45results about How to "Improve transfection efficiency" patented technology

The present invention relates to novel compositions and methods for delivering substances to target tissues and cells by contacting the targets with delivery systems associated with membranes (e.g., biocompatible or bioerodable membranes). More particularly, the present invention is directed to dendrimer-based methods and compositions for use in disease therapies, wound healing, and generally, improved gene transfection and compound delivery to target cells and tissues in vitro and in vivo.

The present invention provides a self-assembling peptide system which utilizes a bioactive sequence which enhances transfection efficiency. In particular, the present invention provides compositions and methods for transfecting aggregates of cells at a higher efficiency.

The invention features selected polymers such as poloxamer 188 (Pluronic® F68), poloxamer 237 (Pluronic® F87), poloxamer 401 (Pluronic® L121), poloxamer 338 (Pluronic® F108), Poloxamer 124 (Pluronic® L44), Poloxamer 184 (L-64), and poloxamines (Tetronics® 904, 908, 1107, and 90R4) that enhance expression from nucleic acids when administered into an organism, in particular to muscle, and further provides selected poloxamer formulations for delivery of nucleic acids to the liver.

The invention relates to a hollow mesoporous silicon dioxide nanoparticle and a preparation method thereof. The preparation method comprises the following steps: magnetically stirring and mixing ethanol, deionized water and ammonia water at a certain temperature, adding ethyl orthosilicate for continuous reaction, adding premixed ethyl orthosilicate and trimethoxyoctadecylsilane for continuous reaction, vacuum drying an obtained mixture after being etched by sodium carbonate, calcining at 550 DEG C to obtain the hollow mesoporous silicon dioxide nanoparticle. The nanoparticle is dispersed in ultrapure water, and polymine is added to obtain a hollow mesoporous silicon dioxide nano-carrier after mixing; and the hollow mesoporous silicon dioxide nano-carrier is mixed with a gene to obtain a hollow mesoporous silicon dioxide gene nano-carrier. The hollow mesoporous silicon dioxide nano-carrier prepared by the preparation method disclosed by the invention has the advantages of good dispersity, high gene loading capacity and high transfection efficiency (twice of 25kDa PEI), and has a practical value of clinical application.

The invention discloses a composite siRNA nanocarrier, comprising exosome lipid membrane from autologous tumor cells and siRNA-carrying cationic serum albumin covered in the exosome lipid membrane. The invention also discloses a preparation method and application of the composite siRNA nanocarrier. The composite siRNA nanocarrier has the advantages of high transfection, specific targeting propertyof cancer metastatic tissues, good biocompatibility, good stability, zero immunogenicity and the like.

The invention discloses a ternary complex nanodrug, a preparation method and application thereof in preparation of a light controlled release nano-delivery system. The ternary complex nanodrug comprises a thioketal bond crosslinked low molecular weight PEI-based polymer, a hyaluronic acid polymer attached with a photosensitizer, and a nucleic acid drug. The photosensitizer contained in the complexcan generate reactive oxygen species (ROS) under the control of exogenous red light, realize photochemical internalization (PCI) mediated lysosome escape and photopromoted polymer degradation, thus finally realizing the purposes of reducing the toxic and side effects of materials, improving the transfection efficiency, and enhancing the tumor gene therapeutic effect.

The invention relates to the field of polymer modification, and in particular to a modified polyethyleneimine and application thereof in the preparation of a gene transfection vector reagent. According to the present invention, hydrophobic rosin acid or dehydroabietic acid is grafted onto polyethyleneimine to obtain the modified polyethyleneimine. The hydrophobic group of the modified polyethyleneimine forms a dense hydrophobic core with strong hydrophobicity due to the intermolecular forces in water, hydrophilic polyethyleneimine forms hydrophilic shell, thereby forming a nano micelles with shell-core structure, and nanoparticles with small particle size formed after DNA binding. The composite nanoparticles in smaller size gains tighter structure, so that composite nanoparticles are easier to pass through the cell membrane structure of a lipid bilayer, and the efficiency of transfection is improved. The modified polyethyleneimine can be applied to the preparation of gene transfection vectors and has the characteristics of high biological safety, high transfection efficiency, and low cost.

The present invention provides gold nanoparticle-liposome composite nanoparticles, which are gold nanoparticles encapsulated with a liposome vesicle, wherein the surface of the liposome vesicle is modified with distearoyl phosphatidylethanolamine-polyethylene glycol, the surface of the gold nanoparticles is modified with plasmid and a penetrating peptide, and the liposome vesicle comprises dioleoyl phosphoethanolamine, 2,3-dioleyloxypropyltrimethyl ammonium chloride and cholesterol. The invention further provides a preparation method of the gold nanoparticle-liposome composite nanoparticles ,and a pharmaceutical composition comprising the gold nanoparticle-liposome composite nanoparticles. According to the present invention, the gold nanoparticle-liposome composite nanoparticles as the carrier have the high transfection effect, and the pharmaceutical composition has good treatment effects on various tumors.

