U.S. patent application number 14/541278 was filed with the patent office on 2015-03-12 for preservation of ergothioneine.
This patent application is currently assigned to ELC Management LLC. The applicant listed for this patent is ELC Management LLC. Invention is credited to Daniel B. Yarosh.
Application Number | 20150073027 14/541278 |
Document ID | / |
Family ID | 42396282 |
Filed Date | 2015-03-12 |
United States Patent
Application |
20150073027 |
Kind Code |
A1 |
Yarosh; Daniel B. |
March 12, 2015 |
Preservation of Ergothioneine
Abstract
Compositions comprising ergothioneine and a trimethylamine
absorber are provided. Also provided are methods for preventing,
reducing or minimizing the fishy, amine odor, due to
trimethylamine, that is associated with the processing and/or
storage of a preparation containing ergothioneine, by combining
with the ergothioneine, during processing or prior to storage, a
trimethylamine absorber in an amount sufficient to prevent the
detection of any trimethylamine odor by the human nose. A method is
further provided for ameliorating the methylamine odor associated
with an aqueous ergothioneine-containing preparation after it has
developed a fishy trimethylamine odor.
Inventors: |
Yarosh; Daniel B.; (Merrick,
NY) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
ELC Management LLC |
Melville |
NY |
US |
|
|
Assignee: |
ELC Management LLC
Melville
NY
|
Family ID: |
42396282 |
Appl. No.: |
14/541278 |
Filed: |
November 14, 2014 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
12362594 |
Jan 30, 2009 |
8410156 |
|
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14541278 |
|
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Current U.S.
Class: |
514/392 |
Current CPC
Class: |
A61K 8/365 20130101;
A61K 8/36 20130101; A61P 17/00 20180101; A61K 8/23 20130101; A61K
8/19 20130101; A61K 31/4172 20130101; A61K 8/676 20130101; A61K
47/12 20130101; A61K 8/4946 20130101; A61K 2300/00 20130101; A61K
31/4172 20130101; A61K 31/417 20130101; A61K 8/24 20130101; A61Q
19/00 20130101; A61K 45/06 20130101; A61K 9/0014 20130101 |
Class at
Publication: |
514/392 |
International
Class: |
A61K 47/12 20060101
A61K047/12; A61K 9/00 20060101 A61K009/00; A61K 31/417 20060101
A61K031/417 |
Claims
1. An aqueous-containing topical, cosmetic or pharmaceutical
composition having a neutral to acidic pH and comprising greater
than 20 .mu.M (w/v) ergothioneine and at least one trimethylamine
absorber comprising a weak acid, wherein the ergothioneine and the
trimethylamine absorber are present in the composition in amounts
sufficient such that trimethylamine gas released from the
ergothioneine during storage of the composition at room temperature
is converted by the trimethylamine absorber to a form which cannot
volatilize.
2. The aqueous-containing composition of claim 1, wherein the weak
acid has an acid dissociation constant (K.sub.a) of between about
1.8.times.10.sup.-16 and 55.5.
3. The aqueous-containing composition of claim 2, wherein the weak
acid is an organic acid selected from the group consisting of
acetic acid, ascorbic acid, citric acid, formic acid, heptanoic
acid, hexanoic acid, lactic acid, octanoic acid, oxalic acid,
pentanoic acid, propanoic acid, uric acid, and mixtures
thereof.
4. The aqueous-containing composition of claim 3, wherein the weak
organic acid is citric acid.
5. The aqueous-containing composition of claim 2, wherein the weak
acid is an inorganic acid selected from the group consisting of
boric acid, carbonic acid, chromic acid, hydrocyanic acid,
hydrofluoric acid, nitrous acid, phosphoric acid, sulfuric acid,
sulfurous acid and mixtures thereof.
6. The aqueous-containing composition of claim 1, wherein the weak
acid and the ergothioneine are present in the composition in a
ratio of greater than 1:1.
7. The aqueous-containing composition of claim 6, wherein the weak
acid and the ergothioneine are present in the composition in a
ratio of in the range of from about 1:1 to about 6:1.
8. The aqueous-containing composition of claim 7, wherein the weak
acid and the ergothioneine are present in the composition in a
ratio of in the range of from about 2:1 to about 4:1.
9. The aqueous-containing composition of claim 1, wherein
ergothioneine is present at a concentration of greater than 20 mM
(w/v).
10. The aqueous-containing composition of claim 1, which is in the
form of a solution, a suspension or an emulsion.
Description
[0001] The present application is a divisional of U.S. Ser. No.
12/362,594, filed Jan. 30, 2009.
FIELD OF THE INVENTION
[0002] The present invention relates to the preparation of
ergothioneine or ergothioneine-containing mixtures that ameliorates
changes in the odor of the product during processing and/or
storage. In particular, the preparation involves the addition of
acids and/or sulfur dioxide donors that reduce or prevent the
formation of trace volatile compounds to which the human nose is
particularly sensitive. The resulting preparations retain an
innocuous odor even after challenge with conditions, such as alkali
or storage over time, which would otherwise produce fishy, amine
odors in ergothioneine solutions or ergothioneine-containing
mixtures. The acids and/or sulfur dioxide donor compounds may also
be added to the ergothioneine solutions or ergothioneine-containing
mixtures after the detection of the odor to reduce or eliminate the
volatile trace compounds to which the human nose is sensitive.
BACKGROUND OF THE INVENTION
[0003] Ergothioneine is a non-essential L-amino acid found
naturally in the body. It is a very strong antioxidant, and because
it is a carnitine analog, it may also have activity in aerobic ATP
production. It is synthesized by types of fungi but not mammals,
who must acquire it in their diet. They do so by direct consumption
of fungi such as mushrooms, or from grains which have themselves
taken up ergothioneine from fungi in their roots. Methods for the
laboratory preparation of synthetic ergothioneine have been
described in, for example, H. Heath, A. Lawson and C. Rimington.
"2-Mercaptoglyoxalanes. Part I. The synthesis of Ergothioneine", J.
Chem Soc. (1951) pp 2215-2217 and in U.S. Pat. No. 5,438,151.
[0004] It has been observed that ergothioneine is quite stable at
acidic pH. At pH 5 a 0.05% solution remains within 1% of its
concentration after heating 40 C for 60 days. However, the reports
of its stability in alkali are mixed in the literature. See, for
example, Philip Hartman. "Ergothioneine as antioxidant", Methods
Enzymol. 186:310-318, 1990 at p. 311 (" . . . the sulfur atom of
Ergothioneine is remarkably stable to alkali"); H. Heath and G.
Toennies. "The preparation and properties of Ergothioneine
disulphide", J. Chem. Soc. pages 204-210, (1958) at p. 204 ("the
sulphur is completely unaffected by boiling 50% aqueous potassium
hydroxide."); Oxis International, Inc. Compound Monograph
"L-Ergothioneine. Revision III", at p. 4 ("at physiological pH LE
[L-Ergothioneine] does not auto-oxidize and is therefore very
stable in aqueous solution." . . . "remarkably stable to strong
alkali"); and Jinzhu Xu and Jean Claude Yadan. "Synthesis of
L-(+)-Ergothioneine", J. Org. Chem. 60:6296-6301, 1995 at p. 6296
("this explains its stability toward oxidative dimerization").
[0005] On the other hand, see U.S. Pat. No. 5,438,151, at column 1,
line 40 ("the very ready .beta.-elimination of the
trimethylammonium group in alkaline medium"); Donald Melville.
"Ergothioneine. Vitamins and Hormones", 17:155-204, 1959, at p. 161
("When Ergothioneine was boiled with 50% KOH solution,
trimethylamine was evolved and a yellow acid
C.sub.6H.sub.6O.sub.2N.sub.2S was formed"), and at p. 165 ("The two
most characteristic chemical reactions of Ergothioneine are the
ready oxidation of the sulfur atom and the lability of the
trimethylammonium radical toward alkali . . . the formation of
thiolurocanic acid and trimethylamine by the treatment of
Ergothioneine with hot, concentrated alkali has already been
considered.").
[0006] Ergothioneine and its solutions have no odor, despite the
presence of sulfur, because the sulfur is in the thione
conformation (C.dbd.S), which has no odor, rather than the
sulfhydryl conformation (C--SH), which has the odor of rotten eggs.
The literature does not mention the appearance of any odor during
storage under any conditions. In fact, European Patent No.
EP0483426 describes ergothioneine as one of the preferred
ingredients in a deodorant composition for topical application to
the skin. The contents of the documents cited herein are
incorporated herein by reference in their entirety.
[0007] The inventor has observed that a cosmetic formulation
containing 0.3% ergothioneine at pH 4.2-5.0, stored over several
months at room temperature, when rubbed onto a person's skin,
presented a fishy, amine odor to that person. Other cosmetic
formulations with more dilute solutions of 0.001% or 0.0005% at pH
7 also produced a fishy, amine odor when rubbed onto a person's
skin. However, in the cases of the more dilute solutions, the odor
was detected by some individuals and not others. The odor was
produced by stored samples which retained the same concentration of
the ergothioneine as they did initially.
[0008] The fishy, amine odor associated with ergothioneine is a
disadvantage in commercialization, particularly in oral
pharmaceuticals, injectibles pharmaceuticals, topical
pharmaceuticals, cosmetic products, nutritional supplements,
nutritional drinks, and other consumer products.
