99 results about "Lysosomal targeting" patented technology

The present invention provides highly phosphorylated lysosomal hydrolases, methods of modifying lysosomal hydrolases with the lysosomal targeting pathway enzymes GlcNAc-phosphotransferase and / or phosphodiester alpha-GlcNAcase.

The present invention provides nucleotide and amino sequences of the lysosomal targeting pathway enzyme phosphodiester alpha-GlcNAcase, methods of producing and methods of purifying this enzyme.

The invention discloses a lysosome-targeted fluorescent dye capable of realizing red emission and near-infrared emission, and a preparation method and application thereof, belonging to the field of bioluminescence analysis. The preparation method comprises the following steps: dissolving a compound with the substituent R in an anhydrous organic solvent and adding morpholinoindolal and a catalyst under the condition of introduction of nitrogen, wherein a mol ratio of morpholinoindolal to the compound with the substituent R is 1-10: 1; and carrying out a reaction at a reaction temperature of 25 to 200 DEG C for 1 to 24 h, concentrating an obtained solution and carrying out silica-gel column chromatography so as to obtain the target fluorescent dye. The fluorescent dye prepared in the invention is applicable to targeted imaging of lysosomes in cells, fluorescent probes or laser dyes. The fluorescent dye has the advantages that the emission wavelength of the fluorescent dye is in a range from red zone to near-infrared zone; the fluorescent dye can prevent interference of biological background fluorescence and has high fluorescence quantum efficiency and good light stability; and the fluorescent dye can be specifically localized in lysosomes, so the fluorescent dye has high application value.

The invention discloses a lysosome targeted fluorescence probe for detecting hypochlorous acid molecules, as well as a preparation method and application thereof, and belongs to the technical field of analytical chemistry. The molecular formula of the fluorescence probe is C37H35N5O5S, with a structure represented by formula I shown in the description. The synthesis of the probe includes four steps, and the after-treatment process is relatively simple; the selective quick detection of the hypochlorous acid molecule probe is achieved, the selectivity is good, and the capability of resisting interference from other molecules is strong; in addition, the change in color of the solution can be observed with the naked eyes, and the change in fluorescence color can also be observed under an ultra-violet lamp, therefore, the fluorescence probe for detecting hypochlorous acid molecules is one with the chromogenic sensing function, can be applied to detection of hypochlorous acid in lysosome in the cells of the biological system, and can be used as a probe for detecting hypochlorous acid in lysosome in the cells. The lysosome targeted fluorescence probe for detecting hypochlorous acid molecules, provided by the invention, is a simple, quick and sensitive reagent for detecting the hypochlorous acid molecules, and has a wide application prospect in the field of biomolecule detection.

The invention relates to a lysosome targeted fluorescent probe as shown in a structural formula and used for lactam ring leuco body regulation and control. The fluorescent probe responds to weakly acidic environment (with pH of 4-6) in a highly sensitive and highly selective manner in a complex buffer solution through lactam ring OFF-ON allosterism. Fluorescent confocal imaging experiments of the probe show that the probe can recognize lysosomes in tumor cells in a targeted manner and has certain promising application prospect in the diagnosing of diseases such as cancers.

The invention provides a lysosome hypochlorous acid fluorescence probe which has a chemical name of 8,8,9-trimethyl-5-(2-morpholinoethyl)-9,10-dihydrobenzo[de] pyrrole[2,3-9]isoquinoline-4,6(5H,8H) diketone, called TPS-Lyso-HClO for short. The lysosome hypochlorous acid fluorescence probe is capable of detecting the content of hypochlorous acid in solution and lysosome: the excitation wavelength of the solution is detected to be 380nm; when the pH value is 7.0-7.4, the emission wavelength is 446nm; when the pH value is 5-5.5, the emission wavelength is 490nm; the two-photon cell fluorescence imaging excitation wavelength is 780nm, and the emission bands refer to 480-540nm and 560-620nm; and the single photon cell fluorescence imaging excitation wavelength is 405nm, and the emission bands refer to 480-540nm and 560-620nm. The lysosome hypochlorous acid fluorescence probe disclosed by the invention is high in specificity of a hypochloric acid reaction and is capable of resisting against multiple interferents; and the fluorescence probe has lysosome targeting property and is high in sensitivity, low in detection limit and wide in detected hypochloric acid concentration range. The lysosome hypochlorous acid fluorescence probe disclosed by the invention is a simple, rapid and sensitive hypochloric acid molecular specificity detection reagent and has wide application prospects in the field of bio-molecular detection.

