Make Better Decisions - Finding and Evaluating Generic and Branded Drug Market Entry Opportunities

Get the Book: Make Better Decisions

Finding and Evaluating Generic and Branded Drug Market Entry Opportunities

PDF eBook: Just $10 Get Print Book on Amazon

Serving hundreds of leading biopharmaceutical companies globally:

Teva
Deloitte
US Department of Justice
Covington
Moodys
Merck

Generated: August 26, 2019

DrugPatentWatch Database Preview

Claims for Patent: 5,814,460

  Try a free trial


See Plans and Pricing

« Back to Dashboard

Summary for Patent: 5,814,460
Title: Method for generating and screening useful peptides
Abstract:The invention allows the generation and screening of a large population of peptides for the presence of peptides which bind a particular macromolecule or macromolecular complex with high affinity, and further allows the favored net synthesis of analyzable quantities of such peptides, by using is the \"trap\" a macromolecule or macromolecular complex for which binding of the peptide is desired. The starting mixture is preferably spiked with a peptide having some affinity for the target macromolecule so that mutation of the spike or \"lead\" peptide is favored. The development of improved binding peptides through scrambling may be dynamically monitored by initially binding the target with an insolubilized ligand, and then looking for an increase in the concentration of the target in the soluble phase as a result of the displacement of the reference ligand by scrambled peptides.
Inventor(s): Venton; Duane L. (Lombard, IL), Hopfinger; Anton J. (Lake Forest, IL), Lebreton; Guy (Oak Park, IL)
Assignee: Diatide, Inc. (Londonderry, NH)
Application Number:08/387,749
Patent Claims:1. A method of identifying peptides which bind specifically to a predetermined target, comprising the steps of:

(a) subjecting a mixture initially comprising a starting protein and/or plurality of starting peptides to conditions, comprising proteolytic enzyme exposure, under which the starting proteins and/or peptides and derivatives thereof can undergo both random degradation into smaller peptide and free amino acid derivatives, and random recombination of the starting proteins and/or peptides, and/or their derivatives, into new peptides, whereby the component amino acids of the starting mixture are scrambled to generate a diverse population of scrambled peptides of different sequences, said scrambled peptides not being characterized by a predetermined amino acid sequence;

(b) allowing the starting peptides, if any, peptide fragments of the starting peptides and/or proteins, and scrambled peptides, to compete with each ocher to bind to the target to form a specifically bound peptide-target complex;

(c) protecting only those competing peptides which are bound to the target from further scrambling;

(d) recovering the specifically bound peptide from a peptide-target complex;

(e) sequencing a recovered specifically bound peptide, wherein the improvement comprises monitoring the formation of peptide-target complex, said monitoring effected directly or indirectly, intermittently or continuously, during the scrambling reaction, by a monitoring method comprising incubating the target with a labeled ligand specific for the target, which ligand is disclaceable by one or more scrambled peptides, and measuring the concentration of free-labeled ligand, target-bound ligand, or unlabeled reagent.

2. The method of claim 1 in which both the ligand and the target are soluble.

3. The method of claim 2 in which, in step (e), free labeled ligand, target bound ligand, and unlabeled target are chromatographically separated.

4. The method of claim 2 in which free labeled ligand is measured.

5. The method of claim 2 in which targeted bound ligand is measured.

6. The method of claim 2 in which unlabeled target is measured.

7. The method of claim 2 in which the label is fluorescent and the measurement is spectroscopic and in situ.

8. The method of claim 1 in which the ligand is immobilized.

9. The method of claim 1 in which the receptor is immobilized.

10. The method of claim 1 wherein the proteolytic enzyme is selected from the group consisting of papain, pepsin, bromelin, thermolysin, trypsin, pronase, chymotrypsin, subtilisin and dipeptidyl peptidase IV.

11. The method according to claim 1 wherein the protection from further scrambling is provided by interposing a semipermeable membrane between said enzymes and said target, said membrane being permeable to said peptides and impermeable to said target and said enzymes.

12. The method of claim 11 wherein the semipermeable membrane has a permeability cutoff of about 1-3.5 kDa.

13. the method of claim 1 wherein protection from further scrambling is provided by immobilizing the enzymes in a first zone and the targets in a second, spatially separated zone, the unbound peptides generated by the scrambling reaction being allowed to circulate between the first and second zone.