[0009] Those skilled in the art have found the removal or
prevention of the fishy, amine odor in aqueous solutions to be a
particularly difficult problem. U.S. Pat. No. 5,814,233 describes a
method for remediating methyl amine odors in aqueous systems using
a compound with amide or imide functionality, such as a hydantoin
composition, in the presence of hydrogen peroxide. The patent
further proposes reacting choline with hydantoin and hydrogen
peroxide. In U.S. Pat. Nos. 5,137,982 and 5,078,913, the inventors
describe a method for removing odor from a solution containing
trimethylamine and choline chloride using polybasic acid to react
with the choline chloride. The acids described include strong acids
such as sulfuric acid. U.S. Pat. No. 4,845,289 describes a method
for the removal or reduction of odor by use of methyl chloride at
temperatures above 50.degree. C. As disclosed in the patent, "In
these products as well as others, there frequently remains upon
completion of the reaction excess trimethylamine which is odiferous
and has undesirable toxicological problems. The residual
trimethylamine is difficult to remove completely from aqueous
system because of its high solubility in water, and extensive
purging with an inert gas and/or removal of water is necessary to
also remove substantial quantities of amine. This can be a
time-consuming and energy intensive process."
[0010] In addressing this concern, the initial assumption of the
inventor was that ergothioneine dimerizes by forming a disulfide
link between two molecules over time, which was not detected by the
analytical method. The inventor then reasoned that the dimer reacts
on the surface of a person's skin to release a compound that
produces an odor. The most widely recognized component of the human
body which can produce a fishy, amine smell is choline, which is a
component of prevalent skin lipids such as phosphatidyl choline.
Persons with large amounts of choline in their diets, or who are
defective in metabolizing choline, develop a fishy, amine odor on
their breaths and their bodies. It was therefore hypothesized by
the inventor that the reactant, perhaps choline, differed from one
person's skin to another, which would explain why some people
detected the odor after rubbing it on their skin and others did
not. In addition, it was theorized that the reaction required a few
seconds of heat from the skin, which would explain why the odor
appeared in a delayed fashion.
[0011] However, the inventor has now appreciated that the source of
the odor associated with the manufacture and storage of
ergothioneine is not related to the presence of choline, and that
none of the methods of the prior art are useful in preventing the
formation of the odor, produced after application to human skin, of
a product that contains ergothioneine but that does not contain
choline chloride.
[0012] The present invention describes practical methods which have
now been discovered to prevent the formation of the odor and/or
eliminate the odor, and to thereby remove this hindrance to
commercialization.
[0013] Without being limited to one particular theory, it is
believed that the odor associated with ergothioneine may be caused
by the formation of minute quantities of trimethylamine in solution
at neutral or even acidic pH over time. Trimethylamine is a gas
which has a strong ammonium odor but in trace amounts has a fishy,
amine odor. It is produced by rotting fish and other decaying
foods. The human nose is sensitive to minute quantities of
trimethylamine (odor threshold 25 parts per billion), which
probably evolved as a defense mechanism to detect contaminated
foods and thus to avoid illness. The gas is formed within the
aqueous phase of ergothioneine mixtures and remains dissolved, thus
masking the odor when the mixture is smelled directly. However,
when the ergothioneine mixture is applied to skin, the aqueous
components of the mixture are absorbed and the heat of the skin
volatilizes the trimethylamine. This may explain why, after rubbing
the mixture onto skin, the odor is detectable only after a few
seconds. Therefore, it is theorized that the variation in
perception of the odor by people is not a result of differences in
their skins or choline content but rather because of the
significant differences in olfactory sensitivity in the human
population, i.e. the ability to detect trimethylamine in trace
amounts. In fact, 5-7% of the human population is unable to smell
it at all, a condition called anosmia. HPLC analyses of the
ergothioneine concentrations did not detect the reduction in
ergothioneine or the formation of trimethylamine because the
changes were in trace amounts below the limits of detection of the
HPLC, but not below the limit of the human nose to detect
trimethylamine.
[0014] This observation by the inventor was confirmed by
reproducing the fishy, amine odor by boiling 2 mM (0.046%)
ergothioneine with 0.1 N NaOH (pH 11.5) for 10 minutes. The odor
was not ammonia-like, and was detectable by all persons. This
became the assay system used to test agents and procedures for
reduction or prevention of the odor.
[0015] It now has been discovered by the inventor that certain
trimethylamine absorbers, which also may be referred to as binders
or conjugators, are able both to prevent the formation of
trimethylamine from ergothioneine and to bind to trimethylamine
after its formation to prevent it from escaping as a gas detectable
by the nose.
[0016] In one embodiment of the present invention, the
trimethylamine binder is an acid, especially a weak acid.
[0017] In a further embodiment of the present invention, the
trimethylamine binder is sulfur dioxide or a compound which
generates sulfur dioxide upon its dissolution in water (i.e. sulfur
dioxide donors). The sulfur dioxide is able to bind trimethylamine
after its formation to prevent it from escaping as a gas detectable
by the nose. Thus, sulfur dioxide can function like the conjugated
base of a weak acid to prevent or reduce the odor of ergothioneine
during storage.
SUMMARY OF THE INVENTION
[0018] In accordance with a first aspect of the present invention,
there is provided an aqueous-containing treatment composition
comprising ergothioneine wherein the improvement is supplying the
ergothioneine to the treatment composition in the form of a
pre-blend comprising ergothioneine and at least one trimethylamine
absorber in an amount sufficient to ameliorate any trimethylamine
odor in the treatment composition when it is used for its intended
purpose.
[0019] In accordance with a further aspect of the invention, there
is provided a preblend composition consisting essentially of
ergothioneine at a concentration of greater than 2 mM (w/v) and a
trimethylamine absorber.
[0020] In accordance with yet a further aspect of the invention,
methods are provided for ameliorating a fishy, amine odor, due to
trimethylamine, formed in aqueous solutions, associated with the
processing or storage of a preparation containing ergothioneine.
Contemplated is a method comprising combining with the
ergothioneine, during the processing and/or prior to the storage of
the aqueous containing preparation, a trimethylamine absorber in an
amount sufficient to ameliorate the detection of the fishy, amine
odor in the composition when it used for its intended purpose. Also
contemplated is a method comprising introducing into an aqueous
containing preparation, after development of the odor, a
trimethylamine absorber in an amount sufficient to ameliorate the
detection of a fishy, amine odor, due to trimethylamine, by the
human nose. Further methods for ameliorating the trimethylamine
odor include a method of maintaining an aqueous-containing
ergothioneine-containing preparation at a pH of 7 or less, and a
method of maintaining the aqueous-containing
ergothioneine-containing preparation at a temperature of 25.degree.
C. or less.
BRIEF DESCRIPTION OF THE DRAWING
[0021] The FIGURE is a schematic drawing of the chemical structure
of ergothioneine.
DETAILED DESCRIPTION OF THE INVENTION AND ITS PREFERRED
EMBODIMENTS
[0022] As described in detail below, in certain of its preferred
aspects, the invention provides ergothioneine and
ergothioneine-containing mixtures which retain an innocuous odor
during preparation and/or storage, and methods for preventing the
formation of a fishy, amine odor, due to trimethylamine in
ergothioneine and ergothioneine-containing mixtures, or for
reducing, minimizing or substantially eliminating the fishy, amine
odor, due to trimethylamine, from ergothioneine or
ergothioneine-containing mixtures after formation of the odor.
[0023] All expressions of percent concentration of ergothioneine
herein mean percent weight by volume, unless otherwise explicitly
stated.
[0024] There are several methods for reducing, minimizing or
preventing the formation of the fishy, amine odor from an
ergothioneine containing mixtures. The odor can be minimized by
reducing levels of ergothioneine. A solution containing
ergothioneine in an aqueous medium (or in a non-aqueous polar
solvent, such as ethanol, butylene glycol, ethylene glycol,
ethylene glycol, isopropanol, or similar monohydric alcohol) at
less than about 20 .mu.M (0.00046%), for example, a solution at
0.00001% or less does not produce the characteristic odor.
[0025] The formation of the trimethylamine odor also can be
minimized during manufacturing of ergothioneine or ergothioneine
containing mixtures by maintaining at all times an acidic pH,
specifically a pH equal to or less than 7.0, during manufacturing
and in the final product. The lower the pH, the less likely is the
formation of the odor. A condition of alkalinity, meaning a pH
greater than 7.0, even for a brief time, is to be avoided.
[0026] In another method, the formation of the trimethylamine odor
can be minimized by avoiding heating ergothioneine or the
ergothioneine containing mixture above ambient temperature,
specifically 25.degree. C., even for brief periods of time. This is
particularly the case if sodium hydroxide or other strong or weak
base is contained in or added to the ergothioneine containing
mixture, or if the mixture is alkaline, meaning pH.gtoreq.8.0. For
long-term storage of ergothioneine or ergothioneine containing
mixtures, the odor can be minimized or prevented by maintaining the
mixture at or below ambient temperatures, specifically at
refrigerated temperatures of about 2-8.degree. C. or at freezing
temperatures of about -20.degree. C. to -70.degree. C. In this
case, ergothioneine is added in the last step, or in a step near
the end, of the manufacturing procedure, after all other components
have been added, the mixture is cool and the pH has been
appropriately adjusted.
