32 results about "Granzyme B" patented technology

Abstract Described herein, in one aspect, are antigen presenting cells (APCs) comprising an exogenous nucleic acid encoding one or more candidate antigens, wherein the one or more candidate antigens are expressed and presented with MHC class I or MC class II molecules; a molecular reporter of Granzyme B (GzB) activity; and c) an exogenous inhibitor of caspase-activated deoxyribonuclease (CAD)-mediated DNA degradation, a CAD knockout, or a caspase knockout (e.g., caspase 3 knockout). Described herein, in another aspect, is a system for detection of recognized antigen presentation by an antigen presenting cell to a cytotoxic lymphocyte or NK cell. Abstract 2018 / 22761 oM - cell Target ml Target cell cell cell Target Target Target SUBSTITUTE SHEET (RULE 26) cell cell cell Target Target cell cell 1 / 28 my Isolate recognized cell Library of target cells target cells and displaying different Add T cells from sample sequence antigens antigens of interest. CTLs deliver cytotoxic granules to target cells displaying cognate antigen FIG. 1 PCT / US2018 / 036663 20 26 20 53 56 07 J ul 2 02 6 2 0 2 6 2 0 5 3 5 6 0 7 J u l 2 0 2 6 2 0 1 8 / 2 2 7 6 1 o M a n d m y 1 / 2 8 m y L i b r a r y o f t a r g e t c e l l s d i s p l a y i n g d i f f e r e n tA d d T c e l l s f r o m s a m p l e of interest. CTLs deliver c y t o t o x i c g r a n u l e s t o t a r g e t c e l l s d i s p l a y i n g c o g n a t e a n t i g e n P C T / U S 2 0 1 8 / 0 3 6 6 6 3

The invention discloses shRNA (short hairpin Ribonucleic Acid) targeting Zeb2, a recombinant vector, a cell and application of the shRNA in preparation of drugs for enhancing anti-tumor immune response, and belongs to the technical field of research and development of anti-tumor drugs. Experiments prove that the shRNA targeting Zeb2 can reduce the proportion of T cells (Tex, PD1 + Tim3 +), increase the proportion of depleted precursor T cells (Tpex, Ly108 + Tim3-), improve the secretion ability of Granzyme B and TNF-alpha cytokines of CD8 + T cells in tumor tissues, significantly inhibit the tumor growth of skin melanoma mice, increase the infiltration quantity of CD8 + T lymphocytes in the tumor tissues and the anti-tumor ability of the CD8 + T cells, and can be used for preparing anti-tumor drugs for treating skin melanoma. The polypeptide has the potential of being used for preparing drugs for enhancing anti-tumor immune response.

The invention discloses application of an ATP (adenosine triphosphate) citrate lyase (ACLY) inhibitor in preparation of a medicine for treating leucoderma, and belongs to the technical field of preparation of medicines for treating leucoderma. Through targeted inhibition of ACLY activity or specific knockout of ACLY in CD8 + T cells, abnormal activation of ACLY and secretion of killing effector molecules (Granzyme B, IFN-gamma, Perforin and the like) are inhibited, and infiltration of the CD8 + T cells to skin lesion parts is reduced, so that melanocytes are protected from immune injury, and the progress of vitiligo is blocked. The application is based on the abnormally high expression of ACLY in CD8 + T cells of vitiligo patients and the close association with disease activity, and a brand new targeted treatment thought and a medicine research and development direction are provided for vitiligo.

The invention relates to the technical field of biological medicine, in particular to an immune synergist, an enhanced NK cell preparation, a preparation method and application. The immune synergist is hirudin, and the concentration of the hirudin is 10-40 [mu] g / ml. The enhanced NK cell preparation is prepared by the preparation method. According to the preparation method, the enhanced NK cell preparation is obtained after NK cells which are amplified in vitro for 14 days are pretreated by adopting the immune synergist. The application comprises an application of the immune synergist in up-regulation of expression of NK cell granzyme B and an application of the immune synergist in preparation of an NK cell preparation for inhibiting breast cancer cell activity. The expression level of granzyme B of the NK cells can be safely and economically improved, and the in-vitro killing activity of the NK cells to breast cancer cells is remarkably enhanced.

