U.S. patent application number 14/793657 was filed with the patent office on 2016-12-08 for trivalent chromium compounds, compositions and methods of use.
The applicant listed for this patent is INTERHEALTH NUTRACEUTICALS, INC.. Invention is credited to Debasis Bagchi, Xiaoming X. Chien.
Application Number | 20160354353 14/793657 |
Document ID | / |
Family ID | 38521660 |
Filed Date | 2016-12-08 |
United States Patent
Application |
20160354353 |
Kind Code |
A9 |
Chien; Xiaoming X. ; et
al. |
December 8, 2016 |
TRIVALENT CHROMIUM COMPOUNDS, COMPOSITIONS AND METHODS OF USE
Abstract
The present invention provides chromium compounds, specifically
chromium dinicocysteinate, which possess the ability to improve
insulin sensitization, glucose tolerance, bioavailability,
efficacy, and safety, and methods for their use.
Inventors: |
Chien; Xiaoming X.; (San
Ramon, CA) ; Bagchi; Debasis; (Concord, CA) |
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Applicant: |
Name |
City |
State |
Country |
Type |
INTERHEALTH NUTRACEUTICALS, INC. |
Benicia |
CA |
US |
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Prior
Publication: |
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Document Identifier |
Publication Date |
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US 20150306083 A1 |
October 29, 2015 |
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Family ID: |
38521660 |
Appl. No.: |
14/793657 |
Filed: |
July 7, 2015 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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12251310 |
Oct 14, 2008 |
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14793657 |
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PCT/US2007/066544 |
Apr 12, 2007 |
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12251310 |
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60791286 |
Apr 12, 2006 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 36/49 20130101;
A61K 33/24 20130101; A61K 36/54 20130101; A61K 36/22 20130101; A61K
36/38 20130101; A61K 31/4406 20130101; A61K 33/24 20130101; A61K
36/38 20130101; C07C 229/76 20130101; A61K 31/28 20130101; A61K
31/28 20130101; A61K 36/54 20130101; A61K 36/185 20130101; C07C
57/44 20130101; A61K 36/185 20130101; A61K 36/82 20130101; A61K
31/4406 20130101; A61P 7/00 20180101; C07C 65/03 20130101; C07C
59/285 20130101; A61K 36/22 20130101; A61K 36/49 20130101; A61K
36/82 20130101; A61K 2300/00 20130101; A61K 2300/00 20130101; A61K
2300/00 20130101; A61K 2300/00 20130101; A61K 2300/00 20130101;
A61K 2300/00 20130101; A61K 2300/00 20130101; A61K 2300/00
20130101; A61K 2300/00 20130101 |
International
Class: |
A61K 31/4406 20060101
A61K031/4406 |
Claims
1. An isolated and purified complex of trivalent chromium
dinicocysteinate.
2. The isolated and purified complex of claim 1, which is
water-soluble and heat stable.
3. A composition comprising the isolated and purified complex of
claim 1.
4. The composition of claim 3 further comprising a pharmaceutically
acceptable oral, topical, or intravenous carrier or a food or
beverage safe carrier.
5. The composition of claim 4, wherein the composition is in the
form of a pill, a tablet, a capsule, a powder, a lozenge, a gum, a
liquid, a solution, a dietary supplement, a food, a beverage, or a
topical composition.
6. A method for treating or preventing diabetes, the method
comprising administering a biologically effective amount of a
composition comprising an isolated and purified complex of
trivalent chromium dinicocysteinate.
7. The method of claim 6, wherein the biologically effective amount
increases vitamin C levels.
8. The method of claim 6, wherein the biologically effective amount
comprises about 10 .mu.g to about 1000 .mu.g of elemental
chromium.
9. The method of claim 6, wherein the composition is administered
daily.
10. The method of claim 6, wherein the biologically effective
amount comprises about 400 .mu.g of elemental chromium.
11. The method of claim 6, wherein the subject is a mammal.
12. The method of claim 11, wherein the mammal is human.
13. The method of claim 11, wherein the mammal is a swine.
14. The method of claim 6, wherein the subject is avian.
15. A method for treating symptoms associated with diabetes or with
the onset of diabetes, the method comprising administering a
biologically effective amount of a composition comprising an
isolated and purified complex of trivalent chromium
dinicocysteinate to a subject suffering from symptoms associated
with diabetes or with the onset of diabetes.
16. The method of claim 15, wherein symptoms associated with
diabetes or with the onset of diabetes are treated by lowering the
blood levels of one or more of fasting glucose, glycosylated
hemoglobin, monocyte chemotactic protein-1, intracellular adhesion
molecule, and red blood cell lipid peroxidase.
17. The method of claim 15, wherein symptoms associated with
diabetes or with the onset of diabetes are treated by increasing
the blood levels of one or both of vitamin C and adiponectin.
18. A method for treating symptoms associated with inflammation,
the method comprising administering a biologically effective amount
of a composition comprising trivalent chromium dinicocysteinate to
a subject suffering from inflammation.
19. The method of claim 18, wherein symptoms are associated with
atherosclerosis.
20. A method for treating symptoms associated with cardiovascular
disease, the method comprising administering a biologically
effective amount of a composition comprising trivalent chromium
dinicocysteinate to a subject suffering from symptoms associated
with cardiovascular disease.
21. The method of claim 20, wherein symptoms associated with
cardiovascular disease are treated by lowering elevated LDL
cholesterol, VLDL cholesterol, triglycerides, C-reactive protein,
blood pressure and/or vascular inflammation levels, and/or
increasing HDL cholesterol levels.
22. A method for treating symptoms associated with obesity or
overweight individuals, the method comprising administering a
biologically effective amount of a composition comprising trivalent
chromium dinicocysteinate to a subject suffering from symptoms
associated with obesity or overweight individuals.
23. The method of claim 22, wherein symptoms associated with
obesity or overweight individuals are treated by lowering body
weight, body fat and/or body mass index, and/or increasing lean
body mass.
Description
[0001] This application is a divisional which claims benefit to
continuation application Ser. No. 12/251,310, filed Oct. 14, 2008,
which is a continuation-in-part of PCT/US2007/066544, filed Apr.
12, 2007, which claims the benefit of priority from provisional
patent application Ser. No. 60/791,286, filed Apr. 12, 2006, all of
which are incorporated in their entirety by reference herein.
FIELD OF THE INVENTION
[0002] The present invention relates generally to novel trivalent
chromium complexes, which possess the ability to improve insulin
sensitization, glucose tolerance, blood lipid profiles, and lean
body mass through increased chromium bioavailability, efficacy and
safety. More specifically, the present invention relates to
compositions containing trivalent chromium complexes having two
nicotinic acid ligands and a third ligand selected from the group
consisting of alanine, aspartic acid, asparagine, arginine,
cysteine, glutamic acid, glutamine, histidine, isoleucine, lysine,
leucine, methionine, phenylalanine, proline, serine, threonine,
tyrosine, tryptophan, valine, gallic acid, cinnamic acid,
hydroxycitric acid, and 5-hydroxytryptophan with cysteine being
particularly preferred for treatment of various conditions
BACKGROUND OF THE INVENTION
[0003] Although the physiological mechanism is not completely
understood, it has been reported that animals placed on a purified,
chromium-free diet for several weeks displayed greatly impaired
glucose tolerance, i.e. the ability to maintain blood glucose at
normal levels. It was found that a diet containing chromium-rich
Brewer's yeast would eliminate this impairment and that blood
glucose levels would return to normal. The presence of chromium as
an organic salt in foods was also found to increase glucose
oxidation in humans, particularly when extracts of Brewer's yeast
containing chromium where added. In addition, oral administration
of such material to a diabetic individual was found to influence
the pancreas to produce normal amounts of insulin.
[0004] The relationship between chromium content in food and its
effects on glucose oxidation are discussed, for example in Toepfer,
et al., "Chromium Foods in Relation to Biological Activity," J.
Agr. Food. Chem. 21:69 (1973).
[0005] Trivalent chromium (Cr.sup.+3) has long been known to be
essential for proper insulin function and, thus, plays a vital role
in protein, fat and carbohydrate metabolism. In the U.S., studies
show that the diets of nine out of 10 Americans are deficient in
chromium, while chromium levels are known to decline with age.
Stress, exercise and pregnancy are known to increase chromium
losses from the body. Chromium deficiency can lead to symptoms
associated with adult-onset diabetes, obesity and cardiovascular
disease. In studies, supplemental trivalent chromium has improved
blood sugar levels or other symptoms in people with glucose
intolerance, type 1 diabetes, type 2 diabetes, steroid-induced
diabetes and gestational diabetes. Chromium is also known to
increase lean body mass, improve blood lipid profiles and lower
blood pressure levels. However, chromium bioavailability and
biological activity are dependent upon the ligand to which chromium
is bound. (Mertz, W. Chromium Occurrence and Function in Biological
Systems, Physiological Reviews, 49(2): 163-239, 207, 1969 (and see
generally for a discussion of chromium's biological function).
[0006] Inorganic chromium chloride, for example, is poorly absorbed
by the body, typically less than one percent, and has poor
bioavailability. Elucidating the structure, function and mode of
action of the biologically active form of chromium, however, has
proved enigmatic. (Mertz W. Chromium in human nutrition: a review.
J. Nutr. 1993; 123:626-633; Lukaski H. C. Chromium as a supplement.
Annu. Rev. Nutr. 1999; 19:279-301.)
[0007] Various proposals have been developed as to the composition
of a biologically active chromium composition called glucose
tolerance factor (GTF). Walter Mertz has suggested that Brewer's
yeast contained a biologically active form of trivalent chromium
comprised of Cr.sup.3+, glycine, glutamate, cysteine, and nicotinic
acid, which strongly potentiated the action of insulin and possess
an ultraviolet absorbance maximum at about 260 nm. (Schwartz K,
Mertz W: A glucose tolerance factor and its differentiation from
factor 3. Arch Biochem Biophys 72: 515-518, 1957; Toepfer E W,
Mertz W, Polansky M M, Roginski E E, Wolfe W R: Preparation of
chromium containing material of glucose tolerance factor activity
from Brewer's yeast extracts and by synthesis. J. Agricul Food Chem
25: 162-166, 1977.) However, extracting the biologically active
fractions of chromium in Brewer's yeast involves complex processes,
which are expensive on a commercial scale for use in chromium
supplementation of chromium deficient diets or for individuals
otherwise requiring chromium supplementation. Therefore,
researchers have tried to overcome this problem by synthetically
preparing biologically active chromium compounds. In this regard,
see Cooper et al., Inorganica Chimica Acta 92:23-31 (1984), which
is directed to the synthesis and analysis of possible components of
GTF. In Cooper, various complexes of glycine, glutamic acid,
glutamine, cysteine and nicotinic acid were formed and analyzed for
glucose tolerance factor activity using a yeast assay. A number of
complexes including Cr-(cysteine).sub.2.sup.-, Cr-nicotinic
acid-cysteine complexes of undetermined structure, and Cr-nicotinic
acid glycine were tested but only
Cr(glutamine).sub.2(H.sub.2O).sub.2.sup.+, Cr-nicotinic
acid-glycine and the mixture of complexes of
Cr(glycine).sub.n(H.sub.2O).sub.6-n.sup.+3 showed significant
activity. In contrast, Cr(cysteine).sub.2.sup.- and
Cr-nicotinic-cysteine complexes were not active in the yeast
bioassay.
