48 results about "Targeted killing" patented technology

The present invention provides proteins comprising immunoglobulin-type binding regions for cell-type specific targeting and Shiga toxin A Subunit effector regions for Shiga toxin effector functions (e.g. cellular internalization, directing subcellular routing, and / or cytotoxicity), wherein the binding regions and Shiga toxin effector regions are combined such that the Shiga toxin effector regions are proximal to the amino-terminals of the proteins. The presently disclosed proteins can comprise additional exogenous materials, such as, e.g., antigens, cytotoxic agents, and detection-promoting agents, and are capable of targeted delivery of these additional exogenous materials into the interiors of target cells. The proteins of the present invention have uses in methods such as, e.g., methods involving targeted killing of target cells, delivering exogenous materials into target cells, labeling subcellular compartments of target cells, and diagnosing and / or treating a variety of conditions including cancers, tumors, other growth abnormalities, immune disorders, and microbial infections.

The invention belongs to the pharmacological field of gene engineering, and provides a group of protein derivatives of human granzyme B by adopting gene engineering technology. The group of protein derivatives of the human granzyme B comprises GrB-G4S-GnRH (GrBLG) and mGrB-C4S-GnRH (mGrBLG) which are targeted fusion proteins formed by connecting human matured granzyme B (GrB) and mutagenic human matured granzyme B (mGrB) with human gonadotropin-releasing hormone (GnRH) through flexible connecting short peptide GlyGlyGlyGlySer (G4S) respectively and which can be combined with human GnRH receptors (GnRHR) on the surfaces of cells through ligand human GnRH. The mGrB eliminates the functions of the combination through a personal 'electrostatic exchange-absorption mode' and the entering into target cells of the prior GrB, and simultaneously reserves the enzymatic activity and the cytocidal function of the GrB. The invention provides experimental evidences for peculiarly targeted-killing GnRHR positive cells and minimizing toxic side effect by the fusion proteins, and shows the use of the group of protein derivatives of the human granzyme B in the targeted therapy on the positive adenocarcinoma of a type of gonadotropin-releasing hormone receptors.

The invention discloses a preparation method of cytotoxicity-enhanced efficient target killing NK / CIK (Natural Killer) / ( Cytokine Induced Killer) cells. The preparation method is applicable to culture of the NK / CIK and DC (Dendritic Cell)-NK / CIK cells derived from peripheral blood and cord blood. The preparation method comprises the following steps: acquiring blood; separating mononuclear cells; culturing NK / CIK cells by adopting suspending cells; culturing DC cells by adopting adherent mononuclear cells; carrying out mixed culture on the NK / CIK cells and the DC cells according to a proportion of 10: 1. According to the preparation method disclosed by the invention, mononuclear cells which are not sorted are adopted for culture for 14 days by utilizing efficient cytokine combination to generate NK / CIK mixed effector cells in a system, and the purity of the cells is nearly 90%, wherein NK cells account for 52.4%, and CIK cells account for 33.3%, and cells expressing killing marks account for over 80%, and all the NK cells highly express activating marks CD161; the induced DC-NK / CIK cells have remarkable multiplication capability and target killing capability to tumors, and the killing capability is remarkably improved especially in aiming at tumor cells insensitive to the NK cells, and thus the range of the NK cells in clinical tumor therapies is expanded.

The present invention provides protease-cleavage resistant molecules comprising Shiga toxin effector polypeptides capable of exhibiting potent, Shiga toxin functions (e.g. subcellular routing and cytotoxicity). The present invention also provides protease-cleavage resistant, cell-targeting molecules for targeting specific cell types, e.g., infected or malignant cells. Certain molecules of the present invention are cytotoxic, and certain cell-targeting molecules of the present invention may be used for the targeted killing of specific cell types and the treatment of a variety of diseases, disorders, and conditions, including cancers, tumors, growth abnormalities, immune disorders, and microbial infections. Certain cell-targeting molecules of the invention exhibit improved, in vivo tolerability as compared to related cell-targeted molecules comprising protease-cleavage sensitive, wild-type, Shiga toxin effector polypeptides. The cell-targeting molecules of the invention can deliver additional materials, such as, e.g., antigens, cytotoxic agents, and detection-promoting agents, into the interiors of target cells.

