746 results about "Lysosome" patented technology

The invention provides a membrane protein targeted degradation agent based on cell autophagy, the degradation agent has a first structure combined with membrane protein and also has a second structure capable of inducing cell autophagy, and the first structure is connected with the second structure. According to the membrane protein targeted degradation agent based on cell autophagy, the cell autophagy mechanism is utilized for membrane protein degradation for the first time, due to the fact that autophagy is an internal mechanism of various cells, universal induction can be achieved in various cells, and the autophagy does not depend on factors such as lysosome transporter protein and E3 ligase on cell membranes; the method has the characteristics of broad spectrum and higher universality; the design route of direct connection of the first structure and the second structure is simple and easy to realize.

The invention discloses a PEI and targeting peptide modified magnetic nano-carrier and a preparation method and application thereof. The nano-carrier is regular in morphology and uniform in particle size and has good crystallinity and magnetic performance. E5 polypeptide and RGD cyclic peptide on the surface of the nano-carrier are respectively combined with CXCR4 protein and integrin on the surface of a tumor cell, and the nano-carrier and CXCR4 are internalized into lysosome through an integrin-mediated way, so that the target protein CXCR4 is degraded. The PEI in the nano-carrier can load and deliver siRNA into tumor cells to silence the expression of CXCR4 mRNA (messenger Ribonucleic Acid). Through nucleic acid delivery and a protein targeted degradation strategy, the CXCR4 level of tumor cells is jointly reduced, the migration ability of the tumor cells is inhibited, and meanwhile, the anti-tumor effect of M1 type polarization of macrophages is activated.

The invention belongs to the field of biological medicines, and discloses an application of a stem cell extract in lysosome and mitochondrial related diseases, a preparation method of the stem cell extract comprises the following steps: S1) culturing mesenchymal stem cells and creating stress conditions to stimulate the mesenchymal stem cells; and S2) separating and purifying the culture supernatant of the mesenchymal stem cells cultured in the step S1) to obtain the stem cell extract, wherein S21) the culture supernatant is filtered to obtain a filtrate; s22) performing 3KD ultrafiltration concentration on the filtrate to obtain a crude sample; s23) purifying the crude sample by using molecular sieve exclusion chromatography or reversed-phase chromatography to prepare a purified sample; in molecular sieve exclusion chromatography, by taking the volume of the eluent as a reference, collecting the 10th to 40th mL of eluent, namely a purified sample; and S24) taking the purified sample to form a preparation. According to the application, the culture supernatant containing stress stem cell secretions is taken through stress culture of stem cells, and a purification method is combined, so that the prepared preparation can be used for treating lysosome and mitochondrial related diseases.

The invention relates to a bifunctional compound and application thereof in targeted degradation of protein, the compound comprises a target protein binding compound and a cell-penetrating peptide which are connected, and the target protein binding compound is a compound capable of binding with cell membrane protein or extracellular protein or a derivative thereof; the target protein binding compound and the cell-penetrating peptide are connected through a linker or directly connected. According to the invention, it is found for the first time that specific extracellular proteins or cell membrane proteins can enter cells in an endocytosis manner and are degraded through lysosomes by using cell-penetrating peptides and small molecule conjugates. The method disclosed by the invention has the characteristic of delivering bioactive macromolecules through endosome wrapping without depending on a single receptor, and the effect of degrading the target protein by using the cells can be realized as long as specific binding micromolecules of the protein to be degraded can be found and coupled with the cell-penetrating peptide; a widely applicable strategy is provided for degradation of extracellular proteins and cell membrane proteins.

