54 results about "Connexon" patented technology

The invention relates to a PEG interleukin 15 conjugate, namely PEG interleukin 15. PEG and interleukin 15 are connected on an N terminal amino group through or not through small peptide connexon to obtain the uniform PEG interleukin 15 conjugate. The activity of the original interleukin 15 can be at least reserved in vivo, and the PEG interleukin 15 conjugate has longer half-life period and average time to peak.

Disclosed are a type of tridentate linkers and use thereof. The tridentate linkers can be used to prepare an antibody drug conjugates as represented by formula I,L-(T-A-D)n   Iwherein, L is an antibody, antibody fragment or protein; T is a tridentate linker part; A is a cleavable linker group or a noncleavable linker part; D is a drug part; n is an integer of 0-8. The structure of the tridentate linker part is as represented by formula II.wherein, W is substituted aryl, heteroaryl, linear alkyl, cycloalkyl, heterocycloalkyl, or any combination thereof.

The invention discloses a surface self-assembly gold nanoprobe with the free radical capture performance and a preparing method and the application thereof. The surface self-assembly gold nanoprobe with the high free radical capture performance is prepared by the following steps: the surface of a gold nano-particle is connected with nitrone free radical probe ligand with the structure of formula II by a connexon of the structure of formula I. The nitrone free radical probe ligand is connected with R3 radical group end. The gold nano-particle is connected with sulfhydryl end of the connexon. In formula Ia and formula Ib, R3 is -OH, -NH2 or -COOH, n is equal to 2-11, and n1 is equal to 1-4. In formula IIc, R1 is -P(O)(OC2H5)2, and R2 is OH, COOH or CH2OH. The surface self-assembly gold nanoprobe with the high free radical capture performance can capture free radicals under low concentration and can be applied as an antioxidant.

The invention discloses a cysteine-modified antibody-toxin conjugate. The conjugate is prepared by the steps: modifying heavy chain 235th serine (S) of a target antibody into cysteine (C), and performing fixed point coupling on free sulfydryl (-SH) of the modified cysteine and a mc-vc-PAB-OH connexon coupled with small-molecule high-activity cytotoxin (Payload), thereby obtaining the cysteine-modified antibody-toxin conjugate with excellent uniformity. A toxic-antibody ratio (DAR) is 1.6-2.0. The antibody-toxin conjugate has a general formula of 2C2-HC-S235-mc-vc-PAB-payload. Meanwhile, the invention further discloses application of a method for preparing and purifying the TDC medicine in treatment of EGFRvIII over-expressed tumors.

The invention discloses a novel cathepsin B activation type targeted anti-tumor polypeptide m(KLA)-iRGD, the molecular weight is 2570, and the amino acid sequence is D(KLAKLAKKLAKLA)KGGCRGDKGPDC. The polypeptide is formed by KLA peptide and iRGD peptide through flexible connexon Gly-Gly. The sequence of the KLA peptide is KLAKLAKKLAKLAK, and amino acids of the KLAKLAKKLAKLA sequence at the N terminal of the polypeptide are all D-amino acids. The sequence of the iRGD peptide is CRGDKGPDC. The polypeptide has the advantage that the tumor targeted peptide iRGD and the apoptosis promoting peptide KLA are connected into the new polypeptide m(KLA)-iRGD for the first time. The polypeptide m(KLA)-iRGD can be specifically combined with alphaVbeta3 and NRP-1 receptors and enter tumor cells, mitochondrial apoptosis is initiated after the polypeptide is activated by intracellular cathepsin B (CTSB), tumor cells are selectively killed, and growth and migration of tumors are remarkably inhibited. Under the same concentration, a polypeptide m(KLA)-iRGD comparison peptide has higher tumor targeting and higher capacity of inhibiting tumor growth and migration. Meanwhile, the polypeptide m(KLA)-iRGD has no obvious toxicity on mouse weight and vital organs, and a solid foundation is laid for further clinical research and application.

The invention adopts oligonucleotide aptamers for identifying special tripeptide sequence KAI, GEL, DGI and LAS, extanding oligonucleotide aptamers by Structure-switch and phosphorylation decorating 5' end thereof, synthetising four connexons; connecting oligonucleotide aptamers combinated with target albumen through proximity dependent DNA Ligation technique as chain ringlike DNA; amplifying detecting signals via rolling-circle DNA duplicate method; the technique can detect target albumen sensitively, the least detecting concentration can get nM level; this method with flexibility can adjust according needed numbers of ligand; can detect albumen composite, provide sensitive detecting method for chip technique using oligonucleotide aptamers as ligand.

