27 results about "Mrna transfection" patented technology

The invention relates to a preparation method and application of a metal ion type lipid nanoparticle delivery system for regulating mRNA transfection. The method comprises the following steps: (1) mixing mRNA and metal ions in a solvent for 5-15 minutes to obtain a mixture solution (water phase); and (2) blowing, beating and mixing the nanoparticle-containing solvent obtained in the step (1) and a lipid solvent, and wrapping the mixture in the water phase with lipid nanoparticles to obtain the metal ion type lipid nanoparticle delivery system for regulating mRNA transfection. The preparation method has the characteristics of mild preparation conditions and simple preparation steps. According to the application, the organ specific transfection capacity of a lipid nanoparticle delivery system containing different metal ions is verified through mouse experiments. An in-vitro imaging result shows that the system has an enhancement or inhibition effect on mRNA transfection in different organs. The discovery provides a reference basis for metal ion selection for clinically and accurately regulating mRNA in-vivo transfection, and has an important guiding value.

Provided are a new cationic lipid compound, and a preparation method therefor, a composition thereof and the use thereof. The cationic lipid has a structure as represented by formula I. Lipid nanoparticles prepared from the cationic lipid have a stable nanostructure, have a relatively narrow size distribution, and can be stored for a relatively long time at a low temperature. Moreover, the lipid nanoparticles have relatively good biocompatibility and a relatively high in-vivo mRNA transfection efficiency. The core structures of the cationic lipids can all be constructed by means of a Ugi reaction. The reaction involves simple operation and mild reaction conditions, does not require additional catalysts, has high atomic economy, is simple and easy to carry out, and incorporates inexpensive and readily available raw materials, such that not only is higher safety provided, but the industrial production and quality control of the cationic lipids are also facilitated. The new cationic lipid has good application prospects.

The invention belongs to the technical field of biological medicine, and particularly relates to ionizable lipid and a preparation method and application thereof, a drug carrier and a drug composition. According to the ionizable lipid provided by the invention, hydroxyl is introduced to the head, meanwhile, the overall structure is similar to a cone, the head is small, the tail is large, and through structural improvement cooperation, lipid nanoparticles prepared from the ionizable lipid have higher in-vivo mRNA transfection efficiency, and an unexpected technical effect is achieved. The ionizable lipid provided by the invention has a nitrogen branching structure, and the amino group at the branching center is easy to protonate under the physiological pH condition to form part of positive charges, so that the ionizable lipid can be combined with nucleic acid with negative charges, the loading rate of nucleic acid drugs is improved, and the ionizable lipid is suitable for preparing drug carriers or pharmaceutical compositions.

The invention relates to the technical field of drug delivery, in particular to an ionizable lipid compound and application thereof. The invention provides an ionizable lipid compound simultaneously containing # imgabs0 # fragments. The chemical structure of the ionizable lipid compound is shown as a formula (I). The invention also provides an application of the ionizable lipid compound. The mRNA-LNP containing the ionizable lipid compound provided by the invention can integrally improve the biocompatibility of the LNP and the mRNA transfection efficiency, can efficiently deliver a chimeric antigen receptor (CAR) to cells which are extremely difficult to transfect, such as T cells, NK cells, macrophages and the like, especially when being applied to the field of cell therapy, and has an unexpected technical effect.

The invention provides a targeted mRNA delivery system of muscle tissues and / or muscle cells based on aptamer functionalized LNP (Apt-LNP), the delivery system can significantly improve mRNA transfection of the muscle tissues, and meanwhile, mRNA expression in main organs such as liver and spleen is greatly reduced. The expression intensity of muscle tissue in-situ mRNA is improved through modification of the aptamer, meanwhile, mRNA expression in main organs such as the liver and the spleen is greatly reduced, and side effects caused by off-target of mRNA infectious disease vaccines are expected to be avoided. The method is expected to be applied to the field of infectious disease mRNA vaccines The invention also provides a targeting mRNA delivery system of lymph node tissues and / or immune cells based on the aptamer functionalized LNP (Apt-LNP). Compared with an existing lymph node targeting system, the Apt-LNP modified by the aptamer not only can realize lymph node mRNA targeting, but also can further target immune cells, and is expected to be applied to the field of tumor mRNA vaccines to enhance the curative effect of tumor vaccines.