[0008] Other chromium complexes have been proposed as well.
Maciejewska et al., Transition Metal Chem. 27:473-480 (2002) tested
the toxicity and determined structure data of seven mono- and
poly-nuclear Cr.sup.III complexes with natural ligands: glycine,
glutaminic, nicotinic and asparginic acid, cysteine, and
glutathione. Gonzales-Vergara et al., Israel J. Chem. 21:18-22
(1981) synthesized CrCl.sub.3, CrEDTA, Cr(glycine).sub.2(nicotinic
acid).sub.2, Cr(III)-pyridoxylideneglycylglycine-diaquo
(Cr(III)-PGG-diaquo), and Cr(III)-PGG-(nicotinic acid).sub.2, which
were tested for retention in mice.
[0009] The importance of the B vitamin, niacin, to chromium's
biological activity was elucidated by Walter Mertz, when it was
discovered that chromium bound to niacin strongly potentiated the
action of insulin in vitro, while chromium alone, niacin alone or
chromium bound to isomers of niacin, including picolinic acid, had
virtually no effect on insulin in vitro. (Mertz W, Effects and
Metabolism of Glucose Tolerance Factor, Present Knowledge in
Nutrition, 4.sup.th Edition, The Nutrition Foundation, Washington,
D.C., Chapter 36, pp 365-372, 1976.)
[0010] Niacin can be bound to chromium in various configurations. A
particular oxygen-coordinated niacin-bound chromium complex was
developed and introduced commercially as ChromeMate.RTM.
(Interhealth Nutraceuticals, Inc., Benecia, Calif.). (Jensen, U.S.
Pat. No. 5,194,615). Subsequently, Cooper et al. determined that
oxygen-coordinated niacin-bound chromium was up to 18-times more
potent than other forms of niacin-bound chromium tested in vitro.
(Cooper J, et al, Structure and Biological Activity of Nitrogen and
Oxygen Coordinated Nicotinic Acid Complexes of Chromium, Inorganica
Chimica Acta, 91:1-9, 1984.)
[0011] Further studies established the superior safety and efficacy
of ChromeMate.RTM. over inorganic chromium chloride and chromium
picolinate, two commercially available forms of trivalent chromium
for dietary supplementation. (Jain S K, Rains J and Rogier K,
Effect of Niacin-Bound Chromium Complex (NBC) on IL-6 Secretion and
Oxidative Stress Caused by High Glucose (HC) in Cultured U397
Monocytes, FASEB, 20(4):132, Abs. 376.4, April 2006; Grant K E,
Chandler R M, Castle A L, Ivy J L, Chromium and Exercise Training:
Effect on Obese Women, Medicine & Science in Sports &
Exercise, 29:992-998, 1997; Preuss H G, Gropec P L, Lieberman S and
Anderson R A, Effects of Different Chromium Compounds on Blood
Pressure and Lipid Peroxidation in Spontaneously Hypertensive Rats,
Clinical Nephrology, 47:325-330, 1997; Stearns D M, Wise, Sr., J P,
Patierno S R and Wetterhahn K E, Chromium (III) Picolinate Produces
Chromosome Damage in Chinese Hamster Ovary Cells. The FASEB
Journal, 9: 643-1648, 1995.) In human bioavailability equivalency
studies, researchers at U. C. Davis demonstrated that
ChromeMate.RTM. was 311% more bioavailable than chromium picolinate
and 672% more than chromium chloride in animal models. (Olin et
al., Comparative retention/absorption of .sup.51chromium
(.sup.51Cr) from .sup.51Cr chloride, .sup.51Cr nicotinate and
.sup.51Cr picolinate in a rat model, Trace Elements and
Electrolytes, 11(4):182-186, 1994.)
[0012] John Vincent has proposed a naturally occurring
oligopeptide, low-molecular-weight chromium-binding substance
(LMWCr) or chromodulin. Chromodulin has been proposed to activate
insulin receptor kinase activity. The oligopeptide possesses a
molecular weight of 1500 Da and is comprised of four types of amino
acid residues: glycine, cysteine, glutamate and aspartate. (Vincent
J. B: The quest for the molecular mechanism of chromium action and
its relationship to diabetes. Nutr. Rev. 58, 2000.)
[0013] Taken together, the ligands of four amino acids, glycine,
cysteine, glutamate and aspartate, and niacin are important for
bioactive chromium complexes. (Yamamoto A., Wada O, Ono T:
Isolation of a biologically active low-molecular-mass chromium
compound from rabbit liver. Eur. J. Biochem. 165: 627-631, 1987;
Davis C M, Vincent J B, Chromium oligopeptide activates insulin
receptor tyrosine kinase activity. Biochemistry 36: 4382-4385,
1997.)
[0014] Another proposal supports the use of the amino acid
histidine for GTF activity, U.S. Pat. No. 6,689,383, which is
incorporated by reference in its entirety. Chromium histidine is
said to be absorbed at least 50 percent better than chromium
picolinate. In tests, men and women absorbed an average 3.1 .mu.g
of chromium from the chromium-histidine complex, compared with 1.8
.mu.g from chromium picolinate, 0.4 .mu.g from chromium chloride
and 0.2 .mu.g from chromium polynicotinate.
[0015] Alternatively, Yang et al. have shown the use of
triphenylalanine as a ligand for a bioactive form of chromium.
(Yang X, Palanichamy K, Ontko A C, Rao M N A, Fang C, Ren J,
Sreejayan N: A newly synthetic chromium complex--chromium
triphenylalanine improves insulin responsiveness and reduces whole
body glucose tolerance, FEBS Letters 579 1458-1464, 2005.)
[0016] Synthesis of chromium amino acid nicotinate complex with
mixture of glycine, glutamic acid and cysteine has been disclosed
in U.S. Pat. No. 5,536,838, which is incorporated by reference in
its entirety.
[0017] Chromium (III) 1:3 complexes of alpha amino acids are
disclosed as animal nutritional supplements in Abdel-Monem et al.,
U.S. Pat. No. 7,247,328. Exemplified amino acids are methionine and
leucine.
[0018] Gillota, U.S. Patent Publication No. 2003/0143311, discloses
a recovery drink formula that includes chromium in the form of
chromium dinicotinate glycinate.
[0019] In addition to a desire for safety, efficacy and
bioavailability, there remains a desire for stability, solubility
and the effect on taste and odor for use in food and beverage
applications. Thus, while there have been many proposed chromium
complexes for use in dietary supplements and food and beverage
applications, most are inadequate or poorly characterized, and
there still remains a need in the art for an improved synthetic
chromium compound, which demonstrates improved biological activity,
bioavailability, stability, solubility and/or sensory
characteristics.
SUMMARY OF THE INVENTION
[0020] The present invention relates to the discovery that chromium
dinicocysteinate is characterized by improved physical properties
and by increased chromium bioavailability and efficacy once
administered to a subject. Chromium dinicocysteinate is a trivalent
chromium complex with two nicotinic acid ligands and one cysteine
ligand, also known as chromium dinicotinocysteinate, dinicotino
cysteinato chromium complex, or chromium dinicotinate cysteinate.
In addition to chromium dinicocysteinate, disclosed herein are
various other chromium complexes, compositions comprising chromium
complexes, and methods of using these complexes to promote or
improve various health conditions and functions of the body.
[0021] Methods of administering these complexes to treat or prevent
conditions including but not limited to diabetes, symptoms
associated with diabetes, and/or the onset of diabetes include
administering a biologically effective amount of chromium
dinicocysteinate. Chromium dinicocysteinate demonstrates reductions
in fasting glucose levels, glycated hemoglobin, C-reactive protein,
monocyte chemotactic protein-1, intracellular adhesion molecule,
and red blood cell lipid peroxidation. In addition, it demonstrated
elevation of adiponectin and vitamin C levels, which reduces the
risk of developing diabetes. Preferably, the biologically effective
amount is about 10 .mu.g to about 1000 .mu.g of elemental chromium,
and more preferably, 400 .mu.g of elemental chromium. The methods
can further comprise treating the symptoms by lowering the blood
levels of one or more of fasting glucose, glycosylated hemoglobin,
C-reactive protein, monocyte chemotactic protein-1, intracellular
adhesion molecule, and red blood cell lipid peroxidase, or by
increasing the blood levels of one or both of vitamin C and
adiponectin.
[0022] Disclosed herein are methods directed to treating the
symptoms associated with cardiovascular disease including
administering a biologically effective amount of chromium
dinicocysteinate. Cardiovascular disease can be treated by lowering
elevated LDL cholesterol, VLDL cholesterol, triglycerides,
C-reactive protein, blood pressure, and/or vascular inflammation
levels, or by increasing HDL cholesterol levels. For example, the
disclosed complexes, particularly chromium dinicocysteinate,
demonstrate reduction in C-reactive protein, which correlates to
cardiovascular disease.
[0023] Disclosed herein are methods directed to treating symptoms
associated with obesity and overweight individuals including
administering a biologically effective amount of chromium
dinicocysteinate. Symptoms associated with obesity and overweight
individuals can be treated by lowering body weight, body fat,
and/or body mass index, and/or increasing lean body mass. For
example, the disclosed complexes, particularly chromium
dinicocysteinate, demonstrate a reduction in food intake.
[0024] In various embodiments, the subject is a mammal. In specific
embodiments, the subject is human or swine. In alternative
embodiments, the subject is avian.