The invention discloses a small-molecule modified target paclitaxel precursor medicament, as well as a preparation method and application thereof. The molecular structure of the precursor medicament comprises folic acid, amino acid and paclitaxel, wherein the folic acid is used as a targeting ligand, and the amino acid is used as a connexon. The precursor medicament also comprises a fluorescent dye. Compared with the prior art, the precursor medicament has the advantages that the problem of water solubility and targeting of paclitaxel can be solved, the toxicity of the paclitaxel medicament can be reduced, the treatment effect of the paclitaxel medicament can be improved, body immunogen toxin caused by macro-molecular protein can be avoided, and the defects of poor encapsulation rate, unstable structure and the like caused by using lipidosome as an insoluble medicinal carrier can be overcome.

The invention provides a double-substituted maleic amide connexon coupled with an antibody as well as a preparation method and application of the connexon. Particularly, a strong cytotoxic activity substance and biological macromolecules are coupled through a kind of new connexon. The kind of connexon can selectively act with a disulfide chain at the same time, so that the substance homogeneity and stability of a conjugate are greatly improved. The conjugate prepared from the connexon has high inhibitory activity for cell strains which express corresponding antigens. The invention further provides a preparation method and application of the conjugate.

The invention discloses a preparation method of a nucleic acid targeted capture sequencing library based on a long chain molecule inversion probe. The preparation method comprises the following steps:a, synthesizing a capture probe A, a capture probe B and a connexon C; b, adding phosphorylated probes A and B and the connexon C into a ligase reaction system, simultaneously adding DNA (Deoxyribonucleic Acid) ligase so as to connect the A with the B under the bridging effect of the C; c, combining multiple connection mixtures for different target areas, and separating and purifying the connected product by denaturing electrophoresis or a nucleic acid purification kit to obtain the long chain molecular inversion probe; d, mixing the long chain molecular inversion probe with DNA or cDNA of ato-be-tested sample, hybridizing, adding DNA polymerase, DNA ligase, dNTP and a Mg2<+>-containing buffer solution into a buffer solution, extending the long chain molecular inversion probe and formingclosed molecules under the action of the DNA ligase; e, adding exonuclease to degrade non-cyclic DNA molecules; f) carrying out PCR (Polymerase Chain Reaction) amplification by using primers corresponding to a common sequence region of the long-chain molecular inversion probe to obtain the sequencing library of a targeted region.

The invention discloses a coding method of a nucleic acid sequence to be detected which is a method of simultaneously analyzing high throughputs of nucleic acid sequences of different samples or different nucleic sequences of one sample and in particular relates to a coding and decoding method of the nucleic acid sequence to be detected. The scheme of the invention is to respectively cut every nucleic sequence to be detected into nucleic acid fragments and then respectively connect one connexon with the two ends of the nucleic acid fragment to form templates. One connexon consists of an amplified (complementary to a sequencing primer) sequence fragment and a section of sequence codes. And the sequence codes on the nucleic fragment of the same nucleic sequence are same. Thus, the nucleic acid sequences of the codes are formed. The sequence codes consists of four bases of A, T, C or G and are obtained through a synthetic method. The invention can solve the contradict between the present high-energy DNA sequence detection technology and the low throughput of samples to be detected by users with multiple samples and lower the detection cost. The method is simple.

The invention adopts oligonucleotide aptamers for identifying special tripeptide sequence KAI, GEL, DGI and LAS, extanding oligonucleotide aptamers by Structure-switch and phosphorylation decorating 5' end thereof, synthetising four connexons; connecting oligonucleotide aptamers combinated with target albumen through proximity dependent DNA Ligation technique as chain ringlike DNA; amplifying detecting signals via rolling-circle DNA duplicate method; the technique can detect target albumen sensitively, the least detecting concentration can get nM level; this method with flexibility can adjust according needed numbers of ligand; can detect albumen composite, provide sensitive detecting method for chip technique using oligonucleotide aptamers as ligand.

