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Details for Patent: 6,048,514

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Details for Patent: 6,048,514

Title: Amylin activity assays
Abstract:Novel methods for use in identifying or assaying compounds which can simulate the ability of amylin to cause hyperlactemia and hyperglycemia in in vivo biological models, or for use in evaluating the potency of compounds known or suspected to simulate these actions of amylin, which involve introducing test samples into in vivo test systems and determining the presence or amount of a rise in lactate, or determining the presence or amount of a rise in lactate and a rise in glucose, following test sample administration.
Inventor(s): Young; Andrew A. (San Diego, CA), Cooper; Garth J. S. (Solana Beach, CA), Rink; Timothy J. (La Jolla, CA)
Assignee: Amylin Pharmaceuticals, Inc. (San Diego, CA)
Filing Date:Apr 14, 1995
Application Number:08/422,747
Claims:1. An assay method for use in identifying a test compound which can simulate the activity of amylin which method comprises:

a. bringing together a test sample and a test system, said test sample comprising one or more test compounds, and said test system comprising an in vivo biological model, said in vivo model being characterized in that it exhibits elevated lactate and elevated glucose in response to the introduction to said model of amylin or an amylin agonist;

b. determining the presence or amount of a rise in lactate and the presence or amount of a rise in glucose in said test system;

c. determining whether a peak in elevated lactate preceded a peak in elevated glucose; and

d. identifying those test compounds which resulted in a peak in elevated lactate which preceded a peak in elevated glucose in the in vivo biological model in which at least one test compound in the test sample brought together with the test system results in a peak in elevated lactate which precedes a peak in elevated glucose.

2. The method of claim 1 which further comprises the use of a positive control assay, a negative control assay, or both.

3. The method of claim 2 wherein said positive control assay is carried out according to the method of claim 1 and said test sample comprises amylin or an amylin agonist.

4. The method of claim 1 wherein said in vivo model is a rat.

5. The method of claim 4 wherein said method is carried out using more than one rat, said rats being fed, or fasted, or both.

6. An assay method for use in evaluating the potency of a test compound known or suspected to simulate the activity of amylin, which method comprises:

a. bringing together a test sample and a test system, said test sample comprising one or more test compounds, and said test system comprising an in vivo biological model, said in vivo model being characterized in that it exhibits elevated lactate and elevated glucose in response to the introduction to said model of amylin or an amylin agonist;

b. determining the presence or amount of a rise in lactate and the presence or amount of a rise in glucose in said test system;

c. determining whether a peak in elevated lactate preceded a peak in elevated glucose; and

d. evaluating the potency of those test compounds which resulted in a peak in elevated lactate which preceded a peak in elevated glucose in the in vivo biological model in which at least one test compound in the test sample brought together with the test system results in a peak in elevated lactate which precedes a peak in elevated glucose.

7. The method of claim 6, which further comprises the steps of repeating said assay method using differing amounts of said test sample, and generating a dose response profile for said rise in lactate for use in evaluating the potency of said test sample in simulating the activity of amylin.

8. The method of claim 6, which further comprises the steps of repeating said assay method using differing amounts of said test sample, and generating a dose response profile for said rise in glucose for use in evaluating the potency of said test sample in simulating the activity of amylin.

9. The method of claim 6, which further comprises the steps of repeating said assay method using differing amounts of said test sample, and generating a dose response profile for said rise in glucose and for said rise in lactate for use in evaluating the potency of said test sample in simulating the activity of amylin.

10. The method of claim 7, which further comprises comparing said dose response profile for said test sample to a dose response profile for said test sample to a dose response profile prepared for one or more positive control assays, or one or more negative control assays, or both, and evaluating the potency of said test sample in simulating the activity of amylin.

11. The method of claim 8, which further comprises comparing said dose response profile for said test sample to a dose response profile for said test sample to a dose response profile prepared for one or more positive control assays, or one or more negative control assays, or both, and evaluating the potency of said test sample in simulating the activity of amylin.

12. The method of claim 9, which further comprises comparing said dose response profile for said test sample to a dose response profile prepared for one or more positive control assays, or one or more negative control assays, or both, and evaluating the potency of said test sample in simulating the activity of amylin.
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