Note: Descriptions are shown in the official language in which they were submitted.
~l~774~7
BACKGROUND OF THE NVENTION
This invention relates to the composition and
preparation of an antimicrobial agent which may be used in
dental preparations, surgical and other soaps, various other
topical preparations, injectable medicines, and other drug
applications. In particular, the invention relates to com-
positions containing mineral acid salts of benzophenanthridine
alkaloids mixed with a metal salt, preferably a metal salt
of a mineral acid, although salts of mono-or dicarboxylic
acids, among others, may also be used.
One of the important sources of sanguinarine is a
perennial herb native to North America called Sanguinaria
canadensis Linne (Family: Papaveraceae~ commonly known as
blood root, red root, puccoon, etc. The plant contains
benzophenanthridine alkaloids including sanguinarine,
chelerythrine, and several others. The major alkaloids
present are sanguinarine and chelerythrine. The eighth
edition of the Merck Index lists the alkaloids as sanguinarine,
chelerythrine, protopine and homochelidonine. The pure
chemicals sanguinarine, chelerythrine, and other benzophren-
anthridine alkaloids can be isolated from other plants
besides Sanguinar_a. Also, they are available, even though
very rarely, from some chemical supply houses. Simi-purified
forms of the alkaloids are commercially available, and these
are generally referred to as sanguinarine nitrate and
sanguinarine sulfate. These "salts" are the salts of the
mixed alkaloids of the plant Sanguinaria: mainly sangui-
narine, chelerythrine, and protopine. While few references
can be found in the literature regarding the usage of any
of the pure benzophenanthridine alkaloids, plants containing
such compounds have been used for medical purposes for quite
some time for a wide variety of ailments.
The principle use of sanguinarine up to rece~tly was
a stimulant expectorant to cough syrups containing
" sanguinarine nitrate" .
r~
~7~7~
The use of sanguinarine with thiophosphoric acid
in various animal and human neoplasms is shown in French
patents, number 70-22029 and 2,152,972.
The alkaloid sanguinarine in solution has been shown
to have some antifungal and antiprotozoan properties. The
sanguinarine is applied as an emulsion topically to fungal
infections. The antibacterial activity of sanguinarine has
been found to vary with the attached radicals, and various
salts of sanguinarine have been shown to have some activity.
The hydrochloride and the sulfate salts have been found to
have some activity against certain bacteria at various con-
centrations. Sanguinarine nitrate is reported to have some
weak bacteriostatic action on various types of bacteria.
SUMMAP.Y OF THE INVENTION
The present invention relates to the preparation and
the use of antimicrobial agents, formed particularly from
mineral acid salts of benzophenanthridine alkaloids and a
metal salt and useful in dental preparations, mouthwashes,
rinses, surgical soaps, shampoos, creams, lotionslpowders t
injectables, etc., and other forms of drug preparation and
disinfectants. The mineral acid salts of the benzophenan-
thridine alkaloids may be used in various concentrations with
a metal salt as an antimicrobial agent for use in treating
both human and animal infections and diseases.
Metal salts particularly useful in the formulations
of the present invention include metal salts of halogen acids.
Among these salts are zinc chloride, stannous fluoride, and
sodium fluoride, although any metal salt can be used. This
includes alkali, alkaline earth, and heavy metal fluorides,
chlorides, bromides, and iodides.
Althouth non-toxic metal salts of halogen acids are
preferred for use in the formulations according to the
present invention, it has been found that non-toxic salts
of other acids, such asmineral acids and mono-and dicar~
boxylic acids, are also effective. Examples of acids,
-- 2
74~7
the non-toxic metal salts of which can be used, include
sulfuric acid, nitric acid, and acetic acid.
Glycerol is the preferred vehicle of the formulations
according to the present invention, although other vehicles
that can be used include organic solvents such a propylene
glycol, dimethyl sulfoxide ~DMSO), lower alcohols, and the
like.
It is therefore, among the object and advantages of
the present invention to provide an antimicrobial agent of
mineral acid salts of a benzophenanthridine alkaloid and a
metal salt, useful for topical administration, injectables
and other forms of drug preparations.
Another object of the invention is to provide a
benzophrenanthridine alkaloid salt-metal salt preparation
for treatment of periodontal disease, prevention of dental
caries and similar oral cavity impairmen's.
Still another object of the invention is to provide
drug preparation of a benzophenanthridine alkaloid and a
metal salt, uesful for treatment of ringworm infections,
acne, cold sores and various parasitic infections.
Yet another object of the invention is to provide
a drug preparation of a benzophenanthridine alkaloid and a
metal salt, useful for treatment of scours in animals.
