Note: Descriptions are shown in the official language in which they were submitted.
CA 02577439 2007-02-20
Creatine-Fatty Acids
Field of the Invention
The present invention relates to structures and synthesis of creatine-fatty
acid compounds bound via an anhydride linkage. Another aspect of the present
invention relates to a compound comprising a creatine molecule bound to a
fatty
acid, wherein the fatty acid is preferably a saturated fatty acid and bound to
the
creatine via an anhydride linkage.
Background of the Invention
Creatine is a naturally occurring amino acid derived from the amino acids
glycine, arginine, and methionine. Although it is found in meat and fish, it
is also
synthesized by humans. Creatine is predominantly used as a fuel source in
muscle. About 65% of creatine is stored in the musculature of mammals as
phosphocreatine (creatine bound to a phosphate molecule).
Muscular contractions are fueled by the dephosphorylation of adenosine
triphosphate (ATP) to produce adenosine diphosphate (ADP). In the absence of
a mechanism to replenish ATP stores, the supply of ATP would be totally
consumed in 1-2 seconds. Phosphocreatine serves as a major source of
phosphate from which ADP is regenerated to ATP. Within six seconds following
the commencement of exercise, muscular concentrations of phosphocreatine
drop by almost 50%. Creatine supplementation has been shown to increase the
concentration of creatine in the muscle (Harris RC, Soderfund K, Hultman E.
Elevation of creatine in resting and exercised muscle of normal subjects by
creatine supplementation. Clin Sci (Lond). 1992 Sep;83(3):367-74) and further,
1
CA 02577439 2007-02-20
the supplementation enables an increase in the resynthesis of phosphocreatine
(Greenhaff PL, Bodin K, Soderlund K, Hultman E. Effect of oral creatine
supplementation on skeletal muscle phosphocreatine resynthesis. Am J Physiol.
i 994 May.266(5 Pt I j-E725-30) leading to a rapid replenishment of ATP within
the first two minutes following the commencement of exercise. Through this
mechanism, creatine is able to improve strength and reduce fatigue (Greenhaff
PL, Casey A, Short AH, Harris R, Soderlund K, Hultman E. Influence of oral
creatine supplementation of muscle torque during repeated bouts of maximal
voluntary exercise in man. Clin Sci (Lond). 1993 May;84(5):565-71).
The beneficial effects of creatine supplementation with regard to skeletal
muscle are apparently not restricted to the role of creatine in energy
metabolism.
It has been shown that creatine supplementation in combination with strength
training results in specific, measurable physiological changes in skeletal
muscle
compared to strength training alone. For example, creatine supplementation
amplifies the strength training-induced increase of human skeletal satellite
cells
as well as the number of myonuclei in human skeletal muscle fibres (Olsen S,
Aagaard P, Kadi F, Tufekovic G, Verney J, Olesen JL. Suetta C, Kjaer M.
Creatine supplementation augments the increase in satellite cell and myonuclei
number in human skeletal muscle induced by strength training. J Physiol. 2006
Jun 1;573(Pt 2):525-34). Satellite cells are the stem cells of adult muscle.
They
are normally maintained in a quiescent state and become activated to fulfill
roles
of routine maintenance, repair and hypertrophy (Zammit PS, Partridge TA,
Yablonka-Reuveni Z. The Skeletal Muscle Satellite Cell: The Stem Cell That
2
CA 02577439 2007-02-20
Carne, In r=rorn the Cold. J Histocherrm Cytochem. 2006 Aug 9). 'True' muscle
hypertrophy can be defined as "as an increase in fiber diameter without an
apparent increase in the number of muscle fibers, accompanied by enhanced
protein synthesis and augmented contractile force" (Sartorelli V, Fulco M.
Molccuior and cellular determinants of skeietal muscle atrophy and
hypertrophy.
Sci STKE. 2004 Jul 27,2004(244):re11). Postnatal muscle growth involves both
myofiber hypertrophy and increased numbers of myonuclei - the source of which
are satellite cells (Olsen S, Aagaard P, Kadi F, Tufekovic G, Verney J, Olesen
JL., Suetta C, Kjaer lVl. Creatine supplementation augments the increase in
saieiiite cell dnd niyonucIei riumber in human skeletal rriuscle induced by
strength training. J Physioi. 2006 Jun 1;573(Pt 2):525-34).