The invention discloses a modified polyethyleneimine derivative as well as a synthesis method and application thereof. According to the invention, Triton X-100 is conjugated to polyethyleneimine, suchthat a PEI-Triton-N carrier is obtained. In-vitro cell evaluation shows that the carrier shows good safety and stability; cell transfection effect experiments show that the carrier shows efficient and low-toxicity gene delivery capacity, can significantly improve the gene transfection efficiency of various cells including skin immortalized cells HaCaT difficult to transfect, and is an excellent in-vitro transfection reagent; an in-vivo toxicity experiment shows that the carrier has no obvious systemic toxicity; and a gene delivery experiment result of mouse skin local administration shows that the carrier has good local administration gene delivery capability and is a good local administration gene delivery vector.

The invention provides an alpha helix cationic polypeptide as well as a preparation method and application of the alpha helix cationic polypeptide. The alpha helix cationic polypeptide takes a photosensitizer porphyrin as a core and alpha helix polypeptides as four arms, and a nucleotide drug prepared from the polymer can be applied to a nucleotide drug delivery system. The polymer provided by theinvention has good biocompatibility, low cytotoxicity and light activation ability. The polymer provided by the invention is self-assembled in a water solution to form nanoparticles which has good stability, biocompatibility and low cytotoxicity; the preparation method is simple and high in repeatability, and the polymer serving as a carrier is not only capable of protecting nucleic acid such asDNA from being degraded, but also capable of being combined with the scale effect of the nanoparticles so as to be used for treating diseases; and in addition, the transfection promoting effect for different cancer cells can be achieved by virtue of the light activation ability.

The invention discloses a magnetic nanometer gene vector system as well as a preparation method and an application thereof. The gene vector system comprises electronegative magnetic nanoparticles, positive-ion vectors and genes, wherein the three components are prepared into the magnetic nanometer gene vector system according to certain proportions and composite sequences through electrostatic interactions. The magnetic nanometer gene vector system disclosed by the invention is simple in preparation method and has physicochemical properties of small grain diameter and proper surface charge. Compared with non-magnetic nanometer composites, the magnetic nanometer gene vector system has the advantages that the transfection efficiency to cells of the system is remarkably improved under the action of an additional magnetic field; and meanwhile, the cytotoxicity of the system is further reduced.

The invention discloses an antiviral small nucleic acid and the application of the small nucleic acid in preparing a medicine for suppressing herpes simplex virus (HSV) and human papilloma virus (HPV). The small nucleic acid is a double strand nucleic acid, the positive-sense strand sequence is 5 in a sequence table, and the antisense strand sequence is 6 in the sequence table; the small nucleic acid can be prepared into a temperature-sensitive type gel preparation; the temperature-sensitive type gel preparation is injected (or sprayed) into irregular cavities such as vagina or mouth, the expression of host B2M genes in epithelial cells at the cavities and the cervix uteri and the expression of externalHPV E7 are suppressed, so that the antiviral small nucleic acid can be used for suppressing the infection of the HPV of the HSV-1 virus and/or for treating the precancerous lesions of uterine cervix caused by the HVP.

The invention relates to the technical field of biological medicine, and particularly discloses a lipid compound, a composition containing the same and application. The lipid compound has a structure as shown in a formula (I) or a formula (II). The lipid compound can be ionized into a cationic compound under an acidic condition, and the cationic compound is combined with negatively-charged active pharmaceutical ingredients through electrostatic interaction, so that drug-loaded lipid nanoparticles are assembled, and the active pharmaceutical ingredients are delivered. The lipid compound provided by the invention is simple in structure, simple in reaction path and high in yield, and the constructed drug-loaded lipid nanoparticles can be used for preparing nucleic acid drugs, gene vaccines, polypeptide or protein drugs and small molecule drugs, and have wide application prospects.

The invention discloses a method for transfecting an exogenous nucleic acid by use of atmospheric pressure cold plasma jet and belongs to the technical field of biology. The method comprises the following steps: 1) counting to-be-transfected cells, planting the cells in a porous cell culture plate, and adding a culture solution to obtain a to-be-transfected cell solution; 2) adjusting a plasma jet source device, introducing a basic gas, applying sinusoidal high voltage after the gas flow is stable, and keeping the voltage constant until bluish violet plasma jet is blown out from the bottom of a quartz tube; 3) treating the to-be-transfected cells for 5-60 seconds directly under the plasma jet source, moving away the cells and adding the to-be-transfected exogenous nucleic acid, mixing evenly and then putting the mixed solution back into an incubator for continuous culture. The method for transfecting the exogenous nucleic acid by use of the atmospheric pressure cold plasma jet is capable of conveniently transfecting the exogenous nucleic acid; after transfection, the cell mortality rate is relatively lower than those of methods by use of lipidosome, electrotransfection and the like. Besides, the transfection time is very short and the transfection can be completed within dozens of seconds.

The invention provides PLNPs. The PLNPs are nanoparticles wrapped by liposome vesicles, and the surfaces of the liposome vesicles are modified with distearoyl phosphatidylethanolamine-PEG; the nanoparticles are prepared from lipid particles, chondroitin sulfate and protamine; the liposome vesicles are prepared from dioleoyl phosphatidylethanolamine, N-[1-(2,3-dioleyloxy) propyl]-N,N,N-trimethylammonium chloride and cholesterol. The invention further provides a preparation method of the PLNPs and pharmaceutical composition containing the PLNPs. The PLNPs as a carrier have higher transfection effect. The pharmaceutical composition has better treatment effect on various tumors.