14. The method of claim 1, further comprising monitoring the peptide size distribution from time to time while the scrambling reaction is in progress.

15. The method of claim 14, further comprising adjusting the peptide size distribution during the course of the scrambling reaction by adding more amino acids, peptides or protein, or by diluting the reaction mixture.

16. The method claim 1 wherein the average length of the peptides of the starting mixture is in the range of 7 to 10 amino acids.

17. The method of claim 1 wherein substantially all of the twenty genetically encoded amino acids are represented in the peptides of the starting mixture.

18. The method of claim 1, wherein the starting mixture is biased in favor of or against certain predetermined amino acids.

19. The method of claim 1, wherein the starting mixture is spiked with a peptide of known sequence and having an affinity of at least about 10.sup.-4 for the target.

20. The method according to claim 1 wherein the specific target is a receptor involved in a physiological process.

21. The method of claim 1, wherein the scrambled peptides are simultaneously screened for affinity for each of a plurality of different targets.

22. The method of claim 1 wherein the target is a macromolecule or a macromolecular complex.

23. The method according to claim 22 wherein the macromolecule or macromolecular complex is fibrinogen, sickle cell hemoglobin, collagenase IV, renin, GpII.sub.b III.sub.a or phospholipase A.sub.2.

24. The method of claim 11 wherein said semipermeable membrane defines a scrambling zone and a binding zone, the target is provided in soluble form but cannot pass the membrane, and the binding of the scrambled peptides to the predetermined target in the binding zone is monitored by:

(i) incubating an insolubilized ligand having a known affinity for the target with said target, prior to commencing the scrambling of step (a), and

(ii) whenever it is desired to monitor the binding of the scrambled peptides to the target, sampling the soluble fraction from binding zone and assaying the sample for the presence of the soluble target.

Summary for Patent: ➤ Sign Up

PCT Information
PCT FiledAugust 09, 1993PCT Application Number:PCT/US93/08231
PCT Publication Date:March 03, 1994PCT Publication Number:WO94/04558

Details for Patent 5,814,460

Applicant Tradename Biologic Ingredient Dosage Form BLA Number Approval Date Patent No. Assignee Estimated Patent Expiration Status Orphan Source
Advance Biofactures SANTYL collagenase VIAL 101995 001 1965-06-04 ➤ Sign Up Diatide, Inc. (Londonderry, NH) 2015-09-29 RX search
>Applicant >Tradename >Biologic Ingredient >Dosage Form >BLA >Number >Approval Date >Patent No. >Assignee >Estimated Patent Expiration >Status >Orphan >Source

Subscribe to access the full database, or try a Free Trial

International Patent Family for US Patent 5,814,460

Country Patent Number Publication Date
World Intellectual Property Organization (WIPO) 9112331 Aug 22, 1991
World Intellectual Property Organization (WIPO) 9304079 Mar 04, 1993
World Intellectual Property Organization (WIPO) 9404558 Mar 03, 1994
United States of America 5366862 Nov 22, 1994
Japan H05506774 Oct 07, 1993
Japan H07502111 Mar 02, 1995
Israel 102897 Jan 31, 1993
>Country >Patent Number >Publication Date

Subscribe to access the full database, or try a Free Trial

Make Better Decisions: Try a trial or see plans & pricing

Serving hundreds of leading biopharmaceutical companies globally:

Harvard Business School
Colorcon
Express Scripts
Fuji
Cantor Fitzgerald
Chubb

Drugs may be covered by multiple patents or regulatory protections. All trademarks and applicant names are the property of their respective owners or licensors. Although great care is taken in the proper and correct provision of this service, thinkBiotech LLC does not accept any responsibility for possible consequences of errors or omissions in the provided data. The data presented herein is for information purposes only. There is no warranty that the data contained herein is error free. thinkBiotech performs no independent verifification of facts as provided by public sources nor are attempts made to provide legal or investing advice. Any reliance on data provided herein is done solely at the discretion of the user. Users of this service are advised to seek professional advice and independent confirmation before considering acting on any of the provided information. thinkBiotech LLC reserves the right to amend, extend or withdraw any part or all of the offered service without notice.