Peptide inhibitors of calcium oxalate monohydrate crystallization and uses thereof

Patent metadata

Field	Value
Publication number	US-10246489-B2
Application number	US-201715446253-A
Country	US
Kind code	B2
Filing date	Mar 1, 2017
Priority date	Jul 9, 2013
Publication date	Apr 2, 2019
Grant date	Apr 2, 2019

Abstract

In an embodiment, the present disclosure pertains to a composition for inhibiting calcium oxalate monohydrate crystal growth comprising at least one isolated polypeptide comprising a plurality of amino acids that bind the surface of the calcium oxalate monohydrate crystal; and a plurality of amino acids spacers, wherein the amino acid spacers are arranged in varying sequences between the plurality of amino acids that bind the surface of the calcium oxalate monohydrate crystal. In some embodiments, the present disclosure related to a method of controlling calcium oxalate monohydrate crystal growth in a subject in need thereof comprising administering to the subject therapeutically effective amount of the calcium oxalate monohydrate inhibiting polypeptide. In some embodiments, the present disclosure relates to a method of identifying calcium oxalate monohydrate inhibiting peptides. Such a method may comprise designing a peptide library of potential calcium oxalate inhibiting peptides; screening the peptide library for high efficacy inhibitor peptides for inhibition of calcium oxalate monohydrate crystallization; and conducting molecular characterization of the high efficacy inhibitor to determine specificity.

First claim

What is claimed is:

1. A method of identifying calcium oxalate monohydrate (COM) inhibiting peptides comprising the steps of:
(a) designing a peptide library of potential calcium oxalate inhibiting peptides, wherein the designing comprises:
selecting one or more amino acids that act as a binder to bind to a surface of a COM crystal,
selecting one or more amino acids that act as a spacer to minimize the steric hindrance of the amino acid binder to the surface of the COM crystal, and
synthesizing the potential calcium oxalate inhibiting peptides,
wherein the peptides comprise a plurality of regions comprising binder amino acids and a plurality regions comprising spacer amino acids, and
wherein the synthesizing comprises varying the number of spacer amino acids between the binder amino acids;

(b) screening the peptide library for high efficacy inhibitor peptides for inhibition of calcium oxalate monohydrate crystallization; and
(c) conducting molecular characterization of the high efficacy inhibitor peptides to determine specificity.

2. The method of claim 1 ,
wherein the one or more amino acids that act as the binder comprise at least one of glutamic acid (E), aspartic acid (D), and combinations thereof, and
wherein the one or more amino acids that act as the spacer comprise alanine (A).

3. The method of claim 1 , wherein the potential calcium oxalate inhibiting peptides are synthesized using solid-phase peptide synthesis.

4. The method of claim 1 , wherein the step of screening the peptide library for high efficacy inhibitor peptides comprises
mixing the test peptide with a supersaturated solution of calcium oxalate, and
measuring inhibition of growth of the COM crystal using in situ calcium ion-selective electrode measurement.

5. The method of claim 1 , wherein the step of molecular characterization of the high efficacy inhibitor peptides is performed utilizing Atomic Force Microscopy, Scanning Electron Microscopy, and Optical Microscopy.

6. The method of claim 1 , wherein the high efficacy inhibitor peptides comprise
at least one isolated polypeptide, and
at least one pharmaceutically acceptable carrier;
wherein the isolated polypeptide comprises an amino acid sequence selected from the group consisting of DDDAAAAADDDAAAAADD (SEQ ID NO: 1), AADAAAAADDAAAADAAA (SEQ ID NO: 2), ADAAADAADAADDAADAA (SEQ ID NO: 3), ADAADDAADAADDAAAAA (SEQ ID NO: 5), ADAAADDDAAADAAADDD (SEQ ID NO: 6), ADAAADDAAAAAAAADAA (SEQ ID NO: 7), ADAAADDAAADAAAADAA (SEQ ID NO: 8), ADAAADDAAADAAADDAA (SEQ ID NO: 9), ADAADAAADAADDAADAA (SEQ ID NO: 10), ADAADDAAAAAADAADAA (SEQ ID NO: 11), ADDAADAADAADDAADDA (SEQ ID NO: 12), AAEAAAAAEEAAAAEAAA (SEQ ID NO: 14), AEAAAEAAEAAEEAAEAA (SEQ ID NO: 15), AEAAAEEAAAEAAAAEAA (SEQ ID NO: 17), AEAAAEEAAAEAAAEEAA (SEQ ID NO: 18), AEAAEAAAEAAEEAAEAA (SEQ ID NO: 19), AEAAEEAAAAAAEAAEAA (SEQ ID NO: 20), AEEAAEAAEAAEEAAEEA (SEQ ID NO: 21), and AEEAEEAEEAEEAEEAEE (SEQ ID NO: 22), or consists of an amino acid sequence selected from the group consisting of ADAADAADAADAADAADA (SEQ ID NO: 4), ADADADADADADADADAD (SEQ ID NO: 13), AEAAEAAEAAEAAEAAEA (SEQ ID NO: 16), AEAEAEAEAEAEAEAEAE (SEQ ID NO: 23), and EEEEEEEEEEEEEEEEEE (SEQ ID NO: 24); and wherein the isolated polypeptide inhibits calcium oxalate monohydrate crystal growth.

Assignees

• Rimer Jeffrey D
• Karande Pankaj
• Univ Houston
• Rensselaer Polytech Inst

Inventors

• Rimer Jeffrey D
• Karande Pankaj

Classifications

• C40B30/04

  by measuring the ability to specifically bind a target molecule, e.g. antibody-antigen binding, receptor-ligand binding · CPC title

• G01N2800/345

  Urinary calculi · CPC title

• G01N2500/20

  cell-free systems · CPC title

• G01N33/6893

  related to diseases not provided for elsewhere · CPC title

• C07K7/08 Primary

  having 12 to 20 amino acids (gastrins C07K14/595; somatostatins C07K14/655; melanotropins C07K14/68) · CPC title