The invention discloses novel application of notoginsenoside Rg1, namely application of the notoginsenoside Rg1 in preparation of a cathepsin D (CTSD) inhibitor, the notoginsenoside Rg1 can inhibit CTSD enzyme activity so as to recover lysosome structure and function, relieve endoplasmic reticulum stress reaction and relieve alpha-syn aggregation and dopaminergic neuron apoptosis, has a remarkable protection effect on a Parkinson's disease model, and can be used for preparing a novel anti-inflammatory drug. The invention provides a new thought and a potential drug basis for targeted intervention of Parkinson's disease. The invention discloses a new mechanism that Rg1 plays a role in neuroprotection through a CTSD-lysosome-endoplasmic reticulum stress pathway.

The invention discloses a preparation method and application of a QS-21-polyamino acid nano vaccine, and belongs to the field of biomedical nano materials. The preparation method comprises the following steps: modifying gamma-PGA with phenylalanine ethyl ester (F) to obtain a polymer PF40 of which the F grafting rate is 40%; aiming at the characteristic that QS-21 is strongly combined with cholesterol, PF40 (PFC) grafted with cholesterol (C) is developed, and PFC wraps OVA and QS-21 through hydrophobic force to construct a nano vaccine OQPFC. The cholesterol on the PFC can reduce the combination of the QS-21 and the cholesterol on a cell membrane so as to reduce the biotoxicity of the QS-21, and the QS-21 loading efficiency of the nano vaccine is improved, so that a better treatment effect is achieved. The OQPFC can escape from lysosome through carboxyl protonation, and a large amount of QS-21 and OVA are released in cytoplasm, so that the synergistic cellular immune activation effect of the carrier adjuvant PFC and the saponin adjuvant QS-21 is realized.

The invention provides a targeting vector and a method for targeting a host cell, the vector comprises a first molecule combined with an endocytosis receptor of a target cell and a second molecule for promoting release of a substance carried by the targeting vector into cytoplasm, when the targeting vector is a virus vector, the first molecule is not a part of a virus envelope protein, and the second molecule is not a part of a virus envelope protein. The second molecules promote endosome escape or lysosome escape of the targeting carrier. According to the present invention, the first molecule is designed according to the endocytosis receptor of the to-be-targeted cell so as to target different types of cells, and after the vector is subjected to reasonable mutation, the infection of the cells not requiring the targeting can be avoided, and the accuracy of the vector can be improved.

Aspects of the disclosure relate to complexes comprising an anti-TfR1 antibody covalently linked (e.g., via a linker such as a peptide linker) to a lysosomal enzyme (e.g., an acid alpha glucosidase enzyme), and methods of making and using the fusion complexes to treat a lysosomal storage disease (e.g., Pompe disease).

The invention discloses application of a lysosome cathepsin L inhibitor in a medicine for treating or relieving allergic airway inflammation, and relates to the field of allergic airway inflammation. It is found through research that inflammatory response of asthma can be relieved by inhibiting expression of lysosome CTSL in macrophages and animal models; the inflammatory response of asthma can be relieved by inhibiting expression of lysosome CTSL in macrophages by using a small-molecule inhibitor and myeloid specific knockout CTSL, so that the lysosome cathepsin L inhibitor can effectively relieve allergic airway inflammation, and a new direction can be provided for research and development of drugs for treating and relieving allergic airway inflammation in the future.

The invention belongs to the field of biological medicines, and relates to a drug-loaded nano-vesicle as well as a preparation method and application thereof. The drug-loaded nano-vesicle comprises a vesicle core and a drug-loaded nano-vesicle, wherein the vesicle core comprises siRNA (small interfering Ribonucleic Acid) capable of specifically targeting and silencing an NR1D1 gene; the vesicle membrane is formed by fusing an erythrocyte membrane, a macrophage membrane, cardiolipin, cholesterol and lecithin. The drug-loaded nano-vesicle can specifically target macrophages in a sepsis immunosuppression stage, has an intracellular response release function, recovers BMAL1 and IGF2BP2-ATP6V1B2 / ATP6V0c axis functions by inhibiting NR1D1 expression, reconstructs a macrophage phagocytosis function and lysosome-dependent bacterium removal capability, and can be used for preparing a drug-loaded nano-vesicle with a specific targeting function. The survival rate of sepsis immunosuppression model animals is obviously improved; and the bacterial load is reduced. Compared with a traditional electroporation method, the preparation method disclosed by the invention has the advantage that the encapsulation efficiency of siRNA is remarkably improved.