The invention provides a photosensitive lysosome targeted hypochloric acid fluorescent probe and a synthesis method and application thereof to detection of aqueous solution and lysosome hypochloric acid. The fluorescent probe takes fluorescein as fluorophore, takes 2-nitrobenzyl bromide as a photosensitive protecting group and takes morpholine as a lysosome targeted positioning group. The lysosome targeted hypochloric acid fluorescent probe is used for sensing detection of hypochloric acid in aqueous solution or biological systems, and sensing detection includes fluorescence detection and cell imaging. Ultraviolet irradiation is required before sample detection, the excitation wavelength is 480nm in solution detection and is 488nm in cell detection, and fluorescence peaks appear at positions of 525nm. The lysosome targeted hypochloric acid fluorescent probe is simple in synthesis step, and product purification is simple; high specificity in hypochloric acid reaction and resistance to various interferences are realized; due to introduction of the lysosome positioning group, a targeting effect is achieved, fluorescence reaction is controllable temporally and spatially, and the photosensitive lysosome targeted hypochloric acid fluorescent probe has a promising application prospect in the field of biomolecule detection.

The invention relates to a fluorescent probe which can directly detect hypochlorite in lysosome on the basis of 1,8-naphthalimide and a preparation method and application of the fluorescent probe. The fluorescent probe has the molecular structure as shown in (I), is simple to synthesize, can selectively detect hypochlorite ions in a pure PB buffer solution and can resist interference from other active oxygen small molecules. Besides, by use of the fluorescent probe, color changes of the solution can be observed with naked eyes and also can be observed under an ultraviolet lamp. The fluorescent probe can also be used for detecting hypochlorite ions in the cytolysosome, is short in response time and low in limit of detection and has a potential application value.

The invention belongs to the technical field of analysis and detection, and provides a lysosome-targeted Cys near-infrared fluorescent probe and a preparation method and application thereof. The near-infrared fluorescent probe is 3-(2-(3-(dicyanomethylene)-5,5-dimethylcyclohexen-1-yl)vinyl)-4-morpholinyl coumarin-7-acrylate which is abbreviated as DCICA. The prepared lysosome-targeted Cys near-infrared fluorescent probe has the characteristics of novel structure, lysosome targeting performance and near-infrared fluorescence emission (wherein an emission wavelength is 680 nm), can realize high-selectivity high-sensitivity detection of a Cys content of a solution, conducts specific detection of Cys in cell lysosome by using a laser confocal imaging technology, is of important guiding significance to research on the molecular mechanism of participation of the Cys in the life activity of the lysosome, and has a wide application prospect.

The invention discloses a benzothiazole lysosome-targeted pH fluorescent probe and preparation and application thereof. A preparation method of the probe comprises the steps that quinoline-4-formaldehyde, 2-methyl-benzothiazole and trimethylchlorosilane (TMSCl, serving as a catalyst) are dissolved into dimethylformamide according to the mole ratio of 1:1:10 in a sealed tube, the solution is heated to 100 DEG C, a reaction is conducted for 16 hours, and precipitates are filtered out; the precipitates are dissolved with dichloromethane, the pH value is regulated to be 8.0 with a NaCO3 solution, extraction is conducted with dichloromethane, drying and recrystallizing are conducted, and the pure product is obtained. The probe has the large Stokes displacement (110 nm) and has the high sensitivity and selectivity on H<+> changes. The pKa value is 3.52, and the pH linear range is 3.0-3.8. A laser co-focus experiment shows that the probe can be positioned to a lysosome in a targeted mode and makes a response for pH changes of the lysosome in a weak acid environment. In addition, the probe can detect pH changes in escherichia coli in a highly acidic environment (the pH value is smaller than 4) in a highly sensitive mode.

The invention relates to an IR780 double-targeted near-infrared fluorescent probe preparation method and application on tumor diagnosis and treatment, wherein the compound comprises two molecular units including an IR780 mother nucleus and morpholine. In the same molecular structure, mitochondria targeting, lysosome targeting, near-infrared fluorescence imaging diagnosis and photodynamic treatmenton tumors are integrated. The invention also discloses the application on tumor diagnosis and treatment. The fluorescent probe has the advantages of a simple structure, small molecular weight, a determined chemical structure, easy preparation and purification processes, low toxicity to mice and the like, so that the fluorescent probe can meet the basic requirements of clinical medication. In-vitro experiments prove that by means of the near-infrared fluorescence imaging, the electropositivity of tertiary amine and the negative electronegativity of the mitochondria in the near-infrared fluorescence probe structure based on IR780 tumor double targeting can obviously target tumor cells and tumor tissues, can generate photodynamic treatment action under near-infrared radiation, and can obviously inhibit tumor growth.

