Note: Descriptions are shown in the official language in which they were submitted.
CA 02333752 2000-11-30
' WO 99/62524 PCT/US98/11748
AMINOSUGAR, GLYCOSAMINOGLYCAN, AND S-ADENOSYLMETHIONINE
COMPOSITION FOR THE TREATMENT AND
REPAIR OF CONNECTIVE TISSUE
to
BACKGROUND OF THE INVENTION
Cross-Reference to Related Application
The present application is a continuation-in-part of co-pending United States
Patent Application Serial No. 08/779,996, filed December 23, 1996, the
disclosure of
which is incorporated by reference herein in its entirety.
1. Field of the Invention
The present invention relates to compositions for the repair and reduction of
inflammation of connective tissue in humans and animals and, in particular, to
2 0 compositions capable of promoting anti-inflammation, chondroprotection,
chondromodulation, chondrostabilization, chondrometabolization and the repair
and
replacement of human and animal connective tissue.
2. Background of the Invention
The connective tissues of humans and animals are constantly subjected to
stresses
2 5 and strains from mechanical forces and from diseases that can result in
afflictions, such as
arthritis, joint inflammation and stiffness. Indeed, connective tissue
afflictions are quite
common, presently affecting millions of Americans. Further, such afflictions
can be not
only painful but, in their extreme, debilitating.
The treatment of connective tissue afflictions can be quite problematic. A
simple
3 o decrease in the stress to which the connective tissue is subjected is
often not an option,
- 1 -
CA 02333752 2000-11-30
WO 99/62524 PCT/US98/11748
especially in the case of athletes and animals such as race horses.
Consequently,
treatment is often directed at controlling the symptoms of the afflictions and
not their
causes, regardless of the stage of the degenerative process.
Presently, steroids, such as corticosteroids and NSAIDs, are widely used for
the
treatment of these ailments. [Vidal, et al., Pharmocol. Res. Commun., 10;557-
569
(1978)]. However, drugs such as these, which inhibit the body's own natural
healing
processes, may lead to further deterioration of the connective tissue.
Connective tissue, for example articular cartilage, is naturally equipped to
repair
itself by manufacturing and remodeling prodigious amounts of collagen (a chief
1 o component of connective tissue such as cartilage) and proteoglycans (PGs)
(the other
major component of connective tissue such as cartilage). This ongoing process
is placed
under stress when an injury occurs. In such cases, the production of
connective tissue
matrix (collagen and PGs) can double or triple over normal levels, thereby
increasing the
demand for the building blocks of both collagens and proteoglycans.
The building blocks for collagen are amino acids, especially proline, glycine
and
lysine. PGs are large and complex macromolecules comprised mainly of long
chains of
modified sugars called glycosaminoglycans (GAGs) or mucopolysaccharides. The
terms
GAGS and mucopolysaccharides are understood in the art to be interchangeable.
PGs
provide the framework for collagen formation and also hold water to give
flexibility,
2 0 resiliency and resistance to compression.
Like almost every biosynthetic pathway in the body, the pathways by which both
collagen and GAG form single molecule precursors are quite long. As is also
characteristic of other biosynthetic pathways, the pathways by which collagen
and GAGS
are produced include what is called a rate-limiting step -- that is, one
highly regulated
2 5 control point beyond which there is a commitment to finish. The presence
of such rate-
- 2 -
CA 02333752 2000-11-30
WO 99/62524 PCT/US98111748
limiting steps permits complicated biosynthetic processes to be more easily
and
efficiently controlled by permitting the organism to focus on one point. For
example, if
conditions demand production and all the requisite raw materials are in place,
then
stimulation of the rate-limiting step will cause the end product to be
produced. To stop or
slow production, the organism needs simply to regulate the rate-limiting step.
In the production of PGs, the rate-limiting step is the conversion of glucose
to
glucosamine for the production of GAGS. Glucosamine, an aminosugar, is the key
precursor to all the various modified sugars found in GAGs, including
glucosamine
sulfate, galactosamine, N-acetylglucosamine, etc. Glucosamine also makes up to
50% of
hyaluronic acid -- the backbone of PGs -- on which other GAGs, like
chondroitin sulfate
are added. The GAGS are then used to build PGs and, eventually, connective
tissue.
Once glucosamine is formed, there is no turning away from the synthesis of GAG
polymers.
Glucosamine has been shown to be rapidly absorbed into humans and animals
after oral administration. A significant portion of the ingested glucosamine
localizes to
cartilage and joint tissues, where it remains for long periods. This indicates
that oral
administration of glucosamine reaches connective tissues, where glucosamine is
incorporated into newly-synthesized connective tissue.
Glycosaminoglycans and collagen are the chief structural elements of all
2 0 connective tissues. Their synthesis is essential for proper maintenance
and repair of
connective tissues. In vitro, the introduction of glucosamine has been
demonstrated to
increase the synthesis of collagen and glycosaminoglycans in fibroblasts,
which is the
first step in repair of connective tissues. In vivo, topical application of
glucosamine has
enhanced wound healing. Glucosamine has also exhibited reproducible
improvement in
2 5 symptoms and cartilage integrity in humans with osteoarthritis. [L. Bucci,
Nutritional
- 3 -
CA 02333752 2000-11-30
WO 99/62524 PCT/US98/11748
Supplement Advisor, (July 1992)].
The pathway fox the production of proteoglycans may be briefly described as
follows. Glucosamine is the main building block of connective tissue and may
be
provided either through the enzymatic conversion of glucose or through diet or
external
administration (see FIG. 1 ). Glucosamine may be converted into the other main
component of connective tissue, namely PGs, upon incorporation of glucosamine
into
GAGS (see FIG. 2).
More specifically, GAGS are large complexes of polysaccharide chains
associated
with a small amount of protein. These compounds have the ability to bind large
amounts
of water, thereby producing a gel-like matrix that forms the body's ground
substance.
GAGS stabilize and support cellular and fibrous components of tissue while
maintaining
the water and salt balance of the body. The combination of insoluble protein
and the
ground substance forms connective tissue. For example, cartilage is rich in
ground
substance while tendon is composed primarily of fibers.