The invention discloses a sulfatase-driven polypeptide condensate with a chemotherapy sensitization effect and a preparation method thereof, and belongs to the field of biological medicines. According to the principle that sulfatase can catalyze hydrolysis of sulfate bonds, main polypeptide YSO4F responded by sulfatase enzyme is designed and synthesized; and the enzyme response aggregate m-YSO4F-LSG is blended with a protein G3BP2 ligand FGDF-YSO4F to prepare the enzyme response aggregate m-YSO4F-LSG with a sorafenib chemosensitization effect. Tumor cells can generate stress particles with a chemotherapy drug resistance effect under the stimulation of a chemotherapy drug sorafenib, the treatment effect of the sorafenib is weakened, and the protein G3BP2 is one of core proteins of the stress particles. After the m-YSO4F-LSG enters tumor cells, sulfate groups are hydrolyzed under the catalytic action of sulfatase overexpressed in tumor cell lysosomes, so that phase separation liquid drops d-YF-LSG with high affinity to protein G3BP2 are formed in the cells in situ, the phase separation liquid drops d-YF-LSG act with the protein G3BP2 and are fused with stress particles, chemotherapy drug resistance generated by the stress particles is inhibited, and the chemotherapy effect is improved. The treatment effect is improved.

[Problem] To provide a novel ferroptosis-inducing drug, and in particular, to provide a ferroptosis induction promoter that can effectively promote the induction of ferroptosis even in a cell having ferroptosis resistance even in a ferroptosis-inducing environment (for example, cancer cells and senescent cells). [Solution] Provided is a ferroptosis induction promoter comprising a drug that can reduce lysosomal pH.

The invention discloses a nano degradation agent for targeted degradation of PI3Kalpha protein as well as a preparation method and application of the nano degradation agent. The nano degradation agent comprises a polyamidoamine dendritic polymer G3-NH2, a PI3Kalpha protein targeted receptor and an HSC70 protein targeted receptor, wherein the PI3Kalpha protein targeted receptor and the HSC70 protein targeted receptor are coupled to the surface of the polyamidoamine dendritic polymer G3-NH2. The prepared nano degradation agent enters cells in an endocytosis mode, specifically targets corresponding treatment targets after entering the cells, and then efficiently degrades PI3Kalpha protein through a molecular chaperone-mediated autophagy pathway of lysosome, so that the PI3Kalpha protein can be effectively degraded. The nano degradation agent provided by the invention can be applied to intervention of related diseases caused by PI3Kalpha protein abnormality.

The invention belongs to the technical field of biological medicine, and particularly relates to an LYTAC molecule for targeted degradation of GPC3 and a preparation method and application of the LYTAC molecule. The LYTAC molecule is composed of a target TfR1 nucleic acid aptamer and a target GPC3 nucleic acid aptamer, wherein the nucleotide sequence of the targeted TfR1 nucleic acid aptamer is as shown in SEQ ID NO: 1; the nucleotide sequence of the targeted GPC3 nucleic acid aptamer is as shown in SEQ ID NO: 2. According to the present invention, the research results show that the GPC3 protein content is not affected by the single TfR1 nucleic acid aptamer or the GPC3 nucleic acid aptamer, and the LYTAC molecule can significantly reduce the GPC3 protein content on the surfaces of Hep3B and HepG2 cells. The LYTAC molecule induces GPC3 protein degradation through a lysosome way, then proliferation and migration of liver cancer cells are inhibited, liver cancer treatment is achieved, and the LYTAC molecule has wide application prospects.