[0027] The trimethylamine odor can be minimized during
manufacturing by avoiding adding ergothioneine or an
ergothioneine-containing mixture to a mixture with sodium hydroxide
or other strong or weak base, even if the resulting mixture is
neutralized soon after.
[0028] The formation of the fishy, amine odor, due to
trimethylamine, can be minimized or eliminated by adding to the
ergothioneine or the ergothioneine-containing mixture a
trimethylamine binder or absorber, more particularly, an acid. The
preferred acid is a weak acid. A weak acid is an acid that does not
ionize in solution to a significant extent. The K.sub.a (acid
dissociation constant, equal to the product of the concentrations
of the hydronium ion and the conjugated base divided by the
concentration of the undissociated acid) for a weak acid is between
1.8.times.10.sup.-16 and 55.5. If an acid can release more than one
hydronium ion, a separate K.sub.a is calculated for each hydronium
ion. Acids with K.sub.a greater than 55.5 are strong acids and
almost totally dissociate in water. A specifically preferred acid
is one having a K.sub.a between 1.8.times.10.sup.-16 and
5.5.times.10.sup.-1, more preferably, an acid having a K.sub.a
between 1.82.times.10.sup.-1 and 1.62.times.10.sup.-12. Most
preferred is citric acid. However, other weak acids useful in the
present invention include, but are not limited to, other
alpha-hydroxy acids, such as, lactic, glycolic, malic and tartaric
acids; carboxylic acids, for example, acetic, butanoic, formic,
heptanoic, hexanoic, octanoic, oxalic, pentanoic, propanoic acids;
other organic acids, for example, sugar acids, such as, ascorbic
acid, and purines, for example, uric acid; inorganic acids, such as
boric, carbonic, chromic, hydrocyanic, hydrofluoric, nitrous,
phosphoric acid, sulfuric and sulfurous acids, can be used. The
amount of the acid used should be sufficient to prevent the
detection of the fishy, trimethylamine odor, and this amount for a
specific weak acid can be determined by one ordinarily skilled in
the art. The ratio of the weight of the acid to the weight of
ergothioneine may be in the range of from about 1:1 to about 6:1.
Preferably the ratio is greater than about 1:1, such as from about
2:1 to about 4:1, and more preferably, the ratio is about 4:1,
respectively.
[0029] A particular category of compounds that is useful in
preventing, reducing or minimizing the formation of the fishy,
trimethylamine odor includes sulfur dioxide and sulfur dioxide
donors; that is, compounds that release sulfur dioxide upon
dissociation. Examples of such compounds are sodium sulfite, sodium
bisulfate, sodium metabisulfite, potassium bisulfate or potassium
metabisulfite, and calcium bisulfate. Sulfur dioxide gas may be
used directly. In this case the pre-blend of ergothioneine and
sulfur dioxide or sulfur dioxide donor compound may comprise from
about 0.000001 to about 45%, preferably from about 0.00001 to about
30%, more preferably from about 0.0001 to about 20% ergothioneine,
and from about 0.000001 to about 90%, preferably from about 0.00001
to about 60%, and more preferably from about 0.0001 to about 40% of
the trimethylamine absorber, in solution, suspension, or emulsion
of an aqueous or non-aqueous polar solvent which may be water or
butylene glycol, propylene glycol, ethanol, propanol, isopropanol,
and so on. In one preferred embodiment, a ratio of about 1:4
ergothioneine:sodium metabisulfite is most preferred.
[0030] While the acids, including weak acids, and sulfur dioxide or
sulfur dioxide releasing compounds may be used to prevent or
minimize the formation of the fishy, trimethylamine odor, they may
also be used to minimize or eliminate the odor after its formation.
This is believed to be due, in part or in whole, to the ability of
the weak acid and/or sulfur dioxide to bind trimethylamine gas and
retain it within the ergothioneine containing mixture.
[0031] In general, the weaker the acid the stronger it binds
trimethylamine, and therefore the better it is at suppressing the
fishy, trimethylamine odor. The weakness of an acid is measured by
its K.sub.a (acidity constant). The smaller the K.sub.a the weaker
the acid. A table of some common acids and their K.sub.a for each
of their ionized forms is shown in Table 1. The indicia "E-x", with
x referring to a number, means exponent. For example, with respect
to acetic acid, 1.74 E-5 means 1.74.times.10.sup.-5.
TABLE-US-00001 TABLE 1 Acids and their K.sub.a values ACID FORMULA
K.sub.a acetic acid H(C2H3O2) 1.74E-5 ascorbic acid (1) H2(C6H6O6)
7.94E-5 ascorbic acid (2) (HC6H6O6).sup.- 1.62E-12 boric acid (1)
H3BO3 5.37E-10 boric acid (2) (H2BO3).sup.- 1.8E-13 boric acid (3)
(HBO3).sup.= 1.6E-14 butanoic acid H(C4H7O2) 1.48E-5 carbonic acid
(1) H2CO3 4.47E-7 carbonic acid (2) (HCO3).sup.- 4.68E-11 chromic
acid (1) H2CrO4 1.82E-1 chromic acid (2) (HCrO4).sup.- 3.24E-7
citric acid (1) H3(C6H5O7) 7.24E-4 citric acid (2) (H2C6H5O7).sup.-
1.70E-5 citric acid (3) (HC6H5O7).sup.= 4.07E-7 formic acid H(CHO2)
1.78E-4 heptanoic acid H(C7H13O2) 1.29E-5 hexanoic acid H(C6H11O2)
1.41E-5 hydrocyanic acid HCN 6.17E-10 hydrofluoric acid HF 6.31E-4
lactic acid H(C3H5O3) 8.32E-4 nitrous acid HNO2 5.62E-4 octanoic
acid H(C8H15O2) 1.29E-4 oxalic acid (1) H2(C204) 5.89E-2 oxalic
acid (2) (HC2O4).sup.- 6.46E-5 pentanoic acid H(C5H9O2) 3.31E-5
phosphoric acid (1) H3PO4 6.92E-3 phosphoric acid (2) (H2PO4).sup.-
6.17E-8 phosphoric acid (3) (HPO4).sup.= 2.09E-12 propanoic acid
H(C3H5O2) 1.38E-5 sulfuric acid (2) (HSO4)- 1.05E-2 sulfurous acid
(1) H2SO3 1.41E-2 sulfurous acid (2) (HSO3).sup.- 6.31E-8 uric acid
H(C5H3N4O3) 1.29E-4
[0032] Other factors which may be considered, for purposes of
commercialization, in the selection of a compound, particularly the
weak acid, that meets the requirements of this invention, include
the pH of the mixture at the concentration of the weak acid
required to minimize, prevent or eliminate the odor, the
compatibility of the compound with other components of the mixture,
side effects of the compound, regulations of the government or
other authorities controlling the compound or its use, the cost of
the compound, side reactions of the compound with other components
of the mixture or with parts of the human body, or the taste, smell
or feel of the compound. For example, oral products containing
sulfites, such as sodium metabisulfite, produce allergic reactions
in some persons, and therefore in some stocks, intermediate
products or finished products this may not be the most favorable
compound of the invention. As another example, sodium dioxide is a
highly reactive gas and has a characteristic odor, and therefore in
some circumstances may not be the preferable compound for use in
minimizing, prevention or eliminating the fishy, trimethylamine
odor.
[0033] The invention can be used with ergothioneine or
ergothioneine-containing mixtures such as pure ergothioneine, or a
pre-blend comprising ergothioneine in an aqueous or organic
solution or emulsion that results from manufacturing, or that
constitute the form of shipment or transfer of ergothioneine from
one place to another. The pre-blend preferably comprises from about
0.000001 to about 45%, preferably from about 0.00001 to about 30%,
more preferably from about 0.0001 to about 20% ergothioneine, and
from about 0.000001 to about 90%, preferably from about 0.00001 to
about 60%, and more preferably from about 0.0001 to about 40% of
the trimethylamine absorber, in solution, suspension, or emulsion
with a polar solvent which may be a polar aqueous solvent such as
water, or a mixture of water and non-aqueous polar solvents such as
C.sub.1-4 monohydric alcohols, or C.sub.1-4 dihydric alcohols (such
as ethanol, propanol, isopropanol, glycerol, butylene glycol,
propylene glycol) where in said polar solvent is present in an
amount ranging from about 10 to about 99.99999%, with all
percentages mentioned herein being percentages by weight unless
otherwise indicated. The pre-blend may contain other ingredients
such as preservatives, antioxidants, or other ingredients which
enhance stability or provide other commercially beneficial effects.
The ergothioneine-containing composition may be sold in the form of
the pre-blend to manufacturers of cosmetic, pharmaceutical, or OTC
products for their use in formulating such products. The presence
of the trimethylamine absorber in the pre-blend ameliorates any
unpleasant odor in the pre-blend as well as when it is formulated
into cosmetic or pharmaceutical products or otherwise used for its
intended purpose.
[0034] The invention can also be used in preparation of
ergothioneine in aqueous or organic solution or emulsion in
products as finished goods. These products can include skincare
products, consumer products, over the counter products, oral
supplements, nutritional supplements, food, seasoning for food, or
pharmaceutical drugs. The products can be used by topical
application, subcutaneous injection, intramuscular injection,
intravenous injection or any other injection, or by oral or nasal
ingestion, or by eye or ear drops, or by suppositories. The
nutritional supplements can be for oral ingestion including pills,
juices, shakes, power drinks, fortified foods and fortified or
supplemented water.