GAGs are long chains composed of repeating disaccharide units of
monosaccharides (aminosugar-acidic sugar repeating units). The aminosugar is
typically
glucosamine or galactosamine. The aminosugar may also be sulfated. The acidic
sugar
may be D-glucuronic acid or L-iduronic acid. GAGS, with the exception of
hyaluronic
acid, are covalently bound to a protein, forming proteoglycan monomers. These
PGs
2 0 consist of a core protein to which linear carbohydrate chains formed of
monosaccharides
are attached. In cartilage proteoglycan, the species of GAGs include
chondroitin sulfate
and keratin sulfate. The proteoglycan monomers then associate with a molecule
of
hyaluronic acid to form PG aggregates. The association of the core protein to
hyaluronic
acid is stabilized by link proteins.
2 5 The polysaccharide chains are elongated by the sequential addition of
acidic
- 4 -
CA 02333752 2000-11-30
WO 99/62524 PCT/US98/11748
sugars and aminosugars, and the addition is catalyzed by a family of
transferases.
Aminosugars, such as glucosamine, are synthesized through a series of
enzymatic
reactions that convert glucose to glucosamine, or alternatively may be
provided through
the diet. The glucosamine is then incorporated into the GAGS as described
above. Acidic
sugars may be provided through the diet, may be obtained through degradation
of GAGs
by degradative enzymes, or produced through the uronic acid pathway.
Since repeating disaccharide units contain one aminosugar (such as
glucosamine),
it is clear that the presence of an aminosugar in the production of connective
tissue is
important. Glucosamine is, by far, the more important ingredient in the
production of
connective tissue since it is the essential building block of GAGS. See FIG 1.
All GAGs
contain hexosamine or uronic acid derivative products of the glucose pathway
and from
exogenous glucosamine, for example:
Hyaluronic acid Glucosamine + Glucuronic Acid
Keretan-Sulfate Glucosamine + Galactose
Chondroitin Glucuronic Acid + Galactosamine
Sulfate
Heparin Sulfate Glucosamine + Glucuronic or Iduronic Acid
Heparan Sulfate Glucosamine + Glucuronic or iduronic Acid
Dermatin Sulfate Iduronic Acid + Galactosamine
Chondroitin sulfate is a GAG that provides a further substrate
for the synthesis of
2 0 the proteoglycans.The provision of the chondroitin in its salt
(sulfate) form facilitates its
delivery and uptake by the humans and animals in the production of connective
tissue. In
addition, the sulfate portion of chondroitin sulfate is available for use in
catalyzing the
conversion of glucosamine to GAGs. Fragments of GAGs, including chondroitin
sulfate,
may also be used to provide a substrate for synthesis of proteoglycans since
the assembly
2 5 of PG occurs in the extracellular space.
In addition, chondroitin sulfate has been shown to have cardiovascular health
benefits. [Morrison et al., Coronary Heart Disease and the Mucopolysaccharides
- 5 -
CA 02333752 2000-11-30
WO 99/62524 PCT/US98/11748
(Glycosaminoglycans), pp. 109-127 (1973)]. Thus, the preferred form of
glycosaminoglycan included in the compositions of the present invention is
chondroitin
sulfate or fragments thereof.
Chondroitin may be more efficacious than glucosamine for injury
rehabilitation.
[Christensen, Chiropractic Products, pp. 100-102 (April 1993)]. An evaluation
of
glucosamine versus chondroitin for treatment of osteoarthritis has been
conducted and
concludes, contrary to Christensen, that glucosamine is preferred. [Murray,
MPI's
Dynamic Chiropractic, pp. 8-10 (September 12, 1993)]. Neither reference
teaches or
suggests combining the materials. Bucci (Townsend Letter for Doctors, pp. 52-
54,
1 o January 1994), discloses the combination of glucosamine and chondroitin
for treatment of
osteoarthritis. Bucci has acknowledged that this combination was personally
disclosed to
him by one of the present inventors.
Chondroitin sulfate also acts to inhibit the degradative enzymes that break
down
connective tissue. In so doing, chondroitin sulfate promotes the maintenance
of healthy
connective tissues. When combined with glucosamine, which functions primarily
as a
building block fox the synthesis of connective tissue, chondroitin sulfate
works in concert
with the glucosamine but may work in a different fashion. The ability of
chondroitin
sulfate to block degradation is one of its important functions.
S-Adenosylmethionine (SAM) is a significant physiologic compound which is
2 o present throughout body tissue and takes part in a number of biologic
reactions as a
methyl group donor or as an enzymatic activator during the synthesis and
metabolism of
hormones, neurotransmitters, nucleic acids, phospholipids, and proteins. SAM
may be
second only to adenosine triphosphate (ATP) in the variety of reactions in
which it is a
cofactor. SAM is metabolized via three metabolic pathways of transmethylation,
transsulfuration, and aminopropylation. [Stramentinoli, Am. J. Med., 83(SA):35-
42
- 6 -
CA 02333752 2000-11-30
WO 99/62524 PCT/US98/11748
(1987)]. In higher organisms, SAM plays a significant role in transmethylation
processes
with more than 40 anabolic or catabolic reactions involving the transfer of
the methyl
group of SAM to substrates such as nucleic acids, proteins, and lipids, among
others.
Also, the release of the methyl group from SAM is the start of a
"transsulfuration"
pathway that produces all endogenous sulfur compounds. After donating its
methyl
group, SAM is converted into S-adenosylhomocysteine, which in turn is
hydrolyzed to
adenosine and homocysteine. The amino acid cysteine may then be produced from
the
homocysteine. The cysteine thus produced may exert a reducing effect by itself
or as an
active part of glutathione, which is a main cell anti-oxidant. [Stramentinoli,
cited above].
1 o SAM has been used to treat various disorders. In various forms of liver
disease,
SAM acts as an anticholestatic agent. [Adachi et al., Japan Arch. Inter. Med.,
33:185-192
(1986)]. SAM has also been administered as an antidepressant for use in the
management
of psychiatric disorders [Caruso et al., Lancet, l: 904 (1984)], and as an
anti-
inflammatory compound in the management of osteoarthritis [Domljan et al.,
Int. J. Clin.
Pharm. Toxicol., 27(7):329-333 (1989)].