The invention provides an anterograde monosynaptic transneuronal virus tracing system, which is drawing a direct post-synaptic target of neuron of special type of a specific brain nucleus. The anterograde monosynaptic transneuronal virus tracing system comprises recombinant defective HSV-1 viruses derived from H129 and recombinant AAV9 helper viruses (AAV9-TK-GFP and AV9-DIO-TK-GFP), wherein the recombinant defective HSV-1 viruses comprise an integrated first expression cassette; the first expression cassette comprises a first promoter, a first fluorescent protein encoding sequence and a resistance peptide encoding sequence, wherein the first expression cassette completely or partially replaces the gene sequence of thymidine kinase (TK), and a TK function (H129-[delta]TK-tdT) becomes lost in the generated H129 derived recombinant defective HSV-1 H129 viruses; and the recombinant AAV9 helper viruses comprise an integrated second expression cassette; the second expression cassette comprises a second promoter, a TK encoding sequence, a connexon peptide encoding sequence and a second fluorescent protein encoding sequence, wherein through TK expression of the second expression cassette, the replication of the H129 derived recombinant defective HSV-1 viruses is achieved, wherein different fluorescent proteins are encoded by virtue of the first and the second fluorescent protein encoding sequences.

The invention discloses an HCV recombinant fusion antigen and an expressed gene and a preparation method thereof. Amino acids of the HCV recombinant fusion antigen are successively composed of an amino acid shown in SEQ ID No. 1, a first connexon, an amino acid shown in SEQ ID No. 2, a second connexon, an amino acid shown in SEQ ID No. 3, a third connexon and an amino acid shown in SEQ ID No. 4, wherein each connexon is composed of six flexible, polar and hydrophilic amino acids with small molecular weight. The preparation method comprises the following steps: constructing a recombinant plasmid pET41a-Core-NS3-NS41-NS42; and expressing and purifying the HCV recombinant fusion antigen. The HCV recombinant fusion antigen has high sensitivity and specificity and is applicable to the field of biological medicine, especially to colloidal gold test strips for antibodies to hepatitis c.

The invention discloses a drug conjugate with multiple targeting ligands having a cell endocytosis mediating function. The drug conjugate consists of drug molecules and connexons coupled with the drug molecules; the drug conjugate at least comprises two or more ligands which can be specifically bound to cell surface receptors; the ligands at least include one first ligand which has the cell endocytosis mediating function; and at least one of the ligands and the first ligand are coupled and then are selectively coupled with the drug molecules or the connexons so as to form the drug couplet. The drug conjugate provided by the invention has the beneficial effects that by combining targeting and endocytosis structures, polypeptide, which is targeted by any ligands, is taken as a guiding part, so that the targeting scope of the like medicines is widened. The affinity and the targeting performance of the drug conjugate on diseased cells can be enhanced by virtue of the multiple targeting ligands, so that efficient toxin drugs such as MMAE can be carried, efficacy can be enhanced, a therapeutic window can be widened and side effects of the medicines can be avoided.

The high efficiency full genome fixing method relates to method of amplifying full genome in high efficiency and preparing full genome DNA chip. The method includes: 1. machining pores in one flat hard material and fixing same oligonucleotide molecules; 2. cutting genome nucleic acid with enzyme into 50-1000 base size fragments and connecting the genome fragment nucleotide sequences with one pair of common connexons under the action of enzyme to form genome fragment nucleotide sequences containing connexons; 3. heat denaturing and diluting the genome fragment nucleotide sequences containing connexons and adding to the pores; and 4. adding amplifying reaction liquid into the pores and amplifying the genome fragment nucleotide sequences containing connexons to realize exponential amplification of the full genome sequence in fragment form.

The invention discloses a bioconjugation method for an IgG4 antibody. The bioconjugation method for the IgG4 antibody comprises the following steps of: step 1, reduction: the IgG4 is reduced, so fourpairs of interchain disulfide bonds in the IgG4 antibody are all made open; step 2, partial oxidation: the four pairs of open interchain disulfide bonds in the IgG4 antibody are partially oxidized, and the number of the free mercapto groups of the obtained IgG4 antibody reach the targeted average conjugation number of 4+ / -1; step 3, a coupling reaction: bioconjugation is conducted by adopting thefree mercapto groups of the IgG4 antibody and a connexon-conjugate complex; and step 4, purification of bioconjugation products: excess conjugates in the bioconjugation products which are obtained inthe step 3 are removed so as to obtain the final product. The bioconjugation product mixture obtained by using the method has better product distribution, the content of the conjugation products witha linker number of 0 is less than 5%, and the content of the conjugation products with a linker number of 8 is less than 15%; after routine purification is conducted on the obtained bioconjugation product mixture, the final product can be applied to animals or clinical trials directly.