[0035] The ergothioneine may be in a pure form before making use of
this invention. The ergothioneine may also be in the form of a
pre-blend or in a mixture with other compounds. Most preferred is
where the ergothioneine will be present in the pre-blend
compositions in the amount of from equal to or greater than about 2
mM (0.046% w/v), for example, from about 2 mM to about 70 mM,
preferably equal to or greater than about 3 mM, for example, from
about 3 mM to about 50 mM, and more preferably, equal to or greater
than about 4 mM, for example from about 4 mM to about 30 mM.
Compositions into which ergothioneine may be incorporated may be
emulsions, solutions, suspensions, gels, or anhydrous compositions.
If emulsions, they may be water-in-oil or oil-in-water, comprising
from about 0.1 to 95%, preferably from about 0.5 to about 90%, and
more preferably from about 1 to 85% water, by weight of the total
composition. If in the form of aqueous solutions, suspensions or
gels, the composition may contain from about 10 to about 99% water
with the remaining ingredients being one or more actives. The
ergothioneine or the pre-blend may be solubilized or dispersed in
either the aqueous phase or the oil phase of the emulsion. In one
preferred embodiment, the ergothioneine is solubilized or dispersed
in the aqueous phase. The compositions of the present invention may
be in a non-aqueous or anhydrous form in which the ergothioneine is
solubilized or dispersed in a polar non-aqueous solvent, such as
ethanol, propylene glycol, butylene glycol, or non-polar oils, and
the like.
[0036] In the event the compositions of the invention into which
the ergothioneine is incorporated, either neat or in the form of
the pre-blend, are in emulsion form, the composition will comprise
an oil phase. Oily ingredients are desirable for the skin
moisturizing and protective properties. Suitable oils include
silicones, esters, vegetable oils, including but not limited to
those set forth herein. The oils may be volatile or nonvolatile,
and are preferably in the form of a pourable liquid at room
temperature. The term "volatile" means that the oil has a
measurable vapor pressure or a vapor pressure of at least about 2
mm. of mercury at 20.degree. C. The term "nonvolatile" means that
the oil has a vapor pressure of less than about 2 mm. of mercury at
20.degree. C.
[0037] Cyclic silicones are one type of volatile silicone that may
be used in the composition. Such silicones have the general
formula:
##STR00001##
where n=3-6, preferably 4, 5, or 6.
[0038] Also suitable are linear volatile silicones, for example,
those having the general formula:
(CH.sub.3).sub.3--Si--O--[Si--(CH.sub.3).sub.2--O].sub.n--Si(CH.sub.3).s-
ub.3
[0039] where n=0, 1, 2, 3, 4, or 5, preferably 0, 1, 2, 3, or
4.
[0040] Cyclic and linear volatile silicones are available from
various commercial sources including Dow Corning Corporation and
General Electric. The Dow Corning linear volatile silicones are
sold under the tradenames Dow Corning 244, 245, 344, 345 and 200
fluids. These fluids include hexamethyldisiloxane (viscosity 0.65
centistokes (abbreviated cst)), octamethyltrisiloxane (1.0 cst),
decamethyltetrasiloxane (1.5 cst), dodecamethylpentasiloxane (2
cst) and mixtures thereof, with all viscosity measurements being at
25.degree. C. A preferred cyclic volatile silicone is
cyclopentasiloxane, available from Dow Corning as DC 345 Fluid.
[0041] Suitable branched volatile silicones include alkyl
trimethicones such as methyl trimethicone, a branched volatile
silicone having the general formula:
##STR00002##
[0042] Methyl trimethicone may be purchased from Shin-Etsu
Silicones under the tradename TMF-1.5, having a viscosity of 1.5
cst at 25.degree. C.
[0043] Nonvolatile silicone oils, both water soluble and water
insoluble, are also suitable for use in the composition. Such
silicones preferably have a viscosity ranging from about greater
than 5 to 800,000 cst, preferably 20 to 200,000 cst at 25.degree.
C. Suitable water insoluble silicones include amine functional
silicones such as amodimethicone.
[0044] For example, such nonvolatile silicones may have the
following general formula:
##STR00003##
wherein R and R' are each independently C.sub.1-30 straight or
branched chain, saturated or unsaturated alkyl, phenyl or aryl,
trialkylsiloxy, and x and y are each independently 1-1,000,000;
with the proviso that there is at least one of either x or y, and A
is alkyl siloxy endcap unit. Preferred is where A is a methyl
siloxy endcap unit; in particular trimethylsiloxy, and R and R' are
each independently a C.sub.1-30 straight or branched chain alkyl,
phenyl, or trimethylsiloxy, more preferably a C.sub.1-22 alkyl,
phenyl, or trimethylsiloxy, most preferably methyl, phenyl, or
trimethylsiloxy, and resulting silicone is dimethicone, phenyl
dimethicone, diphenyl dimethicone, phenyl trimethicone, or
trimethylsiloxyphenyl dimethicone. Other examples include alkyl
dimethicones such as cetyl dimethicone, and the like wherein at
least one R is a fatty alkyl (C.sub.12, C.sub.14, C.sub.16,
C.sub.18, C.sub.20, or C.sub.22), and the other R is methyl, and A
is a trimethylsiloxy endcap unit, provided such alkyl dimethicone
is a pourable liquid at room temperature. Preferred is dimethicone
which can be purchased from Dow Corning Corporation as DC 200/100
cs fluid.
[0045] A variety of nonvolatile oils are also suitable for use in
the compositions of the invention. The nonvolatile oils generally
have a viscosity of greater than about 5 to 10 centistokes at
25.degree. C., and may range in viscosity up to about 1,000,000
centipoise at 25.degree. C. Examples of nonvolatile oils include,
but are not limited to esters and hydrocarbon oils.
[0046] Suitable esters are mono-, di-, and triesters. The
composition may comprise one or more esters selected from the
group, or mixtures thereof.
[0047] Monoesters are defined as esters formed by the reaction of a
monocarboxylic acid having the formula R--COOH, wherein R is a
straight or branched chain saturated or unsaturated alkyl having 2
to 45 carbon atoms, or phenyl; and an alcohol having the formula
R--OH wherein R is a straight or branched chain saturated or
unsaturated alkyl having 2-30 carbon atoms, or phenyl. Both the
alcohol and the acid may be substituted with one or more hydroxyl
groups. Either one or both of the acid or alcohol may be a "fatty"
acid or alcohol, and may have from about 6 to 30 carbon atoms, more
preferably 12, 14, 16, 18, or 22 carbon atoms in straight or
branched chain, saturated or unsaturated form. Examples of
monoester oils that may be used in the compositions of the
invention include hexyl laurate, butyl isostearate, hexadecyl
isostearate, cetyl palmitate, isostearyl neopentanoate, stearyl
heptanoate, isostearyl isononanoate, stearyl lactate, stearyl
octanoate, stearyl stearate, isononyl isononanoate, and so on.
[0048] Suitable diesters are the reaction product of a dicarboxylic
acid and an aliphatic or aromatic alcohol or an aliphatic or
aromatic alcohol having at least two substituted hydroxyl groups
and a monocarboxylic acid. The dicarboxylic acid may contain from 2
to 30 carbon atoms, and may be in the straight or branched chain,
saturated or unsaturated form. The dicarboxylic acid may be
substituted with one or more hydroxyl groups. The aliphatic or
aromatic alcohol may also contain 2 to 30 carbon atoms, and may be
in the straight or branched chain, saturated, or unsaturated form.
Preferably, one or more of the acid or alcohol is a fatty acid or
alcohol, i.e. contains 12-22 carbon atoms. The dicarboxylic acid
may also be an alpha hydroxy acid. The ester may be in the dimer or
trimer form. Examples of diester oils that may be used in the
compositions of the invention include diisotearyl malate, neopentyl
glycol dioctanoate, dibutyl sebacate, dicetearyl dimer dilinoleate,
dicetyl adipate, diisocetyl adipate, diisononyl adipate,
diisostearyl dimer dilinoleate, diisostearyl fumarate, diisostearyl
malate, dioctyl malate, and so on.
[0049] Suitable triesters comprise the reaction product of a
tricarboxylic acid and an aliphatic or aromatic alcohol or
alternatively the reaction product of an aliphatic or aromatic
alcohol having three or more substituted hydroxyl groups with a
monocarboxylic acid. As with the mono- and diesters mentioned
above, the acid and alcohol contain 2 to 30 carbon atoms, and may
be saturated or unsaturated, straight or branched chain, and may be
substituted with one or more hydroxyl groups. Preferably, one or
more of the acid or alcohol is a fatty acid or alcohol containing
12 to 22 carbon atoms. Examples of triesters include esters of
arachidonic, citric, or behenic acids, such as triarachidin,
tributyl citrate, triisostearyl citrate, tri C.sub.12-13 alkyl
citrate, tricaprylin, tricaprylyl citrate, tridecyl behenate,
trioctyldodecyl citrate, tridecyl behenate; or tridecyl cocoate,
tridecyl isononanoate, and so on.