The invention discloses a double-selenium lipid nanoparticle as well as a preparation method and application thereof. According to the nanoparticle, a skeleton is composed of cationic lipid, neutral phospholipid, cholesterol and functionalized PEG lipid, meanwhile, mRNA and a TLR agonist for coding therapeutic protein are carried, and the nanoparticle is synthesized through an ethanol injection method; the nanoparticle has both active targeting and strong transfection capabilities, the lipid nanoparticle is actively targeted to a tumor microenvironment and is taken by tumor cells by using biotin modified PEG lipid, and rapid'inclusion-lysosome 'escape is realized in the tumor cells by using trifluorogroup modified PEG lipid. Meanwhile, the nanoparticles are degraded in a glutathione / hydrogen peroxide dual-mode, so that the entrapped medicine is quickly and effectively released; the drug delivery system provided by the invention is simple in synthesis method, high in gene transfection efficiency and good in biological safety, and has potential value in anti-tumor research and clinical application.

The present invention relates to methods and products for assessing autophagic flux, and more specifically provides a biochemical functional measure used to quantify autophagic flux in sub-populations of blood cells. The present invention also provides lysosomal system markers for use in the methods described herein, systems for assessing autophagic flux, methods of identifying markers indicative of autophagic flux, and methods of treating subjects found to be suffering from, or susceptible to, a disease, condition or state associated with autophagic flux dysfunction using the methods described herein.

The invention relates to the cross technical field of biological medicine and image processing, and discloses a mesenchymal stem cell aging detection method based on image processing. The method comprises the following steps: automatically collecting bright field and multi-channel fluorescence images in an integrated cell culture monitoring device; after background correction and illumination homogenization, segmenting the cells by using a U-Net network and extracting single cell contours; calculating characteristics such as cell area, roundness, cytoplasmic ratio, nuclear form irregularity index, lysosome fluorescence intensity mean value and distribution entropy; and inputting a gradient boosting decision tree model to judge the unicellular aging state, and evaluating population aging with a 20% positive rate threshold. According to the invention, label-free, non-invasive and high-flux accurate detection is realized, and the method is superior to manual interpretation.

The present disclosure relates to harnessing endosomal sorting of lipids to enable targeted protein degradation via the lysosomal degradation pathway. Accordingly, provided herein are molecules for degrading target proteins, pharmaceutical compositions of such molecules, and methods of making and using such molecules.

The invention discloses a polypeptide drug chimera and a preparation method thereof, and belongs to the technical field of polypeptide drug carriers. The polypeptide drug chimera is constructed by a recognition segment specifically combined with a target protein and a functional polypeptide carrier mediating the target protein to be transferred to lysosome; an antibody is combined with a lysosome signal amino acid sequence, and pathogenic antigen protein is transferred into a lysosome subcellular organelle for degradation through specific combination of the antibody and different antigens.

The invention provides a carrier material for supporting a protein drug. The carrier material comprises an inner core, a calcium carbonate coating layer coating the inner core and a polyethylene glycol-polyglutamic acid diblock copolymer chain segment anchored on the coating layer. The intracellular protein delivery carrier provided by the invention can effectively support various metal ions and protein drugs with the Fenton / Fenton-like reaction catalysis effect, and after the intracellular protein delivery carrier is endocytosed by cells, the protein drugs, calcium ions and the metal ions with the Fenton / Fenton-like reaction catalysis effect are released; metal ions can catalyze hydrogen peroxide in the endosome / lysosome to generate hydroxyl radicals with high oxidation activity, lipid molecules are oxidized, and a lysosome membrane is destroyed; the released calcium ions can cause mitochondrial calcium overload, improve the level of hydrogen peroxide in cells, promote generation of hydroxyl radicals in endosome / lysosome and damage of membranes of the hydroxyl radicals, induce efficient rupture of the endosome / lysosome and deliver protein drugs into cytoplasm so as to play the functions of the protein drugs.

