18 results about "Lysosomal membrane" patented technology

The invention belongs to the field of biomedicine, and discloses a method and a kit for monitoring a lysosome acidification process mediated by mitochondrial-lysosome interaction. According to the method, an LYSO-PZ probe with a specific structure is used for marking cell lysosome contents, a pH rodo dye and a lysosome membrane marker sfGFP-LAMP1-mCherry are combined for monitoring the pH value and membrane positioning of the lysosome respectively, then a three-channel fluorescence image is obtained through a structural illumination microscope (SIM), and the fluorescence intensity ratio is quantitatively analyzed. In addition, the method further comprises a step of forming mitochondrial-lysosome contact (MLC) through optogenetics regulation, related protein binding is induced through blue light, fluorescence intensity changes before and after MLC formation are compared, and MLC-mediated proton transport is verified. A complete monitoring system is established for the first time, lysosome acidification can be accurately monitored, information can be obtained in multiple dimensions, and the method has important clinical application value and wide market prospects.

The application belongs to the field of biomedicine, and discloses a method and a kit for monitoring a process of mitochondrion-lysosome interaction-mediated lysosome acidification. The method is characterized in that a cell lysosome content is first labeled with a LYSO-PZ probe with a specific structure, a pHrodo dye and a lysosome membrane marker sfGFP-LAMP1-mCherry are combined to monitor the pH value and membrane localization of the lysosome, respectively, then three-channel fluorescence images are acquired through a structured illumination microscope (SIM), and the fluorescence intensity ratio is quantitatively analyzed. In addition, the method further comprises a step of optogenetic regulation of mitochondrion-lysosome contact (MLC) formation, the fluorescence intensity change before and after the formation of MLC is compared through blue light induction of related protein binding, and the proton transport mediated by MLC is verified. The application first establishes a complete monitoring system, can accurately monitor lysosome acidification, and has important clinical application value and broad market prospect.

The application discloses a low-oxygen and pH dual-response nano-carrier, a drug-loaded nanoparticle and application thereof, and belongs to the technical field of biological medicines. The nano-carrier with moderate particle size and low-oxygen and pH dual-response is obtained by using the reaction of aldehyde benzoic acid, nonaethylene glycol and azo-p-phenetidine, the nano-carrier is specifically targeted to a tumor site, and then specifically releases drugs, thereby reducing toxic side effects; meanwhile, after the nano-carrier responds, benzaldehyde groups are released, the benzaldehyde groups are covalently connected with proteins on a lysosome membrane, the structure and activity of the proteins are changed, the lysosome membrane is broken after the proteins are denatured, the permeability of the lysosome membrane is increased, the nano-carrier cannot be discharged to the extracellular, the drug concentration in cells is greatly improved, and the drug bioavailability of the nano-carrier is significantly improved; in addition, the drug loading capacity of the nano-carrier is high, and therefore, the nano-carrier has a good application prospect.

To provide a new pharmaceutical composition for treating tumors.SOLUTION: In order to treat and / or prevent a tumor in a subject, a lysosomal membrane permeability enhancer is used in combination with the following a) and / or b): A) administration of an autophagy inhibitor; b) intake restriction of lysine or administration of a lysine antagonist.SELECTED DRAWING: None

The invention relates to lysosome-based nano drug-loaded particles and application thereof, and belongs to the technical field of biological medicines. According to the invention, the lysosome membrane is used as a biomimetic membrane source to coat the surface of the drug nanoparticle, so that the structure stability of the lysosome membrane is maintained, and the activity of hydrolase which naturally exists is retained to the greatest extent, so that the drug-loaded nanoparticle can perform in-situ degradation on an extracellular matrix of a tumor tissue in an in-vivo delivery process; therefore, the physical barrier of the tumor tissue is effectively weakened, the permeation, distribution and cellular uptake ability of the nano-drug in the tumor tissue are remarkably enhanced, the condition that immune cells enter the tumor tissue is improved on the basis, and finally the tumor treatment effect is improved.

