbonic ester connects the amino of Polyethylene Glycol and the amino in ICAM-1 monoclonal antibody on pharmaceutical carrier, by antibody Successfully being grafted onto on pharmaceutical carrier, lyophilization i.e. obtains the simvastatin nano structured lipid carrier that ICAM-1 monoclonal antibody is modified.
Use the size of nanoparticle prepared by dynamic light scattering determination.Sample is diluted to deionized water 0.1mg/ml, measures pungent cutting down before and after ICAM-1 monoclonal antibody is modified with Malvern Zetasizer Nano-S90 nano-particle size analysis instrument The particle diameter of statin nano structured lipid carrier is respectively 342.5 ± 23.6nm, 358.1 ± 22.7nm.
Acid adjustment centrifuging is used to coordinate envelop rate and the load of Syrups by HPLC simvastatin nano structured lipid carrier Dose.Stand to add to the simvastatin nanoparticle dispersion liquid without the modification of ICAM-1 monoclonal antibody of room temperature after preparation and fit The hydrochloric acid of amount 0.1M, the PH to system is down to 1.2 to promote nanoparticle flocculate, and 20000rpm is centrifuged 30min, HPLC mensuration supernatant Liquid drug concentration, calculates envelop rate and the drug loading of simvastatin nano structured lipid carrier as follows.
Envelop rate=(drug quality in lipid nanoparticle/total dosage) × 100%
Drug loading=(drug quality in lipid nanoparticle/drug-carrying nanometer particle gross mass) × 100%
After measured, envelop rate and the drug loading of the simvastatin nano structured lipid carrier modified without ICAM-1 monoclonal antibody divide It is not 87.02% and 4.38%.
2. the morphologic observation of simvastatin nano structured lipid carrier
In treating excess syndrome example 17, the lipid nanoparticle before lyophilization is diluted to 100 μ g/ml, nanoparticle dispersion liquid after diluting Dropping on copper mesh, natural drying, with uranium acetate solution negative staining 1min of 1%, JEM-1200EX type transmission electron microscope Observe its form.Simvastatin nano structured lipid carrier form before and after ICAM-1 monoclonal antibody is modified is as it is shown in figure 1, in class ball Shape；Nanoparticle size before and after monoclonal antibody is modified is close.
3. the vitro drug release of simvastatin nano structured lipid carrier is investigated
In treating excess syndrome example 17, the simvastatin nano structured lipid carrier dispersion liquid before lyophilization dilutes 9 times to dispersion liquid Drug concentration is 50 μ g/ml.Accurately weighed simvastatin 10mg, certain ethanol is diluted to the PBS solution of PH7.4 after dissolving 50μg/ml.Use Bag filter method, take respectively after simvastatin standard solution and dilution through antibody modification or unmodified Simvastatin nano structured lipid carrier solution 1ml, in bag filter, is placed in the PBS release pipe containing 15ml PH7.4,37 DEG C, 60rpm isothermal vibration.Set point in time sampling 1ml, and supply the fresh PBS solution of 1ml.Syrups by HPLC discharges Simvastatin concentration in medium, calculates the cumulative release amount of simvastatin.Simvastatin nano structured lipid carrier after measured Release behavior is as in figure 2 it is shown, compared with simple free drug, drug-carrying nanometer particle has slow releasing function to medicine；ICAM-1 monoclonal antibody Simvastatin nano structured lipid carrier release in vitro behavior before and after modification is similar.
4. the structure of acute lung injury animal model and the lung of ICAM-1 monoclonal antibody simvastatin nano structured lipid carrier Portion's targeting research
Animal model is built with Balb/c mice.Carry out anaesthetizing (450mg/kg) with 5% chloral hydrate by mice, exposed Tracheal instillation method, slowly instils and is dissolved in the lipopolysaccharide (2 μ g/g, bodyweight) of 50 μ l normal saline, is administered and carries out medicine after 6h Effect is evaluated.
For pulmonary's targeting of ICAM-1 monoclonal antibody simvastatin nano structured lipid carrier in evaluation example 17, with Dir Wrapped in loading lipid nanoparticle as fluorescent probe.Balb/c mice is carried out following packet: A group: normal mice+ICAM-1 Monoclonal antibody simvastatin nano structured lipid carrier group；B group: instillation normal saline Mus+ICAM-1 monoclonal antibody simvastatin nanostructured Lipid carrier group；C group: model mouse+ICAM-1 monoclonal antibody simvastatin nano structured lipid carrier group；D group: model mouse+pungent cuts down him Spit of fland nano structured lipid carrier group.After modeling 6h, tail vein Bolos intravenous administration.After being administered, 5h, 24h put to death mice, live body Imager is observed and takes pictures.It is observed that ICAM-1 monoclonal antibody modify before and after the distribution in vivo of simvastatin nano structured lipid carrier Characteristic is as it is shown on figure 3, ICAM-1 monoclonal antibody simvastatin nano structured lipid carrier is just significantly stronger than in the distribution of model mice pulmonary Often mice and expose the mice of tracheal instillation normal saline to the open air, illustrate after LPS stimulation, model group mouse lung vascular endothelial cell A large amount have expressed ICAM-1 adhesion molecule, and ICAM-1 monoclonal antibody is successfully grafted onto on nano-carrier, by antibody receptor specificity In conjunction with, ICAM-1 monoclonal antibody and nano structured lipid carrier is distributed in the lung of model mice in a large number；ICAM-1 monoclonal antibody is repaiied simultaneously The nano structured lipid carrier of decorations is distributed more in the lung of model mouse compared with the nano structured lipid carrier modified without ICAM-1 monoclonal antibody Many, embody pulmonary's targeting characteristic of ICAM-1 monoclonal antibody simvastatin nano structured lipid carrier.
The protective effect to acute lung injury of the 5.HE staining evaluation ICAM-1 monoclonal antibody simvastatin nano structured lipid carrier For the ICAM-1 monoclonal antibody simvastatin nano structured lipid carrier protective effect to acute lung injury in evaluation example 17, incite somebody to action Balb/c mice carries out following packet, A group: matched group；B group: model group；C group: model+free simvastatin administration group；D group: Model+simvastatin nano structured lipid carrier administration group；E group: model+ICAM-1 monoclonal antibody simvastatin nanostructured lipid carries Body administration group.After mice modeling 6h, A group, B group tail vein injection 0.2ml normal saline；C group tail vein injection 200 μ g/ml's Free simvastatin drug solution；D group tail vein injection simvastatin nano structured lipid carrier；E group tail vein injection ICAM- 1 monoclonal antibody simvastatin nano structured lipid carrier (dosage of each administration group simvastatin is 2mg/kg).It is administered 24h Rear execution mice, takes out lung organ and fixes 48h, HE dyeing with 4% neutral formalin, and light Microscopic observation respectively organizes lung pathologies section Feature, evaluates the improvement situation of the chmice acute injury of lung that LPS is induced by pharmaceutical preparation.Result as shown in Figure 4, free pungent cuts down him Spit of fland drug solution group, simvastatin nano structured lipid carrier group and ICAM-1 monoclonal antibody simvastatin nano structured lipid carrier Lung tissue's section pathologic condition of group mice all obtains a certain degree of improvement compared with model group.The pathological section for the treatment of group mice Photo is for model group: in alveolar, the inflammatory cell of infiltration significantly reduces, and alveolar structure is visible, and wherein ICAM-1 is pungent cuts down It is the most notable that statin nano structured lipid carrier experimental group improves situation.Analysis result shows, ICAM-1 simvastatin nano junction Animal pattern acute lung injury disease in this problem can be played a protective role by structure lipid carrier.