[0025] In still another aspect, disclosed herein are compositions
including the disclosed chromium complexes. The compositions can
include a pharmaceutically acceptable carrier or a food safe
carrier. In some embodiments, the amount of chromium complex in the
composition is about 0.000001% to about 1% by weight of the total
composition. In various embodiments, the compositions disclosed
herein are formulated as a pill, tablet, capsule, powder, lozenge,
gum, liquid, solution, dietary supplement, food, beverage, or
topical composition.
[0026] Novel chromium compounds, which improve insulin
sensitization, glucose metabolism or tolerance, blood lipid
metabolism, bioavailability and/or safety as compared to chromium
nicotinate, chromium picolinate and chromium histidine are
disclosed. These compounds were evaluated in terms of solubility,
in vitro insulin sensitization, stability, and absorption as
compared to chromium nicotinate, chromium picolinate and chromium
histidine. The most promising chromium compounds have been tested
for their animal in vivo bioavailability, glucose metabolism, lipid
metabolism and acute toxicity.
[0027] In another embodiment, disclosed herein are chromium
compounds (alternatively called chromium complexes) include
dinicotinate chromium complexes having a carboxylate ligand as a
third ligand. The third ligand can be selected from the group
consisting of alanine, aspartic acid, asparagine, arginine,
cysteine, glutamic acid, glutamine, histidine, isoleucine, lysine,
leucine, methionine, phenylalanine, proline, serine, threonine,
tyrosine, tryptophan, valine, gallic acid, cinnamic acid,
hydroxycitric acid, and 5-hydroxytryptophan. These complexes can be
administered to treat or prevent diabetes, the symptoms associated
with diabetes, and/or the onset of diabetes. These complexes can be
administered to treat the symptoms associated with cardiovascular
disease. In addition, these complexes can be administered to treat
associated with obesity and overweight individuals. The third
ligand can be a simple carboxylate (e.g., propionate), a
carboxylate having other coordinating functionality (e.g., amino
acids or hydroxy-acids), and/or a carboxylate having pi electrons
available for potential secondary bonding to chromium through
chromium's d-orbitals (e.g., tyrosine or tryptophan).
[0028] Additional features of the invention may become apparent to
those skilled in the art from a review of the following detailed
description, taken in conjunction with the drawings, the examples,
and the appended claims.
DETAILED DESCRIPTION OF THE INVENTION
[0029] Chromium complexes disclosed herein are complexes of
chromium having two nicotinic acid ligands and a third ligand
having a carboxylate functional group, such as glutamate, cysteine,
aspartate, tryptophan, cinnamate, and the like. Preferred
compositions include an amino acid as the third ligand, preferably
cysteine. These complexes have been compared in various
biologically assays for their ability to increase and/or influence
the bioavailability and/or biological activity of chromium.
[0030] In an embodiment, disclosed herein are complexes of chromium
and one, two, or three ligands selected from the group consisting
of glycine, alanine, aspartic acid, asparagines, arginine,
cysteine, glutamic acid, glutamine, histidine, isoleucine, lysine,
leucine, methionine, phenylalanine, praline, serine, threonine,
tyrosine, tryptophan, valine, gallic acid, cinnamic acid,
hydroxycitric acid, 5-hydroxytrytophan, and nicotinic acid.
[0031] In various cases, the ligand(s) has/have the ability to bond
to chromium via its carboxylate functional group as well as through
pi electron-d orbital interaction. This secondary interaction
between the ligand and chromium can increase the bioavailability
and absorption of chromium.
[0032] In still another aspect, the chromium complexes are
complexes of trivalent chromium and at least one and no more than
three tyrosine or tryptophan ligands. In specific embodiments, the
present invention provides chromium complexes such as chromium
(III) tris(tryptophan) and chromium (III) tris(tyrosine).
[0033] In another aspect, the chromium complexes are complexes of
trivalent chromium and one or more compounds extracted from plants.
The plants from which these compounds are extracted are typically
plants shown to have beneficial health benefits, such as genus
Garcinia, Groffonia simplicifolia, cinnamon bark, gallnuts, sumac,
witch hazel, tea leaves, and oak bark. In specific embodiments, the
present invention provides chromium complexes such as chromium
hydroxycitrate, chromium hydroxytryptophan, chromium cinnamate, and
chromium gallate.
[0034] In some cases, the ligand has known biological function
alone. Examples include, but are not limited to, hydroxycitric
acid, 5-hydroxytryptophan, gallate, and cinnamate.
[0035] In yet another aspect, methods are disclosed for improving
or promoting healthy insulin function, glucose tolerance, blood
sugar levels, blood lipid levels, blood pressure levels,
inflammation, and/or lean body mass comprising administering a
biologically effective amount of a composition as disclosed herein
to a subject in need thereof, wherein the composition provides
sufficient chromium to the subject and improves or promotes one or
more of the identified health conditions, which are now known or
are discovered to be responsive to chromium levels in the
subject.
Inflammation
[0036] It is contemplated that the disclosed compounds are useful
for treating symptoms associated with inflammation. Inflammation is
the complex biological response of vascular tissues to harmful
stimuli, such as pathogens, damaged cells, or irritants. It is a
protective attempt by the organism to remove the injurious stimuli
as well as initiate the healing process for the tissue.
[0037] Chronically inflamed tissue is characterized by the
infiltration of mononuclear immune cells, including monocytes,
macrophages, lymphocytes, and plasma cells. Nuclear factor kappa B
(NF-.kappa.B) also plays an important role in inflammation through
modulation of proinflammatory cytokines including, for example,
interleukin 6 (IL-6), interleukin 17A (IL-17A), and tumor necrosis
factor-alpha (TNF.alpha.). Increased levels of monocyte chemotactic
protein-1 (MCP-1), C-reactive protein (CRP), and oxidative stress
and decreased levels of adiponectin are known to increase vascular
inflammation. The compounds disclosed herein, specifically chromium
dinicocysteinate for example, demonstrate reductions in CRP and
MCP-1, and increases in adiponectin, and are contemplated to be
useful in the treatment of inflammatory conditions, such as, hay
fever, rheumatoid arthritis, and atherosclerosis, which is a
chronic inflammatory response in the wall of arteries.
Reduced Oxidative Stress
[0038] According to another embodiment, it is contemplated that the
disclosed compounds are useful for reducing oxidative stress.
Inflammation and oxidative stress are closely linked. Inflammation
and oxidative stress are usually observed in degenerative or
chronic diseases. The ZDF rat is a model of type 2 diabetes and is
associated with elevated levels of both hyperglycemia and
pro-inflammatory cytokines. Hyperglycemia causes excessive oxygen
radical production leading to increased oxidative stress in
diabetes (Evans et al.: The molecular basis for oxidative
stress-induced insulin resistance. Antioxidant Redox Signaling.
7:1040-1052, 2005). Oxidative stress also influences the expression
of multiple genes in vascular cells, including signaling molecules
such as protein kinases (PK) and nuclear factor kappa B
(NF-.kappa.B). Over-expression of these genes may induce the
secretion of pro-inflammatory cytokines. Oxidative stress plays a
key role in a cascade of events which results in elevated glucose
leading to monocyte and endothelial cell activation and finally the
enhanced vascular inflammation observed in diabetes (Guha et al.:
Molecular mechanisms of tumor necrosis factor a gene expression in
monocytic cells via hyperglycemia-induced oxidant stress-dependent
and -independent pathways. Journal of Biological Chemistry.
275:17728-17739, 2000). For example, chromium dinicocysteinate
reduced elevated blood lipid peroxidation levels in Zucker diabetic
fatty (ZDF) rats. Without intending to be bound by theory, it is
believed that the underlying molecular mechanism of the observed
beneficial effects of chromium dinicocysteinate are likely to be
due to the inhibition of oxidative stress-signaling molecules such
PKB and NF-.kappa.B.
Improved Liver and Kidney Function
[0039] In one embodiment, it is contemplated that the disclosed
compounds improve liver and/or kidney function. A recent study
showed an inverse correlation of insulin sensitivity to elevated
alkaline phosphatase activities in the blood of 472 apparently
healthy men (Godsland and Johnston: Co-associations between insulin
sensitivity and measures of liver function, subclinical
inflammation, and hematology. Metabolism Clinical and Experimental.
57:1190-7, 2008). The liver plays an important role in maintaining
normal glucose concentrations during fasting as well as
postprandial. Insulin dysfunction leads to ineffective suppression
of hepatic glucose production and glycogenolysis in the liver
causing an increase in hepatic glucose production (Michael and
Kulkarni: Loss of Insulin signaling in hepatocytes leads to severe
insulin resistance and progressive hepatic dysfunction. Molecular
Cell. 6:87-97, 2000). Evidence also indicated that liver function
and subclinical inflammation are related to insulin sensitivity
which predict new-onset of type 2 diabetes independent of classical
risk factors (Hsiao et al.: Decreased Insulin Secretion and Insulin
Sensitivity Are Associated With Liver Function in Subjects With
Fasting Glucose Between 100 and 109 mg/dL in Taiwanese Population.
Pancreas 35:343-7, 2008). Without intending to be bound by theory,
it is believed that a decrease in the alkaline phosphatase
activities in the chromium dinicocysteinate-supplemented group
suggests an improvement in liver function which may play a role in
the improved glycemia-related parameters as observed in the
chromium dinicocysteinate-supplemented Zucker diabetic fatty rats.
For example, chromium dinicocysteinate reduces alkaline phosphatase
activities. In addition, chromium dinicocysteinate can improve
kidney and/or endothelial cellular functions by reducing blood
creatinine.
Compositions of Chromium Complexes
[0040] The chromium complexes disclosed herein can be incorporated
into a composition. These compositions can further comprise a
pharmaceutically acceptable carrier or excipient or a food safe
carrier or excipient. The amount of the chromium complex is
typically about 0.000001% to about 1% by weight of the total
composition. The chromium complex can be about 0.00001% to about
0.5%; about 0.00001% to about 0.1%; about 0.001% to about 0.5%, or
about 0.001% to about 0.01% by weight.
[0041] The compositions disclosed herein typically provide a total
amount of chromium upon administration to a subject in need of
about 10 .mu.g to about 1000 .mu.g. The amount of chromium provided
can be about 20 .mu.g to about 500 .mu.g; about 20 .mu.g to about
250 .mu.g; about 100 .mu.g to about 750 .mu.g; about 250 .mu.g to
about 750 .mu.g; or about 500 .mu.g to about 1000 .mu.g.
