Note : Les descriptions sont présentées dans la langue officielle dans laquelle elles ont été soumises.
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~ITLE OF INVENTION
FORMULATIONS CONTAI~ING HYALURONIC ACID
FIELD OF INVENTIO~
This invention relates to the treatment of disease and
conditions of the skin and exposed tissue. In some embodiments
this invention finds application to the treatment of a disease
or condition of the skin and exposed tissue including basal cell
carcinoma, squamous cell tumours, metastatic cancer of the
breast to the skin, primary and metastatic melanoma in the skin,
malignancies and tumours in the skin, genital warts (condyloma
acuminata), cervical cancer, HPV (Human Papilloma Virus)
including HPV (Human Papilloma Virus) on the cervix, psoriasis
(both plaque-type psoriasis and nail bed psoriasis), corns on
the feet, actinic keratoses lesions, "liver" spots, fungal
lesions, and other such types of lesions, and hair loss on the
head of a pregnant women.
This invention also relates to compositions and
formulations suitable for use in such treatments, the use of
such formulations in such treatments, methods of such treatment,
and the delivery of drugs for such treatments.
~B~CKGROUND OF THE INVENTION
Basal cell carclnoma is presently treated by surgery.
Each lesion, together with all surrounding and underlying tissue
(dermis, epidermis, and subdermis), is cut out. In some
instances, surgery, while necessary for the patient's welfare,
puts the patient at risk in some other respect (for example, the
removal of a lesion on a patient's temple (resection) may
jeopardize the patient's health). Squamous cell tumours are also
treated the same way as are other forms of cancer in the skin
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and exposed tissue. Furthermore, other conditions and diseases
of the skin and exposed tissue are treated in the same way or in
ways that cause discomfort to the patient, for example melanoma,
genital warts, cervical cancer, HPV (Human Papilloma Virus).
Actinic keratoses lesion is dealt with similarly.
Liquid nitrogen is also used to remove the lesion.
These dlseases and conditions are usually found in the
epidermis (at least for the most part, extending into the dermis
and upwards through the Stratum Corneum).
Hyaluronic acid is a naturally occurring
glycosaminoglycan. Its molecular weight may vary from 50,000
daltons upwards, and it forms highly viscous solutions. As
regards the actual molecular weight of hyaluronic acid in
natural biological contexts, this is still a matter of much
uncertainty; when the molecular weight of hyaluronic acid is to
be determined, different values are obtained depending on the
assay method employed, and on the source, the isolation method
etc. The acid occurs in animal tissue, e.g. spinal fluid, ocular
fluid, synovial fluid, cockscombs, skin, and also in some
streptococci. Various grades of hyaluronic acid have been
obtained. A preparation with an allegedly high degree of purity
and alleged to be entirely free from side effects, is a non-
inflammatory form described in U.S. Patent No.4,141,973; this
preparation i9 said to have a molecular weight exceeding~750,000
dalton, preferably exceeding 1,200,000 dalton and is suggested
for therapeutic use in various articular conditions. Applicants
believe that hyaluronic acid claimed in this patent is sold
under the trade mark Healon.
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United States Patent 4,801,619 relates to hyaluronic
acid,having a molecular weight of about 3 X 10~ dalton or more,
administered intra-articularly which is prone to decrease the
proteoglycan content of synovial fluid to almost normal levels.
According to this patent, this indicates a positive effect on
the proteoglycan metabolism of a joint. According to the
patent, this is applicable both to inflammatory conditions and
to degeneration caused by treatment with symptomatics, such as
corticosteroid preparations. It is thus clear that a
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sufficiently high molecular weight of the hyaluronic acid is
alleged to counteract side effects that might be caused by
corticosteroids or other symptomatics produclng similar effects.
When corticosteroids are applied, the amount of hyaluronic acid
in the synovial cavity will, according to the patent, increase
substantially and, according to the inventors, their hyaluronic
acid preparations have a very positive effect on such clinical
symptoms as pain, swelling, and lameness.
The patent states that the objectives of the invention
are attained by intra-articular administration (injection) of an
effective amount of hyaluronic acid with a mean molecular weight
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exceeding 3 X 106 dalton, preferably exceeding 4 X 106 dalton;
usually the molecular weight will not exceed 7 X 106 dalton.
The dosage of hyaluronic acid administered is stated to be ;~
preferably within the range of Smg-80mg. The amount of solution
25 given at each administration is generally less than 60 ml, e.g. ~;
less than 20 ml. of an aqueous solution of the acid or its salt. ~
It is convenient to administer the acid dissolved in water (<2% -
w/w, buffered to physiological pH), for instance in the form of
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a water-soluble sodium salt. The exact amount will depend on
the particular joint to be treated.
The Merck Index Specifies that Hyaluronic Acid has a
Molecular Weight within the range pf 50,000 to 8 X 106
depen ding on source, methods of preparation, and methods of
determination. The Merck Publication teaches hyaluronic acid as
a surgical aid (ophthalmological).
United States Patent 4,808,576 purports to teach that
hyaluronic acid, an agent well known for reducing the sequelae
of trauma in mammalian joint tissue when injected directly into
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the traumatized joint tissue, will be carried to such
traumatized tissue by the mammal's natural processes if applied
at a site remote from the traumatized tissue. Thus, hyaluronic
acid in any therapeutically acceptable form can, according to
the Patent, be administered by the typical remote routes
including intravenous, intramuscular, subcutaneous, and topical.
This, the patent alleges, makes the utilization of
hyaluronic acid much more convenient and attractive. For
instance, the treatment of arthritis in horse or human joints
with hyaluronic acid, according to the patent, no longer
requires more difficult intra-articular injections.
United States Patent 4,725,585 relates to a method of
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enhancing or regulating the host defence of a mammal by
administering to a mammal a therapeutically effective amount of
hyaluronic acid.
At column 1, lines 43 - 46, the patent provides that :
the invention was based on the unexpected discovery that
administration of hyaluronic acid to mammals results in a
considerable increase in the defence.
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The hyaluronic acid employed in the patent was Healon
(t.m) provided by Pharmacia AB, Uppsala, Sweden (Pharmacia AB is
also entitled to the benefit of United States Patent 4,141,973).
The patent provides at column 4, line 19 that because a
patient's infections had been hard to treat, instead of just
hyaluronic acid being administered to the patient to increase
the patient's defence, the patient was given hyaluronic acid and
an antibiotic. While one reading the patent may conclude that
the antibiotic was given in combination with hyaluronic acid, in
fact because the hyaluronic acid was administered subcutaneously
and because the patient was a heart patient, one skilled in the
art would understand that any antibiotic administered, while
possibly administered simultaneously with the administration of
the hyaluronic acid, was definitely administered separately
intravenously (probably) or intramuscularly (less probably).
Thus, the hyaluronic acid administered, according to the
teachings of this patent, was administered in order to prevent
possible development of infections (increase the host's defence)
and not for any other reason.
~nited States Patent 4,636,524 discloses cross-linked
gels of hyaluronic acid, alone and mixed with other hydrophilic
polymers and containing various substances or covalently bonded
low molecular weight substances and processes for preparing
them. These products are alleged to be useful in numerous
applications including cosmetic formulations and as drug
delivery systems.
The patent further states that as hyaluronic acid is
known to be a biologically tolerable polymer in the sense that
it does not cause any immune or other kind of response when
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introduced into a human body, the cross-linked hyaluronic acid
gels can be used for various medical applications. The cross-
linked gels modified with other polymers or low molecular weight ~ ~-
substances, it is alleged, can be used as drug delivery devices.
For example, the inventors are alleged to have found that
heparin introduced in a cross-linked hyaluronic acid gel
retained its antithrombogenic activity.
The inventors also allege that they have also found
that cross-linked gels of hyaluronic acid can slow down the
release of a low molecular weight substance dispersed therein
but not covalently attached to the gel macromolecular matrix. :
Unites States Patent 4,736,024 purports to teach new
medicaments for topical use containing:
(i) an active pharmacological substance or a mixture
lSof pharmacological substances, either active or
suitable for topical administration and
(ii) a topical vehicle which comprises hyaluronic
acid or a molecular fraction of hyaluronic acid or a salt of the
same with an alkaline metal, an alkaline earth metal, magnesium,
aluminium, ammonium, or a pharmacological substance optionally
together with additional conventional excipients for ;
pharmaceutical preparations for topical use.
Applicants are also aware of published Japanese Patent -
Document 61000017, dated 86/01/06, whose English abstract of
disclosure states that the Japanese Patent Document relates to
the use of hyaluronic acid or cross-linked hyaluronic acid or
their salts as the active ingredient fcr inhibiting carcinoma
metastasis. ;
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According to the purported abstract of the patent,
more that 1.0% of hyaluronic acid is dissolved in alkaline aq.
soln. and pref. more than 50~ of H20 sol. org. solvent. eq.
alcohol, acetone, dioxane, against total soln. is added.
Preferably the pH is 12-14. Then a multifunctional epoxy cpd.
is added and reacted at 10-60 deg. C, pref. at 20-40- deg. C for
24 hrs. Cross-linking ratio of crosslinked hyaluronic acid or
its salt is regulated by changing mol ratio of hyaluronic acid
or its salt and multifunctional epoxy cpd.. Pref. hyaluronic
acid used has intrinsic viscosity 0.2-30,m.w. 4000-2000000. The
hyaluronic acid is allegedly used in several dosage forms. The
clinical dose for an adult is alleged to be normally, as
hyaluronic acid or cross-linked hyaluronic acid, 25mg-5 g/day
~p.o.) and 10 mg-2.5 g/l dose (inj). The abstract alleges that
the advantage is that the hyaluronic acid has no side effects as
may other anti-cancer drugs and has an analgesic and a tissue
restoration effect.
European Patent Application 0295092 purports to teach
a vehicle together with fragments of hyaluronic acid for
delivering of the fragments of hyaluronic acid into the skin to
reach the dermal layer of the skin to increase the development
of blood vessels for stimulating hair growth or regrowth. The
preferred fragments of hyaluronic acid are polysaccharides
containing from 7 to 25 monosaccharide units. The patent
provides that it is apparent that the larger the fragments of
hyaluronic acid, the greater the difficulty there is in
delivering the fragments to the dermal layer o-f the skin, unless
there is also present in the composition a means for enhancing
the activity of said fragments.
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The combination may thus include a means for enhancing
the activity of the fragments of hyaluronic acid, especially to
improve their penetration through the skin following topical
application. Some activity enhancers, it is alleged, also
function as vehicles for the fragments of the hyaluronic acid.
Some activity enhancers are also alleged to possess
the ability to stimulate or increase hair growth. Minoxidil is
asserted among others to be such an activity enhancer. Thus
both the fragments of hyaluronic acid and minoxidil are alleged
to stimulate hair growth both delivered by a vehicle.
European Patent Application 0179442 asserts that where
free radicals are formed in considerable quantities, hyaluronic
acid is broken down or degraded before the hyaluronic acid has
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given the desired effect.
Canadian Letters Patent 1,240,929 teaches the :
combination of chondroitin sulfate compound and a hyaluronate to
protect both human and animal cell layers and tissue subject to
exposure to trauma. ~ .
European Patent Application 0208623 purports to teach
hyaluronic acid as "une augmentation de l'activité de certaines
proteases". It also purports to teach the use of hyaluronic
acid for treating connective tissue diseases, including
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malignant tumours and cardiovascular disorders. -
European Patent Application 270317 purports t`o teach
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the combination of an antiviral agent lacking inhibitory action
and a compound [for example, hyaluronic acid] possessing cell
fusion inhibitory activity and/or virus-adsorption inhibitory ;
activity for treating disease carried by a virus.
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United States Patent 4,840,941 purports to teach the
use of an effective amount of hyaluronic acid as the active
agent for the treatment of retroviruses in association with a
pharmaceutically acceptable carrier, diluent, or excipient.
United States Patent 4,851,521 and European Patent
Application 0265116 both describe hyaluronic acid fractions,
the making thereof and cross-linked esters of hyaluronic.
United States Patent 4,851,521 describes esters of hyaluronic
acid incorporated into pharmaceutical preparations as the active
ingredient and as vehicles for ophthamological medicines for
topical use (See column 11, lines 35 to 42; and column 12, lines
62 to column 13, line 3) and in suppositories for a systemic
effect due to the effect of transcutaneous absorption, such as
in suppositories.
lS The patent provides at column 13, lines 5 to 31:
"The vehicling action of the hyaluronic esters
also applies to associated medicaments of the
type mentioned above in which the active
substance acts not only topically or by nasal or
rectal absorption, for example by nasal sprays or
preparations for inhalation for the oral cavity
or the pharynx, but also by oral or parenteral
route, for example by intramuscular, subcutaneous
or intravenous route, as it favors absorption`of
the drug into the application site. The new
medicaments can therefore be applied, apart from
in the fields already mentioned, in practically
all sectors of medicine, such as internal
medicine, for example in pathologies of the
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cardiovascular system, in infections of the
respiratory system, the dlgestive system, the
renal system, in diseases of an endocrinological
nature, in oncology, in psychiatry etc., and may
also be classified therefore from the point of
view of their specific action, being perhaps
anesthetics, analgesics, anti-inflammatories,
wound healers, antimicrobics, adrenergic agonists
and antagonists, cytostatics, antirheumatics,
antihypertensives, diuretics, se~ual hormones,
immunostimulants and immunosuppressants, for
example, one of the drugs having the activity
already described for the therapeutically active
alcohols to be used as esterifying component
lS according to the present invention, or for the
therapeutically active bases used for the
salification of the free carboxylic groups."
There have been extensive studies to determine the
defect in immune function that allows a tumour cell to develop.
It was postulated initially by Jerne, and subsequently by
Burnett, that the immune system's major role was that of
immunological surveillance to destroy abnormal cells. The
concept of surveillance, while somewhat simplistic, remains an
accepted concept for the elaborate mechanism of immune
recognition and function that is present in the higher species -
mammals.
It has then been postulated that tumours develop
because of local or generalized immune suppression. However, as
pointed out by Moller, if general immune suppression occurs, it
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is only certain types of neoplastic disorders that develop,
mainly those of the lympho-reticular system. This observation
is generally correct and represents a major challenge to the
immune surveillance theory unless a specific reason can be shown
as to why the individual cancer cell can develop plus
individually evade the immune system.
It was demonstrated experimentally in 1974 that
defects of macrophage function may exist in neoplastic disease.
The initial experiments found suppressor cells to be
part of the immune system; these were either of the T-cell type
of the macrophage cell system. There was presence demonstrated
in neoplasia, chronic bacterial infection, recovery from massive
injury and chronic fungal infection.
There has been repeated demonstration in experimental
animals that the macrophage cell function is altered in
neoplastic disease. The macrophages in the animal's systems
appeared "blocked" in their function. Generally when removed
from the in vivo situation, washed in saline and cultured, they
perform normally. This block has been shown to be related to
the excessive production of prostaglandin by neoplastic tissue
or by the macrophage itself. Similarly, the N.K. cells ~which
are said to be primitive or immature macrophages and which may
be involved in cancer defence) are also blocked.
In the basic research efforts in the latter '70s and
the early '80's, there existed considerable confusion as to what
role immunotherapy should take in cancer. Activation or
"hyping" of macrophages was thought to be important. However,
in an examination by Romans and Falk of peritoneal macrophages
obtained from patients with neoplastic disease, there was
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definite evidence that these macrophages were already activated
yet were co-existing with cancer cells and not causing their
destruclion.
It has recently been shown by several independent
investigators that the malfunction of macrophages or the
putitive block is due to excessive prostaglandin and that this
can be altered in tissue culture by corticosteroids, ASA, and
the non-steroidal anti-inflammatory drugs, i.e. indomethacin and
naproxen (NaprosynTM). Again, it was repeatedly demonstrated ~:
that in animal tumours these substances could alter the response
to neoplastic cells and that various combinations of these
substances employed with immune enhancing agents could produce
very credible success in eliminating experimental tumours. Lala
and co-workers combined Indomethacin therapy with Interleukin 2
and showed that this could effect a cure with experiment
neoplasm.
