Peritoneal therapeutic fluid

Abstract

Peritoneal therapeutic fluid comprising one or more of a biocompatibility enhancing agent (BCA) that is selected from the group consisting of a polyphenolic compound, a metabolite of a polyphenolic compound which is obtained by metabolization in the human or animal body, a salt or a glycoside of a polyphenolic compound.

Claims

1. A method comprising administering to a human patient a peritoneal therapeutic fluid, the peritoneal therapeutic fluid comprising at least one biocompatibility enhancing agent (BCA), wherein the BCA is resveratrol, dihydro-resveratrol, piceid, piceatannol, pterostilbene, piceid glucoside, caffeic acid, luteolin, or a salt thereof, to conduct peritoneal dialysis on the human patient, wherein cytotoxicity on human peritoneal mesothelial cells is decreased compared to other peritoneal therapeutic fluids that do not contain the BCA, wherein the at least one BCA is present in a concentration of between 0.05 to 20 μMol/L, and wherein the peritoneal therapeutic fluid comprises electrolytes.

2. The method according to claim 1, wherein the peritoneal therapeutic fluid is an aqueous solution comprising: sodium in an amount of 90 to 150 mEq/L; potassium in an amount of 0 to 5 mEq/L; calcium in an amount of 0 to 6 mEq/L; magnesium in an amount of 0 to 4 mEq/L; and an alkali equivalent in an amount of 25 to 50 mEq/L.

3. The method according to claim 1, wherein the peritoneal therapeutic fluid further comprises at least one saccharide.

4. The method according to claim 1, further comprising at least one disaccharide selected from the group consisting of sucrose, Gentiobiulose, Laminaribiose, Gentiobiose, Rutinulose, Xylobiose, trehalose, β,β-Trehalose, α,β-Trehalose, lactulose, sophorose, lactose, cellobiose, chitobiose, maltose, Kojibiose, Nigerose, Isomaltose, Turanose, Maltulose, Palatinose, Mannobiose, Melibiose, Melibiulose, and Rutinose.

5. The method according to claim 1, wherein the peritoneal therapeutic fluid further comprises maltodextrin or at least one oligosaccharide that is a product of limited hydrolysis of one of more of the following: starch, amylose, amylopectin, fructan, glucan, galactan, mannan, cellulose, arabic gum, glycogen, dextran, hemicellulose, arabinoxylose, and pectin, wherein the product of limited hydrolysis has a molecular weight of 90 D to 500 kD.

6. The method according to claim 1, wherein the peritoneal therapeutic fluid further comprises at least one alpha-glucan with a degree of polymerization of 3 or higher and has a molecular weight of 90 D to 500 kD.

7. The method according to claim 1, wherein the peritoneal therapeutic fluid further comprises at least one saccharide which is selected from the group consisting of isomaltotriose, nigerotriose, maltotriose, melezitose, maltotriulose, raffinose, kestose, maltodextrin, dextrins, heparin, Dextran, glycogen, pullulan, starch, amylose, amylopectin, icodextrin, and combinations thereof.

8. The method according to claim 1, wherein the peritoneal therapeutic fluid further comprises at least one saccharide which has a molecular weight in a range of 90D to 50 kD, 90D to 500 D, 90D to 1.5 kD, 1.5 kD to 50 kD, 350D to 50 kD, 250D and 50 KD, or 150 to 400 D.

9. The method according to claim 1, wherein the peritoneal therapeutic fluid further comprises at least one saccharide, and wherein the at least one saccharide is present in a total concentration of ≤0.02% by weight (200 mg/L), ≤0.75% by weight, ≤2.4% by weight, ≤5% by weight, ≤7.5% by weight, or ≤20% by weight.

10. The method according to claim 1, wherein the peritoneal therapeutic fluid is used for decreasing expression of Vascular Endothelial Growth Factor (VEGF) in the peritoneum.

11. The method according to claim 1, wherein the peritoneal therapeutic fluid is used for decreasing long term fibrosis.

12. The method according to claim 1, wherein the biocompatibility enhancing agent is pegylated with Polyethyleneglycol (PEG) or Methoxy-Polyethyleneglycol (mPEG), provided that the BCA is not polyethylene glycol (PEG) or a derivative of a polyethylene glycol.

13. The method according to claim 1, wherein the peritoneal therapeutic fluid further comprises at least one ingredient which is selected from the group consisting of alkali metal ions, alkaline earth metal ions, an osmotic agent and a pH-buffer.