[0050] Esters suitable for use in the composition are further
described in the C.T.F.A. Cosmetic Ingredient Dictionary and
Handbook, Eleventh Edition, 2006, under the classification of
"Esters", the text of which is hereby incorporated by reference in
its entirety.
[0051] It may be desirable to incorporate one or more nonvolatile
hydrocarbon oils into the composition. Suitable nonvolatile
hydrocarbon oils include paraffinic hydrocarbons and olefins,
preferably those having greater than about 20 carbon atoms.
Examples of such hydrocarbon oils include C.sub.24-28 olefins,
C.sub.30-45 olefins, C.sub.20-40 isoparaffins, hydrogenated
polyisobutene, polyisobutene, polydecene, hydrogenated polydecene,
mineral oil, pentahydrosqualene, squalene, squalane, and mixtures
thereof. In one preferred embodiment such hydrocarbons have a
molecular weight ranging from about 300 to 1000 Daltons.
[0052] Surface active agents which may be used in the compositions
of the invention include silicone surfactants and organic nonionic
surfactants. If used, surface active agents are present in the
range of from about 0.1 to about 80%, preferably in the range of
from about 1 to 50%, and more preferably in the range of from about
5 to about 40%, based on the total weight of the composition.
[0053] Suitable silicone surfactants include polyorganosiloxane
polymers that have amphiphilic properties, for example contain
hydrophilic radicals and lipophilic radicals. These silicone
surfactants may be liquids or solids at room temperature.
[0054] One type of silicone surfactant that may be used is
generally referred to as dimethicone copolyol or alkyl dimethicone
copolyol. This surfactant is either a water-in-oil or oil-in-water
surfactant having a Hydrophile/Lipophile Balance (HLB) ranging from
about 2 to 18. Preferably the silicone surfactant is a nonionic
surfactant having an HLB ranging from about 2 to 12, preferably
about 2 to 10, most preferably about 4 to 6. The term "hydrophilic
radical" means a radical that, when substituted onto the
organosiloxane polymer backbone, confers hydrophilic properties to
the substituted portion of the polymer. Examples of radicals that
will confer hydrophilicity are hydroxy-polyethyleneoxy, hydroxyl,
carboxylates, and mixtures thereof. The term "lipophilic radical"
means an organic radical that, when substituted onto the
organosiloxane polymer backbone, confers lipophilic properties to
the substituted portion of the polymer. Examples of organic
radicals that will confer lipophilicity are C.sub.1-40 straight or
branched chain alkyl, fluoro, aryl, aryloxy, C.sub.1-40 hydrocarbyl
acyl, hydroxy-polypropyleneoxy, or mixtures thereof.
[0055] One type of suitable silicone surfactant has the general
formula:
##STR00004##
wherein p is 0-40 (the range including all numbers between and
subranges such as 2, 3, 4, 13, 14, 15, 16, 17, 18, etc.), and PE is
(--C.sub.2H.sub.4O).sub.a--(--C.sub.3H.sub.6O).sub.b--H wherein a
is 0 to 25, b is 0-25 with the proviso that both a and b cannot be
0 simultaneously, x and y are each independently ranging from 0 to
1 million with the proviso that they both cannot be 0
simultaneously. In one preferred embodiment, x, y, z, a, and b are
such that the molecular weight of the polymer ranges from about
5,000 to about 500,000, more preferably from about 10,000 to
100,000, and is most preferably approximately about 50,000 and the
polymer is generically referred to as dimethicone copolyol. One
type of silicone surfactant is wherein p is such that the long
chain alkyl is cetyl or lauryl, and the surfactant is called,
generically, cetyl dimethicone copolyol or lauryl dimethicone
copolyol respectively.
[0056] In some cases the number of repeating ethylene oxide or
propylene oxide units in the polymer are also specified, such as a
dimethicone copolyol that is also referred to as PEG-15/PPG-10
dimethicone, which refers to a dimethicone having substituents
containing 15 ethylene glycol units and 10 propylene glycol units
on the siloxane backbone. It is also possible for one or more of
the methyl groups in the above general structure to be substituted
with a longer chain alkyl (e.g. ethyl, propyl, butyl, etc.) or an
ether such as methyl ether, ethyl ether, propyl ether, butyl ether,
and the like.
[0057] Examples of silicone surfactants are those sold by Dow
Corning under the tradename Dow Corning 3225C Formulation Aid
having the CTFA name cyclotetrasiloxane (and) cyclopentasiloxane
(and) PEG/PPG-18 dimethicone; or 5225C Formulation Aid, having the
CTFA name cyclopentasiloxane (and) PEG/PPG-18/18 dimethicone; or
Dow Corning 190 Surfactant having the CTFA name PEG/PPG-18/18
dimethicone; or Dow Corning 193 Fluid, Dow Corning 5200 having the
CTFA name lauryl PEG/PPG-18/18 methicone; or Abil EM 90 having the
CTFA name cetyl PEG/PPG-14/14 dimethicone sold by Goldschmidt; or
Abil EM 97 having the CTFA name bis-cetyl PEG/PPG-14/14 dimethicone
sold by Goldschmidt; or Abil WE 09 having the CTFA name cetyl
PEG/PPG-10/1 dimethicone in a mixture also containing
polyglyceryl-4 isostearate and hexyl laurate; or KF-6011 sold by
Shin-Etsu Silicones having the CTFA name PEG-11 methyl ether
dimethicone; KF-6012 sold by Shin-Etsu Silicones having the CTFA
name PEG/PPG-20/22 butyl ether dimethicone; or KF-6013 sold by
Shin-Etsu Silicones having the CTFA name PEG-9 dimethicone; or
KF-6015 sold by Shin-Etsu Silicones having the CTFA name PEG-3
dimethicone; or KF-6016 sold by Shin-Etsu Silicones having the CTFA
name PEG-9 methyl ether dimethicone; or KF-6017 sold by Shin-Etsu
Silicones having the CTFA name PEG-10 dimethicone; or KF-6038 sold
by Shin-Etsu Silicones having the CTFA name lauryl PEG-9
polydimethylsiloxyethyl dimethicone.
[0058] Also suitable are various types of crosslinked silicone
surfactants that are often referred to as emulsifying elastomers.
They are typically prepared as set forth above with respect to the
section "silicone elastomers" except that the silicone elastomers
will contain at least one hydrophilic moiety such as
polyoxyalkylenated groups. Typically these polyoxyalkylenated
silicone elastomers are crosslinked organopolysiloxanes that may be
obtained by a crosslinking addition reaction of
diorganopolysiloxane comprising at least one hydrogen bonded to
silicon and of a polyoxyalkylene comprising at least two
ethylenically unsaturated groups. In at least one embodiment, the
polyoxyalkylenated crosslinked organo-polysiloxanes are obtained by
a crosslinking addition reaction of a diorganopolysiloxane
comprising at least two hydrogens each bonded to a silicon, and a
polyoxyalkylene comprising at least two ethylenically unsaturated
groups, optionally in the presence of a platinum catalyst, as
described, for example, in U.S. Pat. No. 5,236,986 and U.S. Pat.
No. 5,412,004, U.S. Pat. No. 5,837,793 and U.S. Pat. No. 5,811,487,
the contents of which are incorporated by reference.
[0059] Polyoxyalkylenated silicone elastomers that may be used in
at least one embodiment of the invention include those sold by
Shin-Etsu Silicones under the names KSG-21, KSG-20, KSG-30, KSG-31,
KSG-32, KSG-33; KSG-210 which is dimethicone/PEG-10/15 crosspolymer
dispersed in dimethicone; KSG-310 which is PEG-15 lauryl
dimethicone crosspolymer; KSG-320 which is PEG-15 lauryl
dimethicone crosspolymer dispersed in isododecane; KSG-330 (the
former dispersed in triethylhexanoin), KSG-340 which is a mixture
of PEG-10 lauryl dimethicone crosspolymer and PEG-15 lauryl
dimethicone crosspolymer.
[0060] Also suitable are polyglycerolated silicone elastomers like
those disclosed in PCT/WO 2004/024798, which is hereby incorporated
by reference in its entirety. Such elastomers include Shin-Etsu's
KSG series, such as KSG-710 which is dimethicone/polyglycerin-3
crosspolymer dispersed in dimethicone; or lauryl
dimethicone/polyglycerin-3 crosspolymer dispersed in a variety of
solvent such as isododecane, dimethicone, triethylhexanoin, sold
under the Shin-Etsu tradenames KSG-810, KSG-820, KSG-830, or
KSG-840. Also suitable are silicones sold by Dow Corning under the
tradenames 9010 and DC9011.
[0061] One preferred crosslinked silicone elastomer emulsifier is
dimethicone/PEG-10/15 crosspolymer, which provides excellent
aesthetics due to its elastomeric backbone, but also surfactancy
properties.
[0062] The composition may comprise one or more nonionic organic
surfactants. Suitable nonionic surfactants include alkoxylated
alcohols, or ethers, formed by the reaction of an alcohol with an
alkylene oxide, usually ethylene or propylene oxide. Preferably the
alcohol is either a fatty alcohol having 6 to 30 carbon atoms.
