82 results about "P53 proteins" patented technology

There is provided a novel compound that inhibits interaction between murine double minute 2 (Mdm2) protein and p53 protein and exhibits anti-tumor activity. The present invention provides an imidazothiazole derivative represented by the following formula (1) having various substituents that inhibits interaction between Mdm2 protein and p53 protein and exhibits anti-tumor activity:wherein R1, R2, R3, R4, and R5 in the formula (1) each has the same meaning as defined in the specification.

Specific sequences in the human genome are the sites of strong binding of wild-type p53 protein, but not mutant forms of the protein. These sequences are used diagnostically to detect cells in which the amount of wild-type p53 is diminished. The sequences can also be used to screen for agents which correct for loss of wild-type p53 to DNA in cancer cells.

The invention is in the field of cancer treatment. In particular, the present invention provides pharmaceutical compounds capable of interacting with mutant and non-mutant forms of cancer-related regulatory proteins such that the mutant protein regains the capacitv to properly interact with other macromolecules thereby restoring or stabilizing all or a portion of its wild type activity. Regulatory proteins include members of the p53 protein family such as. for example, p53, p63 and p73. The compounds of the invention are useful for cancer treatment. Methods for screening for such pharmacological compounds are also provided.

Systemic lupus erythematosus (SLE) can be prevented or treated by down-regulating the autoimmune response to the C-terminal-DNA-binding domain of the p53 protein (p53) by an active principle selected from the group consisting of: (i) a peptide of, or comprising, the C-terminal DNA-binding domain of the p53 protein; (ii) a monoclonal antibody (mAb) specific for said domain of p53 (Ab1), and fragments thereof; (iii) an mAb specific for Ab1 (hereinafter Ab2), and fragments thereof; (iv) a peptide based on a complementarity determining region (CDR) of the heavy or light chain of said Ab1 or Ab2; (v) a DNA molecule coding for (i) and (iv) of for the variable region of said Ab1 and Ab2 of (ii) and (iii); and (vi) T cells specific for (i) to (iv), fragments thereof, T cell receptor (TCR) thereof and peptides comprising the variable region of said TCR. SLE can also be diagnosed by assaying for antibodies (Ab1) against the C-terminal DNA-binding domain of p53 or antibodies (Ab2) specific to the Ab1 antibodies.

In fibrotic lung fibroblasts, basal levels of p53 protein (and miR-34a) are markedly suppressed, leading to reduced p53-mediated inhibition of uPA and uPAR, or concurrent induction of PAI-1. These changes contribute to excessive FL-fibroblast proliferation and production of extracellular matrix (ECM), and, therefore, pulmonary fibrosis. These processes are reversed by treating the cells, and treating subjects suffering from idiopathic pulmonary fibrosis (IPF) with the small organic molecule nutlin-3a (NTL) or with a peptide, CSP-4 (SEQ ID NO:1), or variants or derivatives or multimers of this peptide, which increase p53 levels by inhibiting MDM2-mediated degradation of p53 protein. Use of these compounds serves as a new approach to the treatment of IPF, they restore p53 expression and p53-mediated changes in the uPA-fibrinolytic system in FL-fibroblasts and restrict production and deposition of ECM.

Provided herein are peptidomimetic macrocycles containing amino acid sequences with at least two modified amino acids that form an intramolecular cross-link that can help to stabilize a secondary structure of the amino acid sequence. Suitable sequences for stabilization include those with homology to the p53 protein. These sequences can bind to the MDM2 and / or MDMX proteins. Also provided herein are methods of using such macrocycles for the treatment of diseases and disorders, such as cancers or other disorders characterized by a low level or low activity of a p53 protein or high level of activity of a MDM2 and / or MDMX protein.

The present invention relates to a novel use of Msx1 protein or a nucleotide encoding the same for inducing apoptosis. The Msx1 of the present invention induces apoptosis through direct interaction with p53 via a homeodomain and such interaction leads to increased stability, and / or nuclear localization of p53 in cells. The Msx1 or homeodomain thereof can be effectively used for the treatment of tumors, in which wild-type p53 protein has lost its function by some mechanism that inactivates p53 proteins.

The invention relates to a fusion protein polymer. The fusion protein polymer consists of a component A and a component B, wherein the component A is a single domain antibody, while the component B is peptide sequences which perform multimerization on fusion protein and are derived from TNF alpha protein, ACRP30 protein, P53 protein, COMP protein, C4BP protein and mutants of the TNF alpha protein, the ACRP30 protein, the P53 protein, the COMP protein and the C4BP protein. The fusion protein polymer has the advantages of successful expression in Escherichia coli, high expression quantity, relatively simple preparation and low cost. Poly-antibodies of the fusion protein polymer have high dissolubility and biological activity and greatly improve the affinity of antibody protein. The fusion protein polymer can be prepared into various biotech products such as diagnostic reagents used for diagnosing melanoma and the like, and used for target treatment of the melanoma and the like by carrying a therapeutic medicament. When the poly-antibodies based on the fusion protein polymer are clinically applied, the problem of strong immunogenicity can be solved.