The invention discloses a conjugate, and a preparation method and application thereof. The conjugate has a structure of TP-L-D, wherein TP represents targeting peptide which is applicable to targeting the conjugate to tumor cells, and the surfaces of the tumor cells carrying lysosome tetraspanin LAPTM4B are used as a specific target of the targeting peptide; D represents a treatment activator; and L represents a linking arm which is used for connecting the treatment activator to the targeting peptide and connected with the treatment activator through a hydrazone bond. The conjugate provided by the invention can realize targeted killing of tumor cells carrying LAPTM4B protein on its surfaces, has strong targeting and destruction performance, is hard to incur immunoreactions, and can be used for treating or preventing tumors or preparing pharmaceutical compositions for treating or preventing tumors.

The invention discloses efficient targeting drug-loaded nano-micelle as well as a preparation method and application thereof. The preparation method comprises the following steps: (1) dissolving an amphiphilic block copolymer into an organic solvent, and then adding small molecule anti-leukemia drug hydrochloride; after the materials are fully dissolved and evenly mixed, adding triethylamine to remove hydrochloric acid so as to form a small molecule anti-leukemia drug; then, adding the small molecule anti-leukemia drug into a phosphate buffer solution (PBS) to form nano-micelle which has a hydrophilic shell and a core encapsulated with the drug; carrying out dialysis, and drying to obtain the drug-loaded nano-micelle; a polydopamine modification method is used for modifying a specific antibody against a leukemia cell surface antigen, thus realizing targeting. The drug-loaded nano-micelle prepared by the method has a particle size of 50-100nm, is very good in monodispersity, has a higher drug loading capacity and encapsulation efficiency, is simple in targeted modification step and good in targeted killing effect of leukemia cells, but is high in biosecurity for normal cells, and can realize slow release so as to avoid the defects caused by long-term and multiple dosing.

The invention relates to an antitumor stem cell antigen OCT4 (octamer-binding transcription factor 4) specific CTL (cytotoxic T lymphocyte) and a preparation method thereof. A directionally-targeted nano-liposome-carried tumor stem cell antigen is adopted to prepare the antigen-specific T lymphocyte. According to the tumor stem cell antigen-specific T lymphocyte preparation technique, the novel nano-liposome-carried tumor stem cell antigen is adopted, and the tumor stem cell is used as the therapeutical target, thereby inducing the more efficient targeted killing on the specific CTL of the tumor stem cell, and directly aiming at the tumor generation and relapse roots; and thus, the tumor therapy can implement a permanent cure. The 72-hour quick-maturated dendritic cell is adopted to greatly shorten the optimal therapeutical time window for the tumor patient. The engineering cell is adopted to amplify the antigen-specific T lymphocyte, thereby generating a high proportion of Tcm with higher killing capacity. The preparation method of the tumor stem cell antigen OCT4 specific T lymphocyte is simple and easy to implement; the immune target spot aims at the tumor stem cell; and thus, the antitumor stem cell antigen OCT4 specific CTL has high antigenicity and favorable stability.

A human telomerase reverse-transcriptase (hTERT) / human interleukin 18 (hIL 18) fusion protein with composite function is prepared from 2 cell factors with similar or complementary functions through artificial reforming of their linking terminals and configuring. The expression of its carrier is created for increasing its target killing power to tumor cells and the dendritic cell mediated immune-effect. It can be used to prepare the dendritic cell vaccine for treating cancer.

The invention relates to a SEA mutant gene and use for fusion protein thereof with ScFv gene, wherein gene engineering means is employed for saltation of super antigen SEA gene, which has the base sequence represented by the sequence table 1, by fusing it with anti-melanoma single-chain antibody (ScFv) gene obtained through bacteriophage displaying technique, a ScFv-SEA recombination antibody is constructed, wherein in vitro cell experiment proves that the fusion protein has inhibitory action to melanin oncocyte superior to other tumor, the fusion protein has evident targeting killing effect to melanin oncocyte.

The invention relates to the technical field of pharmaceutical biology. Flt3L is one of colony stimulating factor family members, and can be used for stimulating the proliferation and differentiation of CD34+hemopoietic stem cells. The invention aims to provide a nucleic acid vaccine adjuvant with the enhanced activity of cellular immunity and a construction method thereof. The nucleic acid vaccine adjuvant is a plasmid which is designed based on the Flt3L and is used for enhancing the cellular immune effect of a nucleic acid vaccine. In-vivo experiments prove that when simultaneously injected with the nucleic acid vaccine, the plasmid has the capacity of inducing a cellular immune reaction effectively, such as increasing of antigen-specific spleen IFN-gamma positive cells and enhancement of the target killing capacity of spleen CD8 positive cells. Therefore, the vaccine adjuvant of the invention can be added into the conventional nucleic acid vaccine preparation for mixing injection, so that the cellular immune reaction induced by the nucleic acid vaccine is promoted obviously, and the nucleic acid vaccine is used as a novel cellular immune adjuvant.