14. The method according to claim 1, wherein the peritoneal therapeutic fluid further comprises at least one osmotic agent.

15. The method according to claim 1, wherein the peritoneal therapeutic fluid further comprises glucose.

16. The method according to claim 1, wherein the peritoneal therapeutic fluid further comprises a saccharide selected from the group consisting of fructose, a disaccharide, an oligosaccharide, maltodextrin and a polysaccharide, or any combination thereof.

17. The method according to claim 1, wherein the biocompatibility enhancing agent is resveratrol.

18. The method according to claim 1, wherein the peritoneal therapeutic fluid is an aqueous solution has an alkali equivalent in an amount of 25 to 50 mEq/L.

19. The method according to claim 1, wherein the electrolytes comprise one or more of sodium, potassium, calcium, and/or magnesium.

20. A method comprising administering to a human patient a peritoneal therapeutic fluid, the peritoneal therapeutic fluid comprising at least one biocompatibility enhancing agent (BCA), wherein the BCA is a resveratrol derivative, to conduct peritoneal dialysis on the human patient, wherein cytotoxicity on human peritoneal mesothelial cells is decreased compared to other peritoneal therapeutic fluids that do not contain the BCA, wherein the at least one BCA is present in a concentration of between 0.05 to 20 μMol/L, wherein the peritoneal therapeutic fluid comprises electrolytes, and wherein the resveratrol derivative is a compound of formula 2 or 8: ##STR00021## ##STR00022## ##STR00023## wherein in compound 2 R1=R2=R4=OCH3, R3=R5=R6=H; or R1=R2=R3=R5=OCH3, R4=R6=H; or R1=R2=R3=R4=OCH3, R5=R6=H, and wherein in compound 8 R1=OCH3, R2=OH, R3=O-Glucose; or R1=OCH3, R2=H, R3=O-Glucose; or R1=OCH3, R2=OH, R3=OH; or R1=OCH3, R2=H, R3=OH; or R1=OH, R2=OH, R3=O-Glucose.

21. A method comprising administering to a human patient a peritoneal therapeutic fluid, the peritoneal therapeutic fluid comprising at least one biocompatibility enhancing agent (BCA), wherein the BCA is a resveratrol derivative, to conduct peritoneal dialysis on the human patient, wherein cytotoxicity on human peritoneal mesothelial cells is decreased compared to other peritoneal therapeutic fluids that do not contain the BCA, wherein the at least one BCA is present in a concentration of between 0.05 to 20 μMol/L, wherein the peritoneal therapeutic fluid comprises electrolytes, and wherein the resveratrol derivative is a compound of formula 19: ##STR00024## wherein R4 is ##STR00025## wherein R1, R2, R3, R11, R12, R13, R14, and R15 are independently from each other selected from —H, —OH, —O—R.sub.Alk, —CHO, —COR.sub.Alk, —COOH, —COO—R.sub.Alk, —CO—NH—C.sub.nH.sub.2n—COOH, —CO—NH—C.sub.2nH.sub.2n—COO, —CN, —Cl, —Br, —I, —NO.sub.2, —C.sub.nH.sub.2nCN, —C.sub.nH.sub.2n—Cl, —C.sub.nH.sub.2n—Br, —C.sub.nH.sub.2n—I, —C.sub.nH.sub.2n—NO.sub.2, —O—PO.sub.3.sup.2−, —O—PO.sub.3H.sup.−, —O—PO.sub.3H.sub.2, —NH.sub.2, —NHR.sub.Alk, —NR.sub.Alk1R.sub.Alk2, —N.sup.+H.sub.3, —N.sup.+H.sub.2R.sub.Alk, —N.sup.+HR.sub.Alk1,R.sub.Alk2, —N.sup.+ R.sub.Alk1,R.sub.Alk2R.sub.Alk3, —B(OH).sub.2, —OCHO, —O—COR.sub.Alk, —OCF.sub.3, —O—CN, —OCH.sub.2CN, wherein R.sub.Alk, R.sub.Alk1, R.sub.Alk2, and R.sub.Alk3 are alkyl residues which are independently selected from each other, wherein the alky residues are CH.sub.3, C.sub.2H.sub.5, C.sub.3H.sub.7 or C.sub.4H.sub.9, and wherein C.sub.nH.sub.2n is CH.sub.2, C.sub.2H.sub.4, C.sub.3H.sub.6, or C.sub.4H.sub.8.