Examples of such ingredients include Steareth 2-100, which is
formed by the reaction of stearyl alcohol and ethylene oxide and
the number of ethylene oxide units ranges from 2 to 100; Beheneth
5-30 which is formed by the reaction of behenyl alcohol and
ethylene oxide where the number of repeating ethylene oxide units
is 5 to 30; Ceteareth 2-100, formed by the reaction of a mixture of
cetyl and stearyl alcohol with ethylene oxide, where the number of
repeating ethylene oxide units in the molecule is 2 to 100; Ceteth
1-45 which is formed by the reaction of cetyl alcohol and ethylene
oxide, and the number of repeating ethylene oxide units is 1 to 45,
and so on.
[0063] Other alkoxylated alcohols are formed by the reaction of
fatty acids and mono-, di- or polyhydric alcohols with an alkylene
oxide. For example, the reaction products of C.sub.6-30 fatty
carboxylic acids and polyhydric alcohols which are monosaccharides
such as glucose, galactose, methyl glucose, and the like, with an
alkoxylated alcohol. Examples include polymeric alkylene glycols
reacted with glyceryl fatty acid esters such as PEG glyceryl
oleates, PEG glyceryl stearate; or PEG polyhydroxyalkanotes such as
PEG dipolyhydroxystearate wherein the number of repeating ethylene
glycol units ranges from 3 to 1000.
[0064] Also suitable as nonionic surfactants are those formed by
the reaction of a carboxylic acid with an alkylene oxide or with a
polymeric ether. The resulting products have the general
formula:
##STR00005##
where RCO is the carboxylic ester radical, X is hydrogen or lower
alkyl, and n is the number of polymerized alkoxy groups. In the
case of the diesters, the two RCO-- groups do not need to be
identical. Preferably, R is a C6-30 straight or branched chain,
saturated or unsaturated alkyl, and n is from 1-100.
[0065] Monomeric, homopolymeric, or block copolymeric ethers are
also suitable as nonionic surfactants. Typically, such ethers are
formed by the polymerization of monomeric alkylene oxides,
generally ethylene or propylene oxide. Such polymeric ethers have
the following general formula:
##STR00006##
wherein R is H or lower alkyl and n is the number of repeating
monomer units, and ranges from 1 to 500.
[0066] Other suitable nonionic surfactants include alkoxylated
sorbitan and alkoxylated sorbitan derivatives. For example,
alkoxylation, in particular ethoxylation of sorbitan provides
polyalkoxylated sorbitan derivatives. Esterification of
polyalkoxylated sorbitan provides sorbitan esters such as the
polysorbates. For example, the polyalkyoxylated sorbitan can be
esterified with C.sub.6-30, preferably C.sub.12-22 fatty acids.
Examples of such ingredients include Polysorbates 20-85, more
specifically Polysorbate 80, sorbitan oleate, sorbitan
sesquioleate, sorbitan palmitate, sorbitan sesquiisostearate,
sorbitan stearate, and so on. Specifically preferred for use in the
present invention is Polysorbate 80, available as Liposorb O-20
from Lipo Chemicals.
[0067] Other compounds which may be found in the compositions of
the present invention include, but are not limited to: buffers and
salts to adjust the pH of the solution; preservatives and
anti-microbial agents, such as Botanistat PF-64, available from D-D
Chemco, Inc.; agents that restore the antioxidant capacity of
ergothioneine such as forms of Vitamin C; other antioxidants, such
as DNA repair extracts encapsulated in liposomes, such as
Roxisomes.RTM. (Arabidposis Exact/lecithin/water/phenoxyethanol),
Ultrasomes.RTM. (Micrococcus lysate); vitamins such as vitamin A or
vitamin E; nutrients both essential and non-essential, such as
amino acids and minerals; compounds for brightening and lighting
skin such as undecylenoyl phenylalanine, available as Sepiwhite MSH
from Seppic, and mulberry extract; compounds for preventing and
reducing irritation and inflammation, such as Evodiox (Evodia
Rutaecapra Fruit Extract/butylene glycol/phenoxyethanol); compounds
for preventing or slowing aging; compounds that protect against
environmental insult and toxins such as ultraviolet light and
pollution; compounds for treating disease or cancer; and compounds
for preventing disease such as vaccines. It may also be desirable
to include one or more humectants in the composition. If present,
such humectants may range from about 0.001 to 25%, preferably from
about 0.005 to 20%, more preferably from about 0.1 to 15% by weight
of the total composition. Examples of suitable humectants include
glycols, sugars, and the like. Suitable glycols are in monomeric or
polymeric form and include polyethylene and polypropylene glycols
such as PEG 4-200, which are polyethylene glycols having from 4 to
200 repeating ethylene oxide units; as well as C.sub.1-6 alkylene
glycols such as propylene glycol, butylene glycol, pentylene
glycol, and the like. Suitable sugars, some of which are also
polyhydric alcohols, are also suitable humectants. Examples of such
sugars include glucose, fructose, honey, hydrogenated honey,
inositol, maltose, mannitol, maltitol, sorbitol, sucrose, xylitol,
xylose, and so on. Also suitable is urea. The humectants used in
the composition of the invention may be C.sub.1-6, preferably
C.sub.2-4 alkylene glycols, such as butylene glycol. A preferred
humectant used in the compositions of the invention is glycerin.
These other compounds will be present in the range of from about
0.0001 to about 40% by weight of the composition.
[0068] Mixtures of ergothioneine with compounds not listed here may
also make use of the invention.
[0069] Without intending to restrict in any way the scope of the
invention, the following examples are presented to illustrate the
invention's aspects and its use.
Example 1
Stability of Ergothioneine with and without Citric Acid
[0070] A sample of ergothioneine was prepared at 2 mM or 0.046%
(w/v) in water with or without 0.2% (w/v) citric acid and placed in
glass vials of approximately 2 ml volume, purged of air with
nitrogen gas and sealed. The samples were stored at 4.degree. C.,
25.degree. C. or 40.degree. C. for 30 or 60 days and then analyzed
by HPLC. Each vial was used once and then discarded. Each
measurement was made in triplicate and the results averaged. A
standard curve of ergothioneine concentration was constructed at
each time point to quantify the amount of ergothioneine in each
vial. Each amount in Table 2 is expressed as percentage of the
amount measured in the initial sample at the start of the
experiment.
TABLE-US-00002 TABLE 2 Stability of ergothioneine during storage
with and without citric acid as a percent of the starting
concentration % EGT without % EGT with Temperature Time citric acid
citric acid 4.degree. C. 30 days 96.1 101.6 4.degree. C. 60 days
106.5 105.8 25.degree. C. 30 days 84.0 101.5 25.degree. C. 60 days
100.5 102.2 40.degree. C. 30 days 86.3 101.1 40.degree. C. 60 days
99.8 99.6
[0071] While there was some variability in the measurement of
ergothioneine without citric acid at 30 days, the results from the
60 day tests indicate little or no loss of ergothioneine
(decomposition or disassociation) during the incubation at
temperatures up to 40.degree. C. and no effect of citric acid on
the change in ergothioneine concentration.
Example 2
Development of Fishy Amine Odor at Room Temperature Storage
[0072] Lotions containing from 20 .mu.M (0.00046%) to 13 mM (0.3%)
ergothioneine (w/v) developed a similar fishy, amine odor over time
stored at ambient room temperature. These odors were not
immediately detected by smelling the container, but were detected
after application to the skin and allowing a few seconds before
smelling. Another lotion surprisingly did not.
TABLE-US-00003 TABLE 3 Formula 1: Lotion PHASE INGREDIENT % (w/v) 1
Water 61.80 1 Butylene Glycol 2.00 1 Phenoxyethanol 0.50 1 Disodium
EDTA 0.10 1 Magnesium Aluminum Silicate 10.00 2 Simmondsia
Chinensis (Jojoba) Butter 4.00 2 Hydrogenated Polyisobutene 3.00 2
Neopentyl Glycol Dihptanoate 5.00 2 Cetearyl Alcohol/Ceteareth-20
01.50 2 Cetyl Alcohol 01.75 2 Stearyl Alcohol 00.70 2
Tetrahexyldecyl Ascorbate 00.50 2 Stearic Acid 00.85 2 Glyceryl
Stearate/PEG-100 Stearate 05.00 2 1,2 Hexanediol/Capryl Glycol
01.00 3 Water 02.00 3 Ergothioneine 00.30 Total 100.00
[0073] Procedure:
[0074] Combine phase 1 ingredients in main beaker and heat to
78-80.degree. C. with moderate speed propeller mixing. Combine
phase 2 ingredients and heat to 80.degree. C. and mix until clear
and uniform. Slowly add phase 2 to phase 1 with moderate speed
propeller mixing. Cool batch to 40.degree. C. Then add premixed
phase 3 ingredients to the batch. Cool to 25.degree. C. Batch has a
final pH of about 4.8-5.2, and a viscosity of about 180,000-230,000
cps. The preparation of this 0.3% ergothioneine in water and
magnesium aluminum silicate base lotion, included a manufacturing
step in which the ergothioneine phase was added to a 40.degree. C.
solution and then cooled to 25.degree. C. It had no odor
immediately after manufacturing but developed a noticeable fishy
trimethylamine odor after less than a year in storage at ambient
temperature.
