Composition for maintaining organ and cell viability
The present invention is directed to nanoparticle compositions for maintaining organ, tissue and cellular viability when such are separated from normal physiological supports. Compositions containing the nanoparticle compositions and methods of preserving organs such as kidneys, both in vivo and ex vivo, are also disclosed.
1. A two-phase composition comprising:
a first phase comprising a base nutritive medium; and
a second phase comprising nanoparticles that comprise an outer lipophilic coating and an inner hydrophilic core, wherein
a) the first phase comprises physiologically compatible concentrations of water soluble or dispersible nutrients, and physiological salts;
b) the nanoparticles of the second phase comprise lipids, fatty acids, sterols and free fatty acids; and
c) the two-phase composition has an osmolality of at least about 300 mOsM/kg, and
wherein the nanoparticles have a mean diameter ranging from about 100 nm to about 300 nm, and wherein the hydrophilic inner core of the nanoparticle contains a solution or suspension comprising a moiety capable of binding and releasing oxygen, and the first phase comprises a composition comprising Table 2A or Table 6, wherein
Table 2A is:
g/Liter
RANGE
Adenosine
0.950–1.050
Adenosine 5′ Monophosphate
0.0019–0.0021
Adenosine Triphosphate
0.0019–0.0021
Allopurinol
0.133–0.147
B′ Nicotinamide Adenine
0.038–0.042
Dinucleotide Phosphate
B′Nicotinamide Adenine
0.0019–0.0021
Dinucleotide
Calcium Chloride
0.152–0.168
Choline Chloride
0.0085–0.0094
Chondrotin Sulfate
0.0038–0.0042
Cocarboxylase
0.038–0.042
Coenzyme A
0.0095–0.00105
Cyclodextrin
0.475–0.525
Deoxyadenosine
0.038–0.042
Deoxycytidine
0.038–0.042
Deoxyguanosine
0.038–0.042
Dextran 70
33.25–36.75
Flavin Adenine Dinucleotide
0.038–0.042
Folic Acid
0.0026–0.0028
Glucose
3.800–4.200
Glutathione
0.950–1.050
Glycine
0.0179–0.0197
Table 6 is:
g/Liter
Soy Hydrolysate
6.0–10.0
Glutathione Reduced
0.008–0.020
Vitamin A Acetate
0.0002–0.0003
Vitamin C
0.040–0.060
Vitamin E
0.00002–0.00003
Catalase
0.040–0.060
SOD
0.040–0.060
L-Cysteine HCl
0.040–0.060
Taurine
0.025–0.040
Methionine
0.015–0.030
Zinc Sulfate
0.0008–0.0009
Selenium
0.000025–0.000035
Cupric Sulfate
0.0000045–0.000006
Ethanolamine
0.0025–0.005
Mercaptoethanol
0.003–0.006.
2. The two-phase composition of claim 1 wherein the two-phase composition has an osmolality ranging from about 385-425 mOsM/kg.
3. The two-phase composition of claim 1 wherein the pH of thereof is from about 7.2 to about 7.4.
4. The two-phase composition of claim 1 wherein the nanoparticles have a mean diameter ranging from about 100 nm to about 300 nm.
5. The two-phase composition of claim 4 wherein the nanoparticles have a mean diameter ranging from about 100 nm to about 200 nm.
6. The two-phase composition of claim 1 , further comprising cellular growth factors.
7. The two-phase composition of claim 6 , wherein said cellular growth factors are selected from the group consisting of epithelial and endothelial growth factors, vascular endothelial growth factors, platelet derived endothelial growth factors, epithelial growth factors, hepatocyte growth factors, and mixtures thereof.
8. The two-phase composition of claim 1 which comprises about equal amounts of the first phase and the second phase.
9. The two-phase composition of claim 1 wherein the outer lipophilic coating comprising a free fatty acid selected grom the group consisting of oleic acid and linoleic acid, and combinations thereof.
