Granted Patent B2

US 9,910,047 · App. 14/763,962 · Granted Mar 6, 2018

Abstract

Claims (41)

1. A method for generating a risk score useful for determining the probability that a subject will progress from being normal to having one or more disorders selected from the group consisting of Insulin Resistance, Impaired Glucose Tolerance, Type-2 diabetes, and CVD, the method comprising:

producing an analytical sample by extracting small molecules from a biological sample from the subject;

determining the levels of cinnamoylglycine, 5alpha-androstan-3beta,17alpha-diol disulfate, adrenate and 3-hydroxypropanoate; and

analyzing the levels of cinnamoylglycine, adrenate, 5alpha-androstan-3beta,17alpha-diol disulfate and 3-hydroxypropanoate in the sample by a statistical analysis to generate a risk score, wherein the risk score can be used to aid in determining the probability that the subject will progress to one or more disorders selected from Insulin Resistance, CVD, Type 2 diabetes, and Impaired Glucose Tolerance.

2. The method of claim 1 , wherein the statistical analysis comprises a logistic regression model.

3. The method of claim 1 , wherein the risk score is generated to aid in predicting the progression to cardiovascular disease.

4. The method of claim 1 , further comprising:

determining the level(s) of one or more additional small molecule biomarkers in the biological sample, wherein the additional biomarkers are selected from the group consisting of 7-alpha-hydroxy-3-oxo-4-cholestenoate (7-Hoca), 3 hydroxy-2-oxovalerate, beta-hydroxypyruvate, palmitoyl sphingomyelin, oleoylcarnitine, 1-methylurate, 1,3-dimethylurate, 1,7-dimethylurate, 1,3,7-trimethylurate, and 5alpha-pregnan-3beta, 20alpha-diol disulfate; and

analyzing the levels of cinnamoylglycine, adrenate, 5alpha-androstan-3beta,17alpha-diol disulfate and 3-hydroxypropanoate and the level(s) of the one or more additional biomarkers in the sample by a statistical analysis to generate a risk score, wherein the risk score can be used to aid in determining the probability that the subject will progress to one or more disorders selected from Insulin Resistance, CVD, Impaired Glucose Tolerance and Type 2 diabetes.

5. The method of claim 4 , wherein the analyzing step comprises generating a probability curve using a multivariate regression model based on the measured levels of cinnamoylglycine, adrenate, 5alpha-androstan-3beta,17alpha-diol disulfate and 3-hydroxypropanoate; and level(s) of the one or more additional biomarkers.

6. The method of claim 5 , wherein the probability curve is generated using a multivariate regression model based on the measured levels of cinnamoylglycine, adrenate, 7-alpha-hydroxy-3-oxo-4-cholestenoate (7-Hoca), 3-hydroxypropanoate, 3-hydroxy-2-oxovalerate, beta-hydroxypyruvate, palmitoyl sphingomyelin, oleoylcarnitine, 1-methylurate, 1,3-dimethylurate, 1,7-dimethylurate, 1,3,7-trimethylurate, 5 alpha-androstan-3beta,17alpha-diol disulfate, and 5alpha-pregnan-3beta, 20alpha-diol disulfate.

7. The method of claim 5 , wherein the probability curve is generated using a multivariate regression model using the measured levels of the cinnamoylglycine, adrenate, 5alpha-androstan-3beta,17alpha-diol disulfate and 3-hydroxypropanoate and one or more additional biomarkers selected from the group consisting of 7-alpha-hydroxy-3-oxo-4-cholestenoate (7-Hoca), 3 hydroxy-2-oxovalerate, beta-hydroxypyruvate, palmitoyl sphingomyelin, oleoylcarnitine, 1-methylurate, 1,3-dimethylurate, 1,7-dimethylurate, 1,3,7-trimethylurate, and 5alpha-pregnan-3beta,20alpha-diol disulfate.

8. The method of claim 4 , wherein the risk score is generated to aid in predicting the progression to Type-2 diabetes and the one or more additional biomarker comprises 7-alpha-hydroxy-3-oxo-4-cholestenoate (7-Hoca).

9. A method of generating a risk score to aid in determining susceptibility of a subject to type-2 diabetes, the method comprising:

producing an analytical sample by extracting small molecules from a biological sample from the subject;

analyzing the analytical sample from the subject to determine the levels of cinnamoylglycine, adrenate, 5alpha-androstan-3beta,17alpha-diol disulfate and 3-hydroxypropanoate; and

analyzing the levels of cinnamoylglycine, adrenate, 5alpha-androstan-3beta,17alpha-diol disulfate and 3-hydroxypropanoate by a statistical analysis to generate a risk score, wherein the risk score aids in determining whether the subject is susceptible to developing type-2 diabetes.

