55 results about "3c protease" patented technology

The invention relates to valerolactam enterovirus 71 (EV71) 3C protease inhibitors, wherein a structure general formula of the inhibitors is represented by a compound M, various variables in the structure is defined in an instruction, and EV71 replication is effectively inhibited or blocked with the compounds. The present invention relates to discoveries and applications of the compounds containing the structure represented by the formula (M), optical isomers, metabolites with pharmaceutical activities, pharmaceutically acceptable salts, solvates, and prodrugs thereof in preparations of anti-virus drugs for treatment of hand-foot-mouth disease infection. The present invention further relates to an intermediate of the structure compound represented by the formula (M) and a synthesis method thereof.

The invention relates to a caprolactam anti-enterovirus 71 (EV71) 3C protease inhibitor with a structural formula shown as compounds (M). Each variable in the structure is defined as the specification. The compounds can effectively inhibit or block replication of enterovirus 71. The invention relates to discovery and application of compounds comprising a structure of formula (M), various optical isomers thereof, metabolites with pharmaceutically activity, pharmaceutically acceptable salts, solvates and prodrugs in preparing antiviral drugs for treating virus infections of hand-foot-and-mouth diseases. The invention also relates to an intermediate and a synthetic method for preparing the compounds having the structure of the formula (M).

This application is directed generally to foot-and-mouth disease virus (FMDV) 3C proteases that have been modified by mutating a polynucleotide sequence coding for the FMDV 3C protease. The modified FMDV proteases exhibit proteolytic activity on FMDV P1 precursor protein and exhibit a reduction in one or more toxic or inhibitory properties associated with an unmodified FMDV 3C protease on a host cell used to recombinantly produce it. Vectors carrying polynucleotides encoding modified FMDV 3C protease sequences can induce production of FMDV virus-like particles in a host cell when expressed in the host cell. The modified FMDV 3C proteases can generally be used to produce immunogenic FMDV preparations capable of inducing an immune response against FMDV.

A structure general formula of a caprolactam aldehyde enterovirus 71 (EV 71) 3C protease inhibitor is as shown in chemical combination A, all variables in the structure are defined in an instruction book, and the compound effectively restrains or blocks the copy of the enterovirus 71. The invention relates to a compound with the structure as shown in formula (A), various types of optical isomers of the compound with the structure as shown in formula (A), drug active metabolites, officinal salt, solvate and discovery and application when the compound with the structure as shown in formula (A), the various kinds of optical isomers of the compound with the structure as shown in formula (A), the drug active metabolites and the officinal salt are used for preparing antiviral drugs for curing hand-foot-mouth virus infection diseases. The invention further relates to an intermediate and a synthetic method of preparing the compound with the structure as shown in formula (A).

The present invention relates to a class of novel Michael receptor-based virus 71 (EV71) 3C protease inhibitors, wherein various variables of the structure general formula (M) are defined in the specification, and the compounds effectively inhibit or block the replication of enterovirus 71. The present invention relates to discovery and applications of the compound containing the structure generalformula (M), various optical isomers, pharmaceutically active metabolites, pharmaceutically acceptable salts, solvates and prodrugs thereof in preparation of antiviral drugs for the treatment of hand-foot-mouth virus infection diseases. The invention relates to an intermediate and a synthesis method for preparing the compound having the structure general formula (M). The formula (M) is defined inthe specification.

The invention relates to an efficient fusion protease, namely an ELP-3C protease which is obtained through the fusion expression between elastin-like polypeptide (ELP) and human rhinovirus 3C protease. As the ELP is adopted as a purification tag, the fusion protease can be purified without the assistance of a chromatography separation technology, thereby reducing the cost of purification. The ELP-3C protease can be applied to cutting fusion protein; and after enzyme cutting of substrate protein, the ELP-3C protease is triggered to precipitate through simply increasing the concentration of salt ions and is removed through centrifuging, wherein the process is performed without the assistance of the chromatographic separation technology, and the ELP-3C protease can be used for the enzyme cutting continuously for multiple times and keeps the enzyme activity after multiple times of the enzyme cutting, thus the cost of large-scale protein purification can be reduced.

Novel 1,2,4-thiadiazole compounds are provided, which are effective as inhibitors of cysteine activity-dependent enzymes and in particular of cysteine proteases. The compounds are useful in treating acne by inhibition of transglutaminase, common cold by inhibition of human rhinovirus 3C protease and inflammatory joint disease by inhibition of cathepsins. The compounds of the present invention are 3,5-disubstituted 1,2,4-thiadazole of the general formula (I):where Z is a nitrogen containing group with recognition sequence for the enzyme and Y is a substituent that tunes the reactivity of the inhibitor towards the thiol group of the cysteine activity-dependent enzyme. The Y group may also serve in recognition.