10. The two-phase composition of claim 1 , which, when comprising Table 2A, further comprises Table 1
g/Liter
RANGE
Adenine HCl
0.00019–0.00021
vitamin B-12
0.00065–0.0007
Biotin
0.00000038–0.00000042
Cupric Sulfate
0.00000124–0.00000137
Ferric Nitrate
0.000048–0.000053
Ferric Sulfate
0.00048–0.00053
Putrescine HCl
0.000077–0.000085
Pyridoxine HCl
0.000029–0.000033
Riboflavin
0.00021–0.000231
Thymidine
0.00035–0.00039
Zinc Sulfate
0.00041–0.000454.
11. The two-phase composition of claim 1 , which when comprising Table 2A further comprises Table 2B,
wherein Table 2B is
g/Liter
RANGE
Heparin
0.171–0.189
HEPES
3.396–3.753
Hypoxanthine
0.002–0.0022
Inositol
0.0124–0.0137
Insulin
0.0095–0.0105
L-Alanine
0.00428–0.00473
L-Arginine
0.141–0.155
L-Asparagine
0.0076–0.0084
L-Aspartic Acid
0.064–0.070
L-Cysteine
0.0297–0.0329
L-Cystine
0.0167–0.0185
L-Glutamic Acid
0.007–0.0078
L-Glutamine
4.750–5.250
L-Histidine
0.030–0.033
L-Isoleucine
0.052–0.0572
L-Leucine
0.057–0.063
L-Lysine
0.0095–0.0105
L-Methionine
0.019–0.021
L-Phenylalanine
0.0337–0.0373
L-Proline
0.0164–0.0182
L-Serine
0.025–0.0276
L-Threonine
0.051–0.056.
12. The two-phase composition of claim 1 , which when comprising Table 2A further comprises Table 2C,
wherein Table 2C is
g/Liter
RANGE
L-Tryptophan
0.009–0.0095
L-Tyrosine
0.053–0.059
L-Valine
0.050–0.055
Magnesium Chloride
0.058–0.0643
Magnesium Sulfate
0.0475–0.0525
Mannose
3.135–3.465
Niacinamide
0.0019–0.0021
Pantothenic Acid
0.0021–0.0024
Potassium Chloride
0.296–0.328
Pyridoxal HCl
0.0019–0.0021
Pyruvic Acid
0.209–0.231
Sodium Bicarbonate
1.140–1.260
Sodium Chloride
6.650–7.350
Sodium Phosphate Dibasic
0.0676–0.0748
Sodium Phosphate Monobasic
0.0516–0.0570
Thiamine
0.0021–0.0023
Transferrin
0.00475–0.00525
Uridine
0.038–0.042
Uridine Triphosphate
0.038–0.042
Yeastolate Ultra-Filtered
38–42 ML.
13. The two-phase composition of claim 1 , which when comprising Table 2A further comprises Table 3,
wherein Table 3 is
g/Liter
RANGE
L-Cystine
0.0167–0.0185
L-Tyrosine
0.053–0.059.
14. The two-phase composition of claim 1 , wherein the second phase further comprises Table 4 when the first phase comprises Table 2A, or further comprises Table 9 when the first phase comprises Table 6, and wherein
Table 4 is
g/Liter
RANGE
Cholesterol
0.00475–0.00525
Cod Liver Oil
0.00095–0.00105
Epithelial Growth Factor
0.00000285–0.00000315
Hepatocyte Growth Factor
0.0000048–0.0000053
Hydrocortisone
0.00095–0.00105
Linoleic Acid
0.00095–0.00105
Linolenic Acid
0.00095–0.00105
Oleic Acid
0.00095–0.00105
Phosphatidylcholine
0.6%
Platelet Derived
0.00000095–0.00000105
Endothelial Growth Factor
Pluronic F-68
0.950–1.050
Prostaglandin E1
0.000042–0.0000263
Triiodo-L-Thyroxine
0.00000475–0.0000053
TWEEN 80 ®
0.002375–0.002625
Vascular Endothelial Growth
0.0000046–0.00000525
Factor
Vitamin E
0.0019–0.0021
wherein Table 9 is
ETOH 95%
8.00–16.00 ml
Soy Hydrolysate
3–8 ml
Phosphatidyl Choline
0.00095–0.010 g/liter
Arachadonic Acid
0.0000015–0.00002 g/liter
Linoleic Acid
0.000095–0.00015 g/liter
Linolenic Acid
0.000095–0.00015 g/liter
Myristic Acid
0.000095–0.00015 g/liter
Oleic Acid
0.000095–0.00015 g/liter
Palmitic Acid
0.000095–0.00015 g/liter
Stearic Acid
0.000095–0.00015 g/liter
Cholesterol
0.002–0.004 g/liter
Vitamin E
0.0006–0.0008 g/liter
Tween 80
0.020–0.030 g/liter
Pluronic F-68
0.010–1.000 g/liter.
