Note: Descriptions are shown in the official language in which they were submitted.
`` 116~39LC~3
BACKGROUND OF THE INVENTION
Field of the Invention
The present invention relates to a new method
for extracting propolis from clean raw propolis as well as un-
processed beeswax to obtain a dry propolis powder. In addition,
the invention includes a uniaue water soluble dry propolis
powder. Propol~s is a resinous compound collected by honey
bees from various plants and the buds of different trees.
Propolis is also known as ''honey bee glue" and ~s used by the
bees to coat parts of the interior of the hi~ve and to seal
the cracks and crevices of the h;`ve.
Description of the Pri~or Art
The existence of propolis, or honey bee glue,
has of course long been recognized by apiarists. In fact, it
has been variously described as both a ~less~ng and a curse.
While the substance has~been largely~lgnored as a necessary,
sticky evil associated with beekeepina, some rather surprising
: ~ -
studies o$ propolis have been made with partlcular regard to
comm~rcial and therapeutic uses for the material. Most of the
20 ~ research~has ~een conducted in Asian and European countries,
~- but potential commercial uses for propoli~s have been acXnow-
ledged in the United States.
In a relatively brief article authored by Dr.
F.B. Wells and published ~n the Novemberj 1976 i`ssue of the
~- American Bee Journal at pa~es 512, 513 and 542, the potential
for use of propolis and propolis-containing preparat;~ons for
therapeutic purposes is outlined. As is indicated in that
;~ article, and the foot notes to the art~cle, the majority of
research on propolis has been undertaken in Great Britain,
r ' . .,
.
L6~Q;~
Denmark, Poland, Russia, Romania, Czechoslovakia, East Germany,
Yugoslavia and Bulgaria. These reports are in general agreement
that raw propolis consists essentially of approximately 55~
resins and balsams, 30~ wax, 10~ etheral oils and 5% pollen.
However, it will be appreciated that the composition of propolis
may vary and is dependent upon its geographic area and the
season of its collection. Nevertheless, reported laboratory
and clinical tests are quite consistent in their observations
- that propolis and propolis-containing compositions do exhibit
bactericidal effects. It has been suggested that propolis may
be responsible for the relatively low concentration of bacteria
and molds in the atmosphere within bee hives. As early as 1965
three Romanian investigators reported that alcoholic extracts
of propolis, drones and royal jelly had virulicidal activity
against Type A influenza virus in vitro. Russian Patent No.
267,014 claims the eff;~cacy of~an alcoholic extract of propolis
in combination with glycerin for treating conjunctivitis.
Russian Patent No~ 232,470 discloses and claims an alcoholic
,
extract of propolis as part of a toothpaste composition possess-
~20 ~ing both prophylactic and antiseptic properties.~ Romania
Patent No. 48,101 relates to a cosmetic lotion including an
alcoholic extract of propoli$ also including boric acid. British
Patent No. 1,465,194 teaches a method comprisin~ repetitive
freezing and thawing of propolis to obtain a material suitable
for subsequent therapeuti~c uses.
Thus, one must clearly admit that the preparation
of alcoholic extracts containing propolis is well known and,
further, that organic solvents are normally used to prepare the
extracts. In fact, as the literature clearly teaches, ethyl
alcohol might be termed the universally-accepted solvent. On
: ~ .
-- 2
.
~L~68~ 3
.
the other hand, one must also admit that the literature is
quite lacking in that it fails to teach controlled, reproducible
methods for extracting propolis of known constituent composition.
There is virtually no teaching in the prior art of any means
for obtaining a water soluble propolis extract, and the litera-
ture repea~edly refers to propolis as being substantially
insoluble.
It is therefore apparent that there is a great
need in the art for a method of obtaining a substantially pure
propolis extract of known and reproducible constituent composi-
tion. Furthermore, due to the recognized potentlal for using
propolis extracts for therapeutic purposes, it would be
extremely desirable if a water solu~le extract could be obtained.
It is therefore a purpose of this invent;on to provide a new
extraction and purification-process which~is reproducible to
o~tain a dry propolis powder of known and relatively constant
composition. A further o~ect of the present invention resides
:
in a method ~cr obtalning-a water soluble dry propolis powder
~20: which has heretofore-not ~een reported.
SUMM~RY OF THE INVENTION
T e present inventlon relates~ to a method for
extracting propolis as well as for purifying the propolis
extract to obtain a dry propoIis powder suitable for a variety
of uses including, but not necessarily limited to! cosmetic
and therapeutic application~ The invention further relates to
a method for obtaining a water soluble dry propolis powder and
the water soluble propolis powder itself. Accordingly, the
present invention comprises a method for extracting the propolis
using both organic solvents, aqueous solutions of organic
solvents, and mixtures thereof. It is to be understood that
- 3 -
~ -
: .
' '' ' ' ~ :
the particular solvent utilized in the extraction method will
be chosen with regard to the intended end use of the dry pro-
polis powder and whether or not it is desired that the powder
be water soluble or organic solvent soluble~
: The procedure for preparing the dry propolis
powder is first to obtain a quantity of raw material consisting
essentially of either clean raw propolis or unprocessed bees-
wax. The raw material is preferably placed in an amber glass
container and covered with solvent. If clean raw propolis is
used as the raw material, about 2 liter.s of solvent are added
~ per kilogram or propolis. If unprocessed beeswax is used as
: ~ the raw material, about 3 liters of solvent are used per kilo-
gram of beeswax.
