U.S. patent application number 16/715905 was filed with the patent office on 2020-09-24 for compositions of citrulline.
The applicant listed for this patent is Hubei Grand Biotechnology Co. Ltd.. Invention is credited to Hui Ouyang.
Application Number | 20200297677 16/715905 |
Document ID | / |
Family ID | 1000004537108 |
Filed Date | 2020-09-24 |
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United States Patent
Application |
20200297677 |
Kind Code |
A1 |
Ouyang; Hui |
September 24, 2020 |
COMPOSITIONS OF CITRULLINE
Abstract
The present disclosure relates generally to supplement
compositions containing citrulline and taurine at a weight ratio of
4.56:1 to 2.57:1 which have been demonstrated to have a pleasant
flavor and can enhance the exercise endurance in individuals in a
synergistic manner.
Inventors: |
Ouyang; Hui; (Hubei,
CN) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Hubei Grand Biotechnology Co. Ltd. |
Huangshi |
|
CN |
|
|
Family ID: |
1000004537108 |
Appl. No.: |
16/715905 |
Filed: |
December 16, 2019 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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16374625 |
Apr 3, 2019 |
10548865 |
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16715905 |
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PCT/CN2019/078623 |
Mar 19, 2019 |
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16374625 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 31/185 20130101;
A61K 31/198 20130101; A23L 2/52 20130101; A61K 9/0095 20130101;
A61P 21/00 20180101; A23L 33/16 20160801; A23V 2002/00 20130101;
A61P 21/06 20180101; A23L 33/175 20160801 |
International
Class: |
A61K 31/198 20060101
A61K031/198; A61P 21/00 20060101 A61P021/00; A61P 21/06 20060101
A61P021/06; A23L 33/175 20060101 A23L033/175; A61K 9/00 20060101
A61K009/00; A23L 33/16 20060101 A23L033/16; A61K 31/185 20060101
A61K031/185; A23L 2/52 20060101 A23L002/52 |
Claims
1. A solid composition comprising citrulline and taurine, wherein
the citrulline and the taurine have a weight ratio of 5:1 to 1:5
and together constitute at least 55% w/w of the solid
composition.
2. The solid composition of claim 1, wherein the citrulline and the
taurine have a weight ratio of 5:1 to 1:1.
3. The solid composition of claim 2, which does not include more
than 20% w/w of the combination of any other amino acid, vitamins,
peptides, proteins, herb extracts, fatty acids, fibers, probiotics,
glucosamine, chondroitin, and CoQ10.
4. The solid composition of claim 3, wherein the citrulline and the
taurine have a weight ratio of 4:1 to 2.85:1.
5. The solid composition of claim 3, wherein the citrulline and the
taurine have a weight ratio of 3.55:1 to 3:1.
6. The solid composition of claim 3, wherein the citrulline and the
taurine have a weight ratio of 3.35:1 to 3.08:1.
7. The solid composition of claim 3, wherein the citrulline and the
taurine together constitute at least 60% w/w of the solid
composition.
8. The solid composition of claim 3, wherein the citrulline and the
taurine together constitute at least 75% w/w of the solid
composition.
9. The solid composition of claim 1, which is provided in the form
of powders, tablets, granules, or capsules.
10. The solid composition of claim 9, wherein the tablets or
granules are effervescent tablets or granules.
11. A drink comprising citrulline and taurine, wherein the
citrulline and the taurine have a weight ratio of 5:1 to 1:5 and
together constitute at least 55% w/w of total solid ingredients
dissolved in the drink.
12. The drink of claim 11, wherein the citrulline and the taurine
have a weight ratio of 5:1 to 1:1.
13. The drink of claim 11, wherein the citrulline and the taurine
have a weight ratio of 4:1 to 2.85:1.
14. The drink of claim 11, wherein the citrulline and the taurine
have a weight ratio of 3.55:1 to 3:1.
15. The drink of claim 11, wherein the citrulline and the taurine
have a weight ratio of 3.35:1 to 3.08:1.
16. The drink of claim 12, wherein the citrulline and the taurine
together constitute at least 60% w/w of the total solid ingredients
dissolved in the drink.
17. The drink of claim 12, wherein the citrulline and the taurine
together constitute at least 75% w/w of the total solid ingredients
dissolved in the drink.
18. A method for treating hypertension or prehypertension in a
mammalian subject, comprising orally administering to the subject
the drink of claim 11.
19. The method of claim 18, wherein about 0.1 g to about 100 g of
the combination of citrulline and the taurine are administered
daily.
20. The method of claim 18, wherein about 1 g to 10 g of the
combination of citrulline and the taurine are administered daily.
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] The present application is a continuation application of
U.S. application Ser. No. 16/374,625, filed Apr. 3, 2019, which is
a continuation application of International Application
PCT/CN2019/078623, filed Mar. 19, 2019, the contents of each of
which are incorporated by reference in their entirety into the
present disclosure.
BACKGROUND
[0002] L-citrulline, or simply citrulline, is an .alpha.-amino
acid, with its name derived from citrullus, the Latin word for
watermelon. Citrulline is a key intermediate in the urea cycle, the
pathway by which mammals excrete ammonia by converting it into
urea. Citrulline is also produced as a byproduct of the enzymatic
production of nitric oxide from the amino acid arginine, catalyzed
by nitric oxide synthase.
[0003] Citrulline boosts nitric oxide (NO) production in the body.
Nitric oxide helps arteries relax and work better, which improves
blood flow throughout the body. NO is an integral part of the
inflammatory phase, functioning as a regulatory mechanism to
mediate epithelialization, angiogenesis, and collagen deposition
crucial to the proliferative phase. NO-induced vasodilatation acted
as a host-protective agent by killing pathogens and by increasing
blood flow to wounds. The supplement of citrulline is used to lower
blood pressure in people with prehypertension. Citrulline
supplements may also ease symptoms of mild-to-moderate erectile
dysfunction (ED) and help to address blood vessel problems such as
slow wound healing due to diabetes.
