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Last Updated: March 29, 2024

Claims for Patent: 8,367,364


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Summary for Patent: 8,367,364
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:13/037,104
Patent Claims:1. A method of screening an agent for an ocular or ophthalmologic activity in 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; isolating the eye; contacting the eye with an agent to be tested for the ocular or ophthalmologic activity; and analyzing a response to the agent in the dissociated eye, wherein the presence of the response is indicative of the ocular or ophthalmologic activity of the agent.

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

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

4. The method of claim 1, wherein the teleost has or is susceptible to an ocular disease and the agent is an inhibitor or potential inhibitor of ocular disease.

5. The method of claim 1, 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.

6. The method of claim 1, wherein the response is analyzed using a microplate reader, a high content imaging system, or a microscope.

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

8. The method of claim 1, wherein the analyzing step comprises detecting a target biomolecule.

9. The method of claim 8, wherein the target biomolecule is a protein or mRNA.

10. The method of claim 1, wherein the analyzing step is performed on the isolated eye in situ.

11. The method of claim 1, wherein the response to the agent comprises an increase or decrease in angiogenesis.

12. The method of claim 11, wherein the response to the agent comprises an increase or decrease in blood vessel formation.

13. The method of claim 11, wherein the blood vessels are visualized by staining of the eye with a vessel-specific antibody.

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

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

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

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

18. The method of claim 1, wherein the agent is administered to the teleost by dissolving the agent in media used for culturing the teleost.

19. The method of claim 1, wherein the agent is administered to the teleost by injecting the agent into the teleost.

20. The method of claim 1, wherein the agent is a small molecule, nucleic acid, nucleic acid analog, peptide, protein, glycoprotein, carbohydrate, lipid, or glycolipid.

21. The method of claim 1, wherein the agent is a member of a library of agents is screened for an ocular or ophthalmologic activity in the isolated eyes of a plurality of teleosts.

22. The method of claim 1, wherein analyzing the response to the agent comprises assessing the isolated eye for a morphological change.

23. The method of claim 22, wherein the morphological change comprises of a change in size, shape, pigmentation, color, or structure of the eye.

24. The method of claim 22, wherein the morphological change comprises a change in blood vessel structure.

25. The method of 22, wherein the assessing the morphological change comprises acquiring an image of the isolated eye.

26. The method of claim 25, wherein the acquired image is a digital image and assessment of the morphological change comprises computer-based analysis of the digital image.

27. The method of claim 1, wherein the agent induces ocular or choroidal neovascularization in the teleost.

28. The method of claim 27, wherein the agent that induces ocular neovascularization in the teleost is CoCl.sub.2 or Penicillamine.

Details for Patent 8,367,364

Applicant Tradename Biologic Ingredient Dosage Form BLA Approval Date Patent No. Expiredate
Bausch & Lomb Incorporated VITRASE hyaluronidase Injection 021640 05/05/2004 ⤷  Try a Trial 2027-06-12
Bausch & Lomb Incorporated VITRASE hyaluronidase Injection 021640 12/02/2004 ⤷  Try a Trial 2027-06-12
Amphastar Pharmaceuticals, Inc. AMPHADASE hyaluronidase Injection 021665 10/26/2004 ⤷  Try a Trial 2027-06-12
Akorn, Inc. HYDASE hyaluronidase Injection 021716 10/25/2005 ⤷  Try a Trial 2027-06-12
Smith & Nephew, Inc. SANTYL collagenase Ointment 101995 06/04/1965 ⤷  Try a Trial 2027-06-12
>Applicant >Tradename >Biologic Ingredient >Dosage Form >BLA >Approval Date >Patent No. >Expiredate

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