U.S. patent application number 10/700424 was filed with the patent office on 2004-08-05 for synthesis of coenzyme q10, ubiquinone.
Invention is credited to West, Daniel David.
Application Number | 20040151711 10/700424 |
Document ID | / |
Family ID | 25274154 |
Filed Date | 2004-08-05 |
United States Patent
Application |
20040151711 |
Kind Code |
A1 |
West, Daniel David |
August 5, 2004 |
Synthesis of coenzyme Q10, ubiquinone
Abstract
Processes for the stereospecific synthesis of coenzyme Q10,
ubiquinone, are disclosed; a semi synthetic procedure using
solanesol derived from tobacco waste as the starting material. The
process of the invention results in high yields of isometrically
useful compositions containing the optically pure isomers.
Inventors: |
West, Daniel David;
(Rockport, MA) |
Correspondence
Address: |
Evelya M. Sommer
30th Floor
825 Third Avenue
New York
NY
10022
US
|
Family ID: |
25274154 |
Appl. No.: |
10/700424 |
Filed: |
November 5, 2003 |
Related U.S. Patent Documents
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
|
|
10700424 |
Nov 5, 2003 |
|
|
|
09837320 |
Apr 19, 2001 |
|
|
|
6686485 |
|
|
|
|
Current U.S.
Class: |
424/94.1 ;
552/307 |
Current CPC
Class: |
C07C 46/00 20130101;
C07C 45/676 20130101; C07C 46/02 20130101; C07B 2200/09 20130101;
C07C 46/02 20130101; C07C 29/40 20130101; C07C 45/676 20130101;
C07C 46/00 20130101; C07C 29/40 20130101; C07C 50/28 20130101; C07C
50/28 20130101; C07C 33/02 20130101; C07C 49/21 20130101 |
Class at
Publication: |
424/094.1 ;
552/307 |
International
Class: |
A61K 038/43; C07C
050/28 |
Claims
I claim:
1. A stereospecific synthesis of optically pure trans (E) isomer of
coenzyme Q 10 having the formula 7which comprises extracting
solanesol from tobacco dust and using said solanesol as the
starting material for carrying out the following sequence of
reactions 8separately carrying out the following reactions:
9thereafter reacting the isodecaprenol and
2,3,-dimethoxy-5-methyl-hydroquinone to form the optical pure
ubiquinone 10
2. Method of treating impaired or damaged tissue in humans and
animals which comprises administering a composition comprising as
the principal active ingredient a therapeutically effective amount
of optically pure trans (E) isomer of coenzyme Q 10
(2,3-dimethoxy-5-methyl-6-decaprenyl-be- uoquinone) in admixture
with a pharmaceutically acceptable carrier.
3. The method of claim 2 wherein said composition is administered
orally.
4. The method of claim 3 wherein said composition is administered
in an amount of 15-400 mg pro die.
5. The method of claim 3 wherein said composition is administered
in an amount of 100-200 mg pro die.
6. The method of claim 3 wherein said composition is administered
in an amount of 15-30 mg pro die.
7. The method of claim 3 wherein said composition is in tablet
form.
8. The method of claim 3 wherein said composition is in liquid
form.
9. The method of claim 2 wherein said composition is administered
by topical application.
10. The method of claim 9 wherein said composition contains the
optically pure coenyme Q 10 in an amount of 0.1-10%.
11. The method of claim 9 wherein said composition contains the
optically pure coenzyme Q 10 in an amount of 0.25-1%.
12. The method of claim 9 wherein said composition is to be used as
a cosmetic and said optically pure coenzyme Q 10 is present in an
amount of 0.0001 to 0.1%.
13. The method of claim 2 wherein said pharmaceutically acceptable
carrier is a vegetable oil.
14. The method of claim 9 wherein said composition is formulated as
a paste, cream, ointment, gel, lotion or unguent.
15. The stereospecific optically pure trans (E) isomer of coenzyme
Q 10 produced by the process of claim 1.
Description
BACKGROUND OF THE INVENTION
[0001] The invention relates to an improved process for the
stereospecific synthesis of Coenzyme Q10, ubiquinone. The present
invention also relates to the therapeutically useful optically pure
isomers of Coenzyme 10 and refers to new pharmaceutical
compositions which contain the optically pure isomers of coenzyme
Q10 dissolved or suspended in a suitable vehicle which are useful
for example in preventing anoxic tissular damage, particularly in
the myocardium. Previous procedures for ubiquinone isolation bad
several drawbacks; many steps were involved, the yields were low,
the intermediates were difficult to purify, overall costs were
high, and the final products were obtained as mixtures of isomers,
cis(Z) and trans (E).
