# US 10,246,489 B2 · Peptide inhibitors of calcium oxalate monohydrate crystallization and uses thereof

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- Number: US 10,246,489 B2
- Document ID: US10246489B2
- Status: Granted
- Kind: B2
- Application number: 15/446,253
- Filed: 2017-03-01
- Published: 2017-10-05
- Granted: 2019-04-02
- Earliest claimed: 2013-07-09
- Assignees: University of Houston; Rensselaer Polytechnic Institute
- Applicants: Jeffrey D. Rimer; Pankaj Karande
- Inventors: Jeffrey D. Rimer; Pankaj Karande
- Web page: https://patentgrep.com/patent/US10246489B2

## CPC

- C07K 7/08: having 12 to 20 amino acids (gastrins C07K14/595; somatostatins C07K14/655; melanotropins C07K14/68)
- A61P 13/00: Drugs for disorders of the urinary system
- A61P 13/12: of the kidneys
- G01N 33/53: Immunoassay; Biospecific binding assay; Materials therefor
- G01N 33/6893: {related to diseases not provided for elsewhere}
- A61K 38/00: Medicinal preparations containing peptides (peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00)
- C40B 30/04: by measuring the ability to specifically bind a target molecule, e.g. antibody-antigen binding, receptor-ligand binding
- G01N 2500/04: Screening involving studying the effect of compounds C directly on molecule A (e.g. C are potential ligands for a receptor A, or potential substrates for an enzyme A)
- G01N 2500/20: cell-free systems
- G01N 2800/345: Urinary calculi
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- C40B: COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES (in silico combinatorial libraries of nucleic acids, proteins or peptides G16B35/00; in silico combinatorial chemistry G16C20/60)

## 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.

## Claims

### Claim 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.

### Claim 2 (depends on 1)

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).

### Claim 3 (depends on 1)

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

### Claim 4 (depends on 1)

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.

### Claim 5 (depends on 1)

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.

### Claim 6 (depends on 1)

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.