CN-118892454-B - Magnetic drug-loaded liposome for nerve repair and preparation method thereof

Abstract

The invention discloses a magnetic drug-loaded liposome for nerve repair and a preparation method thereof, wherein the magnetic drug-loaded liposome prepared by the method comprises magnetic nano particles and drugs, wherein the mass fractions of egg yolk lecithin, cholesterol, DSPE-MPEG 2000, BFe 3 O 4 and drugs in 1mL liposome solution are respectively 0.5% -1.5%, 0.15% -0.35%, 0.1% -0.2% and 0.1% -0.2%. The medicine is one or more of taxifolin, resveratrol or tea polyphenol in natural antioxidant medicine. The multifunctional magnetic drug-loaded liposome provided by the invention can be used for repairing nerve cells after injury in combination with loading of a magnetic field.

Inventors

• LIU MEILI
• SUN XUEMEI
• LI PING
• CHENG YUJING
• HUANG ZIYANG
• Bai Ruoyan

Assignees

• 北京航空航天大学

Dates

Publication Date

20260505

Application Date

20240710

Claims (1)

1. 1. A method for preparing a magnetic drug-loaded liposome for nerve repair, which is characterized by comprising the following steps: the designed target components are egg yolk lecithin, cholesterol, DSPE-MPEG2000, in 1mL liposome solution, The mass fractions of the medicines are respectively 0.5% -1.5%, 0.15% -0.35%, 0.1% -0.2% and 0.1% -0.2%; step one, adopting a coprecipitation method to prepare Magnetic nanoparticles; Adding into deionized water And After magnetically stirring for 0.5h at the water bath temperature of 70-85 ℃, ammonium hydroxide is injected, stirring is continued for 3-4 h, the product is taken out, washed by deionized water twice and ethanol twice, and dried for 18-24 h in a freeze dryer at the temperature of-30-50 ℃ to obtain Magnetic nanoparticles; the dosage is 1 g-3 g of deionized water required by 100mL 2 G-4 g 10-20 ML of ammonium hydroxide; the mass percentage concentration of the ammonium hydroxide is 25%; The speed regulation of the magnetic stirrer is 400 rpm-1000 rpm; Modification of magnetic nano particles; Bovine serum albumin is added to In the magnetic nanoparticle solution, stirring vigorously at 0deg.C for 10h to obtain A solution; The dosage is 10mg Adding 30-80 mg of bovine serum albumin into the magnetic nano particles; The speed of the magnetic stirrer is 800 rpm-1200 rpm; step three, preparing a lipid film by decompression rotary evaporation; dissolving egg yolk lecithin, cholesterol, DSPE-PEG2000 and a drug in an ethanol solution, carrying out ultrasonic vortex mixing for 2min, transferring the mixed solution into a round-bottom flask of a rotary evaporator, carrying out reduced pressure rotary evaporation at 50 ℃ and 50-100 rpm of rotation speed to remove ethanol, and finally forming a uniform lipid film at the bottom of the round-bottom flask; the medicament is a natural antioxidant for nerve repair, in particular to one or more than one of taxifolin or tea polyphenol; the dosage of the medicine comprises 30-90 mg of egg yolk lecithin, 5-15 mg of cholesterol, 5-15 mg of DSPE-PEG2000 and 3-9 mg of medicine required in 50mL of ethanol; the ultrasonic frequency is 60kHz; DSPE-PEG2000 refers to distearoyl phosphatidylethanolamine-polyethylene glycol 2000; Step four, preparing liposome after hydration; adding into rotary evaporation bottle The solution is hydrated, ultrasonic until the lipid film at the bottom of the rotary evaporation bottle falls off completely, then the solution is placed in a water bath at the temperature of 30-60 ℃ and stirred continuously, a rubber head suction pipe is used for blowing the rotary evaporation bottle to collect lipid suspension, the collected suspension is transferred into a sample bottle, ultrasonic treatment is carried out for 3-10 min by a probe of a cell breaker at the temperature of 0 ℃ of ice bath, namely, ultrasonic treatment is carried out for 3s, 2s is stopped, and finally the product is filtered to obtain a transparent and uniform magnetic drug carrying liposome solution; the ultrasonic frequency is 60kHz; the speed regulation of the magnetic stirrer is 350 rpm-600 rpm; The power of the cell disruption instrument is 150W-300W; the prepared magnetic drug-loaded liposome has a phospholipid bilayer structure, and PEG is modified on the surface to wrap magnetic nano particles and drugs.