10. The method of claim 9 , further comprising:

determining the level(s) of one or more additional small molecule biomarkers in the biological sample, wherein the additional biomarkers are selected from the group consisting of 7-alpha-hydroxy-3-oxo-4-cholestenoate (7-Hoca), 3 hydroxy-2-oxovalerate, beta-hydroxypyruvate, palmitoyl sphingomyelin, oleoylcarnitine, 1-methylurate, 1,3-dimethylurate, 1,7-dimethylurate, 1,3,7-trimethylurate, and 5alpha-pregnan-3beta, 20alpha-diol disulfate; and

analyzing the levels of cinnamoylglycine, adrenate, 5alpha-androstan-3beta,17alpha-diol disulfate and 3-hydroxypropanoate and the level(s) of the one or more additional biomarkers in the sample by a statistical analysis to generate a risk score, wherein the risk score aids in determining whether the subject is susceptible to developing type-2 diabetes.

11. The method of claim 10 , wherein the one or more additional biomarkers comprise 7-alpha-hydroxy-3-oxo-4-cholestenoate (7-Hoca).

12. A method of determining susceptibility of a subject to cardiovascular disease, the method comprising:

analyzing the biological sample from the subject to determine the levels of cinnamoylglycine, adrenate, 5 alpha-androstan-3beta,17alpha-diol di sulfate and 3-hydroxypropanoate; and

comparing the levels of cinnamoylglycine, adrenate, 5alpha-androstan-3beta,17alpha-diol disulfate and 3-hydroxypropanoate to CVD-positive and/or CVD-negative reference levels thereof in order to determine whether the subject is susceptible to developing cardiovascular disease.

13. The method of claim 12 , further comprising:

analyzing the biological sample from the subject to determine the levels of cinnamoylglycine, adrenate, 5alpha-androstan-3beta,17alpha-diol disulfate and 3-hydroxypropanoate and one or more additional biomarkers selected from the group consisting of 7-alpha-hydroxy-3-oxo-4-cholestenoate (7-Hoca), 3 hydroxy-2-oxovalerate, beta-hydroxypyruvate, palmitoyl sphingomyelin, oleoylcarnitine, 1-methylurate, 1,3-dimethylurate, 1,7-dimethylurate, 1,3,7-trimethylurate, and 5alpha-pregnan-3beta, 20alpha-diol disulfate; and

comparing the levels of cinnamoylglycine, adrenate, 5alpha-androstan-3beta,17alpha-diol disulfate and 3-hydroxypropanoate and the one or more additional biomarkers in the sample to CVD-positive and/or CVD-negative reference levels thereof in order to determine whether the subject is susceptible to developing cardiovascular disease.

14. The method of claim 13 , wherein the cardiovascular disease is selected from the group consisting of myocardial infarction and stroke.

15. The method of claim 13 , wherein the comparing step comprises generating a cardiovascular disease risk score for the subject in order to determine the susceptibility to cardiovascular disease in the subject.

16. A method of generating a risk score to aid in monitoring the progression or regression of insulin resistance in a subject, the method comprising:

producing an analytical sample by extracting small molecules from a biological sample from the subject;

analyzing the analytical sample from the subject to determine the levels of cinnamoylglycine, adrenate, 5alpha-androstan-3beta,17alpha-diol di sulfate and 3-hydroxypropanoate; and

analyzing the levels of cinnamoylglycine, adrenate, 5alpha-androstan-3beta,17alpha-diol disulfate and 3-hydroxypropanoate in the sample by a statistical analysis to generate a risk score, wherein the risk score is used to aid in monitoring the progression or regression of insulin resistance in the subject.

17. The method of claim 16 , further comprising:

analyzing the biological sample from the subject to determine the levels of cinnamoylglycine, adrenate, 5alpha-androstan-3beta,17alpha-diol disulfate and 3-hydroxypropanoate and one or more additional small molecule biomarkers selected from the group consisting of 7-alpha-hydroxy-3-oxo-4-cholestenoate (7-Hoca), 3 hydroxy-2-oxovalerate, beta-hydroxypyruvate, palmitoyl sphingomyelin, oleoylcarnitine, 1-methylurate, 1,3-dimethylurate, 1,7-dimethylurate, 1,3,7-trimethylurate, and 5alpha-pregnan-3beta,20alpha-diol disulfate; and

analyzing the levels of cinnamoylglycine, adrenate, 5alpha-androstan-3beta,17alpha-diol disulfate and 3-hydroxypropanoate and the one or more additional biomarkers in the sample by a statistical analysis to generate a risk score, wherein the risk score is used to aid in monitoring the progression or regression of insulin resistance in the subject.

18. The method of claim 17 , wherein the subject is selected from the group consisting of a subject being treated with a pharmaceutical composition, a subject having undergone bariatric surgery, a subject undergoing an exercise modification, and a subject using a dietary modification.

19. The method of claim 17 , wherein the analyzing step comprises transforming the biological sample by protein extraction.

20. The method of claim 17 , wherein the disease risk score is a cardiovascular disease risk score.

21. The method of claim 17 , wherein the disease risk score is a type 2 diabetes risk score.

22. The method of claim 21 wherein the one or more additional biomarkers comprise 7-alpha-hydroxy-3-oxo-4-cholestenoate (7-Hoca).

Assignments (10)