Although creatine is used predominantly in muscle cells and most of the
total creatine pool is found in muscle, creatine is actually synthesized in
the liver
and pancreas. Thus, the musculature's creatine concentration is maintained by
the uptake of creatine from the blood stream regardless of whether the source
of
creatine is endogenous, i.e. synthesized by the liver or pancreas, or dietary,
i.e.
natural food sources or supplemental sources. The creatine content of an
average 70 kg male is approximately 120 g with about 2 g being excreted as
creatinine per day (Williams MH, Branch JD. Creatine supplementation and
exercise performance: an update. J Am Call Nutr. 1998 Jun;17(3):216-34). A
typical omnivorous diet supplies approximately 1 g of creatine daily, while
diets
higher in meat and fish will supply more creatine. As a point of reference, a
500
g uncooked steak contains about 2 g of creatine which equates to more than two
3
CA 02577439 2007-02-20
8 oz. steaks per day. Since most studies examining creatine supplementation
employ dosages ranging from 2-20 g per day it is unrealistic to significantly
increase muscle creatine stores through merely food sources alone. Therefore,
supplemental sources of creatine are an integral component of increasing, and
subsequently maintaining supraphysiological, muscular creatine levels.
Creatine supplementation, thus results in positive physiological effects on
skeletal muscle, such as: performance improvements during brief high-intensity
anaerobic exercise, increased strength and enhanced muscle growth.
Creatine monohydrate is a commonly used supplement. Creatine
monohydrate is soluble in water at a rate of 75 ml of water per gram of
creatine.
Ingestion of creatine monohydrate, therefore, requires large amounts of water
to
be co-ingested. Additionally, in aqueous solutions creatine is known to
convert to
creatinine via an irreversible, pH-dependent, non-enzymatic reaction. Aqueous
and alkaline solutions contain an equilibrium mixture of creatine and
creatinine.
In acidic solutions, on the other hand, the formation of creatinine is
complete.
Creatinine is devoid of the ergogenic beneficial effects of creatine. It is
therefore
desirable to provide, for use in individuals, e.g. animals and humans, forms
and
derivatives of creatine with improved characteristics such as stability and
solubility. Furthermore, it would be advantageous to do so in a manner that
provides additional functionality as compared to creatine monohydrate alone.
The manufacture of hydrosoluble creatine salts with various organic acids
have been described. U.S. Pat. No. 5,886,040, purports to describe a creatine
pyruvate salt with enhanced palatability which is resistant to acid
hydrolysis.
4
CA 02577439 2007-09-05
U.S. Patent No. 5,973,199, purports to describe hydrosoluble organic salts of
creatine as single combination of one mole of creatine monohydrate with one
mole of
the following organic acids: citrate, malate, fumarate and tartarate
individually. The
resultant salts described therein are claimed to be from 3 to 15 times more
soluble,
in aqueous solution, than creatine itself.
U.S. Pat. No. 6,166,249, purports to describe a creatine pyruvic acid salt
that
is highly stable and soluble. It is further purported that the pyruvate
included in the
salt may be useful to treat obesity, prevent the formation of free radicals
and
enhance long-term performance.
U.S. Pat. No. 6,211,407 purports to describe dicreatine and tricreatine
citrates
and a method of making the same. These dicreatine and tricreatine salts are
claimed
to be stable in acidic solutions, thus hampering the undesirable conversion of
creatine to creatinine.
U.S. Pat. No. 6,838,562, purports to describe a process for the synthesis of
mono, di, or tricreatine orotic acid, thioorotic acid, and dihydroorotic acid
salts which
are claimed to have increased oral absorption and bioavailability due to an
inherent
stability in aqueous solution. It is further claimed that the heterocyclic
acid portion of
the salt acts synergistically with creatine.
U.S. Pat. No. 7,109,373 purports to describe creatine salts of dicarboxylic
acids with enhanced aqueous solubility. The above disclosed patents recite
creatine
salts, methods of synthesis of the salts, and uses thereof. However, nothing
in any
of the disclosed patents
CA 02577439 2007-02-20
teaches, suggests or discloses a compound comprising a creatine molecule
bound to a fatty acid.
In addition to salts, creatine esters have also been described. U.S. Pat.
No. 6,897,334 describes method for producing creatine esters with lower
alcohols i.e. one to four carbon atoms, using acid catalysts. It is stated
that
creatine esters are more soluble than creatine. It is further stated that the
protection of the carboxylic acid moiety of the creatine molecule by ester-
formation stabilizes the compound by preventing its conversion to creatinine.