Low levels of SAM are believed to play a role in reducing the risk of certain
cancers. [Feo et al., Carcino eg nesis, 6:1713-20 (1985)]. In addition, the
administration of
SAM has also been associated with a fall in the amount of early reversible
nodules and
the prevention of the development of late pre-neoplastic lesions and
hepatocellular
2 0 carcinomas. [Garcea et al., Carcino eg nesis, 8:653-58 (1987)].
Unfortunately, SAM ep r se is unstable due to its high reactivity. The
relatively
recent synthesis of stable salts, however, has made SAM available for research
and
therapeutic use. [See, e.g., U.S. Patent Nos. 4,990,606 and 5,102,791].
SAM has been used outside of the United States in a number of clinical trials
2 5 concerning the treatment of osteoarthritis. While used in these trials
primarily as an
CA 02333752 2000-11-30
WO 99/62524 PCT/US98/11748
analgesic and replacement for NSAID therapy, SAM is a precursor of polyamines.
In
addition to their analgesic and anti-inflammatory properties, and their
ability to scavenge
free radicals, polyamines may stabilize the polyanionic macromolecules of
proteoglycans.
[Schumacher, Am. J. Med., $3(SA):2 (1987)].
SAM may also function as a source of endogenous sulfur, which will increase
sulfation of GAGs to be incorporated in proteoglycans. The inclusion of SAM is
particularly beneficial in instances of subclinical deficiencies of SAM,
occurring
especially in elderly populations with higher risk of osteoarthritis [Frezza
et al.,
Gastroenterol., 99:211-215 {1990)]. The supplementation of SAM may aid in
instances
of SAM deficiency where the ability of the body to sulfate GAGS may be
compromised.
In addition, a number of metabolites of SAM aid in the repair of connective
tissue,
including glutathione, polyamines, methylthioadenosine, and adenosine.
Glutathione
works as a scavenger of oxygen-related products [Shumacher, Am. J. Med.,
83(Supp
Sa):1-4 (1987); Matthew & Lewis, Pharmacol. (Life Sci. Adv.), 9:145-152
(1990); Szabo
et al., Science, 214:200-202 ( 1981 )] and thus has an anti-inflammatory
effect.
Polyamines, including spermine, spermidine, and putrescine, stabilize
polyanionic
macromolecules of proteoglycans [Schumacher, cited above; Conroy et al.,
Biochem. J.,
162:347-350 (1977)] and thus protect proteolytic and glycolytic enzymes. These
polyamines also have an anti-inflammatory effect [Bird et al., Agents Actions,
13:342-
347 (1983); Oyangui, Agents Actions, 14:228-237 (1984)], probably as a
scavenger of
oxygen-related products [Kafy et al., Ag-ents Actions, 18:555-559 (1986);
Matthews &
Lewis, cited above], and have an analgesic effect [Bird et al., cited above;
Oyangui, cited
above]. The SAM metabolite methylthioadenosine has a pronounced anti-
inflammatory
effect [Matthews & Lewis, 1990] while adenosine has a more modest anti-
inflammatory
effect [Matthews & Lewis, 1990].
_ g _
CA 02333752 2000-11-30
WO 99/62524 PCT/US98/11748
Studies have shown that some forms of exogenous SAM are stable in digestive
juices when given orally. [Stramentinoli et al., cited above; Vendemiale et
al., Scand. J.
Gastroenterol., 24:407-415 (1989)]. The metabolism of exogenous SAM appears to
follow known pathways of endogenous SAM metabolism. [Kaye et al., Dru s,
40(Suppl.
3):124-138 (1990)). In humans, oral SAM was tolerated to the same extent as
placebo
with very mild nonspecific side effects. [Schumacher, cited above; Frezza et
al., cited
above].
Manganese plays a role in the synthesis of GAGs, collagen and glycoproteins
which are important constituents of cartilage and bone. Manganese is important
for
enzyme activity of glycosyltransferases. This family of enzymes is responsible
for
linking sugars together into glycosaminoglycans, adding sugars to other
glycoproteins,
adding sulfate to aminosugars, converting sugars into other modified sugars,
and adding
sugars to lipids. The enzymatic functions of glycosyltransferases are
important in
glycosaminoglycan synthesis (hyaluronic acid, chondroitin sulfate, keratan
sulfate,
heparin sulfate and dermatin sulfate, etc.), collagen synthesis, and in the
functions of
many other glycoproteins and glycolipids.
Manganese also plays a role in the synthesis of glycosaminoglycans and
glycoproteins, which are important constituents of cartilage and bone. Many
reproductive
problems in horses and skeletal abnormalities in foals have been ascribed to
manganese
2 0 deficiency. [Current Therapy in Eguine Medicine, 2:402-403 (1987)).
Manganese deficiency leads to abnormal bone growth, swollen and enlarged
joints, and slipped tendons in humans and animals. In humans, manganese
deficiencies
are also associated with bone loss and arthritis. Levels of all
glycosaminoglycans are
decreased in connective tissues during manganese deficiencies, with
chondroitin sulfates
2 5 being most depleted. Manganese-deficient organisms quickly normalize
_ g _
CA 02333752 2000-11-30
WO 99/62524 PCT/US98/11748
glycosaminoglycans and collagen synthesis when manganese is replenished.
Approximately 40% of dietary manganese is absorbed by the body tissue. Storage
of manganese in the body is minimal -- a mere 12 to 20 mg is present in the
body at any
one time. Large amounts of calcium and phosphorus in the intestine are also
known to
interfere with manganese absorption. The richest dietary sources are the foods
least
consumed by the general public, such as whole grain cereals and breads, dried
peas, beans
and nuts. The ascorbate form of manganese is preferred due to the high
bioavailability
and the need for vitamin C (ascorbic acid) for collagen production. Vitamin C
also
enhances manganese uptake by the body.
Other optional ingredients in the compositions of the present invention are
methyl
donors or methyl donor cofactors, such as vitamins B 12 and B6, folic acid,
dimethylglycine, and trimethylglycine. These ingredients augment the function
of SAM
in that they are cofactors in methylation. In addition, these compounds are
likely to be
lacking in patients suffering from connective tissue disorders. For example,
it is
estimated that 12% of the elderly population in the United States suffers from
a vitamin
B 12 deficiency, a group more likely to suffer from connective tissue
disorders.
