Bear derived isolate and method
First Claim
1. A composition of matter comprising pharmacological properties and which is a deproteinated isolate which has been obtained from a sample of urine or serum taken from a fasting bear from which food has been withheld for two weeks or more, which sample has been subjected to deproteination, then the deproteinated isolate having the pharmacological properties of inducing, when injected into another mammal, conditions observable in denning black bears including reduced heart rate, temperature reduction, or a tranquility distinguishable from normal behavior.
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Abstract
A bear derived isolate found in denning black bears or black bears from which food has been withheld for at least two weeks when injected into another mammal produces reduced heart rate, reduced respiratory rate, reduced temperature, and a wakeful tranquility not unlike the phenomena of the denning black bear. The isolate is taken from either the serum or urine of such denning bear. The method of the invention involves the steps of fractionating the serum or urine of a denning black bear to produce fractions of varying potency.
11 Citations
22 Claims
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1. A composition of matter comprising pharmacological properties and which is a deproteinated isolate which has been obtained from a sample of urine or serum taken from a fasting bear from which food has been withheld for two weeks or more, which sample has been subjected to deproteination, then the deproteinated isolate having the pharmacological properties of inducing, when injected into another mammal, conditions observable in denning black bears including reduced heart rate, temperature reduction, or a tranquility distinguishable from normal behavior.
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2. A method for obtaining an isolate form the blood or urine of a fasting bear which bear has not eaten for two weeks or more, such isolate being sufficiently free of impurities for repeated administration to mammals to induce activity of a kind observed in denning bears comprising the steps of:
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drawing a sample of blood or urine from said bear, deproteinating and extracting the isolate from such sample with organic solvents, further purifying the presence of said isolate by countercurrent chromatography, flash column chromatography, preparative thin layer chromatography, and/or high performance liquid chromatography, and testing the purity of the isolate so obtained by TLC and/or chemical or spectroscopic detection.
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3. A deproteinated urine or serum of a fasting bear which has not eaten for two weeks or more comprising a therapeutic compound producing, in any combination, the following behavior in another mammal:
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tranquility, or reduced heart rate, or increased osteoblastic activity, or decreased osteoclastic activity.
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4. A composition of matter having the characteristics of deproteinated urine or serum of a fasting bear, which bear has not eaten for two weeks or more and capable of producing the following behavior in a guinea pig injected with said composition produces the following:
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tranquility, or reduced heart rate, or increased osteoblastic activity, or decreased osteoclastic activity.
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5. A composition of matter having the characteristics of deproteinated urine or serum of a fasting bear, which bear has not eaten for two weeks or more and capable of producing when injected in a guinea pig:
enhanced bone remodeling.
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6. A composition of matter having the characteristics of deproteinated urine or serum of a fasting bear, which bear has not eaten for two weeks or more and capable of producing when injected in an ovariectomized rat:
enhanced bone formation.
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7. A composition of matter comprising the deproteinated urine or serum of a denning black bear, which denning black bear neither eats, drinks, urinates, or defecates for lengthy periods of time having the following properties:
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soluble in water, methanol, and 1-butanol, insoluble in less polar organic solvents including ethyl acetate, chloroform, toluene, and hexane, stable at room temperature for four days or more, heat resistant to 65°
C., andstable when frozen in a light resistant container under nitrogen gas which, when injected into a guinea pig, is capable of producing reduced heart rate, reduced temperature, or observable tranquility differing from normal. - View Dependent Claims (8, 9)
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10. A pharmacological composition of matter comprising the capability of enhancing bone formation in ovariectomized rats taken from a substance present in the blood or urine of fasting bears, which when fasting are unique in that they have not eaten for two weeks or more, said composition including a quantity of resorptive form of 24,25-dihydroxyvitamin D3 which stimulates bone formation.
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11. The method of producing a pharmaceutical composition from the blood or urine of a fasting bear, which bear has not eaten for two weeks or more, comprising the steps of:
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harvesting the blood or urine from said bear, using counter current chromatography (CCC) to divide the thus withdrawn composition from the bear into 10 factions; and
isolating the inhibitors of bone formulation in Fractions I, II, and III, and purifying the Fractions V, VI, and VII that contain potent stimulation of bone formation, both in the stimulation and proliferation of osteoblast and fibroblasts as well as containing inhibitors to osteoclastic formation and direct inhibitors of resorption by osteoclast.
