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Target preparation consisting of liposome and nucleic acid coating contrast agent
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A technology of contrast agent and liposome, applied in the field of biomedicine
Inactive Publication Date: 2011-01-19
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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[0004] There is no targeting agent consisting of liposomes and nucleic acids encapsulating contrast agents for both diagnosis and gene therapy
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Embodiment 1
[0033] 1 Preparation of magnetically targeted liposomes
[0034] 1.1 Accurately weigh 20 mg of dioleoylphosphatidylethanolamine (DOPE, Fluka Company), 3β-[N-(N', N'-dimethylaminoethyl)] carbamoyl-cholesterol (DC-Chol, Sigma Company) 20mg, cholesterol (Chol, Japan Wako Pure Chemical Industry Co., Ltd.) 5mg and α-tocopherol (Shanghai No. 2 Pharmaceutical Factory) 1mg were placed in a 250ml round bottom flask, and 8ml of chloroform was added to dissolve it completely.
[0035] 1.2 Dilute ferric oxide tartaric acid solution with water for injection to 5 mg / ml in a cell pulverizer (JY92-IIN, Ningbo Xinzhi Biotechnology Co., Ltd.) and ultrasonically (400w, 5s each time, 40 times) to disperse Evenly, take 1.5ml and put it into the above round-bottomed flask, and ultrasonically (500w) for 30min in a water bath ultrasonic cleaner (SK5200LH, Shanghai Kedao Ultrasonic Instrument Co., Ltd.) to form a stable W / O emulsion.
[0036] 1.3 Put the organic solvent on a rotary evaporator (N-1001...
Embodiment 2
[0046] 1.5 Homogenize the prepared liposome suspension through a high-pressure homogenizer (Em-C3, Avestin, Canada) (15000psi, 3 cycles), and the obtained positive liposome particle size distribution of the encapsulating contrast agent 100-200nm.
[0047] All the other steps are the same as in Example 1.
Embodiment 3
[0049] 1.1 Accurately weigh 20 mg of dipalmitoylphosphatidylethanolamine (DOPC, NOF Corporation), 30 mg of 3β-[N-(N', N'-dimethylaminoethyl)] carbamoyl-cholesterol (DC-Chol) Add 8ml of chloroform / diethyl ether (4, V / V) to a 250ml round bottom flask to dissolve it completely.
[0050] 1.2 Dilute ferric oxide tartaric acid solution with water for injection to 3 mg / ml in a cell pulverizer and sonicate (400w, 5s each time, 40 times) to make it evenly dispersed, take 1.5ml into the above round bottom flask, and ultrasonicate in a water bath 30min to form a stable W / O emulsion.
[0051] 1.4 Add 10ml of phosphate buffer (pH 7.4) and vortex to make the gel fall off. Put it on the rotary evaporator and continue rotary evaporation for 2 hours until the liposome suspension is formed.
[0052] All the other steps are the same as in Example 1.
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Abstract
The present invention relates to the technical field of the biological medicine. The present invention particularly relates to a targeted preparation comprising a liposome with the encapsulated contrast agent and the nucleic acid, and the present invention also comprises the preparation method, which is applied in diagnosis and therapy. The targeted preparation of the present invention can be applied in treatment and diagnosis for the human and animal diseases. After preparation of the liposome with the encapsulated contrast agent, the liposome suspension and the nucleic acid are cultivated at room temperature for 15 minutes to 24 hours. After sterilization, the targeted preparation is obtained. Measurement results comprising particle diameter distribution, electric microscope observationand zeta potential all demonstrate that the targeted preparation of the present invention possesses stable structure, no toxicity, safe degradation, which completely accords with requirement for the targeted vector. In the present invention the targeted preparation carrying the Apo-A-I gene or the SR-BI gene are obtained through preparation. Genetic therapy and the treatment effect detection are conducted on rats by using the preparation. The result displays that the targeted preparation has good genetic therapy effect and high safety property.
Description
technical field [0001] The invention relates to the technical field of biomedicine, in particular to a targeted preparation consisting of liposomes encapsulating contrast agents and nucleic acids, which can be used for diagnosis or treatment, and a preparation method thereof. Background technique [0002] With the development of modern molecular biology, human beings have obtained an unprecedented means of treating diseases, that is, gene therapy. The basic idea is to introduce normal genetic material into the patient's somatic cells and express it to produce therapeutic proteins, so as to achieve purpose of treating disease. A key step in gene therapy is the establishment of a gene delivery system. The target gene needs to be transfected into target cells with the help of a carrier—the gene delivery system. At present, liposome-mediated gene transfer is widely used in gene therapy, in which positive liposomes (also known as cationic liposomes) have low toxicity, low immuno...
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Patent Type & Authority Patents(China)
IPC IPC(8): A61K9/127A61K48/00A61K49/06A61P43/00
Inventor 陈建明陆建平郑肖利吴轶娜李莹何新红郭丹沈碧霞张悦马小龙张扬
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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