These and other objects and advantages of the
invention may be readily ascertained by the following
description and examples of the preparation of the invention.
The drug preparation of the present invention can be
incorporated in dental preparations, toothpastes, mouth
rinses, surgical and other soaps, vehicles for topical
applications, vehicles for parenteral or intramuscular
injection, and the like.
PREPARATION OF ANTIMICROBIAL AGENT
The pure chemical, either sanguinarine, chelerythrine,
or other benzophenanthridine alkaloids, is dissolved in a
chloroform/methanol mixture and acidified with a mineral
~1774(~7
acid such a hydrocholoric acid. The acidic mixture is
evaporated to dryness and the residue is recrystallized
from ethyl alcohol/chloroform, 50/50.
For use, the mineral acid salt of the benzophenan-
thridine alkaloid is dissolved in either deionized water or
Cl-C6 alcohols, glycerine, propylene glycol, petrolatum, or
other organic solvents at 70 degrees C, and a metal salt or a
salt-forming acid is added to the above solutions. The
preparations generally contain 0.1% by weight and up to 20%
by weight of the benzophenanthridine alkaloid salt, and at
least 1% and up to 60% by weight of a metal salt, with the
remainder being the solvent. The material can be diluted to
the desired concentration, depending on the type of use,
with the solvents listed above~
The benzophenanthridine alkaloid salt is used in
preparations containing 0.01%-10% benzophenanthridine by
weight. The metal salt is present in amounts ranging from
about 2% to about 60%. The lower concentrations are
generally effective in the treatment of most diseases as
explained below.
An example of a basic preparation:
sanguinarine chloride0.3%
glycerine U.S.P. 64.7%
zinc chloride AR. 35.0%
The basic preparation can be varied by using, in place
of sanguinarine chloride, 0.3% of another mineral acid salt
of a benzophenanthridine alkaloid, suchas chelerythrine
chloride.
A second example of a preparation is:
sanguinarine chloride1.0%
glycerine U.S.P. 96.0%
zinc chloride AR. 3.0%
A third example of such a preparati~n is:
sanguinarine chloride 1.0%
glycerine U.S.P. 64~0%
zinc chloride AR. 35.0%
~77~
A fourth example of such a preparation for dental
use is:
sanguinarine chloride1.0%
glycerine U.~.P. 95.6%
zinc chloride AR. 3.0%
stannous fluoride 0.4%
Additional formulations for a basic preparation are
as follows:
sanguinarine chloride1.0%
stannous fluoride 0.4%
glycerine, U.S.P. 98.6%
and
sanguinarine chloride1.0%
sodium fluoride 3.0%
glycerine, U.S.P. 96.0%
Numerous types of diseases were treated in humans and
in animals with the composition of the present invention as
follows:
Example 1 - Canine Ringworm
A 5-10% solution of the first basic preparation was
used, applièd directly to the infected area, one to three
applications as indicated, 48 hours apart.
Example 2 - Feline Ringworm
A 4-8% solution of the first basic preparations was
applied directly to the infected area, 48 hours apart up to
three applications.
Example 3 - Bovine Ringworm
The etiological agent of this involvement is usually
the mold known as Trychophyton album. The duration of the
disease is 4-12 months.
A 30% dilution of the first basic preparation is used,
applied directly to the involved areas, 48 hours apart. Three
applications proved to be adequate in treating the condition
successfully.
Example 4 - Bovine Neo-natal Diarrhea
Twelve animals having diagnosed and confirmed neo-
natal diarrhea were treated with 0.75 grams of the first
basic preparation orally, once per animal, and showed clinical
cure with one exception.
This constitutes an excellent result, considering that
conventional antibiotic therapy currently in use has a much
lower percentage of success.
Sanguinarine chloride and chelerythrine chloride have
strong antimicrobial properties. Zinc chloride has anti-
microbial properties only in high concentrations. It can be
seen from Table I that sanguinarine chloride mixed with zinc
chloride in a l:l ratio, as a rule, did not show a synergistic
effect or even an additive action against most microorganisms
tested n vitro. Further, it was found that in most cases
.the antimicrobial action of the sanguinarine chloride and
zinc chloride mixture depended mostly on the amount of
sanguinarine chloride present in the mixture, and was
relatively independent from the amount of zinc chloride.
TABLE I
Mean inactivating do~s in micro-
grams per milliliter (ug/ml) of
media.