There were continued problems with the use of any of
these agents in the actual human in vivo experience. All of the
non-steroidal anti-inflammatory agents (NSAID) produced major
toxicity in terms of gastro-intestinal, neurological, and other
areas. Thus, the basis of the present approach is that, under
general circumstances, with the use of these agents in human
disease in sufficient amounts, the drug will penetrate to any
`` pathological t~issue to alter therapeutically local prostaglandin
production. While intravenous preparations of Indomethacin (and
now of other agents) exist, using these drugs alone produces
prohibitive side effects in human subjects. Therefore, only
insufficient amounts can be brought into the body to effect more
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than occasional responses in neoplasm. :
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However, the majority of the evidence is present to
indicate and therefore, it can be postulated that the basis for
neoplastic development and how the initial cell "sneaks by" the
immune surveillance mechanism relates to its production of
prostaglandin. One need postulate only one mutation to alter
the amount of prostaglandin synthesis produced by cells when
they become "malignant" to establish a mechanism of blocking out
the initial cell in any immune reaction, i.e. the macrophage.
It therefore became essential to develop a combination of NSAIDs
for clinical use to produce a major improvement in response in
neoplastic disease and other conditions where excessive
prostaglandin synthesis represents the basis of the pathogenesis
of this disease state, i.e. arthrltis and various others of the
so-called connective tissue inflammatory disorders and/or auto-
aggressive diseases.
See also:
1. Modulation of Immunity in Cancer Patients by
Prostaglandin Antagonists, Immunitv to Cancer II, Alan R. Liss,
Inc.; and
2. Goodwin, J.S., (1981) Prostaglandin E and Cancer
Growth Potential for Immunotherapy with Prostaglandin Synthesis
Inhibitors, Auamentive Aaents in Cancer Therapy, Raven Press,
New York.
United States Patent 4,711,780 teaches a
pharmaceutical composition comprising Vitamin C, a zinc salt,
and a sulfur amino acid for trea~ing surface epithelium for
epithelium regeneration. Hyaluronic acid may be added for
applications in the reproductive tract to block the passage of
toxins into the blood system.
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U.S. Patent 4,937,259 (Ethicon) teaches combinations
of hyaluronic acid and salts thereof with NSAIDS for the
prevention of adhesions after surgery.
Because of the side effects of the use of non-
steroidal anti-inflammatory drugs ~major toxicity in terms of
gastro-intestinal, neurological, and other areas),use thereof
should also be restricted (if possible) to the area of use
without delivery to other areas which are not in need of
treatment. Thus, if useful amounts of the non-steroidal anti-
inflammatory drugs or for that matter any drugs could bedelivered to a site in need thereof without carriage of
substantial amounts away from the site to be treated, thereby
accumulating an amount of the drug at the site to be treated for
a prolonged period of time, then the use of the drug for example
a non-steroidal anti-inflammatory drug at a site may have many
other useful applications.
SUMMARY OF THE INVENTION
Applicants have now developed compositions,
~combinations and formulations) which are topically applied to
the skin and/or exposed tissue of a human and which are quickly
transported in dosage amounts percutaneously (intracutaneously)
at a site in need of treatment, (site of pathology and/or
trauma) best targeting the epidermis and subsequently remaining
(accumulating) at the site for a prolonged period of time. The
compositions subsequently clear through the lymphatics thereby
bringing dosage amounts of the compositions to the lymphatics
for the treatment of disease and conditions in the lymphatics.
These compositions, (combinations and formulations)
employ, combine, or incorporate (as the case may be) a plurality
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of effective non-toxic dosage amounts, each dosage amount
comprising an effective non-toxic dosage amount of a drug for
example a drug which inhibits prostaglandin synthesis for
example an NSAID and an effective non-toxic dosage amount of a
form of hyaluronic acid (preferably hyaluronic acid or salt
thereof) for the transport of the drug to the site of the
pathology and/or trauma. Suitable dosage amounts of the
composition may be removed from a container (for example a tube
or jar) and administered (for example, applied).
Thus according to one aspect of the invention these
pharmaceutical compositions (combinations and formulations)
comprise a plurality of effective non-toxic dosage amounts for
administration to the skin and/or exposed tissue of a human in
need of treatment, each such dosage amount comprising a
therapeutically effective non-toxic (to the patient) dosage
amount of a drug to treat a disease and/or condition for example
a drug which inhibits prostaglandin synthesis, preferably being
a non-steroidal anti-inflammatory drug (NSAID), for example,
diclofenac, indomethacin, naproxen, and (+/-) tromethamine salt
of ketorolac (sold under the trademark ToradolTM) and an
effective non-toxic dosage amount (for example in excess of 5 mg
per cm2 (square centimeter) of skin or exposed tissue to which
the dosage amount of the composition is to be applied) of
! I hyaluronic aclid and/or salts thereof (for example the sodium
salt) and/or homologues, analogues, derivatives, complexes,
esters, fragments, and/or sub units of hyaluronic acid
~preferably hyaluronic acid and/or salts thereof) to transport
(to faciliate or cause the transport of) the drug to the site of
the pathology and/or trauma of the disease or condition. These
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compositions may be applied topically to treat diseases and
conditions of the skin and/or exposed tissue at the site of the
trauma and/or pathology, (for example, basal cell carcinoma, the
precancerous, often recurrent, actinic keratoses lesions, fungal
lesions, "liver" spots and like lesions (found for the most part
in the epidermis), squamous cell tumours, metastatic cancer of
the breast to the skin, primary and metastatic melanoma in the
skin, malignancies and/or tumours in the skin, genital warts
(condyloma acuminata), cervical cancer, and HPV (Human Papilloma
Virus) including HPV of the cervix, psoriasis (both plaque type
psoriasis and nail bed psoriasis), corns on the feet and hair
loss on the head of pregnant women). The results of the
treatment with suitable dosage amounts taken from these
compositions (combinations and formulations) have been in some
lS instances quite dramatic - difficult situations have been
successfully treated and resolved.
Furthermore, application of the dosage amounts of the
compositions, combinations and formulations are, systemic
independent (there is a lack of a blood level of the drug for
example NSAID), are quick to penetrate into the skin to the site
of the trauma and/or pathology because the effective dosage
amount of the form of hyaluronic acid transports (facilitates or
causes the transport of) the drug (for example NSAID)
! partlicularly to the epidermis where the composition, combination
or formulation accumulates and remains for prolonged periods.
The compositions subsequently clear through the lymphatics and
are available for the treatment of disease and conditions of the
lymphatics.
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In this regard effective amounts of the form of
hyaluronic acid exceeds in the order of about 5 mg per square
cm.(cm2) of the area of for example the skin and/or exposed
tissue to which the dosage amounts of the composition is to be
applied.
Thus, according to another aspect of the invention,
Applicants have provided topically applicable percutaneous
(intracutaneous) penetrating (best targeting the epidermis)
systemic independent acting (not acting essentially through the
blood) pharmaceutical compositions (combinations and
formulations) comprising a plurality of dosage amounts each
comprising, together with pharmaceutical excipients suitable for
topical application, a therapeutically effective (to treat and
to assist to resolve diseases and conditions of the skin and
exposed tissue (for example basal cell carcinoma, the
precancerous, often recurrent, actinic keratoses lesions, fungal
lesions, "liver" spots and like lesions (found for the most part
in the epidermis), squamous cell tumours, metastatic cancer of
the breast to the skin, malignancies and/or tumours in the skin
primary and metastatic melanoma in the skin, genital warts
(condyloma acuminata), cervical cancer, and HPV (Human Papilloma
Virus) including HPV of the cervix, psoriasis (both plaque-type
psoriasis and nail bed psoriasis), corns on the feet and hair
lossion the head of pregnant women), non-toxic (to the patient)
dosage amount of a drug for example which inhibits prostaglandin
synthesis, preferably a non-steroidal anti-inflammatory drug
(NSAID), for example, diclofenac, indomethacin, naproxen, and
(+/-) tromethamine salt of ketorolac (sold under the trademark
ToradolTM) and an effective non-toxic amount of hyaluronic acid
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and/or salts thereof (for example, the sodium salt) and/or
homologues, analogues, derivatives, complexes, esters,
fragments, and/or sub-units of hyaluronic acid ~preferably
hyaluronic acid and salts thereof) to transport (facilitate or
cause the transport of) the drug (for example NSAID's) rapidly
to the site in the skin (for example epidermis) and/or exposed
tissue of the disease or condition into the tissue to remain
there for a prolonged period of time to assist to treat and
~ssist to resolve the disease or condition for example by
blocking prostaglandin synthesis.
Effective dosage amounts of the form of hyaluronic
acid to facilitate or cause the transport of the drug into the
skin and/or exposed tissue by the form of hyaluronic acid
exceeds about 5 mg. - 10 mg. in the dosage amount administered
~applied and rubbed in) for each 1 cm~ of skin and/or exposed
tissue area of the disease or condition (for example basal cell
carcinoma) to which the dosage amount is applied. The dosage
amount applicable will depend upon the surface area of the skin
and/or exposed tissue in which the condition or disease exists.
Thus if the disease or condition occupies about .5 cm2, in
excess of about 2l/2 mg of the form of hyaluronic acid would be
used (applied and rubbed in). In the same way if the area is 2
cm2, the amount of the form of hyaluronic acid preferably
exce'eds about ! 10-20 mg of the dosage amount of the formulation
or composition applied. Preferred forms of the hyaluronic acid
(for example hyaluronic acid and the sodium salt thereof) have
molacular weights less than about 750,000 daltons (for example
about 150,000 to about 225,000 daltons) to transport the
medicine in the skin and/or exposed tissue. While higher
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molecular weights of the hyaluronic acid and forms thereof may
be used to penetrate the skin and/or exposed tissue and
transport the medicines or drugs, where the molecular weight of
the hyaluronic acid chosen for use is very large, it is
preferred that the form of hyaluronic acid is autoclaved, to
break down the hyaluronic acid to fragments of lesser molecular
weight or if feasible diluted to permit administration and
ensure no coagulation on or in the skin. Where the molecular
weight of the form of hyaluronic acid being employed is large,
the concentration of the form of the hyaluronic acid in the
composition may for example be reduced (for example to less than
about 3%) dependent on the molecular weight.
The blockage of prostaglandin synthesis by the
transported drug (for example NSAIDS) then unblocks the
macrophages and permits the macrophages of the patient proximate
the lesion (for example, the basal cell carcinoma) to destroy
the lesion or condition. Treatment by dosage amounts of the
composition (formulation and/or combination) eliminates the
condition without recurrence, even where the lesion has recurred
a number of times after unsuccessful treatments according to the
prior art.
Other non-steroidal anti-inflammatory drugs (NSAIDS)
may be used such as other propionic acid derivatives, Ibuprofen,
acetylsalicylic acid, piroxicam and flunixin.
When dosage amounts of such compositions, combinations
and formulations are applied to the site of the disease or
condition for example the basal cell carcinoma of the patient
suffering from the basal cell carcinoma, over a period of time
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(for example, for a period of 2-4 weeks 3 times daily) the basal
.~ ~
cell carcinoma is completely resolved and disappears.
Thus according to another aspect of the invention
there is provided a pharmaceutical composition from which dosage
amounts may be taken for application to the skin and/or exposed
tissue, the pharmaceutical composition comprising in a form for
application to a human a plurality of dosage amounts of medicine
and/or therapeutic agent to treat a disease or condition in a
human and a plurality of dosage amounts of hyaluronic acid
and/or salts and/or homologues, analogues, derivatives,
complexes, esters, fragments, and/or sub-units of hyaluronic
acid such that when dosage amounts of the pharmaceutical
composition are taken from the composition, the amount of the
medicine and/or therapeutic agent comprises an effective non-
toxic dosage amount of the medicine to treat the disease orcondition in the skin and/or exposed tissue in a human and the
amount of the form of hyaluronic acid in the dosage amount is
present in an effective amount to transport (facilitate or cause
the transport of) the medicine and/or therapeutic agent
intradermally (percutaneously, intercutaneously,
intracutaneously) into the skin (preferably to the epidermis
and dermis) and/or exposed tissue of a human to the site of a
pathology and/or trauma. The effective amount of the form of
` hyaluronic acid has a molecular weight and concentration to
transport the medicine ~drug) and/or therapeutic agent to the
site of trauma and/or pathology in the skin and/or exposed
tissue. In this regard the preferred amount of the form of
hyaluronic acid in each dosage amount exceeds 5 mg./cm2 and
preferably the molecular weight is less than about 750,000
8 ~
daltons, (in one embodiment about 150,000 to about 225,000
daltons) in some embodiments with a concentration of between
about 1 and 3%, preferably concentrations of between about 2 to
about 3% by weight. Where forms of hyaluronic acid are used
having greater molecular weights, they are preferably cleaved
and/or diluted to smaller concentrations, to facilitate or cause
the transport of the medicine and/or therapeutic agent.
According to another aspect of the invention there is
provided a pharmaceutical composition (for example a gel or
cream) from which dosage amounts may be taken and applied to the
skin to treat a disease or condition in humans, for example as
discussed above, the pharmaceutical composition comprising:
~ 1) a medicinal and/or therapeutic agent suitable
for treating a disease or condition in the skin and/or exposed
tissue in humans, for example a drug which inhibits
prostaglandin synthesis (for example an NS~ID); and
(2) hyaluronic acid and/or salts thereof and/or
homologues, analogues, derivatives, complexes, esters,
fragments, and sub-units of hyaluronic acid, in a form suitable
for administration to the skin and/or exposed tissue in humans;
characterized in that an effective non-toxic dosage amount
comprising components (1) and (2) taken and administered from
sald composition (i) is available in the skin and/or exposed
tissue upon administration to treat said disease or condition in
humans by penetration at the site to be treated ~o the site of
trauma and/or pathology, and (ii) comprises an effective non-
toxic dosage amount of component (2) effective to transport
~facilitate or cause the transport of) component (1) immediately
upon administration percutaneously into the skin (preferably the
~''~''':`' ' ''
- 22 - 2~
epidermis) to the site to be treated for example the site of
trauma and/or pathology where it remains for a prolonged time,
accumulating there and from which it is discharged via the
lymphatic system.
Therefore according to another aspect of the invention
a pharmaceutical composition is provided comprising~
(1) a medicinal and/or therapeutic agent which for
example inhibits prostaglandin synthesis in a therapeutically
effective amount to treat a disease or condition of the skin
and/or exposed tissue;
and (2) hyaluronic acid and/or salts thereof and/or
homologues, analogues, derivatives, complexes, esters,
fragments, and subunits of hyaluronic acid,
characterized in that said composition
(a) is in a dosage form (for example a gel or cream)
which is suitable for administration to the skin and/or exposed
tissue;
and (b) is in such an amount and in such form that
(i) component tl) is in an effective dosage amount to treat
said disease or condition by penetration at the site of the skin
and/or exposed tissue to be treated for example the basal cell
carinoma and other lesions; and (ii) component (2) is
immediately available to transport (facilitate or cause the
. transport of) component (1) to the site of trauma and/or
pathology to be treated, percutaneously into the skin (or
exposed tissue) where the composition resides and accumulates
for a prolonged period, and which component (2) is in an
effective non-toxic dosage amount to transport (facilitate or
cause the transport of) component (1) upon administration,
-~ - 23 - ~0~21
percutaneously into the skin or exposed tissue to the site of
the trauma and/or pathology. Preferably the form of hyaluronic
acid in the composition comprises hyaluronic acid and/or salts
thereof. An effective amount of the form of hyaluronic acid
exceeds about 5-10 mg per square centimeter (cm2) of skin and/or
exposed tissue to which it is to be applied.