The creatine esters are said to be converted into creatine by esterases i.e.
enzymes that cleave ester bonds, found in a variety of cells and biological
fluids.
Fatty acids are carboxylic acids, often containing a long, unbranched
chain of carbon atoms and are either saturated or unsaturated. Saturated fatty
acids do not contain double bonds or other functional groups, but contain the
maximum num.ber of hydrogen atoms, with the exception of the carboxylic acid
group. In contrast, unsaturated fatty acids contain one or more double bonds
between adjacent carbon atoms, of the chains, in cis or trans configuration
The human body can produce all but two of the fatty acids it requires,
thus, essential fatty acids are fatty acids that must be obtained from food
sources
due to an inability of the body to synthesize them, yet are required for
normal
biological function. The essential fatty acids being linoleic acid and a-
linolenic
acid.
Examples of saturated fatty acids include, but are not limited to myristic or
tetradecanoic acid, paimitic or hexadecanoic acid, stearic or octadecanoic
acid,
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CA 02577439 2007-02-20
arachidic or eicosanoic acid, behenic or docosanoic acid, butyric or butanoic
acid, caproic or hexanoic acid, caprylic or octanoic acid, capric or decanoic
acid,
and lauric or dodecanoic acid, wherein the aforementioned comprise from at
least 4 carbons to 22 carbons in the chain.
Examples of unsaturated fatty acids include, but are not limited to oleic
acid, linoleic acid, linolenic acid, arachidonic acid, palmitoleic acid,
eicosapentaenoic acid, docosahexaenoic acid and erucic acid, wherein the
aforementioned comprise from at least 4 carbons to 22 carbons in the chain.
Fatty acids are capable of undergoing chemical reactions common to
carboxylic acids. Of particular relevance to the present invention are the
formation of salts and the formation of esters. The majority of the above
referenced patents are creatine salts. These salts, esterification via
carboxylate
reactivity, may essentially be formed, as disclosed in U.S. Pat. No.
7,109,373,
through a relatively simple reaction by mixing a molar excess of creatine or
derivative thereof with an aqueous dicarboxylic acid and heating from room
temperature to about 50 C.
Alternatively, a creatine-fatty acid may be synthesized through ester
formation. The formation of creatine esters has been described (Dox AW, Yoder
L, Esterification of Creatine. J, Biol. Chem. 1922, 67, 671-673). These are
typically formed by reacting creatine with an alcohol in the presence of an
acid
catalyst at temperatures from 35 C to 50 C as disclosed in U.S. Pat. No.
6,897,334.
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CA 02577439 2007-02-20
While the above referenced creatine compounds have attempted to
address issues such as stability and solubility in addition to, and in some
cases,
to add increased functionality as compared to creatine alone, no description
has
yet been made of any creatine-fatty acid compound, particularly that
comprising
a saturated fatty acid.
Summary of the Invention
In the present invention, compounds are disclosed, where the compounds
comprise a molecule of creatine bound to a fatty acid, via an anhydride
linkage,
and having a structure of Formula 1:
Formula 1
O O
li II
HzC' C~O~C" R
CH3
NH2
where:
R is an alkyl group, preferably saturated, and containing from about 3 to a
maximum of 21 carbons.
Another aspect of the invention comprises the use of a saturated fatty acid
in the production of compounds disclosed herein.
A further aspect of the present invention comprises the use of an
unsaturated fatty in the production of compounds disclosed herein.
8
CA 02577439 2007-02-20
Detailed Description of the Invention
In the following description, for the purposes of explanation, numerous
specific details are set forth in order to provide a thorough understanding of
the
present invention. It will be apparent, however, to one skilled in the art
that the
present invention may be practiced without these specific details.
The present invention relates to structures and synthesis of creatine-fatty
acid compounds bound via an anhydride linkage.. In addition, specific benefits
are conferred by the particular fatty acid used to form the compounds in
addition
to, and separate from, the creatine substituent.
As used herein, the term 'fatty acid' includes both saturated, i.e. an alkane
chain as known in the art, having no double bonds between carbons of the chain
and having the maximum number of hydrogen atoms, and unsaturated, i.e. an
alkene or alkyne chain, having at least one double or alternatively triple
bond
between carbons of the chain, respectively, and further terminating the chain
in a
carboxylic acid as is commonly known in the art, wherein the hydrocarbon chain
is not less then four carbon atoms. Furthermore, essential fatty acids are
herein
understood to be included by the term 'fatty acid'.