The invention discloses a lysosomal targeting fluorescent probe and a preparation method thereof. A D-Pi-A-Pi-D (donor-Pi-receptor-Pi-donor)-type large Pi conjugated system is constructed by alpha, beta-unsaturated ketone as a receptor. Through intramolecular charge transfer, compared with the existing many fluorescent probes for lysosome, lysosomal targeting fluorescent probe can realize large red shift of the excitation wavelength and emission wavelength of the molecule produce so that background signal interference is effectively reduced. The compound provided by the invention is used for fluorescence imaging of lysosome. In the cell environment, red fluorescence can be emitted under acidic conditions, and the maximum emission wavelength is about 625nm so that the lysosome targeting marker is obtained and background interference is small. The preparation method is sensitive, simple and quick.

The present invention provides methods for treating Pompe disease in a subject by administering to the subject a therapeutically effective amount of a fusion protein which includes human acid alpha-glucosidase (GAA), or a fragment thereof, and a lysosomal targeting domain. The lysosomal targeting domain binds the human cation-independent mannose-6-phosphate receptor in a mannose-6-phosphate-independent manner.

The invention discloses a lysosome-targeted hypochlorous acid near-infrared fluorescent probe as well as a preparation method and application thereof, which belongs to the technical field of analysischemistry. The lysosome-targeted hypochlorous acid near-infrared fluorescent probe is characterized in that a structural formula of the lysosome-targeted hypochlorous acid near-infrared fluorescent probe is as follows: (shown in the description). The invention also discloses a preparation method of the lysosome-targeted hypochlorous acid near-infrared fluorescent probe as well as application in water environment or biological cell system detection. The fluorescent probe of the invention has the characteristics of near-infrared transmission, high sensitivity, good selectivity, rapidness in response and the like.

Antibodies are disclosed that bind to and inhibit the anti-coagulant function of TFPI and have a lower affinity for TFPI at pH 6.0 than at pH 7.4. The lower affinity at pH 6 improves circulating half-life (T½) due to reduced target mediated clearance, a process by which an antibody / antigen complex is endocytosed and trafficked to the lysosome where both components are degraded. The lower affinity at pH 6.0 results in disruption of the complex prior to lysosome targeting and allows for re-circulation of the antibody. Specific modifications to antibody binding by histidine residue substitution are disclosed along with methods of use.

The invention relates to the technical field of pH fluorescent probes and particularly relates to a lysosome targeted pH fluorescent probe for monitoring cell autophagy as well as preparation and application of the lysosome targeted pH fluorescent probe. The preparation method comprises the following steps of dissolving 2-(2-aminoethyl)-3',6'-bis(diethylamino) spiro [isoindole-1,9'-xanthan]-3-ketone and 2-(2-methoxyethoxy)4-methyl benzene sulfonic acid ethyl ester in N, N-dimethylformamide, and carrying out heating reflux to obtain a crude product; and removing a solvent from the crude product, and separating through a silica gel column to obtain a pure product. Cytotoxicity tests show that the probe has almost no toxic or side effect on cells, a cell co-localization experiment determinesthat the probe can specifically target a cell lysosome, and a laser confocal microimaging experiment shows that the probe has good cell membrane permeability and can perform high-sensitivity detectionon pH change in the lysosome. The probe provided by the invention can monitor the autophagy process of the cells by detecting the change of pH in the lysosome.

The invention discloses a lysosome targeting hypochlorous acid ratio fluorescent probe and its preparation method and application. The molecular formula of the probe is: C 49 h 55 N 7 o 5 , the structural formula is as follows: The probe adds a lysosomal positioning group at the reaction site, and utilizes the energy transfer between two fluorescent groups to cause a change in fluorescence. A ratiometric probe for recognizing hypochlorous acid is designed and the response time is very fast, which can be applied Sensing detection of hypochlorous acid in a biological cell system; the sensing detection includes fluorescence detection, visual qualitative detection, and cell imaging detection.