According to another aspect of the invention there is
provided the use of:
(1) a medicinal and/or therapeutic agent for
example which inhibits prostaglandin synthesis,
and (2) hyaluronic acid and/or salts thereof and/or
homologues, analogues, derivatives, complexes, esters,
fragments, and subunits of hyaluronic acid,
in the manufacture of a pharmaceutical composition for
treating a disease or a condition (for example those discussed
above) of the skin and/or exposed tissue in a therapy wherein
. .
dosage amounts taken from the composition each comprise:
(1) a therapeutically effective amount of said
medicinal and/or therapeutic agent and
(2) a therapeutically effective amount of the ;
hyaluronic acid and/or salts thereof and/or homologues,
, :~., ::
analogues, derivatives, complexes, esters, fragments, and sub- ~;
units of hyaluronic acid, the pharmaceutical composition being
charlacterized in that for each dosage amount taken from the
25 pharmaceuti~al composition, the amount of component (2) is - ;
immediately available to transport component (1) percutaneously
to the site of trauma and/or pathology for example into the
epidermis where the composition accumulates and remains for a
prolonged period, at the site of the skin or exposed tissue to
-'"" ':', '
'. ' ''' ,"
2 ~ ~ 9 ~
be treated, and component (2) is in an effective non-toxic
amount to transport (facilitate or cause the transport of)
component (1) into the skin or exposed tissue (for example into
the epidermis). Preferably component (2) is hyaluronic acid
and/or salts thereof and preferably the dosage amount of
component (2) in the amount of the composition taken from the
composition (to be taken from the composition) and applied to
the skin or exposed tissue is a dose amount greater than about
S-10 mg per cm2 of skin and/or exposed tissue to which the
dosage amount is to be applied.
The pharmaceutical composition will normally include
pharmaceutically compatible excipients to provide a form for
ease of administration to the skin and/or exposed tissue for
transport into the epidermis. For example a suitable dosage
amount of a gel may be squeezed from a tube as a ribbon of gel
"X" cm long (which dosage amount (in the form of the ribbon "X"
cm long) contains the effective non-toxic dosage amounts of the
drug and form of hyaluronic acid. Or a dosage amount of cream
packaged in a jar may be scooped from the jar by a measuring
device or by "two fingers" in a suitable amount (for example in
a spoon containing a premeasured volume or an amount about half
the "length of the fingers"). Each of the dosage amounts
selected comprises the effective amounts of drug (for example
' NSAID) and effective amount of the form of hyaluronic acid (for
example hyaluronic acid and/or salts thereof). In this way the
patient may "squeeze" or "scoop" or "what have you" the
appropriate dosage amount and apply ~rub in) the dosage amount
onto the skin and/or exposed tissue for transport into the
epidermis.
- 25 - 2 ~ 21
,
Thus, according to another aspect of the invention, a
method of treating a disease and/or condition of the skin or
exposed tissue, for example basal cell carcinoma, the
precancerous, often recurrent, actinic keratoses lesions, fungal
lesions, "liver" spots and like lesions (found for the most part
in the epidermis), squamous cell tumours, metastatic cancer of
the breast to the skin, primary and metastatic melanoma in the
skin, malignancies and/or tumours in the skin, genital warts
(condyloma acuminata), cervical cancer, HPV (Human Papilloma
Virus) including HPV of the cervix, psoriasis ~both plaque-type
psoriasis and nail bed psoriasis), corns on the feet and hair
loss on the head of pregnant women, in a human is provided
comprising administering topically to human skin and/or exposed
tissue an effective non-toxic dosage amount of a composition
comprising, together with pharmaceutical excipients suitable for
topical application to the skin and/or exposed tissue, for
example in the form of a gel or cream (to give the composition
definition and form so that specific dosage amounts are easily
selected or taken for administration (for example squeezed from
a tube or scooped from a jar and rubbed into the skin or exposed
tissue), a therapeutically effective (to treat and to assist to
resolve the disease or condition for example basal cell
carcinoma or other lesion), non-toxic (to the patient) dosage
, i amount of a drug for example; which inhibits prostaglandin
synthesis, for example a non-steroidal anti-inflammatory drug
(NSAID), for example, diclofenac, indomethacin, naproxen, and
(+/-) tromethamine salt of ketorolac (sold under the trademark
ToradolTM) and an effective non-toxic dosage amount of
hyaluronic acid and/or salts thereof (for example, the sodium
~ - 26 ~ 2~39~21
salt) and/or homologues, analogues, derivatives, complexes,
esters, fragments, and/or sub-units of hyaluronic acid
(preferably hyaluronic acid and salts thereof) to transport
(facilitate or cause the transport of) the drug ~for example
S NSAID) into the skin or exposed tissue to the site of the
disease or condition to be treated percutaneously, (to the site
of trauma and/or pathology), for example into the epidermis,
where the form of hyaluronic acid and medicine accumulates and
remains for a prolonged period of time thereby for example
blocking prostaglandin synthesis in the skin or exposed tissue.
The form of hyaluronic acid is then cleared through the
lymphatics (lymphatics system).
Thus, according to another aspect of the invention,
the treatment may employ the use of the composition, formulation
or combination for the treatment of the diseases and conditions
aforesaid as for example by applying dosage amounts of the
composition, formulation or combination a number of times daily
(for example, 3 times daily) for a period of time, for example,
2-4 weeks to clear the disease, lesion or condition. Each
dosage amount applied will depend upon the size of the lesion or
condition on the skin or exposed tissue. For example, a
suitable dosage amount may include S-10 mg. of the form of
hyaluronic acid per 1 cm2 skin area or exposed tissue area.
One~ such formulation may `comprise 3% (by weight)
diclofenac in a 21/2~i (by weight) hyaluronic acid (sodium
hyaluronate - molecular weight 661,600) gel formulation, with
the excipients being glycerine (5%), benzyl alcohol (3~) (acting
in part as a solubilizer and preservative), and sterile water
(the balance) in a 50 gm. tube of the composition (a plurality
,. . , . . - ~ . . ~ . . . .
2~$9~21 ~
of dosage amounts) whose tube O.D. (outer diameter) of the
opening through which the gel formulation is discharged from the
tube is 8 mm and whose I.D. (inner diameter) of the opening is 4
mm. Therefore a ribbon 2-3 cm in length, squeezed from a tube
gives about 5 mg-7~/~ mg of hyaluronic acid for application to a
skin or exposed tissue surface area of 1-1 /7cm2 with an
effective dosage amount of diclofenac. While greater amounts
squeezed from the tube, may be applied, the application of
substantial excessive dosage amounts to the skin and/or exposed
tissue may saturate the skin or exposed tissue and thus the
epidermis. (There is therefore no more room for the composition
to pass between the cells and therefore further applications at
that time will not provide additional benefit). Where pain
relief is also required additional dosage amounts, for example
in excess of about 10 mg. of the hyaluronic acid taken from the
same pharmaceutical composition applied per/cm2 of surface area
: . .
of the skin or exposed tissue may be required to be applied.
Another formulation may comprise 3~ (by weight)
dlclofenac in a 2l/7% ~by weight) hyaluronic acid (sodium
hyaluronate - molecular weight 679,000) gel formulation (also in
a tube) with excipients being benzyl alcohol (1%) (a
preservative), methoxypolyethylene glycol 350 (20~ by weight) (a
solubilizer), and sterile water (the balance).
While the above compositions, combinations and
formulations are proposed, provided there is sufficient amounts
of the form of the hyaluronic acid (for example, sodium
hyaluronate) in the dosage amounts applied to the skin and/or ~;
exposed tissue to facilitate or cause the percutaneous
~intracutaneous) transport of the drug for example which
' .
- 208~21
inhibits prostaglandin synthesis, preferably an NSAID (for
example, diclofenac) to block prostaglandin synthesis, then the
formulations may be of any suitable form, for example, a 1
lotion of hyaluronic acid with NSAID, or a cream or gel or any
other suitable form.
Therefore according to another aspect of the
invention, there is provided containers ~for example tubes and
jars) containing compositions comprising a plurality of dosage
amounts of the drug and form of hyaluronic acid, each dosage
amount comprising an effective non-toxic dosage amount of the
drug and an effective non-toxic dosage amount of the form of
hyaluronic acid (preferably sodium hyaluronate having molecular
weight less than about 750,000 daltons) to transport the drug
into the skin and/or exposed tissue. In some embodiments, means
are provided to assist the removal from the container of an
effective dosage amount of the composition in the container for
use to apply to the skin or exposed tissue at the site of trauma
and/or pathology to treat the disease and/or condition ~for
example mouth opening of a tube to control the amount discharged
from the tube).
Furthermore, because there is little concern with
respect to the toxicity or adverse effects of the use of, for
example, the NSAIDs with the hyaluronic acid in the compositions
of this invention the NSAID may be combined as needed (after
solubilizing (if required) of the NSAID in a suitable
solubilizer) with the form of the hyaluronic acid.
Therefore according to another aspect of the
invention, percutaneous ~intercutanous) delivery of a
therapeutically effective dosage amount of a drug (in a
- 29 - 2
:
composition, combination or formulation) and which drug for
example inhibits prostaglandin synthesis, preferably being a
non-steroidal drug (NSAID) is provided. In this regard the drug
is transported to the site of, on, or in the skln and/or exposed
tissue of trauma and/or pathology to treat the disease or
condition for example the basal cell carcinoma or actinic
keratoses lesion in a mammal (human). The delivery may comprise
topically administering (to the skin or exposed tissue site of
:,: ',,
for example the basal cell carcinoma or other lesion) a
therapeutically effective non-toxic (to the patient) dosage
amount of a composition comprising a drug for example which
inhibits prostaglandin synthesis, preferably an NSAID (non-
steroidal anti-inflammatory drug), for example, diclofenac,
indomethacin, naproxen, and ~+/-) tromethamine salt of ketorolac
(sold under the trademark Toradolr~), and an effective non-toxic
amount of hyaluronic acid and/or salts thereof and/or
: .:
homologues, analogues, derivatives, complexes, esters,
fragments, and sub-units of hyaluronic acid, preferably
hyaluronic acid and salts thereof, sufflcient to transport,
(facilitate or cause the transport of), the drug for example
NSAID percutaneously (to for example the epidermis) to the site
of the trauma and~or pathology in for example the epidermis, for
example the basal cell carcinoma (or other lesion), to be
treated for example to block the synthesis of prostaglandins.
Delivery may be also accomplished by the same amount
of the form of hyaluronic acid, of other drugs percutaneously
(intercutaneously) to the skin and exposed tissue by application
and rubbing in of an effective non-toxic dosage amount of the
formulation or composition comprising an effective non-toxic
.: :
~ 30 ~ 2 ~
dosage amount of the drug and an effective non-toxic dosage
amount of the form of hyaluronic acid for the transport of the
drug percutaneously into the skin or exposed tissue to the
epidermis where the dosage amount of the composition is
accumulated and remains for a prolonged period of time before
the form of hyaluronic acid is cleared through the lymphatics.
In this regard the drug may be novantrone (an anti-cancer drug)
for administration to a tumour or malignancy in the skin. The
novantrone may comprise 10 mg in the dosage amount of the
composition and the form of hyaluronic acid may be in excess of
about 5 mg of sodium hyaluronic per cm2 of the skin or exposed
tissue (about 2. 59~i of the composition) for the percutaneous
transport of the novantrone.
Thus, according to another aspect of the invention,
use of a composition, combination or formulation is provided to
treat a disease or condition for example basal cell carcinoma
~or other lesion), by the application of the composition,
combination or formulation, the amount of the composition,
combination and formulation administered comprising together
with pharmaceutical excipients suitable for topical application,
a therapeutically effective (to treat and assist to resolve a
disease or condition for example, basal cell carcinoma), non-
toxic (to the patient) amount of a drug for example which
inhibits prostaglandin synthesis prèferably a non-stleroidal
anti-inflammatory drug (NSAID), for example, diclofenac,
indomethacin, naproxen, and (+/-) tromethamine salt of ketorolac
~sold under the trademark ToradolTM) administered together with,
or carried in, an effective dosage amount of hyaluronic acid
and/or salts thereof (for example, the sodium salt) and/or
2 ~
homologues, analogues, derivatives, complexes, esters,
fragments, and/or sub-units of hyaluronic acid (preferably
hyaluronic acid and salts thereof) effective to transport the
drug for example the NSAID (to facilitate or cause the transport
of the drug for example NSAID) percutaneously into the skin
especially the epidermis at the site of the disease or condition
for example basal cell carcinoma (or other lesion) to be
treated, thereby, if an NSAID, blocking prostaglandin synthesis
to enable the macrophages (and N.K. cells) to resolve the
disease or condition for example basal cell carcinoma or other
lesion.
Applicants postulate that the hyaluronic acid and/or
salts thereof and/or the homologues, analogues, derivatives.,
complexes, esters, fragments, and/or sub units of hyaluronic
acid facilitate or cause the transport of the drug for example
which blocks prostaglandin synthesis (preferably an NSAID) to
the site of prostaglandin synthesis to block prostaglandin
synthesis.
Applicants' compositions and dosage amounts of their
compositions and the use of their compositions and dosage
amounts of their compositions, at the same time, abate pain that
the patient is experiencing at the paccinian nerve bundles
(superficial nerve bundles) at the site of the trauma and/or
pathology on/in the exposed tissue and/or skin.
Thus, according to another aspect of the invention, a
method of abating pain in the skin and/or exposed tissue for
example suffering a disease or condition (for example those
discussed above), and a composition from which dosage amounts
may be taken and applied (rubbed in) which is useful for abating
- 32 ~ 2 ~3~ ~2 1
,
such pain are provided, the method comprising administering
(rubbing on) an effective dosage amount of the composition to
the skin and/or exposed tissue, and the composition comprises a
plurality of dosage amounts, each comprising an effective non-
toxic dosage amount of an NSAID and an effective non-toxic
dosage amount of the hyaluronic acid and/or salts thereof and/or
homologues, analogues, derivatives, complexes, esters,
fragments, and/or subunits of hyaluronic acid (preferably
hyaluronic acid and salts thereof), for example amounts
exceeding 10-20 mg. per square cm (cm~) of skin or exposed
tissue to which it is applied, for percutaneous transport of the
NSAID by the form of hyaluronic acid into the epidermis
proximate the paccinian nerve bundles (superficial nerve bundles
at the end of the nerves) to abate pain. Thus, according to
another aspect of the invention, compositions are provided for
use to relieve pain from which dosage amounts of the composition
comprising dosage amounts of the NSAID and form of hyaluronic
acid are taken.
By way of example and to illustrate the facilitation
of the delivery or transport of a chemical to a site in a human,
when ethyl alcohol is injected directly into a tumour and
sonographic (ultrasound) assessment is made, it is not dispersed
throughout the tumour. When the ethyl alcohol to be
administered into a tumour is carried by hyaluronic acid and/or
salts thereof, sonographic assessment of the tumour demonstrates
the dispersion of the ethyl alcohol throughout the tumour.
While Applicants postulate that the hyaluronic acid
facilitate or causes the transport and delivery, Applicants'
invention may be used as described irrespective of the actual
,
2~8~2~
. ::
method of operation of the hyaluronic acid and/or salts thereof
and/or the homologues, analogues, derivatives, complexes,
esters, fragments and sub-units of hyaluronic acid.
The combination of hyaluronic acid and salts thereof
and other forms with drugs for example that inhibit
prostaglandin synthesis, for example NSAIDs, alters their
distribution and performance in the skin and/or exposed tissue
particularly the epidermis (the comblnations and formulations
being systemic independent), and produces an unusual targeting
for underperfused skin and/or pathological tissue in the skin
~site of trauma and/or pathology). The application may be made
as re~uired with the amount depending upon the condition of the
skin or exposed tissue.
As a major amount of soluble indomethacin may be
incorporated into the formulation, or composition, the
indomethacin may be solubilized using n-methyl glucamine at a
dilution of 5mg/ml of n-methyl glucamine (NMG). This substance
is then passed through a 22 micron Milipore filter to produce
sterility. This material is non-toxic at 16 fold the
therapeutic dose in animals (with hyaluronic acid) and for this
reason was considered appropriate to be used in human
conditions. Thus, Indocid~M solubilized in NMG may be
administered with hyaluronic acid topically for percutaneous
penetration at, for example, varying doses. The solution of
indomethacin and NMG may be mixed with, for example,
"LifeCoreTM" hyaluronic acid in dosage amounts discussed above.
This produces an appropriate mixture and can be administered
safely.
' ',: '~ '
- 34 ~ 2~9~21
When the NSAID, for example indomethacin (dissolved in
n-methyl glucamine) or other NSAID, is applied topically in an
effective dosage amount from a composition or formulation also
including the effective dosage amount of the form of hyaluronic
acid, no major toxic side effects occur, such as gastro-
intestinal distress, neurological abnormalities, depression,
etc., even at elevated amounts of indomethacin (if necessary).
