초록 ▼

A method to assess arachidonic acid (AA) metabolites-dependent hypertension by measuring glucuronidated dihydroxyeicosatrienoic acids (DHETs) and DHET metabolites in a biological sample which contains the epitopes unique to DHET (using any methods including GC/MS, LC/MS or ELISA) is disclosed. An ex

A method to assess arachidonic acid (AA) metabolites-dependent hypertension by measuring glucuronidated dihydroxyeicosatrienoic acids (DHETs) and DHET metabolites in a biological sample which contains the epitopes unique to DHET (using any methods including GC/MS, LC/MS or ELISA) is disclosed. An example of the glucuronidated DHET metabolite is DHET-alcohols such as omega or omega-1 oxidated DHET and DHET esterified glycerol. The method further includes determining the amount of glucuronidated molecules containing a DHET-specific epitope which is immunoreactive with antibodies produced against DHETs. The present invention measuring glucuronidated DHET levels in a biological sample is useful for drug development and monitoring efficiency of drug treatment of a mammal who has AA epoxygenase-, epoxide hydrolase-and/or UDP-glucuronosyl transferase-dependent hypertension.

대표청구항 ▼

The invention claimed is: 1. A method for assessing the presence of hypertension in a mammal comprising the steps of measuring an amount of glucuronidated dihydroxyeicosatrienoic acids (DHETs) from a test sample from a mammal; comparing the amount of glucuronidated dihydroxyeicosatrienoic acids in

The invention claimed is: 1. A method for assessing the presence of hypertension in a mammal comprising the steps of measuring an amount of glucuronidated dihydroxyeicosatrienoic acids (DHETs) from a test sample from a mammal; comparing the amount of glucuronidated dihydroxyeicosatrienoic acids in the test sample to a control sample from a non-hypertension mammal of a same species and determining if the test sample contains a comparatively elevated amount of the glucuronidated DHETs as an indication of hypertension mediated by one or both of epoxide hydrolase and UDP-glucuronosyl transferase; wherein the test sample is selected from the group consisting of urine, blood, liver and kidney. 2. The method as defined in claim 1, wherein said measuring an amount of glucuronidated dihydroxyeicosatrienoic acids includes the steps of: obtaining a test specimen from said test sample; adding glucuronidase to the test specimen; collecting a first sample from the test specimen at time zero of glucuronidase digestion, and a second sample after digestion is completed immunoassaying the first and second samples with antibody immunoreactive with a specific DHET; and determining the net amount of the specific DHET that is glurcuronidated by subtracting the amount of immunoreactive DHET in the first sample from the amount of immunoreactive DHET in the second sample. 3. The method as defined in claim 2, wherein said antibodies are immunoreactive with 14,15-DHETs. 4. The method as defined in claim 1, further including the step of identifying patients with hypertension. 5. The method as defined in claim 4, wherein said patient is a pregnant woman. 6. The method as defined in claim 5, wherein said patient has pregnancy-induced hypertension. 7. The method as defined in claim 1, wherein said test sample is in a urine sample. 8. The method as defined in claim 1, wherein said measuring step is further defined as measuring glucuronidated DHET metabolites. 9. The method as defined in claim 8, wherein said measuring of glucuronidated DHET metabolites includes the steps of: obtaining a test specimen from said test sample; adding glucuronidase to the test specimen; collecting a first sample from the test specimen at time zero of glucuronidase digestion, and a second sample after digestion is completed immunoassaying the first and second samples with antibody immunoreactive with a specific DHET metabolite; and determining the net amount of the specific DHET metabolite that is glurcuronidated by subtracting the amount of immunoreactive DHET metabolite in the first sample from the amount of immunoreactive DHET metabolite in the second sample. 10. The method as defined in claim 9, wherein said antibodies are immunoreactive with metabolites of 14,15-DHET. 11. A method for determining DHET-dependent UDP-glycuronosyl transferase activity in a biological system including the steps of: converting a known amount of a DHET to the corresponding glucuronidated DHET by UDP-glycuronosyl transferase activity in a biological system; collecting a first sample and a second sample; adding glucuronidase to the first sample and allowing complete digestion; leaving untreated the second sample; immunoassaying the first sample and the second sample with antibody immunoreactive with a specific DHET; determining the net amount of the specific DHET that is glurcuronidated by subtracting the amount of immunoreactive DHET in the first sample from the amount of immunoreactive DHET in the second sample; and determining DHET-dependent UDP-glururonosyl transferase activity. 12. The method as defined in claim 11, wherein said antibody immunoreactive with a specific DHET is immunoreactive with a DHET selected from the group comprising 5,6-DHET, 8,9-DHET, 14,15-DHET, and 11,12-DHET. 13. The method as defined in claim 1, wherein said measuring an amount of glucuronidated dihydroxyeicosatrienoic acids includes the steps of: obtaining a test specimen from said test sample; immunoassaying a first sample of the test specimen with an antibody immunoreactive with the glucuronidated form of a specific DHET; immunoassaying a second sample of the test specimen with an antibody immunoreactive with the free form of the specific DHET; and determining the total (free+glucuronidated) amount of the specific DHET by adding the amount of immunoreactive glucuronidated DHET in the first sample to the amount of immunoreactive free DHET in the second sample. 14. The method as defined in claim 13 wherein the antibody immunoreactive with the glucuronidated form of a specific DHET is an antibody immunoreactive with the glucuronidated form of a DHET selected from the group comprising 5,6-DHET, 8,9-DHET, 14,15-DHET, and 11,12-DHET, and the antibody immunoreactive with the free form of a specific DHET is an antibody immunoreactive with the free form of a DHET selected from the group comprising 5,6-DHET, 8,9-DHET, 14,15-DHET, and 11,12-DHET. 15. The method as defined in claim 8, wherein said measuring of glucuronidated DHET metabolites includes: obtaining a test specimen from said test sample; immunoassaying a first sample of the test specimen with an antibody immunoreactive with the glucuronidated form of a specific DHET metabolite; immunoassaying a second sample of the test specimen with an antibody immunoreactive with the free form of the specific DHET metabolite; and determining the total (free+glucuronidated) amount of the specific DHET metabolite by adding the amount of immunoreactive glucuronidated DHET metabolite in the first sample to the amount of immunoreactive free DHET metabolite in the second sample. 16. The method as defined in claim 15 wherein the antibody immunoreactive with the glucuronidated form of a specific DHET metabolite is an antibody immunoreactive with the glucuronidated form of a DHET metabolite selected from the group comprising 5,6-DHET, 8,9-DHET, 14,15-DHET, and 11,12-DHET, and the antibody immunoreactive with the free form of a specific DHET metabolite is an antibody immunoreactive with the free form of a DHET metabolite selected from the group comprising 5,6-DHET, 8,9-DHET, 14,15-DHET, and 11,12-DHET.