22. The method according to claim 20, wherein the peritoneal therapeutic fluid is an aqueous solution comprising: sodium in an amount of 90 to 150 mEq/L; potassium in an amount of 0 to 5 mEq/L; calcium in an amount of 0 to 6 mEq/L; magnesium in an amount of 0 to 4 mEq/L; and an alkali equivalent in an amount of 25 to 50 mEq/L.

23. The method according to claim 20, wherein the peritoneal therapeutic fluid further comprises at least one saccharide.

24. The method according to claim 20, further comprising at least one disaccharide selected from the group consisting of sucrose, Gentiobiulose, Laminaribiose, Gentiobiose, Rutinulose, Xylobiose, trehalose, β,β-Trehalose, α,β-Trehalose, lactulose, sophorose, lactose, cellobiose, chitobiose, maltose, Kojibiose, Nigerose, Isomaltose, Turanose, Maltulose, Palatinose, Mannobiose, Melibiose, Melibiulose, and Rutinose.

25. The method according to claim 20, wherein the peritoneal therapeutic fluid further comprises maltodextrin or at least one oligosaccharide that is a product of limited hydrolysis of one of more of the following: starch, amylose, amylopectin, fructan, glucan, galactan, mannan, cellulose, arabic gum, glycogen, dextran, hemicellulose, arabinoxylose, and pectin, wherein the product of limited hydrolysis has a molecular weight of 90 D to 500 kD.

26. The method according to claim 20, wherein the peritoneal therapeutic fluid further comprises at least one alpha-glucan with a degree of polymerization of 3 or higher and has a molecular weight of 90 D to 500 kD.

27. The method according to claim 20, wherein the peritoneal therapeutic fluid further comprises at least one saccharide which is selected from the group consisting of isomaltotriose, nigerotriose, maltotriose, melezitose, maltotriulose, raffinose, kestose, maltodextrin, dextrins, heparin, Dextran, glycogen, pullulan, starch, amylose, amylopectin, icodextrin, and combinations thereof.

28. The method according to claim 20, wherein the peritoneal therapeutic fluid further comprises at least one saccharide which has a molecular weight in a range of 90D to 50 kD, 90D to 500 D, 90D to 1.5 kD, 1.5 kD to 50 kD, 350D to 50 kD, 250D and 50 KD, or 150 to 400 D.

29. The method according to claim 20, wherein the peritoneal therapeutic fluid further comprises at least one saccharide, and wherein the at least one saccharide is present in a total concentration of ≥0.02% by weight (200 mg/L), ≥0.75% by weight, ≥2.4% by weight, ≥5% by weight, ≥7.5% by weight, or ≥20% by weight.

30. The method according to claim 20, wherein the peritoneal therapeutic fluid is used for decreasing expression of Vascular Endothelial Growth Factor (VEGF) in the peritoneum.

31. The method according to claim 20, wherein the peritoneal therapeutic fluid is used for decreasing long term fibrosis.

32. The method according to claim 20, wherein the biocompatibility enhancing agent is pegylated with Polyethyleneglycol (PEG) or Methoxy-Polyethyleneglycol (mPEG), provided that the BCA is not polyethylene glycol (PEG) or a derivative of a polyethylene glycol.

33. The method according to claim 20, wherein the peritoneal therapeutic fluid further comprises at least one ingredient which is selected from the group consisting of alkali metal ions, alkaline earth metal ions, an osmotic agent and a pH-buffer.

34. The method according to claim 20, wherein the peritoneal therapeutic fluid further comprises at least one osmotic agent.

35. The method according to claim 20, wherein the peritoneal therapeutic fluid further comprises glucose.

36. The method according to claim 20, wherein the peritoneal therapeutic fluid further comprises a saccharide selected from the group consisting of fructose, a disaccharide, an oligosaccharide, maltodextrin and a polysaccharide, or any combination thereof.

37. The method according to claim 20, wherein the peritoneal therapeutic fluid is an aqueous solution has an alkali equivalent in an amount of 25 to 50 mEq/L.

38. The method according to claim 20, wherein the electrolytes comprise one or more of sodium, potassium, calcium, and/or magnesium.

39. The method according to claim 21, wherein the peritoneal therapeutic fluid is an aqueous solution comprising: sodium in an amount of 90 to 150 mEq/L; potassium in an amount of 0 to 5 mEq/L; calcium in an amount of 0 to 6 mEq/L; magnesium in an amount of 0 to 4 mEq/L; and an alkali equivalent in an amount of 25 to 50 mEq/L.