[0004] Bodybuilders use citrulline malate, a formulation of
L-citrulline with malic acid added, as a supplement. It is
hypothesized that citrulline malate can increase NO and growth
hormone synthesis, and can therefore promote improvements in
athletic performance and physical energy levels. Such performance
enhancement effects of citrulline, however, lack clinical
validation.
SUMMARY
[0005] The present disclosure, in certain embodiments, provides
solid compositions and drinks that contain citrulline and taurine.
As the accompanying experimental examples show, citrulline was able
to enhance the physical performance of mice, and the addition of
taurine significantly and synergistically boosted the enhancement
even further. By contrast, gamma-aminobutyric acid (GABA), which
would have been thought to be more likely to synergize with
citrulline, did not even produce additive effects.
[0006] Also, even though taurine has a strong unpleasant astringent
bitter taste in solutions, when it was added to citrulline, it
unexpectedly improved the tastes of citrulline which on its own
only has a mild unnatural taste in solutions. When more taurine was
added (e.g., when there was more taurine than citrulline in the
solution) its astringent bitter taste overshadows the milder taste
of citrulline. Taurine's ability to improve the taste of citrulline
in solutions, at suitable weight ratios is another surprising
discovery of the present disclosure.
[0007] In one embodiment, therefore, the present disclosure
provides a composition comprising citrulline and taurine. In some
embodiments, the citrulline and the taurine have a weight ratio of
5:1 to 1:5, or 5:1 to 1:1, or 4.56:1 to 2.57:1. In some
embodiments, citrulline and taurine together constitute at least
20%, 30%, 40%, 50%, 55%, 60%, 70%, 80% or 90% w/w of the solid
composition or solid portion of the composition. In some
embodiments, the composition is a solid composition.
[0008] In some embodiment, the solid composition does not include
more than 30%, or 20% w/w of the combination of any other amino
acid, vitamins, peptides, proteins, herb extracts, fatty acids,
fibers, probiotics, glucosamine, chondroitin, and CoQ10. In some
embodiments, the solid composition does not include more than 10%
or 5% w/w malate. In some embodiments, the solid composition does
not include more than 10% or 5% w/w arginine. In some embodiments,
the solid composition does not include more than 30% of 20% w/w of
the combination of arginine, malate, niacin, agmatine, alanine,
gamma-aminobutyric acid (GABA), creatine, and dextrin.
[0009] In some embodiments, the solid composition does not include
more than 10% or 5% w/w of any other nutritional supplement or
activity-enhancing supplement. In some embodiments, the solid
composition does not include more than 1% w/w of any other
nutritional supplement or activity-enhancing supplement.
[0010] In some embodiments, the solid composition further comprises
caffeine. In some embodiments, the solid composition further
comprises a mineral. In some embodiments, the mineral is selected
from calcium, potassium, magnesium, iron, zinc, copper, chromium,
selenium, molybdenum, cobalt, nickel, vanadium, tin, strontium, or
rubidium.
[0011] In some embodiments, the citrulline and the taurine have a
weight ratio of 4:1 to 2.85:1. In some embodiments, the citrulline
and the taurine have a weight ratio of 3.55:1 to 3:1. In some
embodiments, the citrulline and the taurine have a weight ratio of
3.35:1 to 3.08:1. In some embodiments, the citrulline and the
taurine have a weight ratio of about 3.17:1.
[0012] In some embodiments, the citrulline and the taurine together
constitute at least 60% w/w of the solid composition. In some
embodiments, the citrulline and the taurine together constitute at
least 75% w/w of the solid composition.
[0013] In some embodiments, the solid composition further comprises
a sweetener, a stabilizer, a binder, a coloring agent, and/or an
anticaking agent.
[0014] In some embodiments, provided is a drink obtainable by
dissolving the solid composition of the present disclosure in water
or an aqueous solution.
[0015] Also provided, in some embodiments, are methods of using the
compositions for improving the athletic performance of a mammalian
subject. Also provided, in some embodiments, are methods of using
the compositions for or treating or preventing prehypertension,
hypertension, or erectile dysfunction (ED).
BRIEF DESCRIPTION OF THE DRAWINGS
[0016] The drawings described herein are for illustration purposes
only. The drawings are not intended to limit the scope of the
present disclosure.
[0017] FIG. 1 shows a lack of synergistic effect between citrulline
and gamma-aminobutyric acid (GABA) on exercise time to exhaustion.
Mice were given either normal saline (control), citrulline (200
mg/kg body weight), or GABA (150 mg/kg body weight), or
citrulline/taurine mixture (200 mg/kg+150 mg/kg) 1 hour prior to
the test. The mice swam with weights attached to their tails,
corresponding to 1/8 of their body weight, and the swimming time to
exhaustion were recorded. Values represent means.+-.SE.
[0018] FIG. 2 shows the synergistic effect of citrulline and
taurine on exercise time to exhaustion. Mice were given either
normal saline (control), citrulline (200 mg/kg body weight), or
taurine (200 mg/kg body weight), or citrulline/taurine mixture (200
mg/kg+200 mg/kg) 1 hour prior to the test. The mice swam with
weights attached to their tails, corresponding to 1/8 of their body
weight, and the swimming time to exhaustion were recorded. Values
represent means.+-.SE.