As used herein, "creatine" refers to the chemical N-methyl-N-guanyl
Glycine, (CAS Registry No. 57-00-1), also known as, (alpha-methyl guanido)
acetic acid, N-(aminoiminomethyl)-N-glycine, Methylglycocyamine,
Methylguanidoacetic Acid, or N-Methyl-N-guanylglycine. Additionally, as used
herein, "creatine" also includes derivatives of creatine such as esters, and
9
CA 02577439 2007-02-20
amides, and salts, as well as other derivatives, including derivatives having
pharmacoproperties upon metabolism to an active form.
According to the present invention, the compounds disclosed herein
comprise a creatine molecule bound to a fatty acid, wherein the fatty acid is
preferably a saturated fatty acid. Furthermore, the creatine and fatty acid
being
bound by an anhydride linkage and having a structure according to Formula 1.
The aforementioned compound being prepared according to the reaction as set
forth for the purposes of the description in Scheme 1:
Scheme 1
0 + 0 Step 1 0
35 C - 50 C I (
ROH XX 0.5-2h R" C~X
2 3 4 0 0
II II
MO~C" CH Step 2 HO~C~CH
Step 3 I 2 1)NaOH, HZO I Z
where: H3C' NC -,NH 2)evap. H20 H3C' N~C-,NH
R = alkane or alkene (C = 3 to 21) NH2 NH2
X=CI,Br,F,orI 6 5
M= Na, K, Li, or NH4 0 0
II II
H2C' C, OR
HN ~ ~N~
~C CH3
NH2
1
With reference to Scheme 1, in Step 1 an acyl halide (4) is produced via
reaction of a fatty acid (2) with a thionyl halide (3).
In various embodiments of the present invention, the fatty acid of (2) is
selected from the saturated fatty acid group comprising butyric or butanoic
acid,
caproic or hexanoic acid, caprylic or octanoic acid, capric or decanoic acid,
lauric
CA 02577439 2007-02-20
or dodecanoic acid, myristic or tetradecanoic acid, paimitic or hexadecanoic
acid,
stearic or octadecanoic acid, arachidic or eicosanoic acid, and behenic or
docosanoic acid.
In alternative embodiments, of the present invention, the fatty acid of (2) is
selected from the unsaturated fatty acid group comprising oleic acid, linoleic
acid,
linolenic acid, arachidonic acid, paimitoleic acid, eicosapentaenoic acid,
docosahexaenoic acid, and erucic acid.
Furthermore, the thionyl halide of (3) is selected from the group consisting
of fluorine, chlorine, bromine, and iodine, the preferred method using
chlorine or
bromine.
The above reaction proceeds under conditions of heat ranging between
from about 35 C to about 50 C and stirring over a period from about 0.5 hours
to
about 2 hours during which time the gases sulfur dioxide and acidic gas,
wherein
the acidic gas species is dependent on the species of thionyl halide employed,
are evolved. Preferably, the reaction proceeds at 45 C for 1.5 hours.
Step 2 of Scheme 1 entails the neutralization of the carboxylic acid of the
creatine portion through the addition of an inorganic base. The inorganic base
is
selected from the group comprising sodium hydroxide, potassium hydroxide,
lithium hydroxide, ammonium hydroxide, sodium carbonate. Preferred inorganic
bases for the purposes of the present invention are sodium hydroxide and
potassium hydroxide.
Neutralization, as described above, is followed by the evaporation of
water, resulting in the isolation of the corresponding salt. For example,
11
CA 02577439 2007-02-20
potassium hydroxide, when used as the inorganic base, results in the
production
of the potassium creatine salt.
Step 3 of Scheme I involves the drop wise addition of the prepared acyl
halide (4) to the creatine salt (6) in a cooled flask and subsequent
purification by
two rounds of distillation to yield the desired anhydride compound (1), the
anhydride compound being a creatine fatty acid compound of the present
invention.