An adequate amount of vitamin B 12, for example, has an important
environmental influence on the accumulation of homocysteine that results from
the
metabolism of SAM. In other words, methyl donors or methyl donor cofactors,
such as
2 0 vitamin B 12 and the others listed in the preceding paragraph, can reduce
levels of
homocysteine when administered either alone or in combination.
Vitamin B12 is generally known to function as a coenzyme in biochemical
reactions such as the synthesis of proprionic acid and of methionine. Recent
evidence
suggests that the elevated levels of plasma homocysteine increase the risk of
occlusive
2 5 vascular disease. Adequate amounts of vitamin B I2 are considered the most
important
- 10 -
CA 02333752 2000-11-30
WO 99/62524 PCT/US98/11748
environmental influence on the accumulation of unnecessary homocysteine.
[Joosten et
al., Am. J. Clin. Nutr., 58(4): 468-76 (1993)]. In addition, it is also
understood that
vitamin B 12 may play a role in the methylation of selenium. [Chen and
Whanger, Tox.
and Appl. Pharm., 118:65-72 (1993)]. Specifically, increased levels of vitamin
B12
significantly contribute to selenium methylation and might decrease overall
selenium
toxicity by preventing its accumulation in tissues. [Chen and Whanger, cited
above).
3. Description of Background Art
Several disclosures suggest provide exogenous quantities of glucosamine in
order
to bypass the rate-limiting step of the conversion of glucose to glucosamine
in those
l0 pathways that produce PGs. For example, the intravenous administration of
glucosamine
(a precursor of the GAGS) and derivatives thereof has been disclosed in United
States
Patent No. 3,232,836, issued to Carlozzi et al., for assisting in the healing
of wounds on
the surface of the body. In United States Patent No. 3,682,076, issued to
Rovati, the use
of glucosamine and salts thereof is disclosed for the treatment of arthritic
conditions.
Finally, the use of glucosamine salts has also been disclosed for the
treatment of
inflammatory diseases of the gastrointestinal tract in United States Patent
No. 4,006,224
issued to Prudden. In vitro, glucosamine increases synthesis of collagen and
glycosaminoglycans, the first step in repair of connective tissues, in
fibroblasts. In vivo,
topical application of glucosamine has enhanced wound healing.
2 0 Several disclosures also suggest going one step further in bypassing the
glucose-
to-glucosamine rate-limiting step, by providing exogenous quantities of
various of the
modified sugars found in the GAGs for producing proteoglycans. For example, in
United
States Patent No. 3,6797,652 issued to Rovati et al., the use of N-
acetylglucosamine is
disclosed for treating degenerative afflictions of the joints.
2 5 In still other disclosures of which we are aware, it has been taught to go
still one
- 11 -
CA 02333752 2000-11-30
- WO 99/62524 PCT/US98111748
step further in bypassing the glucose-to-glucosamine rate-limiting step by
providing
exogenous quantities of the GAGS themselves (with and without various of the
modified
sugars). For example, in United States Patent No. 3,371,012 issued to
Furuhashi, a
preservative is disclosed for eye graft material that includes galactose, N-
acetylglucosamine (a modified sugar found in the GAGs) and chondroitin sulfate
(a
GAG). Additionally, United States Patent No. 4,486,416 issued to Soll et al.,
discloses a
method of protecting corneal endothelial cells exposed to the trauma of
intraocular lens
implantation surgery by administering a prophylactically effective amount of
chondroitin
sulfate. Also, United States Patent No. 5,141,928 issued to Goldman discloses
the
1 o prevention and treatment of eye injuries using glycosaminoglycan
polysulfates.
United States Patent No. 4,983,580 issued to Gibson, discloses methods for
enhancing the healing of corneal incisions. These methods include the
application of a
corneal motor composition of fibronectin, chondroitin sulfate and collagen to
the incision.
In United States Patent No. 4,801,619 issued to Lindblad, the intraarticular
administration of hyaluronic acid is disclosed for the treatment of
progressive cartilage
degeneration caused by proteoglycan degradation.
The use of a SAM and selenium composition as a nutritional supplement is
disclosed in United States Patent Application Ser. No. 08/725,194 filed by one
of the
present inventors and is herein incorporated by reference. In addition, one of
the
2 o inventors of the present invention has taught, in United States Patent No.
5,587,363 the
combination of an aminosugar, such as glucosamine, and a glycosaminoglycan,
such as
chondroitin, for treatment of degenerative joint diseases. One of the present
inventors has
further taught the optional inclusion of manganese in a composition of an
aminosugar and
a glycosaminoglycan in United States Patent No. 5,364,845.
2 5 Accordingly, it can be seen that there remains a need for compositions
which
- 12 -
CA 02333752 2000-11-30
WO 99/62524 PC'T/US98/11748
include analgesic, anti-inflammatory, and antidepressant components, as well
as
components that provide the building blocks for the production of connective
tissue in
humans and that also protect against the degradation of that tissue.
SUMMARY OF THE INVENTION
It is therefore an object of the present invention to provide a composition
for the
protection and repair and for reducing the inflammation of connective tissue
in humans
and animals.
It is a further object of the present invention to provide compositions which
l0 contain S-Adenosylmethionine and an aminosugar or salts thereof, such as
glucosamine,
for facilitating the repair and reducing the inflammation of connective tissue
in humans
and animals.
It is another object of the present invention to provide compositions which
contain
S-Adenosylmethionine and GAGs, such as chondroitin salts and fragments
thereof, for
facilitating the repair and for reducing the inflammation of connective tissue
in humans
and animals.
It is yet a further object of the present invention to provide compositions
which
contain S-Adenosylmethionine, an aminosugar or salts thereof, and GAGS or
fragments
thereof for facilitating the repair and for reducing the inflammation of
connective tissue in
2 0 humans and animals.
It is another object to optionally provide manganese to any of these
compositions
for humans and animals.