[0042] An "effective amount" of the disclosed complexes refers to
the amount or quantity of the complex, which is sufficient to
elicit the required or desired prophylactic or therapeutic
response, or in other words, the amount which is sufficient to
elicit an appreciable biological response when administered to a
subject. For example, an effective amount of a chromium complex as
disclosed herein may reduce blood sugar levels in a subject. The
actual effective amount for a particular subject can be readily
determined by a person of ordinary skill in the art by various
dosing tests and evaluations of chromium complexes for their effect
on, for instance, fasting glucose levels, blood vitamin C levels,
and the like. For example, in the study disclosed in the Example
section below, a biologically effective amount for obese Zucker
rats was 400 .mu.g per kilogram body weight.
[0043] The compositions disclosed herein comprise a
pharmaceutically acceptable carrier or a food safe carrier. Such
carriers can be those that are approved by regulatory agencies,
such as the U.S. Food and Drug Administration and its national
counterparts or those carriers generally regarded as safe (GRAS) in
the relevant industry. Exemplary carriers include phosphate
buffered saline solution, and 5% aqueous solution of dextrose. The
present compositions can be in the form of an emulsion, such as an
oil/water or water/oil emulsion, and various types of wetting
agents and/or adjuvants, that are approvable by a competent
regulatory authority as suitable for administration. Suitable
pharmaceutical carriers and formulations are described in
Remington's Pharmaceutical Sciences, 19th Ed. (Mack Publishing Co.,
Easton, 1995).
[0044] Preferred pharmaceutical carriers depend upon the intended
mode of administration of the active agent. Typical modes of
administration include enteral (e.g., oral), topical, or parenteral
(e.g., subcutaneous, intramuscular, intravenous, intraperitoneal or
intrathecal injection; transdermal, or transmucosal, including
intrapulmonary administration). Pharmaceutically acceptable
ingredients are well known for the various types of compositions
and may be, for example, binders such as natural or synthetic
polymers, excipients, lubricants, surfactants, sweetening and
flavoring agents, coating materials, preservatives, dyes,
thickeners, adjuvants, antimicrobial agents, antioxidants and
carriers for the various formulation types. Nonlimiting examples of
binders useful in a composition described herein include gum
tragacanth, acacia, starch, gelatin, and biological degradable
polymers such as homo- or co-polyesters of dicarboxylic acids,
alkylene glycols, polyalkylene glycols and/or aliphatic hydroxyl
carboxylic acids; homo- or co-polyamides of dicarboxylic acids,
alkylene diamines, and/or aliphatic amino carboxylic acids;
corresponding polyester-polyamide-co-polymers, polyanhydrides,
polyorthoesters, polyphosphazene and polycarbonates. The biological
degradable polymers may be linear, branched or crosslinked.
Specific examples are poly-glycolic acid, poly-lactic acid, and
poly-d,l-lactide/glycolide. Other examples for polymers are
water-soluble polymers such as polyoxaalkylenes (polyoxaethylene,
polyoxapropylene and mixed polymers thereof, poly-acrylamides and
hydroxylalkylated polyacrylamides, poly-maleic acid and esters or
-amides thereof, poly-acrylic acid and esters or -amides thereof,
poly-vinylalcohol and esters or -ethers thereof,
poly-vinylimidazole, poly-vinylpyrrolidon, and natural polymers
like chitosan.
[0045] Nonlimiting examples of excipients useful in a composition
described herein include phosphates such as dicalcium phosphate.
Nonlimiting examples of lubricants use in a composition described
herein include natural or synthetic oils, fats, waxes, or fatty
acid salts such as magnesium stearate.
[0046] Surfactants for use in a composition described herein can be
anionic, cationic, amphoteric or neutral. Nonlimiting examples of
surfactants useful in a composition described herein include
lecithin, phospholipids, octyl sulfate, decyl sulfate, dodecyl
sulfate, tetradecyl sulfate, hexadecyl sulfate and octadecyl
sulfate, Na oleate or Na caprate, 1-acylaminoethane-2-sulfonic
acids, such as 1-octanoylaminoethane-2-sulfonic acid,
1-decanoylaminoethane-2-sulfonic acid,
1-dodecanoylaminoethane-2-sulfonic acid,
1-tetradecanoylaminoethane-2-sulfonic acid,
1-hexadecanoylaminoethane-2-sulfonic acid, and
1-octadecanoylaminoethane-2-sulfonic acid, and taurocholic acid and
taurodeoxycholic acid, bile acids and their salts, such as cholic
acid, deoxycholic acid and sodium glycocholates, sodium caprate or
sodium laurate, sodium oleate, sodium lauryl sulphate, sodium cetyl
sulphate, sulfated castor oil and sodium dioctylsulfosuccinate,
cocamidopropylbetaine and laurylbetaine, fatty alcohols,
cholesterols, glycerol mono- or -distearate, glycerol mono- or
-dioleate and glycerol mono- or -dipalmitate, and polyoxyethylene
stearate.
[0047] Nonlimiting examples of sweetening agents useful in a
composition described herein include sucrose, fructose, lactose,
aspartame, saccharine, acesulfame potassium, or sucralose.
Nonlimiting examples of flavoring agents for use in a composition
described herein include peppermint, oil of wintergreen or fruit
flavors such as cherry or orange flavor. Nonlimiting examples of
coating materials for use in a composition described herein include
gelatin, wax, shellac, sugar or other biological degradable
polymers. Nonlimiting examples of preservatives for use in a
composition described herein include methyl or propylparabens,
sorbic acid, chlorobutanol, phenol and thimerosal.
[0048] Additionally or alternatively, the present chromium
compositions can be incorporated into food or beverages. Juices,
energy drinks, diet drinks, protein bars, and the like can be used
as a vehicle for oral delivery of the present chromium
compositions.
Screening of Novel Chromium Complexes
[0049] Three phases of screening of chromium compounds were
conducted. Phase I included synthesis and chemical/physical
characterization of the novel chromium complexes in comparison to
know (reference or control) compounds. Phased II included in vitro
insulin sensitization, absorption and stability tests on all of the
synthesized chromium complexes as compared to reference or control
compounds, such as chromium nicotinate, chromium picolinate,
chromium histidine and chromium triphenylalanine. Phase III
included in vivo bioavailability, glucose metabolism, lipid
metabolism and acute toxicity.
[0050] Still another set of chromium complexes disclosed herein are
chromium having three different carboxylate ligands. By varying
ligands from nicotinic acid, glutamate, cysteinate, aspartate,
argininate, tyrosine and tryptophan, at least 30 possible chromium
complexes are produced. Each of these compounds can be assessed for
their ability to provide chromium in a bioavailable form using the
assays disclosed herein.
[0051] Some chromium compounds contemplated include, but are not
limited to, the following: Chromium dinicocysteinate; Chromium
dinicotinate tryptophan; Chromium dinicotinate tyrosine; Chromium
dinicotinate hydroxycitrate; Chromium dinicotinate cinnamate;
Chromium dinicotinate gallate; Chromium dinicotinate
5-hydroxytryptophan; Chromium dinicotinate aspartate; Chromium
dinicotinate glutamate; Chromium dinicotinate arginate; Chromium
tris(tryptophan); Chromium tris(tyrosine); Chromium
tris(hydroxycitrate); Chromium tris(5-hydroxytryptophan); Chromium
tris(cinnamate); and Chromium tris(gallate).
Chromium Hydroxycitrate
[0052] Hydroxycitric acid (HCA) is a naturally occurring acid found
in the rinds of the fruit of Garcinia cambogia, Garcinia indica,
Garcinia mangostana, and Garcinia atrovirides. The dried fruit rind
of G. cambogia, also known as Malabar tamarind, is commonly used in
Southeast Asia (particularly southern India) as a food
preservative, flavoring agent and carminative. The primary
mechanism of action of (-)-HCA appears to be related to act as a
competitive inhibitor of the enzyme ATP-citrate lyase, which
catalyzes the conversion of citrate and coenzyme A to oxaloacetate
and acetyl coenzyme A (acetyl-CoA), building blocks of fatty acid
synthesis. Extensive experimental studies suggest that (-)-HCA
suppresses fatty acid synthesis, lipogenesis and food intake, thus
leading to weight reduction. In addition to suppression of fatty
acid and fat synthesis, (-)-HCA is thought to suppress food intake
via loss of appetite by stimulation of liver gluconeogenesis.
Various researchers have evaluated HCA for its weight control
properties, fat burning properties, lipid level lowering effect,
appetite regulation, metabolic rate increase and other effects. A
number of patents have been granted based on the above studies and
various methods of extraction of HCA from the fruit. The isolation
and chemical nature of (-)-HCA from Garcinia rind are well studied.
(See, e.g., Lewis, Y. S. et al, Phytochemistry, 4, 619-625, 1965;
U.S. Pat. No. 5,656,314, which is incorporated by reference in its
entirety.) Recent literature reveals that (-)-HCA acts as a glucose
absorption regulator. (See, e.g., Wielinga, Y. P. et al.
Hydroxycitric acid delays glucose absorption in rats, Am. J.
Physiol. Gastrointest. Liver Physiol., 288:1144-1149, 2005). Since
HCA alone has been shown to have beneficial effects in regulating
glucose absorption and in weight control, HCA complexed to chromium
provides a doubly beneficial effect--through the effect of HCA and
through the effect of bioavailable chromium.
Chromium Cinnamate or Chromium Gallate
[0053] Cinnamic acids are widely distributed in plant kingdom and
are reported as antioxidants. These compounds impart nutraceutical
traits to foods by way of their abilities to serve as cellular
antioxidants, anti-inflammatory agents or inhibitors of enzymes
involved in cell proliferation. These activities are important in
ameliorating chronic diseases such as cancer, arthritis and
cardiovascular disease, which in some cases may be caused by free
radicals. Because of proven safety of cinnamic acids like caffeic
acid and phenolic benzoic acids like gallic acid, chromium
complexes of cinnamic acid and gallic acid may have insulin
sensitizing capacity.
Chromium 5-Hydroxytryptophan Complex
[0054] 5-Hydroxytryptophan (5-HTP) is a natural compound isolated
from the seeds of an African plant called Griffonia simplicifolia.
Substances which increase brain serotonin (5-hydroxytryptamine,
5-HT) are effective anorectic agents to help obese patients lose
weight and to decrease cravings for sweets and carbohydrates.
Experimental studies have proven that 5-HTP increases brain
serotonin (5-HT), a neurotransmitter involved in appetite control,
sleep and mood. Chromium complexes of 5-hydroxytryptophan may
exhibit these same beneficial effects in addition to the beneficial
effects due to bioavailable chromium.