~:~ As wlll be set forth in:greater detail herein-
after, preferred solvents for use in the method of this inven-
~:~ tion are absolute ethanol .(eth.~l alcohol~ and aqueous solutions
:: of ethyl alcohol. However, laboratory experimentation has
revealed that the following organic solvents, as well as
~ aqueous solutions of these solvents, may ~e used: ~ :
:~20~ : isopropyl alcohol
n-butyl alcohol-
sec-butyl alcohol
. tert-butyl alcohol
. ~ ethyl ether
enzyl alcohol
;~ propylene glycol
dimethyl sulfoxide
ethylene glycol
n-propyl alcohol
;. ~ethyl alcohol
benzyl benzoate
acetone
polyethylene glycol and
~.~i
~ 4 -.
t " . .
.,
:~68~
glacial acetic acid.
It is of course to be understood that the
solvent utilized should be of high quality and purity, consis-
tent with the final uses of the dry propolis powder.
While the initial extraction preferably takes
place at room temperature, acceptable results are given at
temperatures from about 0 C to about 37 C. The extractin~
vessel containing the raw material and solvent is shaken several
times daily for a period of from one to ten daysl preferably
at least three days. At the end of this time, the extract is
filtered through Whatman No. 1 filter paperl or its equivalent.
At this point it should be noted that the once extracted raw
material may be again covered with solvent and re-extracted as
descrlbed above to obtain additional propolis-contalning
filtrates~
The solvent may then be removed from the propolis-
containing filtrate to yield a dry propolis powder. Removal
of the solvent may be accomplished by lyophilization or incuba- -
-::
tlon~(evaporation~. Depending upon the solvent used in carry-
20 ~ ing out the extraction method, the resulting dry~propolis
powder will be either water soluble or organic solvent soluble.
~ .
- For example~ if ethyl alcohol is used as the
solvent, an or~anlc solvent soluble propolis powder will be
obtained. However, if a 10%-25% aqueous ethyl alcohol solution
is used as the solvent, water soluble dry propolis powder will
be obtained.
This invention may further comprise a method
for purifying the propolis extract prior to obtaining the final
dry propolis powder. Purification comprises cooling the
30i~ propolls-containing filtrate to a temperature of at most about
- 5 -
.
. ~
'
34~3
:`
-20 C for approximately 24 hours. At the end of this period
the extract will become viscous and opaque~ While maintaining
the filtrate at about -20 C it is shaken and filtered through
Whatman No. 50 filter paper, or its equivalent. During the
cold filtration procedure, which is carried out at about -20 C,
a very clear filtrate should be obtained, and the waxy material
remaining on the filter paper contains waste materials. If
the filtrate is not clear, the procedure of cold filtration
must be repeated.
The clear filtrate resulting from the cold
filtration process may be filtered at the reduced temperature
or may be brought to room temperature and filtered through a
0~2 micron filtration system. The solvent is then removed
from this final filtrate as by lyophilization or incubation to
obtain the purified dry propolis powder. In light of the rela-
tively low temperatures utilized in the purification method
outlined above, it is to be understood that this purification
.ethod is not suitable for aqueous solvent solutions, since
the water would freeze.
~20 ~ While the above procedure has consistently proved
to~yield an extremely pure dry propolis powder of consistent
composition, further quality control over the final powdered
' .
product may be obtained in the following fashion. After the
cold filtration process descxibed above, but prior to removing
the solvent, the cold process filtrate may be brought to an
extremely low temperature such as, for example, -70 C. If,
after about 24 hours at that temperature, the filtrate is still
clear, purity of the final product is assured~ Any cloudiness
- 6 -
mab/,~
. . .
''
.
in such a sample would indicate the unacceptability of the
filtrate for further processing.
Detailed examples of various extraction and
purification me-thods in accord with this invention are pre-
sented below. Also as will be set forth in greater detail
hereinafter, the resulting dry propolis powder may be utilized
as a constituent in many cosmetic and therapeutic substances.
The invention accordingly comprises the several
steps and the relation of one or more of such steps with respect
~ to each of the others, and the composition possessing the
features, properties and the relation of constituents which are
exemplified in the followinc detailed description, and the
scope of the invention will be indicated in the cIaims.
~ DETAILED DESCRIPTION
.. . I
The following examples are set-forth ln order to
fully describe the method for extracting and purlfying propolis
.
as well as the resultlng dry propolls powder and its uses.
EXAMPLE I
- About 500 grams of clean raw propolis was placed
.
~20 in an amher glass container and covered with about 1 liter of
absolute ethanol~ This mixture was allowed to sit for ten days
at room temperature with periodic agitation several times each
day. At the end of ten days, the mixture was filtered through
.
Whatman No. 1~ filter paper. The resulting propolis-containing
filtrate was then incubated at about 70 C until a dry propolis
powder was obtained. Incubation temperatures as low as about
55 C may also be employed, but greater time to obtain the final
dry product will be required. Care should be taken, however,
with regard to any increase of the drying temperature, for the
- ? -
:
.
` :~ 16t~
propolis will burn at about 80 C.
The dry material remaining after incubation is thedry propolis powder, and is organic solvent soluble.
