ADJUSTING EXPRESSION LEVEL OF A GENE ENCODING A SIRTUIN PROTEIN BY TREATING A HUMAN SUBJECT WITH A NITROXIDE

Abstract

Some embodiments disclosed herein include a method for increasing an expression level of a gene. The methods include administering an effective amount of a nitroxide antioxidant to a human subject having a decreased expression level of SIRT2, whereby expression level of the gene is increased.

Claims

1. A method of increasing expression level of a gene encoding SIRT2, the method comprising: identifying a subject having a disease or condition associated with decreased expression level of the gene encoding SIRT2, the disease or condition selected from cardiac hypertrophy, a neurological disorder, and atherosclerosis; and administering an effective amount of a nitroxide antioxidant to the subject, whereby the expression level of the gene encoding SIRT2 is increased.

2. (canceled)

3. The method of claim 1, wherein upregulation of SIRT2 reverses an age-related inhibition of SIRT2 activity, wherein the age-related inhibition of SIRT2 activity is associated with the decreased expression of SIRT2.

4. (canceled)

5. The method of claim 1, wherein the disease or condition is cardiac hypertrophy.

6. (canceled)

7. The method of claim 1, disease or condition is a neurological disorder associated with increased Ehmt2 expression, and whereby the increased expression of SIRT2 reduces expression of Ehmt2.

8. The method of claim 1, wherein the disease or condition is atherosclerosis.

9. (canceled)

10. A method for increasing expression level of a gene encoding SIRT2, the method comprising: identifying a subject having a disease or condition associated with decreased expression of SIRT2, the disease or condition selected from cardiac hypertrophy, a neurological disorder, and atherosclerosis; and administering an effective amount of a nitroxide antioxidant to the subject, wherein the expression level of the gene encoding SIRT2 is increased.

11. The method of claim 10, wherein the nitroxide antioxidant is 4-hydroxy-2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPOL).

12. (canceled)

13. (canceled)

14. (canceled)

15. The method of claim 10, whereby the increased expression level of SIRT2 activates NRF2.

16. A method for increasing expression level of SIRT2, the method comprising: administering a nitroxide antioxidant to a subject having a disease or condition associated with a decreased expression level of SIRT2, the disease or condition selected from cardiac hypertrophy, a neurological disorder, and atherosclerosis, whereby the expression level of the gene is increased.

17. (canceled)

18. A method for increasing the expression level of SIRT2 in a human subject in need thereof, comprising: administering to a human subject, known to have [[an]]a decreased expression of SIRT2 gene associated with a disease, an effective amount of a nitroxide antioxidant to increase the level of expression of the SIRT2 gene associated with the disease, wherein the disease or condition is selected from cardiac hypertrophy, a neurological disorder, and atherosclerosis, wherein the expression level of the gene encoding SIRT2 is increased in cardiac tissue.

19. The method of claim 18, wherein the nitroxide antioxidant is 4-hydroxy-2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPOL).

20. The method of claim 18, wherein the expression level of the gene encoding SIRT2 is increased in cardiac tissue.

21. The method of claim 1, wherein the nitroxide antioxidant is selected from the group consisting of 2-ethyl-2,5,5-trimethyl-3-oxazolidine-1-oxyl (OXANO), 2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPO), 4-hydroxy-2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPOL), 4-amino-2,2,6,6-tetramethyl-1-piperidinyloxy (Tempamine), 3-Amin omethyl-PROXYL, 3-Cyano-PROXYL, 3-Carbamoyl-PROXYL, 3-Carboxy-PROXYL, and 4-Oxo-TEMPO. TEMPO can also be substituted, typically in the 4 position, for example, 4-amino, 4-(2-bromoacetamido), 4-(ethoxyfluorophosphonyloxy), 4-hydroxy, 4-(2-iodoacetamido), 4-isothiocyanato, 4-maleimido, 4-(4-nitrobenzoyloxyl), 4-phosphonooxy, 2,2,6,6-tetramethyl-4-oxo-1-piperidinyloxy (TEMPONE), 1-Hydroxy-2,2,6,6-tetramethyl-4-oxo-piperidine HCl (TEMPONE-H), 1,2-dipalmitoyl-sn-glycero-3-phospho(tempo)choline (TEMPO PC), (4-[N,N-dimethyl-N-(2-hydroxyethyl)]ammonium-2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPO Choline).

22. The method of claim 1, wherein the expression level of the gene encoding SIRT2 is increased in cardiac tissue.

23. The method of claim 1, wherein the nitroxide antioxidant is 4-hydroxy-2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPOL).

24. The method of claim 16, wherein the nitroxide antioxidant is 4-hydroxy-2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPOL).

25. The method of claim 16, wherein the expression level of the gene encoding SIRT2 is increased in cardiac tissue.

26. The method of claim 10, wherein the expression level of the gene encoding SIRT2 is increased in cardiac tissue.

Description

EXAMPLES

[0123] Some aspects of the embodiments discussed above are disclosed in further detail in the following examples, which are not in any way intended to limit the scope of the present disclosure.

