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Generated: August 20, 2019

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Claims for Patent: 7,897,363

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Summary for Patent: 7,897,363
Title:Methods of screening an agent for an activity in an isolated eye of a teleost
Abstract: The present invention provides methods of screening an agent for an activity in an isolated organ, e.g., eye, from a teleost, e.g., zebrafish. Methods of isolating eyes from zebrafish are provided. Methods of screening an agent for an ocular activity in the isolated eye are provided. Methods of screening an agent for an ocular activity in a model of ocular disease or disorder are provided. Methods of screening an agent for an ocular activity in the isolated eye and for screening the agent for cell death and/or toxic activity in the eye or other organ or tissue are provided. The invention further provides high throughput methods of screening agents for an activity in isolated eyes of zebrafish in multi-well plates.
Inventor(s): McGrath; Patricia (Cambridge, MA), Seng; Wen Lin (Westborough, MA)
Assignee: Phylonix Pharmaceuticals, Inc. (Cambridge, MA)
Application Number:12/137,509
Patent Claims:1. A method of analyzing an eye from a teleost, the method comprising: contacting the teleost with an enzyme that dissociates the eye from the teleost, wherein the enzyme is a collagenase, a dispase, a trypsin, a chymotrypsin, or a hyaluronidase; and analyzing the dissociated eye.

2. The method of claim 1, further comprising contacting the teleost with an agent, and analyzing a response to the agent in the dissociated eye.

3. The method of claim 1, further comprising collecting the dissociated eye by filtration or density gradient centrifugation.

4. The method of claim 1, wherein the enzyme is a collagenase.

5. The method of claim 1, wherein the teleost is in a well of a multi-well plate.

6. The method of claim 5, further comprising removing the teleost body from the well before the analyzing step.

7. The method of any claim 2, wherein the agent is an inducer or potential inducer of ocular disease.

8. The method of claim 7, wherein the response indicates whether the inducer or potential inducer induces the disease.

9. The method of claim 2, wherein the teleost has or is susceptible to an ocular disease and the agent is an inhibitor or potential inhibitor of ocular disease.

10. The method of claim 2, wherein first and second agents are administered, the first agent being an inducer of ocular disease and the second agent being an inhibitor or potential inhibitor of the disease, and the analyzed response provides an indication whether the second agent inhibits the disease, wherein the first and second agents can be administered in either order or together.

11. The method of claim 2, wherein the response is analyzed using a microplate reader, a high content imaging system, or a microscope.

12. The method of claim 1, wherein the isolated eye is contained on a slide following isolation of the eye from the teleost.

13. The method of claim 2, wherein the analyzing step comprises detecting a target biomolecule.

14. The method of claim 13, wherein the target biomolecule is a protein or mRNA.

15. The method of claim 2, wherein the analyzing step is performed on the isolated eye in situ.

16. The method of claim 2, wherein the response to the agent comprises an increase or decrease in angiogenesis.

17. The method of claim 16, wherein the response to the agent comprises an increase or decrease in blood vessel formation.

18. The method of claim 17, wherein the blood vessels are visualized by staining of the eye with a vessel-specific antibody.

19. The method of claim 1, wherein the teleost is an embryo, larva, or adult.

20. The method of claim 1 , wherein the teleost is a zebrafish, medaka, Giant rerio, or puffer fish.

21. The method of claim 1, wherein the teleost is a wildtype teleost.

22. The method of claim 1, wherein the teleost is mutant or transgenic teleost.

23. The method of claim 2, wherein the agent is administered to the teleost by dissolving the agent in media used for culturing the teleost.

24. The method of claim 2, wherein the agent is administered to the teleost by injecting the agent into the teleost.

25. The method of claim 2, wherein the agent is a small molecule, nucleic acid, nucleic acid analog, peptide, protein, glycoprotein, carbohydrate, lipid, or glycolipid.

26. The method of claim 2, wherein the agent is a member of a library of agents is screened for an ocular activity in the isolated eyes of a plurality of teleosts.

27. The method of claim 2, wherein analyzing the response to the agent comprises assessing the isolated eye for a morphological change.

28. The method of claim 27, wherein the morphological change comprises a change in size, shape, pigmentation, color, or structure of the eye.

29. The method of claim 27, wherein the morphological change comprises a change in blood vessel structure.

30. The method of claim 27, wherein assessing the morphological change comprises acquiring an image of the isolated eye.

31. The method of claim 30, wherein the acquired image is a digital image and assessment of the morphological change comprises computer-based analysis of the digital image.

32. The method of claim 2, wherein the agent induces ocular neovascularization in the teleost.

33. The method of claim 32, wherein the agent that induces ocular neovascularization in the teleost is CoCl.sub.2 or Penicillamine.

34. The method of claim 2, further comprising: surgically cutting the conjunctiva and then burning the sclera of the teleost to induce ocular scarring; and wherein the analyzing step comprises determining whether the agent reduce scarring of the sclera relative to a control teleost not treated with the agent; wherein the cutting step and the administering step can be performed in either order or together.

Details for Patent 7,897,363

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 Phylonix Pharmaceuticals, Inc. (Cambridge, MA) 2036-04-30 RX search
>Applicant >Tradename >Biologic Ingredient >Dosage Form >BLA >Number >Approval Date >Patent No. >Assignee >Estimated Patent Expiration >Status >Orphan >Source

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International Patent Family for US Patent 7,897,363

Country Patent Number Publication Date
World Intellectual Property Organization (WIPO) 2008154641 Dec 18, 2008
United States of America 2009010849 Jan 08, 2009
United States of America 2011218119 Sep 08, 2011
United States of America 2013281320 Oct 24, 2013
United States of America 8367364 Feb 05, 2013
United States of America 8741592 Jun 03, 2014
>Country >Patent Number >Publication Date

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