EXAMPLE II
The method of Example I was repeated;-however,
the solvent was removed from the propolis-containing filtrate
by lyophilization (freeze drying) after partial reduction of the
alcohol content by evaporation. The final product obtained by
this method was not a powder, but a "gummy" propolis residue.
- EXAMPLE III
The method of Example II was repeated utilizing a
15~ aqueous ethanol solution as the solvent. This resulted in
a dry propolis powder which was water soluble. Chemical analysis
of the water soluble dry propolis powder gave the following
results per 100 grams of dry propoiis powder:
Calcium 0.33 grams
,
Phosphorous 0.111 grams
Albumin 3.7 grams
Protein 18.5 grams
. ,~ .
Creatinine ~ 118.5 milligrams
Bilirubin 55.5 milligrams
; ~ Glucose 26.1 grams
.
Alkaline Phosphatase 4,148 International Units
- Potassium 0.397 grams
Sodium 0.085 grams
;~ Zinc 0.299 milligrams
Vitamin B12 (estimation) 0.133 milligrams
; Folic Acid (estimation) 1.926 milligrams.
Furthermore, study resulting from application of the resulting
:;.
khfll~Ç
~' :
1~6~ 3
water soluble propolis to smooth muscle tissue revealed that
this propolis contained no antihistamine properties quite un-
like most drugs used today for treating virus symptoms. The
presence of creatinine, bilirubin and alkaline phosphatase in
the dry propolis powder is quite remarkable and may provide
the basis for other uses of the powder, since these are
normally found in animal tissue.
EXAMPLE IV
The method of Example III was repeated utilizing
a 10~ aqueous ethanol solution as the solvent. The results of
this procedure were substantially identical to those of Example
III.
EXAMPLE V
~he method of Example III was repeated utilizing
a 20% aqueous ethanol solution as the solvent. The results of
- this procedure were substantially identical to those of Example
III.
EX~LE VI
,
The~method of Example III was repeated utilizing
a 25% aqueous ethanol solution as the solvent. The results of
,
this procedure were substantially identical to those of Example
III.
EXAMPLE VII
The method of Example III was repeated utilizing
~ a 30~ aqueous ethanol solution as the solvent. The propolis-
- containing filtrate derived according to this method was deemed
unsuitable because of excessive cloudiness. It is therefore
believed that water soluble dry propolis powder may only be
obtained using aqueous solvent solutions of no more than about
_ 9 _
- :
.
~' ~
:' ,
.
,
25% concentration.
EXAMPLE VI I I
About 500 grams of clean raw propolis were
placed in an amber glass container and covered with about 1
liter of absolute ethanol. The mixture was allowed to sit at
room temperature for about ten days with shaking several times
each day. At the end of the ten days the extract was filtered
through Whatman No. 1 filter paper.
To further purify the propolis-containing
filtrate, it was cooled to a temperature of about -20 C for
about 24 hours. At the end of this period it was observed that
the extract had become viscous and opaque. The chilled filtrate
was then shaken and filtered while maintaining the temperature
at about -20 C, through Whatman No. 50 filter paper. This
resulted in a very clear purified propolis-containing filtrate.
The solvent was removed from the purified filtrate by incubation
at about 70'' C for approximately 48 hours, or until dry. The
resulting material was organic solvent soîuble dry propolis
, ,
powder.
20 ~ EXAMPLE IX
Utilizing about 100 grams unprocessed beeswax
as the raw material and about 300 ml ethanol as the solvent, the
method of Example VIII was repeated and yielded-similar results.
Of course, a smaller quantlty of purified dry propolis powder
was obtained.
EXAMPLE X
~ The method of Example VIII was repeated; however,
; the solvent was removed from the purified filtrate by
lyophilization after partial reduction as in Example II.
'.';
- 1 0 -
` : :
.
A3
.
Chemical analysis of the final product obtained according to
this procedure yielded results substantially identical to those
reported above in Example II.
EXAMPLE XI
The method of Example VIII was repeated utilizing
isopropyl alcohol as the solvent. Equivalent results were
obtained.
EXAMPLE XII
The method of Example VIII was repeated utilizing
n-butyl alcohol as the solvent. The results of this procedure
were substantially identical to those of Example VIII.
.
EXAMPLE XIII
The method of Example VIII was repeated utilizing
sec-butyl alcohol as the solvent. The results of this procedure
were substantially-identical to those of Example VIII.
EXAMPLE XIV
The method of Example VIII was repeated utilizing
ter~-but~,~l alcohol as the s~lvent~ The results of this pro-
~ :
cedure were substantially identical to those of Example VIII.
.
EX~MPLE XV
he method of Example VIII was repeated utilizing
~ ~ ethyl ether as the solvent. The results of this procedure were
; ~ substantially identical to those of Example VIII.
EX~IPLE XVI
The method of Example VIII was repeated utilizing
benzyl alcohol as the solvent. The results of this procedure
:` :
were substantially identical to those of Example VIII.
EXAMPLE XVII
The method of Example VIII was repeated utilizing
. .
.
':
- , . .
'
. ~ . . .
G~4~3
propylene glycol as the solvent. The results of this pro-
cedure were substantially identical to those of Example VIII.
