Compositions and methods to protect mammalian tissue against cold and other metabolic stresses

Patent metadata

Field	Value
Publication number	US-11547708-B2
Application number	US-201816636945-A
Country	US
Kind code	B2
Filing date	Aug 20, 2018
Priority date	Aug 21, 2017
Publication date	Jan 10, 2023
Grant date	Jan 10, 2023

Abstract

A composition for preserving viability of cells, tissues, or organs at a low temperature is provided. The composition includes a mitochondrial uncoupling agent, at least one protease inhibitor, and a reducing agent. Methods to protect cells, tissues, or organs from exposure to cold and other metabolic stress are also provided.

First claim

What is claimed is:

1. A method for treating mammalian cells, tissues, or organs in need of protection from low-temperature injury, the method comprising:
contacting the mammalian cells, tissues, or organs with a composition comprising:
a compound of formula (I)

a protease inhibitor selected from the group consisting of a cysteine protease inhibitor, an aspartic protease inhibitor, a serine protease inhibitor, and a leucine aminopeptidase inhibitor and
a reducing agent; and

exposing the mammalian cells, tissues, or organs to a temperature of from about 1-5° C.

2. The method according to claim 1 , wherein the reducing agent is ascorbic acid.

3. The method according to claim 1 , wherein the protease inhibitor is a combination of E-64, Pepstatin A, AEBSF, and Bestatin, and the reducing agent is ascorbic acid.

4. The method according to claim 1 , further comprising admixing the composition with a cryopreservation agent to form a solution, wherein contacting the mammalian cells, tissues, or organs with the composition comprises contacting the mammalian cells, tissues, or organs with the solution.

5. The method according to claim 4 , wherein the composition comprises the compound of Formula (I), E-64, Pepstatin A, AEBSF, Bestatin, and ascorbic acid, the solution having
a concentration of the compound of Formula (I) from 0.05 to 0.2 μM;
a concentration of E-64 from 3 to 5 μM;
a concentration of Pepstatin A from 2 to 4 μM;
a concentration of AEBSF from 100 to 200 μM;
a concentration of Bestatin from 10 to 20 μM; and
a concentration of ascorbic acid from 50 to 300 μM.

6. The method according to claim 1 , wherein the mammalian cells, tissues, or organs are selected from the group consisting of neuronal cells, retinal cells, corneal cells, corneal tissue, a cornea, retina tissue, a retina, skin cells, skin tissue, skin, heart cells, heart tissue, a heart, lung cells, lung tissue, a lung, pancreal cells, pancreas tissue, a pancreas, kidney cells, kidney tissue, a kidney, liver cells, liver tissue, a liver, intestinal cells, intestinal tissue, intestine, stem cells, blood cells and blood.

7. The method according to claim 1 , wherein the temperature is from 2° C. to 5° C.

8. The method according to claim 7 wherein the treatment has a duration from 4 to 72 hours.

9. The method according to claim 4 wherein the cryopreservation agent is a solution comprising hydroxyethyl starch 50.0 g/L; lactobionic acid (as Lactone) 35.83 g/L; potassium dihydrogen phosphate 3.4 g/L; magnesium sulfate heptahydrate 1.23 g/L; raffinose pentahydrate 17.83 g/L; adenosine 1.34 g/L; allopurinol 0.136 g/L; total glutathione 0.922 g/L; and potassium hydroxide 5.61 g/L, adjusted to pH 7.4.

10. The method according to claim 1 wherein the protease inhibitor consists essentially of the peptide Iva-Val-Val-Sta-Ala-Sta (Pepstatin A).

11. The method accordingly claim 9 wherein the hydroxyethyl starch comprises Pentafraction.

Assignees

• Us Health
• The Usa As Represented By The Secretary Dhhs

Inventors

• Ou Jingxing
• Li Wei
• Miyagishima Kiyoharu Josh

Classifications

• A61K31/5395

  having two or more nitrogen atoms in the same ring, e.g. oxadiazines · CPC title

• A61K45/06 Primary

  Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca · CPC title

• A61K31/197

  the amino and the carboxyl groups being attached to the same acyclic carbon chain, e.g. gamma-aminobutyric acid [GABA], beta-alanine, epsilon-aminocaproic acid or pantothenic acid (carnitine A61K31/205) · CPC title

• A61K38/55

  Protease inhibitors · CPC title

• A61K31/34

  having five-membered rings with one oxygen as the only ring hetero atom, e.g. isosorbide · CPC title