The invention provides a lysosome extraction method. The lysosome extraction method comprises the following steps: (1) extracting cells; (2) tissue extraction; (3) crude separation of lysosome; and (4) separating the lysosome by magnetic beads. Through EDTA-free digestion optimization, lysis solution and buffer solution formula optimization, a filter membrane interception-magnetic bead purification combined technology, a double-antibody synergistic capture strategy and a competitive mild elution system, the four core problems of low purity, large activity damage, tedious operation and strong equipment dependence in a traditional lysosome separation technology are systematically solved. Experimental data show that the lysosome yield and functional integrity of the method are obviously superior to those of a density gradient centrifugation method, and the method is suitable for trace samples and conventional equipment and has high conversion value. In the future, the technical barrier can be further optimized through automation equipment integration and multi-dimensional function verification, and the application of the method in basic research and clinical diagnosis is promoted.

Aspects of the disclosure relate to bicistronic AAV nucleic acid constructs comprising a transgene encoding hexosaminidase A (HEXA) and hexosaminidase (HEXB) proteins. In some embodiments, the disclosure provides methods for treating or preventing lysosomal storage disorders, such as Tay-Sachs disease and Sandhoff disease, using bicistronic nucleic acid constructs described by the disclosure.

The invention relates to a biological medicine technology, and discloses chloroquine lipid, a preparation method and application thereof, and lipid nanoparticles containing the chloroquine lipid. The chloroquine lipid is synthesized from chloroquine substances and alkylene oxide through a ring-opening reaction, and the chloroquine substances are selected from at least one of chloroquine, hydroxychloroquine, chloroquine derivatives and hydroxychloroquine derivatives. The lipid nanoparticles contain the chloroquine lipid, cationic lipid, phospholipid, PEG (polyethylene glycol) lipid and cholesterol. The chloroquine lipid can be used as a lipidated autophagy inhibitor, can endow the LNPs carrier with an autophagy inhibition function, and realizes the autophagy inhibition effect of'time-space synchronization ', so that the obstacle of'autophagy lysosome degradation' is broken through, and the gene delivery efficiency is enhanced.

The invention belongs to the technical field of biological medicine, discloses a novel venom polypeptide composite dendrimer as well as a preparation method and application thereof, and particularly discloses scorpion venom polypeptide, and the amino acid sequence of the scorpion venom polypeptide is FLGGSLSSIF. According to the invention, a novel scorpion venom polypeptide (marked as C9) is separated and identified, and a novel polypeptide-polyamidoamine dendritic compound G5C9 (i.e., nanoparticles) with a glioblastoma (GBM) targeting effect is constructed. Compared with the polypeptide C9, the cell uptake efficiency of the G5C9 is obviously improved. G5C9 can enter cells through endocytosis and target lysosome to destroy functions of the lysosome and mTORC1, so that nuclear translocation of TFEB is promoted, and autophagic death of the cells is induced. Therefore, the G5C9 provides a novel mode for targeting the anti-GBM in vitro and in vivo.

The invention discloses a nucleic acid tetrahedron nanostructure as well as a preparation method and application thereof. The nucleic acid tetrahedron nanostructure is composed of circRNA for coding an antigen, an ISD chain and two specific single-stranded DNA sequences, is simple and easy to prepare, can enter cells through cell membrane microfossa mediated endocytosis without the help of a transfection reagent, and is positioned to lysosome. The nanostructure is split to release circRNA coding antigen protein, and the circRNA coding antigen protein is delivered to APC and ISD chains in drainage lymph nodes (dLN) to activate cGAS-STING pathways, so that antigen presentation and immune activation are synergistically enhanced, T cells are efficiently and continuously activated, and the effect of tumor immunotherapy is achieved.