(This may be in part because of the clearing of the hyaluronic
acid through the lymphatic system from the site). In addition,
the responses that have been observed are dramatic when the drug
for example NSAID (for example diclofenac) is combined with
hyaluronic acid, demonstrating clearly that the combination is
now "targeting" to the site of pathology or trauma, or
pathological tissue. Furthermore, patients using the
formulations and combinations of drug (for example NSAID)
hyaluronic acid ~sodium hyaluronate) (for example, diclofenac or
indomethacin and hyaluronic acid), experience dramatic relief of
pain immediately.
Thus, Applicants believe that the use of the NSAID,
for example with hyaluronic acid (sodium hyaluronate), deblocks
the macrophages (and N.K. cells (Natural Killer Cells) thought
to be immature macrophages) by preventing enzymatic production
of prostaglandin which blocks macrophage (and N.K. cell)
functioning. The hyaluronic acid (and salt and other forms) not
only enhances the activity of the drug (NSAID) but also reduces
any side effects and toxicity that is associated with the use of
the prostaglandin synthesis inhibitors. When effective dosage
amounts of compositions, formulations and combinations
containing effective dosage amounts of the drugs for example,
. , . ,:
2 ~
- 35 -
(NSAIDs (for example, diclofenac)) and effective dosage amounts
of, for example, hyaluronic acid or the sodium salt thereof, are
applied to for example the tumour lesion (for example basal cell
carcinoma) or other condition (for example, actinic keratoses
lesion) for a period of time (for example, 3 times daily for 2-4
weeks), the carcinoma and lesions, as the case may be,
disappear.
Applicants also postulate that when the combination or
formulation is applied to the disease or condition (for example,
basal cell carcinoma or actinic keratoses), the hyaluronic acid
passes between the cells (in the stratum corneum and epidermis
to the dermis depending on amounts) to the areas of trauma
and/or pathology deficient in hyaluronic acid (or forms
thereof), transporting, taking, drawing, carrying or pulling the
NSAID with it to the sites of prostaglandin synthesis,
penetrating to inhibit prostaglandin synthesis until the space
between the cells is saturated. The NSAID now being proximate
the Paccinian nerve bundle (superficial nerve bundles at the end
of the nerves) gives pain relief. The macrophages (which had
been previously blocked) are unblocked and act to destroy the
disease or condition for example basal cell carcinoma, actinic
keratoses leslon, or other disease or lesion. Furthermore, the
effective non-toxic dosage amount of the composition,
combination or formulation, comprising the effective dosage
amount of the form of hyaluronic acid and the effective dosage
amount of NSAID passing through the stratum corneum to the
epidermis and to the dermis (if a sufficient amount of the form
of hyaluronic acid is present), passes into the skin,
accumulating and staying longer in the skin at the site of the
:
'', '
- 36 ~ 2~
trauma and/or pathology. Therefore, after having had an
immediate effect at the site of trauma and/or pathology (for
example, relieving pain and acting on the basal cell carcinoma,
actinic keratoses and other disease, condition or lesion), the
NSAID-hyaluronic acid combination continues to accumulate at the
site in need of treatment and thereafter clears through the
lymphatic system.
Thus according to another aspect of Applicant's
invention, Applicants' compositions, formulations and
combinations quickly penetrate on application through the
stratum corneum into the epidermis (to the dermis) by the form
of hyaluronic acid transporting the NSAID, to the site of trauma
and/or pathology where the amounts applied accumulate and remain
for a prolonged time for treatment.
Fifteen (15) minutes after application of one of
Applicants' formulations, about three times the amount of
Applicants' formulation has penetrated into the skin
(particularly the epidermis) than formulations and combinations
not containing hyaluronic acid or effective dosage amounts of
hyaluronic acid, but containing the same drug. Furthermore, the
drug and hyaluronic acid accumulate and remain at the site in
need of treatment for a longer period of time.
Thus according to another aspect of the invention,
non-toxic effective dosage amounts of forms of hyaluronic acid
(preferably sodium hyaluronate) and effective non-toxic dosage
amounts of a drug may be administered in compositions to sites
of trauma or pathology, on/in the skin and/or exposed tissue
(for example the epidermis) by the application of the effective
non-toxic dosage amount of the composition comprising an
'
: ' ,
2~3~21 :
effective non-toxic dosage amount of a drug ~for example an
NSAID) and an effective non-toxic dosage amount of a form of
hyaluronic acid (for example sodium hyaluronate) to the skin or
exposed tissue whereby the forms hyaluronic acid transport the
drug percutaneously to the site of trauma and/or pathology where
the composition accumulates and remains for a prolonged period
of time thereby retaining the drug at the site of trauma and/or
pathology (for example the epidermis) for the treatment of the
condition or disease and the reduction of pain. :
..
Thus according to another aspect of the invention,
Applicants have provided compositions (formulations and
combinations) (including pharmaceutical excipients suitable for
topical application) from which effective non-toxic (to the
patient) dosage amounts of a drug (for example an NSAID) to
treat and to assist to resolve diseases and conditions of the
skln and/or exposed tissue (for example basal cell carcinoma,
the precancerous, often recurrent, actinic keratoses lesions,
fungal lesions, "liver" spots and like lesions (found for the
most part in the epidermis), squamous cell tumours, metastatic
cancer of the breast to the skin, primary and metastatic
melanoma in the skin, malignancies and/or tumours of the skin,
gential warts, cervical cancer, and HPV (Human Papilloma Virus)
including HPV of the cervix, psoriasis (both plaque-type
psoriasis and nail bed psoriasis), corns on the feet and hair
loss on the head of pregnant women), and effective non-toxic
dosage amounts of hyaluronic acid and/or salts thereof (for
example, the sodium salt) and/or homologues, analogues,
derivatives, complexes, esters, fragments, and/or sub-units of
hyaluronic acid (preferably hyaluronic acid and salts thereof)
'". . ::,;',
- 2 ~
sufficient to transport (to facilitate or cause the transport
of) the drug, for example NSAID, are taken for application, to a
site in the skin (for example epidermis) or exposed tissue
having a disease or condition for percutaneous transport into
the skin and/or exposed tissue to accumulate and remain there
for a prolonged period of time to for example block
prostaglandin synthesis. Thus an effective dosage amount of the
composition or formulation or combination penetrates quickly
into the skin, for example by the hyaluronic acid transporting
the NSAID or causing the NSAID to be transported for example to
the epidermis of the skin, accumulates there and remains there
for a prolonged period of time, thereby accumulating the drug
and forms of hyaluronic acid in the skin (particularly the
epidermis).
Thus according to another aspect of the invention, a
method of accumulating a drug and a form of hyaluronic acid in
skin and/or exposed tissue is provided comprising topically
..
administering a therapeutically effective non-toxic dosage
amount of a composition comprising pharmaceutical excipients
20 suitable for topical applications, an effective non-toxic (to :
the patient) dosage amount of a drug for example which inhibits `
prostaglandin synthesis, preferably a non-steroidal anti- ;
inflammatory drug ~NSAID), for example, diclofenac,
indomethacin, naproxen, and (~/-) tromethamine salt of ketorolac ~ :
25 (sold under the trademark Toradolr~) (to treat and to assist to ~:
resolve the disease and conditions of the skin and exposed
tissue (for example basal cell carcinoma, the precancerous, :~;
often recurrent, actinic keratoses lesions, fungal lesions, .;
"liver" spots and like lesions (found for the most part in the : .
' ',.'. ~":,'
- 39 -
~``` 20~9~%1 ~ ~
epidermis), squamous cell tumours, metastatic cancer of the
breast to the skin, malignancies and/or tumours of the skin, :~
primary and metastatic melanoma in the skin, genital warts
cervical cancer, and HPV (Human Papilloma virus) including HPV
of the cervix, psoriasis (both plaque-type psoriasis and nail
bed psoriasis), corns on the feet and hair loss on the head of ~-
.
pregnant women), and an effective non-toxic dosage amount of `~
hyaluronic acid and/or salts thereof (for example, the sodium
salt) and/or homologues, analogues, derivatives, complexes, :
esters, fragments, and/or sub-units of hyaluronic acid
(preferably hyaluronic acid and salts thereof) effective to
transport ~to facilitate or cause the transport of) the drug
(for example NSAID) percutaneously to the site in the skin (for
example epidermis) or exposed tissue of the disease or condition : i;
15 to accumulate and remain there for a prolonged period of time :;:
for example to block prostaglandin synthesis. ~; .
According to another aspect of the invention, a method ~ i.
. . .. ~.
of quickly delivering a drug to the skin or exposed tissue,
particularly the epidermis, and maintaining the drug therein for
a prolonged period of time is provided, the method comprising
topically administering (for example rubbing in) an effective
non-toxic dosage amount of a composition comprising
pharmaceutical excipients suitable for topical application, a
therapeutically effective (to treat and assist to resolve the
.
25 disease and/or condition of the skin and exposed tissue (for ~ :
example basal cell carcinoma, the precancerous, often recurrent,
actinic keratoses lesions, fungal lesions, "liver" spots and
like lesions (found for the most part in the epidermis),
s~uamous cell tumours, metastatic cancer of the breast to the
- 40 -
2089621
skin, primary and metastatic melanoma in the skin, malignancies
and/or tumours of the skin, genital warts, cervical cancer, and
HPV (Human Papilloma Virus) including HPV of the cervix,
psoriasis (both plaque-type psoriasis and nail bed psoriasis),
corns on the feet and hair loss on the head of pregnant women)),
non-toxic (to the patient) dosage amount of a drug for example
which inhibits prostaglandin synthesis, preferably a non-
steroidal anti-inflammatory drug (NSAID), for example,
diclofenac, indomethacin, naproxen, and (+/-) tromethamine salt
of ketorolac (sold under the trademark Toradoll~) and an
effective non~toxic dosage amount of hyaluronic acld and/or
salts thereof (for example, the sodium salt) and/or homologues,
analogues, derivatives, complexes, esters, fragments, and/or
sub-units of hyaluronic acid (preferably hyaluronic acid and
salts thereof) sufficient to transport (to facilitate or cause
the transport of) the drug for example the NSAID percutaneously
to the site of the trauma and/or pathology in the skin (for
example epidermis) or exposed tissue, for remaining there for a
prolonged period of time (for example in the epidermis and
dermis) to for example block prostaglandin synthesis. Suitable
amounts of the form of hyaluronic acid may comprise in excess of
5 mg. per cm2 in a form which transports the drug (for example
molecular weights of the form of hyaluronic acid being less than
about 750,000 Daltons or if at substantially greater molecular
weights, diluted (to reduce) the concentration or autoclaved or
cleaved if required to reduce the size of the molecules.
According to another aspect of the invention, a method
of controlling the unloading of a drug from the skin or exposed
tissue into the lymphatic system comprises delivering
208~6~
. . ~ .
(transporting) an amount of drug into the skin or exposed tissue
by an effective non-toxic dosage amount of a form of hyaluronic
acid and/or salts thereof and/or homologues, analogues,
derivatives, complexes, esters, fragments, and/or sub-units of
hyaluronic acid to the skin (epidermis) or exposed tissue to
control the unloading of the drug into the lymphatic system
(for example by the application of greater than 5 mg./cm2) of
the form of hyaluronic acid.
Thus according to another aspect of the invention a
composition is provided which when administered to a human by
preferably administration to the skin and/or exposed tissue of a
human, unloads its contents into the lymphatic system, the
composition comprising an effective non-toxic dosage amount of a
drug (for example an NSAID or an anti-cancer drug (Novantrone)
and an effective non-toxic amount of hyaluronic acid and/or
salts thereof and/or homologues, analogues, derivatives,
complexes, esters, fragments and/or sub-units of hya-luronic acid
(for example at least about 5-10 mg/cm~ of skin or exposed
tlssue). Thus the composition is made up of a plurality of such
dosage forms (for example a cream or lotion or gel).
Thus according to another aspect of the invention, a
new composition for treating diseases via the lymphatic system
is provided comprising a plurality of effective non-toxic dosage
amounts of the composition, each dosage amount comprising
hyaluronic acid and/or salts thereof and/or homologues,
analogues, derivatives, complexes, esters, fragments and/or sub-
units of hyaluronic acid for passing into the lymphatic system
and a therapeutic effective amount of medicine for treatment of
a disease (which disease may be in the lymphatic system).
- 42 -
~ 2~9~
According to another aspect of the invention, the
composition may be for application to the skin or exposed
tissue.
According to another aspect of the invention, a
composition is provided from which effective dosage amounts may
be taken and administered, each effective dosage amount of the
composition comprising an effective non-to~ic dosage amount of
hyaluronic acid and/or salts thereof and/or homologues, ~ ~;
analogues, derivatives, complexes, esters, fragments and/or sub~
units for transporting a therapeutically effective non-toxic
dosage amount of a medicine and/or therapeutic agent (for
example an NSAID) in the composition into the skin and/or
exposed tissue when applied thereto to an area of pathology
and/or trauma then into the lymphatic system, the dosage amount
belng essentially systemic independent such that substantial
amounts do not enter the blood system prior to clearing - ~
(passing) into the lymphatic system. ereferably the amount of ~` ;
the form of hyaluronic acid in each dosage amount administered
ls greater than about 5-10 mg./cm'` and the molecular weight is
20 less than about 750,000 daltons. ~
We have compared the penetration and retention of one ;
of our combinations (formulations) with a control and Voltarol
Emulgel in the skin as follows: ~:
.,~., .: '
- 43 -
2 ~ 2 1
(A) OUR FO~MULATION :.
, ,
~ ,~
1% DICLOFENAC IN 3.0~ HA GEL 50a/tube
EPDICLO1 ~
LOT XPB 044 Quantity 1500ml ;
FCRMVLA SupPlier Lot Amount Percent
Sterile Water Baxter AW45F1 1397ml -- ;
Glycerin Life 1043 95q(36ml) 3% ~ ~
10 Benzyl Alcohol Caledon 02517 22.5g(22ml) 1.5~ ~ :
Liquid Wax DICDD Brooks 191-175 45g 3% .
Diclofenac Sodium Proslntex 9113003 15g 1% ~:
Sodium Hyaluronate Skymart HG-1103 45g 3%
Mol. Wt. 661,600 . ::~
PROCEDURE
- Set up stirring apparatus using a 3 liter stainless steel
beaker
- Add Water, Glycerin, Benzyl Alcohol and Liquid Wax DICDD,
stir and mix for 10 minutes
- Add Diclofenac Sodium and stir for 30 minutes to dissolve
- Add Sodium Hyaluronate and stir for 90 minutes
E~E~
0 In a 50 ml aluminum collapsible tube,
inside of tube lacquered with a phanolic resin, outside of
tube white regular enamel coating;
9 mm white polypropylene screw on cap with pierce tip
Gels Batch No.s
35 ~B) Voltarol Emulgel 060400 10 93
tC) 1% Diclofenac Gel XPBO49 (Control)
2 ~ 3 9 6 ~
, .,
(C) CONTROL
1~, DICLOFENAC IN CA~APOL GEL 50a Jar k~ -
LOT XPB 049 Quantity 100ml ~ :~
. ::
5 FORMULA Supplier Lot Amount Percent
Sterile Water Baxter AW45N5 93ml
Glycerin BDH 2579 3g 3%
Benzyl Alcohol BDH 23797 1.5g 1.5%
Liquid Wax DICDD Brooks L-1424 3g 3% . --~:
10 Diclofenac Sodium Prosintex 9113003 lg 1% -.~-
Carbopol 934 A&C Chemicals 910304 lg 1% ` .. ..
- Set up stirring apparatus using a 400ml stainless steel
beaker .:- .;~
' " . ' `':
- Add Water, Glycerin, Benzyl Alcohol, Liquid Wax DICDD, and
stir to mix thoroughly for 10 minutes ~;
- Add Diclofenac Sodium and stir for 20 minutes to dissolve :~
-: :.
- Very slowly add Carbopol 934, avoid getting lumps : ~
Samples .:~.
Cell Sample Quantity of gel applied
(mg) :.;~
25 A 060400 10 93 192
B 060400 10 93 192 ; ;
C EPDICLO1* 192
D EPDICLO1* 192 . ;
E XPB049 192
30 F XPB049 192
* - Our Formulation . :~ :
Skin Type
One piece of skin (Female, 37 years, smoker, breast ; :
skin) was used for one sample from each batch. A second piece
.~ ...
