Novel methods of use of hsp70 for increased performance or treatment of hsp70 related disorders

a technology of hsp70 and hsp70, which is applied in the direction of peptide/protein ingredients, drug compositions, muscular disorders, etc., can solve the problems of limited approach, limited key therapeutic approaches and care standards, and the tens of billions of dollars associated with treatment and care of musculoskeletal diseases. achieve the effect of fatigue and increase performan

Inactive Publication Date: 2013-04-11
ALTERNATIVE INNOVATIVE TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0013]In another aspect, the invention provides a method to increase performance (e.g., endurance), alleviate a fatigue syndrome, or treat muscle damage or muscle degeneration comprising administering to the mammal a compound of the invention, e.g., a therapeutically effective amount of HSP70, e.g., human heat shock protein 70, such that performance (e.g., endurance) is increased, the fatigue syndrome is alleviated, or the muscle damage or muscle degeneration is treated in the mammal.

Problems solved by technology

Annual costs associated with treatment and care of musculoskeletal disease is in the tens of billions of dollars.
Current key therapeutic approaches and standards of care are typically limited to physiotherapeutic rehabilitation, pain management and anti-inflammatory medications.
However, this approach is limited by poor integration of the transplanted cells and their low survival.
It is notable that most of the work was done with a vascular endothelial growth factor VEGF, a well-known proangiogenic regulator and thus it was challenging to separate angiogenic contributions from direct effects on muscle development and evolution of the related progenitor cells population.
It has been shown that tissue injury, whether caused by surgery, trauma or disease, results in the induction of heat shock / stress proteins.
Although evidence has suggested the role of HSP70 in certain indications, current treatments that have adopted strategies to control in vivo HSP70 production have not met the need in this arena.

Method used

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  • Novel methods of use of hsp70 for increased performance or treatment of hsp70 related disorders
  • Novel methods of use of hsp70 for increased performance or treatment of hsp70 related disorders
  • Novel methods of use of hsp70 for increased performance or treatment of hsp70 related disorders

Examples

Experimental program
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Effect test

example 1

Fermentation of rhHSP70

[0082]To prepare sufficient amounts of HSP70, 5 L of the E. coli expression strain HMS 174 (DE3) hHSP70 may be grown in animal free medium containing 12 g phytone peptone, 60 g yeast extract, 40 g sodium chloride, 1g methionine, and water to 5 liters. Before inoculation with expression strain bacteria, the medium may be sterilized, and supplemented with kanamycin to a final concentration of 30 mcg per milliliter. The medium may then be inoculated with one liter of an overnight culture of the HMS174(DE3) bacteria. The pH, level of dissolved oxygen and temperature inside the fermenter reactor may be carefully monitored. When the culture reaches an optical density of approximately 1.2 (for example, approximately 2.5 hours after inoculation), IPTG (isopropyl-B-D-thiogalactopyranoside) may be added to a final concentration of 1 mM to initiate the induction of expression of rhHSP70. Growth may then be continued until an optical density of approximately 2.7 is attain...

example 2

Purification of rhHSP70

[0083]A bacterial lysate comprising rhHSP70 may be subjected to the following two-step purification protocol. The specific example provided utilizes 500 mL of lysate; adjustments may be made for different lysate volumes. In the first step of the protocol, 500 mL of the cleared lysate, prepared as set forth above, may be diluted two-fold with 10 mM sodium phosphate buffer pH 7.0. This 1 L of diluted and cleared bacterial lysate may be loaded onto a of DEAE Sephacel (Pharmacia) column (13 cmx 15 cm) previously equilibrated with 2 column volumes of buffer A (10 mM sodium phosphate buffer pH 7.0, 20 mM sodium chloride, 10 mM ammonium sulfate). The elution is carried out with two column volumes of buffer B (20 mM sodium phosphate buffer pH 7, 85 mM sodium chloride, 10 mM ammonium sulfate). About 2 L of eluate is expected for recovery. The eluate may then be diafiltered (buffer exchanged) against a 10-fold volume of buffer C (20 mM sodium citrate pH 6.0, 100 mM sodi...

example 3

PEGylation Techniques

[0085]The efficacy of a therapeutic agent may be enhanced by improving its bioavailability via several means, one of which is PEGylation, a process of chemically linking polyethylene glycol (PEG) to the therapeutic agent of interest, with the resulting conjugate exhibiting an increased serum half-life. Additional advantages of the PEGylated products may also include lower immunogenicity, decreased dosing frequency, increased solubility, enhanced stability, and reduced renal clearance. Because the most common reactive sites on proteins (including peptides) for attaching PEG are the c amino groups of lysine and the a amino group of the N-terminal residue, early methods of PEGylation resulted in modification of multiple sites, yielding not only monoPEGylated conjugates consisting of mixtures of positional isomers, such as PEGINTRONTM (Grace et al., J. Biol. Chem. 2005; 280:6327) and PEGASYS® (Dhalluin et al., Bioconjugate Chem. 2005; 16:504), but also adducts compr...

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Abstract

The present invention relates to novel therapies that utilize exogenous HSP70 for the treatment of disorders or conditions regulated by HSP70 through administration of exogenous HSP70.

Description

RELATED APPLICATIONS[0001]This application is a US Utility Application that claims the benefit of priority from U.S. Provisional Patent Application No. 61 / 502,332, filed on Jun. 28, 2011, the entirety of which is incorporated herein by reference.BACKGROUND OF THE INVENTION[0002]Over 6 million Americans are diagnosed annually with musculoskeletal disease, which is a major cause of work disability. Annual costs associated with treatment and care of musculoskeletal disease is in the tens of billions of dollars. Muscle degeneration and impairment in such disease may be stemming from various causes, including tissue ischemia, severe injury, advanced age, and muscular dystrophy resulting form genetic defects as well as major diseases such as cancer or kidney failure.[0003]Current key therapeutic approaches and standards of care are typically limited to physiotherapeutic rehabilitation, pain management and anti-inflammatory medications. Lately, there has been an increased interest in devel...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61K38/17
CPCA61K38/1703A61K38/17A61K47/48215A61K47/60A61P21/00
Inventor ONIKIENKO, SERGEI B.NIVOROZHKIN, ALEXZEMLYANOI, ALEXANDER V.
Owner ALTERNATIVE INNOVATIVE TECH
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