62 results about "PCSK9 Gene" patented technology

The present invention discloses a pharmaceutical composition comprising a circular RNA and a drug delivery carrier. Compared with traditional linear 1 * siRNA and annular 1 * siRNA, the number of repeated series connection of positive-sense strands is increased to include but not limited to two or more, it is accidentally found that the silence effect is remarkably enhanced, the expression level of the PCSK9 gene can be remarkably reduced, and degradation of mRNA of PCSK9 protein is mediated. The nano-particles are used for delivering oligonucleotide, so that the stability is improved, the immunogenicity is reduced, the effects of lowering cholesterol, reducing aortic plaque load and resisting atherosclerosis are improved, and the nano-particles are safe and free of obvious liver and kidney toxicity and have a wide application prospect in the aspect of preparing medicines for treating hypercholesterolemia and coronary heart disease.

Described are RNAi agents, compositions that include RNAi agents, and methods for dual inhibition of an Apolipoprotein C-III (APOC3) and Proprotein Convertase Subtilisin Kexin 9 (PCSK9) gene. The APOC3-PCSK9 RNAi agents disclosed herein inhibit the expression of an APOC3 and a PCSK9 gene. Pharmaceutical compositions that include one or more APOC3-PCSK9 RNAi agents, optionally with one or more additional therapeutics, are also described. Delivery of the described TSLP RNAi agents to hepatic cells, in vivo, provides for inhibition of APOC3 and / or PCSK9 gene expression, which can provide a therapeutic benefit to subjects, including human subjects, for the treatment of various diseases including hypertriglyceridemia and hypercholesterolemia.

Relating to the technical field of gene therapy, provided is an siRNA molecule for inhibiting PCSK9 expression, and use thereof. Particularly, an siRNA molecule for inhibiting PCSK9 expression (comprising a sense strand and an anti-sense strand that are complementary to form a double-stranded region) and a pharmaceutical composition comprising the same are disclosed. The siRNA molecule has a high silencing efficiency and a long-lasting silencing effect for PCSK9 gene, and is useful in the manufacture of products for inhibiting PCSK9 protein expression, products for reducing the concentration of low-density lipoprotein or low-density lipoprotein cholesterol in serum; and products for relieving the symptoms of PCSK9 gene-mediated diseases.

Provided is a pharmaceutical composition, comprising a circular RNA and a drug delivery carrier. It is surprisingly found that compared with conventional linear 1 x siRNAs and circular 1 x siRNAs, increasing the number of tandem repeats in the sense strand to two or more significantly enhances the silencing effect, significantly reduces the expression level of the PCSK9 gene, and mediates the degradation of the PCSK9 protein mRNA. The use of nano-particles for delivering oligonucleotides features improved stability, reduced immunogenicity, improved cholesterol-lowering, aortic plaque load-reducing and anti-atherosclerotic effects, good safety, no significant liver and kidney toxicity, and therefore good prospects in preparing medicaments for treating hypercholesterolemia and a coronary heart disease.

The present invention discloses an interfering RNA for inhibiting PCSK9 gene and use thereof. The interfering RNA comprises a nucleotide sequence set forth in any one or two or more of SEQ ID NOs: 1-40, 73-96. The interfering RNA of the present invention can better target and silence hepatic PCSK9 mRNA, reduce the protein level of PCSK9, enhance LDL-C metabolism, and reduce serum cholesterol, providing a solid technical foundation for the development of siRNA medicaments for the prevention, treatment, and symptom alleviation of PCSK9 gene-mediated diseases.

The invention relates to a siRNA for targeted regulation of PCSK9 gene expression and a use thereof. Experimental results show that some oligonucleotide sequences subject to alternate modification and specific template modification can significantly inhibit PCSK9 expression, and can be used for developing a drug for treating PCSK9-related diseases.

The invention discloses an interfering RNA (Ribonucleic Acid) for inhibiting a PCSK9 gene and application of the interfering RNA. The interfering RNA comprises any one or more than two nucleotide sequences as shown in SEQ ID NO: 1-40 and SEQ ID NO: 73-96. The interfering RNA can better target and silence mRNA of the liver PCSK9, reduce the protein level of the PCSK9, enhance LDL-C metabolism and reduce serum cholesterol, and a solid technical basis is provided for research and development of siRNA drugs for prevention and treatment of PCSK9 gene mediated diseases and symptom relief.

A method for simultaneously regulating the expression and / or activity of a dual-target gene. The method comprises: providing a complex or a nucleic acid encoding the complex, wherein the complex comprises a DNA binding domain and a gene expression modulator. For example, the method simultaneously regulates the expression and / or activity of PCSK9 gene and ANGPTL3 gene. For example, the method simultaneously regulates the expression and / or activity of APOC3 gene and the ANGPTL3 gene. For example, the method simultaneously regulates the expression and / or activity of the PCSK9 gene and the APOC3 gene.

