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Stable amorphous calcium carbonate comprising synthetic phosphorylated peptides

a technology of phosphorylated peptides and calcium carbonate, which is applied in the direction of phosphorous compound active ingredients, immunological disorders, metabolism disorders, etc., to achieve the effect of preventing or reducing calcium carbonate crystallization

Inactive Publication Date: 2010-09-02
AMORPHICAL LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0004]Other objects and advantages of present invention will appear as description proceeds.

Problems solved by technology

The formation of amorphous calcium carbonate in the living bodies of, for example, crayfish is rather intriguing, since amorphous minerals are usually thermodynamically unstable.

Method used

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  • Stable amorphous calcium carbonate comprising synthetic phosphorylated peptides
  • Stable amorphous calcium carbonate comprising synthetic phosphorylated peptides
  • Stable amorphous calcium carbonate comprising synthetic phosphorylated peptides

Examples

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example 1

[0083]Gastroliths of Cherax quadricarinatus were prepared as described [WO 2005 / 115414]. SDS-PAGE separation of soluble proteins from late premolt gastroliths revealed the presence of at least 6 prominent distinct proteins (FIG. 1A left) with the most abundant being at the size of approximately 65 kDa (gastrolith protein 65, GAP65). Further purification of GAP65 from the entire gastrolith soluble proteins content was performed using DEAE chromatography HPLC with NaCl gradient of up to 1M. GAP65 elution began at 300 mM NaCl but continued mainly at 600 mM (fraction 17). The GAP65 enriched fraction 17 was analyzed by SDS-PAGE and stained with Coomassie (non specific protein staining), “stains all” (negatively charged protein staining), and “pas” (glycoprotein staining), as shown in FIG. 1A right. These staining suggest that GAP65 is the primary protein in this enriched fraction and it is a negatively charged glycoprotein. Trypsin digestion of GAP65 followed by separation using nanospra...

example 2

[0084]Specific expression of GAP65 was tested in premolt crayfish in several target tissues by means of RT-PCR (FIG. 3A). The expression of GAP65 was detected in the gastrolith epithelial disc and in the sub-epidermal tissue, both are cuticle related tissues. Expression of GAP65 was not detected in the hepatopancreas, stomach wall, and sperm duct. Localization of GAP65 expression in the gastrolith disc of induced premolt and intact intermolt crayfish by in situ hybridization is presented in FIG. 3B. Left panel represents hematoxylin and eosin staining of the gastrolith disc, middle panel is the control sense probe where no expression is detected. The two right panels represent the anti-sense probe with the last being an enlargement of a specific area. The anti-sense probe reveals that the expression of GAP65 can be detected only in the columnar epithelial cells of the gastrolith disc of an induced crayfish, whereas, in intact intermolt crayfish this expression was not detected.

example 3

[0085]Relative GAP65 transcript levels in the gastrolith epithelial disc following silencing using GAP65 dsRNA were measured using realtime RT-PCR and presented in FIG. 4. GAP65 levels were evaluated in crayfish injected with ecdysone and GAP65 dsRNA, ecdysone and dsRNA carrier, ecdysone and C. quadricarinatus vitellogenin (CqVg) dsRNA, and a control injected with both carriers. CqVg, an hepatopancreatic specific gene found mostly in reproductive females, served as a control for sequence specific silencing. Transcript levels of crayfish injected with ecdysone and GAP65 dsRNA were significantly lower than the levels found in the ecdysone and dsRNA carrier injected. In the crayfish injected with ecdysone and CqVg dsRNA, GAP65 transcript levels were similar to the levels detected in the ecdysone and dsRNA carrier injected group. In the control carriers injected crayfish GAP65 transcript levels were higher than the levels found in the ecdysone and GAP65 dsRNA injected crayfish but lower...

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Abstract

Provided are compositions containing amorphous calcium carbonate (ACC), and at least one phosphorylated peptide which stabilizes the amorphous form of said calcium carbonate. Particularly, the peptide can be selected from crustacean proteins, also provided by the invention, namely GAP65, GAP22, GAP21, and GAP12 (also indicated herein as GAP10). The compositions are useful in pharmaceutical and nutraceutical formulations.

Description

FIELD OF THE INVENTION[0001]The present invention relates to compositions comprising amorphous calcium carbonate, and to methods of preparing same, and further to compositions comprising phosphorylated amino acids or peptides. Particularly, said peptides are selected from crustacean proteins, including GAP65, GAP22, GAP21, and GAP12 (also referred to herein interchangeably as “GAP10”). Pharmaceutical and nutraceutical compositions comprising amorphous calcium carbonate and phosphorylated amino acids or peptides are provided.BACKGROUND OF THE INVENTION[0002]Calcium plays one of the central roles in the signal transduction, and further it is an important structural element in biological systems. From protozoa to vertebrata, deposited calcium salts helps to keep rigid bodily shapes of animals, calcium phosphate being the main component of endoskeletons in the vertebrates and calcium carbonate of exoskeletons in the invertebrates. Calcified exoskeletons with calcium carbonate minerals a...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61K33/10C07K14/00C07H21/00A61P19/10A23L1/30A23L33/00
CPCA61K31/66A61K33/10A61K38/17C07K14/43509A61K2300/00A61K38/1767A61P1/02A61P11/00A61P15/00A61P19/08A61P19/10A61P21/00A61P25/00A61P25/04A61P25/16A61P25/28A61P29/00A61P3/00A61P3/02A61P3/14A61P35/00A61P37/00A61P37/02A61P9/00A61K9/0053
Inventor BENTOV, SHMUELSAGI, AMIRBERMAN, AMIRSHECHTER, ASSAF
Owner AMORPHICAL LTD
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