[0019] FIG. 3 shows the synergistic effect of citrulline and
taurine, on lower doses, on exercise time to exhaustion. Mice were
given either normal saline (control), or citrulline (75 mg/kg body
weight), or taurine (25 mg/kg body weight), or citrulline taurine
mixture (75 mg/kg+25 mg/kg) 1 hour prior to the test. The mice swam
with weights attached to their tails, corresponding to 1/8 of their
body weight, and the swimming time to exhaustion were recorded.
Values represent means.+-.SE.
[0020] FIG. 4 charts the results of a taste survey of various
drinks having different ratios of citrulline and taurine.
[0021] FIG. 5 charts the results of a second taste survey of
various drinks having different ratios of citrulline and
taurine.
[0022] Reference is now made in detail to certain embodiments of
the present disclosure. While certain embodiments of the present
disclosure are described, it will be understood that it is not
intended to limit the embodiments of the present disclosure to the
disclosed embodiments. To the contrary, reference to embodiments of
the present disclosure is intended to cover alternatives,
modifications, and equivalents as may be included within the spirit
and scope of the embodiments of the present disclosure as defined
by the appended claims.
DETAILED DESCRIPTION
[0023] The following description sets forth exemplary embodiments
of the present technology. It should be recognized, however, that
such description is not intended as a limitation on the scope of
the present disclosure but is instead provided as a description of
exemplary embodiments.
[0024] The potential ability of citrulline to boost endurance
performance was assessed with a loaded swimming test in mice. Mice
administered with 200 mg/kg citrulline exhibited an increase of
about 26% in endurance time on average (from about 194.7 seconds in
control to about 245.4 seconds). Another commonly used athletic
supplement, gamma-aminobutyric acid (GABA), was able to increase
the endurance time too, but at a more modest rate of 11.4%.
[0025] The addition of GABA to citrulline resulted in a slightly
higher effect than citrulline alone (31% vs. 26%). This slight
increase suggests no benefit of the combination given the small
benefit based on the almost doubled dosage (350 mg/kg vs. 200
mg/kg).
[0026] Surprisingly, when citrulline (200 mg/kg) and taurine (200
mg/kg) were used together, the combinatory effect almost tripled
each agent alone (63.1% in combination vs. 15.6% for citrulline and
23.6% for taurine alone). This is clear indication of synergism,
which is surprising and unexpected on its own, and more so because
of the lack of synergism between citrulline and GABA. Moreover, the
magnitude of the synergism (63.1% observed vs. a 35.5% theoretical
synergism threshold) is even more surprising.
[0027] At a greatly reduced dose of 25 mg/kg, the effect of taurine
alone was less pronounced (181 seconds as compared to 168.3 seconds
in controls, a mere 7.5% increase). The effect of citrulline, at 75
mg/kg, was also less significant (207.7 seconds). Here, the
increases by citrulline and taurine were 23.4% and 7.5%,
respectively. The combination treatment, however, resulted in a
53.7% increase, which further underscores the synergism between
these two amino acids.
[0028] Taurine, however, has an unpleasant astringent bitter taste
in a solution. The addition of taurine to citrulline, therefore,
may not be a welcome change, even though citrulline itself has a
sense of unnatural sweetness. In another surprising discovery of
the present disclosure, an aqueous solution of citrulline and
taurine in a weight ratio of about 2.5:1 to about 4.5:1, in
particular at about 3.17:1, has the best taste. More strikingly,
the taste of the mixture at the suitable ratio is better than each
of citrulline and taurine alone. In other words, citrulline and
taurine can neutralize each other's unpleasant flavor, and this was
entirely unexpected.
[0029] In accordance within one embodiment of the present
disclosure, therefore, provided is a solid composition comprising
citrulline and taurine. In some embodiments, a solution that
contains the solid composition dissolved in the solution is also
provided. Methods of preparing the solid composition or the
solution are also provided, in various embodiments.
[0030] In some embodiments, the citrulline and the taurine have a
weight ratio from about 5:1 to about 1:5. In some embodiments, the
weight ratio between citrulline and the taurine is from about 5:1,
4.5:1, 4:1, 3.5:1, 3:1, 2.5:1, 2:1 1.5:1, 1:1, 0.9:1, 0.8:1, 0.7:1,
0.6:1, 0.5:1, 0.4:1, 0.3:1, 0.25:1, or 0.2:1 to about 1:5, 1:4.5,
1:4, 1:3.5, 1:3, 1:2.5, 1:2, 1:1.5, 1:1, 1:0.9, 1:0.8, 1:07, 1:0.6,
1:0.5, 1:0.4, 1:0.3 or 1:02.
[0031] In some embodiments, the citrulline and the taurine have a
weight ratio of about 4.56:1 to about 2.57:1. In some embodiments,
the citrulline and the taurine have a weight ratio of about 4:1 to
about 2.85:1. In some embodiments, the citrulline and the taurine
have a weight ratio of about 3.55:1 to about 3:1. In some
embodiments, the citrulline and the taurine have a weight ratio of
about 3.35:1 to about 3.08. In some embodiments, the citrulline and
the taurine have a weight ratio of about 3.17.
[0032] In some embodiments, the citrulline and the taurine have the
weight percentages or weight ratios as shown in Table 1.
TABLE-US-00001 TABLE 1 Preferred Taurine % or Citrulline/Taurine
Ratios Weight % of taurine Weight Ratio in total
(citrulline/taurine) low high low high Preferred 18 28 4.56 2.57
More Preferred 20 26 4.00 2.85 Even More Preferred 22 25 3.55 3.00
Most Preferred 23 24.5 3.35 3.08
[0033] In some embodiments, the citrulline and the taurine in
combination constitute the substantial portion of the solid
composition. In some embodiments, the citrulline and the taurine in
combination constitute at least about 20%, 30%, 40%, 50%, 60%, 70%,
75%, 80%, 85%, 90%, 95%, 98%, or 99% of the solid composition.