Synthesis and Characterization of Chromium Complexes and Reference
Compounds
[0055] Synthesis and characterization of chromium complexes
disclosed herein is performed according to the methods disclosed in
the examples below.
[0056] Chromium compounds were screened employing the following in
vitro assays as compared to known chromium complexes: chromium
nicotinate, chromium picolinate, chromium histidine, chromium
triphenylalaine and chromium nicotinoglycinate: 1. Solubility
analysis; 2. Insulin sensitivity and glucose metabolism/utilization
assay; 3. Stability analysis (time, temperature and pH); 4.
Cytotoxicity assay (lactate dehydrogenese leakage assessment); 5.
Absorption study using the intestinal reperfusion model.
[0057] The most promising compounds were tested employing the
following assays as compared to other chromium complexes: chromium
nicotinate, chromium picolinate, chromium histidine, chromium
triphenylalaine and chromium nicotinoglycinate: 1. Bioavailability;
2. Glucose metabolism/uptake/utilization; 3. Insulin sensitivity;
4. Lipid metabolism (animal in vivo); 5. Acute toxicity
(LD.sub.50).
EXAMPLES
Tryptophan-Chromium Complex
Method I
[0058] The synthesis was made effective through an intermediate of
tris-(ethylenediamine) chromium.
Synthesis of tris(ethylenediamine)chromium.
[0059] Zinc metal (1 g) was added to a solution of chromium(III)
chloride (CrCl.sub.3.6H.sub.2O, 26.6 g, 0.11 mole) in methanol (50
mL). The mixture was refluxed at 70-80.degree. C. Anhydrous
ethylenediamine (36 g, 0.6 mole) was added drop-wise to the
refluxing chromium salt solution and the refluxing was continued
for one hour. The solution was cooled, filtered and the yellow
product, washed first with 10% ethylenediamine solution in methanol
(75 mL) and then with ether. Finally the product was dried in air
(yield 19 g).
Synthesis of Tris(Tryptohanato)Chromium
[0060] Tris(ethylenediamine)chromium (0.6 g, 1.9 mmol) was added to
a stirred suspension of hot L-tryptophan (2.0 g, 9.6 mmol) in water
(30 mL). The mixture was heated at 80-90.degree. C. refluxed
further for 5 hour and the solid product obtained was filtered
while hot. The solid was washed with hot water (200 mL) to remove
the unreacted starting materials. The product was vacuum dried (0.8
g, 64%). The Electrospray Ionization Mass Spectrum of the compound
indicated the peak corresponding to the product.
Method II
[0061] A suspension of tryptophan (4.59 g, 22.5 mmol) in water (120
mL) was heated to 70-80.degree. C. A solution of chromium chloride
(CrCl.sub.3.6H.sub.2O, 1.995 g, 7.5 mmol) in water (15 mL) was
added to it and was maintained at this temperature under stirring
for 7 h. The resulting mixture was cooled to room temperature,
frozen at -80.degree. C. and lyophilized. After lyophilizing for 48
h, the dark purple solid was washed with acetone and air dried to
obtain the pure product. The product was subjected to elemental
analysis: Calculated for Cr(TRP).sub.3.3HCl.2H.sub.2O: C, 49.11; H,
5.00; N, 10.41; Observed: C, 49.84; H, 5.18; N, 10.41. Electro
Spray Ionization Mass Spectrometry: Calculated for Cr(TRP).sub.3:
661; Observed a peak at 662.3 (M+1)
Tyrosine-Chromium Complex
Method I
[0062] Due to poor solubility of Tyrosine, even in DMSO, the method
I using Cr(en)3 complex did not work well, therefore the second
method was adopted for the synthesis.
Method II
[0063] A suspension of tyrosine (8.145 g, 44.95 mmol) in water (350
mL) was heated to reflux. A solution of Chromium chloride
(CrCl.sub.3.6H.sub.2O, 2.66 g, 10 mmol) in DI water (35 mL) was
added to it and was maintained refluxing overnight, under stirring.
Cooled to room temperature, and filtered the purplish solution
(containing lot of un-dissolved white material). The solution was
frozen at -80.degree. C. and lyophilized. After lyophilizing for 48
h, the dark purple solid was washed with acetone and finally dried
in air to get the pure product. Elemental analysis: Calculated for
Cr(TYR).sub.3.3HCl.7H.sub.2O: C, 39.16; H, 5.72; N, 5.07; Observed:
C, 39.59; H, 5.12; N, 4.33. Electro Spray Ionization Mass:
Calculated for Cr(TYR).sub.3 592; Observed a peak at 593 (M+1).
5-Hydroxytryptophan Chromium Complex
[0064] A suspension of 5-Hydroxytryptophan (2.20 g, 10 mmol) in DI
water (100 mL) was heated to 70-80.degree. C. A solution of
Chromium chloride (CrCl.sub.3.6H.sub.2O, 0.88 g, 3.31 mmol) in DI
water (20 mL) was added to it and was maintained at this
temperature under stirring for 9 h. Cooled to room temperature,
frozen at -80.degree. C. and lyophilized. After lyophilizing for 48
h, the dark purple solid was washed with acetone and finally dried
in air to get the pure product. Elemental analysis: Calculated for
Cr(HTRP).sub.3.3HCl.2H.sub.2O: C, 46.35; H, 4.72; N, 9.83;
Observed: C, 46.17; H, 4.84; N, 9.53. Electro Spray Ionization
Mass: Calculated for Cr(TYR).sub.3 709; Observed a peak at 710
(M+1).
Dinicotino Glutamino Chromium Complex
[0065] A suspension of nicotinic acid (3.60 g, 29.24 mmol) in DI
water (120 mL) was heated to 70-80.degree. C. Glutamic acid (2.22
g, 15.1 mmol) was added to it and continued heating. Once the whole
material was dissolved a solution of Chromium chloride
(CrCl.sub.3.6H.sub.2O, 4.05 g, 15.2 mmol) in DI water (20 mL) was
added to it. The mixture was stirred at 70-80.degree. C. for 9 h
and cooled to room temperature, frozen at -80.degree. C. and
lyophilized. The dark solid obtained was collected and washed with
acetone and dried to get the product. Elemental analysis:
Calculated for Cr(NIC).sub.2(GLU).3HCl.3H.sub.2O: C, 33.73; H,
4.16; N, 6.94; Observed: C, 33.40; H, 3.70; N, 6.91.
Dinicotino Aspartato Chromium Complex
[0066] A suspension of nicotinic acid (3.60 g, 29.24 mmol) in DI
water (120 mL) was heated to 70-80.degree. C. Aspartic acid (1.99
g, 15.1 mmol) was added to it and continued heating. Once the whole
material was dissolved a solution of Chromium chloride
(CrCl.sub.3.6H.sub.2O, 4.05 g, 15.2 mmol) in DI water (20 mL) was
added to it. The mixture was stirred at 70-80.degree. C. for 9 h
and cooled to room temperature, frozen at -80.degree. C. and
lyophilized. The dark solid obtained was collected and washed with
acetone and dried to get the product.
Chromium Dinicocysteinate Complex
[0067] Nicotinic acid (3.604 g, 29.25 mmol) was dissolved in 100 mL
of deionized (DI) water by heating to 80-85.degree. C. Cysteine
HCl.H.sub.2O (2.65 g, 15.0 mmol) was added as a solid, followed by
a 10 mL addition of DI water. The solution was stirred for 5 min,
until all the solid had dissolved. CrCl.sub.3.6H.sub.2O (4.06 g,
15.3 mmol) was slowly added as a solid, followed by a 10 mL rinse
of DI water. The solution immediately turned a dark blue-green. The
solution was stirred at 75-80.degree. C. for 8 hours. The heat was
removed and the reaction was stirred overnight. The cooled solution
was frozen and lyophilized to afford 9.2 g of a dark green
solid.
Evaluation of Chromium (III) Complexes as Disclosed Herein
[0068] Chromium complexes were evaluated for bioavailability and in
vivo activity. The compounds were tested in comparison with
chromium compounds that are known in the art, including, for
example, chromium picolinate and chromium nicotinate.
[0069] In Vivo Assessment: Obese Zucker rats were used as an ideal
type II diabetic animal model for this evaluation. The rats,
obtained at five weeks of age and acclimated for two days, were
fasted overnight. The rats were then weighed and had blood
collected to obtain a baseline reading. The rats were divided into
groups to test each compound and fed a Purina.RTM. 5008 lab chow
diet (Wilmington, Mass.) for eight weeks.
[0070] The compounds tested are listed in Table I below.
Chromium-containing compounds were administered daily at 400 .mu.g
elemental chromium (III) per kilogram body weight for eight weeks.
Niacin, arginine, and cysteine were tested at the amounts
equivalent to the individual compound present in the complexes
containing 400 .mu.g elemental chromium (III) and niacin, arginine,
and cysteine, respectively. Niacin was administered daily at
approximately 1.9 mg per kilogram body weight. Arginine and
cysteine were each administered daily at approximately 1 mg per
kilogram body weight. The control group received saline solution
alone daily.
[0071] The study monitored the following parameters: (1) body
weight; (2) food intake; (3) vitamin C levels; (4) fasting glucose
levels; (5) lipid profile (total cholesterol (TC), high density
lipoprotein (HDL), and triglycerides (TG)); (6) glycosylated
hemoglobin (HbAlc); (7) red blood cell lipid peroxidase (RBC lipid
peroxidase); (8) adiponectin; (9) plasma C-reactive protein (CRP);
(10) monocyte chemotactic protein (MCP-1); (11) intracellular
adhesion molecule (ICAM); (12) glutathione (GSH); (13) retinol
binding protein (RBP-4); (14) leptin; (15) cyclic adenosine
monophosphate (cAMP); (16) tumor necrosis factor (TNF.alpha.); (17)
interleukin 6 (IL-6); and (18) oxidative DNA damage (OxyDNA).
[0072] Food intake was measured at 5 and 7 weeks. At the end of
eight weeks, the rates were fasted overnight and then euthanized.
Blood was collected, and the plasma was isolated by centrifugation.
CRP, MCP-1, ICAM, and adiponectin levels in the plasma were
determined by sandwich ELISA method using commercially available
kits from Fisher Thermo Scientific Co. (Rockford, Ill.). Oxidative
stress was determined by measuring malondialdehyde (an end product
of lipid peroxidation) by its reaction with thiobarbituric acid.