In various embodiments, according to aforementioned, using the saturated
fatty acids, the following compounds are produced produced: butyric 2-(1-
methylguanidino)acetic anhydride, hexanoic 2-(1-methyiguanidino)acetic
anhydride, 2-(1-methylguanidino)acetic octanoic anhydride, decanoic 2-(1-
methylguanidino)acetic anhydride, 2-(1-methyiguanidino)acetic tetradecanoic
anhydride, 2-(1-methylguanidino)acetic paimitic anhydride, icosanoic 2-(1-
methylguanidino)acetic anhydride, and docosanoic 2-(1-methylguanidino)acetic
anhydride.
In additional embodiments, according to aforementioned, using the
unsaturated fatty acids, the following compounds are produced produced: (Z)-
hexadec-9-enoic 2-(1-methylguanidino)acetic anhydride, 2-(1-
methylguanidino)acetic oleic anhydride, (Z)-docos-13-enoic 2(1-
methylguanidino)acetic anhydride, 2-(1 -methylguanidino)acetic (9Z, 1 2Z)-
octadeca-9,12-dienoic anhydride, 2-(1-methylguanidino)acetic (9Z,12Z,15Z)-
octadeca-9,12,15-trienoic anhydride, 2-(1-methylguanidino)acetic (6Z,9Z,12Z)-
octadeca-6,9,12-trienoic anhydride, (5Z,8Z,11 Z,14Z)-icosa-5,8,11,14-
tetraenoic
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CA 02577439 2007-02-20
2(1-methylguanidino)acetic anhydride, (5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-
pentaenoic 2(1-methylguanidino)acetic anhydride, 2(1-methylguanidino)acetic
(8Z,11 Z, 1 4Z, 1 7Z,20Z)-tricosa-5,8,11,14,17,20-hexaenoic anhydride.
The following examples illustrate specific creatine-fatty acids and routes of
synthesis thereof. One of skill in the art may envision various other
combinations
within the scope of the present invention, considering examples with reference
to
the specification herein provided.
Example 1
Butyric 2-(1-methylguanidino)acetic anhydride
0 0
II li H2
H C~C~O~C~C" C~CH
2I H2 3
HN
CH3
NH2
In a dry 2-necked, round bottomed flask, equipped with a magnetic stirrer
and fixed with a separatory funnel, containing 8.75mi (120mmol) of thionyl
chloride, and a water condenser, is placed 9.05ml (100mmol) of butanoic acid.
Addition of the thionyl chloride is completed with heating to about 40 C over
the
course of about 30 minutes. When addition of the thionyl chloride is complete
the mixture is heated and stirred for an additional 30 minutes. The water
condenser is then replaced with a distillation side arm condenser and the
crude
mixture is distilled. The crude distillate in the receiving flask is then
fractionally
distilled to obtain the acyl chloride, butyryl chloride.
Separately, in a single-necked, round bottomed flask, equipped with a
magnetic stirrer, 6.56g (50mmol) of creatine is dissolved in 500m1 of water.
To
13
CA 02577439 2007-02-20
this is added 55m1 of 1 M sodium hydroxide with vigorous stirring, until heat
production ceases. At this point the water is removed by evaporation to yield
the
carboxylate salt, sodium 2-(1-methylguanidino)acetate, shown below.
O
11
NaO' C" i CH2
H C' N~C~NH
3 I
NH2
Finally, in a dry 2-necked, round bottomed flask, fixed with a separatory
funnel, containing 6.39g (60mmol) of the prepared butyryl chloride, and side
arm
water condenser fixed with a dry receiving flask, is placed 12.08g (66mmol) of
sodium 2-(1-methylguanidino)acetate. The round bottomed flask is placed in an
ice bath and the butyryl chloride is added drop wise. After addition is
completed
the mixture is shaken and the ice bath is replaced by a heating mantle. The
flask
is then heated until no more solution is dropping into the receiving flask.
This
crude distillate is then further fractionally distilled to yield butyric 2-(1-
methylguanidino)acetic anhydride.
Example 2
Hexanoic 2-(1-methylguanidino)acetic anhydride
O 0
H2
HZC' C" O~C~C CH3
I HZ 2
HN~ ~N~
~C CH3
NH2
In a dry 2-necked, round bottomed flask, equipped with a magnetic stirrer
and fixed with a separatory funnel, containing 6.97m1 (90mmol) of thionyl
14
CA 02577439 2007-02-20
bromide, and a water condenser, is placed 5.68ml (45mmol) of hexanoic acid.