[0034] In some embodiments, the composition does not include a
significant portion of other commonly used nutritional or
parametrical active agents. Examples of nutritional or parametrical
active agents include other amino acids, vitamins, peptides,
proteins, herb extracts, fatty acids, fibers, probiotics,
glucosamine, chondroitin, and CoQ10. In some embodiments, the
composition contains less than about 40%, 35%, 30%, 25%, 20%, 15%,
10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2% or 1% (w/w) of other
nutritional or parametrical active agents. In some embodiments, the
composition contains less than about 40%, 35%, 30%, 25%, 20%, 15%,
10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2% or 1% (w/w) of other amino
acids.
[0035] In some embodiments, the composition contains less than
about 40%, 35%, 30%, 25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%,
3%, 2% or 1% (w/w) malate. In some embodiments, the composition
contains less than about 40%, 35%, 30%, 25%, 20%, 15%, 10%, 9%, 8%,
7%, 6%, 5%, 4%, 3%, 2% or 1% (w/w) arginine. In some embodiments,
the composition contains less than about 40%, 35%, 30%, 25%, 20%,
15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2% or 1% (w/w) the
combination of arginine, malate, niacin, agmatine, alanine and
dextrin. In some embodiments, the composition contains less than
about 40%, 35%, 30%, 25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%,
3%, 2% or 1% (w/w) creatine.
[0036] In some embodiments, the composition contains less than
about 40%, 35%, 30%, 25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%,
3%, 2% or 1% (w/w) other activity/performance-enhancing
supplements.
[0037] The composition can further include one or more other
excipients or ingredients. Non-limiting examples of excipients
include diluents, disintegrants, binders, lubricants, sweeteners,
flavoring agents and anti-cake agents.
[0038] Diluents act as fillers in nutraceutical/pharmaceutical
tablets to increase weight and improve content uniformity. Examples
include, without limitation, microcrystalline cellulose, powdered
cellulose, pregelatinized starch, starch, lactitol, mannitol,
sorbitol, maltodextrin and combinations thereof.
[0039] Disintegrants are agents added to solid formulations (e.g.,
tablets) to promote the breakup of the tablet (or capsule "slugs")
into smaller fragments in an aqueous environment thereby increasing
the available surface area and promoting a more rapid release of
the drug substance. Examples include, without limitation,
croscarmellose sodium, crospovidone, starch, sodium carboxymethyl
starch, low-substituted hydroxypropyl cellulose and combinations
thereof.
[0040] Binders are formulated to act as an adhesive to bind
together powders, granules and other dry ingredients to impart to
the product the necessary mechanical strength. Examples include,
without limitation, distilled water, ethanol, hypromellose, sodium
carboxymethylcellulose, povidone, ethyl cellulose and combinations
thereof.
[0041] Lubricants are agents added to tablet and capsule
formulations to improve the powder processing properties of the
formulation. Lubricants (glidants) can enhance the powder flow by
reducing the inter-particle friction. Non-limiting examples include
sodium stearyl fumarate, magnesium stearate, calcium stearate,
stearic acid, glyceryl behenate, and combinations thereof.
[0042] Non-limiting examples of sweeteners include xylitol,
sorbitol, mannitol, maltitol, lactitol, isomalt, erythritol,
aspartame, acesulfame potassium, alitame, sodium saccharin, three
Sucralose, neotame, cyclamate, and combinations thereof.
[0043] Non-limiting examples of flavoring agents include orange
flavor, orange flavor, mango flavor, peach flavor, grapefruit
flavor, and combinations thereof.
[0044] Anti-caking agents are additives placed in powdered or
granulated materials, such as table salt, to prevent the formation
of lumps and for easing packaging, transport, and consumption.
Anticaking agents function either by adsorbing excess moisture, or
by coating particles and making them water repellent. Non-limiting
examples of anti-caking agents include silica, tricalcium
phosphate, microcrystalline cellulose and combination thereof.
[0045] In addition, in effervescent tablet or granules, an agent
that produces carbon dioxide Is needed, and their examples include
acidic materials such as citric acid, tartaric acid, fumaric acid,
malic acid, adipic acid, succinic acid, ascorbic acid, maleic acid,
and combinations thereof. Further included in the effervescent
agent are alkali metal hydrogencarbonates including sodium
hydrogencarbonate, potassium hydrogencarbonate, and combinations
thereof.
[0046] In some embodiments, the composition further includes
caffeine. In some embodiments, the composition further includes a
mineral, such as calcium, potassium, magnesium, iron, zinc, copper,
chromium, selenium, molybdenum, cobalt, nickel, vanadium, tin,
strontium, or rubidium.
[0047] The solid compositions of the present disclosure can be
provided as tablets, capsules, powders, granules, or effervescent
tablets or granules. Solutions (e.g., beverages) that include
citrulline and the taurine are also provided.
[0048] The solutions can be prepared by dissolving a solid
composition of the present disclosure in water or an aqueous
solution. In some embodiments, the solution includes from about
0.1% (w/w) to about 20% (w/w) of a combination of citrulline and
the taurine. In some embodiments, the solution includes at least
about 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%,
1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, or 2% (w/w)
of the combination of citrulline and the taurine. In some
embodiments, the solution includes no more than about 20%, 18%,
15%, 12%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2.9%, 2.8%, 2.7%, 2.6%,
2.5%, 2.4%, 2.3%, 2.2%, 2.1%, 2%, 1.9%, 1.8%, 1.7%, 1.6%, 1.5%,
1.4%, 1.3%, 1.2%, 1.1%, or 1% of the combination of citrulline and
the taurine.