A cationic lipid compound, and a preparation method therefor and use thereof are provided. The cationic lipid compound features a hydroxyl group at the head part, and its overall structure resembles a cone with a small head and a large tail. The LNPs prepared using the cationic lipid compounds with the aforementioned optimal structure usually exhibit enhanced biocompatibility and higher in vivo mRNA transfection efficiency, achieving unexpected technical effects. The synthesis route of the cationic lipid compounds is straightforward and practicable, with inexpensive and readily available raw materials, facilitating industrial production. Furthermore, the LNPs produced from the cationic lipid compounds possess a stable nanostructure that can be stored at low temperatures for a long time, thereby prolonging the shelf life of the pharmaceutical products while reducing the transportation requirements.

This invention belongs to the field of biomedical technology, specifically relating to a delivery system for type III collagen mRNA and its applications. The nucleic acid delivery system of this invention includes a delivery carrier and a delivered nucleic acid, wherein the nucleic acid is collagen mRNA, and the delivery carrier includes one or more of modified amino acids, growth factors, and polymeric materials. Through intradermal injection, the delivery system of this invention can be administered to the dermal layer of the skin and enter relevant cells, utilizing endogenous cellular proteins to achieve mRNA transfection, forming natural human type III collagen. This invention avoids the immunogenicity risks and structural inactivation problems associated with exogenous collagen injections, and can efficiently and safely replenish non-regenerative type III collagen in tissues. It can be used for various indications such as treating skin aging, promoting wound healing, reducing scar formation, and improving vascular elasticity.

The invention discloses a linear poly (beta-amino ester) compound loaded with hepatocyte growth factors as well as a preparation method and application of the linear poly (beta-amino ester) compound, and belongs to the technical field of biological medicines. The compound is obtained by self-assembly of linear poly beta-amino ester DD90-C12-DMEN and HGF mRNA in a buffer solution, the compound is suitable for aerosol inhalation delivery, higher mRNA transfection efficiency is obtained in lung-related epithelial / stromal cells, macrophages and other cells difficult to transfect, and particularly, the compound shows remarkable transfection advantages on the macrophages.

Leveraging the extensive surface area of the lungs for gene therapy, inhalation route offers distinct advantages for delivery. Clinical nebulizers that employ vibrating mesh technology are the standard choice for converting liquid medicines into aerosols. However, they have limitations when it comes to delivering mRNA through inhalation, including severe damage to nanoparticles due to shearing forces. A microfluidic aerosolization platform (MAP) can preserves the structural and physicochemical integrity of lipid nanoparticles, enabling safe and efficient mRNA delivery to the respiratory system. Results demonstrated the superiority of the novel microfluidic aerosolizer over the conventional vibrating mesh nebulizer, as it avoided problems such as particle aggregation, loss of mRNA encapsulation, and deformation of nanoparticle morphology. Notably, aerosolized nanoparticles generated by the microfluidic aerosolization platform led to enhanced transfection efficiency across various cell lines. In vivo experiments with mice that inhaled these aerosolized nanoparticles revealed successful, lung-specific mRNA transfection without observable signs of toxicity. This pioneering MAP represents a significant advancement for the pulmonary gene therapy, enabling precise and effective delivery of aerosolized nanoparticles.

