Claims for Patent: 8,741,592

Summary for Patent: 8,741,592

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:	13/734,271
Patent Claims:	1. A method of screening an agent for a protective or therapeutic activity for an ocular disease, the method comprising: contacting a teleost with a first agent that is an inducer of ocular disease, and a second agent that is a protective or therapeutic agent for the disease; 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 of the teleost; and analyzing a response to the second agent in the dissociated eye, wherein the presence of the response is indicative of the protective or therapeutic 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 first and second agents can be administered in either order or together. 5. The method of claim 1, wherein the response is analyzed using a microplate reader, a high content imaging system, or a microscope. 6. The method of claim 1, wherein the isolated eye is contained on a slide following isolation of the eye from the teleost. 7. The method of claim 1, wherein the analyzing step comprises detecting a target biomolecule. 8. The method of claim 7, wherein the target biomolecule is a protein or mRNA. 9. The method of claim 1, wherein the analyzing step is performed on the isolated eye in situ. 10. The method of claim 1, wherein the response to the agent comprises an increase or decrease in angiogenesis. 11. The method of claim 10, wherein the response to the agent comprises an increase or decrease in blood vessel formation. 12. The method of claim 10, wherein the blood vessels are visualized by staining of the eye with a vessel-specific antibody. 13. The method of claim 1, wherein the teleost is an embryo, larva, or adult. 14. The method of claim 1, wherein the teleost is a zebrafish, medaka, Giant rerio, or puffer fish. 15. The method of claim 1, wherein the teleost is a wildtype teleost. 16. The method of claim 1, wherein the teleost is a mutant or transgenic teleost. 17. The method of claim 1, wherein the first or the second agent, or both, is administered to the teleost by dissolving the agent in media used for culturing the teleost. 18. The method of claim 1, wherein the first or the second agent, or both, is administered to the teleost by injecting the agent into the teleost. 19. The method of claim 1, wherein the agent is a small molecule, nucleic acid, nucleic acid analog, peptide, protein, glycoprotein, carbohydrate, lipid, or glycolipid. 20. The method of claim 1, wherein the second agent is a member of a library of agents is screened for protective or therapeutic activity in the isolated eyes of a plurality of teleosts. 21. The method of claim 1, wherein analyzing the response to the agent comprises assessing the isolated eye for a morphological change. 22. The method of claim 21, wherein the morphological change comprises of a change in size, shape, pigmentation, color, or structure of the eye. 23. The method of claim 21, wherein the morphological change comprises a change in blood vessel structure. 24. The method of 21, wherein the assessing the morphological change comprises acquiring an image of the isolated eye. 25. The method of claim 24, wherein the acquired image is a digital image and assessment of the morphological change comprises computer-based analysis of the digital image. 26. A method of screening an agent for a protective or therapeutic activity for ocular scarring, the method comprising: surgically cutting conjunctiva and then burning the sclera of a teleost to include ocular scarring; contacting the teleost with an agent that is a potential protective or therapeutic agent; 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 of the teleost; and analyzing a response to the agent in the dissociated eye, wherein the presence of reduced scarring of the sclera in the presence of the agent, relative to a control teleost not treated with the agent, is indicative of the protective or therapeutic activity of the agent. 27. The method of claim 26, wherein the cutting of the conjunctiva and the contacting with the agent are performed in either order, or together.

Details for Patent 8,741,592

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