The application provides application of psoralen in activating NK cells. The psoralen provided by the application is a coumarin component extracted from traditional Chinese medicine psoralea; through in-vitro cell experiment verification, the psoralen can activate NK-92 to secrete IFN-gamma, Granzyme B and Perforion, and activate surface protein NKG2D of NK-92; meanwhile, the psoralen also has the same effect on a cell group containing NK cells; through in-vivo experiment verification, the psoralen helps to improve the total amount of NK cells in the mouse spleen, inhibit tumor growth, and improve the expression amount of IFN-gamma, Granzyme B, Perforion, NKG2D, NKP46 and CD107a in a tumor infiltration environment. In summary, the small-molecule compound psoralen has good effect in activating NK cells.

The application belongs to the technical field of biological medical materials, and particularly relates to a granzyme B specific response nano probe and a preparation method and application thereof. The granzyme B specific response nano probe comprises a near-infrared down-conversion nano probe, and a pentapeptide IEFDK-fluorescence quenching modification group is modified on the surface of the near-infrared down-conversion nano probe; the pentapeptide IEFDK-fluorescence quenching modification group is derived from a pentapeptide IEFDK-fluorescence quencher with a structure shown in formula (1). The granzyme B specific response nano probe provided by the application can specifically respond to granzyme B, realize immune effect monitoring of a tumor site, and thus realize detection of tumor immune responses after different radiotherapy schemes are processed. Formula (1).

The present application provides a dendrobium officinale polysaccharide extract, a preparation method and an application, the dendrobium officinale polysaccharide extract is with dendrobium officinale stem as raw material, is extracted by purified water reflux, is precipitated by ethanol, is deproteinized by Sevag, is decolorized and is deashed and is obtained by gel / film separation and purification, the mass percentage content of polysaccharide in the extract is 50-95wt%, the average molecular weight is 10-300 kDa, and the monosaccharide component includes mannose, glucose, galactose, fucose, rhamnose and arabinose.The dendrobium officinale polysaccharide extract provided by the present application has immune activation effect on natural killer cells, can promote the NK cell CD107a degranulation reaction and the expression of IFN-gamma, perforin and granzyme B in a dose-dependent manner, up-regulates the NKG2D related pathway, thereby activating the NK cell function and enhancing the killing activity of NK cells on tumor target cells such as K562.

Compounds of Formula (I) and Formula (II), which are capable of binding to granzyme B. Also provided herein are pharmaceutical compositions comprising such for use in, for example, imaging Granzyme B and / or treating immunoregulatory abnormalities.

The invention relates to an application of beta-damascenone (beta-damascenone) in preparation of a medicine for inhibiting CD8 < + > T cell depletion. The invention discovers that beta-damascenone can significantly down-regulate the expression of a depletion marker TIM3 of CD8 + T cells and maintain the secretion level of Granzyme B and TNF-alpha, so that the cytotoxic function of the T cells is not damaged while the depletion of the T cells is inhibited. The use includes the use of beta-damascenone as a single active ingredient or in combination with an immune checkpoint inhibitor (such as an anti-PD-1 antibody) for the treatment of cancer, such as melanoma, non-small cell lung cancer or colorectal cancer. The invention further provides an in-vitro method for enhancing the anti-tumor function of the CD8 + T cells.

Compositions and methods are provided for the cell-mediated targeted killing of diseased cells based on the presence of an intracellular antigen, rather than a surface-bound marker. The targeting cells are modified to express a cytotoxic protein that is delivered into a targeted cell, and after delivery is selectively activated by the presence of a cytoplasmic protein of interest. In one embodiment of the invention, the cytotoxic molecule is a Granzyme B (GrB) polypeptide. In the compositions of the invention, GrB is modified to render its cytotoxic enzymatic functions inactive, until the presence of an intracellular antigen unlocks the GrB molecule to enable enzymatic activities.

Novel cyclic peptides that bind to granzyme B and may be suitable for imaging granzyme B, their salts, pharmaceutical compositions containing them, diagnostic and therapeutic uses, and methods for producing such compounds are disclosed. Additionally, compounds useful as radiotracers for positron emission tomography (PET) and / or single photon emission computed tomography (SPECT) imaging are provided. The use of the compounds as imaging agents for granzyme B is further disclosed.

This invention relates to a granzyme B-targeting complex, a radiopharmaceutical, and their imaging applications in tumor immunotherapy, belonging to the field of nuclear medicine diagnostics. The complex is covalently modified, triggering a covalent group proximity effect by recognizing the binding of a peptide to granzyme B, achieving irreversible covalent linkage between the probe and the target protein. This significantly enhances the probe's selective retention at the target site and extends the imaging time window. After radionuclide labeling, the complex can be prepared into a radiopharmaceutical for nuclear medicine imaging; the labeling process is simple and exhibits good stability. This radiopharmaceutical allows for non-invasive, specific, and quantitative monitoring of granzyme B expression levels via PET or SPECT, reflecting the activity status of immune cells in vivo. This invention has significant clinical application value and promising prospects for the early and accurate prediction and dynamic monitoring of tumor immunotherapy efficacy.