Vitamin C concentration in the plasma was also determined.
[0073] Glycosylated hemoglobin was determined using Glyco-Tek
Affinity column kits and reagents available commercially from
Helena Laboratories (Beaumont, Tex.). Glucose levels were
determined using glucose oxidase measured with an Accu-Chek.RTM.
Advantage glucometer available commercially from Boehringer Manheim
Corp. (Indianapolis, Ind.).
[0074] The following data in Tables II-V demonstrates the
beneficial effects of chromium dinicocysteinate, for example, by
reducing fasting glucose, glycosylated hemoglobin, C-reactive
protein, monocyte chemotactic protein-1, intracellular adhesion
molecule, and red blood cell lipid peroxidation. Blood vitamin C
levels and adiponectin levels were increased with the
administration of chromium dinicocysteinate.
[0075] In addition, chromium dinicocysteinate outperformed other
chromium compounds known for chromium supplementation, such as, for
example, chromium nicotinate and chromium picolinate. Specifically,
over chromium nicotinate, chromium picolinate, and chromium
histidinate, chromium dinicocysteinate demonstrated decreased
levels of fasting glucose, ICAM, C-reactive protein, and
glycosylated hemoglobin and increased levels of vitamin C and
adiponectin. It also demonstrated lower levels of MCP-1 and red
blood cell lipid peroxidase over chromium picolinate. Chromium
dinicocysteinate lowered levels of TNF.alpha. over chromium
picolinate and chromium histidinate. Finally, chromium
dinicocysteinate demonstrated reduced food intake, decreased levels
of fasting glucose, glycosylated hemoglobin, red blood cell lipid
peroxidation, TNF.alpha., OxyDNA, AST and ALT, and increased levels
of vitamin C and adiponectin as compared to the chromium
nicotinate-glycine-cysteine-glutamic acid GTF complex proposed by
Toepfer et al.
[0076] The following legend describes each abbreviation used in the
following tables for each compound.
TABLE-US-00001 TABLE I Abbreviations of the compounds tested
Abbreviation Identification BL Baseline Control Control CrN
Chromium Nicotinate CrP Chromium Picolinate CrP + Biotin Chromium
Picolinate + Biotin Cr(nic).sub.2Trp Chromium Dinicotinate
Tryptophanate Cr(nic).sub.2Phe Chromium Dinicotinate Phenylalanine
Cr(nic).sub.2Asp Chromium Dinicotinate Aspartate Cr(nic).sub.2Glu
Chromium Dinicotiante Glutamate Cr(nic).sub.2Cys Chromium
Dinicocysteinate Cr(nic).sub.2Gly Chromium Dinicotinate Glycinate
Niacin Niacin Cr(nic).sub.2Arg Chromium Dinicotinate Argininate Arg
Arginine Cys Cysteine CrNicGlyCysGlu Compound according to Toepfer
et al..sup.1 including chromium, nicotinic acid, glycine, cysteine,
and glutamic acid CrHis Chromium Histidinate CrP Chromium
Propionate Cr454 Chromium 454 .TM. is created by the reaction of
trivalent chromium with high-activity, water soluble small
molecules from a GRAS Brewer's yeast extract CrTrp Chromium
Tryptophanate CrCin Chromium Cinnamate Cr(5HTP).sub.3 Chromium
tris(5-hydroxytryptophan) Cr(nic).sub.2Gluc Chromium
Glucodinicotinate Cr(Phe).sub.3 Chromium Triphenylalanine
.sup.1Toepfer et al., Preparation of Chromium-Containing Material
of Glucose Tolerance Factor Activity from Brewer's Yeast Extracts
and by Synthesis, J. Agric. Food Chem., 25(1): 162-166 (1977).
TABLE-US-00002 TABLE II Effect on Blood Lipid Profiles Body Food
Food Weight at Intake Intake Total Total Sacrificing 5 wks 7 wks
Cholesterol Triglyceride HDL Chol/HDL N (g) (g/day) (g/day) (mg/dL)
(mg/dL) (mg/dL) (ratio) Baseline 27 151.72 .+-. -- -- 128.91 .+-.
181.26 .+-. 47.89 .+-. 2.78 .+-. 5.21 3.25 15.97 1.30 0.05 Control
25 358.73 .+-. 38.55 .+-. 37.78 .+-. 189.31 .+-. 518.50 .+-. 69.58
.+-. 2.84 .+-. 4.98 1.25 1.33 4.13 24.96 1.43 0.05 CrN 20 366.33
.+-. 36.52 .+-. 39.6 .+-. 184.27 .+-. 514.49 .+-. 67.54 .+-. 2.83
.+-. 5.88 1.22 1.46 3.94 29.51 1.47 0.05 CrP 19 368.67 .+-. 35.85
.+-. 40.79 .+-. 191.80 .+-. 625.35 .+-. 66.01 .+-. 3.02 .+-. 5.99
1.05 1.61 6.68 82.99 1.80 0.09 Crp + Biotin 7 364.57 .+-. 38.00
.+-. 40.53 .+-. 187.86 .+-. 478.71 .+-. 67.14 .+-. 2.80 .+-. 9.65
2.09 1.76 3.65 57.55 1.47 0.07 Cr(nic).sub.2Trp 7 368.57 .+-. 40.49
.+-. 40.78 .+-. 183.17 .+-. 470.67 .+-. 69.43 .+-. 2.86 .+-. 11.54
1.97 1.29 9.51 71.53 3.20 0.15 Cr(nic).sub.2Phe 7 358.00 .+-. 38.16
.+-. 39.22 .+-. 201.71 .+-. 583.00 .+-. 70.43 .+-. 2.87 .+-. 5.52
1.17 1.51 7.74 60.36 2.62 0.08 Cr(nic).sub.2Asp 6 375.67 .+-. 38.90
.+-. 40.00 .+-. 202.17 .+-. 699.67 .+-. 70.17 .+-. 2.88 .+-. 7.38
1.58 0.53 4.21 75.01 1.08 0.07 Cr(nic).sub.2Glu 7 392.86 .+-. 37.51
.+-. 39.92 .+-. 192.14 .+-. 666.50 .+-. 65.57 .+-. 2.94 .+-. 15.58
2.04 2.17 9.90 89.36 3.22 0.09 Cr(nic).sub.2Cys 13 375.08 .+-.
34.43 .+-. 38.62 .+-. 188.46 .+-. 550.89 .+-. 66.46 .+-. 3.00 .+-.
7.83 1.10 1.62 5.00 47.96 1.42 0.08 Cr(nic).sub.2Gly 7 371.43 .+-.
34.29 .+-. 37.14 .+-. 193.14 .+-. 610.33 .+-. 64.57 .+-. 3.00 .+-.
9.54 1.77 1.54 9.84 75.44 2.58 0.13 Niacin 7 368.33 .+-. 37.92 .+-.
39.67 .+-. 192.17 .+-. 486.67 .+-. 70.33 .+-. 2.73 .+-. 9.90 1.60
2.96 8.97 41.09 2.44 0.06 Cr(nic).sub.2Arg 6 367.33 .+-. 40.22 .+-.
45.28 .+-. 186.16 .+-. 554.65 .+-. 69.78 .+-. 3.00 .+-. 7.07 1.85
3.15 2.94 26.27 1.84 0.04 Arg 6 378.00 .+-. 37.73 .+-. 46.80 .+-.
173.96 .+-. 485.74 .+-. 68.69 .+-. 2.86 .+-. 12.07 1.01 3.35 7.99
24.07 3.69 0.11 Cys 6 364.80 .+-. 39.40 .+-. 39.87 .+-. 183.58 .+-.
503.57 .+-. 68.26 .+-. 3.02 .+-. 13.98 2.24 1.23 8.940 40.47 3.31
0.07 CrNicGlyCysGlu 6 357.33 .+-. 41.61 .+-. 42.78 .+-. 168.08 .+-.
431.27 .+-. 69.08 .+-. 2.73 .+-. 4.31 2.93 1.97 7.29 40.95 2.72
0.04 CrHis 6 367.00 .+-. -- -- 189.62 .+-. 503.93 .+-. 66.90 .+-.
2.86 .+-. 9.15 7.86 46.27 2.56 0.05 CrProp 6 338.40 .+-. -- --
170.56 .+-. 456.14 .+-. 64.19 .+-. 2.68 .+-. 9.39 6.82 52.20 1.61
0.05 Cr454 6 352.00 .+-. -- -- 168.08 .+-. 507.57 .+-. 60.95 .+-.
2.78 .+-. 6.02 10.86 26.83 3.14 0.05 CrTrp 6 417.71 .+-. -- --
208.09 .+-. 921.22 .+-. 60.54 .+-. 3.46 .+-. 10.95 13.52 147.69
1.51 0.18 CrCin 7 374.00 .+-. -- -- 191.11 .+-. 507.35 .+-. 65.44
.+-. 2.94 .+-. 5.89 8.68 47.57 2.10 0.0548 Cr(5HTP).sub.3 7 343.43
.+-. -- -- 184.77 .+-. 452.96 .+-. 68.84 .+-. 2.72 .+-. 10.10 5.32
51.83 2.19 0.07 Cr(nic).sub.2Gluc 7 387.67 .+-. -- -- 197.33 .+-.
534.43 .+-. 65.71 .+-. 3.06 .+-. 12.06 6.96 46.77 3.55 0.12
Cr(Phe).sub.3 7 382.57 .+-. -- -- 169.68 .+-. 467.04 .+-. 60.82
.+-. 2.84 .+-. 17.18 9.58 43.70 3.91 0.12 Each value represents
mean .+-. SE
TABLE-US-00003 TABLE III Effect on Key Biological Parameters RBC
Fasting Vitamin Lipid Glucose HbA1c C GSH Peroxidase N (mg/dL) (%)
(mg %) (umol/g) (nmol/mL) Baseline 27 233.74 .+-. 5.61 .+-. 0.27
.+-. 4.21 .+-. 1.35 .+-. 8374 0.12 0.01 0.11 0.28 Control 25 606.25
.+-. 16.59 .+-. 0.15 .+-. 4.85 .+-. 2.26 .+-. 21.27 0.19 0.01 1.12
0.08 CrN 20 577.29 .+-. 15.80 .+-. 0.16 .+-. 4.76 .+-. 1.94 .+-.
21.44 0.24 0.01 0.17 0.12 CrP 19 527.28 .+-. 15.54 .+-. 0.15 .+-.