The present invention relates to the technical field of biochemistry, molecular biology and cell biology, and provides an amyloid fusion protein and a preparation method and application thereof. The amyloid fusion protein has a connected amyloid accumulation core fragment and a target protein specific binding fragment, and can specifically recruit the target protein in the cell through the inclusion body formed by the accumulation of the fusion protein, and specifically reduce the availability of the target protein in the cell to interfere with the normal biological function thereof. The design concept of the amyloid fusion protein of the present invention can be applied to the research and development of targeted transcription factor drugs and the design of amyloid fusion protein-related drugs using mRNA transfection or protein delivery methods.

The invention discloses a guanidyl lipid compound and application thereof, and belongs to the technical field of medicines. The structural general formula of the compound is as shown in any one of formula (I) to formula (IV), and guanidyl of the molecule is helpful to be combined with mRNA and other nucleic acid drugs and assists nucleic acid molecules in entering cells to play a role. The invention discloses various applications of the guanidyl lipid molecule. On one hand, the molecules can be used as a transfection enhancer to be added into four-component lipid nanoparticles in a small amount, so that the in-vivo and in-vitro mRNA transfection effects of the original lipid nanoparticles are enhanced; on the other hand, the reagent can be used as a single-component transfection reagent to directly deliver mRNA into cells to realize mRNA transfection. Under the applications, the guanidyl lipid molecules show relatively good nucleic acid delivery and transfection capacities, and have important significance in promoting the development of nucleic acid delivery.

According to the preparation method of the controllable gradient scaffold and the controllable gradient scaffold, the cells are modified through mRNA delivery, the modified cells are loaded to the specific position to construct the gradient scaffold, the seed cells are functionalized through mRNA transfection, and therefore the seed cells are functionalized, and the preparation method of the controllable gradient scaffold has the advantages that the preparation method is simple and convenient, and the preparation cost is low. Compared with the traditional recombinant protein, plasmid, metal ion and other modes, the method has better effectiveness, safety and clinical transformation potential; meanwhile, the used mRNA has extremely high designability, and functional factors required by coding can be designed according to regeneration requirements of different tissues, so that a specific shape and an internal structure are designed according to clinical requirements, personalized customization of the structure is realized, and the requirements of accurately inducing BMSCs to differentiate into cartilage, hypertrophic cartilage and osteogenesis in a three-dimensional space of the stent are better met.

To provide mRNA transfection of immune cells.SOLUTION: The present disclosure relates to methods of modifying immune cells by delivering a modified messenger RNA (mRNA) encoding a chimeric antigen receptor (CAR) and modified immune cells comprising the CAR. The present disclosure encompasses, inter alia, methods, systems, and compositions for modifying immune cells, including monocytes, macrophages, and / or dendritic cells. In some embodiments, the provided methods, systems, and / or compositions provide for enhanced production and / or enhanced properties of modified immune cells.SELECTED DRAWING: Figure 1A

The present invention relates to the technical field of biopharmaceutics and vaccines, and in particular, to an aluminum-based self-assembled delivery system for mRNA, a method for preparing same, and use thereof. The aluminum-based self-assembled delivery system for messenger RNA is a nanocrystallite formed by in-situ self-assembly of an aluminum ion, an mRNA, and a co-assembled molecule driven by coordination. The delivery system serves as a vector-free self-delivery system serving for interleukin 21 mRNA, and can achieve efficient delivery of interleukin 21 mRNA and improved mRNA stability and targeted mRNA delivery efficiency. The nanoparticle of the present invention features good stability in serum and low toxicity, and can achieve efficient mRNA transfection in melanoma cells and significant inhibition of tumor growth in a melanoma mouse model. Therefore, the present invention provides an efficient and convenient preparation method for nucleic acid delivery systems, and has extremely high practical value.

The present disclosure provides compounds of the formulae herein (e.g., Formula (I), Formula (I')), and pharmaceutically acceptable salts thereof, and compositions and kits comprising the compounds, or pharmaceutically acceptable salts thereof, which are useful for mRNA delivery to immune cells (e.g., tumor-infiltrating monocytes (TIMs), myeloid-derived suppressor cells (MDSCs)). The present disclosure also provides methods of treating or preventing diseases or disorders (e.g., diseases or disorders associated with TIMs, MDSCs) by administering the compounds, or pharmaceutically acceptable salts thereof, or compositions thereof, to a subject in need thereof, and methods of preparing the compounds, or pharmaceutically acceptable salts thereof.