The present disclosure relates to RNAi agents, e.g., double stranded RNAi agents such as small interfering RNA (siRNA) molecules, able to inhibit proprotein convertase subtilisin kexin 9 (PCSK9) gene expression. Also disclosed are pharmaceutical compositions that include PCSK9 RNAi agents and methods of use thereof. The PCSK9 RNAi agents disclosed herein may be conjugated to targeting ligands, including ligands that comprise N-acetyl-galactosamine, to facilitate the delivery to hepatocyte cells. Delivery of the PCSK9 RNAi agents in vivo provides for in vivo provides for inhibition of PCSK9 gene expression and thereby reduction of PCSK9 protein. The RNAi agents can be used in methods of treatment of diseases or disorders mediated at least in part by PCSK9 gene expression, including among others hypercholesterolemia, familial hypercholesterolemia including heterozygous familial hypercholesterolemia (HeFH) and homozygous familial hypercholesterolemia (HoFH), familial hypobetalipoproteinemia, hyperlipidemia, coronary artery disease, polygenic dyslipidemia, heart disease, cardiovascular disease (CVD) including clinical atherosclerotic cardiovascular disease (ASCVD).

Provided is an RNA inhibitor inhibiting PCSK9 gene expression, comprising an antisense strand, the antisense strand comprising a complementary region complementary to at least a portion of mRNA encoding PCSK9, and the length of the complementary region being 17-23 nucleotides; the antisense strand comprises any one of the following nucleotide sequences: SEQ ID NO.: 328-654, 1045-1107, 1125-1146 or 1149-1170, or a sequence differing therefrom by no more than 3 nucleotides. Also provided is a pharmaceutical composition, comprising the described RNA inhibitor inhibiting PCSK9 gene expression, and further comprising a delivery medium, and / or a pharmaceutically acceptable excipient and / or carrier and / or diluent.

The invention relates to an shRNA (short hairpin Ribonucleic Acid) interference sequence of a targeted silence PCSK9 gene and a construction method and lipid-lowering application of a recombinant adeno-associated virus vector of the shRNA interference sequence. Hyperlipidaemia is a metabolic disease characterized by abnormal rising of cholesterol and triglyceride levels in blood, and the design of lipid-lowering drugs is the focus of attention to improvement of hyperlipidaemia. Proprotein convertase subtilisin / kexin type 9 (PCSK9) can be combined with a low-density lipoprotein receptor (LDL-R) and degrade the LDL-R, so that accumulation of LDL-C in blood is further promoted, and hyperlipidemia is caused. Aiming at the key target PCSK9, a specific shRNA interference sequence is designed, and a recombinant adeno-associated virus vector (rAAV) carrying the sequence is constructed by an enzyme digestion-connection method. In-vitro experiments prove that the vector can remarkably reduce the expression level of PCSK9 protein, so that the cyclic utilization of a low-density lipoprotein receptor (LDL-R) is promoted, and the concentration of low-density lipoprotein cholesterol (LDL-C) in plasma is reduced. The rAAV vector provided by the invention has the characteristics of low production cost, high transfection efficiency, lasting action time and the like, and provides a new thought for gene therapy of hyperlipidemia.

A dual vector system for treating ornithine transcarbamylase deficiency is provided. The system includes ((a) a gene editing AAV comprising a first AAV rh79 capsid and a first vector genome comprising a 5′ ITR, a sequence encoding a meganuclease that targets PCSK9 under control of regulatory sequences that direct expression of the meganuclease in a target cell comprising a PCSK9 gene, and a 3′ ITR; and(b) a donor AAV vector comprising a second AAV capsid and a second AAV rh79 vector genome comprising: a 5′ITR, a 5′ homology directed recombination (HDR) arm, a transgene encoding ornithine transcarbamylase (OTC) and regulatory sequences that direct expression of the transgene in the target cell, a 3′ HDR arm, and a 3′ ITR.

Provided are a small interfering RNA (siRNA) used to inhibit the expression of proprote in convertase subtilisin / kexin type 9 (PCSK9) in a cell, a vector comprising nucleotides encoding same, and a cell, as well as a method using the siRNA, the vector or the cell for the treatment of diseases or symptoms associated with the expression of PCSK9 in a subject.