[0002] Coenzyme Q gives reference to a series of quinones which are
widely distributed in animals, plants and microorganisms. These
quinones have been shown to function in biological electron
transport systems which are responsible for energy conversion
within living cells. In structure, the coenzyme Q group closely
resembles the members of the vitamin K group and the
tocopherylquinones, which are derived from tocopherols (vitamin E),
in that they all possess a quinone ring attached to a long
hydrocarbon tail. The quinones of the coenzyme Q series which are
found in various biological species differ only slightly in
chemical structure and form a group of related,
2-3-dimethoxy-5-methyl-benzoquinones with a polyisoprenoid side
chain in the 6-position which varies in length from 30 to 50 carbon
atoms. Since each isoprenoid unit in the chain contains five carbon
atoms, the number of isoprenoid units in the side chain varies from
6 to 10. The different numbers of the groups have been designated
by a subscript following the Q to denote the number of isoprenoid
units in the side chain, as in Q10. Difference in properties are
due to the difference in length of the side chain. The members of
the group known to occur naturally are Q6 through Q10. Coenyme Q
functions as an agent for carrying out oxidation and reduction
within cells. Its primary site of function is in the terminal
electron transport system where it acts as an electron or hydrogen
carrier between the flavoproteins (which catalyze the oxidation of
succinate and reduced pyridine nucleotides) and the cytochromes.
This process, is carried out in the mitochondria of cells of higher
organisms. Certain bacteria and lower organisms do not contain any
coenzyme Q. It has been shown that many of these organisms contain
vitamin K, instead and that this quinone functions in electron
transport in much the same way as coenzyme Q. Similarly, plant
chloroplasts do not contain coenzyme Q, but do contain
plastoquinones, which are structurally related to coenzyme Q.
Plastoquinone functions in the electron transport process involved
in photosynthesis. In some organisms, coenzyme Q is present
together with other quinones, such as vitamin K,
tocopherylquinones, and plastoquinones; and each type of quinone
can carry out different parts of the electron transport
functions.
[0003] Coenzyme Q 10, is a ubiquinone. Ubiquinones are a class of
lipid soluble benzoquinones that are involved in mitochondrial
electron transport and are essential electron and proton carriers
that function in the production of biochemical energy in all cells
of aerobic organisms; participating in the transport of electrons
from organic substrates to oxygen in the respiratory chain of
mitochondria. In addition, coenzyme Q10 has antioxidant and
membrane stabilizing properties that serve to prevent cellular
damage resulting from normal metabolic processes. It plays an
important role as an antioxidant to neutralize potentially damaging
free radicals created in part by the energy-generating process. As
an energy carrier, coenzyme Q10 is continually going through an
oxidation reduction cycle. As each coenzyme Q10 molecule accepts
electrons, it is reduced, when it gives up electrons, it becomes
oxidized again. In coenzyme Q10's reduced form (ubiquinol), the
coenzyme Q 10 molecule holds electrons loosely and will quite
easily give up one or two electrons to neutralize free radicals. In
its electron rich reduced form, coenzyme Q10 is as potent an
antioxidant as vitamin E. Coenzyme Q10's main role as an
antioxidant is in the mitochondria where it first participates in
the process by which free radicals are generated and then helps to
quench the extra free radicals that threaten cellular components
such as DNA, RNA, and cell membranes. One of coenzyme Q10's key
antioxidant actions is within the cell membrane, where it counters
the oxidative attack of polyunsaturated lipids (lipid
peroxidation), which causes damage in a self-propagating,
destructive chain reaction that ultimately results in membrane
degeneration leading to cell death.
[0004] In mammamlian tissue the quinone ring of coenzyme Q10 is
synthesized from the amino acids, tyrosine and phenylalanine and
the polyprenyl side chain is synthesized from acetyl-CoA. The
number of isoprene units depends on the species, the most common
form in mammals contains ten isoprene units. Coenzyme Q10
participates in the transport of electrons from organic substrates
to oxygen in the respiratory chain of the mitochondria. During this
process ubiquinone is reduced to a free radical semiquinone by the
uptake of a single electron. Reduction of this enzyme-bound
intermediate by a second electron yields ubiquinol. This a
reversible reducible process.
[0005] Ubiquinone has a characteristic light absorption band at 270
to 290 nm, which disappears when it is reduced to its quinol form;
this spectral change is used to measure oxidation and reduction of
ubiquinone.