The application provides a tertiary amine lipid compound with a structure shown in formula (I) or a stereoisomer or a pharmaceutically acceptable salt thereof, and an application thereof. The tertiary amine lipid compound is hydrophobic and electrically neutral at normal physiological pH, can be self-assembled into pH-responsive membrane-breaking material nanoparticles with a PEG shell layer together with materials such as DSPE-PEG, has weak interaction with membranes and is low in cytotoxicity; under a slightly acidic pH condition, part of the tertiary amine can be protonated to form an amphiphilic structure composed of a hydrophobic domain and a cationic domain, has strong interaction with cell membranes and organelle membranes and strong membrane-breaking activity, and thus can efficiently and selectively destroy cell membranes or lysosome membranes, so as to kill tumor cells.

The invention relates to a pH response type amphiphilic xanthophyll polypeptide nano-drug as well as a preparation method and application thereof. Xanthatin is efficiently modified by polypeptide with a side chain containing a urea acylhydrazide group, the polypeptide is encapsulated into a nano carrier to obtain the nano preparation, a cRGD targeting element on the side chain enhances the targeting property of the drug to breast cancer cells, the drug enters the breast cancer cells through a lysosome way, a urea acylhydrazone chemical bond is broken to form urea acylhydrazide in an acid environment, and the drug has a good anti-tumor effect on breast cancer cells. The pH-responsive release of the xanthophyll raw medicine is realized, meanwhile, urea hydrazide forms and induces lysosome membrane rupture, tumor cell apoptosis is caused through a mitochondrial pathway, and lysosome escape of the xanthophyll raw medicine is promoted, so that ROS is enhanced and cell apoptosis is promoted, and effective treatment of breast cancer is realized through the synergistic effect of urea hydrazide and the xanthophyll raw medicine.

The present invention discloses a fluoroboron dipyrrole cyclometallated iridium (III) complex, a synthesis method thereof, and applications thereof. The complex is obtained by connecting fluoroboron dipyrrole with cyclometallated iridium to obtain a photosensitizer with high reactive oxygen species generation capacity, high anticancer activity, and low toxic side effects. The complex has good cellular uptake capacity and the ability to target lysosomes in tumor cells. It can generate reactive oxygen species through near-infrared light irradiation, causing lysosomal membrane permeability, the release of acidic hydrolases leading to tumor cell acidification, and calcium ion homeostasis imbalance causing mitochondrial damage, which is specifically manifested as a decrease in membrane potential and intracellular ATP level, inducing tumor cell apoptosis and autophagy blockade, and further inducing immunogenic cell death, thereby achieving good anti-tumor activity and having broad application scenarios.

The application discloses an HSC70 protein K128R mutant and application thereof in regulating CMA activity. The application discloses application of ubiquitination modification of an HSC70 protein at a K128 site in regulating chaperone-mediated autophagy. It is found for the first time that the HSC70 protein is subjected to K11-connected polyubiquitination under metabolic stress, and the modification site is a lysine at the 128th position (K128). The modification is mediated by a ROS-ATM-Atg7-TRIM21 signal axis, and is a necessary condition for the combination of the HSC70 and a lysosome membrane receptor LAMP2A to be enhanced and CMA to be activated. By constructing a K128R mutant, it is proved that the modification loss can block the response of CMA to metabolic stress. The application provides a new diagnostic marker and a drug target for CMA-related diseases (such as tumors and neurodegenerative diseases), and has important clinical application value.

The invention relates to the technical field of biological medicine, in particular to application of glucosylceramide level as a diagnosis and treatment target of heart failure. According to the invention, key pathological cascade reactions such as lysosomal membrane permeabilization, cathepsin leakage, mitochondrial dysfunction and the like in the occurrence and development process of heart failure are blocked from the source by reducing the level of glucosylceramide in myocardial tissues. By inhibiting synthesis of glucosylceramide or enhancing degradation in lysosome of glucosylceramide, glucosylceramide abnormally accumulated in myocardial cells is removed, organelle structures and energy metabolism states are remarkably improved, and then myocardial contraction and relaxation functions are improved. In an animal model, the strategy can reverse ventricular remodeling, improve ejection fraction and relieve cardiac hypertrophy and fibrosis. In addition, the glucosylceramide detection method provided by the invention can be used for assisting in evaluating metabolic abnormalities related to heart failure, so that disease judgment is more accurate.