EXAMPLE XVIII
The method of Example VIII was repeated utilizing
dimethyl sulfoxide as the solvent. The results of this pro-
cedure were substantially identical to those of Example VIII.
EXAMPLE XIX
The method of Example VIII was repeated utilizing
ethylene glycol as the solvent. The results of this procedure
were substantially identical to those of Example VIII.
EXAMPLE XX
The method of Example VIII was repeated utilizlng
n-propyl alcohol as the solvent. The results of this procedure
were substantially identical to those of Example VIII.
EXAMPLE XXI `
The method of Example VIII was repeated utilizing
methyl alcohol as the solvent. The results of this procedure
were substantially identical to those of Example VIII.
~ ~ EXAMPLE XXII
The method of Example VIII was repeated utilizing
benzyl benzoate as the solvent. The results of this procedure
were substantially identical to those of Example VIII.
EXAMPLE XXIII
The method of Example VIII was repeated utilizing
acetone as the solvent. The results of this procedure were
; ~ substantially identical to those of Example VIII.
` ~ ~ EXAMPLE XXIV
The method of Example VIII was repeated utilizing
polyethyIene glycol as the solvent. The results of this
- 12 -
kh~
.,
~3. 68~3
procedure were substantially identical to those of Example
VIII.
EXAMPLE XXV
The method of Example I was repeated utilizing
glacial acetic acid as the solvent. The results of this
procedure were substantially identical to those of Example I.
EXAMPLE XXVI
The method of Example VIII was repeated utilizing
a mixture of equal parts of ethyl alcohol and methyl alcohol as
the solvent. The results of this procedure were substantially
identical to those of Example VIII.
EXAMPLE XXVII
The following procedures were conducted in order
to determine the bactericidal activity of the purified propolis
powder obtained in accord with the procedures set forth above
in Example VIII.
A 10%, by weight, dry propolis powder solution
in absolute ethanol was prepared. This 10~ solution was then
.
used to prepare a variety of additional solutions, diluted with
~20 ~distilled water, to give final concentrations Gf dry propolis
powder of from less than 10 milligrams to 10 milligrams of
propolis powder per milliliter of solution. These solutions
were then again diluted with a microbiological culture medium
such as supplemented peptone broth II obtained from The Becton-
Dickinson Corporation of Rutherford, New Jersy, ~.S.A. The
final solutions with peptone broth II varied from less than 1
to 10 milligrams of propolis per milliliter of peptone broth II
solution. After about 24 - 48 hours incubation at about 37 C
with each of the organisms listed below, the cultures were then
- 13 -
.
. . . .
.
~6~33
replanted on the blood agar plates prior to a second incubation
at about 37 C for about 24 hours. The three organisms utilized
were present at a level of 15 million per 1 milliliter and were:
Staphylococcus aureus ATCC 25923
Escherichia coli ATCC 35933
Pseudomonas aeruglnosa ATCC 27~53.
With the organisms tested individually in con-
trolled studies, the propolis was found to have the following
effects. Purified propolis powder in the final concentration
of 2 milligrams per milliliter is lethal to Staphylococcus
aureus. Purified propolis powder in a final concentration of
6 milligrams per milliliter is lethal to Escherichia coli.
Purified propolis powder in a final concentration of 5 milli-
grams per milliliter is lethal to Pseudomonas aeruginosa.
Similar experimentation was performed utillzing
water soluble propolis powder obtained in accord with the
method of Example IV. The bactericidal activity of this
propolis was equivalent to that just stated above.
- EXAMPLE XXVIII
~ The following procedures were followed in order
to determine the antioxidative properties of water soluble dry
propolis powder obtained in accord with the procedures of
Example IV~ 250 micrograms of the water soluble dry propolis
powder were dissolved in 1 ml of distilled water. To this was
- added 0~02 ml of 0.1 N potassium permanganateO Decolorization
took place approximateIy 2.1 seconds giving a positive
indic~tion of the antioxidative properties of the water soluble
dry propolis powder. These results are deemed important for
the reason that the antioxidative properties of the propolis
.
.
- ! 14
~.
: - " ' '.~
powder correlate to its bactericidal efficacy.
Dry propolis powder ob~ained in accord with
the method of this invention is considered as a storage
material available for immediate use. It is believed that
the stability of the propolis powder, when stored in an amber
glass container at room temperature, is at least 10 years.
The propolis powder is soluble in all the solvents mentioned
in the methods given in the above examples including, obviously,
water. Furthermore, organic propolis solutions are also soluble
in glycerol and many chemical surfactants used in the pharma-
ceutical, cosmetic and food industries. Similarly, organic
solutions of propolis powder are readily miscible with castor
oil. The incorporation of organic solutions of propolis powder
and different vegetable oils can be achieved fLrst t~rough the
combination of the organic solution with castor oil or with
chemical surfactants. Mineral oil, after combination with
vegetable oil, creates suitable conditions for the addition of
organic ~ropolis solutions. Similar results can be a~hieved
through the combination of organic propolis solutions with
20~ ~ ~ chemical surfactants and then with mineral oil. ~
Use of the dry propolis powder in organic or
aqueous solutions, creams, lotions, suppositories, douches or
,
in other pharmaceutical or cosmetic bases possess significant
antibacterial, antifungal and antiviral acitivity. The
observed anti~iral activity of the dry propolis powder is
particularly signlficant, and has been observed to be effective
against Herpes simplex (type 1), Herpes simple~ (type 2), Zoster
; virus, Epstein ~arr virus and common cold viruses. Infectious
hepatitis and distemper have not as yet been investigated, but
: .
