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Pharmaceutical composition or food composition, and method for assessing effect of active ingredient in vivo

a technology of active ingredients and compositions, applied in the field of pharmaceutical compositions or food compositions, can solve the problems of difficult early detection of declines in kidney function before onset of chronic kidney diseas

Pending Publication Date: 2022-03-31
KARYDO THERAPEUTIX INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The suppression of Oscar protein function in the composition effectively slows or improves declined kidney function and prevents CKD progression by reducing FGF23 expression, providing a method for evaluating the treatment's effectiveness through protein and mRNA measurement values.

Problems solved by technology

Thus, early detection of declined kidney function before onset of chronic kidney disease is difficult.

Method used

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  • Pharmaceutical composition or food composition, and method for assessing effect of active ingredient in vivo
  • Pharmaceutical composition or food composition, and method for assessing effect of active ingredient in vivo
  • Pharmaceutical composition or food composition, and method for assessing effect of active ingredient in vivo

Examples

Experimental program
Comparison scheme
Effect test

example 1

on of Soluble Human Oscar-Fc Fusion Protein

[0229]An expression plasmid containing a nucleotide sequence encoding a soluble human Oscar-Fc fusion protein was expressed in animal cells, thereby preparing the soluble human Oscar-Fc fusion protein shown in SEQ ID NO: 2. The preparation of the protein was outsourced to ProteinExpress Co., Ltd. (Chiba, Japan).

(SEQ ID NO: 2)MALVLILQLLTLWPLCHTDITPSVAIIVPPASYHPKPWLGAQPATVVTPGVNVTLRCRAPQPAWRFGLFKPGEIAPLLFRDVSSELAEFFLEEVTPAQGGSYRCCYRRPDWGPGVWSQPSDVLELLVTEELPRPSLVALPGPVVGPGANVSLRCAGRLRNMSFVLYREGVAAPLQYRHSAQPWADFTLLGARAPGTYSCYYHTPSAPYVLSQRSEVLVISWEGEGPEARPASDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK*

(The underline indicates a hinge region. The upstream from the hinge region indicates the sequence of human Oscar, and the downstream from the hinge region indicates t...

example 2

t of Administration of Soluble Human Oscar-Fc Fusion Protein to HP Model Mice

[0233]HP mice (mice ingested a diet with high phosphorus content) were obtained by feeding mice a diet with high phosphorus content. The soluble human Oscar-Fc fusion protein was administered to this model every week.

1. Phosphorus Overload

[0234]Mice ingested a diet with high phosphorus content (HP) were obtained by feeding mice (C57BL / 6N, 8 weeks old, male) a diet containing 2% inorganic phosphorus (TD.10662, OrientalBioService, Inc.) as a special phosphorus diet. Mice ingested a diet with low phosphorus content (LP) were fed a diet containing 0.35% inorganic phosphorus (TD.10662 modified type, OrientalBioService, Inc.).

2. Administration of Soluble Human Oscar-Fc Fusion Protein and Collection of Tissue

[0235]The soluble human Oscar-Fc fusion protein (10 mg / kg) was intraperitoneally administered weekly from the start date of the diet with high phosphorus content (week 0) to week 4, i.e., a total of five times...

experimental example i

Gene Mutant Mice

[0249]1. Construction of gRNA and Cas9 Expression Vector

[0250]gRNA sequences (Oscar-gRNA1 and Oscar-gRNA2) were designed, and oligo DNA encoding the gRNA sequences was synthesized using an Optimized CRISPR design tool (publicly available on the website of Massachusetts Institute of Technology, Zhang Lab (http: / / crispr.mit.edu / )). The Oscar gene target sequences contained in these sequences are the following sequences: ACAGCTGGAGTATCAGCGAC (SEQ ID NO: 20) in Oscar-gRNA1 and GCTCACAGAGAGTCGACAGC (SEQ ID NO: 21) in Oscar-gRNA2, which are present in exon 1 of the Oscar gene. These were individually inserted into pX330-U6-Chimeric_BB-CBh-hSpCas9, which is a Cas9 expression vector (pX330-Oscar-gRNA1 and pX330-Oscar-gRNA2). The nucleotide sequences of the gRNA insertion sites in the obtained vectors were determined to confirm that gRNA was inserted as designed. In addition, single-stranded oligodeoxynucleotides (ssODNs) were individually synthesized so as to include the res...

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PUM

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Abstract

An object of the present invention is to provide a pharmaceutical composition or food or drink composition comprising an active ingredient that suppresses functional expression of Oscar protein. Another object of the present invention is to provide a pharmaceutical composition or food composition for preventing or treating kidney disease. A further object of the present invention is to provide a pharmaceutical composition or food or drink composition that suppresses functional expression of Oscar in a living organism in order to suppress functional expression of FGF23. A still further object of the present invention is to provide a method for evaluating an effect, in the body, of an active ingredient that suppresses functional expression of Oscar protein. The above objects are achieved by at least one member selected from the group consisting of antagonists of the Oscar protein; genome editing systems that target Oscar gene; at least one RNA molecule selected from the group consisting of siRNA, shRNA, and miRNA that target Oscar mRNA, or vectors capable of expressing the RNA molecule; and antibodies that specifically bind to the Oscar protein and suppress function of the Oscar.

Description

[0001]A sequence listing in electronic (ASCII text file) format is filed with this application and incorporated herein by reference. The name of the ASCII text file is “P17-066US_Seq_Listing_210514.txt”; the file was created on May 18, 2021; the size of the file is 314 kilobytes (KB) or 322,454 bytes (b).TECHNICAL FIELD[0002]The present invention relates to a pharmaceutical composition or food composition comprising an active ingredient capable of suppressing functional expression of Oscar protein. The present invention also relates to a device, program, and method for evaluating an effect of the active ingredient in the body.BACKGROUND ART[0003]Diseases include those in a state that can be reversibly treated, and those in a state that cannot, i.e., those in an irreversible state. Early detection and treatment of abnormalities during a reversible state, or preventing such a state from occurring, is essential for health maintenance. Even in a reversible state, early detection of dise...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C07K14/705A61P13/12C12N9/22C12N15/11
CPCC07K14/70503A61P13/12A61K38/00C12N15/11C12N2310/20C12N9/22G01N33/6893C12Q1/6883G01N2333/75G01N2333/47G01N2800/347A61K31/7105A61K31/713A23L33/00G01N33/6812A01K67/0276A01K2217/075A01K2227/105A01K2267/035A01K2207/25A61K48/00C12N15/1138C12N2320/30C07K2319/30C12N2800/80C12Q2565/60
Inventor SATO, NARUTOKU
Owner KARYDO THERAPEUTIX INC
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