[0124] In order to facilitate understanding, the specific embodiments are provided to help interpret the technical proposal, that is, these embodiments are only for illustrative purposes, but not in any way to limit the scope of the invention. Unless otherwise specified, embodiments do not indicate the specific conditions, are in accordance with the conventional conditions or the manufacturer's recommended conditions.

Example 1. Effects of Tempol on Expression of Genes Associated With Sirtuin Protein Activity

[0125] To assess the effects of Tempol on gene expression, Tempol was administered to experimental mice at a dose of 5 mg/g of food from 14 months to 31 months after birth. Mice receiving the same food without the addition of Tempol were used as a negative control. At the age of 31 months, the experimental animals were sacrificed and the hearts were surgically removed. The expression of a broad spectrum of genes in the cardiac tissue was assessed using chip-based microarray technology. Such chips are well known in the art and are widely used to assess gene expression. The experimental results showed that SIRT2 exhibited statistically significant increase in expression. This result is shown in Table 1.

TABLE-US-00001 TABLE 1 Genes Associated With SIRT2 Exhibiting Increased Expression In Cardiac Tissue After Tempol Administration Symbol Gene title Fold change P-value SIRT2 Sirtuin 2 1.16 0.00

Example 2. Treating Age-Related Increase in Gene Expression

[0126] A 70-kilogram human subject over the age of 65 is identified as having, or known to have, or suspected of having a decreased expression level of SIRT2. The human subject is administered a dose of 2000 mg of Tempol (or another nitroxide antioxidant) per day for 180 days. This may be administered in a single dose, or may be administered as a number of smaller doses over a 24-hour period: for example, four 500-mg doses at eight-hour intervals. Following treatment, the serum level of SIRT2, is increased.

Example 3. Treating a Human Subject with Increased Gene Expression

[0127] A 70-kilogram human subject is identified as having, or known to have, or suspected of having a decreased expression level of SIRT2. The human subject is administered a dose of 2000 mg of Tempol (or another nitroxide antioxidant) per day for 180 days. This may be administered in a single dose, or may be administered as a number of smaller doses over a 24-hour period: for example, four 500-mg doses at eight-hour intervals. Following treatment, the serum level of SIRT2, is increased.

Example 4. Treating a Human Subject with an Age-Related Disease

[0128] A 70-kilogram human subject over the age of 65 and having a cardiovascular disease is identified for a decreased expression level of SIRT2. Or a 70-kilogram human subject over the age of 65 is known to have a cardiovascular disease and/or decreased expression level of SIRT2. The human subject is administered a dose of 2000 mg of Tempol (or another nitroxide antioxidant) per day for 180 days. This may be administered in a single dose, or may be administered as a number of smaller doses over a 24-hour period: for example, four 500-mg doses at eight-hour intervals. Following treatment, the serum level of SIRT2, is increased.

Example 5. Treating a Human Subject at Risk of Developing Cancer

[0129] A 70-kilogram human subject at risk of developing colorectal cancer is identified for decreased expression level of SIRT2. Or a 70-kilogram human subject is known to be at risk of developing colorectal cancer and/or have decreased expression level of SIRT2. The human subject is administered a dose of 2000 mg of Tempol (or another nitroxide antioxidant) per day for 180 days. This may be administered in a single dose, or may be administered as a number of smaller doses over a 24-hour period: for example, four 500-mg doses at eight-hour intervals. Following treatment, the serum level of SIRT2, is increased.

Example 6. Treating a Human Subject at Risk of Developing an Autoimmune Disease

[0130] A 70-kilogram human subject at risk of developing an autoimmune disease (e.g., rheumatoid arthritis) is identified for decreased expression level of SIRT2. Or a 70-kilogram human subject is known to be at risk of developing an autoimmune disease and/or have decreased expression level of SIRT2. The human subject is administered a dose of 2000 mg of Tempol (or another nitroxide antioxidant) per day for 180 days. This may be administered in a single dose, or may be administered as a number of smaller doses over a 24-hour period: for example, four 500-mg doses at eight-hour intervals. Following treatment, the serum level of SIRT2, is increased.

Example 7. Treating a Human Subject at Risk of Developing a Condition Due to Aging

[0131] A 70-kilogram human subject of 45 years old at risk of developing a condition due to aging is identified. Or a 70-kilogram human subject of 45 years old is known to be at risk of developing a condition. The human subject is administered a dose of 2000 mg of Tempol (or another nitroxide antioxidant) per day for 180 days. This may be administered in a single dose, or may be administered as a number of smaller doses over a 24-hour period: for example, four 500-mg doses at eight-hour intervals. Following treatment, the serum level of SIRT2, is increased.

Example 8. Treating a Human Subject at Risk of Developing a Neruodegenerative Disease

[0132] A 70-kilogram human subject at risk of developing a neurodegenerative disease (e.g., Parkinson's Disease) is identified for decreased expression level of SIRT2. Or a 70-kilogram human subject is known to be at risk of developing a neurodegenerative disease and/or have decreased expression level of SIRT2. The human subject is administered a dose of 2000 mg of Tempol (or another nitroxide antioxidant) per day for 180 days. This may be administered in a single dose, or may be administered as a number of smaller doses over a 24-hour period: for example, four 500-mg doses at eight-hour intervals. Following treatment, the serum level of SIRT2, is increased.