The invention belongs to the field of cell therapy, and especially relates to a chimeric antigen receptor modified lymphocyte capable of expressing CXCR4, and relates to genetic engineering means. According to the genetic engineering means, non-specific lymphocytes are modified into lymphocytes capable of identifying tumor surface related antigens and expressing chemotactic factor acceptor CXCR4,so that enrichment at specific parts is realized, and targeting killing on specific cells is realized. The invention also discloses a preparation method of the specific lymphocytes, and applications thereof in malignant tumor treatment.

The invention discloses a game information display method, a game information display device, a storage medium and an electronic device, wherein the method comprises the following steps of obtaining whole-field killing information in a game interface of a target game application, wherein the target game application is logged in through a target account, and the whole-field killing information includes target killing information relevant to the target account, and first killing information relevant to a virtual object different from a target virtual object controlled by the target account in the target game application; displaying the whole-field killing information on a first region of the game interface; and displaying the target killing information on a second region of the game interface, wherein the second region is different from the first region. The technical problems that in the prior art, the obtained killing information of the target virtual object operated and controlled bya game player in a game is not detailed enough, and the display position is not clear enough, so that the efficiency of obtaining the killing information of the target virtual object is low are further solved.

The present disclosure relates to compositions and methods for targeted killing of microorganisms. In particular, the present disclosure relates to the use of Lysobacter gummosus and compositions containing Lysobacter gummosus in targeted killing of microorganisms in medical, industrial, domestic, or environmental applications, as well as treatment of bacterial infections (e.g., in biofilms).

The invention relates to the technical field of cell culture, and discloses a high-efficiency proliferation reagent for peripheral blood NK (Natural killer) cells in vitro and an operation instruction. The reagent comprises the following components: an NK cell proliferation reagent: a gamma-ray inactivated recombinant K562 feeder cell, a CD52 antibody, an IL-2, a CD16 antibody, a CD3 antibody anda CD56 antibody; the gamma-ray inactivated recombinant K562 feeder cell is a K562 cell line for membrane expression of 41BBL and IL-15 which is lethally radiated by a 100G gamma-ray. According to thehigh-efficiency proliferation reagent for the peripheral blood NK cells in vitro and the operation instruction, the prepared NK cells have high purity and killing activity. Through detection, when theNK cells are cultured for 14 days, the purity of the NK cells can reach more than 90%; when the target ratio is 10:1, the killing activity of the NK cells on K562 cells can reach 80% or more; the proliferation speed of the prepared NK cells is high; when the NK cells are cultured for 14-17 days, the cell proliferation rate is more than 500; the NK cells prepared by the invention have excellent effects for targeted killing of tumor cells.

The invention relates to the field of walnut cultivation and planting, in particular to a diseases and insect pests prevention and treatment method for walnut. By adopting a mixed fertilization mannerof plant ash, potassium fertilizers and phosphate fertilizers, the fertility of walnut is satisfied, bacteria in the soil can be preliminary killed by plant ash, and soil structures can be also improved by plant ash, so that the mixed fertilization manner is relatively conducive to walnut tree growth. By preventive spraying of pesticides before germination, flowering and fruiting, targeted killing of germs and pests in the corresponding stages can be realized, and the disease incidence of walnut trees is reduced. Compared with a previous traditional post-treatment method that corresponding treatments are carried out after diseases or insect pests happen, the method has the advantages that pesticide expenditure cost is reduced, the fruiting rate of walnut trees is improved, and yield is increased.

The invention relates to a camptothecin prodrug gel, and a preparation method and use thereof, and belongs to the field of medicines. The present invention provides the camptothecin prodrug gel whichis prepared by polymerization of raw materials containing a camptothecin prodrug, methacrylic acid, a crosslinking agent and an initiator, wherein the structure of the camptothecin prodrug is as shownin a formula I. The PMAA-based pH / redox dual-responsive camptothecin prodrug gel provided by the invention has a particle size of a nanometer level, good pH response performance and glutathione response performance, and can rapidly release an active ingredient camptothecin in tumor cells to achieve the effect of targeted killing of the tumor cells. Biological experiments prove that the camptothecin prodrug gel of the invention can be successfully ingested by tumor cells, has significant inhibitory effect on tumor cells, exhibits obvious anti-tumor activity in vivo, and has a broad clinical application prospect.