The present invention relates to a composition comprising an anti-CD20 antibody and a PIKfyve inhibitor for preventing or treating CD20 positive cancer such as B cell lymphoma. The present invention can maximize the anticancer effect of an anti-CD20 antibody, specifically, obinutuzumab, by co-administering an anti-CD20 antibody and a PIKfyve inhibitor, which is a negative regulator of a tumor cell killing mechanism thereof. In addition, by using PIKfyve as a screening target, the present invention can rapidly identify, with high reliability, combination therapy candidate agents that can synergistically enhance the direct cell death (DCD), lysosomal membrane permeabilization (LMP), and subsequent cancer cell-killing activity of anti-CD20 antibodies.

The present application relates to a kind of lysosome-based nano drug-loaded particles and its application, belong to the field of biological medicine.The present application will be lysosome membrane as the source of biomimetic membrane to be coated on the surface of drug nano-particle, while keeping the stability of lysosome membrane structure Maximum degree retains its naturally occurring hydrolytic enzyme activity, so that the nano drug-loaded particle can be in situ degradation to tumor tissue extracellular matrix during in vivo delivery, thereby effectively weakening the physical barrier of tumor tissue, significantly enhance the permeability, distribution and cell uptake capacity of nano drug in tumor tissue, and on this basis, improve the conditions for immune cells to enter tumor tissue, ultimately improve the tumor treatment effect.

The invention discloses a screening method of a traditional Chinese medicine monomer composition for resisting heart failure. The method comprises the following steps: 1, carrying out water extraction and enrichment on astragalus membranaceus and monkshood to obtain astragalus membranaceus and monkshood soup enriched components; 2, synthesizing myocardial mitochondrial membrane coated magnetic beads and myocardial lysosome membrane coated magnetic beads; 3, gathering enriched components of the astragalus mongholicus-monkshood decoction, and carrying out ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry to obtain components of songorine, neoline, calycosin and formononetin as traditional Chinese medicine monomer composition; the traditional Chinese medicine monomer composition is prepared by dissolving songlin, neoline, calycosin and formononetin in a sodium carboxymethyl cellulose solution. The synthesized myocardial mitochondrial membrane coated magnetic beads and the myocardial lysosome membrane coated magnetic beads are used for adsorbing and identifying active ingredients in the astragalus mongholicus-monkshood decoction, the effective ingredient composition for resisting heart failure is successfully screened, and the preparation method is simple and easy to implement; the traditional Chinese medicine monomer composition plays a role in preparing anti-heart-failure medicines by improving heart functions and the like.

The application belongs to the technical field of biological materials, and particularly relates to an enzyme response in-situ self-assembled peptide and anti-tumor application thereof. An amino acid sequence of the self-assembled peptide PSK is shown in SEQ ID NO. 1. The self-assembled peptide PSK of the application firstly combines a PD-L1 blocking peptide with a tumor cell surface PD-L1, effectively internalizes the PSK peptide into the tumor cell through intracellular endocytosis, hydrolyzes in a lysosome through Legumain response bond, triggers self-assembled module assembly to form a beta-folded nanofilament structure, further triggers lysosome membrane permeability change, escapes from the lysosome, releases KLA to destroy mitochondria and induce tumor cell apoptosis. At the same time, the PD-L1 binding peptide is endocytosed into the lysosome, releases immunosuppression, combines with ICD induced by mitochondrial damage, enhances the killing effect of immune cells on tumor cells in the tumor microenvironment, effectively improves the selectivity and anti-tumor efficacy of the polypeptide, and has good biological safety.