Claims (6)

1. an ICAM-1 monoclonal antibody modifies simvastatin nano structured lipid carrier, it is characterised in that described lipid carrier is by pungent Cut down statin, matrix material, polyethylene glycol mono stearate (MW 2000), the polyglycol distearate of mono amino terminal, N, N '-two succinimidyl carbonate and ICAM-1 monoclonal antibody composition, mass percent is that 4.98%-14.95% is pungent Cut down statin, the matrix material of 83.72%-93.68%, the polyethylene glycol mono stearate (MW 2000) of 0%-9.97%, The polyglycol distearate of mono amino terminal of 1.00%, the N of 0.22%, N '-two succinimidyl carbonate and The ICAM-1 monoclonal antibody of 0.11%.
A kind of ICAM-1 monoclonal antibody the most according to claim 1 modifies simvastatin nano structured lipid carrier, and its feature exists In, described matrix material selects the one or two in glyceryl monostearate, stearic acid, oleic acid, medium chain length fatty acid triglyceride Kind.
3. a kind of ICAM-1 monoclonal antibody described in claim 1 modifies the preparation method of simvastatin nano structured lipid carrier, its It is characterised by, is realized by following steps:
(1) polyglycol distearate of mono amino terminal is synthesized
Weigh 1-(3-dimethylamino-propyl)-3-ethyl-carbodiimide hydrochloride, N-hydroxy-succinamide, stearic acid are dissolved in nothing Water dimethyl sulfoxide, reacts 30min under the conditions of 400rpm 60 DEG C, be slowly added dropwise by reactant liquor to anhydrous dimethyl sulphoxide dissolving Double N-terminal polyglycol solutions in, under room temperature 400rpm stirring reaction 24h, reaction terminate after, reactant liquor is transferred to In analysis bag, pure water dialysis 48h, remove aqueous by-product, lyophilization i.e. obtains the Polyethylene Glycol stearic acid of mono amino terminal Ester；
(2) preparation of the simvastatin nano structured lipid carrier that ICAM-1 monoclonal antibody is modified
Solvent diffusion method prepares simvastatin nano structured lipid carrier, concrete grammar: weighs simvastatin, matrix material, gather Ethylene glycol stearate and the polyethylene glycol mono stearate of N-terminal, add ethanol and dissolve as organic facies, water-bath 60 DEG C heating, make material melt completely, syringe draw melt material, ethanol dilution, with deionized water as disperse medium, Under 400rpm mixing speed, organic facies is rapidly injected successively 60 DEG C of heating in water bath deionized water in, inject complete stirring Take out after 30s, naturally cool to room temperature, prepare simvastatin nano structured lipid carrier, add N, N '-two butanimide Base carbonic ester, vortex 1min be uniformly dispersed after incubation reaction 3h under room temperature, add ICAM-1 monoclonal antibody, vortex 1min disperses Continuing incubation reaction 3h after uniformly under room temperature, lyophilization i.e. obtains ICAM-1 monoclonal antibody and modifies simvastatin nanostructured lipid load Body.
A kind of ICAM-1 monoclonal antibody the most according to claim 3 modifies the preparation side of simvastatin nano structured lipid carrier Method, it is characterised in that step (1) bag filter is MWCO:7kDa.
A kind of ICAM-1 monoclonal antibody the most according to claim 3 modifies the preparation side of simvastatin nano structured lipid carrier Method, it is characterised in that step (2) polyglycol distearate selects MW 2000.
A kind of ICAM-1 monoclonal antibody the most according to claim 1 is modified simvastatin nano structured lipid carrier and is being prepared targeting Application in ALI pulmonary vascular endothelial cell medicine.
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CN112220932B (en) * 2020-10-21 2022-12-13 浙江省立同德医院 VCAM-1 monoclonal antibody tanshinone IIA nanostructured lipid carrier, preparation method and application

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