40. The method according to claim 21, wherein the peritoneal therapeutic fluid further comprises at least one saccharide.

41. The method according to claim 31, further comprising at least one disaccharide selected from the group consisting of sucrose, Gentiobiulose, Laminaribiose, Gentiobiose, Rutinulose, Xylobiose, trehalose, β,β-Trehalose, α,β-Trehalose, lactulose, sophorose, lactose, cellobiose, chitobiose, maltose, Kojibiose, Nigerose, Isomaltose, Turanose, Maltulose, Palatinose, Mannobiose, Melibiose, Melibiulose, and Rutinose.

42. The method according to claim 21, wherein the peritoneal therapeutic fluid further comprises maltodextrin or at least one oligosaccharide that is a product of limited hydrolysis of one of more of the following: starch, amylose, amylopectin, fructan, glucan, galactan, mannan, cellulose, arabic gum, glycogen, dextran, hemicellulose, arabinoxylose, and pectin, wherein the product of limited hydrolysis has a molecular weight of 90 D to 500 kD.

43. The method according to claim 21, wherein the peritoneal therapeutic fluid further comprises at least one alpha-glucan with a degree of polymerization of 3 or higher and has a molecular weight of 90 D to 500 kD.

44. The method according to claim 21, wherein the peritoneal therapeutic fluid further comprises at least one saccharide which is selected from the group consisting of isomaltotriose, nigerotriose, maltotriose, melezitose, maltotriulose, raffinose, kestose, maltodextrin, dextrins, heparin, Dextran, glycogen, pullulan, starch, amylose, amylopectin, icodextrin, and combinations thereof.

45. The method according to claim 21, wherein the peritoneal therapeutic fluid further comprises at least one saccharide which has a molecular weight in a range of 90D to 50 kD, 90D to 500 D, 90D to 1.5 kD, 1.5 kD to 50 kD, 350D to 50 kD, 250D and 50 KD, or 150 to 400 D.

46. The method according to claim 21, wherein the peritoneal therapeutic fluid further comprises at least one saccharide, and wherein the at least one saccharide is present in a total concentration of ≥20.02% by weight (200 mg/L), ≥0.75% by weight, ≥2.4% by weight, ≥5% by weight, ≥7.5% by weight, or ≥20% by weight.

47. The method according to claim 21, wherein the peritoneal therapeutic fluid is used for decreasing expression of Vascular Endothelial Growth Factor (VEGF) in the peritoneum.

48. The method according to claim 21, wherein the peritoneal therapeutic fluid is used for decreasing long term fibrosis.

49. The method according to claim 21, wherein the biocompatibility enhancing agent is pegylated with Polyethyleneglycol (PEG) or Methoxy-Polyethyleneglycol (mPEG), provided that the BCA is not polyethylene glycol (PEG) or a derivative of a polyethylene glycol.

50. The method according to claim 21, wherein the peritoneal therapeutic fluid further comprises at least one ingredient which is selected from the group consisting of alkali metal ions, alkaline earth metal ions, an osmotic agent and a pH-buffer.

51. The method according to claim 21, wherein the peritoneal therapeutic fluid further comprises at least one osmotic agent.

52. The method according to claim 21, wherein the peritoneal therapeutic fluid further comprises glucose.

53. The method according to claim 21, wherein the peritoneal therapeutic fluid further comprises a saccharide selected from the group consisting of fructose, a disaccharide, an oligosaccharide, maltodextrin and a polysaccharide, or any combination thereof.

54. The method according to claim 21, wherein the peritoneal therapeutic fluid is an aqueous solution has an alkali equivalent in an amount of 25 to 50 mEq/L.

55. The method according to claim 21, wherein the electrolytes comprise one or more of sodium, potassium, calcium, and/or magnesium.

Description

BRIEF DESCRIPTION OF FIGURES

(1) FIG. 1 Comparative testing of PDFs after 48 hours results in decreased resazurin to reorufin conversion.

(2) FIG. 2 Results of resazurin to reorufin conversion, Resveratrol, Polydatin, PEG, PD solution #1.

(3) FIG. 3 Results of resazurin to reorufin conversion, Resveratrol, Polydatin, PEG, PD solution #2.

(4) FIG. 4 Results of resazurin to reorufin conversion, Resveratrol, Polydatin, PEG, PD solution #3.

(5) FIG. 5 Results with Medium control.

(6) FIG. 6 Results of resazurin to reorufin conversion, Resveratrol in different PD solutions.