Addition of the thionyl bromide is completed with heating to about 50 C over
the
course of about 50 minutes. When addition of the thionyl bromide is complete
the mixture is heated and stirred for an additional hour. The water condenser
is
then replaced with a distillation side arm condenser and the crude mixture is
distilled. The crude distillate in the receiving flask is then fractionally
distilled to
obtain the acyl bromide, hexanoyl bromide.
Separately, in a single-necked, round bottomed flask, equipped with a
magnetic stirrer, 6.56g (50mmol) of creatine is dissolved in 500m1 of water.
To
this is added 55m1 of 1 M sodium hydroxide with vigorous stirring, until heat
production ceases. At this point the water is removed by evaporation to yield
the
carboxylate salt, sodium 2-(1-methylguanidino)acetate, shown below.
O
11
NaO"C"CH2
H C ' N~C ~NH
3 i
NH2
Finally, in a dry 2-necked, round bottomed flask, fixed with a separatory
funnel, containing 10.81g (60mmol) of the prepared hexanoyl bromide, and side
arm water condenser fixed with a dry receiving flask, is placed 13.18g
(72mmol)
of sodium 2-(1-methylguanidino)acetate. The round bottomed flask is placed in
an ice bath and the hexanoyl bromide is added drop wise. After addition is
completed the mixture is shaken and the ice bath is replaced by a heating
mantle. The flask is then heated until no more solution is dropping into the
CA 02577439 2007-02-20
receiving flask. This crude distillate is then further fractionally distilled
to yield
hexanoic 2-(1-methylguanidino)acetic anhydride.
Example 3
Dodecanoic 2-(1-methylguanidino)acetic anhydride
0 0 H2
HzC' C" 0"C~C~C CH3
I H2 5
HN C'_N'CH3
I
NH2
In a dry 2-necked, round bottomed flask, equipped with a magnetic stirrer
and fixed with a separatory funnel, containing 5.85m1 (80mmol) of thionyl
chloride, and a water condenser, is placed 10.02g (50mmol) of dodecanoic acid.
Addition of the thionyl chloride is completed with heating to about 45 C over
the
course of about 40 minutes. When addition of the thionyl chloride is complete
the mixture is heated and stirred for an additional 50 minutes. The water
condenser is then replaced with a distillation side arm condenser and the
crude
mixture is distilled. The crude distillate in the receiving flask is then
fractionally
distilled to obtain the acyl chloride, dodecanoyl chloride.
Separately, in a single-necked, round bottomed flask, equipped with a
magnetic stirrer, 7.87g (60mmol) of creatine is dissolved in 600m1 of water.
To
this is added 78ml of 1 M ammonium hydroxide with vigorous stirring, until
heat
production ceases. At this point the water is removed by evaporation to yield
the
carboxylate salt, ammonium 2-(1-methylguanidino)acetate, shown below.
16
CA 02577439 2007-02-20
0
11
HdNO' C" CH2
H C' N,, C~NH
3
NH2
Finally, in a dry 2-necked, round bottomed flask, fixed with a separatory
funnel, containing 15.31g (70mmol) of the prepared dodecanoyl chloride, and
side arm water condenser fixed with a dry receiving flask, is placed 12.44g
(84mmol) of ammonium 2-(1-methylguanidino)acetate. The round bottomed
flask is piaced in an ice bath and the dodecanoyl chloride is added drop wise.
After addition is completed the mixture is shaken and the ice bath is replaced
by
a heating mantle. The flask is then heated until no more solution is dropping
into
the receiving flask. This crude distillate is then further fractionally
distilled to
yield dodecanoic 2-(1-methylguanidino)acetic anhydride.
Example 4
2-(1-methylguanidino)acetic stearic anhydride
0 0
H2
H2C' C" O"C(C"CCH3
I H2 g
HN ~ ~N'~C CH3
NH2
In a dry 2-necked, round bottomed flask, equipped with a magnetic stirrer
and fixed with a separatory funnel, containing 4.81 ml (66mmol) of thionyl
chloride, and a water condenser, is placed 15.65g (55mmol) of stearic acid.
Addition of the thionyl chloride is completed with heating to about 45 C over
the
course of about 40 minutes. When addition of the thionyl chloride is complete
17
CA 02577439 2007-02-20
the mixture is heated and stirred for an additional 45 minutes. The water
condenser is then replaced with a distillation side arm condenser and the
crude
mixture is distilled. The crude distillate in the receiving flask is then
fractionally
distilled to obtain the acyl chloride, stearoyl chloride.