[0006] The structure of coenzyme Q10 consists of a quinone ring
attached to an isoprene side chain. It contains 82.08% carbon,
10.51% hydrogen and 7.41% oxygen. Its has a molecular weight of
863.37 and a formula of C.sub.59H.sub.90O.sub.4. The oxidized,
intermediate and reduced forms of coenzyme Q10 are shown in the
following drawings: 1
[0007] Coenzyme Q10 is used extensively as a nutritional supplement
as well as a pharmacological active agent. It has wide use and
acceptance in the treatment of cardiovascular disease in
traditional as well as alternative medicine. It is used
successfully in treating ischemic heart disease, chronic heart
failure, toxin induced cardiomyopathy, hypertension and
hyperlipidemia. Endogenous coenzyme Q10 functions as an essential
cofactor in many metabolic pathways. Its action as an additional
pharmacological agent in treatment of such cardiovascular disease
processes may be to improve function of the involved tissues that
are ischemic or pathologically altered by providing an increased
energy source, by acting as a free radical scavenger and/or
membrane stabilizer. In addition, coenzyme Q10 is found in high
concentrations in healthy hearts and at low levels in people with
congestive failure leading to the suggestion that supplementation
with the coenzyme would be of help in the treatment of heart
disease. It is theorized that Coenzyme Q10 might work in the heart
in two ways; as an antioxidant to help thwart damage from free
radicals that contribute to arterial blockage, and to help boost
heart muscle action by improving energy efficiency. Additionally
coenzyme Q10 may boost the effects of vitamin E, also a potent
antioxidant with some potential beneficial heart effects.
[0008] Coenzyme Q10 has been used in the treatment of slow muscle
degeneration (dystrophy or atrophy) and the accompanying cardiac
complications typically found in these patients
[0009] In addition to its helper role in the release of energy,
Coenzyme Q10 serves as an antioxidant, neutralizing free radicals
that cause potentially irreversible damage to cells, tissues, and
organs. Coenzyme Q10 is also believed to strengthen the immune
system, so as to provide antibacterial and antiviral activity
(including HIV), to increase antibody production and to induce the
immune system to produce a greater number of immune acting cells.
Among the increasing number of pharmacological uses ascribed to
coenzyme Q10 are anticancer (in particular breast cancer) activity,
in the treatment of periodontal disease, diabetes, Parkinson's,
Alzheimer's, Huntington's disease and to help counteract the aging
process.
[0010] The rationale for its effectiveness in relieving certain
brain disorders is that coenzyme Q10 temporarily restores
mitochondrial activity in cells. There is evidence that Parkinson's
disease, Huntington's disease and some other neurological diseases
may impair the mitochondria throughout the body, but particularly
in nerve and brain cells. In that case coenzyme Q10 might slow the
progression of these diseases. As a potent. antioxidant, coenzyme
Q10 might also help prevent the cell death that occurs in these
diseases by blocking the buildup of toxic substances. It has also
been shown, that coenzyme Q10 lowers levels of lactate in the
brains of people with Huntington's disease. Increased lactate
suggests a problem with energy metabolism in brain cells.
[0011] Free radical damage is thought to be an important
contributor to the body wide deterioration that accompanies aging.
Laboratory evidence suggests that supplementation with Coenzyme Q10
can at least partially protect against such damage.
[0012] It is clear from the literature that activity of coenzyme
Q10 is strictly connected with the tissular respiratory
processes.
[0013] A wide bibliography points out its ability to solve or
prevent anoxic tissular damages, particularly in the
myocardium.
[0014] Other positive effects have been obtained by means of
coenzyme Q10 in the treatment of arterial hypertension, of muscular
dystrophy, of periodontopathies, of penfigus and of lichen
planus.
[0015] In all such pathological conditions, it was also noticed
that the administration of coenzyme Q10 led to a normalization of
tissular concentrations of this enzyme, otherwise scarce.
[0016] Coenzyme Q10 is not toxic (there are no reported side
effects), no known medical conditions preclude it use. It is
generally employed as a supplement,.rather than a replacement for
standard medical treatment. No known drug interactions have been
reported. Daily oral doses vary from 5-10 mg/dosage (15 to 30 mg
pro die) to 50-100 mg(dosage (100-200 mg pro die) The
administration of even higher dosages up to 400 mg pro die give
satisfactory clinical results but it increases sometimes, the pro
die effects of the drug. The larger amount usually being given as
multiple doses.
[0017] There seems to be no limiting factor as to how long the
coenzyme Q10 may be taken; individuals have used it continuously
for years.
DESCRIPTION OF THE INVENTION
[0018] The invention relates to a process for the stereospecific
synthesis of coenzyme Q10, ubiquinone. In accordance with one
embodiment of the invention, the synthetic process utilizes
solanesol sourced from tobacco waste as the starting material. In
accordance with another embodiment of the invention the synthetic
process utilizes potato leaves as the starting material.