The invention belongs to the technical field of biological medicine, and relates to a tandem amplification pyroptosis nano inducer, a preparation method thereof and application of the tandem amplification pyroptosis nano inducer in preparation of cancer treatment drugs. The tandem amplification pyroptosis nano inducer is of a nano-particle structure and comprises crystalline dendritic Gd2O3 NPs and a shell packaged on the surface of the crystalline dendritic Gd2O3 NPs, crystalline dendritic Gd2O3 nano-particles load CPPO and Ce6, and the shell is an inorganic layer formed by calcium carbonate nano-particles. The tandem amplification pyroptosis nano inducer can promote NF-kappa B pre-activation through pH response and replace a phosphate group on a lysosome membrane so as to cause lysosome rupture; in addition, the released CPPO and Ce6 generate singlet oxygen, so that the limitation of light penetration and tumor hypoxia is overcome, strong anti-tumor immune response can be initiated in vitro and in vivo through the synergistic effect with pyroptosis, and the tumor immune curative effect is further improved.

Lysosomal membrane permeabilization is an evolutionarily conserved hallmark of stress-induced cell death. Here the inventors show that the major stress-inducible heat shock protein 70 (Hsp70) enhances cell survival by stabilizing lysosomes through a pH-dependent high affinity binding to an endo-lysosomal anionic phospholipid bis(monoacylglycero)phosphate (BMP; also referred to as lysobisphosphatidic acid). The positively charged ATPase domain of Hsp70 is responsible for the binding but the substrate-binding domain is also required for effective stabilization of lysosomes. Importantly, the cytoprotective effect can be obtained by endocytic delivery of recombinant Hsp70 and specifically reverted by extra cellular administration of BMP antibodies or Hsp70 inhibitors. Thus, this protein-lipid interaction opens exciting possibilities for the development of cytoprotective and cytotoxic lysosome-specific therapies for the treatment of degenerative diseases and cancer, respectively.

The invention discloses an intelligent drug co-delivery liposome for targeted therapy of Alzheimer's disease as well as a preparation method and application of the intelligent drug co-delivery liposome. The invention relates to a brain-targeted peptide-responsive liposome, which is composed of a brain-targeted peptide-grafted maleeimine-polyethylene glycol-phosphatidylethanolamine (DSPE-PEG2000-ANG2), an ROS (reactive oxygen species) responsive lipid DSPE-TK-PEG2000, hydrogenated soybean phospholipid (HSPC), cholesterol (Chol), and a GSK-3beta inhibitor 7BIO and dipalmitoyl phosphatidylserine (DPPS), wherein the GSK-3beta inhibitor 7BIO and the dipalmitoyl phosphatidylserine (DPPS) are used as drugs. According to the present invention, the intracerebral delivery efficiency of 7BIO is significantly improved, the acidification disorder of lysosome in damaged neurons is improved while the damaged lysosome membrane is recovered, the aggregation deposition of A beta is inhibited, the systemic toxicity is reduced, and the safe and efficient new strategy is provided for the treatment of Alzheimer's disease.

The invention discloses an aggregation-induced emission (AIE) polymer probe with lysosome targeting viscosity response as well as a preparation method and application of the AIE polymer probe. According to the probe, tetraphenyl ethylene (TPE) is used as an AIE luminous skeleton, and two hydrophilic flexible side chains are introduced by utilizing the excellent structural modifiability of the tetraphenyl ethylene (TPE). The tail end of each side chain is modified with pyridine positive ions, and the pyridine positive ions can specifically target a lysosome membrane through electrostatic interaction; the three ether groups embedded in the side chain effectively enhance the water solubility of the molecule. According to the invention, TPE and an amphiphilic flexible side chain are used to construct an AIE polymer, a TPE-derived AIE polymer probe (PTPEB-Py) is designed, and a drug action time-lysosome viscosity-fluorescence lifetime (t-eta-tau) three-dimensional linear relationship is constructed through intracellular and extracellular fluorescence lifetime signals so as to visualize a lysosome viscosity change value. Therefore, quantitative detection and visualization of lysosome viscosity change in the mitochondrial autophagy process in cancer cells are realized.