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Anti-periostin antibody and pharmaceutical composition for preventing or treating periostin-related disease containing the same

A technology of periostin and composition, applied in drug combination, bone disease, antibody, etc., can solve problems such as irrelevance, inability to guarantee the quality of life, and inability to fully ensure the prognosis of patients with heart failure.

Inactive Publication Date: 2009-03-18
OSAKA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is still a problem that patients with heart failure are restricted in their daily life such as being unable to exercise vigorously, and the quality of life cannot be ensured, so that the life prognosis of heart failure patients cannot be fully ensured, so it is desired to develop a drug that can improve the quality of life and long-term life prognosis Potent new heart failure drug
However, no antibody related to the structure of the region involved in the cell adhesion activity of periostin has been reported, and there is no report on the relationship between the cell adhesion activity of periostin and diseases such as heart failure

Method used

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  • Anti-periostin antibody and pharmaceutical composition for preventing or treating periostin-related disease containing the same
  • Anti-periostin antibody and pharmaceutical composition for preventing or treating periostin-related disease containing the same
  • Anti-periostin antibody and pharmaceutical composition for preventing or treating periostin-related disease containing the same

Examples

Experimental program
Comparison scheme
Effect test

manufacture example 1

[0137] [Manufacturing Example 1] Search for periostin by exclusion method

[0138] 1-1 Preparation of heart failure model rats and extraction of left ventricle samples

[0139] From the age of 6 weeks, male Dah1 salt-sensitive rats (Dahl-S) (experimental materials in clear water) were fed with high-salt food containing 8% salt. The left ventricles of three mice were removed.

[0140] [51] 1-2 mRNA preparation

[0141] Total RNA was prepared from about 500 mg of the above left ventricle using ISOGEN (Nippon Gene) according to the method described in the manual. Next, mRNA was purified from about 400 μg of total RNA from each of three rats in the cardiac hypertrophy and heart failure phases using Fast Track 2.0 Kit (Invitrogen) according to the method described in the manual, and about 3 μg of mRNA were recovered.

[0142] [52] 1-3 cDNA subtraction

[0143] The cDNA subtraction was carried out by using the PCR-Select cDNA subtraction kit (Clontech Company) according to the m...

Embodiment 1

[0178] [75] Synthesis of Rat Exon-17 Peptide Chain and Preparation of Polyclonal Antibody

[0179] If the PN-1 gene is highly expressed in the heart of normal SD rats, heart dilation will be caused, and if the antisense oligonucleotide of rat periostin is administered in the heart of Dahl heart failure model rats, then Survival was improved. In addition, unlike the reported PN-2, it is clear that there is no cell adhesion in rat PN-1. Based on the comparison of the respective sequences, the specific structure in rat PN-1 was identified as is the exon-17 sequence. 10 mg of a peptide having a Cys residue added to the N-terminal of the amino acid sequence composed of exon-17 was chemically synthesized with a purity of 80% or higher. The peptide was combined with 6 mg of KLH as a carrier protein, and rabbits (Kb1:JW) were immunized again. As for the immunization method, FCA (Freund's complete adjuvant) was used for the first immunization, and FIA (Freund's incomplete adjuvant) w...

Embodiment 2

[0181] [76] In vitro study of the anti-cell adhesion activity of rat PN-1

[0182]By comparing with documented methods (Ruwhof C, van Wamel AE, Egas JM, van der Laarse A. Mol Cell Biochem. 2000 May; 208(1-2):89-98, Ashizawa N, Graf K, Do YS, Nunohiro T, Giachelli CM, Meehan WP, Tuan TL, Hsueh WA. J Clin Invest. 1996 November 15; 98(10): 2218-27) obtained cardiac fibroblasts of rats by the same method. Specifically, 20 SD rats aged 1-2 days after birth were anesthetized with ether, and their chests were disinfected with ethanol. Remove the heart, put it into a dish filled with PBS(-), cut the heart according to the cross-section, the blood flows out, wash with PBS(-) 3 times, remove the PBS(-) as much as possible, and cut it finely with scissors. Then, mix PBS(-) with collagenase / trypsin at a ratio of 1:1, shake at 37°C for 15 minutes, and perform petting to separate the cells as much as possible. Then, put the platinum sieve into a centrifuge tube to filter the cell separati...

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Abstract

The invention provides an antibody against periostin having an anti-cell adhesion activity, particularly the anti-periostin antibody having an ability to neutralize an anti-cell adhesion action, and a preventive or therapeutic agent for a periostin-related disease using the antibody. It is also intended to provide detection and determination methods for the periostin in a sample using the antibody and a diagnostic method for a periostin-related disease by determining the amount of the periostin by the method.

Description

[0001] Detailed Description of the Invention [01] Technical field [0002] [02] The present invention relates to an antibody against periostin having an anti-cell adhesion effect, particularly an anti-periostin antibody having an ability to neutralize the anti-cell adhesion effect. More specifically, an antibody useful in the prevention or treatment of periostin-related diseases such as heart failure or in the diagnosis of these diseases, which is specifically expressed in the interstitium of tissue reconstruction represented by cardiac hypertrophy tissue, has Anti-periostin antibody against splice variant of periostin for cell adhesion. Background technique [0003] [03] Chronic heart failure is a disease in which the heart cannot pump enough blood to various organs due to the decrease of myocardial contractility. In the past, cardiotonic agents such as digitalis preparations that increase myocardial contractility have been used in its treatment. However, since these drug...

Claims

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Application Information

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IPC IPC(8): C07K16/18A61K39/395A61P1/16A61P1/18A61P9/00A61P9/04A61P11/00A61P13/12A61P19/04A61P19/10A61P29/00A61P35/00C07K16/46G01N33/53C12P21/08
CPCG01N2800/32C07K16/22G01N33/574A61K2039/505C07K16/18C07K2316/96A61P1/16A61P1/18A61P9/00A61P9/04A61P9/10A61P11/00A61P13/12A61P19/02A61P19/04A61P19/10A61P25/28A61P29/00A61P35/00A61P43/00C07K2317/76A61K39/395C07K16/46G01N33/53
Inventor 谷山义明森下龙一葛城鸣门
Owner OSAKA UNIV
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