[0049] In some embodiments, the solution has a pH of about 2.0 to
about 9.0. In some embodiments, the solution has a pH of at least
about 2.0, 2.5, 3.0, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, or 7. In some
embodiments, the solution has a pH of not higher than about 9, 8.5,
8, 7.5, 7, 6.5, 6, 5.5, 5, 4.5 or 4. In some embodiments, the pH is
from about 2.5 to about 3.5. In some embodiments, the pH is from
about 3 to 4. In some embodiments, the pH is from about 3.5 to 4.5.
In some embodiments, the pH is from about 4 to 5. In some
embodiments, the pH is from about 5 to 6. In some embodiments, the
pH is from about 6 to 7. In some embodiments, the pH is from about
7 to 8. In some embodiments, the pH is from about 8 to 9.
[0050] In some embodiments, the citrulline and the taurine in the
solution have a weight ratio of about 4.56:1 to about 2.57:1. In
some embodiments, the citrulline and the taurine in the solution
have a weight ratio of about 4:1 to about 2.85:1. In some
embodiments, the citrulline and the taurine in the solution have a
weight ratio of about 3.55:1 to about 3:1. In some embodiments, the
citrulline and the taurine in the solution have a weight ratio of
about 3.35:1 to about 3.08. In some embodiments, the citrulline and
the taurine in the solution have a weight ratio of about 3.17.
[0051] Specific examples of solid compositions and solutions
disclosed here include, without limitation, those provided in
Experimental Example 5.
[0052] Methods for preparing and using the aqueous solutions of the
present disclosure are also provided. In some embodiments, the
solutions can be prepared by adding each of the ingredients into a
water-based solution.
[0053] In various embodiments, the solid compositions and solutions
disclosed herein can be used in methods for enhancing stamina,
athletic performance, endurance, and/or reducing muscle soreness,
fatigue or cramping in a subject in need thereof. In some
embodiments, the solid compositions and solutions disclosed herein
can be used in methods for preventing or treating hypertension or
prehypertension in a subject in need thereof. In some embodiments,
the solid compositions and solutions disclosed herein can be used
in methods for preventing or treating erectile dysfunction (ED) in
a male subject in need thereof. The method, in some embodiments,
entails orally administering to the subject an effective amount of
the solid composition or solution of the present disclosure.
[0054] In some embodiments, provided is a method or use for
improving the athletic performance of a mammalian subject. In some
embodiments, provided is a method or use for enhancing the physical
endurance or reducing fatigue of a mammalian subject. In some
embodiments, provided is a method or use for enhancing the muscular
strength of a mammalian subject.
[0055] In some embodiments, provided is a method or use for
treating or preventing erectile dysfunction (ED) in a male
mammalian subject. In some embodiments, wherein the ED is mild or
moderate. In some embodiments, provided is a method or use for
treating or preventing hypertension or prehypertension in a
mammalian subject.
[0056] In various embodiments, the method entails administering, in
particular orally, to the subject a solid composition or a drink of
the disclosure. In some embodiments, about 0.1 g to about 100 g of
the combination of citrulline and the taurine are administered
daily. In some embodiments, the daily dose of the combination of
citrulline and the taurine is at least 0.1 g, 0.2 g, 0.3 g, 0.4 g,
0.5 g, 0.6 g, 0.7 g, 0.8 g, 0.9 g, 1 g, 2 g, 3 g, 4 g, 5 g, 6 g, 7
g, 8 g, 9 g, 10 g, 15 g, 20 g, 30 g, 40 g, or 50 g. In some
embodiments, the daily dose of the combination of citrulline and
the taurine is not greater than 100 g, 90 g, 80 g, 70 g, 60 g, 50
g, 40 g, 30 g, 20 g, 15 g, 10 g, 9 g, 8 g, 7 g, 6 g, 5 g, 4 g, 3 g,
2 g or 1 g.
[0057] In some embodiments, the solid composition or the drink is
administered once, twice or three times daily. In some embodiments,
the solid composition or the drink is administered prior to,
during, or following physical activities, or prior to a sexual
activity.
[0058] In some embodiments, the administration follows an intense
physical activity by the subject. In one embodiment, the
administration is made before an intense physical activity by the
subject.
[0059] In some embodiments, the effective amount of the solution is
about 30 ml, 50 ml, 75 ml, 100 ml, 120 ml, 150 ml, 200 ml, 250 ml,
300 ml, 400 ml, or 500 ml, without limitation. In some embodiments,
the effective amount of the solid composition is about 0.1, 0.2,
0.5 g, 1 g, 1.5 g, 2 g, 3 g, 4 g, 5 g, 10 g, 15 g or 20 g without
limitation.
[0060] In some embodiments, the subject experiences muscle
soreness, fatigue, or cramping. In some embodiments, the subject,
following the administration, experiences reduced muscle soreness,
fatigue or cramping. In some embodiment, the subject desires the
flavor of the solution.
Examples
[0061] The following examples are included to demonstrate specific
embodiments of the disclosure. It should be appreciated by those of
skill in the art that the techniques disclosed in the examples
which follow represent techniques to function well in the practice
of the disclosure, and thus can be considered to constitute
specific modes for its practice. However, those of skill in the art
should, in light of the present disclosure, appreciate that many
changes can be made in the specific embodiments which are disclosed
and still obtain a like or similar result without departing from
the spirit and scope of the disclosure.
Example 1. Effects of Citrulline and GABA on Endurance and Muscle
Strength
[0062] This example tested the effect of citrulline on the
endurance and muscle strength of mice in exercise, and explores the
potential synergistic effect of gamma-aminobutyric acid (GABA) to
citrulline.