(7) FIG. 7 Results of resazurin to reorufin conversion, Piceatannol in different PD solutions.

(8) FIG. 8 Results of resazurin to reorufin conversion, Pterostilbene in different PD solutions.

(9) FIG. 9A, 9B Results of resazurin to reorufin conversion, Piceid in different PD solutions.

(10) FIG. 10 Results of resazurin to reorufin conversion, Caffeic acid in different PD solutions.

(11) FIG. 11 Results of resazurin to reorufin conversion, Luteolin in different PD solutions.

(12) FIG. 12 Results of resazurin to reorufin conversion, Delphinidin in different PD solutions.

(13) FIG. 13 Results of peritoneal VEGF expression in Sprague-Dawley rats after 2 to 4 weeks Peritoneal Dialysis with PD solution #4 in absence or presence of Resveratrol 40 μM (average concentrations and standard deviations).

(14) The following Examples illustrate embodiments of the present invention:

EXAMPLES

(15) Molecular Weight Measurement:

(16) The saccharides are dissolved in extra-pure water in a concentration of 0.5% (w/v). The solutions are heated at 95° C. for 30 minutes. The polymers are analyzed using the following devices: Alliance chromatography system (Waters corporation, Milford, Mass., USA), DAWN-EOS light scattering detector (Wyatt Technology, Santa Barbara, USA) with λ0=658 nm and 16 detectors in the range of angles from 14.4 to 163.3°, K5 flow cell. The polymers are fractionated on a precolumn and three columns having the separation ranges 300-10.sup.4, 5×10.sup.4-2×10.sup.6 and 10.sup.6-10.sup.8 (SUPREMA-Gel, PSS Polymer Standards Service GmbH, Mainz, Germany). 100 μl of solution are injected. The fractionation takes place at a temperature of 30° C. and a flow rate of 0.8 ml/min with 0.05M NaNO3 as eluent. The Astra V 5.1.8.0 program (from Wyatt Technology, Santa Barbara, USA) is used to analyze the molecular weight distribution of the samples. Same procedure can be used when molecular weight of other compounds than saccharides are measured.

(17) Dialysis Solutions:

(18) In accordance with this invention, peritoneal dialysis fluids are provided, containing an osmolality sufficient to cause diffusion of water and waste products across the peritoneum after infusion of the peritoneal dialysis fluid into the peritoneal cavity of a patient. In addition to an osmotic agent or a combination of osmotic agents, the present peritoneal dialysis fluid contains amounts of various physiologically important electrolytes in concentrations comparable to those in plasma. A suitable peritoneal dialysis fluid has been described in the definitions part of this patent.

(19) TABLE-US-00001 TABLE I PD Sol PD#1 PD#2 PD#3 PD#4 Osmolality StaySafe ® Physioneal ® Extraneal ® StaySafe ® (mOsm/kg) 346 485 284 486 Osmotic Glu Glu Ico Glu Agent (%) w/v 1.25 3.86 7.5 4.25 Sodium 132 132 133 132 (mEq/L) Calcium 3.5 1.75 3.5 3.5 (mEq/L) Magnesium 0.5 0.25 0.5 0.5 Chloride 96 101 96 96 Lactate 40 10 40 40 Bicarbonate 25 pH 5.5 pH 7 ph 5.5 pH 5.5 tested BCA /, R, P, PE /, R, P, PE /, R, P, PE /, P, Pa, Pt, CA, Pa, Pt, CA, Lu, De Lu, De Legend to Table I: Solutions tested for their application as peritoneal dialysis fluids. Abbreviations: Glu, glucose; Ico, icodextrin; OsAg, osmotic agent; BCA, added “biocompatibility enhancing agent”. Concentrations in % (w/v) and mEq/L; osmolality in mOsm/kg. Tested BCAs are: The stilbenoids Resveratrol (R), Piceid (Polydatin) (P), Piceatannol (Pa), Pterostilbene (Pt); the phenolic acid Cafeic Acid (CA), the flavonoides Luteolin (Lu), Quercetin (Qu), Delphinidin (De). PEG 1450 Carbowax (PE). Legend to Table I: Solutions tested for their application as peritoneal dialysis fluids. Abbreviations: Glu, glucose; Ico, icodextrin; OsAg, osmotic agent; BCA, added “biocompatibility enhancing agent”. Concentrations in % (w/v) and mEq/L; osmolality in mOsm/kg. Tested BCAs are: R Resveratrol, P Piceidand PE PEG 1450 Carbowax.