- 15 -
;, .. . .
. .
1~6&~3
.
theoretical considerations strongly indicate the efficacy of
the extracted dry propolis powder.
In relatively large concentrations propolis
extracted according to the method of this invention possesses
anesthetic properties. Large concentrations also enhance
animal tissue metabolism and may be applied locally to increase
blood circulation. The increase in tissue metabolism during
the treatment indicates that the stimulated tissue may react
against inflammatory processes and that the propolis possesses
regenerative properties. Increased blood circulation in local
tissue is an important factor in comhatting cellulites, cramps
and other conditions. As will be indicated below, the oral
application of 5 to 10% propolis powder in absolute ethanol
has proved to be effective for bacterial, fungal and viral
diseases. ~
Of particular note is the efficacy of such
solutions in the treatment of Herpes simplex (types 1 and 2).
~erp~s simplex (type 1) is spread by contact of the mucous
~ membranes which results in cold sores. Herpes simplex (type 2)
infections are spread by sexual contact, and are considered as
the most common venereal disease in the United States. It is
generally accepted that Herpes simpIex (type 2) is linked with
cervical cancer. Both of the Herpes viruses can cause
encephalitis which has a high mortality rate. Propolis powder
prepared in accord with the method of this invention and
applied in the form of creams or ointments rapidly cures cold
sores. Herpes simplex (type 2) infections of the sex organs
can be cured by a specially applied douche. In both cases,
- su~sequent oral treatment is recommended in order to prevent
- 16 -
: ~d!.`~,
kh
.
" ' '' ' ~ ' ` ~ ' '
~,
1~84~
recurrence of the diseases.
In vitro studies of Herpes simplex (types 1
and 2) have demonstrated that about 10 micrograms of the
propolis powder per milliliter of the culture media kills the
viruses without affecting cell division. Oral administration
in dosages o about 2 to 3 grams per day will prevent further
outbreaks.
It should also be noted that the dry propolis
powder obtained in accord with the method of this invention
has been observed to be extremely efficacious against the
majority of common cold viruses when treated immediately after
the first symptons of the cold appear (no later than 2 or 3
hours after the onset of the condition). When the propolis is
taken more than about 3 hours after symptoms appear, the severity
of the symptoms do appear to diminish.
The following examples, then, are given for the
purpose of illustrating various formulations for topical and
oral administration of the dry propolis powder.
EXAMPLE XXIX
~20 Petrolatum Propolis Ointment
A 0.5 to 5.0~ propolis-ethanol solution ma~ be
incorporated into white or yellow petrolatum in concentrations
up to 60%, by volume, propolis solution. To decrease the
viscosity of this and other ointments, mineral oil, wool fats,
animal fats or fish fats can be used.
- EXAMPLE XXX
Spermaceti Cream
The following ingredients are mixed to obtain the -
cream with all composition constituents listed in weight
- 17 -
33
percents:
Polyoxyethylene 20 sorbitan monostearate 8.0
Propolis powder 2.0~
Sorbitan monos.earate 8.0%
Spermaceti 10.0%
Water q.s. ad 100.0%.
EXAMPLE XXXI
Anti-Herpes Cream 1
The following formulation, with consituents
listed in ~eight percents, has proved to be effective in
treating Herpes viruses:
- Propolis powder 1.0 - 5.0%
- Dermabase q.s. ad 100.0%.
Dermabase is a hypo-allergenic cream base which
.:
~ is compatible with most medicaments. Its pH is close to that
: .
of human skin. Dermabase is a vehicle for topical application
used~in pharmaceutical, cosmetic and veterinary preparations.
It~is manufdctured by Professional Pharmaceutical Corporation,
2795 Bates Road~ Montreal, Quebecl H3S:lB6,;~Canada.
~ - EXAMPLE XXXII
Anti-Herpes Cream 2
The following formulation with constituents
listed in weight percents,~has proved to~be effective in
treating Herpes viruses:
Propolis powder 0.5 - 10.0
.
Unibase q.s~ ad 100.0~. ~
Unibase is a dermatological all-purpose base,
and~has a pH approximating that of the skin. Unibase is
manufactured by Parke-Davis & Company, Ltd., Box 633, Station
:
~ - 18 -
- -: - ~ -:., . ,i
- , ~ .
:
: :.
''' ' ' '
"A", Scarborough, Ontario, MlK 5C5, Canada.
EXAMPLE XXXIII
Suppositories
The propolis powder prepared in accord with the
method of this invention may be administered in suppositories
prepared in accord with the following formula wherein all
constituents are listed in weight percents:
Polyoxyethylene 20 sorbitan :.
monostearate 35.0-55.0%
Polyoxyethylene 4 sorbitan
monostearate 40.0~60.0%
Propolis powder l.0- 5.0%.
EXAMPLE XXXIV
Suppositories
; Yet another formulation for suppositories in-
cluding powdered propolis may be prepared as follows, again :_
with all constituents listed in weight percents~
Glyceryl laurate 6.0-16.0%
Polyoxyethylene 4 sorbitan
~:~ 20 monostearate - . 82.0-92.0%
Propolis powder ~ 2.0%.