Microorganism ZnCl2 Sanguinarine ZnCl~,& Sang-
Chloride uin ine
Chloride (l:l)
Bacillus subtilis25,000 22 1,000
Escherichia coli6,250 270 500
Klebsiella pneumonia 3,125 540 l,000
Proteus vulgaris12,500 590 l,000
Staphylococcus aureus6,250 70 500
Streptococcus faecalis25,000 393 500
Streptococcus mutans 1,563 161 63
Candida albicans 150
Saccharomyces cerevisiae6,250 20 63
Pseudomonas aeruginosa3,500 7,000 400
~774C~7
A separate test was conducted to determine the
inhibitory concentration of sanguinarine chloride alone.
These concentrations, for microorganisms ln v tro, are as
follows:
100 micrograms per milliliter for Escherichia coli
100 micrograms per milliliter for Candida albicans
50 micrograms per milliliter for Streptococcus mutans
10 micrograms per milliliter for Staphylococcus aureus
It was further found that a concentration of sangui-
narine chloride of 25 micrograms per milliliter caused a 100%
reduction of dental plaque by inactivating plaque forming
microorganisms freshly collected from human dental plaque~
Sanguinarine Chloride compared favourably in vitro to
chlorhexidine (Hibitane ~ , a material used as a standard in
evaluating inhibition of human dental plaque forming micro-
organisms.
However, under in vivo test conditions, sanguinarine
chloride was found to be ineffective against plaque forming
microorganisms.
When sanguinarine chloride was applied to the affected
area of both dogs and humans, repeated treatment with sangui-
narine chloride alone did not reduce dental plaque or
alleviate the symptoms of gingivitis or periodontal disease.
Continuous treatment with sangunarine chloride did not prevent
the accumulation of dental plaque on teeth or prevent the
occurrence of periodontal disease. However, it has been found
that a combination of sanguinarine chloride and zinc chloride
in glycerine is effective in vivo in reducing dental plaque
and the incidence of periodontal disease, and has shown
definite promise in the treatment and prevention of human
periodontal disease.
Results of tests on guinea pigs with induced ringworm
infection, dogs with periodontal disease, and humans with
periodontal disease have shown that glycerine preparations of
sanguinarine chloride plus zinc chloride are far superior for
7 --
~.
~77~
the management of infections ln vivo than either zinc chloride
or sanguinarine chloride alone.
When zinc chloride was replaced with a fluoride salt in
the present preparations, the activity of the preparation
against microorganisms believed to be associated with the
causation of dental caries and periodontal disease ~Stre~-
tococcus mutans), as well as other microorganisms, remained
identical. Data supporting this phenomenon is presented in
the following tables:
TABLE II
Minimum Inhibitory Concentrations
(MIC) in ug/ml
AEROBIC 1.0% sanguinarine 1.0% sanguinar-
MICROORGANISM chloride ine chloride
TESTED 3.0% zinc chlor- 0.4% stannous
ide in glycerol fluoride in
-- glycerol
Escherichia coli 158 160
Klebsiella pneumoniae 158 160
Proteus Vulgaris
Streptococcus mutans79 80
Streptococcus faecalis ~ 20 c20
Staphylococcus aureus ~20 ~20
Pseudomonas aeruginosa 630 1,280
Saccharomyces cerevisiae ~ 20 c20
Candida albicans 79 ~20
. . _
'~.
~77~(17
TABLE III
Minimum Inhibitory Concentrations
(MIC) in ug/ml
1.0% sanguinarine 1.0% sangui-
ANAEROBIC chlorîde narine chloride
MICROORGANISMS 3.0% zinc chlor- 0.4% stannous
TESTEDide in glycerol fluoride in
glycerol
Bacteroides melanino-
genicus 8 8
Eikenella corrodens32 32
. . .
Actinomyces viscosus8 8
Actinobacillus
actinomycetemcomitans 16 16
Capnocytophaga ~ingivalis 8 8
Capnocytophaga sputigena 8 8
TABLE IV
Minimum Bactericidal Concentrations
(MBC) in ug/ml
1.0% sanguinarine 1.0% sanguinarine
chloride chloride
MICROORGANISMS 3.0% zinc chlor- 0.4% stannous
TESTEDide in glycerol fluoride in
glycerol
Escherichia coli 158 160
Klebsiella pneumon1ae 1,261 640
Proteus vulgaris2,522 640
Streptococcus mutans 158 160
Streptococcus faecalis 315 160
Staphylococcus aureus 158 80
Pseudomonas aeruginosa 2,522 2,560
~ ~ =. = . . . . =
Saccharamyces cervisiae 20 20
Candida albicans 79 160
_ g _
X .