It is still a further object to optionally provide methyl donors or methyl
donor
cofactors, such as vitamins B 12 and B6, folic acid, dimethylglycine, and
trimethylglycine,
2 5 to the compositions of the present invention for humans and animals if
desirable.
- 13 -
CA 02333752 2000-11-30
WO 99/62524 PGT/US9$/11748
It is a further object of the present invention to provide methods of
administering
these compositions.
These and other objects of the present invention will become readily apparent
from a reading of the following detailed description and examples.
BRIEF DESCRIPTION OF THE DRAWINGS
FIG. 1 is a sequence for the biosynthesis of hexosamines.
FIG. 2 is a schematic flowchart illustrating the biological pathway by which
the
composition of the present invention aids in protection and repair of
connective tissue.
FIG. 3 is an enlarged portion of the flowchart of FIG. 2.
DETAILED DESCRIPTION OF THE INVENTION
According to the present invention, a composition selected from the group
consisting of SAM and an aminosugar or salts thereof (e.g., glucosamine); SAM
and
GAGS (e.g., chondroitin salts) or fragments thereof; and SAM, an aminosugar
(or salts
thereof), and GAGs (or fragments thereof) is provided to humans and animals
for
stimulating both collagen and PG synthesis and for reducing inflammation.
Manganese,
preferably manganese salts, may optionally be included to any of these
compositions. In
addition, other optional ingredients include methyl donors or methyl donor
cofactors,
2 o such as vitamins B 12 and B6, folic acid, dimethylglycine, and
trimethylglycine. These
compositions may act to accomplish several functions, including bypassing the
glucose to
glucosamine rate-limiting step in the natural production of proteoglycans in
humans and
animals, and producing additional quantities of collagen and proteoglycans for
use in the
repair of damaged connective tissue. In addition, inflammation of connective
tissue may
2 5 be reduced by the compositions of the invention. The compositions of the
present
- 14 -
CA 02333752 2000-11-30
' WO 99/62524 PCTNS98/11748
invention may achieve these functions directly or through indirect pathways --
i.e.,
through their effect on other components in the living system which in tum can
increase
connective tissue synthesis or reduce inflammation.
In one embodiment, a composition of the present invention include S-
Adenosyimethionine (SAM) and an aminosugar, such as glucosamine, preferably in
a salt
form. In another embodiment of the present invention, the composition includes
SAM
and a glycosaminoglycan, such as chondroitin (preferably in a salt form such
as
chondroitin sulfate). In another embodiment, the composition of the present
invention
includes SAM, an aminosugar, such as glucosamine, preferably in a salt form,
and a
glycosaminoglycan, such as chondroitin (preferably in a salt form, such as
chondroitin
sulfate). Alternatively, fragments of a glycosaminoglycan may be used in a
composition
of the invention in addition to or in substitution for the glycosaminoglycan.
Each of these
compositions may optionally include manganese. A preferred form of manganese
in such
compositions is a manganese salt, such as manganese ascorbate, because the
ascorbate is a
soluble form of manganese which further provides ascorbic acid, a substance
needed for
collagen synthesis. Other manganese salts such, as for example, sulfate or
gluconate, may
be used however. Each of these compositions may optionally contain one or more
methyl
donors or methyl donor cofactors selected from the group consisting of
vitamins B 12 and
B6, folic acid, dimethylglycine, and trimethylglycine.
2 0 Referring to FIGS. 2 and 3, the biosynthetic pathway for the production of
connective tissue, which is affected by the method of the present invention by
virtue of
the components of the composition of the present invention which aid in
connective tissue
repair, functions as described in the above background section of this
application.
In a preferred embodiment, the aminosugar glucosamine is the base of the
2 5 composition, providing the primary substrate for both collagen and
proteoglycan
- 15 -
CA 02333752 2000-11-30
' WO 99/62524 PC'T/US98/11748
synthesis. Glucosamine is the preferred substrate for proteoglycan synthesis,
including
chondroitin sulfates and hyaluronic acid. The glucosamine preferably is in a
salt form so
as to facilitate its delivery and uptake by humans and animals. The preferred
salt forms
are glucosamine hydrochloride, glucosamine sulfate and N-acetylglucosamine.
Administration of a preferred embodiment of the composition of the present
invention provides the human or animal organism with exogenous quantities of
SAM, an
aminosugar or salts thereof, and a glycosaminoglycan or fragments thereof. If
desired,
the composition also provides the human or animal organism with exogenous
quantities
of manganese cofactors. Also if desired, the compositions of the present
invention may
include methyl donors or methyl donor cofactors, such as vitamins B 12 and B6,
folic
acid, dimethylglycine, and trimethylglycine.
The exogenous glucosamine provided by the composition of present invention is
converted to proteoglycans as is seen in FIG. 2 and as described above.
In the former case, the glucosamine may be converted with the aid of manganese
directly into GAG, including hyaluronic acid (which is 50% glucosamine and
which
forms the backbone of the proteoglycans). This core protein is then linked to
the
hyaluronic acid via the link protein, as is seen in FIG. 3.
In the latter case, the free amino acids are, with the aid of manganese and
zinc
cofactors (and ascorbic acid or vitamin C), converted to procollagen. The
procollagen is
2 0 then converted into collagen with the aid of copper or iron cofactors and
vitamin C
(ascorbic acid) and sulfate chelates.
Thus, preferred compositions of the present invention containing SAM and
glucosamine advantageously stimulate the synthesis of collagen and
glycosaminoglycans
or mucopolysaccharides (GAGs), including hyaluronic acid, the backbone of
2 5 proteoglycans (PG's), thereby providing a natural tissue repair function.
These
- 16 -
CA 02333752 2000-11-30
WO 99/G2524 PCT/US98111748
compositions provide the connective tissue repair function of glucosamine, the
increased
sulfation of GAGs by SAM, the stabilization by SAM metabolites of the
polyanionic
macromolecules of proteoglycans, and the additional analgesic, anti-
inflammatory, and
anti-depressant effects of SAM. The optional addition of manganese provides a
further
benefit if a deficiency of the mineral exists or if it is otherwise desired.
The optional
inclusion of methyl donors or methyl donor cofactors, such as vitamins B 12
and B6, folic
acid, dimethylglycine, and trimethylglycine, helps to promote methylation and
thereby
convert homocysteine to methionine.