The invention provides a lysosome-targeted photo-controlled fluorescent molecular switch, and a synthesis method and an application thereof. The specific molecular structure of the molecular switch is 3-amino-substituted rhodamine ethylmorpholinylamide, and the structural formula of the molecular switch is represented by formula (1). The lysosome-targeted photo-controlled fluorescent molecular switch of the invention has acid resistance, and also retains the photoactivation performance, so like acid-resistant photoactivated dyes can be used in a super-resolution imaging technique in lysosomes without being interfered by the pH value of a biological environment in order to achieve a good imaging effect. In addition, the lysosome-targeted photo-controlled fluorescent molecular switch of the invention can also be used in the fields of sensing and detection as a molecular fluorescent probe.

The invention provides an application of fluorescent carbon dots based on N-methyl o-phenylenediamine hydrochloride in lysosome target. The fluorescent carbon dots are prepared by dissolving the N-methyl o-phenylenediamine hydrochloride into absolute ethyl alcohol; the fluorescent carbon dots can quickly penetrate through cytomembrane and be gathered in lysosome, to be applied to washing-free lysosomal imaging of living cell and living bodies. The carbon dots in the invention are very simple to synthesize and convenient to operate, has fluorophores, and have the advantages of good light stability and high quantum yield; the fluorescent carbon dots can quickly enter cells, and can be naturally targeted at the lysosome; and the fluorescent carbon dots can be applied to quick and washing-freeimage in the living cells and living bodies.

The invention belongs to the technical field of nano probes, and discloses a double-organelle-targeted nano probe as well as a preparation method and application thereof. The double-organelle-targetednano probe is mainly prepared from a compound of formula I and a photo-sensitive agent II; and the photo-sensitive agent II is phthalocyanines or porphyrins photo-sensitive agent with negative charges. The compound of formula I is a mitochondria-targeted chemotherapy reagent with an aggregation-induced luminescent effect. The nano probe of the invention not only can monitor the release process ina cell in real time by virtue of a double-fluorescent illumination way, but also can effectively improve the killing efficiency for cancer cells by virtue of the synergism of the mitochondria-targeted chemotherapy and lysosome-targeted photodynamics therapy. The invention also relates to application of nano probe in preparing an antitumor drug and / or fluorescent imaging. (The formulas are shown in the description).

The invention provides a fluorescent probe for detecting biological mercaptan in lysosome. The chemical structural formula of the fluorescent probe is shown in the specification. According to the probe, in the presence of Cys and Hcy, the probe generates two different fluorescence emissions of blue and green at 480nm and 550nm respectively under the excitation of two independent wavelengths. However, when GSH and Na2S are added into the probe, blue fluorescence can only be generated at 480 nm. The difference can be reasonably attributed to the fact that the NBD-GSH / SH intermediate is differentfrom the NBD Cys / Hcy, and an intramolecular cyclization rearrangement reaction cannot occur. Meanwhile, due to the fact that morpholine has lysosome targeting, lysosome can be distinguished from other organelles. The fluorescent probe has the advantages of good lysosome targeting, strong specificity, fast response and the like and can be used for real-time visual determination of biological mercaptan in living cell lysosome, and can be obtained through chemical synthesis and has the advantages of simple and feasible synthesis process, cheap and easily available raw materials, low preparationcost and easiness in popularization.

The invention relates to a lysosome targeted fluorescent substance of which the structural formula is shown in the specifications. The synthesis method of the compound comprises the following steps: taking trifluoroacetic acid as a solvent and a reaction catalyst, enabling 4-bromaniline and paraformaldehyde to react to generate a bromine-substituted intermediate product, and then, carrying out Heck reaction to substitute bromine by 4-vinyl-N,N-dimethyl benzylamine. The method of the invention effectively applies the Heck reaction and has simple synthesis paths and higher yield, and the obtained product has stable properties and has the functions of fluorescence emission of visible light and lysosome targeting.

The invention provides a lysosome targeting fluorescent probe and a synthesis method and cell imaging application thereof. The fluorescent probe takes naphthalimide as a parent, an azacyclo-ketone substituent is introduced to the 4 site, an N-(2-aminoethyl) morpholine substituent is introduced to the N site, lysosome can be rapidly and accurately marked, the fluorescent probe is shown in the formula (1), R is the azacyclo-ketone substituent, and azacyclo-rings comprise a quaternary azacyclo-ring, a five-membered azacyclo-ring, a six-membered azacyclo-ring and the like. The excitation energy required by fluorescence emission of the probe is relatively low, and the probe can be used for labeling lysosome in living cells and performing two-photon fluorescence imaging at low concentration. And the fluorescent probe can penetrate deep tissues in two-photon fluorescence imaging, so that the probe can become a powerful tool for studying life processes such as lysosome transportation.