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12. A composition of matter having the characteristics of deproteinated urine of a fasting bear, which bear has not eaten for two weeks or more and capable of producing the following phenomena in a guinea pig when injected with said composition produces the following:
a decrease in the urea creatinine ratio to a level of 10;
1 or less.
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13. A composition of matter having the characteristics of deproteinated urine of a fasting bear, which bear has not eaten for two weeks or more, and which bear during the period of two weeks or more has reduced its urea to creatinine ratio by at least 25%, capable of producing the following phenomena in a guinea pig when injected with said composition:
a reduction in the urea creatinine ratio to a level of 10;
1 or less.
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14. A composition of matter having the characteristics of deproteinated urine of a fasting bear, which bear has not eaten for two weeks or more, and which produces the following phenomena when injected into a guinea pig:
a reduction in the urea creatinine ratio of said Guinea pig from the pre-injection urea creatinine ratio of the subject Guinea pig.
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15. A composition of matter having the characteristics of unprocessed urine of fasting bears, which bears have been fasting for at least two weeks, but without withholding water and when subjected to alkaline phosphatase as a means of evaluating osteoblastic activity, exhibits and increase in osteoblastic activity of at least 50% above a control.
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16. A composition of matter having the characteristics of a bear derived isolate (BDI) which is isolated from fasting black bears which decreases osteoclastic activity while simultaneously stimulating osteoblastic activity thereby promoting enhanced bone remodeling.
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17. A method of preparing a bear derived isolate (BDI) from the urine of a bear which either dens without eating, drinking or defecating for weeks on end or when the bear has been fasting for at least two weeks but water is not withheld which, when subjected to tests which are in vivo studies with guinea pigs, in vivo studies with rats, in vitro organ studies of calaverial mass bone, and in vitro studies of prevention and proliferation of cells that resorb bone and stimulation of proliferation of cells that form bones using cell cultures of monocytes, osteoclasts, osteoblasts, and fibroblast and when tested by the various said tests produce the following phenomena:
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reduced heart rate;
reduced temperature;
reduced blood pressure;
reduced osteoclastic activity;
increased osteoblastic activity; and
a decrease in the urea/creatinine ratio.
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18. A composition of matter having the characteristics of deproteinated urine or serum of a fasting bear, which bear has not eaten for two weeks or more and capable of producing the following behavior in a guinea pig injected with said composition produces the following:
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tranquility, or reduced heart rate, or increased osteoblastic activity, or decreased osteoclastic activity, or produces at least one of the phenomena as exhibited by a denning black bear.
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19. A method of preparing a bear derived isolate (BDI) from the urine of a bear which either dens without eating, drinking or defecating for weeks on end or when the bear has been fasting for at least two weeks but water is not withheld which, when subjected to tests which are in vivo studies with guinea pigs, in vivo studies with rats, in vitro organ studies of calaverial mass bone, and in vitro studies of prevention and proliferation of cells that resorb bone and stimulation of proliferation of cells that form bones using cell cultures of monocytes, osteoclasts, osteoblasts, and fibroblast and when tested by the various said tests produce the following phenomena:
increased fibroblastic activity.
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20. A composition of matter having the characteristics of deproteinated urine or serum of a fasting bear, which bear has not eaten for two weeks or more and capable of producing the following behavior in another mammal, conditions observable in denning black bears including:
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reduced heart rate, reduced temperature, tranquility distinguishable from normal behavior.
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21. A composition of matter having the characteristics of deproteinated urine or serum of a fasting bear, which bear has not eaten for two weeks or more and capable of producing the following behavior in another mammal, injected with said composition produces the following induces behavioral characteristics of a denning black bear.
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22. A composition of matter having the characteristics of a deproteinated urine or serum sample of a fasting bear, which bear has not eaten for two weeks or more treated as follows:
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first deproteinating the sample, second separating the sample chromatographically into fractions, and third testing the fractions for a purity of isolation which permits the isolate when administered to a mammal other than a bear to induce behavioral characteristics of a denning bear.
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Specification