Another preferred composition of the invention comprises SAM and chondroitin
salts (such as chondroitin sulfate). SAM operates in this composition, in
conjunction with
endogenous glucosamine, as described above. Chondroitin salts operate with SAM
and
endogenous glucosamine by inhibiting the synovial degradative enzymes.
Chondroitin
salts (such as chondroitin sulfate) also directly contribute to the pool of
GAGS of
cartilaginous tissue. Manganese salts may also be included in this composition
in those
cases where a deficiency of manganese exists. Methyl donors or methyl donor
cofactors,
such as vitamins B 12 and B6, folic acid, dimethylglycine, and
trimethylglycine, may
optionally be included in these compositions to help promote methylation and
thereby
convert homocysteine to methionine.
Another preferred embodiment of the composition of the present invention
2 0 contains SAM, glucosamine, and chondroitin salts (such as chondroitin
sulfate) and
mixtures and fragments thereof, and also advantageously stimulates the
synthesis of
collagen and glycosaminoglycans or mucopolysaccharides (GAGS), including
hyaluronic
acid, thereby providing a natural tissue repair function. This composition
provides the
superior connective tissue repair function of glucosamine, the above-described
benefits of
2 5 SAM, and the above-described benefits from chondroitin salts (including
chondroitin
- 17 -
CA 02333752 2000-11-30
WO 99/62524 PCT/US98/11948
sulfate) and fragments of chondroitin salts. Chondroitin salts (including
chondroitin
sulfate) also operate with SAM and glucosamine by inhibiting the synovial
degradative
enzymes. Chondroitin salts (including chondroitin sulfate) also directly
contribute to the
pool of GAGs of cartilaginous tissue. Manganese provides a further benefit if
a
deficiency of the mineral exists. As with the compositions described above,
methyl
donors or methyl donor cofactors, such as vitamins B 12 and B6, folic acid,
dimethylglycine, and trimethylglycine, may optionally be included in these
compositions
to help promote methylation and thereby convert homocysteine to methionine.
Tissue
repair can thus be accomplished, in the context of the treatment and repair of
connective
1 o tissue and the treatment of arthritic conditions, in almost all areas of
the body both human
and animal.
In the present method for the treatment and repair and for reducing the
inflammation of connective tissue in humans and animals, preferred
compositions
comprising amounts of SAM in combination with glucosamine including salts
thereof in
combination with chondroitin salts (including chondroitin sulfate) or
fragments thereof, or
amounts of SAM and chondroitin salts (including chondroitin sulfate) or
fragments
thereof in combination with glucosamine including salts thereof, may be
administered to
humans and animals thereof for stimulating both collagen and proteoglycan
synthesis. An
additional preferred composition comprising amounts of SAM and chondroitin
salts
2 0 ~ {including chondroitin sulfate) or fragments thereof may be administered
to humans and
animals for stimulating proteoglycan synthesis and reducing inflammation.
Manganese
salts may also be optionally included in each composition in cases where a
deficiency of
manganese exists. Methyl donors or methyl donor cofactors, such as vitamins B
12 and
B6, folic acid, dimethylglycine, and trimethylglycine may optionally be
included to these
2 5 compositions as well.
- 18 -
CA 02333752 2000-11-30
' WO 99/62524 PCT/US98/11748
The compositions of the present invention are administered to promote tissue
repair, including cartilage repair, and the treatment of arthritic conditions
as well as
connective tissue damage in humans and animals. The anti-depressant effect of
SAM
may help to alleviate the burden of sickness for some patients, thus enhancing
their
quality of life. This effect, as well as the analgesic and anti-inflammatory
effects of SAM
which will help alleviate the pain associated with arthritic conditions, may
help remove
impediments to physical activity. Increased levels of physical activity, in
turn, can supply
the loading and unloading forces necessary for the regeneration of articular
cartilage.
Supplementation with glucosamine, with its chondroprotective role, thus helps
to ensure
that the raw materials are available to support the increased regeneration of
cartilage. The
compositions of the present invention are also understood to play a role in
chondromodulation, chondrostabilization, and chondrometabolizaton.
The dosage of SAM in the nutritional supplements of the present invention
ranges
from about S mg to about 5,000 mg in humans and small animals, and from about
2 mg to
about 20,000 mg in large animals (e.g., equine). The dosage of giucosamine in
the
nutritional supplements of the present invention ranges from about SO mg to
about 5,000
mg in humans and small animals, and from about 2S0 mg to about 40,000 mg in
large
animals (e.g., equine). The dosage of chondroitin salts in the nutritional
supplements of
the present invention ranges from about 1 S mg to about 5,000 mg in humans and
small
2 0 animals, and from about 100 mg to about 30,000 mg in large animals. When
included in
the compositions of the present invention, manganese may optionally be present
in the
range of about 2 to about 7S mg in humans and small animals, and from about 10
mg to
about S00 mg in large animals. The ascorbate component of the manganese
ascorbate
may range from about 10 mg to about S00 mg in humans and small animals, and
from
2 5 about SO mg to about 2,500 mg in large animals. When included in the
compositions of
- 19 -
CA 02333752 2000-11-30
' WO 99162524 PCT/US98/11748
the present invention, the methyl donors or methyl donor cofactors, such as
vitamins B 12
and B6, folic acid, dimethylgiycine, and trimethylglycine may be present in
the range of
about 0.1 mg to about 10 mg in humans and small animals, and from about 1 mg
to about
100 mg in large animals.
As a preferred embodiment, a dosage of the nutritional supplement composition
of
the present invention may consist of one or more capsules or tablets for human
oral
consumption. In such an embodiment, the preferred weight of the dosage is
between
about 5 mg to about 5,000 mg, and preferably about 2,500 mg. The dosage may be
administered in a single daily dosage form in which all components are
present, e.g., a
capsule or tablet of preferably 2,500 mg. The dosage may also be administered
in more
than one dosage form in which each dosage form contains at least one
component. When
a single dosage is administered in more than one dosage form, the multiple
dosage forms
may be co-administered as a single dosage. Thus, for example, a single dosage
may be
comprised of a SAM dosage form co-administered with a glucosamine and
chondroitin
salts dosage form.