(20) Table 1 shows peritoneal dialysis fluids, compared for testing the effect of reduction of cytotoxicity by addition of tested BCAs. The study involves evaluation of additions of BCAs at different concentrations to PD solutions.

(21) StaySafe 1.25 solution was chosen to show impact of acidic pH at low Glucose concentration in an environment of high lactate buffer. Physioneal 3.86 was chosen to show the impact of high glucose concentration at physiological pH in an environment of low lactate buffer. StaySafe 4.25 was chosen to show combined challenge of acidic pH and high glucose concentration. Extraneal was chosen to compare the difference of glucose and maltodextrin at acidic pH and at high lactate concentration.

(22) The examples show that addition of specifically selected BCAs increase biocompatibility of currently marketed PDFs. Those skilled in the art readily understand that addition of such “biocompatibility enhancing agents” will increase long term biocompatibility of any peritoneal therapeutic and/or dialysis solution, more specifically of such solutions containing sugar and/or sugar polymer-derived osmotic agents or such, and this even in cases and models where certain dialysis solutions do not show immediate cytotoxicity and/or very low AGE formation.

(23) Solutions are applied to different toxicity experiments in absence or presence of specifically selected BCAs, to show that BCAs, exemplifying the present invention, decrease cytotoxic side-effects, and thereby increasing biocompatibility, as compared to reference solutions without such BCAs.

(24) Toxicity:

(25) The following experiments compare the cytotoxicity of reference solutions in absence or presence of BCAs of this invention, to show increased biocompatibility of dialysis solutions in presence of BCAs of this invention.

Examples 1, 2, 3, and 4

(26) Experimental comparison of different dialysis solution with respect to their effect on human peritoneal mesothelial cells, applying the following protocol.

(27) Cell Culture

(28) Experimental Procedure:

(29) Human peritoneal mesothelial cells (HPMC) were purchased from Zen Bio Inc. and cultured in cell culture flasks using suppliers media. Near confluent HPMC were harvested by trypsinization, seeded into collagen-coated 96-well tissue culture plates (Corning) and allowed to adhere overnight. The medium was changed to twice diluted with dialysis solution for 48 up to 72 hours.

(30) Cell viability was established applying the promega resazurin assay, following the suppliers protocol. Living cells are metabolically active and are able to reduce the non-fluorescent dye resazurin to the strongly-fluorescent dye resorufin. The fluorescence output is proportional to the number of viable cells over a wide concentration range. This also allows the calculation of the proliferation rate for cells capable of consecutive cell division. Resazurin is effectively reduced in mitochondria making it also useful to assess mitochondrial metabolic activity. For the dose-response relationship, relative viability was plotted against the test item concentrations.

(31) In the case of Piceid, the intra-cellular ATP level was determined with the CTG assay. For this, media was completely removed from all wells by aspiration, 60 μl of CTG reagent was added to each well, and incubated for 5 min at RT while softly shaking (50 rpm). Using a Victor3 1420 Multilabel Counter, the emitted luminescence produced in the CTG assay was measured. For the dose-response relationship, absolute luminescence (background subtracted) was related to the negative (medium) control and relative viability values were plotted against the test item concentrations. For the dose-response relationship, absolute luminescence (background subtracted) was related to the negative (medium) control and relative viability values in presence of BCA were plotted against the BCA concentrations.

(32) All assays were conducted in a duplex format using the same cell culture.

(33) Results:

Example 1

(34) Comparative testing of PDFs after 48 hours results in decreased resazurin to reorufin conversion, which translates to decreased cell-viability. See FIG. 1.

Example 2

(35) Addition of selected BCAs of this invention partially reestablished reszurin to reorufin conversion, which is interpreted as a result of a decreased cytotoxicity, due to the application of the tested BCAs. Compounds were added at 9 dilutions (Cmax=500 μM) together with tested PD solutions or Medium control. Incubation was 48 hours.

(36) Results with PD-Solution #1 are presented in FIG. 2

(37) Resveratrol improves cell viability of HPMC cells up to 20%. Piceid (polydatin) shows minor improvements.

(38) Results with PD-Solution #2 are presented in FIG. 3.

(39) Resveratrol improves cell viability of HPMC cells up to 40%. Piceid (polydatin) shows minor improvements.

(40) Results with PD-Solution #3 are presented in FIG. 4.

(41) Resveratrol improves cell viability of HPMC cells up to 40%. PEG shows minor improvements.

