The present embodiments provide methods, compounds, and compositions useful for inhibiting PCSK9 expression, which may be useful for treating, preventing, or ameliorating a disease associated with PCSK9.

The present invention provides compositions comprising an isolated or purified therapeutically effective hPCSK9 polypeptide derived from the hPCSK9 catalytic domain, and their use in methods of treating hypercholesterolemia.

The invention provides a method for the large-scale preparation of small peptides using recombinant DNA technology. Overexpression of small peptides, such as liraglutide precursor, as concatemers, improves the overall efficiency of the process due to increased yields per batch of the biologically active peptide. Digestion of these concatemers by combinations of specific enzymes yields the desired peptide monomer in large quantities. More particularly, the invention relates to the production of recombinant peptide precursor of liraglutide

The present invention provides novel polynucleotides encoding PCSK9b and PCSK9c polypeptides, fragments and homologues thereof. Also provided are vectors, host cells, antibodies, and recombinant and synthetic methods for producing said polypeptides. The invention further relates to diagnostic and therapeutic methods for applying these novel PCSK9b and PCSK9c polypeptides to the diagnosis, treatment, and/or prevention of various diseases and/or disorders related to these polypeptides. The invention further relates to screening methods for identifying agonists and antagonists of the polynucleotides and polypeptides of the present invention.

The present embodiments provide methods, compounds, and compositions useful for inhibiting PCSK9 expression, which may be useful for treating, preventing, or ameliorating a disease associated with PCSK9.

Disclosed are compounds of Formula I, or a salt thereof:

##STR00001##

where A, B, D, X, R.sup.1, R.sup.2 and R.sup.8 are as defined herein, which compounds have properties for antagonizing PCSK9. Also described are pharmaceutical formulations comprising the compounds of Formula I or their salts, and methods of treating cardiovascular disease and conditions related to PCSK9 activity, e.g. atherosclerosis, hypercholesterolemia, coronary heart disease, metabolic syndrome, acute coronary syndrome, or related cardiovascular disease and cardiometabolic conditions.

The present invention relates to a method for modulating stem cells proliferation or differentiation comprising a step of contacting said stem cells with an effective amount of an activator or inhibitor of a proprotein convertase subtilisin kexin 9 (PCSK9). Inventors performed a global transcriptomic analyses in hiPSCs and showed that PCSK9 inhibition by shRNA and the intracellular PCSK9-R104C/V114A mutation negatively regulate the NODAL signaling pathway and its targets. This regulation was manifested in drastic reduction P-SMAD2/total SMAD2 protein level. This was accompanied by reduced proliferation rate where hiPSC-shPCSK9 and hiPSC-R104C/V114A demanded >1.3-fold more time to double compared to their control counterparts. They showed that PCSK9 was regulating this signaling pathway through direct physical interaction with DACT2, an intracellular attenuator of NODAL receptor and favoring its protein degradation. Thus, these findings allow to understand the differentiation and proliferation of cells.

Provided herein are double-stranded nucleic acid inhibitor molecules having a sense strand with a stem loop structure and an antisense strand, where the stem portion of the stem loop structure contains one or more T.sub.m-increasing nucleotides. Also provided are methods and compositions for reducing target gene expression and methods and compositions for treating a disease of interest.

The present invention relates to a vaccine capable to induce the formation of antibodies directed to PCSK9 in vivo.

The invention relates to various PCSK9 RNAi constructs with gene silencing activities, and uses thereof. The construct has a double-stranded region of 19-49 nucleotides, preferably 25, 26, or 27 nucleotides, and preferably blunt-ended. The construct has selective minimal modifications to confer an optimal balance of biological activity, toxicity, stability, and target gene specificity. The sense strand may be modified such that the construct is not cleaved by Dicer or other RNAse III, and the entire length of the antisense strand is loaded into RISC In addition, the antisense strand may also be modified by 2-O-methyl groups at the 2nd 5-end nucleotide to greatly reduce off-target silencing. The constructs of the invention largely avoid the interferon response and sequence-independent apoptosis in mammalian cells, exhibits better serum stability, and enhanced target specificity.