The invention relates to an esophageal squamous cell carcinoma autoantibody molecular marker model and an application thereof. The molecular model mainly comprises an ALDOA autoantibody, an ENO1 autoantibody, a p53 autoantibody, and an NY-ESO-1 autoantibody, and can be used for preparing a kit for distinguishing esophageal squamous carcinoma patients and healthy medical examiners. The kit for detecting esophageal squamous cell carcinoma patients mainly comprises recombinant ALDOAD protein, recombinant ENO1 protein, recombinant p53 protein, and recombinant NY-ESO-1 protein. According to the esophageal squamous cell carcinoma autoantibody molecular marker model and the application thereof, the ENO1 autoantibody is found to be elevated in serum level in the esophageal squamous carcinoma patients for the first time, and is jointly detected with the ALDOA autoantibody, the p53 autoantibody, and the NY-ESO-1 autoantibody for distinguishing the esophageal squamous cell carcinoma patients andthe healthy medical examiners, and has a better distinguishing effect than a single index detection; and in addition, the detection method adopted by the invention is an enzyme-linked immunosorbent assay indirect method, is simple and convenient to implement, has good sensitivity and specificity, and is a method which is mature and reliable and can be widely used in base layer hospitals.

Isolated polypeptides, isolated polynucleotides or expression vectors encoding same, viral display vehicles which can be specifically bind an exposed epitope shared by mutant, but not wild type, p53 protein are provided. Also provided are methods of inducing apoptosis and treating cancer as well as diagnosing a p53-related cancer using the isolated polypeptides uncovered by the present invention.

There is provided compounds that inhibit interaction between murine double minute 2 (Mdm2) protein and p53 protein and exhibits anti-tumor activity. Compounds include imidazothiazole derivatives that can inhibit interaction between Mdm2 protein and p53 protein and exhibits anti-tumor activity.

The present invention provides novel compounds, corresponding to formulae I and II, respectively, which are able to reactivate the apoptosis-inducing function of mutant p53 proteins. This reactivation is provided by restoration of sequence-specific DNA-binding activity and transcriptional transactivation function to mutant p53 proteins, and modulation of the conformation-dependent epitopes of the p53 protein. Accordingly, the substances according to the invention will be used in pharmaceutical compositions and methods for treatment of patients suffering from various types of tumors.

The invention discloses a high-sensitivity protein detection method, wherein one of the following modes is selected optionally: mode 1, performing a tube wall solid phase PLA (proximity ligation assay) by a direct coating method, namely, directly adsorbing the protein or the antibody to a real-time fluorescence quantification PCR (Polymerase Chain Reaction) tube wall to perform PLA detection; and mode 2, performing a tube wall solid phase PLA by a cross-linking agent fixing method, namely, cross-linking the protein or the antibody to the real-time fluorescence quantification PCR tube wall by glutaraldehyde to perform PLA detection. Specifically, the method comprises the following steps of: using a monoclonal antibody pab240, and sequentially establishing PLA detection of P53 protein (P53 protein mutant) and cTnI (cardiac troponin I). According to the method disclosed by the invention, the antibody is fixed on a PCR tube so as to perform protein PLA detection, magnetic beads are not needed in the detection process, defects caused by application of the magnetic beads are overcome, and the detection sensitivity to the protein is also improved greatly.

An assay method for determining the effect of Product R on virus infection of Hela cells. The method comprising the following step: (1) dividing Hela cells into several groups, (2) treating one group with Product R prior to infecting the cells with a virus and treating another group with Product R after the cells being infected with the virus, and (3) determining the effects of Product R on virus infection by comparing the changes in the cell cycle, DNA fragmentation and p53 protein in cells undergone the different treatments in step (2).

A method for determining if a subject with cancer or precancerous lesions or a benign tumor, will respond to treatment with an inhibitor selected from the group comprising an inhibitor of one or more enzymes in the mevalonate pathway, an inhibitor of geranylgeranyl transferase, an inhibitor of farnesyl transferase or an inhibitor of squalene synthase, by (i) obtaining a sample of the cancer cells, precancerous cells or benign tumor cells from the subject, (ii) assaying the cells in the sample for the presence of a mutated p53 gene or a mutant form of p53 protein or a biologically active fragment thereof, and (iii) if the cells have the mutated p53 gene or mutant form of the p53 protein, then determining that the subject will respond to treatment with the inhibitor or combinations thereof. Some embodiments are directed to treatment with the inhibitors.