4.84 .+-. 2.05 .+-. 23.95 0.30 0.01 0.10 0.12 CrP + Biotin 7 527.14
.+-. 15.79 .+-. 0.12 .+-. 5.25 .+-. 2.19 .+-. 15.08 0.44 0.01 0.14
0.20 Cr(nic).sub.2Trp 7 513.43 .+-. 15.55 .+-. 0.13 .+-. 5.35 .+-.
1.96 .+-. 24.15 0.30 0.01 0.14 0.24 Cr(nic).sub.2Phe 7 510.43 .+-.
15.95 .+-. 0.15 .+-. 5.05 .+-. 1.97 .+-. 25.45 0.24 0.01 0.17 0.14
Cr(nic).sub.2Asp 6 496.33 .+-. 15.62 .+-. 0.12 .+-. 5.15 .+-. 2.02
.+-. 29.35 0.35 0.01 0.15 0.22 Cr(nic).sub.2Glu 7 490.43 .+-. 14.69
.+-. 0.14 .+-. 5.05 .+-. 2.30 .+-. 40.69 0.57 0.01 0.26 0.21
Cr(nic).sub.2Cys 13 446.24 .+-. 14.76 .+-. 0.18 .+-. 4.74 .+-. 1.81
.+-. 31.04 0.51 0.01 0.17 0.12 Cr(nic).sub.2Gly 7 448.29 .+-. 14.45
.+-. 0.15 .+-. 4.86 .+-. 2.11 .+-. 37.83 0.67 0.01 0.19 0.19 Niacin
7 543.00 .+-. 15.60 .+-. 0.15 .+-. 5.32 .+-. 1.82 .+-. 20.99 0.47
0.01 0.16 0.16 Cr(nic).sub.2Arg 6 628.87 .+-. 16.56 .+-. 0.14 .+-.
4.89 .+-. 2.47 .+-. 41.70 0.22 0.01 0.30 0.11 Arg 6 446.60 .+-.
15.88 .+-. 0.14 .+-. 4.77 .+-. 2.23 .+-. 35.43 0.43 0.01 0.15 0.04
Cys 6 483.49 .+-. 15.17 .+-. 0.13 .+-. 4.70 .+-. 2.34 .+-. 38.22
0.21 0.01 0.26 0.18 CrNicGlyCysGlu 6 533.63 .+-. 15.90 .+-. 0.14
.+-. 4.72 .+-. 2.34 .+-. 54.48 0.32 0.01 0.15 0.05 CrHis 6 592.01
.+-. 15.89 .+-. -- 4.68 .+-. 1.91 .+-. 34.43 0.37 0.28 0.07 CrProp
6 549.49 .+-. 16.90 .+-. -- 4.76 .+-. 2.18 .+-. 49.44 0.16 0.20
0.28 Cr454 6 573.60 .+-. 16.47 .+-. -- 4.44 .+-. 2.39 .+-. 24.04
0.55 0.46 0.45 CrTrp 7 402.77 .+-. 14.16 .+-. -- 4.54 .+-. 1.78
.+-. 28.41 0.28 0.24 0.12 CrCin 7 557.37 .+-. 15.61 .+-. -- 5.00
.+-. 3.12 .+-. 36.91 0.33 0.12 0.44 Cr(5HTP).sub.3 7 585.25 .+-.
16.15 .+-. -- 5.04 .+-. 1.93 .+-. 31.23 0.30 0.08 0.08
Cr(nic).sub.2Gluc 7 474.38 .+-. 15.21 .+-. -- 4.62 .+-. 2.00 .+-.
60.73 0.59 0.17 0.12 Cr(Phe).sub.3 7 549.70 .+-. 15.04 .+-. -- 4.50
.+-. 2.01 .+-. 45.16 0.65 0.35 0.01 Each value represents the mean
.+-. SE.
TABLE-US-00004 TABLE IV Effect on Vascular Inflammation Markers
MCP-1 CRP RBP-4 Leptin ICAM Adiponectin N (pg/mL) (ng/mL) (ng/mL)
(pg/mL) (pg/mL) (ng/mL) Baseline 27 2141.63 .+-. 445.91 .+-.
46064.49 .+-. 329.56 .+-. 14804.29 .+-. 38183.23 .+-. 148.03 10.65
2301.82 13.92 466.52 1392.69 Control 25 4080.10 .+-. 712.99 .+-.
49618.54 .+-. 412.78 .+-. 18212.60 .+-. 11399.80 .+-. 367.64 20.46
1914.00 22.70 530.12 288.08 CrN 20 3175.02 .+-. 689.86 .+-.
48498.10 .+-. 374.99 .+-. 18029.45 .+-. 10575.95 .+-. 458.52 17.56
1656.82 23.90 333.83 355.40 CrP 19 4549.99 .+-. 654.78 .+-.
49236.91 .+-. 399.55 .+-. 20428.47 .+-. 11229.84 .+-. 541.23 27.22
42144.02 22.51 1568.35 564.20 CrP + Biotin 7 2597.63 .+-. 636.65
.+-. 53184.29 .+-. 284.1 7.+-. 17555.81 .+-. 8898.57 .+-. 630.54
27.73 2391.91 13.21 484.45 605.35 Cr(nic).sub.2Trp 7 2607.07 .+-.
617.66 .+-. 60017.14 .+-. 363.21 .+-. 17950.97 .+-. 11752.86 .+-.
635.54 19.13 2158.23 17.57 673.35 1114.20 Cr(nic).sub.2Phe 7
1755.25 .+-. 633.55 .+-. 53891.43 .+-. 351.16 .+-. 17480.50 .+-.
12712.86 .+-. 348.03 24.11 3138.66 11.94 692.46 506.92
Cr(nic).sub.2Asp 6 1431.68 .+-. 602.51 .+-. 57870.00 .+-. 406.15
.+-. 17985.91 .+-. 12078.33 .+-. 351.70 35.00 7839.39 37.40 678.21
755.93 Cr(nic).sub.2Glu 7 3197.58 .+-. 519.94 .+-. 46720.00 .+-.
378.02 .+-. 16980.59 .+-. 12446.67 .+-. 684.32 21.48 2053.76 20.80
375.03 1420.72 Cr(nic).sub.2Cys 13 2829.17 .+-. 569.96 .+-.
49670.13 .+-. 407.32 .+-. 17057.51 .+-. 12732.11 .+-. 505.01 24.87
2905.84 22.68 407.77 981.42 Cr(nic).sub.2Gly 7 2897.61 .+-. 535.98
.+-. 54900.00 .+-. 461.16 .+-. 16691.40 .+-. 12552.86 .+-. 953.13
21.20 3370.63 63.69 361.13 961.72 Niacin 7 1365.66 .+-. 548.27 .+-.
53945.00 .+-. 363.96 .+-. 16855.83 .+-. 8872.00 .+-. 600.75 23.65
3658.98 15.37 386.02 479.48 Cr(nic).sub.2Arg 6 3235.62 .+-. 628.49
.+-. 52537.50 .+-. 335.80 .+-. 18591.76 .+-. 9279.38 .+-. 255.96
14.76 891.06 15.48 852.37 481.63 Arg 6 3878.85 .+-. 617.34 .+-.
47211.67 .+-. 431.72 .+-. 17940.91 .+-. 9958.63 .+-. 417.45 29.21
2102.85 46.39 474.12 176.77 Cys 6 2769.76 .+-. 551.39 .+-. 50915.00
.+-. 477.65 .+-. 18512.88 .+-. 11532.37 .+-. 222.75 38.24 3416.90
49.02 670.51 805.94 CrNicGlyCysGlu 6 2911.81 .+-. 564.10 .+-.
46577.78 .+-. 361.11 .+-. 18621.43 .+-. 10152.28 .+-. 174.07 14.54
2215.22 29.13 632.75 346.38 CrHis 6 2974.79 .+-. 710.26 .+-.
51112.60 .+-. 436.96 .+-. -- 11455.87 .+-. 862.09 133.82 2856.20
45.86 731.12 CrProp 6 4497.83 .+-. 820.04 .+-. 49851.91 .+-. 393.47
.+-. -- 12903.52 .+-. 547.42 81.86 3664.21 33.43 781.29 Cr454 6
3232.18 .+-. 747.02 .+-. 48791.98 .+-. 364.80 .+-. -- 11055.14 .+-.
477.26 60.39 2827.93 54.52 201.42 CrTrp 7 2578.59 .+-. 609.82 .+-.
60593.24 .+-. 601.72 .+-. -- 11122.93 .+-. 388.46 43.37 2972.93
77.55 345.96 CrCin 7 2428.69 .+-. 587.35 .+-. 47905.13 .+-. 379.18
.+-. -- 11507.60 .+-. 329.03 21.41 1751.73 31.66 563.90
Cr(5HTP).sub.3 7 1932.18 .+-. 589.26 .+-. 44517.99 .+-. 359.61 .+-.
-- 11831.51 .+-. 280.81 24.48 1550.87 27.52 244.04
Cr(nic).sub.2Gluc 7 1977.46 .+-. 530.74 .+-. 48851.14 .+-. 388.37
.+-. -- 11448.39 .+-. 211.78 22.37 2756.82 56.16 412.77
Cr(Phe).sub.3 7 2064.89 .+-. 533.12 .+-. 48555.98 .+-. 387.92 .+-.
-- 11243.47 .+-. 211.78 22.37 2756.82 56.16 412.77 Each value
represents the mean .+-. SE.
TABLE-US-00005 TABLE V Effect on Other Vascular Inflammation
Markers cAMP TNF.alpha. IL-6 OxyDNA N (pmol/mL) (pg/mL) (pg/mL)
(ng/mL) Baseline 27 43.08 .+-. 3.26 26.53 .+-. 4.18 96.34 .+-. 1.57
55.96 .+-. 3.02 Control 25 30.21 .+-. 1.15 42.88 .+-. 10.05 91.59
.+-. 1.75 49.10 .+-. 1.52 CrN 20 32.29 .+-. 3.59 17.02 .+-. 3.59
91.42 .+-. 2.40 49.03 .+-. 1.56 CrP 19 29.17 .+-. 2.00 33.66 .+-.
5.27 94.13 .+-. 2.47 49.02 .+-. 2.31 CrP + Biotin 7 30.47 .+-. 2.41
-- 91.05 .+-. 1.81 40.30 .+-. 2.35 Cr(nic).sub.2Trp 7 30.36 .+-.