The invention provides a dendrobium officinale polysaccharide extract as well as a preparation method and application thereof.The dendrobium officinale polysaccharide extract is prepared from dendrobium officinale stems as a raw material through purified water reflux extraction, ethanol precipitation, Sevag deproteinization, decoloration and deliming and gel / membrane separation and purification, the mass percentage content of polysaccharide in the extract is 50-95 wt%, the average molecular weight is 10-300 kDa, and the content of polysaccharide in the extract is 50-95%. The monosaccharide components comprise mannose, glucose, galactose, fucose, rhamnose and arabinose. The dendrobium officinale polysaccharide extract provided by the invention has an immune activation effect on natural killer cells, can dose-dependently promote the degranulation reaction of NK cells CD107a and the expression of IFN-gamma, perforin and granzyme B, and up-regulate NKG2D related pathways, so that the functions of the NK cells are activated, and the killing activity of the NK cells on tumor target cells such as K562 is enhanced.

The invention discloses an application of Danjie emodin in preparation of a product for inhibiting T cell depletion. It is found through in-vitro screening that the Danjie emodin can significantly inhibit T cell depletion. Mechanism research shows that the Danjie emodin does not influence the secretion of Granzyme B and slightly inhibits the secretion level of a cell factor IFN-gamma; by inhibiting an AKT phosphorylation signal channel, accumulation of protein aggregates in T cells is remarkably reduced, and the cell protein homeostasis is maintained, so that T cell depletion is inhibited. Animal experiments also prove that the T cells treated by the Danjie emodin can significantly inhibit the growth of melanoma after adoptive reinfusion. Therefore, the invention provides the application of the Danjie emodin in preparation of the product for inhibiting T cell depletion, and further provides a new small molecule metabolism intervention strategy for improving the solid tumor immunotherapy effect.

The application provides an application of dodartin A1 in activating NK cells. Through in-vitro experiments, it is proved that dodartin A1 can significantly promote the synthesis and release of IFN-gamma and cytotoxic substances (such as granzyme B and perforin) of NK92 cells, and increase the expression of the surface agonistic receptors NKp46 and NKG2D of the NK92 cells; through in-vivo experiments, it is proved that dodartin A1 can promote the expression of the NK cell activation related substances CD107a and NKp46 to play an anti-hepatoma effect; in conclusion, dodartin A1 has the effect of activating NK cells.

The invention provides application of B cell granzyme B as a target spot in preparation of a product for treating myocardial infarction or heart failure, and belongs to the technical field of biomedicine. The invention provides the effect of inhibiting the expression of B cell granzyme B and treating myocardial infarction or heart failure for the first time. Animal experiments show that by inhibiting the expression of B cell granzyme B, the cardiac function after myocardial infarction can be improved, the expression of heart failure indexes after myocardial infarction can be reduced, the expression of inflammatory factors in the heart after myocardial infarction can be inhibited, and cardiac fibrosis caused by myocardial infarction can be improved. The application has the advantages that the myocardial infarction or the heart failure can be improved by inhibiting the B cell-derived granzyme B for the first time, a new prevention and treatment drug research and development approach and a drug action target are provided for the treatment of the myocardial infarction and / or the heart failure, and the application value is very important.

The application provides a biomarker for diagnosing GVHD and application thereof. The application proves by experiments that the Ki67 positive proportion, Granzyme B positive proportion and TEM cell proportion in donor-derived T cells can be used as biomarkers for distinguishing GVHD patients from non-GVHD patients, and when the T cells cannot be distinguished from the donor and the recipient, the Ki67 positive proportion, Granzyme B positive proportion and TEM cell proportion in the T cells can still be used as biomarkers for distinguishing GVHD patients from non-GVHD patients. The method for diagnosing GVHD by using the biomarker provided by the application has the advantages of immune subpopulation specific quantification, low cell amount adaptability, short time efficiency, low cost and the like.

