Ex vivo methods for validating substance testing with human organs and/or tissues

• US 20080286747A1
• Filed: 05/18/2007
• Published: 11/20/2008
• Est. Priority Date: 05/18/2007
• Status: Active Grant

• Current Assignee: Lifeline Scientific, Inc. (Shanghai Genext Medical Technology Co., Ltd.)
• Original Assignee: Lifeline Scientific, Inc. (Shanghai Genext Medical Technology Co., Ltd.)
• Inventors: Brassil, John, Kravitz, David, Curtis, Christopher Gerald
• Granted Patent: US 8,765,364 B2
• Time in Patent Office: Days
• US Class Current: 435/1.2
• CPC Class Codes: G01N 33/5088 of vertebrates
• CPC Class Codes: G16H 10/40 for data related to laboratory analysis, e.g. patient specimen analysis
• CPC Class Codes: G16H 20/17 delivered via infusion or injection

Abstract

Methods of validating results of assessments of test substances using human-derived tissues and/or organs, particularly tissues and/or organs unsuitable for transplantation, include assessing the suitability of the organ and/or tissue for substance testing, as well as inter-organ variability and use of exogenous and/or endogenous controls.

Claims

1. A method of validating results of substance testing using at least one human organ or tissue selected from the group consisting of a metabolically active human organ or tissue that has been permanently removed from its origin and an engineered organ or tissue derived from isolated and/or cultured human cells, comprising:

(a) assessing said human organ or tissue for suitability for testing prior to administration of a test substance;

(b) perfusing said organ or tissue with a perfusate containing said test substance;

(c) assessing a parameter relating to said test substance; and

(d) assessing said organ or tissue for suitability for testing of said parameter relating to said test substance during and/or after administration of said test substance.

2. A method of claim 1, wherein step (d) comprises perfusing said organ or tissue with the same perfusate additionally containing at least one control substance having a known qualitative or quantitative effect with respect to said parameter, and assessing the same parameter with respect to at least one said control substance.

3. A method of claim 2, wherein said control substance has a competitive relationship with said test substance relative to said parameter, and is perfused through the organ or tissue after the test substance has been perfused through the organ or tissue.

4. A method of claim 2, wherein said control substance has a competitive relationship with said test substance relative to said parameter, and is perfused through the organ or tissue simultaneously with the test substance.

5. A method of claim 2, wherein said control substance has a non-competitive relationship with said test substance relative to said parameter, and is perfused through the organ or tissue simultaneously with the test substance.

6. A method of claim 2, wherein said at least one control substance is only one control substance.

7. A method of claim 1, wherein step (d) comprises perfusing said organ or tissue with the same perfusate additionally containing at least two control substances having a known qualitative or quantitative effect with respect to said parameter, and assessing the same parameter with respect to at least two said control substances.

8. A method of claim 7, wherein a value of said parameter for a first said control substance has previously been established to be significantly different from a value of said parameter for a second said control substance.

9. A method of claim 8, wherein a value of said parameter for a first said control substance has previously been established to be zero or near zero, and a value of said parameter for a second said control substance has previously been established to be substantially different from zero.

10. A method of claim 2, further comprising determining a ratio between a first value of said parameter for said test substance and a second value of said parameter for said control substance.

11. A method of claim 10, comprising performing said method on a plurality of the same type of organ, establishing a normative range of said ratios, and discounting results of tests in which the ratio is outside of said normative range of ratios.

12. A method of claim 2, wherein said parameter is passive absorption of a substance by organs of the type tested.

13. A method of claim 12, wherein the organs of the type tested are intestines, and the at least one control substance is at least one member selected from the group consisting of antipyrine, terbutaline, mannitol and labeled dextrans.

14. A method of claim 2, wherein said parameter is active absorption of a substance by organs of the type tested, and said at least one control substance is perfused through the organ or tissue after the test substance has been perfused through the organ or tissue.

15. A method of claim 14, wherein the organs of the type tested are intestines, and the at least one control substance is at least one member selected from the group consisting of cephalexin and arginine.

16. A method of claim 14, wherein the organs of the type tested are lungs, and the at least one control substance is salbutamol and ipratopium.

17. A method of claim 2, wherein said parameter is phase I metabolism of a substance by organs of the type tested.

18. A method of claim 17, wherein the organs of the type tested are intestines or kidneys, and the at least one control substance is at least one member selected from the group consisting of phenacetin, tolbutamide, s-methenyltoin, dextromethorphan, chloroxazone and methadone.