Description

Magnetic drug-loaded liposome for nerve repair and preparation method thereof Technical Field The invention relates to a preparation method of liposome, in particular to a magnetic drug-loaded liposome for nerve repair and a preparation method thereof. Background Ischemic stroke is a clinical syndrome of a series of neurological deficit symptoms due to local or global ischemia and hypoxia necrosis of brain tissues caused by stenosis or occlusion of cerebral blood supply arteries (such as carotid artery and vertebral artery) and cerebral blood supply insufficiency. The incidence rate and the death rate of cerebral arterial thrombosis are high, and the cerebral arterial thrombosis is one of the most serious public health problems accepted in the world. Ischemia initiates an ischemic cascade involving a range of biochemical reactions including energy failure, ion imbalance and excitotoxicity, oxidative stress, cell death (apoptosis or necrosis), activation of the complement system, inflammation and initiation of immune responses, leading to irreversible damage. The clinical treatment for ischemic stroke is thrombolytic surgery using tissue-type plasminogen activator to achieve vascular recanalization. However, the time window for thrombolytic therapy is relatively narrow, so that only a few patients can receive thrombolytic therapy in time and benefit from it. The neurons of the central nervous system have the characteristics of non-regeneration and non-division proliferation, and how to repair nerves after nerve injury and block apoptosis of the neurons clearly becomes a research hot spot in the neuroacademic community. Liposomes are in the form of closed phospholipid bilayer spheres, with dimensions on the nano-scale, and are widely used as carriers for encapsulation of genes, drugs and imaging agents for the diagnosis and treatment of central nervous system diseases due to their excellent biocompatibility. The liposome has a unique oil-water amphiphilic structure and can wrap hydrophilic and hydrophobic compounds. The core space is hydrophilic, can wrap hydrophilic drugs and nucleic acid, is used as a carrier to convey the wrapping material to a target disease area, and can also slowly release the drugs to a certain extent. The phospholipid bilayer is hydrophobic and can encapsulate lipophilic compound. However, nanoliposomes have a problem of being easily phagocytosed or adsorbed by other substances in blood in practical application, and in order to solve the problem, researchers modify various polymers, targeting molecules or aptamers on the surface of the nanoliposomes, and the efficiency of the nanoliposomes in tissue transfer is further improved. Douglas fir element (taxifolin), also known as dihydroquercetin (dihydroquercetin), is a bioflavonoid substance extracted from the root of Larix Gmelini, a natural powerful antioxidant. Studies show that the douglas fir extract has outstanding medicinal values in the aspects of antioxidation, anti-inflammation, liver protection, anti-tumor, blood lipid regulation, anti-angiogenesis, anti-Alzheimer disease, antibiosis and the like. In addition, it can prevent liver injury due to carbon tetrachloride and inhibit erythrocyte lysis due to oxidation. As an excellent anti-inflammatory substance, the expression of interleukin-6 and interleukin-8 in tissues can be effectively reduced. Tea polyphenol, a compound widely existing in tea leaves, is considered to have various physiological activities such as antioxidation, anti-inflammatory, anticancer, blood pressure reduction, blood sugar reduction, aging resistance and the like, is helpful for preventing various diseases, and is beneficial to cardiovascular system, immune system and metabolism. The antioxidant activity is 2.6 times that of Butylated Hydroxyanisole (BHA) and 3.6 times that of vitamin E. Resveratrol mainly derived from peanut, grape (red wine), rhizoma Polygoni Cuspidati, mori fructus, etc., is a natural polyphenol compound and also a natural antioxidant. Research shows that resveratrol has physiological activities of resisting oxidation, resisting inflammation, resisting cancer, reducing blood lipid, reducing blood sugar, promoting cardiovascular health, etc. In the aspect of neuroprotection, the nerve protecting agent can remove free radicals in a human body, reduce damage of oxidative stress to nerve cells, promote growth and regeneration of the nerve cells, and play a role in neuroprotection by regulating various signal paths, such as signal transduction of nerve growth factors and the like. Disclosure of Invention The invention aims at providing a magnetic drug-loaded liposome with multiple functions of resisting oxidization, resisting apoptosis, promoting nerve regeneration and the like. The liposome has a phospholipid bilayer structure, and is coated with PEG (polyethylene glycol) and coated with magnetic nanoparticles and medicines, and on the other hand, the liposome combines the excellent magnet