The invention relates to the technical field of tumor immunotherapy, in particular to application of an AKR1B1 inhibitor and an immune checkpoint inhibitor in preparation of antitumor drugs and a pharmaceutical composition containing the AKR1B1 inhibitor and ICIs. After the AKR1B1 inhibitor is combined with the immune checkpoint inhibitor, the following technical effects are achieved: 1, the tumor growth is obviously inhibited: in a mouse breast cancer model and a lung cancer model, the tumor growth inhibition rate of the epalrestat single drug is about 30%; the inhibition rate of a PD-1 single drug is about 30%-40%; the inhibition rate of combined treatment is remarkably improved to 65%, and an obvious synergistic anti-tumor effect is shown. 2, the anti-tumor immune response is enhanced, and the activity of CD8 positive T cells can be remarkably improved by the epalrestat single drug; the combined treatment can improve the proportion of cytokines (such as granzyme B and interferon g) secreted by CD8 positive T cells in tumors. 3, the safety is high: under the administration dosage, no obvious drug toxicity is observed in the heart, liver, spleen, kidney and lung of the mouse;

Compounds of Formula (I) and Formula (II) that are capable of binding to granzyme B. Also provided herein are pharmaceutical compositions comprising the compounds, for example, for imaging and / or treating immunomodulatory abnormalities of granzyme B.

The application discloses a fluorescence hybridization chain reaction biosensor based on the existence of granzyme B and application thereof, and realizes real-time detection of T cell effector molecules by establishing a fluorescence hybridization chain reaction biosensor based on the existence of active granzyme B, and has the characteristics of short amplification time, simple operation, high amplification efficiency, simple reaction system and high economic benefit. By introducing peptide nucleic acid into the hybridization chain reaction system, the cutting of the substrate by granzyme B is successfully used as the starting recognition event of the hybridization chain reaction, and after the cutting of the peptide substrate in the peptide nucleic acid by granzyme B, the foothold is exposed, the strand displacement reaction is mediated, the fluorescence signal is restored after amplification, and a new strategy of using the hybridization chain reaction for detecting active cytokines is provided. The strategy makes the hybridization chain reaction using fluorescence as a signal output mode applicable to active protein detection, and has the advantages of a simple reaction system, few operation steps and low detection cost.

The invention relates to a method for activating NK (Natural Killer) cells and application thereof, in particular to application of an LINC02577 inhibitor in preparation of an NK cell activating medicine. Experiments show that the expression quantity of the LINC02577 is reduced in the NK cell activation process, so that the LINC02577 can be used for preparing a kit for detecting NK cell activation; meanwhile, the knock-down LINC02577 is found to be capable of effectively promoting expression of NK cells CD107a and granzyme B, promoting secretion of IFN-gamma and TNF-alpha and promoting the killing activity of the NK cells on leukemia cells, so that the knock-down LINC02577 can be used for preparing the medicine for promoting activation of the NK cells, and the NK cells with the knock-down LINC02577 can be used for preparing the medicine for treating leukemia.

The invention discloses a fusion polypeptide, an anti-KIR2DL1 monoclonal antibody and an anti-aging composition containing the fusion polypeptide and the anti-KIR2DL1 monoclonal antibody. Wherein the amino acid sequence of the fusion polypeptide is SEQ ID NO: 1. A heavy chain variable region of the anti-KIR2DL1 monoclonal antibody is SEQ ID NO: 2, and a light chain variable region of the anti-KIR2DL1 monoclonal antibody is SEQ ID NO: 3. The composition composed of the two components regulates NK cells bidirectionally by enhancing an NKG2D activation pathway and blocking a KIR2DL1 inhibition pathway, so that the proportion of CD107a positive NK cells reaches 68.9%, the secretion amount of granzyme B reaches 465pg / mL, the survival rate of senescent cells is reduced to 39%, and the synergistic effect exceeds 89%. The composition can be prepared into intravenous injection drugs, oral health care products or external cosmetics, is safe and remarkable in synergistic effect, and solves the problems of insufficient curative effect of a single activator and poor specificity of a single inhibitor in the prior art.

Compounds that bind to granzyme B and can contain a radioactive moiety, such as the compound of formula (I), and pharmaceutical compositions comprising the same. Uses of the compound and pharmaceutical composition in cancer treatment, as well as kits and methods for cancer treatment, are also provided herein.

The invention discloses a Rhodol fluorescent dye, an enzyme response type fluorescent probe and composition thereof, and a preparation method and application of the Rhodol fluorescent dye and the enzyme response type fluorescent probe and composition. The Rhodol fluorescent dye is shown as a formula (I), a formula (II), a formula (III), a formula (IV), a formula (V) or a formula (VI), or the formula (I), the formula (II), the formula (III), the formula (IV) or the formula (VI). Wherein the dye of the formula VI obtains stable fluorescence performance through ortho-position carboxyl methyl esterification. Based on the dye shown in the formula VI, two enzyme response type fluorescent probes are further provided and are respectively used for detecting the activity of caspase-3 and the activity of granzyme B. The two enzyme response type fluorescent probes are combined to obtain the protease double-target fluorescent probe composition which can synchronously monitor upstream and downstream key signals in an immune killing pathway. The series of materials have important application value in noninvasive curative effect monitoring and evaluation of tumor immunotherapy.

