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Polypeptide micromolecule and application thereof in preparation of medicine for preventing and treating bladder overactivity

An over-bladder, small molecule technology for biomedical applications

Active Publication Date: 2014-05-14
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] There is currently no information on the inhibition of P2X by small peptide molecules 3 Receptor activity to relieve overactive bladder

Method used

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  • Polypeptide micromolecule and application thereof in preparation of medicine for preventing and treating bladder overactivity
  • Polypeptide micromolecule and application thereof in preparation of medicine for preventing and treating bladder overactivity
  • Polypeptide micromolecule and application thereof in preparation of medicine for preventing and treating bladder overactivity

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Embodiment 1: The preparation method of polypeptide small molecule

[0018] 1. The amino acid sequence of the 14 peptide is shown in SEQ ID NO: 1:

[0019] Met-Glu-Val-Leu-Pro-Lys-Ala-Leu-Glu-Val-Asp-Glu-Arg-Ser

[0020] 2. The amino acid sequence of TAT penetrating peptide is shown in SEQ ID NO:2:

[0021] Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg

[0022] In order to promote the entry of polypeptide molecules into the cytoplasm, a TAT membrane-penetrating peptide was fused to the C-terminus of No. 1 polypeptide molecule (Schwarze SR, Ho A, Vocero-Akbani A, Dowdy SF. In vivo protein transduction: delivery of a biologically active protein into the mouse. Science. 1999 Sep 3;285(5433):1569-72.).

[0023] The above polypeptide molecules were synthesized by Shanghai Keyept Biotechnology Co., Ltd., with a purity of more than 95%. Example 2: Small polypeptide molecules to P2X 3 Inhibition of receptor activity

Embodiment 2

[0023] The above polypeptide molecules were synthesized by Shanghai Keyept Biotechnology Co., Ltd., with a purity of more than 95%. Example 2: Small polypeptide molecules to P2X 3 Inhibition of receptor activity

[0024] 1. Experimental method

[0025] pcDNA3.1-rP2X 3 Plasmid (pcDNA3.1 vector was purchased from Invitrogen, rP2X 3 Obtained from rat dorsal root ganglion tissue by RT-PCR method, and cloned into pcDNA3.1 vector) and pEGFP-N2 plasmid (purchased from Clontech Company) co-transfected into HEK293T cells (purchased from Shanghai Institute of Biological Sciences, Chinese Academy of Sciences Hospital Cell Resource Center), after 24 to 36 hours, the cells were incubated with polypeptides for 1 hour. Under the fluorescence microscope, the cells with EGFP green fluorescence are considered to express P2X at the same time 3 recipient cells.

[0026] P2X expressed by these cells was recorded by the patch clamp method ("Patch Clamp Technology and Its Application", edited ...

Embodiment 3

[0036] Example 3: Animal experiment of small polypeptide molecules used in the treatment of overactive bladder

[0037]Cyclophosphamide-induced cystitis is a commonly used model to study overactive bladder (Everaerts W, Zhen X, Ghosh D, Vriens J, Gevaert T, Gilbert JP, Hayward NJ, McNamara CR, Xue F, Moran MM, Strassmaier T, Uykal E, Owsianik G, Vennekens R, De Ridder D, Nilius B, Fanger CM, Voets T. Inhibition of the cation channel TRPV4 improves bladder function in mice and rats with cyclophosphamide-induced cystitis. Proc Natl Acad Sci USA. 2010 Nov 2 ;107(44):19084-9.; Ochodnicky P, Cruz CD, Yoshimura N, Cruz F. Neurotrophins as regulators of urinary bladder function. Nat Rev Urol.2012 Nov;9(11):628-37.; Neuhaus J , Schwalenberg T. Intravesical treatments of bladder pain syndrome / interstitial cystitis. Nat Rev Urol. 2012 Dec;9(12):707-20.). Therefore, the present invention uses this model to verify the therapeutic effect of the polypeptide on overactive bladder.

[0038]...

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Abstract

The invention relates to the technical field of biological medicines. The bladder overactivity is presented clinically with frequent micturition and urgent urination, and also appears in neurogenic bladder, overactive bladder, bladder outlet obstruction, interstitial cystitis and other diseases. A polypeptide micromolecule capable of inhibiting the activity of a P2X3 acceptor is provided and has an amino acid sequence as shown in SEQ ID NO.: 1. The invention also provides application of the polypeptide micromolecule in preparation of a medicine for preventing and treating bladder overactivity.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a small polypeptide molecule capable of reducing the excitability of peripheral nerve endings and relieving overactive bladder and its application in the preparation of drugs for preventing and treating overactive bladder. Background technique [0002] The clinical manifestations of overactive bladder are frequent urination and urgency, which are manifested in various diseases such as neurogenic bladder, overactive bladder, bladder outlet obstruction, and interstitial cystitis. In the United States, about 7.9 million women over the age of 18 suffer from such diseases (Neuhaus J, Schwalenberg T. Intravesical treatments of bladder pain syndrome / interstitial cystitis. Nat Rev Urol. 2012Dec;9(12):707-20.) . In Asia, more than 22.9% of people over the age of 30 have overactive bladder (Yoo ES, Kim BS, Kim DY, Oh SJ, Kim JC. The impact of overactive bladder on health-related quali...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/08A61K38/10A61P13/00A61P13/10
Inventor 何成高霄飞冯吉锋王伟
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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