The invention belongs to the field of biotechnology, and in particular relates to a method for regulating expression, quantity and activity of a heat shock protein 70 and application thereof. The method provided by the invention can also regulate tumor tissue, activity of a transcription factor NF-kB, expression and activity of a death receptor DR4 and DR5, phosphorylation and activity of JNK and c-Jun, expression and activity of a p53 protein, expression and proapoptotic activity of proapoptotic molecules Bax, Bid, t-Bid, capsase 3, capsase 8 and capsase 9, and expression and activity of c-FLIP and Bcl-2, and promote apoptosis of tumor tissues and cells. The invention also relates to application of the above method to preparation of drugs cooperatively applied with cell apoptosis induced drug.

More than 90% of mutations found in the p53 protein produce a conformational change in p53 which results in the exposure of an epitope, which is otherwise hidden in the hydrophobic core of the molecule. A single chain antibody (scFv) which specifically recognizes this common mutant epitope in mutant p53 but not in wild type p53 is disclosed. Also described are a DNA molecule encoding the scFv, pharmaceutical compositions comprising the antibody and methods of treatment using the pharmaceutical compositions.

The present invention is intended to provide a method for efficiently producing and providing a compound having a spirooxindole skeleton, for example, a compound having a spirooxindole skeleton and having antitumor activity that inhibits the interaction between Mdm2 protein and p53 protein, or an intermediate thereof, using an asymmetric catalyst. A compound having an optically active tricyclic dispiroindole skeleton is efficiently obtained through a catalytic asymmetric 1,3-dipolar cycloaddition reaction using ketimine as a reaction substrate and using a chiral ligand and a Lewis acid.

The invention provides peptides that can reactivate p53 mutants efficiently and specifically, as well as methods that allow the identification, selection and isolation of such peptides, in a precise, cost and time effective manner. In particular, there are provided mutant p53 reactivating peptides that can restore the native wild type p53 folding, and hence the tumor suppressor activity, to the mutant p53 protein. Such peptides are useful for treating various conditions and diseases in which p53 is mutated.

In fibrotic lung fibroblasts, basal levels of p53 protein (and miR-34a) are markedly suppressed, leading to reduced p53-mediated inhibition of uPA and uPAR, or concurrent induction of PAI-1. These changes contribute to excessive FL-fibroblast proliferation and production of extracellular matrix (ECM), and, therefore, pulmonary fibrosis. These processes are reversed by treating the cells, and treating subjects suffering from idiopathic pulmonary fibrosis (IPF) with the small organic molecule nutlin-3a (NTL) or with a peptide, CSP-4 (SEQ ID NO:1), or variants or derivatives or multimers of this peptide, which increase p53 levels by inhibiting MDM2-mediated degradation of p53 protein. Use of these compounds serves as a new approach to the treatment of IPF, they restore p53 expression and p53-mediated changes in the uPA-fibrinolytic system in FL-fibroblasts and restrict production and deposition of ECM.

The invention discloses an inhibitor by virtue of interaction of P53-MDM2 and applications of the inhibitor. The inhibitor by virtue of interaction of P53-MDM2 is ganoderic acid X, ganoderic acid Me, ganoderic acid Y, ganoderic acid T or ganoderic acid F or a composition formed by combining one of ganoderic acid X, ganoderic acid Me, ganoderic acid Y, ganoderic acid T and ganoderic acid F with pharmacologically acceptable additive components. The inhibitor by virtue of interaction of P53-MDM2, disclosed by the invention, is applied to promotion of the apoptosis of tumor cells and particularly applied to promotion of the apoptosis of the 95-D lung cancer cells expressing P53 proteins or H1299 lung cancer cells not expressing the P53 proteins; the inhibitor by virtue of interaction of P53-MDM2, disclosed by the invention, has the effects of remarkably inhibiting the activity of MDM2 in human malignant lung cancer cells, thereby inhibiting the degradation of the P53 proteins and further promoting the apoptosis of the tumor cells containing P53.

The invention discloses a fluorescence detection agent for a p53 protein. The fluorescence detection agent comprises a magnetic nano-particle, double-stranded DNA, and a functional layer coating the surface of the magnetic nano-particle and connected with the double-stranded DNA; the double-stranded DNA is obtained through hybridizing DNA-1 single strand and DNA-2 single strand; the DNA-1 is a carboxyl oligonucleotide probe, and the sequence is COOH-(CH2)6-TTT TTT TCC GGG CAT GCC CGG CAT TGC-3'; and the DNA-2 is the partial complementary sequence of the DNA-1, and the sequence is 5'-GGG CAT GCC C-3'. The invention also discloses a preparation method of the fluorescence detection agent, and an application of the fluorescence detection agent in the detection of the wild p53 protein. The agent has the advantages of low preparation cost, realization of sensitive, rapid and accurate capture of the p53 protein, realization of ultra-sensitive, highly- selective and highly-accurate detection of the p53 protein, low detection limit, and realization of quantitative analysis and detection of the p53 protein in normal cells and cancer cell lysate, and can be used in clinical diagnosis and early detection of cancers.