Materials and Methods
[0063] Animals. Adult (8-10 weeks) male ICR mice were used in this
study and purchased from Beijing Vital River Laboratory Animal
Technology Co., Ltd. All the mice were provided with a normal diet
and water ad libitum. All procedures were approved by institutional
guidelines and the Animal Care and Use Committee (Huazhong
University of Science and Technology, Wuhan, China) of the
university's animal core facility. The mice were group-housed under
a 12 h light-dark cycle (lights on at 8:00 a.m.), at consistent
humidity (50.+-.5%), and ambient temperature (22.about.24.degree.
C.).
[0064] The animals were bred with three mice per cage in the
laboratory, and were acclimatized for 1 week before the
experiments. The mice were grouped randomly for each experiment.
The mice in each group were given the corresponding dose amino acid
and the control group was treated with the equal volume of normal
saline (NS) through intragastric administration. Loaded swimming
experiments were performed to assess the acute effect of the amino
acids on endurance one hour after the treatment
[0065] The NS injection were purchased from Beijing Pharmaceutical
Group Co., Ltd. The amino acid powders were dissolved in NS and
then were intragastric administration to mice by lavage needle
(Beijing Vital River Laboratory Animal Technology Co., Ltd.).
Loaded Swimming Test
[0066] Acute effects on endurance performance were assessed by
loaded swimming test one hour after the amino acid treatment. The
swimming exercise was carried out in a tank (diameter 120 cm)
filled with fresh water to 30-cm depth at 24.+-.2.degree. C. A tin
wire (1/8 of body weight) was attached on the tail of the mouse.
Each mouse was considered to have reached exhaustion when it failed
to rise its face to the surface of the water for inhale within 5 s
period. The length of time each mouse kept swimming before
exhaustion (exercise time to exhaustion) was recorded. At the end
of the session, the mice were removed from the water, dried with
paper towels, and placed back in their home cages.
Results
[0067] Sixty mice were randomly divided into four groups (n=15 per
group). Each of the four group were treated with NS, citrulline
(200 mg/kg body weight), GABA (150 mg/kg body weight), and
citrulline/GABA mixture (200 mg/kg+150 mg/kg) respectively. The
results were summarized below (Table 2 and FIG. 1).
TABLE-US-00002 TABLE 2 Effects of citrulline and citrulline/GABA
combination on exercise time to exhaustion Control Citrulline GABA
Citrulline + GABA Average Time (s) 194.7 245.4 216.8 255 SE 14.67
28.01 37.88 23.03 Increase -- 26% 11.4% 31%
[0068] Citrulline increased the endurance time, as compared to
vehicle control, by about 26%. GABA alone also increased the
endurance, though at a slightly lower rate, at about 11.4%. The
combination of citrulline and GABA resulted in a 31% increase.
[0069] A commonly accepted way to estimate the combinatory effect,
without synergism, is Ec=Ea+Eb-Ea*Eb, where Ea, Eb and Ec represent
the effects of agents a and b and their combination, respectively.
Here, the estimated combinatory effect without synergism would be a
34.4% increase (vs. a sum of both values being 37.4%). In FIG. 1,
the predicted synergism threshold (261.7 seconds) is indicated as a
horizontal dotted line. The observed combinatory effect was an
endurance enhancement of 31% (255 seconds), suggesting that there
was no synergism between citrulline and GABA. Also, given the
modest added effect of GABA (5% over citrulline's 26%) from the 150
mg/kg dose, GABA does not appear to be promising additive to
citrulline.
[0070] This result was disappointing as GABA was suggested to have
the ability to enhance muscle building. Further, according to
Shyamaladevi et al., Brain Research Bulletin 57(2):231-6 (2002),
for instance, nitric oxide (NO) increases the blood-brain barrier
permeability for GABA. Meanwhile, citrulline boosts NO production
in the body. Therefore, when citrulline is supplied to the body, it
would have been expected to raise the availability of GABA in the
brain. The interaction between the two agents, therefore, would
have been expected to exhibit synergy. Such synergy, however, was
not observed.
Example 2. Unexpected Synergy Between Citrulline and Taurine on
Endurance and Muscle Strength
[0071] This example continued to look for agents that could
significantly further increase citrulline's activity in enhancing
the endurance and muscle strength of mice in exercise. Taurine was
surprisingly found to have such synergism with citrulline.
[0072] The methods used here are the same as in Example 1, but GABA
(150 mg/kg) was replaced by taurine (200 mg/kg). The results were
summarized below (Table 3 and FIG. 2).
TABLE-US-00003 TABLE 3 Effect of citrulline taurine combination on
exercise time to exhaustion Control Citrulline Taurine Citrulline +
Taurine Average Time (s) 168.3 194.6 208 274.5 SE 12.58 17.02 22.20
20.33 Increase -- 15.6% 23.6% 63.1%
[0073] Citrulline and taurine increased the endurance time by 15.6%
and 23.6%, respectively. As the synergism estimation methodology
explained in Example 1, the estimated combinatory effect without
synergism would be a 35.5% increase (vs. a sum of both values being
39.2%). In FIG. 2, the predicted synergism threshold (228.1
seconds) is indicated as a horizontal dotted line. The observed
combinatory effect, however, was an endurance enhancement of 63.1%
(274.5 seconds), clearly indicating a strong synergy between these
two amino acids.
Example 3. Synergy Between Lower Doses of Citrulline and Taurine on
Endurance and Muscle Strength
[0074] This example tested whether the synergism between citrulline
and taurine is still present in lower doses, and confirmed the
synergism.
[0075] The methods used here are the same as in Example 1. Sixty
mice were randomly divided into four groups (n=15 per group). Each
of the four group were treated with NS, citrulline (75 mg/kg body
weight), taurine (25 mg/kg body weight), and citrulline taurine
mixture (75 mg/kg+25 mg/kg) respectively. The results were
summarized below (Table 4 and FIG. 3).