Alternatively, the nutritional supplement compositions of the present
invention
may be administered more than once daily. Hence, for example, the nutritional
supplement compositions of the present invention may be in the form of an oral
dosage
form of 1250 mg administered twice daily or 833 mg administered three times
daily. The
2 o number of daily administrations will depend upon the needs of the human or
animal
recipient. Different connective tissue disorders and injuries require
different amounts of
the compositions of the present invention. In that regard, several dosages may
be
administered depending on the particular needs of the human or animal.
Alternatively, and of particular use in large animals, the compositions of the
2 5 present invention may for example be administered in scoops. Such
administration may
- 20 -
CA 02333752 2000-11-30
WO 99/62524 PCT/US98/11748
take the form, for example, of a level scoopful containing about 1,800 mg
glucosamine,
about 600 mg chondroitin salts, about 16 mg of manganese (when included in the
form of
manganese ascorbate), and about 104 mg of ascorbate (when included in the form
of
manganese ascorbate).
These preparations may be made by conventional methods. For example, to
prepare the compositions of the invention, the above-described ingredients are
combined
as the active ingredient in intimate admixture with a suitable carrier
according to
conventional compounding techniques. This Garner may take a wide variety of
forms
depending upon the form of preparation desired for administration, e.g., oral,
sublingual,
nasal, guttural, rectal, transdermal or parenteral.
In preparing the compositions in oral dosage form, any usual pharmaceutical
medium may be employed. For oral liquid preparations (e.g., suspensions,
elixirs, and
solutions), media containing for example, water, oils, alcohols, flavoring
agents,
preservatives, coloring agents and the like may be used. Carriers such as
starches, sugars,
diluents, granulating agents, lubricants, binders, disintegrating agents, and
the like may be
used to prepare oral solids (e.g., powders, capsules, pills, caplets, tablets,
microencapsulated granules, microtablets, coated granules and lozenges).
Capsules or
tablets are a preferred oral dosage form. Controlled release forms may also be
used.
Because of their ease in administration, lozenges, tablets, pills, caplets,
and capsules
2 0 represent the most advantageous oral dosage unit form, in which case solid
pharmaceutical carriers are obviously employed. If desired, tablets may be
sugar coated
or enteric coated by standard techniques. The compositions of the present
invention may
be in the form of one or more of these oral dosage forms -- i.e., a single
dosage may be in
multiple forms.
- 21 -
CA 02333752 2000-11-30
' WO 99/62524 PGT/US98111748
For parenteral products, the carrier will usually comprise sterile water,
although
other ingredients may be included, e.g., to aid solubility or for preservation
purposes.
Injectable suspensions may also be prepared, in which case appropriate liquid
carriers,
suspending agents, and the like may be employed.
Having discussed the composition of the present invention, it will be more
clearly
perceived and better understood from the following specific examples which are
intended
to provide examples of the preferred embodiments and do not limit the present
invention.
EXAMPLE 1
1 o The composition of the present invention is made in one or more capsules
for oral
administration in humans and small animals. In a preferred embodiment, each
dosage
contains:
Human & Small Animal Ran"g_e/Dose
SAM 5-5,000 mg
Glucosamine 50-5,000 mg
Chondroitin Sulfate 15-5,000 mg
2 o EXAMPLE 2
For those situations in which
manganese supplementation is
desired, a manganese
salt is added to the composition 1 so that each dosage contains:
of Example
Human & Small Animal Ran eg /Dose
2 5 SAM 5-5,000 mg
Glucosamine 50-5,000 mg
Chondroitin Sulfate 15-5,000 mg
Manganese (as Ascorbate) 2-75 mg
Ascorbate (as Manganese
3 o Ascorbate) 10-500 mg
- 22 -
CA 02333752 2000-11-30
WO 99/62524 PCT/US98/11748
EXAMPLE 3
For larger animals, such as horses, the composition of Example 1 is
administered
as filled scoops.
Lar~,e Animal~Eauin~ Range/Dose
S~ 2-20,000 mg
Glucosamine 250-40,000 mg
Chondroitin Sulfate 100-30,000 mg
EXAMPLE 4
For those situations in which
manganese supplementation is
desired, manganese
salts may be added to the composition
of Example 3 so that each dosage
contains:
Lame Animal (Equine) Range/Dose
SAM 2-20,000 mg
Glucosamine 250-40,000 mg
Chondroitin Sulfate 100-30,000 mg
Manganese (as Ascorbate) 10-500 mg
Ascorbate (as Manganese
2 0 Ascorbate) 50-2,500 mg
EXAMPLE 5
For a further preferred composition, each dosage contains:
Human & Small Animal Range/Dose
SAM 5-5,000 mg
Glucosamine 50-5,000 mg
3 o EXAMPLE 6
For those situations in which manganese supplementation is desired, a
manganese
salt is added to the composition of Example 5 so that each dosage contains:
Human & Small Animal Ran eg_/Dose
- 23 -
CA 02333752 2000-11-30
WO 99/62524 PCTNS98/11748
S~ 5-5,000 mg
Glucosamine 50-5,000 mg
Manganese (as Ascorbate) 2-75 mg
Ascorbate (as Manganese
Ascorbate) 10-500 mg
EXAMPLE 7
For larger animals, such as horses, the composition of Example 5 is
administered
as filled scoops.
1 o Lar,~e Animal (Equinel Ranae/Dose
SAM 2-20,000 mg
Glucosamine 250-40,000 mg
EXAMPLE 8
For those situations in which
manganese supplementation is
desired, manganese
salts may be added to the composition
of Example 7 so that each dosage
contains:
Lame Animal (Equine, Range~ose
2 o SAM 2-20,000 mg
Glucosamine 250-40,000 mg
Manganese (as Ascorbate) 10-S00 mg
Ascorbate (as Manganese
Ascorbate) 50-2,500 mg
EXAMPLE 9
For a further preferred composition, each dosage contains:
Human & Small Animal Ran e/Dose
3 0 SAM 5-5,000 mg
Chondroitin Sulfate 15-5,000 mg
- 24 -
CA 02333752 2000-11-30
WO 99/62524 PCT/US98/11748
EXAMPLE 10
For those situations in which manganese supplementation is desired, a
manganese
salt is added to the composition of Example 9 so that each dosage contains:
Human & Small Animal Range/Dose
SAM S-5,000 mg
Chondroitin Sulfate 1 S-5,000 mg
Manganese (as Ascorbate) 2-7S mg
Ascorbate (as Manganese
Ascorbate) 10-S00 mg
EXAMPLE 11
For larger animals, such as horses, the composition of Example I O is
administered
as filled scoops.