The invention provides a preparation method and application of lung target organ to branched poly (beta-amino ester) group mRNA (messenger Ribonucleic Acid) compound nanoparticles. Linear low-molecular-weight poly (beta-amino ester) is used as an arm, a diamine monomer is used as a core molecule, and a brand-new-structure lung target organ-branched poly (beta-amino ester) is obtained. The compound nanoparticles formed by the lung organ targeted branched poly (beta-amino ester) and mRNA (messenger ribonucleic acid) are relatively uniform in distribution, good in stability, biodegradable and good in biocompatibility. An in-vitro transfection result shows that the compound nanoparticles formed by the lung organ targeted branched poly (beta-amino ester) and the mRNA can efficiently deliver the mRNA in various tissue cell lines. An in-vivo transfection result shows that the functional branched poly (beta-amino ester) prepared by the invention can safely and efficiently mediate mRNA specific targeting lung tissues, and shows extremely high mRNA transfection efficiency, so that an application basis is provided for lung cancer gene therapy and vaccine development.

The present invention relates to the technical fields of biopharmaceuticals and vaccines, in particular to an aluminum-based self-assembly delivery system for mRNA, a method for preparing it, and its use. The aluminum-based self-assembly delivery system for messenger RNA is a nanocrystal formed by in situ self-assembly of aluminum ions, mRNA, and co-assembly molecules driven by coordination. This delivery system can function as a vector-free self-delivery system for delivering interleukin 21 mRNA, and can achieve efficient delivery of interleukin 21 mRNA, improved stability of mRNA, and improved delivery efficiency of target mRNA. The nanoparticles of the present invention are characterized by good stability in serum and low toxicity, and can achieve efficient mRNA transfection into melanoma cells and significant inhibition of tumor growth in melanoma mouse models. Therefore, the present invention provides an efficient and convenient preparation method for a nucleic acid delivery system, which has extremely high practical value.

The invention provides a novel cationic lipid compound as well as a preparation method, a composition and application thereof. The cationic lipid provided by the invention has a structure as shown in a formula I. The prepared lipid nanoparticles have stable nanostructures and narrow size distribution, and can be stored at low temperature for a long time; in addition, good biocompatibility and high in-vivo mRNA (messenger ribonucleic acid) transfection efficiency are realized. The core structure of the cationic lipid can be constructed through Ugi reaction, the reaction operation is simple, the reaction condition is mild, no additional catalyst is needed, the atom economy is high, the method is simple and feasible, the raw materials are cheap and easy to obtain, the safety is higher, the industrial production and quality control are facilitated, and the method is suitable for industrial production. The novel cationic lipid has a good application prospect. # imgabs0 #

Lipid nanoparticles (LNPs) are widely used for RNA delivery but are limited by inefficient RNA release following endosomal escape. Disclosed herein are hybrid polymer-lipid nanoparticles (PLNPs) incorporating acid-responsive poly(lactic acid)-block-poly(carboxybetaine) zwitterionic polymers to enhance RNA delivery efficiency. The polymers are cationic at physiological pH to enable RNA complexation but become neutral at endosomal pH, reducing RNA binding affinity and promoting release. These polymers were integrated into clinically approved LNP formulations to form PLNPs. The resulting PLNPs showed up to a 5.4-fold decrease in siRNA IC50 values and a 4-fold increase in mRNA transfection across multiple cell lines. Enhanced cytosolic RNA levels were confirmed via confocal microscopy, with uptake and endosomal escape comparable to standard LNPs. The improvement in transfection efficiency was lost when acid-inert polymers were used, confirming the role of the acid-responsive polymers. This approach provides a versatile platform to improve RNA delivery from existing LNP systems.