TABLE-US-00004 TABLE 4 Effect of lower doses of citrulline taurine
combination on exercise time to exhaustion. Control Citrulline
Taurine Citrulline + Taurine Average Time (s) 168.3 207.7 181 258.7
SE 18.53 16.67 16.56 22.05 Increase -- 23.4% 7.5% 53.7%
[0076] Here, the lower doses of citrulline and taurine increased
the endurance time by 23.4% and 7.5%, respectively. The estimated
combinatory effect without synergism would be a 29.1% increase (vs.
a sum of both values being 30.9%). In FIG. 3, the predicted
synergism threshold (217.4 seconds) is indicated as a horizontal
dotted line. The observed combinatory effect, however, was an
endurance enhancement of 53.7% (258.7 seconds), again a showing of
strong synergy between these two amino acids even at the lowered
doses.
[0077] Further, Examples 2 and 3 demonstrate that the synergism
between citrulline and taurine exist at different dose levels and
at different ratios (e.g., 1:1 and 3:1).
Example 4. Taste Surveys
[0078] Citrulline solution does not have a strong taste, but has a
sense of unnatural sweetness that is distinguishable. The taurine
solution, however, clearly has an astringent bitter taste, even at
a fairly diluted concentration. The mixture of citrulline and
taurine, therefore, presents a challenge in terms of flavoring, as
compared to citrulline alone. This example explored options to
minimize the unpleasant taste with different ratios of these two
amino acids.
Taste Survey No. 1
Method
[0079] Six different citrulline-taurine mixtures were prepared with
citrulline/taurine ratios ranging from 8/0, 7/1, 4/1, 6/2, 5/3, to
4/4. Testing samples were prepared by dissolving 6 grams of each
mixture in 250 ml drinking water separately. Thirty volunteers, 12
females and 18 males, aged from 22-55 participated in the test. The
volunteers were asked to taste each sample without knowing the
composition of the samples. Each volunteer sorted the samples from
1 to 6, with 1 being the most liked and 5 being the least liked
sample.
Results
[0080] The survey results are shown in Table 5.
TABLE-US-00005 TABLE 5 Survey results Citrulline:Taurine Ratio
Individual 8:0 7:1 4:1 6:2 5:3 4:4 P01 6 5 4 3 1 2 P02 4 3 2 1 6 5
P03 4 3 1 2 5 6 P04 6 3 2 1 4 5 P05 4 3 1 2 5 6 P06 5 2 3 1 4 6 P07
6 4 3 1 2 5 P08 5 6 4 2 1 3 P09 4 3 2 1 5 6 P10 5 3 2 1 4 6 P11 3 4
5 1 2 6 P12 6 4 2 1 3 5 P13 4 3 2 1 6 5 P14 2 3 4 1 5 6 P15 4 2 1 3
6 5 P16 6 5 3 1 2 4 P17 3 4 2 1 5 6 P18 6 4 5 3 1 2 P19 4 2 6 5 3 1
P20 1 3 5 6 2 4 P21 3 4 2 5 1 6 P22 6 3 1 2 4 5 P23 4 3 1 2 6 5 P24
2 4 3 1 5 6 P25 4 1 3 2 6 5 P26 5 4 1 2 3 6 P27 4 3 1 2 5 6 P28 4 1
3 5 2 6 P29 4 3 1 2 5 6 Total 124 95 75 61 109 145
[0081] The results are plotted in FIG. 4, which indicates that when
citrulline and taurine were mixed at about a 3:1 weight ratio, the
taste was the most favorable. What was entirely unexpected and even
striking is that the mixtures had more favorable tastes than each
of its ingredients alone. For instance, citrulline alone (i.e., 8:0
citrulline to taurine) had a taste survey score of 124. When
taurine was added, the taste survey score improved to 95 (7:1), 75
(4:1) and 61 (6:2) and then deteriorated when more taurine was
added. In other words, addition of the more unfavorable taurine was
able to even reduce the less unfavorable taste of citrulline.
[0082] To further investigate the tastes of citrulline taurine
combinations, nine different citrulline-taurine mixtures were
prepared with taurine content in the mixture of 20%, 22%, 24%, 25%,
26%, 28%, 30%, 32%, and 34%. Testing samples were prepared by
dissolving 6 grams of each mixture in 250 ml drinking water
separately. Twenty volunteers, 11 females and 9 males, aged from
22-55 participated in the test. The volunteers were asked to taste
each sample without knowing the composition of the samples. Each
volunteer sorted the samples from 1 to 9, with 1 being the most
liked and 5 being the least liked sample. The survey results are
provided in Table 6 below.
TABLE-US-00006 TABLE 6 Survey results Indi- Taurine content in the
mixture vidual 20% 22% 24% 25% 26% 28% 30% 32% 34% Q01 8 2 1 6 9 7
5 4 3 Q02 1 2 3 5 9 4 6 8 7 Q03 3 2 1 4 6 7 8 9 5 Q04 9 8 1 7 2 6 4
3 5 Q05 2 3 1 4 5 7 6 9 8 Q06 4 3 5 1 2 6 7 9 8 Q07 4 1 2 3 5 7 6 9
8 Q08 2 1 4 5 3 6 7 8 9 Q09 5 4 2 3 1 6 7 8 9 Q10 5 2 1 3 4 7 6 8 9
Q11 3 1 2 4 5 6 7 9 8 Q12 7 6 5 2 9 4 3 1 8 Q13 3 1 2 4 5 6 7 9 8
Q14 7 6 5 4 3 2 1 8 9 Q15 3 1 2 6 5 4 8 7 9 Q16 4 3 2 1 5 6 7 9 8
Q17 5 2 3 1 4 7 6 8 9 Q18 5 4 2 3 1 6 7 8 9 Q19 3 2 4 1 8 6 9 7 5
Total 83 54 48 67 91 110 117 141 144
[0083] As the composition difference between samples gets smaller,
the curve became shallower. However, the result indicates that
samples with taurine content between 20% and 28% were best "liked",
with 24% being the winner (FIG. 5).