This invention belongs to the DNA recombination field in bioengineering. Wherein, with the recombination technique, inserting the gene fragment of B cell epitope composed by 37-46 peptide fragments on N-end of P53 protein into the carrier integrated with main coat protein gene of filamentous phage to form the new recombinant plasmid; in host, secreting the epitope gene product out of the cell to assemble in coat protein and form heterozygous protein that can be used as a antigen to detect antibody in clinical tumor patient serum (such as, gastric cancer, liver cancer, breast cancer and lung cancer).

The invention discloses a small interfering RNA (siRNA) for inhibiting canine p53 gene expression. The sequence of the siRNA is shown as SEQ ID No: 1 or SEQ ID No: 2 in a sequence table. The invention also discloses a DNA for coding the siRNA and a recombinant vector containing the siRNA or the DNA. Preferably, the recombinant vector is a slow virus vector for infecting a canine cell to obtain a canine cell model of p53 gene silence. The canine p53 gene siRNA sequence can effectively degrade mRNA of p53 genes so as to inhibit the expression of the p53 genes. By using the siRNA sequence for interfering with the expression of the canine p53 genes, on the one hand, a treatment medicament for diseases related with the p53 genes such as cell apoptosis related diseases can be prepared; and on the other hand, the established p53 knock down cell model has great significance for researching the effect of p53 protein on innate immunity of cells and possible specific mechanisms.

The invention belongs to the related fields of biomedical products and clinical detection and the like, and mainly relates to a new method for separating and purifying wild-type p53 protein. The separation and purification method of the wild-type p53 protein comprises cloning of a p53 gene, construction of recombinant expression plasmid, protein induced expression and purification, prokaryotic system-based inducible expression, ultrasonic cracking of thalli, and separation and purification of the p53 protein in the obtained cracked supernate by using nickel column affinity chromatography. High-concentration active target protein is separated and purified by the prokaryotic system-based inducible expression and the nickel column affinity chromatography in the ultrasonic cracked thalli supernate. Denaturation and renaturation of inclusion bodies are not performed, and the protein is directly purified from the ultrasonic cracked supernate, so that the activity of the target protein is furthest kept; and compared with the conventional method, the method has the advantage that: the yield of the target protein is greatly improved by optimizing the inducible expression and using a purification scheme.

The invention belongs to the technical field of tumor cells, and particularly relates to application of knocking out p53 genes in inhibiting proliferation and metastasis of glioma cells. According to technical scheme, the p53 genes in malignant glioma cells in a human body change by knocking out the p53 genes, thus p53 protein expression is reduced even completely deleted, and the purpose of inhibiting proliferation and metastasis of the glioma cells is achieved.

An isolated peptide is provided. The peptide comprises an amino acid sequence arranged in a space and configuration that allow interaction of the peptide with the DNA Binding Domain (DBD) of p53 through at least one residue of the DBD by which pCAP 250 (SEQ ID NO: 1) binds the DBD, wherein the peptide at least partially reactivates a mutant p53 protein, with the proviso that the peptide is not SEQ ID NO: 59-382.

The invention discloses a protein nano-drug for cancer targeted therapy and a preparation method thereof. The protein nano-drug is an exosome of humanized p53 protein transferred with covalent cross-linked triphenylphosphine (TPP). A protein nano-drug delivery system TPP / p53@Exos developed by the invention can recognize specific cancer cells in a targeted manner, efficiently inhibits the growth ofthe cancer cells by activating endogenous apoptosis pathways of the cells, has no toxic or side effect, and provides a new means for cancer treatment. The preparation method of the nano-drug deliverysystem has a high protein loading rate (about 75%), and can also be used for protection and targeted delivery of other protein drugs.

The invention discloses a recombinant protein G3P20-31, and a preparation method and an application thereof, belonging to the technical field of DNA recombination in bioengineering. According to the invention, a phage display technology is utilized to clone a gene segment for encoding a P53 protein N-terminal 20-31 peptide fragment into a filamentous phage vector PIII protein gene, and a recombinant protein for displaying the P53 protein epitope is prepared and displayed onto the surface of bacteriophage, so the recombinant protein is used for detecting the serum p53 antibody of a tumor patient. The invention also discloses an application of the recombinant protein G3P20-31 in preparation of a biological product for detecting a tumor patient serum p53 antibody. The biological product has the advantages of strong specificity, high sensitivity, simple preparation, low cost and the like.