The invention discloses a glycoside sterol component lipid nanoparticle for regulating and controlling mRNA transfection as well as a preparation method and application of the glycoside sterol component lipid nanoparticle, and belongs to the technical field of pharmaceutical preparation preparation. The nanoparticle is prepared from DLin-MC3-DMA, DOPE, different glycosides or sterols and DSPE-PEG-GSH. The different glycosides or sterols comprise amygdalin Amy, beta-sitosterol, stigmasterol Sti, ginsenoside Rg1 and salidroside Sal. Stigmasterol can improve enrichment of LNP in the spleen, ginsenoside can adjust immunity and enhance the anti-tumor immune effect, the preparation method is simple, needed equipment is conventional equipment, large-scale production can be achieved, and therefore the stigmasterol and ginsenoside compound has good industrial application value and market prospects.

The invention discloses an electrotransfection buffer solution suitable for long-fragment mRNA transfection cells as well as a preparation method and application of the electrotransfection buffer solution. The electrotransfection buffer solution is prepared from the following components with final concentration: 100 to 150mmol / L of sodium chloride, 1 to 5mmol / L of potassium chloride, 5 to 15mmol / L of disodium hydrogen phosphate, 1 to 3mmol / L of monopotassium phosphate, 0.001 to 0.2 mmol / L of glucose, 1 to 15mmol / L of magnesium chloride, 0.1 to 1.0 mmol / L of calcium chloride and the balance of water. The electroporation buffer solution is low in permeability and conductivity, and high electroporation efficiency can be achieved; through comparison, the transfection efficiency of the electrotransfection buffer solution is obviously higher than that of a commercial electrotransfection buffer solution and a complete culture solution under the condition of the same dosage, and the electrotransfection buffer solution has practical application value.

The invention discloses an imidazole ionizable lipid and a preparation method and application of metal coordination lipid nanoparticles of the imidazole ionizable lipid. Specifically, 32 kinds of imidazole ionizable lipids are designed and synthesized, and BR5-2 LNPs with the optimal mRNA delivery performance is obtained through screening. On the basis of the coordination effect of imidazole and Zn, the metal coordination lipid nanoparticles Zn-BR5-2 NAs are constructed. An XPS result proves that Zn and imidazole nitrogen are coordinated. The nanoparticles have excellent performance in the aspects of mRNA transfection efficiency, cellular uptake and lysosome escape, and can significantly promote the dendritic cell maturation and antigen presentation capability. In-vivo experiments show that the Zn-BR5-2 NAs has a good spleen targeting property. In a B16F10-OVA melanoma model, the Zn-BR5-2 NAs loaded with OVA mRNA (messenger Ribonucleic Acid) realizes a remarkable tumor prevention and treatment effect. According to the mRNA vaccine based on Zn-BR5-2 NAs, the anti-tumor effect is remarkably improved by enhancing immune response, an effective strategy is provided for tumor immunotherapy, and the mRNA vaccine based on Zn-BR5-2 NAs has wide application prospects.

The invention relates to the technical field of gene engineering, in particular to coding nucleic acid of Cre recombinase and application of the coding nucleic acid. The Cre recombinase is subjected to codon optimization of four versions, the protein expression quantity of the obtained Cre recombinase is remarkably improved compared with that of a wild type and other contrasts, the mRNA transfection reagent prepared from the nucleic acids can have good transfection efficiency, and the effect of a Cre recombinase system can be more effectively played when the mRNA transfection reagent is transferred into cells, particularly 293T cells.