Example 5. Production Samples
[0084] This example lists a number of production samples in
different forms (solid or in solution).
Production Sample 1 (Powder):
TABLE-US-00007 [0085] Ingredient Amount Unit Citrulline 76 parts
Taurine 24 parts Maltodextrin 1.5 parts Microcrystalline cellulose
1 parts Food flavor 1 parts
[0086] Production sample 1 is prepared as follows. The ingredients
are weighed according to the formula, and are mixed together in a
uniform manner. Alternatively, they are added into water is then
spray dried.
Production Sample 2 (Oral Solution):
TABLE-US-00008 [0087] Ingredient Amount Unit Citrulline 1.52 %
Taurine 0.48 % Creatine monohydrate 0.15 % Maltodextrin 0.1 %
Concentrated lemon juice 0.005 % Sodium cyclamate 0.0034 %
Acesulfame 0.00255 % Sodium bicarbonate 0.001 % Aspartame 0.00085 %
Orange flavor 0.0005 % Water q.s. to 100 mg %
[0088] Production sample 2 is prepared as follows. The ingredients
are weighed according to the formula, stirred and dissolved
uniformly to obtain a mixed liquid. The mixture is filtered and
then bottled. Sterilization is carried out to obtain a finished
product.
Production Sample 3 (Tablets or Granules):
TABLE-US-00009 [0089] Ingredient Amount Unit Citrulline 80 parts
Taurine 20 parts Filler 25 parts Disintegrant 2.5 parts Lubricant
2.5 parts Binder 2.5 parts Sweetener 0.1 parts
[0090] Production sample 3 is prepared as follows. The ingredients
are weighed according to the formula and are mixed with water in a
wet granulator. The particles are then dried in an oven to obtain
granules. The granules can be pressed to prepare tablets.
Production Sample 4 (Effervescent Granules):
TABLE-US-00010 [0091] Ingredient Amount Unit Citrulline 66 parts
Taurine 33 parts Filler 25 parts Citric acid 25 parts Sodium
bicarbonate 25 parts
[0092] Production sample 4 is prepared as follows. The ingredients
are weighed according to the formula and are mixed with water in a
wet granulator. The particles are then dried in an oven to obtain
effervescent granules.
Production Sample 5 (Effervescent Tablets):
TABLE-US-00011 [0093] Ingredient Amount Unit Citrulline 75 parts
Taurine 25 parts Filler 25 parts Disintegrant 5 parts Lubricant 5
parts Sweetener 2.5 parts Citric acid 25 parts Sodium bicarbonate
25 parts
[0094] Production sample 5 is prepared as follows. The ingredients
(except citric acid and sodium bicarbonate) are weighed according
to the formula and are mixed with water in a wet granulator. The
particles are then dried in an oven. Citric acid and sodium
bicarbonate are then added to the particles and pressed to prepare
effervescent tablets.
Production Sample 6 (Capsules):
TABLE-US-00012 [0095] Ingredient Amount Unit Citrulline 76 parts
Taurine 24 parts Filler 25 parts Disintegrant 5 parts Lubricant 5
parts Binder 5 parts
[0096] Production sample 6 is prepared as follows. The ingredients
(except citric acid and sodium bicarbonate) are weighed according
to the formula and are mixed with water in a wet granulator. The
particles are then dried in an oven and filled into capsules in a
filling machine.
[0097] Unless otherwise defined, all technical and scientific terms
used herein have the same meaning as commonly understood by one of
ordinary skill in the art to which this invention belongs.
[0098] The inventions illustratively described herein may suitably
be practiced in the absence of any element or elements, limitation
or limitations, not specifically disclosed herein. Thus, for
example, the terms "comprising", "including," "containing", etc.
shall be read expansively and without limitation. Additionally, the
terms and expressions employed herein have been used as terms of
description and not of limitation, and there is no intention in the
use of such terms and expressions of excluding any equivalents of
the features shown and described or portions thereof, but it is
recognized that various modifications are possible within the scope
of the invention claimed.
[0099] Thus, it should be understood that although the present
invention has been specifically disclosed by preferred embodiments
and optional features, modification, improvement and variation of
the inventions embodied therein herein disclosed may be resorted to
by those skilled in the art, and that such modifications,
improvements and variations are considered to be within the scope
of this invention. The materials, methods, and examples provided
here are representative of preferred embodiments, are exemplary,
and are not intended as limitations on the scope of the
invention.
[0100] The invention has been described broadly and generically
herein. Each of the narrower species and subgeneric groupings
falling within the generic disclosure also form part of the
invention. This includes the generic description of the invention
with a proviso or negative limitation removing any subject matter
from the genus, regardless of whether or not the excised material
is specifically recited herein.
[0101] In addition, where features or aspects of the invention are
described in terms of Markush groups, those skilled in the art will
recognize that the invention is also thereby described in terms of
any individual member or subgroup of members of the Markush
group.
[0102] All publications, patent applications, patents, and other
references mentioned herein are expressly incorporated by reference
in their entirety, to the same extent as if each were incorporated
by reference individually. In case of conflict, the present
specification, including definitions, will control.
[0103] It is to be understood that while the disclosure has been
described in conjunction with the above embodiments, that the
foregoing description and examples are intended to illustrate and
not limit the scope of the disclosure. Other aspects, advantages
and modifications within the scope of the disclosure will be
apparent to those skilled in the art to which the disclosure
pertains.
* * * * *