Lame Animal (Equines Ran~e/Dose
SAM 2-20,000 mg
Chondroitin Sulfate 100-30,000 mg
EXAMPLE 12
For those situations in which manganese supplementation is desired, manganese
salts may be added to the composition of Example 11 so that each dosage
contains:
2 5 Lame Animal (Eguine) Ran;~e/Dose
SAM 2-20,000 mg
Chondroitin Sulfate 100-30,000 mg
Manganese (as Ascorbate) 10-S00 mg
Ascorbate (as Manganese
3 0 Ascorbate) SO-2,500 mg
EXAMPLE 13
For those situations in which methyl donors or methyl donor cofactors are
desired,
- 25 -
CA 02333752 2000-11-30
WO 99/62524 PCT/US98/11748
such compounds may be added to the composition of Example 1 so that each
dosage
contains:
Human & Small Animal Ran eg /Dose
SAM 5-5,000 mg
Glucosamine 50-5,000 mg
Chondroitin Sulfate 15-5,000 mg
vitamin B 12 0.1-10 mg
EXAMPLE 14
For those situations in which
manganese supplementation is
desired, a manganese
salt is added to the composition 13 so that each dosage contains:
of Example
Human & Small Animal Ranae/Dose
SAM 5-5,000 mg
Glucosamine 50-5,000 mg
Chondroitin Sulfate 15-5,000 mg
Manganese (as Ascorbate) 2-75 mg
2 0 Ascorbate (as Manganese
Ascorbate) 10-500 mg
vitamin B12 0.1-10 mg
EXAMPLE 15
2 5 For larger animals, such as horses, the composition of Example 13 is
administered
as filled scoops.
Larne Animal (Equine) Ran. e/Dose
SAM 2-20,000 mg
3 0 Glucosamine 250-40,000 mg
Chondroitin Sulfate 100-30,000 mg
vitamin B12 1-100 mg
EXAMPLE 16
3 5 For those situations in which manganese supplementation is desired,
manganese
salts may be added to the composition of Example 15 so that each dosage
contains:
- zs -
CA 02333752 2000-11-30
WO 99/62524 PCT/US98/11748
Lame Animal (Equine) Ran; e/Dose
S~ 2-20,000 mg
Glucosamine 250-40,000 mg
Chondroitin Sulfate 100-30,000 mg
Manganese (as Ascorbate) 10-500 mg
Ascorbate (as Manganese
Ascorbate) 50-2,500 mg
vitamin B12 1-100 mg
1 o EXAMPLE 17
For a further preferred composition, each dosage contains:
Human & Small Animal Ran e,~ /Dose
SAM 5-5,000 mg
Glucosamine 50-5,000 mg
vitamin B12 0.1-10 mg
EXAMPLE 18
For those situations in which
manganese supplementation is
desired, a manganese
2 0 salt is added to the composition17 so that each dosage contains:
of Example
Human & Small Animal Ranae/Dose
SAM S-5,000 mg
Glucosamine 50-5,000 mg
2 5 Manganese (as Ascorbate) 2-75 mg
Ascorbate (as Manganese
Ascorbate) 10-500 mg
vitamin B12 0.1-10 mg
3 0 EXAMPLE 19
For larger animals, such as horses, the composition of Example 17 is
administered
as filled scoops.
La~.~ee Animal (Equine) Range/Dose
3 5 SAM 2-20,000 mg
Glucosamine 250-40,000 mg
vitamin B 12 1-100 mg
- 27 -
CA 02333752 2000-11-30
WO 99/62524 PC'T/US98/11748
EXAMPLE 20
For those situations in which manganese supplementation is desired, manganese
salts may be added to the composition of Example 19 so that each dosage
contains:
Laree Animal (Equine)
SAM 2-20,000 mg
Glucosamine 250-40,000 mg
Manganese (as Ascorbate) 10-500 mg
Ascorbate (as Manganese
1 o Ascorbate) 50-2,500 mg
vitamin B 12 1-100 mg
EXAMPLE 21
For a further preferred composition, each dosage contains:
Human & Small Animal Ran~e/Dose
S~ 5-5,000 mg
Chondroitin Sulfate 15-5,000 mg
vitamin B 12 0.1-10 mg
EXAMPLE 22
For those situations in which
manganese supplementation is
desired, a manganese
salt is added to the composition21 so that each dosage contains:
of Example
Human & Small Animal Range/Dose
SAM S-5,000 mg
Chondroitin Sulfate 15-5,000 mg
Manganese (as Ascorbate) 2-75 mg
Ascorbate (as Manganese
3 0 Ascorbate) 10-500 mg
vitamin B 12 0.1-10 mg
- 28 -
CA 02333752 2000-11-30
WO 99/62524 PCT/US98/11'748
EXAMPLE 23
For larger animals, such as horses, the composition of Example 21 is
administered
as filled scoops.
Lame Animal (Equine) Range/Dose
SAM 2-20,000 mg
Chondroitin Sulfate 100-30,000 mg
vitamin B 12 1-100 mg
1 o EXAMPLE 24
For those situations in which manganese supplementation is desired, manganese
salts may be added to the composition of Example 23 so that each dosage
contains:
Lame Animal (E4uine~ Rarn~ e/Dose
S~ 2-20,000 mg
Chondroitin Sulfate 100-30,000 mg
Manganese (as Ascorbate) 10-500 mg
Ascorbate (as Manganese
Ascorbate) 50-2,500 mg
vitamin B 12 1-100 mg
Many modifications may be made without departing from the basic spirit of the
present invention. Accordingly, it will be appreciated by those skilled in the
art that
within the scope of the appended claims, the invention may be practiced other
than has
been specifically described herein.
- 29 -