The invention specifically relates to a recombinant vector for luciferase based on pCDH and application thereof, belonging to the field of genetic engineering. The recombinant vector is formed by tandem connection of a lentiviral vector pCDH and a luciferase gene and has a nucleotide sequence as shown in SEQ ID No. 1. A luciferase gene mediated by the recombinant vector expresses luciferase in raji cells; and in virtue of the principles of a reaction of a substrate under enzymatic catalysis, the recombinant vector is applied to tracking of CD19 in living imaging research on a model animal so as to evaluate the target killing effect of CAR-T cells.

The invention utilizes a genetic engineering technology for constructing an extracellular region LMP1 (latent membrane protein1) scFv segment of a chimeric antigen receptor targeting LMP1 with a high-affinity full human-derived anti-LMP1 Fab as a template. The extracellular region LMP1 scFv segment of LMP1 CAR is recombined with signal peptide CD8alpha, transmembrane region CD8TM, and intracellular regions CD28, CD137, and CD3zeta to construct a third-generation chimeric antigen receptor targeting LMP1. The prepared chimeric antigen receptor targeting LMP1 is recombined into a linearized lentiviral vector by overlap PCR, an anti-LMP1 CAR lentiviral recombinant expression vector is constructed, an LMP1 CAR virus solution is prepared, and T cells are infected to obtain LMP1 CAR-T cells. Thespecific targeted killing effect of the cells on LMP1 positive nasopharyngeal carcinoma cells is verified in vitro and in vivo, and new therapeutic ideas and means are provided for the treatment of nasopharyngeal carcinoma.

The invention provides an EBV (Epstein-Barr Virus) LMP2A (Latent Membrane Protein 2A) multi-epitope peptide for an immunological therapy and application of the EBV LMP2A multi-epitope peptide. The EBV LMP2A multi-epitope peptide for the immunological therapy is an amino acid sequence shown in SEQ ID NO.1. By adopting the technical scheme of the invention, after the EBV LMP2A multi-epitope peptide is loaded on DCs (Dendritic Cells) sourced from PB (Peripheral Blood) of a human body, the ripening ability and the presenting ability of the DCs can be more effectively promoted, and more cell factors can be secreted by the DCs; after the DCs on which the EBV LMP2A multi-epitope peptide is loaded are in co-cultivation with CTL (Cytotoxic T Lymphocytes), CTL cells in EBV specificity can be induced, an NPC (Nasopharyngeal Carcinoma) cell line CNE1 can be effectively subjected to targeted killing in vitro and in vivo, and the effect of the EBV LMP2A multi-epitope peptide is better than the effect of independently applying LMP2A356 to 364 or LMP2A426 to 434 or a mixture of the LMP2A356 to 364 and the LMP2A426 to 434; a better direction is provided for immune cell clinical therapy for treating NPC and other diseases related to EBV.

The invention relates to an active factor for enhancing lymphocyte targeting killing tumors and a preparation method thereof, wherein the amino acid sequence of the active factor is showed as SEQIDNO: 1. The preparation method comprises the following steps of: respectively amplifying the sections of SEQIDNO:2-4 by utilizing PCR (Polymerase Chain Reaction), inserting the three sections into a PQ31vector in series, and connecting flexible sections among all sections to obtain a fusion protein expression vector; transforming the fusion protein expression vector into an M15 escherichia coli strain, carrying out IPTG inductive expression, then washing an inclusion body, purifying by using Ni+-ITA resins under the condition of denaturation, and further renaturing to obtain a dsNKg2D-IL-15 protein. The preparation method can express a large number of dsNKg2D-il-15 proteins in vitro, and the expressed protein is stable and not degradable in physiological environment, and has the ability of ensuring that tumor cells are in anti-form presentation on IL-15 to activate the activities of NK cells.

The invention relates to a hygienic insecticide compound containing pirimiphos methyl and pyrethroid. Compared with a single agent, the compound has an obvious synergistic effect, can promote the insecticidal effect, can achieve a synergic coefficient above 130 and especially has excellent knock-down and killing effects to hygienic insects having pyrethroid resistance. Besides, after pirimiphos methyl with low toxicity, low stimulation and low use frequency is selected as a lethal agent of the hygienic insecticide compound and is compounded with ultrahigh knock-down efficient low-toxicity and low-stimulation pyrethroid chlorofluorothrin, d-trans-chloropropyrethrin, transfluthrin, sevoflurane methothrin or tetramethylfluthrin, the knock-down and lethality properties of the active ingredient to the target are both greatly promoted and the cost performance ratio is ultrahigh; furthermore, the compound is prepared into a micro-capsule suspension agent which has excellent physical and chemical properties and slow release effect, so that the target killing effect is greatly promoted; the preparation is low-toxicity and stimulation-free and is suitable for indoor and outdoor hygienic insect prevention and control.