1.48 -- 92.09 .+-. 1.89 39.35 .+-. 3.48 Cr(nic).sub.2Phe 7 31.60
.+-. 1.22 -- 84.82 .+-. 1.60 40.63 .+-. 3.74 Cr(nic).sub.2Asp 6
27.95 .+-. 1.22 -- 88.91 .+-. 2.25 41.92 .+-. 1.46 Cr(nic).sub.2Glu
7 29.97 .+-. 1.43 -- 89.50 .+-. 3.22 43.62 .+-. 2.69
Cr(nic).sub.2Cys 13 33.26 .+-. 2.61 17.72 .+-. 3.35 95.02 .+-. 1.99
46.92 .+-. 2.24 Cr(nic).sub.2Gly 7 31.40 .+-. 1.57 -- 91.96 .+-.
3.83 45.62 .+-. 1.86 Niacin 7 34.27 .+-. 1.47 -- 93.76 .+-. 1.60
47.31 .+-. 2.78 Cr(nic).sub.2Arg 6 38.96 .+-. 3.88 17.84 .+-. 1.26
92.35 .+-. 2.79 58.26 .+-. 2.57 Arg 6 34.98 .+-. 0.92 25.45 .+-.
0.93 97.80 .+-. 0.93 62.90 .+-. 2.71 Cys 6 32.94 .+-. 1.79 24.28
.+-. 3.84 95.33 .+-. 2.33 54.56 .+-. 5.69 CrNicGlyCysGlu 6 35.55
.+-. 0.89 23.37 .+-. 9.31 97.80 .+-. 1.42 56.07 .+-. 6.20 CrHis 6
28.07 .+-. 2.83 24.72 .+-. 5.24 91.98 .+-. 8.19 57.72 .+-. 2.86
CrProp 6 26.95 .+-. 1.13 41.14 .+-. 10.06 90.50 .+-. 14.02 47.19
.+-. 5.07 Cr454 6 26.41 .+-. 3.20 20.69 .+-. 6.10 82.95 .+-. 5.25
46.57 .+-. 4.80 CrTrp 7 20.03 .+-. 0.96 36.61 .+-. 14.89 86.62 .+-.
6.81 45.14 .+-. 2.77 CrCin 7 29.24 .+-. 0.61 8.62 .+-. 4.90 92.94
.+-. 13.21 48.71 .+-. 1.25 Cr(5HTP).sub.3 7 25.64 .+-. 2.39 13.35
.+-. 8.28 94.32 .+-. 11.71 45.89 .+-. 4.53 Cr(nic).sub.2Gluc 7
22.25 .+-. 1.58 29.40 .+-. 8.87 81.59 .+-. 5.21 45.10 .+-. 3.76
Cr(Phe).sub.3 7 29.62 .+-. 1.01 15.92 .+-. 7.76 89.82 .+-. 8.55
53.11 .+-. 4.76 Each value represents the mean .+-. SE.
TABLE-US-00006 TABLE VI Effect on Clinical Chemistry Abbreviations:
AP: Alkaline phosphatase; BUN: Blood Urea Nitrogen; CRT:
Creatinine; AST: Aspartate Aminotransferase; ALT: Alanine
Aminotransferease. Total Anion BUN/CRT AP BUN CRT AST ALT Bilirubin
Gap Ratio N (u/L) (mg/dL) (mg/dL) (u/L) (u/L) (mg/dL) (.mu./L)
(mMol/L) Baseline 27 41.36 .+-. 14.74 .+-. 0.35 .+-. 271.83 .+-.
72.13 .+-. 0.07 .+-. 37.22 .+-. 43.17 .+-. 6.01 0.39 0.01 33.85
1.72 0.00 0.91 1.49 Control 25 55.86 .+-. 20.27 .+-. 0.43 .+-.
375.15 .+-. 244.09 .+-. 0.07 .+-. 28.77 .+-. 48.18 .+-. 6.61 0.64
0.00 90.35 62.90 0.01 0.62 1.93 CrN 20 45.39 .+-. 20.58 .+-. 0.43
.+-. 227.67 .+-. 133.53 .+-. 0.07 .+-. 28.00 .+-. 49.53 .+-. 5.47
1.10 0.01 33.31 11.91 0.01 0.53 2.96 CrP 19 40.06 .+-. 20.22 .+-.
0.41 .+-. 362.44 .+-. 179.78 .+-. 0.07 .+-. 29.61 .+-. 49.72 .+-.
4.72 1.09 0.01 51.87 26.86 0.01 0.78 2.57 CrP + Biotin 7 64.00 .+-.
19.14 .+-. 0.41 .+-. 160.29 .+-. 100.71 .+-. 0.10 .+-. 25.86 .+-.
46.50 .+-. 5.78 0.86 0.01 22.59 10.08 0.00 1.16 2.57
Cr(nic).sub.2Trp 7 61.43 .+-. 19.57 .+-. 0.40 .+-. 149.71 .+-.
89.29 .+-. 0.09 .+-. 27.14 .+-. 48.93 .+-. 10.71 1.04 0.00 28.07
7.69 0.01 1.37 2.61 Cr(nic).sub.2Phe 7 51.43 .+-. 19.71 .+-. 0.39
.+-. 135.29 .+-. 108.00 .+-. 0.10 .+-. 27.71 .+-. 51.43 .+-. 8.25
0.57 0.01 16.31 10.52 0.00 0.78 1.88 Cr(nic).sub.2Asp 6 53.00 .+-.
18.00 .+-. 0.42 .+-. 120.33 .+-. 96.00 .+-. 0.10 .+-. 26.00 .+-.
45.83 .+-. 2.35 0.84 0.02 17.89 6.14 0.00 0.77 1.90 Cr(nic)2Glu 7
52.86 .+-. 18.43 .+-. 0.41 .+-. 176.71 .+-. 105.57 .+-. 0.10 .+-.
27.00 .+-. 44.71 .+-. 9.01 0.69 0.01 23.22 12.83 0.00 1.38 2.03
Cr(nic).sub.2Cys 13 35.15 .+-. 18.23 .+-. 0.41 .+-. 261.17 .+-.
99.00 .+-. 0.05 .+-. 29.62 .+-. 44.92 .+-. 3.95 0.64 0.00 41.43
6.03 0.01 0.58 1.70 Cr(nic).sub.2Gly 7 40.50 .+-. 16.71 .+-. 0.40
.+-. 176.71 .+-. 90.83 .+-. 0.10 .+-. 29.43 .+-. 41.07 .+-. 3.53
0.47 0.00 23.22 3.46 0.00 1.04 1.53 Niacin 7 46.50 .+-. 19.33 .+-.
0.40 .+-. 261.17 .+-. 126.17 .+-. 0.10 .+-. 28.67 .+-. 48.33 .+-.
8.01 0.84 0.00 41.43 15.74 0.00 0.84 2.11 Cr(nic).sub.2Arg 6 40.20
.+-. 20.00 .+-. 0.42 .+-. 241.50 .+-. 147.33 .+-. 0.00 .+-. 27.60
.+-. 48.33 .+-. 5.96 0.68 0.02 29.34 24.54 0.00 0.51 2.09 Arg 6
35.50 .+-. 21.00 .+-. 0.40 .+-. 276.80 .+-. 179.20 .+-. 0.00 .+-.
27.75 .+-. 52.60 .+-. 7.98 0.45 0.00 48.90 23.91 0.00 0.85 1.12 Cys
6 36.40 .+-. 18.50 .+-. 0.42 .+-. 218.20 .+-. 119.20 .+-. 0.00 .+-.
29.20 .+-. 43.00 .+-. 4.65 0.29 0.02 25.46 11.37 0.00 0.49 2.53
CrNicGlyCysGlu 6 36.16 .+-. 19.33 .+-. 0.40 .+-. 241.17 .+-. 160.00
.+-. 0.00 .+-. 28.33 .+-. 48.67 .+-. 8.33 1.05 0.00 50.71 39.72
0.00 0.67 2.55 CrHis 6 35.25 .+-. 19.50 .+-. 0.40 .+-. 248.00 .+-.
168.25 .+-. 0.10 .+-. 29.75 .+-. 48.75 .+-. 11.52 0.50 0.00 19.22
22.41 0.00 1.32 1.25 CrProp 6 32.20 .+-. 22.25 .+-. 0.40 .+-.
347.25 .+-. 207.00 .+-. 0.10 .+-. 29.20 .+-. 53.40 .+-. 10.68 0.48
0.00 50.92 49.00 0.00 0.97 2.77 Cr454 6 31.50 .+-. 20.00 .+-. 0.38
.+-. 439.50 .+-. 213.67 .+-. 0.10 .+-. 30.00 .+-. 48.80 .+-. 10.38
1.15 0.02 90.07 51.00 0.00 1.55 3.1209 CrTrp 7 30.00 .+-. 17.43
.+-. 0.39 .+-. 237.43 .+-. 124.29 .+-. 0.10 .+-. 30.14 .+-. 45.86
.+-. 7.79 0.61 0.01 55.49 16.77 0.00 0.34 2.82 CrCin 7 30.71 .+-.
21.43 .+-. 0.41 .+-. 230.14 .+-. 158.71 .+-. 0.10 .+-. 29.86 .+-.
51.86 .+-. 6.82 1.38 0.01 20.78 23.76 0.00 0.26 1.72 Cr(5HTP).sub.3
7 25.00 .+-. 22.86 .+-. 0.40 .+-. 267.43 .+-. 167.00 .+-. 0.10 .+-.
29.43 .+-. 57.29 .+-. 8.28 1.39 0.00 22.84 25.31 0.00 0.53 3.42
Cr(nic).sub.2Gluc 7 27.00 .+-. 21.33 .+-. 0.40 .+-. 238.67 .+-.
138.00 .+-. 0.10 .+-. 29.00 .+-. 53.67 .+-. 8.53 0.84 0.00 25.03
.+-. 12.83 0.00 1.03 2.12 Cr(Phe)3 7 30.00 .+-. 20.71 .+-. 0.40
.+-. 276.43 .+-. 186.29 .+-. 0.10 .+-. 29.00 .+-. 52.14 .+-. 7.65
1.08 0.00 48.30 .+-. 39.13 0.00 0.52 2.73 Each value represents the
mean .+-. SE
[0077] The foregoing description is given for clearness of
understanding only, and no unnecessary limitations should be
understood therefrom, as modifications within the scope of the
invention may be apparent to those having ordinary skill in the
art.
* * * * *