(42) Medium Control is presented in FIG. 5:

(43) In control medium, without cytotoxic stress, resveratrol, piceid (polydatin) and PEG have no significant effect on cell viability until Cmax.

(44) In conclusion, we obtained a strong effect of Resveratrol reducing cyto-toxicity of all three tested peritoneal dialysis solutions and a minor effect of piceid. A possible explanation for a relatively weaker effect of piceid is, that piceid has first to be converted to resveratrol or another biological active compound by enzymes that are present in the peritoneum. We therefore shall show a stronger effect of piceid in an animal model.

(45) For PD-solution #3 we observed a cytotoxicity decreasing effect of PEG. We had used PEG simply as a control in our experiments and have no explanation for this observation.

Example 3

(46) Addition of selected BCA resveratrol partially reestablished resazurin to reorufin conversion, in a triplicate assay, which is interpreted as a result of a decreased cytotoxicity, due to the application of the tested BCA. In this series, Resveratrol was added 5 minutes in advance to application of test-solutions, at 9 dilutions (Cmax=500 μM). Incubation was 72 hours. Results are presented in FIG. 6.

(47) Resveratrol improves viability of HPMC cells exposed to PD-Solution #1 by up to 84%.

(48) Resveratrol improves viability of HPMC sells exposed to PD-Solution #2 by up to 28%.

(49) Resveratrol improves viability of HPMC cells exposed to PD-Solution #3 by up to 105%.

Example 4

(50) Addition of selected BCAs, namely of

(51) the stilbenoids Piceatannol (Pa), Pterostilbene (Pt), Piceid (Polydatin) (P);

(52) the phenolic acid Cafeic Acid (CA);

(53) the flavonoides Luteolin (Lu), Delphinidin (De);

(54) partially reestablished resazurin to reorufin conversion, or partly re-established intracellular ATP-level, which is interpreted as a result of a decreased cytotoxicity, due to the application of the tested BCA. Test items were tested at 3 replicates per concentration. All assays were conducted in a duplex format using the same cell culture. Incubation was 72 hours.

(55) Results with Piceatannol are presented in FIG. 7

(56) Piceatanol improves cell viability of HPMC cells, when exposed to PD-Solution #3 by up to 44%, and when exposed to PD-Solution #4 by up to 40%.

(57) Results with Pterostilbene are presented in FIG. 8

(58) Pterostilbene improves cell viability of HPMC cells, when exposed to PD-Solution #3 by 183%, and when exposed to PD-Solution #4 by 118%.

(59) Results with Piceid (Polydatin) are presented in FIGS. 9a. and b. In this experimental series, Piceid improved viability of HPMC cells measured by resazurin to resorufin transformation, when exposed to Solution #3 by up to 32%, when exposed to PD-Solution #4 by up to 17% (FIG. 9a). Measured by ATP-level re-establishment, Piceid improves viability of HPMC cells exposed to PD-Solution #4 by 51%.

(60) Results with Cafeic Acid are presented in FIG. 10.

(61) Cafeic Acid improves cell viability of HPMC cells, when exposed to PD-Solution #3, up to 32%. Cell viability improvement is minor when HPMC cells are exposed to PD-Solution #4.

(62) Results with Luteolin are presented in FIG. 11.

(63) Luteolin improves cell viability of HPMC cells, when exposed to PD-Solution #3 by up to 56%, and when exposed to PD-Solution #4 by up to 21%.

(64) Results with Delphinidin are represented in FIG. 12.

(65) Delphinidin improves cell viability of HPMC cells, when exposed to PD-Solution #3 by up to 57%. No cell viability improvement du to Delphinidin was observed under the applied experimental conditions, when testing HPMC cells expose to PD-Solution #4.

(66) Taken together, results from examples 1 to 4 indicate a general effect of tested BCAs by increasing cell-viability of HPMC cells, when exposed to PD-Solutions. For most BCAs the concentration of maximal activity varies between 0.08 μM and 18.5 μM, but in some cases concentrations of 167 or even 500 μM were highly efficacious. For those skilled in the art such variability of concentration with highest efficacy is not surprising, reflecting different bioavailbilities and target affinities. Nevertheless, such a general impact of so many representatives of given classes of naturally occurring compounds within the same model is a striking discovery.

(67) All tested compounds (Polyphenols) showed some improvement of HPMCs when exposed to at least one of the 4 tested PD-Solutions. All tested Stilbenoids (Resveratrol, Piceid, Piceatanol and Pterostlben) increased cell viability as well on Glucose based as ond Icodextrin based PD-Solutions.