TABLE-US-00004 TABLE 4 Formula 2: Lotion PHASE INGREDIENT % (w/v) 1
Water 61.45 1 Glycerin 5.00 1 Disodium EDTA 0.05 1 Phenoxyethanol
0.50 1 Carbomer 12.50 2 Steareth-21 0.50 2 Steareth-2 0.10 2
Glyceryl Stearate SE 0.50 2 Cetearyl alcohol/Polysorbate 60 1.00 2
Dimethicone 4.00 2 Cetyl Alcohol 2.50 2 Hydrogenated
Coco-glycerides 1.50 2 Santalum Album (Sandalwood) Wood Extract/
3.00 Phellodendrom Amurense Bark Extract/ Hordeum Distichon
(Barley) Extract 2 1,2 Hexanediol/Caprylyl Glycol 1.00 3 Sodium
Hydroxide 0.03 4 Polyacrylamide/C13-14 Isoparaffin/Laureth-7 2.00 5
Evodia Rutaecarpa Fruit Extract/Butylene 0.30 Glycol/Phenoxyethanol
5 Rosemary Extract/Lecithin/Water 0.30 5 Ergothioneine 1.00 5
Retinol/Polysorbate-20/Lecithin/Water 1.00 6 Sodium Hydroxide
(adjust pH to 7.0-7.5) qs Total 100.00
[0075] Procedure: Combine phase 1 ingredients and heat to
78-80.degree. C. with moderate speed propeller mixing. Mix until
carbomer has completely dispersed. Separately, heat phase 2
ingredients to 80.degree. C. and mix until clear and uniform. Add
phase 2 to phase 1 with medium speed propeller mixing. Add phase 3
to the batch with medium speed propeller mixing. Cool the batch. At
50.degree. C., add phase 4 to the batch and increase the mixing
speed as the batch begins to thicken. At 30-35.degree. C., add
phase 5 ingredients individually to the batch. Mix well between the
additions. Cool batch to 25.degree. C. and adjust pH to 7.0-7.5
using phase 6 ingredient. The final viscosity of the batch is about
1,125,000-1,375,000 cps. The preparation of this 0.00046%
ergothioneine product in water and hydrogel base lotion had no odor
immediately after manufacturing but developed a slight fishy,
trimethylamine odor after approximately one year in storage at
ambient temperature.
TABLE-US-00005 TABLE 5 Formula 3: Lotion PHASE INGREDIENT % (w/v) 1
Water 70.95 1 Glycerin 2.50 1 Disodium EDTA 0.10 1 1,2
Hexanediol/Caprylyl Glycol 1.00 1 Sodium Hydroxide 0.50 2 C20-22
Alkyl Phosphate/C20-22 Alcohols 3.00 2 Squalane 3.50 2 Cetyl
Alcohol 1.75 2 Linoleic Acid 0.25 2 Tocopherol 0.20 2
Tetrahexyldecyl Ascorbate 0.50 2 Hydrogenated Coco-Glycerides 1.25
2 Glycine Soja (Soybean) Oil 2.25 2 Isocetyl Stearate 5.25 3 Sodium
Acrylate/Acrylyloyldimethyl Taurate/ 1.00
Copolymer/Isohexadecane/Polysorbate 80 3 Cyclopentasiloxane 4.00 3
Phenoxyethanol 0.50 4 Ergothioneine 1.00 4 Arabidposis
Extract/Lecithin/Water/ 0.50 Phenoxyethanol Total 100.00
[0076] Procedure: Combine phase 1 ingredients and heat to
78-80.degree. C. with medium speed propeller mixing. Separately
combine phase 2 ingredients and heat to 80.degree. C. Mix until
clear and uniform. Add phase 2 to phase 1 with medium speed
propeller mixing. Cool batch. At 50.degree. C., add phase 3
ingredients to batch, one at a time, mixing well after each
addition. Increase mixing speed as batch begins to thicken. At
25-30.degree. C., add phase 4 ingredients to the batch. Cool batch
to 25.degree. C. The final pH of the batch is about 4.6-5.0. The
viscosity is about 13,500-16,500 cps. The preparation of this
0.00046% ergothioneine in a water, glycerin and silicone base
lotion included a manufacturing step in which ergothioneine was
added at the end of the process at 25.degree. C. It developed a
very faint fishy trimethylamine odor after approximately one year
in storage at ambient temperature.
TABLE-US-00006 TABLE 6 Formula 4: Cream PHASE INGREDIENT % (w/v) 1
Cyclopentasiloxane 7.00 1 Cyclopentasiloxane/ 10.00 PEG/PPG-18/18
Dimethicone 1 Dimethicone 2.00 1 Polysorbate 80 0.50 1
Phenoxyethanol/Caprylyl Glycol/ 1.00 Ethylhexylglycerin/Hexylene
Glycol 2 Water 66.10 2 Glycerin 6.00 2 Disodium EDTA 0.10 2 Sodium
Metabisulfite 0.10 2 Hydroquinone 4.00 2 Ergothioneine 0.30 3
Micrococcus lysate 1.00 3 Arabidposis Extract/Lecithin/Water/ 1.00
Phenoxyethanol 3 Evodia Rutaecapra Fruit Extract/ 1.00 Butylene
Glycol/Phenoxyethanol Total 100.00
[0077] Procedure: Combine phase 1 ingredients in main kettle and
mix until uniform. In side kettle, combine phase 2 ingredients and
heat to 40.degree. C. Very slowly add phase 2 to phase 1 and mix
well. The rate of addition determines emulsion stability and
viscosity. Therefore, phase 2 is best introduced to phase 1 using a
funnel to control rate. A rate of 2.5-3 g/minute was used in this 1
kilo lab batch. Slowly add phase 3 ingredients, one at a time, to
the batch, mixing well after each addition. Switch to a Silverson
mixer, and mix for 1-3 minutes at about 2500-3500 rpm. The product
has a final pH of about 6.2 and a viscosity of about 90,000-110,000
cps. The preparation of this 0.3% ergothioneine in a water and
silicone base lotion, included manufacturing steps in which no NaOH
was used, and ergothioneine was added with sodium metabisulfite at
40.degree. C. It did not develop any fishy, amine smell after
storage at 50.degree. C. for 60 days, or over one year storage at
ambient temperatures.
Example 3
Formation of Fishy Amine Odor by Addition of Sodium Hydroxide
[0078] Four samples of a 5 ml ergothioneine solution at 0.1% (w/v)
were prepared in glass test tubes with screw caps. To the first was
added 5 ml of water, to the second 5 ml of 0.2% (w/v) sodium
metabisulfite, to the third 4 ml of water and 1 ml of 1N sodium
hydroxide, and to the fourth 5 ml of 0.2% (w/v) citric acid. The pH
of each sample was measured using litmus paper, and they were all
pH 5, except for the third sample which had a pH of approximately
10. The samples were boiled for 10 minutes and each one was
smelled. The third tube, with sodium hydroxide, had the fishy,
amine odor characteristic of ergothioneine solutions after extended
storage. None of the other tubes had this odor.
[0079] This experiment replicated the appearance of the fishy amine
odor using a simple and fast method of briefly boiling with sodium
hydroxide. None of the other components alone produced the fishy
amine odor. This allowed the testing of methods to inhibit the
formation of the odor and to suppress the odor after formation.
Example 4
Preventing Formation of Trimethylamine Odor by Addition of
Compounds Before Heating with Sodium Hydroxide
[0080] Samples were prepared in glass tubes with screw caps with
about 0.05% (w/v) ergothioneine in water, a test sample, at the
indicated concentration, and 0.1N NaOH (except for the first tube,
in which NaOH was omitted as a negative control). The pH of each
sample was determined by litmus paper, and then the tubes capped
and placed in a boiling water bath for 10 minutes. The tubes were
then cooled to room temperature, and then each one was uncapped and
smelled by three testers, each of whom scored the odor. The scores
were converted to a scale of 0 (no odor) to 3 (strong fishy
trimethylamine odor) and then averaged. A sample of each was then
assayed for ergothioneine concentration by the method of Example 1.
The results are shown in Table 7.
TABLE-US-00007 TABLE 7 Ergothioneine concentration and
trimethylamine odor rating in treated samples % EGT Test sample
Concentration pH (w/v) Odor None 0 5 0.050 0.00 NaOH without 0 11.5
0.052 3.00 trimethylamine binder trimethylamine binders added to
NaOH: Citric Acid 0.2% 8 0.054 0.00 0.05% 10 0.053 2.00 0.01% 10
0.052 2.33 Ascorbic Acid 0.2% 5 0.047 0.00 0.05% 10.5 0.050 1.67
0.01% 10.5 0.050 1.33 Boric Acid 0.2% 8.5 0.052 0.00 0.05% 10 0.051
1.33 0.01% 10.5 0.050 1.33 Acetic Acid 0.2% 4 0.052 0.00 0.05% 8
0.052 0.67 0.01% 10.5 0.050 1.67 Phosphoric 0.2% 2.5 0.058 0.00
Acid 0.05% 5.5 0.054 0.33 0.01% 10.5 0.052 1.33 Hydrochloric 0.01%
10 0.050 0.67 acid 0.0001% 11.5 0.050 0.67 Sodium 0.2% 6.5 0.029
0.00 Metabisulfite 0.05% 9.5 0.037 1.00 0.01% 10 0.044 1.67
[0081] The results show that the appearance of the fishy, amine
odor is not related to a measurable change in the concentration of
ergothioneine. The formation of the odor can be prevented or
reduced by the use of acids, particularly weak acids, and sodium
metabisulfite.