The invention discloses a device for ultrasound microbubble-mediated transfection. The device comprises a liquid storage container, a floating board, an ultrasonic transducer and a culture dish, wherein the liquid storage container is used for containing an ultrasound contrast agent, the floating board is in the liquid storage container and floats in the ultrasound contrast agent, the ultrasonic transducer is installed on the floating board and suspends in the ultrasound contrast agent under the floating action of the floating board, the culture dish is suspended in the liquid storage container and is above the ultrasonic transducer. According to the device for ultrasound microbubble-mediated transfection, the ultrasonic transducer suspends in the ultrasound contrast agent through the floating board, so that the ultrasonic transducer does not contact or fix with other fixed piece; when ultrasound acts on the ultrasound contrast agent in the liquid storage container, the energy of the ultrasound is directly transmitted to the ultrasound contrast agent and hardly losses, so that the ultrasound contrast agent generates more microbubbles, so as to improve the transfection effect.

The invention relates to a slow-virus vector system and a preparation method thereof, belonging to the field of biotechnology. The slow-virus vector system comprises a shuttle plasmid CSII-EF-MCS-IRES2-Venus, a packaging plasmid pCAG-HIVgp and an envelop protein plasmid pVSV-G-RSV-Rev; a host cell is transfected by the shuttle plasmid CSII-EF-MCS-IRES2-Venus, the packaging plasmid pCAG-HIVgp and the envelop protein plasmid pVSV-G-RSV-Rev in the vector system, namely the plasmids are packaged in the host cell to obtain the slow-virus vector system. Compared with the prior art, the slow-virus vector system and the preparation method thereof provided by the invention have the advantages as follows: (1) the cost is low: by taking the transfection of cells in per 10 centimeters of 10 dishes asexample, the price of the recommended liposome 2000 with the volume of 600 microliters (60 microliters*10) is about 1500 yuan; however, the cost consumed by utilizing the method can be reduced to 0.62 yuan so that the cost can be greatly reduced; (2) the transfecting efficiency is high and the transfecting rate can be more than 80%; and (3) and the valence of the virus is high and can be 5.79*10<9>.

Embodiments of the invention relate to a nano-composite vector for transfection, a nucleic acid nano-drug as well as a preparation method and application of the nano-composite vector and the nucleic acid nano-drug, belonging to the field of biomedicines. The nano-composite vector for transfection comprises a fullerene derivative and a high-molecular polymer with negative charges, wherein the fullerene derivative and the negatively-charged high-molecular polymer are combined through electrostatic interaction and pi-pi interaction, the zeta potential of the nano-composite vector formed by combination is positive; and the nano-composite vector and siRNA are combined through electrostatic interaction and pi-pi interaction to obtain the nucleic acid nano-drug. The nano-composite vector can load siRNA drugs, can assist siRNA in penetrating through cell barriers and improves the transfection performance of siRNA; and the formed nucleic acid nano-drug is stable in structure and good in biocompatibility, can effectively protect siRNA from being degraded by RNA enzyme, and achieves lung transfection of siRNA through respiratory tract inhalation drug delivery, and therefore, the expression of a target gene in a lung tissue can be effectively inhibited.

The invention relates to a magnetic transfection reagent based on supramolecule self-assembly, wherein magnetic spheres prepared through a high temperature decomposition method and six-site fully-substituted alpha-cyclodextrin-PEI600 (CD-PEI) are assembled through self-assembly to form the magnetic transfection reagent. According to the present invention, the preparation method of the magnetic transfection reagent is provided, the structure of the magnetic transfection reagent is characterized, and the biological toxicity and the transfection performance of the magnetic transfection reagent at the cell level are characterized; the preparation method is mild, simple and green environmental protection; and the synthesized magnetic transfection reagent has characteristics of smaller particle size, surface having a lot of positive charges, good siRNA combination ability, low biological toxicity, high transfection efficiency, and good application prospects, and the performance of the magnetic transfection reagent as the siRNA vector is excellent.

The invention relates to a high-molecular polymer for treating tumors and a preparation method and application thereof, and belongs to the technical field of medicines. The preparation method comprises the following steps: synthesizing PEGMA-(RAFT); synthesizing PEGMA-b-PDPA-(RAFT); synthesizing PEGMA-b-PDPA-b-PDMAEMA-(RAFT); desulfurization reaction; and synthesizing the high-molecular polymer ma< 1 >-PMDM. The invention also discloses the high-molecular polymer prepared by the preparation method and used for treating tumors and application of the high-molecular polymer. According to the preparation method, the high-molecular polymer for treating tumors can be prepared very efficiently; the high-molecular polymer can improve the stability of the carrier, improve the cellular uptake capacity and improve the transfection capacity, the composition and the structure of the carrier have the capacity of overcoming the lysosome biological barrier, and the high-molecular polymer can be applied to a targeting drug carrier for treating tumors.