[0019] When solanesol derived from tobacco and potato leaves is
used as the starting material, the method of production of coenzyme
Q 10 involves a semi synthesis, if geraniol is used as the starting
material what is involved is a total synthesis. The two methods of
synthesis as hereinafter set forth produce high yields of
isomerically pure coenzyme Q10. Rather than using huge quantities
of costly tobacco leaves in the solanesol process, it has been
found that tobacco dust, a waste product of the tobacco industry
also can effectively be used as the starting material.
[0020] The previous methods of production of Coenzyme Q 10 had many
disadvantages. The procedures were lengthy and involved many steps,
the resulting yields were low, the intermediates were difficult to
purify, overall costs were high and the final products were
obtained as mixtures of isomers, cis(Z) and trans(E).
[0021] In addition to alleviating the described disadvantages, the
processes of the present invention have been found to be
stereospecific (selective) producing exclusively the desired all
trans(E) isomers.
[0022] The following example will serve to illustrate the
invention, it being understood that the same is not to be construed
in limitation thereof.
EXAMPLE
[0023] Step 1: Solanesol.
[0024] One kg of tobacco dust is shaken with 4 l of hexane for 1.5
hours. The solid is separated by filtration and extracted with a
total of 2.4 l of hexane. The combined extracts are evaporated. To
the resulting residue are added 140 ml of 2 N KOH in ethanol and 2
g of pyrogallol. The obtained mixture is heated for 1 hour (reflux)
under N.sub.2. It is then rapidly cooled, 400 mg of a 9:1 mixture
of ethanol and water is added and this mixture extracted with
hexane (4.times.400 ml). The extract is dried with sodium sulfate
and chromatographed on a column of 300 g of alumina. After elution
with a 9:1 mixture of hexane/ether and evaporation, 5 g of
solanesol are obtained.
[0025] Step 2: Solanesylacetone.
[0026] To a solution of 10 g of solanesol obtained in Step 1 in a
mixture of 9.5 ml of anhydrous hexane and 13.5 ml anhydrous ether,
at 0-5 degree C., there are added 1.3 ml of pyridine. Thereafter
over a period of 35 minutes, 1.8 ml of phosphorous tribromide in 13
ml of hexane are added. The resulting mixture is stirred, at 0-5
degrees C. for 3 hours and then added to ice water, and then
stirred for an additional 10 minutes. The organic phase is
separated off, and the aqueous layer extracted with ether, washed
with 5% sodium bicarbonate water, and then dried with magnesium
sulfate. After evaporation, 11 g of solanesol bromide are obtained.
2.5 g of ethyl acetoacetate are added to the solanesol bromide
followed by the addition of a solution of 0.4 g of sodium in 18 ml
absolute alcohol, over a period of 20 minutes while maintaining a
temperature of 10 degrees C. The resulting reaction mass is kept at
20 degrees C. for 12 hours. It is then heated to 80 degrees C., and
2.5 ml of 10% NaOH are added over 1 hour. The mixture is then
stirred at 80 degrees C. for 4 hours, poured into ice water and
extracted with ether, The ether solution is washed with water and
dried with magnesium sulfate and evaporated. The yield is 8 g of
solanesylacetone obtained as a solid.
[0027] The chemical reactions can be seen in the following
formulas: 2
[0028] Step 3:--Isodecaprenol.
[0029] To 1.4 g of magnesium in 18 ml anhydrous THF (under N.sub.2)
a small crystal of iodine, a drop of methyl iodide and 0.1 ml of
vinyl bromide are added. After the onset of the exothermic
reaction, 4 ml vinyl bromide in 9 ml THF are gradually added while
the temperature is maintained at about 50 degrees C. The mixture is
stirred at 50-60 degrees C. for 1 hour to complete the formation of
vinyl-magnesium bromide. The mixture is then cooled to 0-5 degrees
C. and 8 g of solanesylacetone in 32 ml THF are added over a period
of 10 minutes. The mixture is left at 20 degrees C. for 3 hours
after which it is cooled to 0-5 degrees C. A solution of 4.2 g
ammonium chloride in 10 ml of water is added, and mixture is
stirred for 10 minutes and is then extracted with ether, washed
with water and dried with magnesium sulfate. After evaporation, 9 g
of isodecaprenol were obtained as a colorless, waxy substance.