[0082] The appearance of the odor is generally a function of pH, so
that protection against the odor generally occurs at lower pH. At
equivalent pH, some acids are more efficacious at protecting
against odor formation; for example, boric.gtoreq.acetic>citric
acid at pH 8.0-8.5 At equal concentrations, some acids mixed with
0.1N NaOH produce a solution with a lower pH than others; for
example, phosphoric<acetic<ascorbic<citric<boric. This
is roughly the order of acid strength. The weaker acids appeared to
be able to prevent the formation of the odor at higher pH than
stronger acids.
Example 5
Preventing Formation of Fishy Trimethylamine Odor by Addition of
Compounds after Heating with Sodium Hydroxide
[0083] Samples were prepared in glass tubes with screw caps with 2
mM (0.046% w/v) ergothioneine in water and 0.1N NaOH (except for
the first sample, from which the NaOH was omitted as a negative
control). The tubes were capped and placed in a boiling water bath
for 10 minutes, and then cooled to room temperature. The test
samples were added to the indicated concentration, mixed and held
for a few minutes (e.g. no more than 5 minutes). Then each one was
uncapped and smelled by three testers, each of whom scored the odor
on a scale of 0 (no odor) to 3 (strong fishy, amine odor) and then
averaged. The results are shown in Table 8.
TABLE-US-00008 TABLE 8 Odor scores of mixtures containing 2 mM
(0.046%) ergothioneine Test sample Concentration Odor None 0 0.00
NaOH without 0 3.00 trimethylamine binder Trimethylamine binder
added to NaOH: Citric Acid 0.2% 1.33 0.05% 2.33 0.01% 2.33 Ascorbic
Acid 0.2% 0.00 0.05% 1.00 0.01% 2.67 Boric Acid 0.2% 1.67 0.05%
2.67 0.01% 2.67 Acetic Acid 0.2% 0.00 0.05% 1.33 0.01% 1.33
Phosphoric 0.2% 0.00 Acid 0.05% 1.00 0.01% 2.33 Hydrochloric 0.01%
1.00 acid 0.0001% 2.00 Sodium 0.2% 1.00 Metabisulfite 0.05% 2.00
0.01% 2.00
[0084] The results of this experiment show that the test samples,
especially the weak acids, may be added after the formation of the
odor to prevent or reduce its detection.
Example 6
Prevention of Fishy Amine Odor by Formation of Addition Products
with Sulfur Dioxide or Sodium Metabisulfite
[0085] Examples 4 and 5 show that sodium metabisulfite was able to
reduce or eliminate the fishy trimethylamine odor when added either
before or after the formation of trimethylamine. However, the
amount of ergothioneine detected by HPLC appeared to be
significantly reduced after addition of sodium metabisulfite.
Further study showed that the apparent reduction in the amount of
ergothioneine occurred within 10 minutes at room temperature
without boiling, suggesting this may not be a covalent
reaction.
[0086] Sodium metabisulfite generates sulfur dioxide in solution.
It is a convenient source of sulfur dioxide gas, which can be toxic
because it is readily converted to sulfuric acid. Sulfur dioxide
acts similarly to weak bases in binding to trimethylamine to form a
water-soluble and much less volatile trimethylamine addition
compound, as described by Bright and Fernelius in 1946. J. Russell
Bright and W. Conard Fernelius. "Addition compound of sulfur
dioxide and trimethylamine", Inorganic Syntheses, Volume II,
Chapter VI. Ed. W. Conard Fernelius. McGraw-Hill Book Company, Inc.
pp. 159-161, 1946.
[0087] The apparent loss of the ergothioneine immediately after
addition of sodium metabisulfite is due to the formation of another
reversible addition product between sulfur dioxide and
ergothioneine, as described by Balaban and King in 1927. Isidore
Elkanah Balaban and Harold King. "Gold and mercury derivatives of
2-thiol-glyoxalines. Mechanism of the oxidation of
2-thiolglyoxalines to glyoxalines". J. Chem. Soc. (1927) pp
1858-1874. This addition product, at sufficiently high
concentration, has a yellow color.
[0088] To demonstrate this chemical reaction, sample vials were
prepared with a 1.25% solution of ergothioneine in water with
increasing amounts of sodium metabisulfite from 2.5% to 50%. The
vials were sealed and mixing the samples was avoided. The samples
were incubated and color development observed. The solutions were
examined at 1 hour, and the optical density at 405 nm was measured
at 2 hours (Table 9). As the concentration of sodium metabisulfite
increased, the intensity of the yellow color, and the OD.sub.405
also increased.
TABLE-US-00009 TABLE 9 Formation of yellow color in a solution of
ergothioneine and sodium metabisulfite Sodium Ergothioneine
metabisulfite Color description at 1 h OD.sub.405 at 2 hr 2.5% 0
Colorless 0.004 1.25% 2.5% Colorless 0.044 1.25% 5.0% Near
colorless 0.104 1.25% 7.5% Yellow tinge 0.186 1.25% 10% Slight
yellow 0.265 1.25% 12.5% Light yellow 0.456 1.25% 15% Yellow 0.593
1.25% 17.5% Yellow 0.966 1.25% 20% Light lemon yellow 1.993 1.25%
22.5% Intense lemon yellow 2.873 1.25% 25% Intense lemon yellow
3.126
[0089] It was further observed that the addition product, and hence
the yellow color, is entirely reversible by driving off the sulfur
dioxide gas. To demonstrate this chemical reaction, a solution of
1.25% ergothioneine and 5% sodium metabisulfite turned intense
lemon yellow after one week at room temperature in a capped tube.
The cap was removed and the solution was then shaken for 3 to 4
minutes and ambient air was bubbled through it to drive out the
dissolved sulfur dioxide gas. After 6 to 7 minutes the solution
turned nearly colorless with a yellow tinge. Analysis of the
ergothioneine content of the solutions by HPLC shows a reduction in
concentration with the formation of the yellow color, and a partial
restoration of the ergothioneine concentration with the loss of the
yellow color. Therefore the loss of ergothioneine after addition of
sodium metabisulfite is due to formation of an addition product
which is reversible, and the ergothioneine is not permanently
lost.
Example 7
Preventing Formation of Fishy Amine Odor
TABLE-US-00010 [0090] TABLE 10 Formula 5: Concentrate PHASE
INGREDIENT % (w/v) 1 Cyclopentasiloxane 10.00 1
Cyclopentasiloxane/PEG/PPG-18/18 10.00 Dimethicone 1 Dimethicone
4.00 1 Polysorbate 80 0.50 1 Phenoxyethanol/Caprylyl Glycol/ 1.00
Ethylhexylglycerine/Hexylene Glycol 2 Water 62.30 2 Glycerin 6.00 2
Tetrasodium EDTA 0.10 2 Undecylenoyl Phenylalanine 2.00 2 Sodium
Hydroxide 1.00 2A Ergothioneine 0.10 2A Citric Acid 0.20 3 Sodium
Hydroxide * 3 Citric Acid * 4 Micrococcus lysate 1.20 4 Arabidposis
Extract/Lecithin/Water/ 1.00 Phenoxyethanol 4 Evodia Rutaecapra
Fruit Extract/ 1.00 Butylene Glycol/Phenoxyethanol Total 100.00 *
(amount sufficient to adjust pH to 6.8-7.4)
[0091] Procedure: In the main kettle, combine phase 1 ingredients
and mix until uniform. In a side kettle, combine phase 2
ingredients and heat to 65.degree. C. Cool phase 2 to 25.degree. C.
and adjust pH to 6.8-7.4 and clear using phase 3 ingredients. At
25.degree. C., add phase 2A ingredients to phase 2 with propeller
mixing. Very slowly add phase 2 to phase 1 and mix well. The rate
of addition determines emulsion stability and viscosity. It is best
added with a funnel to control rate. (A rate of 1-2 g/minute was
used in the 1 kilo lab batch.) Slowly add phase 4 ingredients and
mix until uniform. Switch to a Silverson mixer, and mix for 1-3
minutes at approximately 2500-3500 rpm. The product has a final pH
of about 6.8-7.2, a viscosity of 15,000-20,000 cps and a specific
gravity of 1.01.+-.0.02. The preparation of this 0.1% ergothioneine
in water and magnesium aluminum silicate base with 0.2% citric acid
included a manufacturing step in which the ergothioneine was added
in a phase at the end when the formula had cooled to 25.degree. C.
It had no odor immediately after manufacturing and did not develop
an odor after heating for 60 days at 40.degree. C. or after six
months of storage at ambient temperature.
[0092] As used herein, "ameliorate" means, with respect to the
trimethylamine, to improve the trimethylamine odor by reducing,
inhibiting, preventing, or eliminated it; "treatment composition"
means a product for use in treatment mammals, and may include
ingestible, topical or injectible products; and "pre-blend" means a
composition that may be in the form of a solution, suspension,
emulsion or anhydrous composition, that is formed separately by
combining the ingredients present in the pre-blend, and then
incorporating the pre-blend into the treatment composition during
the treatment composition formulation process. By use of the term
"consisting essentially of", it is intended that the composition or
method does not include any component or step which would
materially affect the basic and novel characteristics of the
invention.
* * * * *