The invention relates to an industrialized production method for preparing an antibody coupling medicine Adcetris connexon. Through a method for protecting a hydroxyl group of a compound IV, the moisture absorption performance is improved, the post-processing operation is simplified, and the product meeting requirements of pharmaceutical-grade raw material drugs is obtained. The method is not only suitable for a small amount of preparation in laboratories but also suitable for large-scale industrial production.

It is intended to efficiently inject into a target cell, a substance charged within a liposome. The present inventors have found that connexin synthesized within a liposome is introduced as connexon having a gap junction function into the liposome membrane. Specifically, the liposome according to the present invention is a liposome in which connexon composed of connexin synthesized by an in-vitro protein synthesis system is incorporated in a state of having a gap junction function.

The invention relates to a recombinant plasmid for coding a novel fusion protein for a human HMGB1 A box and an acidic tail. An HMGB1 B box is deleted between the HMGB1 A box and the acidic tail; and the HMGB1A box is connected with the acidic tail through a flexible connexon. The invention also provides a preparation method of the recombinant plasmid and the novel fusion protein secreted and expressed by the recombinant plasmid. The fusion protein deletes an inflammatory functional domain B box, fully uses two beneficial functions of the antibacterial activity of HMGB1 and the obvious anti-inflammatory effects of the A box, and effectively avoids the side effect of inflammatory effect at the same time. The fusion protein can effectively resist the release of various inflammatory cytokine induced by the HMGB1, has stronger anti-inflammatory activity than the single A box does, and can inhibit growth of bacteria at the same time.

The invention discloses an antibody drug conjugate with alkenylsulphonylamino as a connexon and application of the antibody drug conjugate. A preparation method of the antibody drug conjugate comprises the steps of connecting amino and alkenylsulphonyl on the structure of a drug molecule to obtain an alkenylsulphonylamino derivative of the drug molecule; and then, coupling the drug molecule and an antibody compound through carrying out Michael addition reaction on alkenyl on the derivative of the drug molecule and amino on the antibody compound. The alkenylsulphonylamino derivative of the drug molecule can be selectively subjected to coupled reaction with amino on an antibody, conjugates can be selectively and efficiently subjected to coupled reaction with amino acid residues on the surface of the antibody in a water phase, and meanwhile, the activity of the antibody can also be kept. The obtained antibody drug conjugate has high stability in a chemical environment and an in-vivo environment. In addition, the antitumor activity of the antibody drug conjugate is improved as comparison with a single drug molecule.

The invention relates to a P-5m-Fc fusion protein and an expression gene thereof, and a preparation method and applications of the P-5m-Fc fusion protein, and belongs to the technical field of medicine and biology. The P-5m-Fc fusion protein is formed by linking a functional unit and the Fc portion of human immunoglobulin IgG1, wherein the functional unit is formed by linking an amino acid sequence represented by SEQ ID NO.1 and a connexon, the connexon is a polypeptide formed by six sequentially-linked glycines, the P-5m-Fc fusion protein has at least one functional unit, and when the P-5m-Fc fusion protein has a plurality of the functional units, the plurality of the functional units are sequentially linked, and then are linked to the Fc portion of immunoglobulin. According t the present invention, with the P-5m-Fc fusion protein, P-5m and the Fc portion of the human IgG1 are subjected to fusion expression to prepare the peptide, such that the efficacy of the P-5m octapeptide can be increased, the half-life of the drug can be significantly prolonged, the good patent medicine property is obtained, and the experiment results prove that the significant anti-tumor-metastasis function is provided.

The invention discloses a method for detecting and identifying Lactobacillus acidophilus NCFM. Fragments of to-be-detected strain 16SrDNA are subjected to PCR amplification by means of primers plb16A,mlb16A, plb16B, mlb16B, plb16X, mlb16X and high-fidelity DNA polymerase; an amplification product is digested by restriction endonucleases AhdI, BmrI and XcmI, purified and connected with a T vector;a connecting product is transformed into Escherichia coli, connexons are screened, plasmids are extracted for sequencing and comparison, and whether the detected strain is the Lactobacillus acidophilus NCFM is identified preliminarily. The method has the following advantages: the Lactobacillus acidophilus can be identified by means of the length of the amplification product; the Lactobacillus acidophilus can be identified by means of the length of the amplification product after being digested by AhdI, BmrI and XcmI; the amplification product is obtained through amplification by high-fidelityDNA polymerase, so that the mispairing rate of the amplification product is low, sequencing and comparison are performed after the amplification product is connected with the T vector, and the Lactobacillus acidophilus NCFM can be identified preliminarily by means of a result.