Separately, in a single-necked, round bottomed flask, equipped with a
magnetic stirrer, 7.87g (60mmol) of creatine is dissolved in 600mi of water.
To
this is added 72m1 of 1 M potassium hydroxide with vigorous stirring, until
heat
production ceases. At this point the water is removed by evaporation to yield
the
carboxylate salt, potassium 2-(1-methylguanidino)acetate, shown below.
O
I I
KOCH2
H C' N~C ~NH
3 I
NH2
Finally, in a dry 2-necked, round bottomed flask, fixed with a separatory
funnel, containing 21.27g (70mmol) of the prepared stearoyl chloride, and side
arm water condenser fixed with a dry receiving flask, is placed 23.40g
(77mmol)
of potassium 2-(1-methylguanidino)acetate. The round bottomed flask is placed
in an ice bath and the stearoyl chloride is added drop wise. After addition is
completed the mixture is shaken and the ice bath is replaced by a heating
mantle. The flask is then heated until no more solution is dropping into the
receiving flask. This crude distillate is then further fractionally distilled
to yield 2-
(1 -methylguanidino)acetic stearic anhydride.
Example 5
2-(1-methylguanidino)acetic (9Z,12Z)-octadeca-9,12-dienoic anhydride
18
CA 02577439 2007-02-20
0 0
H2 3 H H H H H2 H2
H2C~C" O"C~C"C~C,C=C,, C,C=C,, C~-C~C~C~CH3
i H2 H2 H2 H2 H2
HN~C~N~CH
3
NH2
In a dry 2-necked, round bottomed flask, equipped with a magnetic stirrer
and fixed with a separatory funnel, containing 9.35m1 (128mmol) of thionyl
chloride, and a water condenser, is placed 24.90m1 (80mmol) of linoleic acid.
Addition of the thionyl chloride is completed with heating to about 40 C over
the
course of about 40 minutes. When addition of the thionyl chloride is complete
the mixture is heated and stirred for an additional 50 minutes. The water
condenser is then replaced with a distillation side arm condenser and the
crude
mixture is distilled. The crude distillate in the receiving flask is then
fractionally
distilled to,obtain the acyl chloride, (9Z,12Z)-octadeca-9,12-dienoyl
chloride.
Separately, in a single-necked, round bottomed flask, equipped with a
magnetic stirrer, 7.87g (60mmol) of creatine is dissolved in 600m1 of water.
To
this is added 78ml of 1M ammonium hydroxide with vigorous stirring, until heat
production ceases. At this point the water is removed by evaporation to yield
the
carboxylate salt, ammonium 2-(1-methylguanidino)acetate, shown below.
O
I I
H4NO' C" CH2
H C~N~C~NH
3 I
NH2
Finally, in a dry 2-necked, round bottomed flask, fixed with a separatory
funnel, containing 17.93g (60mmol) of the prepared (9Z,12Z)-octadeca-9,12-
19
CA 02577439 2007-02-20
dienoyl chloride, and side arm water condenser fixed with a dry receiving
flask, is
placed 10.66g (72mmol) of ammonium 2-(1-methylguanidino)acetate. The round
bottomed flask is placed in an ice bath and the (9Z,12Z)-octadeca-9,12-dienoyl
chloride is added drop wise. After addition is completed the mixture is shaken
and the ice bath is replaced by a heating mantle. The flask is then heated
until
no more solution is dropping into the receiving flask. This crude distillate
is then
further fractionally distilled to yield 2-(1-methylguanidino)acetic(9Z,12Z)-
octadeca-9,12-dienoic anhydride.
Thus while not wishing to be bound by theory, it is understood that
reacting a creatine or derivative thereof with a fatty acid or derivative
thereof to
form an anhydride can be used enhance the bioavailability of the creatine or
derivative thereof by improving stabiiity of the creatine moiety in terms of
resistance to hydrolysis in the stomach and blood and by increasing solubility
and absorption. Furthermore, it is understood that, dependent upon the
specific
fatty acid, for example, saturated fatty acids form straight chains allowing
mammals to store chemical energy densely, or derivative thereof employed in
the
foregoing synthesis, additional fatty acid-specific benefits, separate from
the
creatine substituent, will be conferred.
Extensions and Alternatives
In the foregoing specification, the invention has been described with a
specific embodiment thereof; however, it will be evident that various
modifications and changes may be made thereto without departing from the
broader spirit and scope of the invention.