[0030] Step 4:--2, 3, 6-tribromo-4-methylphenol-9108 g (1 mol)
p-cresol is dissolved in 120 l of chloroform containing 4 g iron
powder and 500 g bromine are then introduced drop wise over a
period of 5 hours at room temperature. The resulting solution is
stirred for 48 hours and then filtered, washed with dilute sodium
sulfite and dried with magnesium sulfate. The solvent is evaporated
and the residue recrystallized from hexane. A yield of 265 g is
obtained. 3
[0031] Step 5:--2,3,4,5-tetramethoxytoluene.
[0032] Sodium (23 g) is dissolved in 400 ml of methanol. To this
9.3 g DME and 50 ml dimethyl carbonate are added to the resulting
solution. Most of the methanol (300 ml) is then removed by
distillation; 915 g of copper cyanide are added and a solution of
34.4 g 2,3,6tribromo-4-methylphenol in 100 ml DME prepared and is
added drop wise over a period of 3 hours, with stirring, while
maintaining the temperature at 80 degrees C. The mixture is stirred
an additional 5 hours while continuing to maintain the 80 degrees
C. temperature. To this mixture, 800 ml of water is added, the
mixture is cooled to 50 degrees C. 100 ml Dimethyl sulfate is then
added in drop wise fashion. The mixture is then stirred at room
temperature for 2 hours and concentrated. 250 Ml aqueous ammonium
hydroxide are then added. The mixture is extracted with
methylchloride and the organic extract washed with dilute HCl and
water and dried over magnesium sulfate. The solvent is evaporated
off leaving 20 g of pure product; bp 100 C. (0.1 mm). 4
[0033] Step 6:--2,3 dimethoxy-5-methylhydroquinone.
[0034] 33.4 g of Pyridine-2,6 dicarboxylate was added to a cold, 0
degree C., solution of 2,3,4,5 tetramethoxytoluene (17 g) in 400 ml
of acetonitrile/water (7:3). A cold solution, 0 degree C., of ceric
ammonium nitrate, 110 g in 400 ml of 1:1 acetonitrile/water is
slowly added over 20 minutes, and the mixture stirred for 20
minutes at 0 degree C. and for 10 minutes at room temperature. The
reaction mixture is poured into 400 ml of water and extracted with
methyl chloride. The organic layer is dried over magnesium sulfate,
evaporated and chromatographed on silica gel, (hexane/ethylacetate
20:1, yielding 14 g of red crystals, mp 59 degrees C. 5
[0035] Step 7:--Ubiquinone Coenyme Q10.
[0036] Equimolar portions of portions of the products obtained in
step 3 (isodecaprenol)and 6 (2,3 dimethoxy-5-methyl hydroquinone)
are stirred at 43 degrees C. for 10 minutes in hexane. A 2.5%
sodiumbisulfite solution is added, and the hexane layer separated,
dried over magnesium sulfate, concentrated and chromatographed on
silica gel (hexane/ether 10:1) to give ubiquinone as a yellow
solid. 6
[0037] The administration of optically pure Coenzyme Q10 can be
oral, parenteral or topically, in the latter case for the treatment
of diseases of skin and mucous membranes. Oral administration is
favored over parenteral administration due to the very low
solubility of coenzyme Q10 in excipients compatible with is
parenteral administration. Oral administration has proved
particularly useful in the treatment of diseases affecting
metabolically very active organs, whereas coenzyme Q 10 if
administered orally proves to be substantially ineffective at the
cutaneous level. Accordingly the concentration of Coenzyme Q 10 has
to be increased for topical administration directly to impaired or
damaged tissue.
[0038] The oral form of administration can be as pills, tablets,
capsules or liquid preparations in each case formulated in the
conventional manner with suitable carriers and formulation aids.
The formulations are prepared to deliver 5-100 mg per dosage unit
and in some instances up to 200 mg per dosage units of the
optically pure coenzyme Q 10.
[0039] The compositions for topical administration can be prepared
by dissolving or suspending coenzyme Q 10 in vegetable oils such as
corn oil, canola oil, or soy bean oil, lecithin, glycerol,
glycerylfurole, Tween 80 or other derivatives, suspending agents or
diluents. After the addition of suitable carriers and formulation
aids to such solutions or suspensions, the compositions can be
forwarded as pastes, creams, ointments, gels, lotions,
unguents.
[0040] The compositions for topical application contain the
optically pure coenzyme 10 as the active principal in amounts from
0.1 to 10%, preferably from 0.25 to 1%. The topical compositions
can also be used for cosmethological purposes. In such a case, the
content of coenzyme Q 10 can be lower than the limits
aforementioned being preferably from 0.0001 to 0.1%.
[0041] The compositions in any application form may also contain
other topically active components beside the active principle
(optically active coenzyme Q 10).
* * * * *