The invention discloses a rapid preparation method and application of chimeric antigen receptor T cells, and belongs to the technical field of biological medicine. According to the method, the traditional T cell activation stage is completely avoided, and the CAR-T cell with high expression of CAR similar to the conventional T cell activation is rapidly obtained in a manner of directly co-culturing the separated T cell and nicotinamide adenine dinucleotide in vitro for a short time and then transfecting mRNA (messenger Ribonucleic Acid). The traditional T cell activation stage is skipped, the preparation time of the CAR-T cells is greatly shortened, and the cost is reduced; in-vitro co-culture with NAD < + > is utilized, so that the T cell activity and the mRNA transfection efficiency are improved; by adopting lipid nanoparticles or electrotransfection of mRNA, the potential risk of a viral vector is avoided, and the safety of the preparation process is improved; the prepared CAR-T cell has the characteristic of high expression of CAR and relatively strong in-vitro tumor cell killing activity, and is expected to improve the clinical treatment effect.

The invention provides a preparation method and application of an in-vivo CAR-T cell for treating interstitial lung disease, and provides a T cell of a chimeric antigen receptor which is modified by genetic engineering and is used for expressing targeted fibroblast activating protein. A targeted fibroblast activation protein (FAP) CAR-T cell therapy model is constructed by targeting CD5 entrapped mRNA nano-liposome (LNP) transfection and lentiviral vector, and the method comprises the following steps: firstly, verifying the difference of the killing ability of FAP CAR-T constructed by CD5 LNP-mRNA and lentivirus in a 293T cell which stably co-expresses FAP, Luciferase and mCherry, and then verifying the difference of the killing ability of FAP CAR-T constructed by LNP-mRNA and lentivirus in the 293T cell which stably co-expresses FAP, Luciferase and mCherry; further verifying the effectiveness of the CAR-T cells constructed by transfecting the CD5 LNP-mRNA on cell lines of human fibroblasts (CDD19Lu, LL29 and LL97A) and primary fibroblasts of human and mice in vitro, and further verifying the effectiveness and safety of the CD5 LNP-FAP CAR-T in treatment of pulmonary fibrosis through in-vivo experiments in animals. A novel and effective anti-fibrosis treatment thought is provided for patients with fibrosis interstitial lung diseases.

The invention belongs to the technical field of biological medicines, and discloses a tertiary amine oxide derived zwitterionic lipid and application thereof. According to the preparation method, four components Ugi of aldehyde, isonitrile, amine and carboxylic acid are subjected to a reaction to synthesize ionizable cationic lipid, and then a tertiary amine head group of the ionizable cationic lipid is subjected to an oxidation reaction to obtain the tertiary amine oxide derived zwitterionic lipid. The tertiary amine oxide derived zwitterionic lipid disclosed by the invention can replace auxiliary phospholipids to prepare lipid nanoparticles and is used for nucleic acid drug delivery; the muscle in-situ mRNA transfection efficiency can be improved, the liver off-target protein expression level can be reduced, and the application requirements of nucleic acid drugs, nucleic acid vaccines, gene editing and the like can be met.

Leveraging the extensive surface area of the lungs for gene therapy, inhalation route offers distinct advantages for delivery. Clinical nebulizers that employ vibrating mesh technology are the standard choice for converting liquid medicines into aerosols. However, they have limitations when it comes to delivering mRNA through inhalation, including severe damage to nanoparticles due to shearing forces. A microfluidic aerosolization platform (MAP) can preserves the structural and physicochemical integrity of lipid nanoparticles, enabling safe and efficient mRNA delivery to the respiratory system. Results demonstrated the superiority of the novel microfluidic aerosolizer over the conventional vibrating mesh nebulizer, as it avoided problems such as particle aggregation, loss of mRNA encapsulation, and deformation of nanoparticle morphology. Notably, aerosolized nanoparticles generated by the microfluidic aerosolization platform led to enhanced transfection efficiency across various cell lines. In vivo experiments with mice that inhaled these aerosolized nanoparticles revealed successful, lung-specific mRNA transfection without observable signs of toxicity. This pioneering MAP represents a significant advancement for the pulmonary gene therapy, enabling precise and effective delivery of aerosolized nanoparticles.