The invention provides regulatory elements, as well as vectors containing the same that may be used to stimulate transcription of a gene of interest in certain tissue types. The transcription regulatory elements described herein may be operably linked to a transgene, such as acid alpha-glucosidase (GAA), so as to promote expression of the GAA transgene in a cell, such as a muscle cell, liver cell, or neuron. The transcription regulatory elements described herein may be operably linked to a therapeutic transgene and used for the treatment of various disorders, such as lysosomal storage diseases, and particularly Pompe disease.

A cage-like nanocarrier for targeted delivery of siRNA, and a preparation method therefor and the use thereof. The preparation method comprises: (A) mutating a negatively charged or uncharged amino acid on the inner surface of a ferritin into a positively charged amino acid; and any one or more of the following steps: (B) coupling the N-terminus of the ferritin to a functional peptide having nucleic acid affinity; (C) coupling the N-terminus of the ferritin to a functional peptide promoting lysosomal escape; (D) truncating the E-helix at the C-terminus of the ferritin; (E) coupling the C-terminus of the ferritin to a functional peptide having nucleic acid affinity; and (F) coupling the C-terminus of the ferritin to a functional peptide promoting lysosomal escape. A new nucleic-acid-loaded protein nanocage carrier is constructed by means of modifying a negatively charged inner cavity of ferritin to make same positively charged. By means of electrostatic adsorption, a negatively charged siRNA can be efficiently loaded into a ferritin nanocage, thereby significantly improving the in-vivo and in-vitro delivery stability of siRNA, lysosomal escape functions, and efficacy of targeted therapy.

The invention belongs to the field of biological medicines, and discloses an application of a stem cell extract in preparation of a medicine for treating or preventing arthritis by regulating lysosome and / or mitochondrial function, a preparation method of the stem cell extract comprises the following steps: S1) culturing mesenchymal stem cells and producing stress conditions to stimulate the mesenchymal stem cells; s2) separating and purifying the culture supernatant of the mesenchymal stem cells cultured in the step S1) to obtain a stem cell extract; wherein S21) the culture supernatant is filtered to obtain a filtrate; the preparation method comprises the following steps of (S1) extracting a culture supernatant containing stress stem cell secretions through stress culture of stem cells, (S2) carrying out 3KD ultrafiltration concentration on the filtrate to obtain a crude sample, (S23) purifying the crude sample through molecular sieve exclusion chromatography or reversed-phase chromatography to obtain a purified sample, and (S24) taking the purified sample to form the preparation.

Provided herein are compositions and methods of using a bicistronic vector for treating or preventing a lysosomal storage disorder (LSD) in a subject. The disclosed compositions comprise a bicistronic vector comprising a promoter, an Internal Ribosome Entry Site (IRES), a polynucleotide encoding a lysosomal enzyme and a polynucleotide encoding a modified GlcNAc-1 phosphotransferase (GlcNAc-1 PTase). The present methods comprise administering to the subject a pharmaceutical composition comprising the bicistronic vector as disclosed herein.

The invention relates to a response type DNA tetrahedral lysosome targeting chimera as well as a construction method and application thereof, and belongs to the technical field of biology. The response type DNA tetrahedron lysosome targeting chimera is obtained by self-assembling a tetrahedron I and a tetrahedron II; a sticky tail end H1 is arranged on the tetrahedron I, and a sticky tail end H2 is arranged on the tetrahedron II; three tyrosine kinase receptor aptamers are anchored on the first tetrahedron, three lysosome shuttle receptor aptamers are anchored on the second tetrahedron, stimuli-responsive factor aptamers are hybridized on the cohesive end H2, and the binding capacity with cells is obviously enhanced based on a trivalent aptamer modification strategy of a tetrahedral framework. According to the present invention, the VEGF is adopted as the stimulation response factor to construct the responsive DNA tetrahedral lysosome targeting chimera, and the responsive DNA tetrahedral lysosome targeting chimera carries the HER2 protein to the lysosome under the mediation of the lysosome shuttle receptor IGF2R so as to degrade, such that the HER2 protein expression on the cell membrane is reduced, and the degradation specificity and the degradation effect are enhanced.