: : . EXAMPLE XXXV
Oral Preparations
: ~ A liquid (syrup) preparation for the oral
.
: administration of propolis powder may be prepared according to
the following formula:
Polyoxyethylene 20 sorbitan
monooleate 60 grams
50% Propolis ethanol solution 30 grams
-- 1 g -- -
Propylene glycol 100 grams
Ethyl alcohol 100 grams
70% aqueous sorbitol
solution, USP 300 grams
Distilled water 410 grams
flavoring agents and/or preservatives may be included as
desired.
The following examples are presented for the
purpose of setting forth propolis-containing solutions which
have proved to be efficacious for treating the common cold.
EXAMPLE XXXVI
10 milliliters (approximately 1 tablespoon) of
about 10% propolis-ethanol solution are mixed in a glass (about
8 ounces) of water or juice. Alternatively, the propolis-
,
ethanol solution may be mixed with coffee or tea giving the
appearance of added milk.
- This mixture may be taken about 3 times a day, but
if first administered immediately alter the first symptoms are
noted, the symptoms may disappear within a few hours.
EXAMPLE XXXVII
Nasal Ointment for Cold or Hay Fever
; Petrolatum 96.0 grams
50% Propolis-ethanol solution 4.0 grams.
This ointment should be applied to the nostrils a
few times daily as required. Other similar bases may be sub-
stituted for the petrolatum.
EX~MPLE XXXVIII
EIoney-Propolis Formula
Honey 80.0-90.0 grams
- 20 -
kh~i" `.
:;
.
~6~ 3
- 25~ Propolis in concentrated propylene glycol,
USP 10.0-20.0 grams.
EXAMPLE XXXIX
Gelatin Capsules
Sorbitol solution, VSP 80.0-90.0 grams
25% Propolis in concentrated propylene glycol,
USP 10.0-20.0 grams.
The above is only one example of many possible
means of incorporating propolis into capsules. The propylene
glycol USP potentiates the activity of alcoholic propolis
solutions up to 6.5 fold with respect to cidal activities versus
numerous microorganisms.
I~ has also been determined that propolis obtained
in accord with the methods of this invention is useful in the
treatment of respiratory tract infections and inflammatory
processes of the lungs, includiny bronchial asthma, sinusitis
~nd hay fever. One means of treatment is inhalation therapy in
which the drug is dissolved in hot water, and the vapors are
inhaled according to conventional procedures. The following
example sets forth a formula and procedure for the inhalation of
propolis.
EXAMPLE XL
About 10 milliliters (approximately 1 tablespoon)
of 10% alcoholic solution of propolis is added to about 1 liter
of hot water and mixed well. The vapors are inhaled.
EXAMPLE XLI
Yet another formulation for inhalation therapy
may be prepared as follows:
50% propolis-ethanol solution 10 ~rams
.~ ,'63 - 21 -
~L~61 3~G~3
Gum Benzoin 8 grams
Storax 6 grams
Tolu balsam 2 grams
Aloe 2 grams.
The above is blended into 100 milliliters of
g0% ethanol. In a preferred use, 1 teaspoon of this mixture
is added to 500 milliliters of hot water to produce the
inhalation solution.
Other solvents of propolis for inhalation therapy
include benzyl alcohol and polyethylene glycol.
In studies of asthma, in experimental animals it
has been determined that polyethylene glycol may be used as a
solvent for propolis in the formulation of injectable prepara-
tions (intramuscular).
As indicated above in Example XXXIX, propolis in
gelatin capsules with suitable non-ionic surfactants such as,
for example, polyoxyethylene 20 sorbitan monostearate, can be
used in the treatment of gastroirltestinal tract inections,
ulcers and other inflammatory disorders of the bowel. Treatment
2~ can be acheived with the oral application of propolis in con-
~unction with nonionic surfactants and propylene glycol in the
form of gelatin capsules. Yet another treatment form comprises
the use of retention enemas including propolis.
The gelatin capsules should be formulated to
maximize the absorption of propolis in the gastrointestinal
tract. This can be achieved by mixing the propolis-ethanol
solution with sorbitol or diluted propylene glycol solution
until a milky appearance of the combined substances results.
This will avoid or minimize coating of the gastric mucosa.
- 22 -
. -- . .
The daily intake of propolis should range
within about 3-6 grams. The retention enemas shou~d be
given in quantities of about 200 milliliters three times a
week or according to a physician's directions. The pH o~
the enema solution should be between 5.0 and 6Ø The
concentration of propolis may vary from about 5 to about 10
percent.
It has also been determined that the propolis
can be mixed with concentrated propylene glycol ~SP and in-
corporated in the gelatin capsules which, when ingested, can
coat the stomach walls. The capsules in contact with gastric
juice will form a milky suspension which adsorbs over the
surface of the stomach cells forming a thin film. This coating
has been shown to suppress the appetite for 2 to 3 hours, and
such treatment is recommended for the control of weight in
obese patients.
Vaginitis, cervicitis and cervical erosions may
be treated with 1 to 5~ propolis-eth nol solution in combination
with glycerine, propylene glycol and with or without non-ionic
surfactants in the form of douches, suppositories or ointments.