The invention belongs to the technical field of biomedical materials, and particularly relates to a granzyme B specific response nanoprobe as well as a preparation method and application thereof. The granzyme B specific response nanoprobe comprises a near-infrared down-conversion nanoprobe, and the surface of the near-infrared down-conversion nanoprobe is modified with a pentapeptide IEFDK-fluorescence quenching modification group; the pentapeptide IEFDK-fluorescence quenching modification group is derived from a pentapeptide IEFDK-fluorescence quenching agent with a structure as shown in a formula (1). The granzyme B specific response nanoprobe provided by the invention can specifically respond to granzyme B, and realizes monitoring of immune effect of a tumor site, so that detection of tumor immune response after treatment of different radiotherapy schemes is realized. Formula (1)

A kind of granzyme B targeting complex, radiopharmaceutical and its imaging application in intestinal immune abnormal activity belong to the field of nuclear medicine diagnosis and treatment.The complex is covalently modified, specifically non-covalently combined with granzyme B by recognizing peptide first, and then triggers irreversible covalent connection by means of proximity effect, effectively solves the technical bottleneck of low target tissue uptake, short in vivo retention time and narrow imaging window of existing granzyme B probe, significantly improves the selective retention capacity and imaging specificity of the probe in the target site.The complex and radiopharmaceutical of the present application can specifically target granzyme B released by intestinal lesion site immune abnormal activity, realize non-invasive, dynamic, quantitative nuclear medical imaging of intestinal inflammation such as Crohn's disease, ulcerative colitis and aGVHD, and accurately evaluate the degree of intestinal immune abnormal activity and monitor the treatment effect, providing a core tool for early diagnosis and precise diagnosis and treatment of such diseases, and having important clinical transformation value and application prospect.

This invention discloses the application of emodin in the preparation of products that inhibit T cell exhaustion. In vitro screening revealed that emodin significantly inhibits T cell exhaustion. Mechanistic studies showed that emodin does not affect Granzyme B secretion, slightly inhibits the secretion level of the cytokine IFN-γ, and significantly reduces the accumulation of protein aggregates within T cells by inhibiting the AKT phosphorylation signaling pathway, thus maintaining cellular protein homeostasis and inhibiting T cell exhaustion. Animal experiments also confirmed that T cells treated with emodin significantly inhibited melanoma growth after adoptive infusion. Therefore, this invention provides the application of emodin in the preparation of products that inhibit T cell exhaustion, further offering a new small-molecule metabolic intervention strategy for improving the efficacy of immunotherapy for solid tumors.

The present application relates to the technical field of NK cell culture, and discloses a method for improving the killing function of NK cells and application thereof.The method comprises the following steps: (1) adding a haem oxygenase 1 antibody to an NK cell culture medium to prepare an antibody culture medium; (2) performing sterile filtration treatment on the antibody culture medium; and (3) separating NK cells and culturing the NK cells in the antibody culture medium.The present application adds a haem oxygenase 1 antibody to treat NK cells, blocks the expression of haem oxygenase 1 in the NK cells, or inhibits the activity of an effective substance of haem oxygenase 1.After being treated with the haem oxygenase 1 antibody, the secretion of granzyme B and perforin of the NK cells can be promoted, and the killing effect of the NK cells on target cells Yac-1 and K562 can be improved, so that the anti-tumor function of the NK cells is improved.

The invention provides a marker composition, a kit and a prediction and evaluation method for evaluating the risk of immune reconstitution insufficiency of an HIV (Human Immunodeficiency Virus) infected person, and the marker composition, the kit and the prediction and evaluation method are used for predicting and evaluating the risk of immune reconstitution insufficiency of the HIV infected person by adopting the combination of three markers of an interferon gamma-induced mononuclear cell chemokine (MIG), a nerve growth factor (NGF) and granzyme B (GZMB). Effective evaluation and auxiliary diagnosis of the risk of incomplete immune reconstitution (INR) of an HIV infected person under long-term anti-retrovirus treatment are realized, the prediction sensitivity, specificity and stability are improved, and a basis is provided for clinical early intervention and individualized management.