Provided are compositions that include compositions of galactosyl -conjugated lipid nanoparticles (LNPs) encapsulating one or more active agents. In some embodiments, the galactosyl- conjugated LNPs have a lipid component having D-Lin~MC3-DMA, ALC-0315 and. SM-102, cholesterol, DSPC and DOPE, and DMG-2000-PEG. In some embodiments, the GalNAc-conjugated LNP has one or more galactosyl moieties bioconjugated to cholesterol present with a lipid component of the GalNAc -conjugated LNP. Also provided are methods for treating and / or preventing diseases, disorders, and / or conditions associated, with undesirable PCSK9 gene expression, optionally a cardiovascular disease, disorder, or condition, including but not limited, to atherosclerosis and / or thrombosis, and sepsis, septic shock, cytokine storm, or sequelae thereof.

The present disclosure relates to siRNA and its use for targeting control of PCSK9 gene expression.Experimental results show that oligonucleotide sequences with some alternating modifications and specific template modifications can significantly inhibit PCSK9 expression, and can be used to develop drugs for the treatment of PCSK9-related diseases.

Provided herein are compositions, systems, and methods for modifying a human APOC3 gene, PCSK9 gene, or ANGPTL3 gene. Systems, compositions, and methods may comprise a CRISPR-associated (Cas) protein or uses thereof. Systems, compositions, and methods of the present disclosure may be useful for treatment of APOC3 associated conditions, including familial chylomicronemia syndrome (FCS) and severe hypertriglyceridemia (SHTG).

The invention relates to a siRNA for targeted regulation of PCSK9 gene expression and a use thereof. Experimental results show that some oligonucleotide sequences subject to alternate modification and specific template modification can significantly inhibit PCSK9 expression, and can be used for developing a drug for treating PCSK9-related diseases.

Disclosed herein are methods of disease treatments by administering to a subject in need thereof a pharmaceutical composition comprising gene editing compositions. The methods disclosed herein can provide durable in vivo editing of PCSK9. More particularly, the compositions disclosed herein are capable of in vivo editing PCSK9 gene in humans and reducing PCSK9 protein and LDL-C for a potentially transformative treatment of cardiovascular disease, the leading cause of death in the United States and world-wide. Various aspects of the compositions, use and preparation are disclosed, including clinically supported therapeutically effective and safe human dosing of the compositions.

The present application relates to siRNA, siRNA conjugates, pharmaceutical compositions comprising the same, and uses thereof for inhibiting the expression of PCSK9 gene. Each nucleotide in the siRNA is independently modified or unmodified, and the siRNA comprises a sense strand and an antisense strand. The siRNA, conjugates thereof, and pharmaceutical compositions thereof can be effective in treating and / or preventing diseases associated with overexpression of PCSK9 gene.

This invention discloses a circular RNA agent targeting PCSK9 and its applications, relating to the field of biomedical technology. The circular RNA agent is a single-stranded closed-circular oligonucleotide containing an antisense sequence complementary to the transcription of the PCSK9 gene. It lacks free 5' and 3' ends, and does not contain overhangs, hairpin structures, or double-stranded regions. The molecules are linked by 3'-5' phosphodiester bonds. This circular RNA agent can be modified with GalNAc or prepared into pharmaceutical compositions for the treatment of diseases or conditions related to abnormal PCSK9 gene expression or activity. The circular RNA agent provided by this invention exhibits high stability, a long half-life, low immunogenicity, low modification toxicity, and high target gene inhibitory activity. It can significantly prolong the dosing interval, improve patient compliance, and provide a safer, longer-acting drug option for lipid-lowering therapy.

The invention discloses a composition for gene editing and application thereof. The composition comprises i) a CRISPR / Cas gene editing element and a delivery vector thereof, and ii) a recombinant adeno-associated virus (AAV) vector carrying a target gene, and the target gene comprises at least one selected from the following genes: a blood coagulation factor VIII (F8) gene, an LDLR gene, a TTR gene, a PAH gene, an F9 gene, a PCSK9 gene or an SERPINC1 gene. The composition for gene editing disclosed by the invention can realize efficient gene integration and expression, reduce immune response and increase safety, and is suitable for patients of all ages, so that effective treatment choices are provided for different people.

A dual component system for treating a genetic disorder is provided. The system includes (a) a gene editing vector comprising an expression cassette comprising a nucleic acid sequence encoding a nuclease and regulatory sequences that direct expression of the nuclease in a target cell comprising a PCSK9 gene; and (b) a donor vector comprising a nucleic acid sequence encoding an exogenous product for expression from the PCSK9 locus, wherein the inserted nucleic acid sequence does not encode PCSK9, wherein the system further comprises sequences that direct the nuclease to specifically targets the native PCSK9 gene locus; and wherein the native PCSK9 in the target cell is optionally ablated or reduced post-dosing with the dual component system.