35 of skin (no further details available) was used for the second ~ :
:'" ~'''
- 45 - 20$~
;:
sample from each batch. The skin was stored deep frozen (<-
20C) until thawed for this experiment. FU11 thickness skin was
used for this experiment. ~-
Experimental Conditions
Skin permeation cells were prepared containing an
exposed skin surface area of 9.6 cm2 and a constantly stirred
receptor fluid beneath the skin consisting of 135 ml of
ethanol:phosphate buffered saline (25:75 v/v).
Each cell was allowed to equilibrate for 1 hour at
37C after which the gel was spread evenly over the skin surface
at a concentration of 20 mg/cm2). See table above.
The cell was then maintained at 37C with an air temperature
above the skin of 35C.
24 hours after application of the gel the experiment
was stopped and a portion of the receptor fluid removed. The
skin was removed from the cell and any gel remaining on the
surface carefully wiped off with dry paper towel followed by
paper towel moistened with water. The skin was cut with a
scalpel to obtain thin top and thicker lower sections of skin.
This was done in order to obtain layers of skin which
approximated the epidermal and dermal layers. Each skln section
was weighed and the residual diclofenac extracted with lOml of
fresh receptor fluid using an ultra turrax homogeniser. The
homogenates were centrifuged and a portion of the resultant
supernatant solutions removed.
The receptor fluid and skin extracts from each cell
were assayed for diclofenac content by using a validated reverse
phase high performance liquid chromatography (HPLC) method.
,~
- 46 - ~
2~3~2~
Results -
Distribution of Diclofenaç 24 hours af~er application of - -:
Diclofenac Gel ~:
Samp~e _R~ce~or Top S~L~ _eQL~Q~ a~Om akin portion
~g Skin ~5 ~g/g Skin ~5 ~/g ~:
_ __ Weight _ Weight
~Voltarol . -~
Emug~l)
060400 10 93 447 0.1363 101 7421.244217 174 -
060400 10 93 764 0.2445 141 5771.235202 164
Mean 606 660 169 ~ ~
Formulation) .. ~ :
20 li~PDICLOl 247 0.1535 133 ô671.466148 101
ICPDICLOl 292 O .1647 145 ô 791.00286 86
Mean 269 873 93
~Control)
XP~049 184 0.1275 35 2721.1325O 51 '
XPB049 147 0.2068 82 3961.08968 63 ;
Mean 165 334 57
30 _ _ _ _ ' . :
Thus having regard to the above and Figures 1', 2' and .
3', it iS clear that the sodium hyaluronate takes the diclofenac :
into the skin to the epidermis level (See Figure 1') more
rapidly than the Voltarol Emugel or non-hyaluronic acid :
35 diclofenac containing control formulation, accumulates it there ~:
and retains it there longer. The other formulations permit the
NSAID, diclofenac, to pass through the bottom skin portion
(dermis) quicker, thereby clearing it from the epidermis and ;
' . . ~. ,~
.
203~21
dermis, quicker. Furthermore, more of Applicants' formulation
is in the epidermis and in the dermis even after 12 hours.
It is also clear that Applicants' formulations clear
into the lymphatic system not through the blood system. Yet the
5 prior art topical formulations have always tried "to drive" the
formulations through the skin into the blood for treatment of
the disease or condition in the area (i.e. systemic action).
Thus, our composition, formulation and combination,
(and dosage amounts thereof) penetrate quickly and rapidly at
the site of treatment through the upper skin into the epidermis,
where the paccinian bundles are located and the NSAID and the
form of hyaluronic acid are accumulated and are retained longer,
where needed (for example for the treatment of basal cell
carcinoma).
Further, the NSAIDs are retained in the area to be
treated with the form of hyaluronic acid. In doing so, they
preclude prostaglandin synthesis , in effect, deactivating the
synthesis or inhibiting the synthesis, of prostaglandins,
permitting the macrophages' scavenger cell activity to eliminate
the tumour and lesion. Additionally, a rapid onset of pain
relief (analgesic effect) is provided (depending on the amount
of NSAID and form of hyaluronic acid) usually where in excess of
about 10 mg of the form of hyaluronic acid (preferably
hyaluronic acid and salts thereof) is admini.stered per cm2 of
surface area comprises the dosage amount administered. However,
there are no blood levels of the NSAID in the immediate area of
treatment. The forms of hyaluronic acid are thus cleared via
the lymphatic system. Then the lymphatics pass the forms of
hyaluronic acid, Applicants believe, to the blood system. Thus,
- 48 ~ 2~ 21
. ~
the NSAIDs and forms of hyaluronic acid stay at the site to be
treated for well in excess of 12 - 24 hours, a protracted stay.
Thus, over the period of treatment (for example,
applications of effective non-toxic dosage amounts of
compositions containing for example effective non-toxic dosage
amounts of the NSAIDS and effective non-toxic dosage amounts of
the sodium hyaluronate, 3 times a day for 2-4 weeks, transport
the NSAIDS to to the epidermis to inhibit prostaglandin
synthesis to enable the macrophages to "scavenge" the tumour
cells and eliminate them. The end result is the successful
treatment of the disease or condition at the site of trauma
and/or pathology of the skin or exposed tissue, for example, the
resolution of, the basal cell carcinoma, the precancerous, often
recurrent, actinic keratoses lesions, fungal lesions, "liver"
spots and like lesions (found for the most part in the
epidermis)~ squamous cell tumours, metastatic cancer of the
breast to the skin, malignancies and/or tumours in the skin,
primary and metastatic melanoma in the skin, genital warts
cervical cancer, and HPV (Human Papilloma Virus) including HPV
Of the cervix, psoriasis (both plaque-type psoriasis and nail
bed psoriasis), corns on the feet and hair loss on the head of
pregnant women, with complete disappearance of the disease or
condition as the case may be, by topical therapy without
resorting to surgery.
One of the formulations which we have employed
successfully is a gel formulation comprising 3~ diclofenac in
2.5~ sodium hyaluronate formulated as follows:
,' ~ ,,
~'-
~ 49 ~ ~ ~8~2~
Formulation 1 (3000 ml.)
Formula Supplier (LOT) Amount Percent
Glycerine Life 1043150 g (119 ml) 5
Benzyl Alcohol Caledon 0251790 g (86 ml) 3
5 Diclofenac Sodium Prosintex 9113003 90 grams 3
Sodium Hyaluronate SkymarkHG1003 75 grams 2.
(MW 661,660)
Sterile water Baxter AW4~552795 ml.
balance
Procedure
- set up stirring apparatus using a 4 litre stainless
steel beaker
- add water, Glycerine, and Benzyl Alcohol; stir to mix
- add Diclofenac Sodium and stir for 30 minutes
- then add the Sodium Hyaluronate and stir for gO minutes
- initially, stir at a high torque but avoid splashing; as
the gel thickens, stir at a lower torque.
The gel is then packaged in a tube or jar or other
suitable container for use. Identification of suitable dosage
amounts and how they are taken from the container may be
provided with the container - for example squeeze "X" cm. of
ribbon from the tube; fill spoon or spatula accompanying jar;
~the spoon or spatula containing a predetermined dosage amount)
then apply and rub into site of trauma and/or pathology (the
dosage amount indicated will be such amount of the composition
whichlcomprises in excess of about 5 mg. of sodium hyaluronate
per cm2 ~square centimeter) of skin or exposed tissue to which
the dosage amount is to be applied. The amount of Diclofenac
Sodium was determined in the same manner (having regard to the
dosage amount required).
Another such formulation is:
~0~21
Formulation 2
Formula Sup~lier (LOT) Amount P~rcent
. :
Methoxypolyethylene Sigma 34F-0266 300 g. 20
Glycol 350
Benzyl Alcohol BDH 23797 15 g. 1 ~
Diclofenac Sodium Prosintex 9123012 45 g. 3 :
Sodium Hyaluronate Skymart HG 1009 37.5 g. 2.5
10 (MW 679,000) ~ :-
Sterile Water Baxter AW45R6 1200 ml.
balance
Procedure
15 - set up stirring apparatus using a 3 litre stainless steel :
beaker :
- add water, Metho~ypolyethylene Glycol 350, and Benzyl
Alcohol and stir for 20 minutes to mix ~:
- add Diclofenac Sodium and stir for 30 minutes to dissolve
- add Hyaluronate Sodium slowly and stir initially at a high
spèed, but avoid splashing
- after addition, stir at a slower speed for 90 minutes; the
slower speed reduces the formation of air bubbles
- the result is a clear, transparent, viscous gel which is
2S put into a container. Once again instructions are given
$or administration and if applicable measuring devices (to
provide a premeasured dosage amount) accompany the
container. ~
Still other formulations are: :
- 51 ~ f?~f~f~
,_ ~
Formulation 3
3% Diclofenac in 2.53~ HA Gel
Formula Supplier LOT Amount Percent
Sterile Water Baxter AW45K6 1200 ml
5 Methoxypolyethylene Sigma 34~-0266 300G (273 ml) 20%
Glycol 350
Benzyl Alcohol BDH 23797 15G (14 ml) lf~ ~
Diclofenac Sodium Prosintex 9123012 45 g 3% .
Sodium Hyaluronate Skymart HG 1004 37.5 g 2.5%
10 MW 679,000 :
Procef~lr~
':: .
15 - Set up stirring apparatus using a 2 liter stainless steel :~:
beaker, :~
- Add water, Methoxypolyethylene Glycol 350, and Benzyl
Alcohol and stir for 20 minutes to mix, ~
- Add Diclofenoc Sodium and stir for 30 minutes to disolve, . .~.
20 - Add Hyularonate Sodium slowly and stir initially at a high
speed, but avoid splashing,
- After addition, stir at a slower speed for 90 minutes,the :~
slower speed reduces the formation of air bubbles, ~ :
- The result is a clear transparent, viscous gel which is
poured into jars and tubes. Once again instructions
accompany the container and where applicable appropriate
devices for providing a premeasured amount of the -.
f;
composition accompany the container.
,~',:''- :''
.,' ~ :: :
~: ' ' ~", "j! '
- 52 - 2~ 21 ~
,
. .
.
Formulation 4 -~
S~ IBUPROFEN IN 3.0~ HA GEL, 50 ml JAR
Fo~mula Supplier LOT Amount Percent :;
Sterile Water saxter AW45R6 196 ml --
Meglumine Falk 15684 11 g 5. 5%
Ibuprofen BDH 19/241 10 g 5%
Benzy Alcohol BDH 23797 2 g 1%
Glycerin BD~ 2579 2 g 1%
10 Hyaluronate
Sodium Skymart HG 1003 6 g 3%
Mol Wt 661,600
15 PROCEDUBE :. `
- Set up stirring apparatus using a 300 ml stainless
steel beaker,
- Add Sterile Water and Meglumine, and stir for 10 minutes, ;
- Add Ibuprofen and stir for 15 minutes, :-~
- Add Benzyl Alcohol, followed by Glycerin and stir for 15
minutes,
- Finally, add Hyaluronate Sodium slowly and stir initially :~
at a high torque to mix, but avoid splashing, ~: .
- As the gel thickens, stir at a slow speed for 90 minutes.
,
, i i , ' ;
- 53 ~ 2Q~9~21
.,
Formulation 5
2~ PIROXICAM IN 2.5% HA GEL :~
Formula Supplier LOT Amount Percent
Sterile Water Baxter AW45R6 200 ml
Meglumine Falk 15684 8 g 4%
Piroxicam AMSA 1-010 4 g 2% - -
Hyaluronate Sodium Skymart HG 1003 5 g 2.5% :
MW 661,600 ~;
10 PROCEDURE :`
, ~.
- Set up stirring apparatus using a 300 ml stainless steel ~ ;
beaker,
- Add 200 ml of sterile water, ;:~
- Add 8 grams of Meglumine and dissolve,
- Very slowly add 4 grams of Piroxicam and stir for 20
minutes, ; :
- Slowly add 5 grams of Hyaluronate Sodium and stir at high ~;~
speed, .
- Stir for 90 minutes at a slower speed
COMMENTS
- A clear yellowish transparent gel
'"' ' '
. .
~ 54 ~ 2~9~21
r~~
Formulation 6 :
5% IBUPROFEN CREAM 50 ml JAR
.: .
OILY PHASE :
Formula Supplier LOT Amount ~ercent
Liquid wax DICDD ~rooks L-1424 450 g 15% . : .
~rookswax D Brooks P-490 480 ~ 16
Glycerin ~DH 109109/2578 153 ~(119 ml~ 5~ ~
AQUEOUS PHASE : ::
Sterile Water Baxter AW45F1 1950 ml -- ~-
Meglumine ~alk 15684 150 g 5
Ibuprofen BXH 19/241 150 g 5~
lS MW 200,00 - ~`
~P~i~D~y~ onate Skymart O01 4~ 5
~L~ervative Su~Qcidç-A Sutton SH-107 9 ~ _ o-3
: ' ,'
PROCEDURE
A - Add all the ingredients of the oily phase A into a 4 liter ;~
stainless steel beaker, melt at 55"c, finally heat to 75% ~ ~.
when Aqueous Phase B is ready .
B - Into a 3 liter stainless steel beaker, add 1950 ml water,
set up, the stirring apparatus, add the Me~lumine, stir to ~ .dissolve for 10 minutes,
- Slowly add Ibuprofen, stir to dissolve for 20 minutes, ;
- Very slowly add Sodium Hyaluronate and stir for one hour to ;
dissolve all the Sodium Hyaluronate,
- Finally, heat to 75C,with stirring for a total time of 30
minutes. ~
,. :'"
- 55 ~ 2~ 21
, ~
POUR B INTO A, both at a temperature of 75"C, slowly
- Remove the heat source and stir with a strong vortex for
one hour, ~;
- When the temperature has cooled down to 45C add ` -
preservative Suttocide A,
- Continue stirring at a slower speed until the temperature
is 35C,
- At 35C remove the propeller, pour into 50 ml jars. :
Formulation 7
1% DICLOFENAC IN 3't,_A Gel. 50 ml lar
Quantity 3000ml ~
~rm~l A -- ~u~pLier LOT Amount Per~en~ ~`
~-
Sterile Water Baxter AW45R6 2796ml -%
15 Glycerin BDH 2579 50g~71ml) 3%
Benzyl Alcohol BDH 23797 45g(43ml) 1.5%
Liquid wax DICDD Brooks 191-175 90 g 3%
Diclofenac Sodium Prosintex 9113003 30 g 1%
Hyaluronate Sodium Skymout HG 1004 90 g 3% .
20 MW 679,000
PROCEDURE
- Set up stirring apparatus using a 4 liter stainless steel
beaker.
- Add water, Glycerin, 3enzyl Alcohol and Liquid wax DICDD
25 and stir to mix thoroughly for 10 minutes ~;
Add Diclofenac Sodium and stir for 30 minutes to dissolve.
- Slowly add Hyaluronate Sodium, stirring at a high torque
initially during addition.
- After addition stir at a slower speed for 90 minutes.
- A white opaque viscous gel is formed.
-- ;;
2~ 2~ :
.. ~ ., :.
Formulation 8 ~ :
1% DICLOFENAC IN 3 . 0% HA Gel . 50 ml tube
Quantity 1500 ml
5 Formula Supplier LOT Amount Percent
Sterile Water Baxter AW45F1 1397 ml
Glycerin Life 1043 45g(36 ml) 3~ ~-
Benzyl Alcohol Caledon 02517 22.5g(22ml) 1.5%
10 Liquid wax DICDD Brooks 191-175 45 g 3%
Diclofenac Sodium Prosintex 9113003 15 g 1% : ~
Sodium Hyaluronate Skymart HG 1003 45 g 3% .
Mol. Wt. 661,600
P~CEQU~E
- Set up stirring apparatus using a 3 liter stainless steel
beaker. ~
- Add water, Glycerin, Benzyl Alcohol and Liquiwax DICDD, ~:
stir to mix for 10 minutes. i ...... ..... .
- Add Diclofenac Sodium and stir for 30 minutes to dissolve.
- Add Sodium Hyaluronate and stir for 90 minutes.