- Propolis powder can be used in otolaryngology and
renal infections in the form of solutions, tablets or capsules.
Propolis powder can be incorporated into ointments
for the treatment of burns.
For cystic flbrosis the daily oral application
of one gram of propolis powder should be administered in the
form of capsules or solutions.
Patients with ieukocyte dysfunction disorders
often develop recurrent bacterial infections which cannot be
- 23 -
controlled by current methods of therapy. Propolis powder
solutions, being strong bactericidal agents, can be used
orally in the treatment of these disorders. The oral applica-
tion of propolis powder solutions in a daily dose of 2 to 3
grams calculated on an anhydrous basis for a period of ten
days is recommended. They can also be used in conjunction with
current antibiotic therapy.
The regenerative and rejuvenating properties of
propolis powder can be used in areas of medical science such
as plastic surgery and dental surgery. Propolis ~owder in the
form of ointments, creams, lotions, solutions, shampoos, cream
rinses, shaving lotions and scalp creams in contact with the
damaged or diseased lesion on the animal or human body shows
considerahle healina and rejuvenatin~ properties. These
regenerative propeXties of propolis powder can be observed in
dental sur~ery, plastic surgery of animal organs and also in
vitro tissue culture studies.
When proplis powder solutions are added to a fibro-
blast cell culture, the number of mitotic cells increases. Such
studies demonstrate an increase in the enzymes responsible for
,
- the increased metabolism within the cell. The increased enzyme
actlvity occurs with the following enzymes:
Adenosine triphosphatase
Acid phosphatase
Glucose-6-phosphatase
Succinate dehydrogenase
This may explain why propolis powder solutions show regenerative
properties such as the increase in the ac ivity of formation of
a collagen.
- 24 -
Propolis powder may be consistently employed
in conjunction with other antibacterial agents used in
dentistry. Propolis powder incorporated in suitable bases
will control surperficial and deep infections of the mucous
membranes and bone, and will disinfect tooth cavities or root
canals.
Propolis powder solutions used alone or in
combination with benzocaine is an excellent anaesthetic. An
illustrative formula follows.
EXAMPLE XLII
Propolis Benzocaine Solution
Ethyl aminobenæoate 7.5 grams
Propolis powder in propylene glycol to make 150 milliliters.
The final concentration of propolis in the above
formula may vary from 1.0 to 10.0%, by weight.
Propolis powder may be used in the form of
liquids or pastes to relieve post extraction pain (alveolar
analgesic). An example of a paste fol1Ows:
EXAMPLE XLIII
Propolis Benzocaine Paste
Lanolin alcohols10.0 grams
~ellow beeswax10.0 grams
Petrolatum10.0 grams
Ethyl aminobenzoate2.0 grams
Clove oil 3.0 grams
50~ Propolis ethanol solution 15.0 grams.
Propolis in the above formula ac-ts as anti-
microbial, analgesic, anaesthetic and regenerative agents.
It speeds re~eneration of the tissues and reduces inflammatory
.. ..
~.~684g~3
infiltrations.
EXAMPLE XLIV
Propolis in a Mouth Rinse Solution
2-10% Propolis in alcohol-glycerol solution
40.0%, by volume
Propylene ~lycol 10.0%, by volume
Distilled water 49.8%, by volume
Flavoring agent 0.2%, by volume
In this formula sorbitol USP or non-ionic
surfactants may be used. Propolis powder has wide ranging
applications in the treatment of dermatological disorders.
Propolis powder for this purpose can be incorporated into
ointments, creams, lotions, solutions, shampoos, cream rinses,
- douche and oral preparations. Propolis powder can also be
incorporated into currently used pharmaceutical and cosmetic
products directly or with suitable surfactants.
- Ringworm may be treated by the local application
of 10~ propolis-aicohol solution or 5 to 10% propolis powder in
~ an ointment made of petrolatum or another base two to three
times daily for four weeks, or until symptoms are no longer
evident.
Psoriasis, seborrheic dermatitis, eczema and neurodermatitis
may be successfully treated with from 0.5 to 25% propolis powder
in the form of lotions, solutions or ointments prepared with
bases as previously described.
Propolis powder in concentrations from 0.5 to
10.0% may be formulated with wool fat or its alcohols and in
combination with petrolatum and mineral oil for treating corns,
warts and calluses.
- 26 -
Propolis concentrations of from 0.25 to 1.0%
combined with non-ionic surfactants and incorporated into
shampoos, cream rinses or creams are efficacious for treating
dandruff. Similarly, propolis solutions in concentrations
varying from 0.25 to 2.0% may be incorporated into hypo-
allergenic bases for treating poison ivy and jellyfish
dermatitis.
Acne vulgaris may be treated with up to 3%
propolis solutions combined with up to 3% salicylic acid,
non-ionic surfactants such as polyoxyethylene 20 sorbitan
monolaurate and with 40% ethyl alcohol or another suitable
alcohol. Other compounds can also be used such as methyl
salicylate, glycerine, or propylene glycol.
- A most effective treatment for hematomas and
other brulses consists of about 2-5% propolis-ethanol solution
made into an ointment with petrolatum. The ointment is spread
on the bandage, which is then taped over the affected area.