The invention discloses a structure and application of an Enterovirus 71 (EV71) 3C protease and belongs to the field of RNA virus protein. The invention also discloses an application of the 3C protease and a substrate binding groove thereof in drug design. The protein provided by the invention has a special substrate binding groove and new EV71 virus 3C protease drugs are designed according to the spatial structure of the substrate binding groove, thus the inhibitors aiming at the EV71 virus 3C protease, which are more specific and have better effect, can be obtained, potential drug candidates can be provided for the clinical treatment of hand, foot and mouth disease and the EV71 virus 3C protease has very high application value.

Polynucleotide constructs that express an engineered foot-and-mouth disease (FMDV) P1 precursor protein and a non-FMDV TEV protease and methods for safe and efficient recombinant production of FMDV antigens and immunogens. Recombinant production of FMDV antigens avoids the need to culture highly-infectious FMDV, while conventional culture methods for producing FMDV antigens rely on the native FMDV 3C protease which exerts toxic effects on host cells. The inventors have developed a new system that efficiently and safely processes FMDV P1 precursor without the FMDV 3C protease, thus avoiding the toxic effects associated with use of the 3C protease. The invention is also directed to the FMDV antigens and virus-like particles produced by this system as well as to FMDV vaccines, diagnostics and other biologics.

The invention discloses a 4-iminooxazolidine-2-ketone enterovirus 71(EV71) 3C protease inhibitor of which the structural general formula is shown as a compound (M), each variable in the structure is defined as the specification, and the compounds are used for effectively inhibiting or stopping the replication of enterovirus 71. The invention relates to discovery and application of a compound with a structure shown as the formula (M) as well as various optical isomers of the compound, a pharmaceutically active metabolite, a medicinal salt, a solvate and a prodrug of the compound in preparation of antiviral drugs for treating hand-foot-mouth virus infection diseases and also relates to an intermediate for preparing the compound with the structure shown as the formula (M) and a synthesis method.

The invention discloses an application of asymmetric aryl disulfide compounds serving as virus 3C protease inhibitors in preparation of antiviral drugs. The structural formula of the compounds is shown in the formula I or the formula II, wherein in the formula I, R1 represents mono-substituted groups or poly-substituted groups on a benzene ring and is independently selected from -NO2, C1-C3 alkyl, C1-C3 alkoxy, R'OCO- and a condensed ring constructed through direct condensation with the benzene ring, R' represents C1-C3 alkyl, R2 represents H or acyl with the number of carbon atoms being 1-3, and R3 represents H or C1-C3 alkyl; in the formula II, X represents NH or S, R4 represents mono-substituted groups or poly-substituted groups on the benzene ring, R1 is independently selected from -NO2, C1-C3 alkyl and halogen, R5 represents H or R''OCO-, R'' represents C1-C3 alkyl, R6 represents H or R''' CONH-, and R''' represents C1-C3 alkyl. The asymmetric aryl disulfide compounds can have an inhibition effect on CVB3 viruses on the cell level by inhibiting the activity of CVB3 3C and has good antiviral drug application prospect.

Novel 1,2,4-thiadiazole compounds are provided, which are effective as inhibitors of cysteine activity-dependent enzymes and in particular of cysteine proteases. The compounds are useful in treating acne by inhibition of transglutaminase, common cold by inhibition of human rhinovirus 3C protease and inflammatory joint disease by inhibition of cathepsins. The compounds of the present invention are 3,5-disubstituted 1,2,4-thiadazole of the general formula (I): where Z is a nitrogen containing group with recognition sequence for the enzyme and Y is a substituent that tunes the reactivity of the inhibitor towards the thiol group of the cysteine activity-dependent enzyme. The Y group may also serve in recognition.

The invention discloses a recombinant CL7-CVN protein, and a preparation method and applications thereof. The preparation method includes cloning a coding sequence on an expression vector to obtain arecombinant expression vector; and converting the recombinant expression vector into host cells to perform expression and purification so as to obtain the recombinant CL7-CVN protein. Through a synthesized CVN gene and gene assembly method, the fusion expression vector can be built, and the highly soluble expression of the vector in escherichia coli can be realized; and according to the heat resistance of the recombinant CL7-CVN protein and through heat treatment and 3C protease digestion, the recombinant CL7-CVN protein and CVN protein can be rapidly obtained through one-step affinity chromatography, and purity can reach 99%. The provided preparation method can make the recombinant protein expression significantly enhanced, make activity more stable and make a purification method simpler;and both the prepared recombinant CL7-CVN protein and CVN protein have antiviral activity.