The invention discloses a variant of tumor necrosis factor-related apoptosis-inducing ligand. The variant is a fusion protein of the tumor necrosis factor-related apoptosis-inducing ligand and F3 peptide, wherein the F3 peptide is connected with the N terminal or C terminal of the tumor necrosis factor-related apoptosis-inducing ligand through connexon. The invention also discloses a nucleotide sequence, a recombinant vector and recombinant bacterium containing the nucleotide sequence, and a preparation method and application of the variant. According to the invention, the variant of the tumor necrosis factor-related apoptosis-inducing ligand is prepared by using a genetic engineering manner; the variant is obviously superior in affinity to tumor cells, capability of inducing apoptosis of tumor cells, tumor targeting performance and in-vivo antineoplastic activity compared with the tumor necrosis factor-related apoptosis-inducing ligand; and the variant has excellent curative effect on tumors and has good clinical application prospects.

The invention belongs to the technical field of biological medicines, and particularly relates to an alpha-synuclein targeting compound as well as a preparation method and application thereof. The compound simultaneously contains an alpha-synuclein ligand, a connexon and an E3 ligase ligand structural fragment, and can recruit E3 ligase in cells to carry out ubiquitination modification on alpha-synuclein, so that chemical induced degradation of the alpha-synuclein is realized by virtue of UPS (Uninterrupted Power Supply); and tests prove that the compounds can achieve the effects of remarkably reducing the intracellular alpha-synuclein aggregate level and improving the cell viability by targeting alpha-synuclein and inducing ubiquitination of the alpha-synuclein, and have better application prospects in the field of treatment of alpha-synuclein related diseases.

The invention discloses a polypeptide molecular imaging probe targeting mesenchymal stem cells, a preparation method thereof and mesenchymal stem cells marked by the probe. The molecular probe includes: a polypeptide for identifying mesenchymal stem cells and Contrast unit metal Gd complexes for enhanced contrast in magnetic resonance imaging. A main feature of this molecular probe is that a polypeptide probe binds 1 to 4 metal Gd complex molecules through dendrimers, and spacers and linkers are used to improve the complexation of polypeptides with dendrimers and dendrimers with Gd space between objects. The polypeptide molecular imaging probe can significantly enhance the contrast of magnetic resonance weighted imaging.

The invention relates to an intermediate (a combination of a connexon and a load) for preparing an antibody and a drug conjugate (ADC) and a preparation method and application thereof. Specifically, the combination of the connexon and the load has a structure shown as a formula I, the compound shown as the formula I can be applied to coupling of high-hydrophobicity compound loads such as paclitaxel (PTX') and the like which are not easy to couple, and the prepared ADC molecule has good hydrophilicity and thermal stability and can reach a relatively high drug-to-antibody ratio (DAR) value. According to the intermediate, the Trop-2 targeting ADC prepared by using the connexon represented by the formula I shows the strong in vitro drug effect in the low expression cell line in the Trop-2 compared to the Trop-2 targeting ADC with the effective loads of MMAE and SN38, and shows the strong tumor inhibition ability in the mouse in vivo xenotransplantation tumor model, and the bystander effect is proved. Experimental results show that the problem that the paclitaxel compound is used as an ADC effective load can be solved by utilizing the hydrophilic connexon, and ADC molecules which are expected to be used for treating various human cancers are obtained.

The full genome rolling circle amplification and its product fixing method realizes the simultaneous acquisition of full genome information to save template preparing time greatly, and the simultaneous amplification of the full genome information after fragmentation in single molecule form can raise the detection sensitivity while maintaining the number of single molecules after fragmentation. The genome DNA is enzyme digested or ultrasonically broken into fragments, the genome DNA fragments are connected with connexons to form circle and amplified with the circular molecule to obtain rolling circle amplified genome fragmented nucleic aid sequence. The nucleic aid sequence is diluted and fixed onto the surface of hard carrier chemically to realize the fixation of full genome sequence and constitute full genome DNA chip.