A method for preparing enhanced CAR-T-cells based on interfering or knocking out the human PCSK9 gene and an application thereof are provided. The method includes performing the RNA interference (RNAi) for the human PCSK9 gene by the artificial microRNA (miRNA) to prepare the enhanced CAR-T or preparing the enhanced CAR-T by CRISPR-Cas9 gene-editing technology by using sgRNA to knock out the human PCSK9 gene. The method uses PCSK9 gene knockdown element or CRISPR / Cas9 technology to specifically knock down or knock out the expression of PCSK9 in CAR-T-cells, which effectively enhances the therapeutic effect of CAR-T-cells on malignant solid tumors, and can enhance the tumor therapeutic effect of immune checkpoint blockade therapy.

This invention relates to the field of gene editing technology, and in particular to an sgRNA, a gene editing system, and their uses. The sgRNA comprises a target sequence targeting the PCSK9 gene or a target sequence targeting the APOC3 gene. The nucleotide sequence encoding the target sequence targeting the PCSK9 gene comprises a sequence as shown in any one of SEQ ID Nos. 1-22; the nucleotide sequence encoding the target sequence targeting the APOC3 gene comprises a sequence as shown in any one of SEQ ID Nos. 23-36. Through systematic screening and experimental verification, this invention has obtained sgRNAs capable of efficiently and specifically targeting both the PCSK9 and APOC3 genes. Based on the screened sgRNAs, this invention has developed a dual-target gene editing system capable of simultaneously targeting both the PCSK9 and APOC3 genes. This provides a reliable core component for developing gene-editing-based therapies for lipid metabolism diseases, and has significant clinical significance and application prospects.

Provided in the present invention is a preparation method for a siRNA for inhibiting PCSK9 gene expression. The siRNA in the preparation method is a double-stranded RNA consisting of a sense strand and an antisense strand which are complementarily paired. The preparation method comprises: mixing a sense strand substrate, an antisense strand substrate and an RNA ligase, using the RNA ligase for catalyzing the sense strand substrate and the antisense strand substrate to be connected by means of a phosphodiester bond, and obtaining a sense strand and an antisense strand, so as to obtain the siRNA. The sense strand substrate can form the sense strand, and the antisense strand substrate can form the antisense strand. The sense strand is a nucleic acid sequence as shown in SEQ ID NO: 45, and the antisense strand is a nucleic acid sequence as shown as SEQ ID NO: 46. The solution solves the problem in the prior art of low purity of the siRNA, and is applicable in the field of medicine biosynthesis.

The invention belongs to the field of biological medicine, and relates to siRNA, a conjugate thereof and application of the siRNA and the conjugate in related diseases such as blood fat. The siRNA comprises a positive-sense strand and an antisense strand, the positive-sense strand comprises a nucleotide sequence I, and the antisense strand comprises a nucleotide sequence II; each nucleotide in the nucleotide sequence I and the nucleotide sequence II is modified or unmodified nucleotide; the nucleotide sequence I and the nucleotide sequence II are at least partially reversely complementary to form a double-stranded region; the nucleotide sequence I is basically consistent with the first nucleotide sequence, and the first nucleotide sequence is a nucleotide sequence with the length of at least 19 nucleotides in mRNA expressed by the PCSK9 gene. According to the present invention, the siRNA and the conjugate thereof can specifically target the liver, can be complementarily paired with the liver PCSK9 mRNA sequence, and can induce the PCSK9 mRNA degradation, such that the synthesis of the PCSK9 in the liver can be inhibited, the PCSK9 serum level can be durably and significantly reduced, and the related diseases caused by excessive PCSK9 can be effectively prevented and / or treated.

The present application relates to an epigenetic editing agent, which contains a transcription activator-like effector (TALE) domain, at least one epigenetic modification domain, and at least one transcriptional regulatory domain, wherein the TALE domain can specifically bind to a target nucleotide sequence on the PCSK9 gene and / or on a regulatory element of the PCSK9 gene.

A composition for editing the PCSK9 gene comprises: (i) an mRNA encoding a protein comprising a programmable DNA binding domain; and (ii) a guide polynucleotide, wherein the guide polynucleotide compr

This article provides, in particular, dsRNA compositions targeting PCSK9, methods for inhibiting PCSK9 gene expression, and methods for treating one or more diseases associated with PCSK9 gene expression.

This disclosure relates to modified guide RNAs having improved in vitro and in vivo activity in genome editing methods. In some embodiments, the present disclosure provides compositions and methods for modifying a PCSK9 gene. In some aspects, the present disclosure provides a guide RNA, compositions thereof, and pharmaceutical compositions comprising a guide RNA or a composition as described herein. In some aspects, the present disclosure also provides uses and methods of using a guide RNA, a composition thereof, or a pharmaceutical composition as described herein, for inducing a double-strand break or a single-strand break in a PCSK9 gene, for reducing expression of a PCSK9 gene in a cell or subject, and for treating a patient having or at risk of having a PCSK9-related disease or condition.