15. The two-phase composition of claim 1 , which, when comprising Table 6, further comprises Table 7A,
wherein Table 7A is
g/Liter
RANGE
Sodium Gluconate
18.500–25.000
Potassium Phosphate
3.000–4.500
Magnesium Sulfate
1.000–1.500
Calcium Chloride
0.100–0.150
Sodium Phosphate Monobasic
0.250–0.350.
16. The two-phase composition of claim 15 , which further comprises Table 7B,
wherein Table 7B is
g/Liter
RANGE
L-Arginine HCl
0.065–0.080
L-Aspartic Acid
0.050–0.065
L-Glutamic Acid
0.100–0.160
L-Glutamine
0.300–0.400
Glycine
0.045–0.060
L-Histidine HCl—H 2 O
0.155–0.170
L-Isoleucine
0.025–0.030
L-Leucine
0.045–0.055
L-Lysine HCl
0.240–0.300
L-Phenylalanine
0.045–0.055
L-Proline
0.045–0.055
L-Threonine
0.065–0.075
L-Tryptophan
0.035–0.0450
L-Valine
0.055–0.070
L-Cystine
0.018–0.024
L-Tyrosine
0.050–0.060.
17. The two-phase composition of claim 15 , which further comprises Table 7C
wherein Table 7C is
g/Liter
RANGE
Glucose
4.500–6.000
Mannose
8.000–12.000.
18. The two-phase composition of claim 15 , which further comprises Table 8, wherein
Table 8 is
g/Liter
RANGE
Biotin
0.000015–0.00003
Choline Bitartrate
0.40–0.50
Folic Acid
0.00035–0.0005
Inositol
0.0008–0.002
Niacinamide
0.0008–0.002
Pantothenic Acid
0.0008–0.002
Pyridoxine
0.0008–0.002
Riboflavin
0.0008–0.002
Thiamine
0.008–0.015
Vitamin B-12
0.000015–0.0003
Adenosine
0.85–1.50.
19. The two phase composition of claim 1 wherein the moiety capable of binding and releasing oxygen is a heme protein.
20. The two phase composition of claim 1 wherein said hydrophilic inner core contains a biologically active moiety.
21. The two-phase composition of claim 20 wherein said biologically active moiety is selected from the group consisting of chemotherapeutic agents, proteins, peptides, polypeptides, enzymes and mixtures thereof.
22. The three phase composition comprising the composition of claim 1 admixed with a composition comprising a nonoparticle having an outer lipophilic coating and a hydrophilic inner core comprising a solution or suspension comprising a moieity capable of binding and releasing oxygen.
23. The three phase composition of claim 22 wherein the moiety capable of binding and releasing oxygen is a heme protein.
24. A three phase composition comprising the composition of claim 1 admixed with a composition comprising a nanoparticle having an outer lipophilic coating and a hydrophilic inner core comprising a biologically active moiety.
25. A process for preparing the two phase composition of claim 1 comprising combining a liquid first phase comprising a base nutritive medium containing physiologically compatible concentrations of water soluble or dispersible nutrients, and physiological salts with a liquid second phase comprising nanoparticles that comprise an outer lipophilic coating and an inner hydrophilic core under conditions sufficient to prepare a final two phase composition having an osmolality of at least about 300 mOsM/kg.
26. A method of preserving a mammalian cells or tissues, comprising placing said cells or said tissue in a sufficient amount of the composition of claim 1 .
27. A method of preserving a mammalian kidney, ex vivo, comprising perfusing the kidney with an effective amount of the composition of claim 1 .
28. A method of maintaining the health of a member of the group consisting of mammalian cells and tissues ex vivo, comprising placing said member in a sufficient amount of a composition of claim 1 and maintaining said member at a non-hypothermic temperature.
29. The method of claim 28 wherein non-hypothermic temperature ranges from about 20 to 37° C.