. . .. .
- 57 - 2 ~ $ 9 ~ 2 .L
`
Formulation 9
HYANALGESE CREAM (L)
50 ml tube ~ .
Quantity 3000 ml
5 FORMULA
A. Oily Phase SUPPLIER LO~ AMOUNT PERCENT
~lquid Wax DICDD 3rooks/Amisol 450g 15.0%
Br~kswax D ~rooks/Amisol 480g 16.0%
0 Glycerine Amisol 150g S.0%
A~ueous Phase
Sterile Water Baxter AW4YA8 1950ml -%
Meglumine Falk 150g 5.0% .
Sodium Hyaluronate Skymart PO1 45g 1.5% : -:
5 MW 207,000
Ibuprofen 8DH 150g 5.0~ . .
Suttôcide A Sutton 9,Og 0,3
~OCEDURE
A. - Add all the ingredients of the oily phase into a 4 liter
~tainless steel beaker, melt at 55C, finally heat to 75C - ~.
when aqueous phase is ready (at 75C) to pour in. :~
B. - Into another 4 liter stainless steel beaker, add 1950 ml
water. ~
- Set up the stirring apparatus and add the Meglumine
- Stir to dissolve with high torque, then slowly add
Ibuprofen ~ :.
- When the Ibuprofen is dissolved, slowly add Sodium
Hyaluronate
~ Stir cold for one hour to dissolve all the ingredients
- Finally heat to 75~C and stir thoroughly throughout a 30
minute period .:
- 58 ~ 2~ 2~
MIX B INTO A
- Slowly pour B into A (both at 75C) with stirring
- Immediately remove the hot plate (heat) and stir
- Stir with a strong vortex for one hour
- When the temperature is 45C, add the preservative
Suttocide A
- Stir for about an hour to cool to 35C
- At 35C remove the propeller and pour into 50 ml tubes
- Pour 50 grams of the cream into each tube
'
1~ BANAMINE IN 2.5''-i HA GEL
(L) XPB 041 Quantity 3000 ml .
FORMULA :
SUPPLIER LOT AMOUNT PE~CEN~
. :.
Sterile Water Boxter AW4SA2 2400 ml --%
Sodium Hyaluronite Skymart HE1003 75g 2.5% ~
MW 661,600 .
*Banamine, 100 ml vial Scheing O CNXB13 300 ml 1%
20 E~Da~e, 100 ml vial S~heir~Çr O CNX812 300 ml 1%
3000 ml
~50 mg/ml) 600 = 30,000mg ~
= 30 grams Flunixin in 600 ml
*Banamine contains Flunixin Meglumine (50 mg Flunixin per ml)
or 83 mg Flunixin Meglumine .
PROCEDURE
- Set up stirring apparatus using a 4 liter stainless
steel beaker
- Add water, stir with a strong vortex, then add sodium
Hyoluronate slowly
- Then immediately add the Banarnine, stir the mixture
for 4 hours.
One form of hyaluronic acid and/or salts thereof ~for
example sodium salt) and homologues, analogues, derivatives,
complexes, esters, fragments, and sub-units of hyaluronic acid,
~ 59 ~ 2~ 2~
.
preferably hyaluronic acid and salts and thereof, suitable for
use with Applicant's invention is a fraction supplied by Hyal
Pharmaceuticals Limited. One such fraction is a 15 ml vial of
Sodium hyaluronate 20mg/ml ~300mg/vial - Lot 2F3). The sodium
hyaluronate fractlon is a 2'~ solution with a mean average
molecular wei~ht of a~out 225,000. The fraction also contains
water q.s. which is triple ~is~illed and sterile in accordance
with the U.S.P. for injection formulations. The vials of
hyaluronic acid and/or salts thereof may be carried in a Type 1
borosilicate glass vial closed by a butyl stopper which does not
react with the contents of the vial.
The fraction of hyaluronic acid and/or salts thereof
(for example sodium salt) and homologues, analogues,
derivatives, complexes, esters, fragments, and sub-units of
hyaluronic acid, preferably hyaluronic acid and salts thereof,
may comprise hyaluronic acid and/or salts thereof having the
following characteristics:
a purified, substantially pyrogen-free fraction of
hyaluronic acid obtained from a natural source having at least
one characteristic selected from the group (and preferably all
characteristics) consisting of the following:
i) a molecular weight wlthin the range of
150,000-225,000;
.........
ii) less than about 1.25% sulphated mucopoly-
5 saccharides on a total weight basis;
iii) less than about 0.6% protein on a total
weight basis;
iv) less than about 150 ppm iron on a total
weight basis; -
.- ''' ''''',;
2 ~ 2 1.
v) less than about 15 ppm lead on a total
weight basisi ~;
vi) less than 0.0025% glucosamine;
vii) less than 0.025% glucuronic acid;
viii) less than 0.025'~, ~-acetylglucosamine;
ix) less than 0.0025~i amino acids; ~:
x) a ~V extinction coefficient at 257 nm of
less than about 0.275;
xi) a UV extinction coefficient at 280 nm of ~ ~-
less than about 0.25; and
xii) a pH within the range of 7.3-7.9.
Preferably, the hyaluronic acid is mixed with water and the ; .
fraction of hyaluronic acid has a mean average molecular weight
within the range of 150,000-225,000. More preferably, the
fraction of hyaluronic acid comprises at least one
characteristic selected from the group (and preferably all
characteristics) consisting of the following characteristics:
i) less than about l'ti sulphated
mucopolysaccharides on a total weight basis;
ii) less than about 0.49.i protein on a total
welght basis;
iii) less than about 100 ppm iron on a total
weight basis;
iv) less than about 10 ppm lead on a total
~ weight ~asis;
v) less than 0.00166% glucosamine;
vi) less than 0.0166% glucuronic acid;
vii) less than 0.0166"i ~-acetylglucosamine;
viii) less than 0.00166"i amino acids;
- 61 ~ 2 0 ~ 2 1
. ~
x) a ~V extinction coefficient at 257 nm of
less than about 0.23;
xi) a UV extinction coefficient at 280 nm of
less than 0.19; and
xii) a pH within the range of 7.5-7.7
Applicants also propose to use sodium hyaluronate -~
produced and supplied by LifeCorer~ Biomedical, Inc., having the ~;
following specifications~
Characteristics Specification
10 Appearance White to cream ~ ;
colored particles
Odor No perceptible odor
Viscosity Average < 750,000 Daltons -~
Molecular Weight
15 UV/Vis Scan, 190-820nm Matches reference scan
OD, 260nm < 0.25 OD units
Hyaluronidase Sensitivity Positive response :
IR Scan Matches reference
pH, 10mg/g solution 6.2 - 7.8
20 Water 8S`~ maximum ~ ;~
Protein < 0.3 mcg/mg NaHy
Acetate < 10.0 mcg/mg NaHy
Heavy Metals, maximum ppm
As Cd Cr Co Cu Fe Pb Hg Ni
25 2.0 5.0 5.0 10.0 10.0 25.0 10.0 10.0 5.0 ;
Microbial Bioburden None observed
Endotoxin < 0.07EU/mg NaHy
Biological Safety Testing Passes Rabbit Ocular
Toxicity Test
: '' '
- 62 - 2B3~621
Another form of sodium hyaluronate is sold under the
name Hyaluronan HA-M5070 by Skymart Enterprises, Inc, having the
following specifications:
Specifications' Test ~-
Results
Lot No. HG1004
pH 6.12 ..
Condroitin Sulfate not detected
Protein 0.05~,
Heavy Metals Not more than 20 ppm
Arsenic Not more than 2 ppm
Loss on Drying 2.07'',
Residue on Ignition 16.69.
Intrinsic Viscosity 12.75 dl/s (XW: 679,000)
Nitrogen 3.14~.,
Assay lOq.1'~;
Microbiological Counts 80/g
~. coli Negative
Mold and Yeast Not more than 50/g
- 63 -
2089~
Other forms of hyaluronic acid and/or its salts, and
homologues, derivatives, comple~es, esters, fragments and sub
units of hyaluronic acid may be chosen from other suppliers, for
example those described in prior art documents provided the form
of hyaluronic acid chosen is suitable for transport of the
medicine. ~ -
The following references teach hyaluronic acid,
sources thereof, and processes for the manufacture and recovery
thereof which may be suitable. ~ .
United States Patent q,141,973 teaches hyaluronic
acid fractions (including sodium salts) having~
"(a) an average molecular weight greater than
about 750,000, preferably greater than about
1,200,000 - that is, a limiting viscosity number
greater than about 1400 cm3/g., and preferably
greater than about 2000 cm3/g.; ;~
(b) a protein content of less than 0.5~ by
weight;
(c) ultraviolet light absorbance of a 1~3~ solution
of sodium hyaluronate of less than 3.0 at 257
nanometers wavelength and less than 2.0 at 280
nanometers wavelength;
(d) a kinematic viscosity of a 1% solution of ~ .
sodium hyaluronate in physiological buffer greater
than about 1000 centistokes, preferably greater
than 10,000 centistokes;
(e) a molar optical rotation of a 0.1 - 0.2~
sodium hyaluronate solution in physiological
2~8~21
. ~. .
buffer of less than -11 X 103 degree - cm2/mole
(of disaccharide) measured at 220 nanometersi
(f) no significant cellular infiltration of the
vitreous and anterior chamber, no flare in the
aqueous humour, no haze or flare in the vitreous,
and no pathological changes to the cornea, lens,
iris, retina, and choroid of the owl monkey eye
when one milliliter of a l~:i solution of sodium
hyaluronate dissolved in physiological bu~fer is
implanted in the vitreous replacing approximately
one-half the existing liquid vitreous, said HUA
being
(g) sterile and pyrogen free and
(h) non-antigenic."
15Canadian Letters Patent 1,205,031 (which refers to
United States Patent 4,141,973 as prior art) refers to
hyaluronic acid fractions having average molecular weights of
from 50,000 to 100,000; 250,000 to 350,000; and 500,000 to
730,000 and discusses processes of their manufac ure.
In order to determine the blood levels in patients
using formulations made according to embodiments of the
invention, a study of the pharmacokinetic profiles of two
topical diclofenac formulations after repeat dosing were
undertaken.
One such product was the product Voltarol Emulgel
marketed in the ~nited Kingdom by Geigy. The other was a
Diclofenac preparation in Hyaluronic Acid.
This was an open, repeat dose, crossover comparison
using a randomized balanced block in six healthy volunteers.
' ~ ' "'
- 65 - 2089b~:L
The study consisted of administration with one, two
week period in between periods, each period lasting fourteen
days. The test articles applied were for the first six days of ; ;
each period and the seventh day was study day during which the ~ :
final application is made and blood samples taken.
The approximate duration of the study including pre
and post study screening was six weeks.
Doses
Diclofenac (3.0%) with Hyaluronic Acid (2.5%)
Dose: Approximately 2 g, three times daily
Route: Topical ~
~Wl)Voltarol Emulgel, Diclofenac diethylammonium ~ ;
salt 1.16g aqueous gel (Geigy)
Dose: Approximately 2 g, three times daily ;~
Route: Topical (W1) ~
ADMINISTRATION~ suitablç patients ~:
Subjects applied one of the designated test articles
20 topically to the calves and massaged into the skin, in a dose of ~
approximately 2 g per application three times a day for six ;
consecutive days. The size of a 2g dose was prepared by
comparison with a silicone example given to each subject.
On the seventh day, the cream was appli.ed once, in the
same manner as before, under the supervision of the staff of the
Clinical Investigation Unit.
After a washout period of one week the procedure was
repeated with the alternate test article.
The following were the results of the tests~
(H = hyaluronic acid formulation)
(V = Voltarol Emulgel)
''': ~'
~0~21
PERIOD .l
All concentrations ng ml - 1
SUBJECT TIME POINT (hours)
0 0.25 0.5 1 2 3 4 5 6 8 10 12
H-1 10.3 7.1 6.4 ND ND 5.4 6.5 5.1 ND ND ND ND
H-2 ND 5.1 ND 5.1 ND ND ND ND ND 5.1 ND ND
ND ND ND 5.5 5.2 ND ND ND ND ND ND ND V-3
ND ND ND ND ND ND ND ND ND ND ND ND H- q
ND ND ND ND ND ND ND ND ND ND ~D ND V-5
N~ ND ND ND N~_ ND ND 8. q ND ND ND ND V-6
ND = NONE DETECTED (>5.0 ng ml~1)
e~ :, ,~
A11 concentrations ~g ml -1
SUBJECT TIME POINT (hours) -:
0 0.2.~ 0.5 1 2 3 4 5 6 8 10 12
V-1 ND ND ND ND ND ND ND ND ND ND ND ND
V-2 ND ND ND ND ND ND ND ND ND ND ND ND
H-3 ND ND ND ND ND ND ND ND ND ND ND ND
V-4 ND ND ND ND ND ND ND ND ND ND ND ND :~
H-5 ND ND ND ND ND ND ND ND ~D ND ND ND
1~- ~N~ NP ND ND ND N ~ ND ND ND ND ND ND
ND = NONE DETECTED (>5.0 ng ml~1)
Other tests were undertaken to determine blood levels :~:
comparing Proflex (a formulation containing Ibuprofen) and the
following formulation containing hyaluronic acid and Ibuprofen.
HYANALGESE CREAM (L) X PB 022
- 50 ml tube
Quantity 3000 ml
EQBI~l~ , ,"
,~,~ Oily Phase SUPPLIE~ LOT AMO[JNT pF.RCF.NT
Liquid Wax DICDD ' 3rooks/Amisol 450g 15.0%
Brookswax D E3rooks/Amisol 480g 16.0%
Glycerine Amisol 150g 5,0%
~ A~ueous Phase
Sterile Water aaxter AW4YA8 1950ml -% .
Meg1umine Falk 150g 5. 0% ~ : `
Sodium ~yaluronate Skymart PO1 4 5g 1. 5%
MW 207,000
Ibuprofen BDH 150g 5. 0
Suttocide A Sutton 9.0g 0.3
- 67 ~ 2 ~ 2
The following were the results
,.................... ................................... ............................ ... ~ ':
(A) PROFLEX ~ .
5 SUBJECT Time after administration (Hours) : :
Numher -~
~D O 0.25 0.5 l 2 3 4 5 6 8 10 1~
1ND 0.41 0.37 0.:37 v.32 v.3C '`.2/ '`.77 0.24 v.37 0.31 0.31 0.16
0 2 ND 0.12 0.1~ 0.08 0.1~ û.. ~ v.;~ i O.OR v.O9 0.08 ND 0,06
3ND 0.09 0.0~ 0.01 ND Nl~ NN Nli ND ND ND ND ND
4ND 0.12 O.lq 0.16 û,ll O.l' 3.25 O,~f~ 0.17 0.13 0.16 0.11 0.13 : -
5ND O.lq 0.19 0.19 0.15 0.36 v^.l6 ~ 0.12 0.11 û.~3 O.10 0.07 .
6 ND 0.11 0.09 0.09 0.06 O.Oi ()~o~ 0.0~, 0.05 NU N~ ND ND
Mean 0.0û 0.17 0.17 0.16 û.13 0.13 O.lf. 0.13 0.11 0.12 O.ll 0.09 0.07 ~.:
S.D. 0.ûû 0.12 0.10 0.11 0.10 0.10 0,lO O.lC 0.08 0.13 o.ll 0.12 0.06 ~ ~
(B) HYALURONIC ACID AND IBUPRO~EN .:~ :
SUBJECT Time after administration (Hours) . :.:.
Number
PD O Q.25 0.5 1 2 3 4 5 6 8 10 12
I ND 0.11 0.11 C.'.~ v.')R ~,.'`'i ;:.'`:.~ ;,.:: '`.'i '~.0l~ 0.11 0.16 0. l4
2 ND 0.22 0.7; 0.~.~, C,1l ~U~ 'U~ 3 ().L9 'J.l9 0.20 0.14
3 ND 0.1.7 D.~0 0.]~ ;).0~3 :";)~1 0.'1' '`.')(, N:, O.C~, D.26 0.09 0,05
~ ND ND Ni) Nl` Nli ~ N!~ N'i N:. NN ND ND ND
S ND ().11 ().;~j (,.16 1).`;' '`.v'3 ~ ) i";: ~".i` ~.09 ~.10 0.07 ND
6 Nr) 0,07 0.0/ ~,.0:~ NN `I!` N!) NN N,~ , ND ND Nr~
M~an 0.0û 0.17 0.11 0,1'5 'n~ .07 0.11 0.09 0.06S,D. 0.0û 0.0~ 1 ;).()6 ;,.~ J.~ ,,()'3 ~01 0.10 0.03 0.07
ND None detected <0.05 ,ug/ml
35The above clearly indicates that the blood levels are ~.;.;
much less using hyaluronic acid to administer the NSAID.
