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Methods of determining active levels of drugs in fluid samples

a technology of active levels and fluid samples, applied in the field of screening for the presence of drugs in fluid samples, can solve the problems of not providing the needed information, and achieve the effects of regulating blood pressure, regulating blood pressure, and regulating renal blood flow

Inactive Publication Date: 2008-05-08
STOUT ROBERT L +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0009]The present invention provides a novel approach for determining the presence of drugs in fluid samples. Advantageously, entire families of drugs are identified using the present invention so that one test can provide information on what type of drug is present in a fluid sample. The methods are based on the effects an active drug has on either a target enzyme or a receptor. In the case of drugs which exert their effects on enzymes, the enzymes may be activated or inhibited by the drug binding to the enzyme's catalytic site, thereby inhibiting the binding of the natural substrate. Captopril is a good example of a drug that exhibits this type of competitive inhibition. Captopril is a member of the drug family or class known as Angiotensin Converting Enzyme (ACE) inhibitors which includes the drugs benazepril, captopril, enalapril, fosinopril, lisinopril, quinapril, moexipril, ramipril, and trandolapril. This class or family of drugs assists in regulating blood pressure by inhibiting the conversion of angiotensin I to angiotensin II which is a powerful vasoconstrictor that helps regulate blood pressure, renal blood flow, and blood volume. If there is an excess amount of angiotensin II, which can be caused by the enzymatic action of ACE, blood pressure increases. ACE inhibiting drugs prevent the cleavage of angiotensin I to angiotensin II, thereby reducing blood pressure.
[0011]However, because all members of the ACE-inhibiting drug family act on the same enzyme, the present invention can be used for the detection of the entire family. Advantageously, the active metabolites will also be identified, thereby providing results of the amount of active drug present in a patient's system. Thus, the invention may be used to screen samples for the presence of a class of drugs including their active metabolites. It may also be used to monitor patient compliance or to determine why one drug appears to be more effective in a particular patient. Finally, the present invention will be useful in emergency type situations where it is necessary to quickly ascertain what types of drugs a patient is on, thereby potentially avoiding dangerous drug interactions or needless dosing of additional medications.
[0015]Again, because one test can identify the presence of an active member of a drug class in a sample, time will not have to be spent developing a specific antibody for each member of a drug family. Additionally, information regarding patient compliance with taking medication or efficient detection of active medication in a patient's system are also possible using the present invention.

Problems solved by technology

Such a test does not really provide the needed information of how much active drug is present in a patient's system.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

example 1

[0019]This example tested for the presence of an ACE-inhibiting drug in a fluid sample taken from an individual by measuring enzyme inhibition in the fluid sample.

Materials and Methods

[0020]The synthetic pentapeptide substrate, n-(3(2-furyl)acryloyl)-L-phenylanaylglycylglycine (FAPGG) (Sigma Chemical Company, St Louis, Mo., Cat #305-10) was reconstituted in a bottle with 5 ml of deionized water and left standing for five minutes. The bottle was then inverted a few times and then put on a shaker (Clay Adams CA6000 Centrifuge Becton Dickinson Microbiology System Sparks, Md.) at speed setting 2 for ten minutes. The ACE serum used for this Example was derived from a serum pool consisting of human serum samples which had been tested for ACE activity. However, it is also commercially available through Sigma as part of a kit. All samples, which had an ACE activity greater than 50 units per liter, were combined to form the ACE serum pool. This serum pool was diluted 1:4 with Tris buffer (pH...

example 2

[0026]This example demonstrated that the assay for ACE-inhibiting drugs identified many different medications from the family of ACE-inhibiting drugs.

Materials and Methods

[0027]Fluid samples were obtained from individuals reporting that they were currently taking an ACE-inhibiting drug. The samples and controls were assayed as in Example 1. The medications reported by the patients included eight different medications of the ACE-inhibiting drug family. Each individual reported taking only one specific ACE-inhibiting drug. Thus, this example tests the ability of the assay to identify individuals on ACE-inhibiting drugs without prior knowledge of the specific drug being taken.

Results

[0028]Results from this example are given below in Table 3.

TABLE 3DRUGLISTEDDETECTEDBENAZEPRIL1414CAPTOPRIL22ENALAPRIL55FOSINOPRIL65LISINOPRIL2522MOEXIPRIL43RAMIPRIL22QUINAPRIL1515TOTAL7368

[0029]In this example, 93.1% of urine samples from individuals self-reporting ACE use tested positive for ACE-inhibitin...

example 3

[0030]This example provides a cell receptor assay for B1 adrenergic receptors and tests the accuracy of the assay. Patient urine that may or may not contain target ligand and a radiolabeled competitive ligand are added to a test tube containing a limited concentration of cell membrane containing beta-1-adrenergic receptors. The unlabeled ligand in the patient's urine competes with the labeled ligand for the receptor sites during an incubation period. Following incubation the tubes are centrifuged to precipitate the cell membrane-receptors. The solution containing unbound ligand is decanted and the radioactivity retained in the tubes is detected in a gamma counter. The amount of radioactivity bound is indirectly proportional to the concentration of unlabeled ligand present in the patient's urine.

Materials and Methods

[0031]Tris working buffer (Sigma Chemical) Dissolve 4.55 grams of Tris base, 1.27 g MgCl2 (hexahydrate), 0.37 g disodium dihydrate ethylenediaminetetraacetic acid, and 0....

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PUM

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Abstract

Methods for determining the presence and level of active drugs in fluid samples are provided. Advantageously, entire families or classes of drugs can be tested for in one test by identifying the enzyme or receptor upon which members of that drug family act and measuring enzyme activity levels or binding activity levels of receptors. Methods for establishing standard activity levels of these drugs based upon results from samples having known quantities of drug therein are also provided.

Description

RELATED APPLICATION[0001]This application is a continuation of U.S. patent application Ser. No. 10 / 757,180, filed Jan. 14, 2004, which is a divisional of U.S. patent application Ser. No. 10 / 037,772, filed Nov. 9, 2001, now U.S. Pat. No. 7,094,558, all of which are incorporated by reference herein.BACKGROUND OF THE INVENTION[0002]1. Field of the Invention[0003]The present invention is concerned with the screening for the presence of drugs in fluid samples. More particularly, the present invention is concerned with screening for the presence of drugs in fluid samples which may or may not contain any drugs being screened for. Still more particularly, the present invention is concerned with the screening for the presence of drugs in fluid samples when it is known that the individual providing the fluid sample is on drugs and the screening procedure can help determine proper medication levels. In the instance where it is not known whether or not the individual providing the sample is on ...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): G01N33/53G01N33/566G01N33/94
CPCG01N33/9453G01N33/94
Inventor STOUT, ROBERT L.CHIEN, SUE MINDUNHAM, STEVE H.
Owner STOUT ROBERT L
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