The invention discloses a foot-and-mouth disease virus (FMDV) 2C3ABC recombinant protein as well as a preparation method and application thereof, and belongs to the field of pharmaceutical biotechnology. According to the invention, the mutation of the following sites is performed on the basis of an original FMDV 3C protease gene: Cys142-Ser, Cys163-Gly. FMDV recombinant protein 2C3ABC is expressed as an inclusion body in the bacterial cytoplasm, and is subjected to separation, denaturation, renaturation and multi-step purification, to obtain a complete and enzymolysis-free FMDV nonstructural protein mu2C3ABC, wherein the protein has solubility and complete antigenicity, and has a molecular weight of 72 kDa. The protein, as a diagnostic antigen, is prepared into chromatographic strips, has sensitivities of 98.4% and 100% respectively in the detection of FMDV experimentally infected pigs and naturally infection-free pigs, and has specificities of 100% and 98% respectively in the detection of naturally infection-free pigs and vaccine-immunized pigs. Compared with commercial kits Ceditest and UBI, the FMDV 2C3ABC recombinant protein has a high degree of consistency, can be used to distinguish infected animals and immunized animals, wherein K = 0.823 (p is smaller than 0.05).

The invention relates to a caprolactam anti-enterovirus 71 (EV71) 3C protease inhibitor with a structural formula shown as compounds (M). Each variable in the structure is defined as the specification. The compounds can effectively inhibit or block replication of enterovirus 71. The invention relates to discovery and application of compounds comprising a structure of formula (M), various optical isomers thereof, metabolites with pharmaceutically activity, pharmaceutically acceptable salts, solvates and prodrugs in preparing antiviral drugs for treating virus infections of hand-foot-and-mouth diseases. The invention also relates to an intermediate and a synthetic method for preparing the compounds having the structure of the formula (M).

Compounds of formula (I), wherein the variables of the formula are as defined in this disclosure, can advantageously inhibit or block the biological activity of picornavirus 3C protease. These compounds, as well as pharmaceutical compositions containing these compounds, are suitable for treating patients or hosts infected with one or more picornaviruses, such as RVP. Intermediates and synthetic methods for the preparation of such compounds are also described.

The invention relates to a recombinant DNA vector for efficiently preparing foot and mouth disease virus-like particles, the foot and mouth disease virus-like particles prepared by utilizing a recombinant expression plasmid vector containing the recombinant DNA vector to transfect a host cell, application thereof and a vaccine. Aiming at the problems of toxicity of 3C protease to the host cell andimpact of 3C protease on yield of foot and mouth disease VLPs, a 3CshRNA segment having disturbing effect on gene expression of 3C protease is designed, so that expression of 3C protease in the process of preparing the foot and mouth disease VLPs is reduced obviously; three sites are mutagenized on the basis of original foot and mouth disease virus 3C protease gene, so that action effect of 3C protease is regulated specifically. Through modifying the 3C protease gene, the toxicity of 3C protease to the host cell is lowered, and effective cutting performance of 3C protease to foot and mouth disease virus P1 precursor protein is improved, so that synthesis yield of the foot and mouth disease VLPs is increased greatly, and a foundation is laid for large-scale production and use of foot and mouth disease VLP vaccines.

The invention relates to the technical field of medicines and in particular relates to a hand-foot-and-mouth disease resistant drug activity detection method and kit. The detection method comprises the following steps: inserting a nucleotide sequence encoding Gln-Gly into a luciferase plasmid, thereby obtaining recombinant luciferase containing Gln-Gly by virtue of in-vitro acellular transcription and translation; performing in-vitro recombinant expression on hand-foot-and-mouth virus protease, wherein the hand-foot-and-mouth virus protease refers to 2A or 3C protease; carrying out an enzymatic reaction between the hand-foot-and-mouth virus protease, the recombinant luciferase containing Gln-Gly and a test substance, detecting the bioluminescence intensity of fluorescein, thereby obtaining the hand-foot-and-mouth disease resistant drug activity of the test substance. The detection method disclosed by the invention has the advantages of simplicity, trace amount and rapidness, is small in drug screening workload, rapid and accurate in experiment and high in experimental result repeatability and can be used for screening and developing hand-foot-and-mouth disease resistant drugs. The method refers to a bioluminescence detection method based on luciferase, the sensitivity is high, a few interference factors exist, and the compatibility of the test substance is high.

The invention utilizes a yeast endoplasmic reticulum sequestration screening system (YESS) to quickly analyze the specificity of a cleavage site of a substrate of protease, and finally provides a cleavage efficiency enhanced substrate sequence mutant of HRV (Human Rhinovirus) 3C protease and application thereof. The substrate mutant includes an amino acid sequence as shown in SEQ ID NO.1 in a sequence table, and has a better practical application advantage in the removal of a purification tag in a protein purification process, and therefore, the further application and research of the HRV 3C protease are facilitated.