The invention relates to novel use of annexin A3 (ANXA3) and particularly relates to an application of annexin A3 in preparation of a drug for target killing liver cancer stem cells. The invention puts forward the application of using ANXA3 as a liver cancer stem cell related antigen and then inducing to obtain the drug for target killing liver cancer stem cells by using liver cancer stem cell antigen-loaded ANXA3 for the first time. Anti-hepatoma specific immunotherapy based on the drug has a wide clinical application prospect.

The invention discloses a traditional Chinese medicine preparation for killing pyococcus. The medicine preparation is prepared from the following four kinds of ingredients in parts by weight: 20 to 45 parts of rhizoma polygoni cuspidate, 1 to 45 parts of euphorbia, 20 to 55 parts of Chinese violet and 40 to 60 parts of galangal through be ground into powder. Compared with the prior art, the traditional Chinese medicine preparation has the advantages that the injury of antibiotics on the human body is avoided; sterilization ingredients in the medicine preparation have the targeted killing effect on pyococcus.

The invention relates to the technical field of medical biology. Micro RNA-155 is micro ribonucleic acid (RNA) for regulating the maturation and natural immunoreaction of T cells and B cells. The invention provides a micro RNA-155-based plasmid for enhancing the immune effect of deoxyribonucleic acid (DNA) vaccine cells. A plasmid pcDNA 3.1-pre-miR-155 is obtained by inserting an expression sequence of a micro RNA-155 precursor molecule into a pcDNA-3.1 plasmid, and the micro RNA-155 precursor molecules can be expressed efficiently without influencing the expression capability of the co-transfected DNA vaccine. In an in-vivo test, the plasmid and the DNA vaccine are injected at the same time and have the capability of effectively introducing cellular immunoreaction, and such capability isrepresented by the increase of antigenically specific spleen IFN-gamma positive cells and enhancement of target killing capability of spleen CD8 positive cells. The plasmid of the invention can be added into the conventional DNA vaccine preparation and injected together so as to obviously promote the cellular immunoreaction induced by the DNA vaccine, and serves as a novel cell immune adjuvant.

The invention discloses a preparation method of a medicinal composition for killing aphids, which comprises the following steps: 1) preparing raw materials; 2) adding water to decoct the petunias, crane grass buds, and thunder pills for two times, and the medicinal liquid for later use; 3) Alisma, Sophora japonica rice, Thousand hammers, Nutmeg, Xu Changqing, Magnolia magnoli, decoct twice, filter, and concentrate the filtrate to a relative density of 1.15; 4) Merge the liquid medicine; 5) Adjust the pH value of the solution; 6) Filter , filled, sterilized, and obtained. The pharmaceutical composition described in the present invention can kill aphids in a targeted manner, has good safety, good biodegradation, no pollution and irritating odor, no environmental toxicity, no residue in animals, and does not destroy the ecological balance. The crop growth environment has little impact; it has good permeability and wetness, interferes with the central nervous system of insects, inhibits the synthesis and development of insect eggs, and is not easy to develop drug resistance; it has good quick effect and is suitable for widespread application in pest control.

The invention relates to the technical field of pharmaceutical biology. Flt3L is one of colony stimulating factor family members, and can be used for stimulating the proliferation and differentiation of CD34+hemopoietic stem cells. The invention aims to provide a nucleic acid vaccine adjuvant with the enhanced activity of cellular immunity and a construction method thereof. The nucleic acid vaccine adjuvant is a plasmid which is designed based on the Flt3L and is used for enhancing the cellular immune effect of a nucleic acid vaccine. In-vivo experiments prove that when simultaneously injected with the nucleic acid vaccine, the plasmid has the capacity of inducing a cellular immune reaction effectively, such as increasing of antigen-specific spleen IFN-gamma positive cells and enhancementof the target killing capacity of spleen CD8 positive cells. Therefore, the vaccine adjuvant of the invention can be added into the conventional nucleic acid vaccine preparation for mixing injection,so that the cellular immune reaction induced by the nucleic acid vaccine is promoted obviously, and the nucleic acid vaccine is used as a novel cellular immune adjuvant.