~ IXINBBy. REPORT
. .": .
A trial was conducted using a gel composition (Number
40109) comprising 3''~i Diclofenac in 2.5'~i Hylauronic Acid as
;previously described and a composition containing Diclofenac
sodium salt 3% but not including any form of hyaluronic acid.
(Number 112) The trial was conducted with 60 patients who were
randomly assigned to test preparations number 109 or 112. The
trial has not been completed as yet but so far 31 patients have
finished the protocol. Patients were diagnosed: ;
- 68 - ~a$~621
.
4 Rheumatoid arthritis of the knee
8 Myofascial trigger points in the M. trape~ius area
12 Periarthropathies of knee without effusion
7 Periarthropathles with effusion in the knee joint
~he 31 patients were aged 22-75 years (27 females, 4
males). All patients were hospitalized. Patients entering the
trial were thoroughly examined and type of extraarticular or
articular rheumatism assessed.
On day l baseline pain was assessed on the 10 cm
visual analogue scale (VAS) and pain measurement of the
quantititative pain sensitivity using a pressure tolerance meter
(PTM) were performed. Then test gel - appro.~imately 2.g - was
massaged on to the skin of ma~imum pain. Gels were applied 3
times daily. ,~
0.5, 1, 1.5, and 2 hours after morning application
measurements of pain sensitivity were carried out and values
recorded.
This procedure was countinued on day 2, 3 and 4;
measurements (VAS and PTM) of pain severity were done on day l, -
2 0 2 and 4 .
Prior of the beginning of the study and at the end on
day 4, physician's global assessment, assessment of swelling,
tenderness and limitation of movement were recorded. ;
,~ I ' ' '." ''', ';
' '
- 69 - 2
As the study is ongoing statistical evaluation is not .
yet available. For further details see Table 1. ; ~:
TABL~ 1 :
Composition Composition
~:
Reaction 109, n = 16 112, n - 15
Good Alleviation 13 8
Of pain
Moderate Alleviation 2 2 :::~
10 of pain
No Alleviation 1 5
Of pain . ~ ~
From the data recorded we have concluded that the - ;
patients to whom composition 109 was administered did better in
15 terms of earlier and longer lasting analgesic effect (up to 4 ~. ;
hours) than the 112 composition especially ln patients with
myofascial trigger points and with periarthropathies of the knee
~oints without effusions. Neither composition 109 nor
composition 112 treated patients showed any effect on swelling ; ::
if any swelling e~ist at all. Systemic side effects have not
been observed; one patient to whom composition 112 was
administered showed reddening of the skin on the site of
application.
Any intake of system NSAIDS,corticosteroids and ; ;
25 other analgesics was not allowed one week before and during the ;
trial. .
~ .
- 7- ~3~
``. . :
EXAMPLES
The following examples are offered to illustrate uses -~
of Applicants' invention. ;~
Example 1
A male patient had a number of lesions (basal cell
carcinoma), including one on his forehead which was a
combination of major "horny epithelium" and some degree of
ulceration. After continuous treatment with Formulation 1
(several times per day for several weeks of dosage amounts
squeezed from tubes as ribbons of composition), the lesions
.
showed epithelialization, no hemorrhagic areas, and no initiated
areas (as they were in the past without our treatmentj. The ~ ~ `
"horny epithelium" and ulceration of the forehead lesion were
also gone. The patient had a complete successful response with
the formulation. All basal cell carcinoma lesions had been
resolved and disappeared. There has been no recurrence.
'' ', :' '
Example 2 ;
60 year old male tennis player had sore elbow and
basal cell carcinoma on forearm proximate sore elbow. Patient
tried Formulation 1 to abate pain in tennis elbow. ~Dr. Falk
was not treating this patient for anything at the time, did not
know 'of the basal cell proximat-e the elbow and merely offered
the formulation for pain relief of the elbow instucting the
patient to squeeze a ribbon of the composition and apply and rub
into the sore elbow). However, the formulation "spilled" over
onto the Patient's basal cell carcinoma. Patient was planning
to have basal cell carcinoma removed surgically by another
:,
' '' ',,
~,`C~'~
- 71 ~ 2~
doctor, but when the patient returned to see the doctor, the
basal cell carcinoma was disappearing (because of spill-over of
Formulation 1). Dr. Falk was then advised and treatment was now
undertaken by Dr. Falk with direct application of Formulation 1
to the lesion 3 times a day for two additional weeks. After two
weeks, the basal cell carcinoma disappeared. There has been no
recurrence.
', ,
~xample 3
Male, mid to la~e 40's had severe basal cell carcinoma
on left temple. Doctors recommended its removal by surgery.
However, the surgery would have been risky because of the
lesion's proximity to facial nerves.
Patient saw Dr. Falk who gave him Formulation 2 to be
applied in dosage amounts 3 times daily.
After 19 days, 75O of the lesion was gone. Surgery
was postponed and the treatment was continued. Application of
dosage amounts of ~ormulation 2 was continued for an additional
two weeks. At the end of the 2- week period, the lesion was
completely resolved and disappeared without any surgery being
required. There has been no recurrence.
Exam~le 4
`~ Male, early 40's, had recurrent actinic keratoses
lesion on his right temple. Early attempts at removal by third
parties involved the application of liquid nitrogen (twice)
without final resolution. The lesion kept recurring. The
patient was sent to Dr. Falk who treated the lesion with
Formulation 1 with applications of dosage amounts 3 times daily
~; ~
j~ 2~$~
for 7 days. After 7 days, the lesion was completely resolved
with no subsequent recurrence.
Example 5 ~;
A male patient suffering from kyphosis suffered from
constant back pain. Taking analgesics orally and rubbing back
preparations onto his back did little to alleviate the back ~;
pain. When NSAIDs in hyaluronic acid (sodium hyaluronate) were
applied directly to the back, the back pain eased and
10 disappeared. ;
With indomethacin (dissolved in N-methyl glucamine)
and naproxen both dissolved in hyaluronic acid, the patient .
experienced some side effects. However, with Toradol~M (the ~:
~+/-] form tromethamine salt of ketorolac - a prostaglandin
biosynthesis inhibitor and analgesic and anti-inflammatory, the
back pain eased and disappeared for some time and there were no
side effects. The compositions were applied generously onto the
sites of back pain.
','.:"..,: :.
~xample 6
A male patient with basal cell carci.noma was first
treated by an oncologist who attempted to surgically excise the
lesion (without success) and then irradiated the lesion again
without success. The patient then attended before Dr. Falk who ;
applied Applicant's formulation (diclofenac with sodium
hyaluronate and excipients). Application was made three times
daily for about a month and the lesion disappeared. Some
excoriation anterio~ and slightly superior developed over the
',. ' '" , ',
'~'.':'.'`''"';;,~'''
~ 73 ~ 2~ 21
. ~ .
last two weeks but was cleare~ by the application of hyaluronic
acid by itself. - ~-
This resolution clearly indicates that even with prior
applications of unsuccessful therapies (surgery and
irradiation), Applicant's formulations can be used successfully.
Example 7
In another patient, a drug (methotrexate) was carried
in hyaluronic acid and applied topically to a patient with
psoriasis. The formulation was absorbed and the psoriasis
cleared. ~ -
Example 8
A patient with dermal (skin) metastases in a fibratic
15 scar form and metastatlc cancer in the form of musculoskeletal ~ ~-
involvement in her thora.Y.
On copical application of our formulation comprising
diclofenac (Voltaren) in hyaluronic acid (sodium hyaluronate),
her pain decreased dramatically and her skin and boney
20 involvements steadily improved. ~
,. " ,::"
TOPICAL DICLOFENAC ACID 3li IN HYALVRONIC
ACID GEL (2.5'~i) BASE
A practitioner reviewed the effectiveness of topical
Diclofenac Acid 3% in hyaluronic acid gel (2.5%) base in acute
traumatic injuries of no longer than 3 days duration. The cases
were all in the spectrum of ages between 18 and 65. Normal
exclusion criteria were followed regarding exclusion of
~ 74 ~ 2 a(~
,
pregnancy, aspirin or N.S.A.I.D., allergies or active peptic
ulceration.
As an overall, the following lmpressions were gained
from 30 cases: :
1. The topical H.D.(composition comprising sodium
hyaluronate and diclofenac) had an obvious analgesic
action with onset occurring rapidly within one hour;
this is a phenomenon not obviously seen with other
non-steroidals that we have used.
2. There was a very definite patient acceptance of the
gel as a form of treatment, being logical, easy to
apply, without local or systemic side effects, rapid
absorption with no staining of clothing.
3. The anti-inflammatory action was equivalent on a
"yuestimate" based on experience of similar
injuries to oral N.S.A.I.D.s, without the threat or ~
risk of side effects. ;
In summary, compared with other topical N.S.A.I.D.s
the analgesic effect is distinct, the anti-inflammatory is equal
to oral N.S.A.I.D.s and the patients' acceptance is far superior
to any other diclofenac or piroxicam topical that the
practitioner evaluated.
Following the practitioner's basic preamble regardlng ;~
the parallelism of topical N.S.A.I.D.s and topical steroids,the :
practitioner has used the former in contact dermatitis, insect
bites and U.V. erothema, all with very positive effects, again
pointing direction to trials of a double blind nature in these
fields.
',~ ':;"'
,:- .,'' :' ''
- 75 - 2~ 21
Positive (P)
Patients Date Negative (N)
Inltials Of Fil ~nable to
(M~ nr (F~ p~irth I~o. n;~qnos~ Comrnents on Outcom~ Comment (U~ ;
0 LA (M) 11.04.56 ~yper- Severe discomfort following P
aesthesla extensive surgery to dorsal
spine with insertion of Example of
rods in l9~9. Rven contact peripheral
with clothes produced sig- action on
nificant discomfort. super- ~ `
Initially treated with EMLA sensitiza- -
with only transient anaes- tion of
thetic results, however nerve
even after 3 days treatment ending
with Hyal diclofenac acid queried.
noticed marked decrease in
supersensitivity which has -
continued for at least 9 - -
weeks while still using gel. ~
KB ~l 08.06.58 Chronic Treated right knee which was P
chondro- worse initially and was
malacia amazed at the response, then
perhaps started to treat left knee :
dating back that was not so painful,
to 19-6. again with positive response.
Here we have a built-in con-
trol.
35 o~ ~F~ Chronlc Inltially felt some improve- N
neurogenic me m which was not continued
pain in although initially quite
ankle with positive - query placebo
associated reaction.
dysaesthesia
DC ~F) 07.11.51 ChronLc back N
pain - query
due to ~acet
4 5 9yndrome or
trigqer
points,
nosLs
unceFtain.
_ _ _ Positive (P)
~atients Date Neqative (N)
Initials Of Flla Unable to
~M~ or f~ Rlrth No. n~aonoels Comme~ts on OlltcomQ Comment ~U
CC 18.01.25 Chronic cap- Definite effect over knee P
sulitis where application to tar-
right hip get distance short. No
right knee obvious effect over hip. ~-~
AG (F) 07.11.58 Myositis in Initially given placebo in P
rhomboids error, only marginal or
muscles minimal effect, if any.
following Found active to be effect-
motor vehicl tive while being used, did
accident not cure condition which
needed trigger point
therapy.
CH ~F) 22.00.61 Chronic No significant effect, nor N
relapsing has aggressive therapy since
tendonitis including lnjection with
right elbow cortisone and numerous ~ ;
opinions.
SH ~F) 16.07.55 Tendonitis Control of tendonitis while
and myositis using preparation. Is now
back at work.
DM ~) 17.06,47 Neuronitis This patient has a very
unusual pain in his left -
3S groin following nerve
in~ury, with the use of
preparation noticed de- -
crease in pain sensatlon
while on medication,
Hyperaesthesia altered ~ i
although pain (whlch may be
. phantom) still present.
:': '." :~i~ ,'
PJ 15,06.45 Capsulitis Symptoms lmproved 50~ while U of right using Hyal diclofenac acid,
wrlst however, on discontir.uation
pain reappeared, Exact
etiology uncertaln.
_ _ ' ~;
, ~ '" :'
:, ` ~ ' :' ':. .. '
., :. ' .
~ ' .,
: ~ ~ ' ,.,:
,
. .
- 77 - 2~
~,~C,~ 3% ~ICLOFENAC ACID (Hll)
Positlve ~P)
5 Patients Date Negative (Nl
Initials Of Fil Unable to
~M) cr ~ R~rth ~ Dla~nos~s Comments on Outcome Comment ~U)
DJ ~F) Dorsal Control while using gel
0 myosltis equal and with less side ~ ~-
effects than tiger balm.
Controlled symptoms while
using medication. Exact
diagnosis as to cause of
myositis uncertain.
: :
DK ~F) 27.08.38 Severe This patient has had capsu- P
capsu1itis litis left shoulder for many
left shoulde years and treated wlth only Extremely
transient relief with corti- rewarding
sone in~ections, poor relief case
with topical piroxicam. Was
started on topical diclo-
fenac acid and notlced
relief of pain in 20 minutes
continuinq for 4 - 6 hours.
See letter March 1l/92. At
present is using H.D. regu-
larly, has found it to be
useful in other areas of
chronic paln. Is President
North Amerlcan Chronic.
PaLn Assoclation, has good
insight into medication and
placebos etc. Has two D.C.S.
_ implan~s.
', ~ .
,
: , :
:: :
8 2 ~ 2 ~
~IRONIt: CON~ITI~NS - EVAL;IA~115
_ Posltive ~p)
5 Patients Date Negative IN)
Initials Of Fil Unable to
IM~ or (~) rth No. r)l~gnoc~s CommPnts Qn_Qutssm~_ _ Cnmment ~U~
JL ~M~ 10.12.45 Chronic Pain has failed to respond Nmyositis to many aggressive treat-
secondary to ments.
query facet
syndrome
RMC~F) 13.06.57 Neuronitis It is a difficult case with U
following considerable overlay, she
facet obtained some relief with
rhizotomy H.D., would estimate 30-40
with result- Interestingly hyper-
ing pain in anaesthesia was decreased. ~ -
her back -
RM IF) 20.08.52 Chronic Using H.D. significant
capsulitis improvement in pain while
used, on stopping treatment
recurrence of pain, needed
intra-articular cortisone. : ~ ;
GM ~F) Sub-acute Rapid resolution of pain p
tendonitis within one day and positive
right ankle return of function.
PM ~F) 20.09.46 Acute on Rapid analgesic response P
chronic with rapid settlement.
3 5 OSteo-
arthritis of :
flrst meta-
tarsal
phalyngeal . ' ',
9 0 ioints . ~ ~.
DN ~F) 10.03.44 Chronlc Excellent response to appli- P fascialtls catLon of H.D. with occlu-
of feet sion, Had failed to respond : ~
4 5 to oral N.S.A.I. D . s and . : .
physiotherapy. Query posi-
tive result due to sh~rt
application target distance
in a vascular tissue.
50 , . . . - ~;
' ' ~
: '::
"
.
:,
:' ' .: ''
. . ~ ~ , .
: ~
::
_ 79 _ ~ i2:~
, ~ .