The invention relates to synthesis and an application of a leave-on near-infrared lysosome probe, belonging to the technical field of biology. A synthetic method of a malachite green lysosome targeted fluorescent probe comprises the following steps: synthesizing substituted aldehyde by adopting p-hydroxy benzaldehyde and ethyl 4-bromobutyrate in acetone by taking potassium carbonate as an acid-binding agent, synthesizing ethyl butyrate modified malachite by adopting the substituted aldehyde and N,N-dimethylaniline in ethanol by taking ZnC12 as a catalyst, hydrolyzing the malachite in acetone by taking a potassium hydroxide solution as a catalyst, and oxidizing butyrate derivate malachite in ethyl acetate by taking 2,3-dichloro-5,6-dicyan p-benzoquinone as an oxidizing agent, so as to obtain the malachite green lysosome targeted fluorescent probe. Intracellular experiments prove that the malachite green gyro type near-infrared targeted lysosome probe has good membrane permeability and can be specifically activated by lysosome; no fluorescence exists outside the lysosome; and therefore, leave-on lysosome marking purposes are achieved.

The invention discloses a naphthalimide-rhodamine-based compound. The compound is represented by general formula (I) shown in the description, and all substituent groups in the formula (I) are as defined in the description. The invention also discloses a detection composition and a detection kit of the naphthalimide-rhodamine-based compound, and a use of the detection composition or the detectionkit in the detection of mercury ions in a water environment and a biological cell system as a fluorescent probe for lysosomal targeted detection of mercury ions. The compound realizes the rapid detection of mercury ions by the fluorescent probe, has a good selectivity, and is a fluorescent probe having a chromogenic sensing function.

The invention discloses an organelle targeting photosensitizer capable of activating pyroptosis of tumor cells and a preparation method and application thereof. The Mito-ZS photosensitizer is a mitochondrial targeting photosensitizer; the Lyso-ZS photosensitizer is a lysosome targeting photosensitizer; the ER-ZS photosensitizer is an endoplasmic reticulum targeting photosensitizer. The photosensitizer activates a pyroptosis pathway by performing precise targeting therapy on organelles in tumor cells so that photodynamic therapy and immunotherapy strategies are perfectly combined, and the photosensitizer has a wide application prospect in the field of cancer therapy.

The invention belongs to the technical field of bioluminescence detection materials, and particularly relates to lysosome-targeted infrared two-window emission fluorescent dye as well as a preparationmethod and application thereof. According to the invention, an aniline derivative and a five-membered heterocyclic conjugated derivative are connected to an aza-fluoroborane parent structure, so thatthe infrared two-window emission of the material is realized. The target fluorescent dye has good light stability, thermal stability and chemical stability; excellent photo-thermal performance, photoacoustic imaging and tumor photo-thermal treatment properties are also shown; meanwhile, the preparation method of the lysosome-targeted infrared two-window emission fluorescent dye provided by the invention has theoretical basis support for design and synthesis, and has the advantages of simple preparation method, mild synthesis conditions, abundant raw materials and high target product yield, thereby having important guiding significance for designing and synthesizing efficient diagnosis and treatment reagents.

The invention discloses a lysosome targeted two-photon viscosityfluorescence probe and a preparation method and application thereof. The viscosityfluorescence probe uses naphthalimide as a parent, and the structure of the probe is shown in the description. The viscosityfluorescence probe can respond to fluorescence signals having specificity to viscosity. A cytotoxicity test indicates that the lysosome targeted two-photon viscosityfluorescence probe has almost no toxic and side effect on cells, a confocal fluorescence microimaging experiment indicates that the probe has good cell permeability to MCF-7 cells, a lysosome co-positioning experiment indicates that the probe has the good positioning effect on cell lysosome, and the probe can be suitable for detection of the viscosity in the cell lysosome.

The invention provides a lysosome target fluorescent probe of sulfur dioxide / sulfurous acid (hydrogen) saline which can be used for qualitatively or quantitatively detecting the content of sulfur dioxide or derivatives in solutions, tissues or zebra fishes. The lysosome target fluorescent probe of sulfur dioxide is simple in synthesis step, raw materials are easy to obtain, the yield is high, andthe lysosome target fluorescent probe of sulfur dioxide is suitable for industrialized application; and meanwhile, the identification speed is high, interferences of various kinds of ions, amino acidand reactive oxygen can be resisted, and the lysosome target fluorescent probe of sulfur dioxide has the relatively good specificity, can be used for detecting variation of sulfur dioxide in cells andzebra fishes, and has potential application value for research of biological pathology and relevant diseases.