(68) The phenolic acid Cafeic Acid, and flavanoides Luteolin and Delphinidin mainly improved Icodextrin based dialysis solutions.

(69) Those, skilled in the art understand that a toxicity cell model is a relatively fragile model, and that measurable cell-culture toxicity decrease is already dependent on measurable cell-toxicity in the first place. Nevertheless we observed overall higher stress due to Icodextrin based PTFs as compared to Glucose Based PTFs, under the applied experimental conditions. Such stronger toxicity challenge enabled us to show BCA activity of tested compounds over a larger range of concentrations. The results of Piceid show highest variation of all tested compounds. We believe that the need of metabolization of piceid, dependent on metabolic capacity of cultured cells, might be a reason for such variability. In example 4 we succeeded to show reproducible BCA activity of Piceid in 3 different experimental set-ups.

Example 5

(70) Animal Studies have been carried out as described in Lee et al. 2012:

(71) Experimental Procedure:

(72) Peritoneal access ports were inserted in male Sprague-Dawley rats. After one week, rats started to received peritoneal treatment: 10 rats receive once daily 20 ml of Sol #4, 10 rats received 20 ml of Sol #4 with addition of selected BCA (resveratrol), during 2 hour infusions. After 2 to 4 weeks, the abdomen was opened, the peritoneum was recovered and submitted to protein extraction. Tissue VEGF concentration was established by ELISA (Abcam Rat VEGF ELISA Kit, ab100787) on obtained protein preparations (pg/ml).

(73) Results:

(74) Increased VEGF expression after chronic peritoneal dialysis has been reported in humans and rat-models, and is related to fibrosis and angiogenesis as side effects of long term peritoneal dialysis treatment (Zweers, 2001; Park, 2004). Results of example 5 (table II and FIG. 13) show that addition of selected BCA (resveratrol) decreases expression of VEGF in the peritoneum of standard PDF treated rats, indicating improved biocompatibility of BCA supplemented PDFs in the animal model.

(75) TABLE-US-00002 TABLE II VEGF expression in peritoneal tissue after 2 or 4 weeks of peritoneal dialysis wit Solution #4 in absence or presence of Resveratrol 40 μM. Values between 2 and 4 weeks were highly reproducible and therefore combined for the statistical analysis. VEGF concentration Treatment (pg/mL) in Statistical PD Solution (weeks) Animal prot. prep. results Sol #4 2 1 81.64 Average 2 80.21 95.18 3 93.52 Stand. Dev. 4 92.56 20.08 5 84.42 4 6 84.42 7 136.91 8 95.72 9 77.05 10 125.32 Sol #4 + 40 μM 2 11 50.21 Average Resveratrol 12 39.99 60.94 13 70.88 Stand. Dev. 14 77.84 10.65 15 60.12 4 16 62.02 17 63.71 18 57.91 19 68.45 20 58.23 t-test pval 0.00065

REFERENCES

(76) Barre, Chen, Cooker, Moberly. Adv Perit Dial. 1999; 15:12-6. Catalan, Reyero, Egido, Ortiz. J Am Soc Nephrol. 2001; 12(11):2442-9. Ha, Yu, Choi, Cha, Kang, Kim, Lee. Perit Dial Int. 2000; 20 Suppl 5:S10-18. Konings, Schalkwijk, van der Sande, Leunissen, Kooman. Perit Dial Int. 2005; 25(6):591-5. Lee, Kang, Kim, Nam, Paeng, Kim, Li, Park, Kim, Han, Yoo, Kang, 2012, Laboratory Investigation 92: 1698-1711. Mangram, Archibald, Hupert, Tokars, Silver, Brennan, Arduino, Peterson, Parks, Raymond, McCullough, Jones, Wasserstein, Kobrin, Jarvis. Kidney International, Vol. 54 (1998), pp. 1367-1371 Park, Lee, Kim, Kim, Cho, Kim. Perit Dial Int. 2004; 24(2):115-22. Moriishi, Kawanishi. Perit Dial Int. 2008; 28 Suppl 3:S96-S100. ter Wee, Ittersum. Nat Clin Pract Nephrol. 2007; 3(11):604-12. Williams, Craig, Topley, Von Ruhland, Fallen, Newman, Mackenzie, Williams. J Am Soc Nephrol 2002; 13:470-479. Zweers, Struijk, Smit, Krediet. 2001. J Lab Clin Med. 137(2):125-32.