EIYAI.URQ~LC Al:~ID ~CTH 3% DICLOEE2~IC ACI~189
Posltive ~P~ ~
5 Patients Date Negative (N) ~-
Initlals Of Fil Unable to
~Ml or (Fl B;rrh _~L_ n;a9nos;s Comments on Outcome Comment ~)
~P (F) 09.03.20 Severe Initially one knee treated P
chronic with such good results that
arthritis of both knees treated, see Side
the knee, letter. Not only did pain effects-
Snable to decrease but marked swell- non/Inci-
take oral ing around knees. Signifi- dental
U.S.A.I.D.s cant relief of pain and resolutlon
increase in movement as a of area of
result of this and perhaps thrombo-
reduction of swelling, phlebitis
Tntrestinqly has severe below area
R~rlPrf i Ci al v~o~P vPin~. of treat
~eve1ODed thromboDh1e~L~is ment
aro~lnd r;ght knPP ~nd the
area treated by chance
showed ~ar less redness and
2 5 t~L~ n~ss than the throm-
hnphl Pb; t;~ bP1ow t h;s are~.
SP (M) 06.11.48 Idio- Has had similar episodes U
pathic with poor response to many
diffuse treatments including
capsulltis N,S,A,I,D,s per os
of hands
WS 04.06.45 Chronic Has been exposed to number-
neuronitis ous ereatments including
due to tow attempts of surgery
injury to without effect. There is
lateral decrease in hyperaesthesia
cutaneous but no change in pain.
nerve of
thigh
MS ~F) 04.06.28 Chronic Failed to respond to number P
capsulitis of treatments, good back-
ground resolution of pain,
however, still had acute
pain with certain movements.
,
~ '~
i`
,
'
' .
: - .
~, '. ..
'
- 80 - 23~95~1
,
. . . .
eositive ~P) -
Patients Date Negative (N)
Initials Of Fil Unable to ~ -
(~ or ~ ~irth No, Diaanosis Comments on outcQme- Comment (Ul
IS ~F~ 15.01.48 Chronic Had failed to respond to
capsulitis numerous treatments -~
including oral and topical
N,S.A.I.D.s Using H.D
there was equivalent control
of pain as with other
therapies which lasted while
medication was used.
Referred for surgical
opinion.
G~ ~F) 26.03.q7 Chronic Oral diclofenac acid dis- P
tendo- continued due to gastritis
sinovitis and also history of ulcera-
tion. tion. Control using H.D.
equal to or better than oral
N.S.A.I.D.s.
VK ~F) 01.01.39 Chronic Good relief of pain and
tendonitis tenderness whiLe using H.D. for pain
however on discontinuation N
of gel symptoms returned, for
treated with intramuscular resolution
steroids.
G~ (~) 03.11.21 Acute on In view of age and general
chronic parous medical condition, Commented
osteo- ideal for topical. Had on better
arthritls been previously on topical absorption
left hand piroxicam for left shoulder compared
capsulitis. to topical ' '
piro~icam
- '' '''' `
. .
.
,
, ~
, ~
'' ~ `:'.
' : "
81 2 ~ 2 :1
. . .
_~ .
Positive IP)
5 Patients Date Negative (N)
Initials Of Fil Unable to
IM~ or (F~ R;rth _ NQ_ ~;agn~Lia Comme~ts on Qi~s~e C~mment IU)
JA ~M) 06.02.58 Severe post- Produced good superficial P
eraumatic analgesia especially where
and surgical staples were irritating sub-
osteo- cutaneous tissue, little
arthritls of effect on deeper, severs
left leg osteoarthritic pain of knee.
with staples This pain was of consider-
Poor result able severity, needing nar-
to oral cotics.
N.S.A.I.D.s
also gastric
2 0 irritation.
IM (M) 30.11.51 Chronic Severe rhomboid inflammation P
superficlal right side, treated with
myositis H.D., very definite improve-
ment in pain and tenderness.
~K ~F) 23.04.70 Acute on Excellent rapid analgesic P
chronic followed by anti-inflammator
capsulitis response in young women who
due to could not take oral N.S.A.D.
sports due to past gastritis.
in~ury right
hand
AD ~F) 03.01.49 Chronic Poor response to H.D. After N
diffuse pain intensive investigation and
thought to numerous consultations and
be myositls treatment, pain still un-
diagnosed and unresponsive,
NH ~F) 25.03.25 Subacute Excellent response analgesic P
capsulitis and anti-inflammatory wise
right ankle within a few days. Marked
clinical improvement. In
view of this patient's
parous general medical con-
dition and hypertension, not
suitable for oral NSAIDs.
_ _
.:
;
- 82 - 2~
_~ -
Positlve ~P)
5 Patients Date ~egatlve (N)
Initlals Of Fil ~nable to
IM~ or ~F~ ~1 rth o. n1a~Qsi~L comments on Qu~ssn~L___ comm~nt ~IT~
MD 18.04.34 Subacute Had failed to respond to
rheumatoid oral N.S.~.I.D.s, which
arthritls caused gastritis, tried on
topical piroxicam with neqa-
tive effects. Negative
response to H.D.
15 MW (F) 07.05.46 Heberden's Very slow positive outcome, P
nodes, pain- initially improvement in
ful, swollen pain followed by reduction
causing in swelling. Etiology of
difficulty this condition is unknown,
20 in movement partly genetic. Would have
been interesting to treat ~
alternate digits, plus or ~-
minus thermographic confir- -.
matlon. :
LP ~F) 20.07.23 Acute on Initially treated with
sub-acute Idarac, poor response over- ~:
osteo- all. some improvement in
arthritis of generalised arthritis of ~
the hands hands but none on Heberden's -
with nodes. Pain flared on stop-
Heberden's piny Idarac due to gastritis.
nodes Started on H.D... especially
favourable results with sub-
5idence of tenderness of
nodes and settling of
arthritis. Interestingly
enauqh, no flare up on dis-
continuation after one month ~ :
40 _ _
"~ '.
', .,
~:', . ' :' ~`
:~
~ ! ,
: ~ ' " '',',':
'~ ' . ,,~
'
: ''' ~ . ' ' :.
, ~ ~ .''',
- 83 - 2~
ay~Lh~NIc ~C~ L~ c~ as~-Ac3lL ~HD?
S Positive (P)
Patlents Dat~ Neqative ~N)
Initials Of Fil Unable to
~M~ or !F) R1 rth No. ~acnos15 ~ Co~m~nt (U\ _
0 JG ~F) 24~11.50 Post facet Had failed to respond to U
rhlzotomy oral ~.S.A.I.D.s and
hyper- E.M.L.A. Application of
aesthesia, H.D. improved the surface
wlth marked pain significantly but had
pain and no effect on the deeper
hyper- pain. My impression ~as
aesthesia that the deeper pain was
between due to section of the facet
scapulae nerve and beyond the reach
of the t,opical medication.
There is llttle doubt that
the skin sensitivity was
decreased,
sw ~F) 10.09.39 Etnee pain Upset in past due to oral P
due to N.S.A.T,D.s., also hypor- ~Effec-
. chondro tension made one loathe to ti~e while
malacia use this medication with being
serum L~vels. Good anal- ~sed)
gesic and anti-inElammatory Condition
acSion, however on di~con- only cured
tinuation pain flared. Seen by surgery
for arthroscopic surgery
wlth relief of pain.
* Two types of pain-response in only one
1. Interestlngly in the whole series, there was not one
case of local side effects and as expected from past studies, no
general or systemic. Since this report was prepared we have had
one case of mild folliculitis which responded to discontinuation
of treatment, will rechallenge.
2. A number of patients commented that tney felt the gel
lmproved the texture and softness of their skin, and commented
` that it was messy`or stained their clothes.
3. In one case of topical thrombophlebitis where the
inflamed vein crossed the area of treatment, the vein in the
.
area of treatment improved while that outside at a distance did
not. Again, slmilar to using oral N.S.A.I.D.s. ***.
.
.,
:,
- 84 -
2Q$~
Photographs were ta~en of patients with basal cell
carcinoma Figures 1-6 photographs, and of mice with tumors
induced in the skin of the hind legs (Figure 7 photographs).
The patients were treated by using combinations of NSAIDS, (non~
steroidal anti-inflammatory drugs) and hyaluronic acid
(including sodium hyaluronate) according to the invention (3% -
diclofenac in 2.5'~ sodium hyaluronate gel base). Each of the six
sets of Figures made up of photographs of the different persons
include a legend describing or explaining each picture
appropriately labelled. The mice had tumours induced in the
skin of their hind legs and dosage amounts (2ml) of Novatrone
(10 mg. per dosage amount) (MITOXANTRONE (t.m.) and 2.5% sodium
hyaluronate were applied (rubbed onto) the skin at the site of
the pathology. The tumours reduced in si~e clearly illustrating
the percutaneous delivery of the medicine by the hyaluronic
acid. (See Figure 7). ;
The following additional comments are made with
respect to the patients.
With respect to R.W. and Figure 5, the reader will
note in Figures Sa and 5c the patient suffered from basal cell
carcinoma on his back (Figure 5c) and his temple (Figure 5a).
Because of the age of the individual (86) the basal cell
carcinoma on his back could not be reached by him for
application of the medication. Thus the basal cell carcinoma in
5c remained untreated and grew (see Figure 5d). However, the
portion indicated in 5a on his temple could-be reached and after
application of the basal cell carcinoma formulation to the
temple and forehead the results are as in 5b; the basal cell
",'.
- 85 - 2 ~8 9 ~1
carcinoma is disappearing. Thus, the gentlemen's own method of
treatment acted as a control.
With respect to R.F. and Figures 4, two areas of basal
cell carcinoma in need of treatment are shown by the arrows in
Figures 4a and 4c and the results are shown in Figures 4b and 4d
as indicated by the arrows after treatment with Applicant's
invention.
With respect to H.A., male, and Figures 3, Figure 3a
indicates two areas of basal cell carcinoma by the arrows,
close-ups of which are shown in Figures 3c and 3e. After
treatment with the NSAIDS and HA gel three times a day for the
period between January 26, 1992 and March 16, 1992 the basal
cell carcinoma is clearing as per Figures 3b, 3d and 3f.
The same is true with respect to male M.F. and Figures
2 which appears clear in the photographs (see Figure 2a and the
response shown in Figure 2b).
With respect to male, W.D. and Figures 1, the upper
, : ..,
lesion in Figure la (indicated by the upper arrow) is gone after
treatment with Applicant's invention (See Figure lb) and the two
lower lesions in Figure la are well on their way to disappearing
(See Figure lb).
With respect to female D and Figure 6, the lesion was
left untreated for a long period and gradually encompassed her
eye. Surgery could not be undertaken without jeopardiz1ng the
eye. ~y applying Applicant's invention (dosage amounts) over a
prolonged period, the basal cell carcinoma has constantly
decreased in size.
With respect to Figure 7, (7a) shows mice having
tumors in the skin induced in their hind legs. After continuous
- 86 - 2~
applications to the shaved hind legs having the tumors in the
skin by rubbing in dosage amounts by Applicant's invention, the
tumors have decreased in si~e.
The effect of Hyaluronic acid as a drug carrier of
anti-cancer agent (5-FU) 5-Fluoracil was also studied.
(Intratumour injection study)
B. EXP~IMENTAL ~ODEL (2) ~-~
1. Method and Material
a. Animal : Fisher 344 rat, male
200-250g
b. Tumor model
Fisher Bladder Carcinoma
Tumor (2mm viable tumor fragment) was
transplanted subcutaneously on the right
frank by trocar
c. Treatment was started when t~mor size is about
l.S cm.
(2 weeks after implantation.) .
1. These drugs were administrated by intratumor in~ection.
~right frank)
At the same time, injection into normal skin (left frank)
was carried out similarly.
Group A : H-5-FU Smg/kg + saline /0.3ml ~i.t.)
B : H-S-FU 5m~/kg ~ HA 15 mg/kg /0.3ml (s.c.)
3H-FU without or with HA was injected as a single dose
(0.3ml) into the center of the tumor (on the right frank) with a
30 gause needle. At the same time, injection into normal skin
~on the left frank) was carried out similarly.
The tumor and skin was then removed at different times
(lh,6hr) for counting radioactivity of the remaining content in
the tissue.
2. Result
All the results were expressed as Mean + S.E.
- 87 -
2 ~ 2 ~
TVA~IOR TISSU~ NO~blAL SKIN
5cpm
10000 ¢9~ 10~0
5-FU ~Agroup ~ 5-FI~ ~A yr~up
~ 5~FUgnoup ~4~ ~ ~-FUgroup ~ 4)
8000 p~ 000
~ ~:
6000 ~ I ~ 6~ ~
~i I ~ ~
15 ~ I ~ ~ I~L
1hr 6hf lhr ~hr
3. Conclusion
1. In S-FU_HA aroup radioactivity was accumulated
and retained in the tumor tissue for a long period, whereas
rapid clearance was demonstrated in normal tissue. (skin)
2. In S-FU group, radioactivity immediately
disappeared from the tumor or the normal tissue
by diffusion, primarily into blood capillaries.
---- SFU can traverse freely between the interstitial
space and blood capillary.
- a8 - 2~39~;r~l
The Effect of Hyaluronic Acid as a Drug Carrier in Target Cancer
Chemotherapy
A. E~PERIMENTAL MOD~ Intravenou~ Injection)
1 . ~ethod and Mat~rial
a. Animal : Fisher 344 rat, male
200-250g
b. Tumor model
Fisher Bladder Carcinoma
Tumor (2mm viable tumor fragment) was transplanted
subcutaneously on the right frank by trocar
c Treatment was started when tumor size is about 1.5 cm.
15 ~2 weeks after implantation.)... tumor weight:1.0 + 0.3g ~ ~
The drug was administered Intravenously (through the ~ -
penile vein)
Group A : 5-FU 20mg/kg (3H-5-FU30~Ci) + saline
B : 5-FU 20mg/kg (3H-5-FU30~Ci) + HA 15mg/kg
C : 5-FU 20mg/kg ~3H-5-FU30~Ci) + HA 15mg/kg
+ (3H-HA30~Ci)
2 . Sampl e Col l ection .
a. accumulation of ADR, 5-FU in tumor tissue and liver
(1). Tumor was surgically removed (and blood was
collected) at *predeterminated time after drug
administration. Tumor weight was measured (and
blood was centrifuged to obtain a plasma ;
sample.)
* lSmin, 60 min, 3hr, 4hrs,.... after drug
administration
Liver was removed for radioactivity counting at
the same time.
(2). Radioactivity level in tumor tissue was
counted, using a liquid scintillation counter.
.. .. .
- 89 ~ 2 ~ ~ 9 ~2
3. Conclusion
Radioactivity in Tumor Tissue and ~ive~
- T~mor _ Liver
15min 3H-5FU (n=6) 2810+165 18680+625
S 3H-5FU+HA (n=6) 352+190 23593il460
3H-5FV+3H-HA (n=4) 4087~681 32060+2145
60min 3H-5FU (n=3) 1751+149 5451+841 ;
3H-5FU+HA (n=4) 2599+489 8265+1849
3hrs 3H-5FU (n=6) 1493+227 2230+449
3H-5FU+HA (n=6) 2512+449 2897+340
3H-5FU+3H-HA (n=4) 3606+929 6977+1633
lS Shrs 3H-FU (n=3) 853+129 1129+70
3H-5FU+HA (n=3) 1981+479 1754+248 ~ .
_ 3H-5FU+3H-HA (n=3) 2L68+163 3018+325
mean+ S.E. ~.
HA : 15 mg/kg (30~Ci/kg) .
Rwbu~hnm~nb~
~X 5-FU : 20mg/kg (30~Ci/kg)
25 4400L_ --_ ~U 1. Radioactivity in tumor tisaue in 5-FU
+ HA group is higher than that in 5-FU
group. There is ~ignificant difference
\ (p>0.05, ANOVA) between with and without
~o~ \ \ HA at 3hrs after injection. The high
~ ~ intratumor concentration was retained
\lr~ ~ for a prolonged time in S-FU+HA group.
\ll ~ ~Thiq retention waq confirmed by the
~0 ~ ~C' intratumor injection study.)
~ ~ 2. These results teach that HA can
~ ; enhance S-FU uptake in tumor tissue.
1~ _ This phenomenon reaults from HA
, distribution (in tumor tis~ue HA may be
lost from the extracellular matrix) and
O ~ , , the va~cular uniqueneqs of tumor tissue
40 ~ 4~ ~dDu~4 ~hyperpermiability of tumor veqsel~ to
macromolecular drug, HA).
As many changes can be made to the invention without
departing from the scope of the invention, it is intended that ~
all material contained herein be interpreted as illustrative of .
the invention and not in a limiting sense.