Example 9. Treating a Human Subject Having an Infection

[0133] A 70-kilogram human subject having an infection (e.g., a bacterial, fungal, or viral infection) is identified for decreased expression level of SIRT2. Or a 70-kilogram human subject is known to have an infection and/or have decreased expression level of SIRT2. The human subject is administered a dose of 2000 mg of Tempol (or another nitroxide antioxidant) per day for 180 days. This may be administered in a single dose, or may be administered as a number of smaller doses over a 24-hour period: for example, four 500-mg doses at eight-hour intervals. Following treatment, the serum level of SIRT2, is increased.

[0134] In at least some of the previously described embodiments, one or more elements used in an embodiment can interchangeably be used in another embodiment unless such a replacement is not technically feasible. It will be appreciated by those skilled in the art that various other omissions, additions and modifications may be made to the methods and structures described above without departing from the scope of the claimed subject matter. All such modifications and changes are intended to fall within the scope of the subject matter, as defined by the appended claims.

[0135] With respect to the use of substantially any plural and/or singular terms herein, those having skill in the art can translate from the plural to the singular and/or from the singular to the plural as is appropriate to the context and/or application. The various singular/plural permutations may be expressly set forth herein for sake of clarity.

[0136] It will be understood by those within the art that, in general, terms used herein, and especially in the appended claims (e.g., bodies of the appended claims) are generally intended as “open” terms (e.g., the term “including” should be interpreted as “including but not limited to,” the term “having” should be interpreted as “having at least,” the term “includes” should be interpreted as “includes but is not limited to,” etc.). It will be further understood by those within the art that if a specific number of an introduced claim recitation is intended, such an intent will be explicitly recited in the claim, and in the absence of such recitation no such intent is present. For example, as an aid to understanding, the following appended claims may contain usage of the introductory phrases “at least one” and “one or more” to introduce claim recitations. However, the use of such phrases should not be construed to imply that the introduction of a claim recitation by the indefinite articles “a” or “an” limits any particular claim containing such introduced claim recitation to embodiments containing only one such recitation, even when the same claim includes the introductory phrases “one or more” or “at least one” and indefinite articles such as “a” or “an” (e.g., “a” and/or “an” should be interpreted to mean “at least one” or “one or more”); the same holds true for the use of definite articles used to introduce claim recitations. In addition, even if a specific number of an introduced claim recitation is explicitly recited, those skilled in the art will recognize that such recitation should be interpreted to mean at least the recited number (e.g., the bare recitation of “two recitations,” without other modifiers, means at least two recitations, or two or more recitations). Furthermore, in those instances where a convention analogous to “at least one of A, B, and C, etc.” is used, in general such a construction is intended in the sense one having skill in the art would understand the convention (e.g., “a system having at least one of A, B, and C” would include but not be limited to systems that have A alone, B alone, C alone, A and B together, A and C together, B and C together, and/or A, B, and C together, etc.). In those instances where a convention analogous to “at least one of A, B, or C, etc.” is used, in general such a construction is intended in the sense one having skill in the art would understand the convention (e.g., “a system having at least one of A, B, or C” would include but not be limited to systems that have A alone, B alone, C alone, A and B together, A and C together, B and C together, and/or A, B, and C together, etc.). It will be further understood by those within the art that virtually any disjunctive word and/or phrase presenting two or more alternative terms, whether in the description, claims, or drawings, should be understood to contemplate the possibilities of including one of the terms, either of the terms, or both terms. For example, the phrase “A or B” will be understood to include the possibilities of “A” or “B” or “A and B.”

[0137] In addition, where features or aspects of the disclosure are described in terms of Markush groups, those skilled in the art will recognize that the disclosure is also thereby described in terms of any individual member or subgroup of members of the Markush group.

[0138] As will be understood by one skilled in the art, for any and all purposes, such as in terms of providing a written description, all ranges disclosed herein also encompass any and all possible sub-ranges and combinations of sub-ranges thereof. Any listed range can be easily recognized as sufficiently describing and enabling the same range being broken down into at least equal halves, thirds, quarters, fifths, tenths, etc. As a non-limiting example, each range discussed herein can be readily broken down into a lower third, middle third and upper third, etc. As will also be understood by one skilled in the art all language such as “up to,” “at least,” “greater than,” “less than,” and the like include the number recited and refer to ranges which can be subsequently broken down into sub-ranges as discussed above. Finally, as will be understood by one skilled in the art, a range includes each individual member. Thus, for example, a group having 1-3 articles refers to groups having 1, 2, or 3 articles. Similarly, a group having 1-5 articles refers to groups having 1, 2, 3, 4, or 5 articles, and so forth.

[0139] While various aspects and embodiments have been disclosed herein, other aspects and embodiments will be apparent to those skilled in the art. The various aspects and embodiments disclosed herein are for purposes of illustration and are not intended to be limiting, with the true scope and spirit being indicated by the following claims.