The unique p~operties of propolis ~lave been
used to create a new range of cosmetics. Propolis powders and
solutions are endowed with preservative and antioxidant
properties and natural skin rejuvenating properties which are
indispensible to the many cosmetic preparations~ The con-
centration of propolis in its powdered form can vary from 12ss
than 1% to greater than 2%. As a preservative or an anti-
bacterial and antifungal agent propolis solutions may be used
in concentrations varying from 1.0 to 2.0% on an anhydrous
basis. In antiwrinkle preparations the percentage of propolis
powder can be increased. The rejuvenating and other properties
of propolis in trials of a number of cosmetic formulas have
- 27 -
fulfilled expectations.
Propolis powders may be used in moisturizers,
night and day creams, nutrient creams, barrier creams, cuticle
creams, cleansing creams, lotions, cold creams, mask prepara-
tions, all types o~ lotions and solutions, shampoos, con-
ditioners, cream rinses, shaving lotions, finger nail polish,
soaps, lipStickc, baby creams, baby lotions, anti-diaper rash
products, message creams, massage lotions, ski~ rubbing pro-
ducts, aerosols for cosmetic or medical purposes, keratolytic
(desquamating) products and anti-cellulite products.
Moisturizing creams are used during the day
under makeup or as a foundation cream endowing the skin with a
soft appearance, retaining its moisture and preventing wrinkles.
An acceptable formulation for such a moisturizing cream follows:
EXAMPLE XLV
Cream base 78.0 grams
Propylene glycol10.0 grams
Avocado oil 10.0 grams
50% Propolis solution2.0 grams
Propylene glycol may be substituted with glycerol, sorbitol
solution USP or lanolin. Avocado oil my be substituted with
another vegetable oilj or an animal or fish fat.
EXAMPLE XLVI
~old creams
These contain large amounts of fatty or oil
ingredients. An example follows:
Cream base 40.0 grams
Oil or fat 58.0 grams
50% Propolis solution2.0 grams
- 2~ -
. ~ , ,. ~ . .
EXA~iPLE XLVII
Night Cream
Cream base 70.0 grams
Oil or fat 27.0 grams
50% Propolis solution3.0 grams
The increased quantity of propolis is an important
factor in the rejuvenation of the skin during the night when
the mitotic activity of the skin is increased. Humectants
may be added to aid in retaining skin moisture.
EXAMPLE XLVIII
Cuticle Cream
This softens the cuticles and prevents the nails
from becoming brittle. It actually toughens and thickens the
nails permitting them to be grown longer. A formula follows:
Petrolatum-lanolin base 97.0 grams
50% Propolis solution 3.0 grams
EXAMPLE XLIX
Facial Masks
Facial masks are intended as skin cleansing and
tlghtening agents. The presence of propolis aids in rejuvenating
the skin. The following is one example of a facial mask:
- Fuller's earth 50.0~, by volume
50% Glycerol solution 44.0%, by volume
50% Propolis solution 5.7%, by volume
Perfume, if desired 0.3%, by volume
China clay, kaolin or bentonite may also be used in combinations
with or as a replacement of the Fuller's earth used above.
Propolis powder may also be used in liniments
in con~unction with oil of turpentine, capsicum, extracts or
9 _
kh/~
.
.
4~
arnica, linseed oil, camphor and isopropyl alcohol, etc.
As previously indicated, propolis powder and
propolis solutions may be used as preservatives, anti-oxldants,
stabilizers and rejuvenating agents in a variety of current
cosmetic products. Their incorporation would require only
slight modifications of the original formulas. The propolis
may be directly incorporated or via the use of a solvent or
surfactant. The surfactants can be non-ionic, anionic or
cationic.
Propolis powder and solutions may also be used
in the food industry as preservatives, anti-oxidants and
stabilizers of food products, with emphasis on preserving
animal and fish fats. In the distilling industries they can be
incorporated into alcohol ~or use in oral treatment of patients.
In the tobacco industry it is believed they can be used as a
flavor or as a medicated agent in anti-ashmatic cigarettes.
It should also be noted that the following
solvents, because of their chemical equivalency to the solvents
previously described, could also be used in the method of this
invention:
Amyl alcohol
.
Isoamyl alcohol
Phenetyl alcohol
Ethylene glycol ethyl ether
Isoamyl benzoate
Isoamyl butyrate
Isoamyl formate
Isoamyl isovalerate
- 30 -
kh/~
Isoamyl salicylate
Formamide.
In similar fashion, it is anticipated that other, equivalent
organic compounds which are solvents for resins and balsams
may be used in the method of this invention.
Briefly summarizing, then, it can be seen that
the present invention presents a unique method for preparing
dry propolis powders which are suitable for a wide variety of
end use applications. It is to be emphasized that the method
and resulting water soluble propolis powder is deemed quite
significant for the reason that heretofore water soluble forms
of propolis were not known.
It will thus be seen that the objects set forth
above, among those made apparent from the preceeding discrip-
tion, are efficiently attained and since certain cha~ges may
be made in carrying out the above method and in the composition
set forth without departing from the scope of the invention, it
is intended that all ~.atte~ contained iI' the above descriptior
shall be interpreted as illustrative and not in a limiting
sense.
It is also to be understood that the following
- claims are intended to cover all of the generic and specific
features of the invention herein described, and all statements
of the scope of the inventions which, as a matter of languageO
might be said to fall therebetween.
kh/~