19. A method of claim 17, wherein the organs of the type tested are livers, and the at least one control substance is at least one member selected from the group consisting of phenacetin, tolbutamide, s-methenyltoin, dextromethorphan, chloroxazone, methadone, carbomazepine and 7-nitrazepam.

20. A method of claim 17, wherein the organs of the type tested are lungs, and the at least one control substance is at least one member selected from the group consisting of phenacetin, tolbutamide, s-methenyltoin, dextromethorphan, chloroxazone, methadone and carbomazepine.

21. A method of claim 17, comprising selecting said control substance as a function of a type of enzymatic metabolism suspected to occur with said test substance.

22. A method of claim 2, wherein said parameter is phase II metabolism of a substance by organs of the type tested.

23. A method of claim 22, wherein the organs of the type tested are selected from the group consisting of intestines, livers, lungs and kidneys, and the at least one control substance is at least one member selected from the group consisting of harmol and naphthol.

24. A method of claim 8, wherein said parameter is plasma clearance of a substance by livers, and the at least two control substances are propanolol and atenolol.

25. A method of claim 2, wherein said parameter is biliary excretion of a substance by livers, the at least one control substance is at least one member selected from the group consisting of tetrabromosulphophthalein, dibromosulphophthalein, indocyanine green and bile salts, and said at least one control substance is perfused through the organ or tissue after the test substance has been perfused through the organ or tissue.

26. A method of claim 2, wherein said parameter is receptor mediated endocytosis by a substance by livers, and the at least one control substance is at least one member selected from the group consisting of asialoglycoproteins.

27. A method of claim 2, wherein said parameter is active uptake of a substance by lungs, and the at least one control substance is at least one member selected from the group consisting of putrescine, spermine and spermidine, and said at least one control substance is perfused through the organ or tissue after the test substance has been perfused through the organ or tissue.

28. A method of claim 2, wherein said parameter is glomerular filtration of a substance by kidneys, and the at least one control substance is at least one member selected from the group consisting of inulin and creatinine.

29. A method of claim 28, wherein said control substance is radioactively labeled inulin.

30. A method of claim 2, wherein said parameter is active secretion of a substance by kidneys, and the at least one control substance is p-aminohippuric acid, and said at least one control substance is perfused through the organ or tissue after the test substance has been perfused through the organ or tissue.

31. A method of claim 2, wherein said parameter is active reabsorption of a substance by kidneys, and the at least one control substance is tetra-ethylammonium acetate, sodium and glucose, and said at least one control substance is perfused through the organ or tissue after the test substance has been perfused through the organ or tissue.

32. A method of claim 2, wherein said parameter is Phase III metabolism of a substance by kidneys, and the at least one control substance is at least one member selected from the group consisting of mercaptans and glutathione conjugates.

33. A method of claim 8, wherein said parameter is uptake of a substance by hearts, and the at least one control substance is dopa and dopamine.

34. A method of claim 2, wherein said control substance is an endogenous substance.

35. A method of claim 2, wherein said control substance is an exogenous substance.

36. A method of claim 2, wherein said perfusing steps are carried out under normothermic conditions.

37. A method of claim 1, wherein said organ or tissue has been subjected to hypothermic storage before step (a).

38. A method of claim 1, wherein said organ or tissue has been subjected to disease or injury before step (a).

39. A method of claim 38, wherein said disease or injury occurred before harvest of said organ or tissue.

40. A method of claim 38, wherein said disease or injury occurred after harvest of said organ or tissue.

41. A method of claim 1, wherein said organ or tissue has been subjected to injury before step (a), and said injury resulted from prolonged warm ischemia.

42. A method of claim 1, wherein said organ or tissue has been subjected to injury before step (a), and said injury resulted from hypothermic storage.

43. A method of claim 1, wherein said at least one organ or tissue is unsuitable for transplant.

44. A method of claim 1, wherein said at least one organ or tissue is a combination of organs.

45. A method of claim 1, wherein said at least one organ or tissue is an organ or tissue harvested from a deceased human.

46. A method of claim 1, wherein said at least one organ or tissue is an engineered organ or tissue.

47. A method of claim 1, wherein said parameter is assessed in at least one member selected from the group consisting of a perfusate sample, a biopsy from said at least one organ or tissue, and an effluent of said at least one organ or tissue.

48. A method of generating revenue, comprising:

charging a fee to a third party for collecting data relating to effects, properties and or fate of a test substance when administered to an organ or tissue;

performing the method of claim 1; and

presenting results assessed in step (d) along with results of said substance testing to said third party.