Peptido and peptidomimetic compounds of the formula: wherein the formula variables are as defined in the disclosure, advantageously inhibit or block the biological activity of the picornaviral 3C protease. These compounds, as well as pharmaceutical compositions containing these compounds, are useful for treating patients or hosts infected with one or more picornaviruses, such as RVP. Intermediates and synthetic methods for preparing such compounds are also provided.

The invention discloses a structure of a coxsackie virus A16-3C protease and an application thereof, which belong to the field of RNA (Ribose Nucleic Acid) virus proteins. The invention also discloses an application of using a 3C protease and a substrate binding groove in drug design. The protein provided by the invention has a special substrate binding groove; a novel anti-virus CVA16 3C protease drug can be designed according to the space structure of the substrate binding groove so as to obtain a more specific inhibitor with better effect for the virus CVA16 3C protease; therefore, a potential alternative drug is provided for clinically treating hand-foot-and-mouth diseases and very high application values are obtained.

Peptido and peptidomimetic compounds of the formula: wherein the formula variables are as defined in the disclosure, advantageously inhibit or block the biological activity of the picornaviral 3C protease. These compounds, as well as pharmaceutical compositions containing these compounds, are useful for treating patients or hosts infected with one or more picornaviruses, such as RVP. Intermediates and synthetic methods for preparing such compounds are also provided.

Peptido and peptidomimetic compounds of the formula: wherein the formula variables are as defined in the disclosure, advantageously inhibit or block the biological activity of the picornaviral 3C protease. These compounds, as well as pharmaceutical compositions containing these compounds, are useful for treating patients or hosts infected with one or more picornaviruses, such as RVP. Intermediates and synthetic methods for preparing such compounds are also provided.

The present invention relates to a recombinant adenoviral vector for generating immunity against enterovirus infection. In one embodiment, the recombinant adenoviral vector of the invention comprises an expression cassette encoding a PI protein and a 3 CD protease of an enterovirus. In another embodiment, the recombinant adenoviral vector of the invention comprises an expression cassette encoding a 3C protease or a 3CD protease of an enterovirus. The present invention also relates to a vaccine composition comprising the recombinant adenoviral vector as described. A method of inducing an immune response in a subject against enterovirus infection using the recombinant adenoviral vector and the vaccine composition is provided. Further provided is a method for producing virus like particles of an enterovirus by expressing the adenoviral vector as described herein in mammalian cells.

The present invention relates to a compound represented by formula (I) or a pharmaceutically acceptable salt or solvent compound thereof wherein M1 represents a 3-chymotrypsin-like protease inhibitor moiety; m2 represents an E3 ubiquitin ligase ligand part; l represents a linking group having a-(CH2CH2O) q (CH2) e (NH) f (O) g (CH2) hCO (CH2) i (CO) j-structure. The invention also relates to a preparation method of the compound and application of the compound in preparation of 3-chymotrypsin-like protease inhibitors and anti-coronavirus drugs.

The invention discloses a visible recombinant virus of human enterovirus 71 and an application thereof. In the invention, firstly a double-stranded DNA molecule is protected, wherein the double-stranded DNA molecule is obtained by inserting a coding gene labeled by a CCPGCC label into a 3C protease coding area of genomic DNA of the human enterovirus 71. The CCPGCC label is a polypeptide fragment represented as the sequence I in the sequence table. An expression kit, a recombinant vector, a transgenic cell line or a recombinant bacterium are all belongs to a scope of protection in the invention. The invention also protects a recombinant virus of the human enterovirus 71, wherein the coding DNA of a genomic RNA of the recombinant virus is represented as the sequence 4 in the sequence table. The visible recombinant virus of the human enterovirus 71 has an excellent application prospect in fundamental research.

Compounds of the formula: 1 where the formula variables are as defined in the disclosure, advantageously inhibit or block the biological activity of the picornaviral 3C protease. These compounds, as well as pharmaceutical compositions containing these compounds, are useful for treating patients or hosts infected with one or more picornaviruses, such as RVP. Intermediates and synthetic methods for preparing such compounds are also described.

The invention relates to a protease, particularly to a crystal structure of an enterovirus 71 3C protease and an application thereof. The enterovirus 71 3C protease is a wild protein without mutation, the crystal structure of the enterovirus 71 3C protease is composed of two domains, 14 beta-pleated sheets and 4 alpha-helixes form typical chymotrypsin sheet, and an active center is formed by a catalytic triad composed of His40, Glu71 and Cys147; and simultaneously, the crystal structure of the enterovirus 71 3C protease is provided with the same C2-D2 loop conformation with other 3C proteases of small ribonucleic acid (RNA) viruses. The crystal structure of the enterovirus 71 3C protease can be applied to preparation of small molecule inhibitors which can interact with specificity of the enterovirus 71 3C protease.