11774(~7
The above phenomenon is not what would be expected in
light of the in vitro test results. In most cases sanguin-
arine chloride in vivo was only slightly effective or not at
all. This was quite unexpected considering that in vitro all
the antimicrobial activity of a sanguinarine chloride-metal
salt mixture could be explained by the amount of sanguinarine
present in the mixture.
Controlled clinical tests on 24 male beagle dogs showed
that after four weeks of treatment the dogs treated with the
zinc chloride-sanguinarine chloride-glycerine mixture had the
lowest plaque and gingivitis scores, while the dogs treated
with sanguinarine chloride alone had the highest. The dogs
were treated topically once daily with the respective test
formulation.
The results presented in Table ~ clearly indicate that
the sanguinarine chloride-zinc chloride in glycerine treated
group of dogs had lower gingivitis scores after four weeks of
treatment than did the other groups. Zinc chloride alone was
slightly active, but sanguinarine chloride alone showed no
activity _ vivo at all. This result is unexpected, con-
sidering that, ln vitro, sanguinarine chloride is quite
effective against microorganisms.
-- 10 --
~17~74~7
TABLE V
Oral Clinical Studies - Beagles (dogs)
After
Treatment 0 (start) 4 weeks
Mean Ginyivitis Scores
None 0.475 0.91
0.1% Sanguinarine chloride 0.495 0.942
2.7% Zinc chloride 0.44 0 75
0.1% Sanguinarine chloride with
2.7% zinc chloride in
glycerine 0.418 0.548
Mean Plaque Scores
None 7.478 12.408
0.1% Sanguinarine chloride 8.885 12.89
2.7% Zinc chloride 8.76 10.15
0.1% Sanguinarine chloride
with 2.7% zinc chloride
in glycerine 8.578 7.407
Mean Pocket Depths
None 1.459 1.458
0.1% Sanguinarine chloride 1.415 1.48
2.7% 2inc chloride 1.46 1.37
0.1% Sanguinarine chloride
with 2.7~ zinc chloride
in glycerine 1.445 1.44
Similar results occurred, when the dogs were evaluated
for the return of dental plaques (See Table V for plaque
scores), sanguinarine chloride alone was not active in vivo.
Zinc chloride was slightly preventive but the preparation of
sanguinarine chloride-zinc chloride in glycerine not only
prevented the return and further proliferation of dental
plaque but significantly further reduced it during the four
weeks of treatment.
~7~4Q~
Data also indicate that even the pocket depth.s were
reduced somewhat by the treatment materials containing zinc
chloride in glycerine, or zinc chloride-sanguinarine chloride
in glycerine.
In a clinical test involving twenty volunteer patients
with periodontal disease, it was observed that after treatment
with benzophenanthridine alkaloid chloride-zinc chloride in
glycerine, there was rapid improvement. Inflammation, in-
fection and pockets were eliminated, abcesses ceased, gingival
tone greatly improved, ~issues healed, and in some cases normal
tissue was restored and teeth mobility was reduced.
The clinical studies above showed that sanguinarine
chloride in vlvo was only slightly antimicrobial, very slow
acting, or had no effect on the course of the infection at all.
Zinc chloride in the concentrations required ln vivo to
have antimicrobial action caused blanching of tissue and, in
some cases, chemical burns and other tissue damage. Further,
zinc chloride was found to be slow acting in vivo as an anti-
microbial agent.
Glycerine preparations containing sanguinarine chloride
or other benzophenanthridine alkaloids and zinc chloride or
stannous fluoride or sodium fluoride were fast acting, re-
quiring only one to three applications to clear out infections
rapidly. Further, these preparations did not appear to have
the undesirable side effects of the zinc chloride in the con-
centration necessary to achieve antimicrobial effectiveness.
Example 5 - Human Periodontal Disease
It has been reported that periodontal (gum) disease
affects 2 out of 3 middle-aged ~mericans. Destruction of the
tissue and structures that hold teeth fast in their sockets
accounts for 75% of tooth loss after the age of 40. Most
cases of periodontal disease are the result of neglect and
can largely be prevented by a regular program of thorough
hygiene. In the most common type of periodontal disease,
the three chief culprits are bacteria, calculus (tartar), and
food debris.
- 12 -
X
~'774(~7
The invention has been used by dentists in clinical
management of over forty cases of various types of human
periodontal disease.
In some cases, even a single treatment brought major
improvements in the conditions of the diseased gums. Clinical
cure resulting from the treatment was quite apparent and
included: elimination of inflammation, normal tissue tone
restored, pockets were eliminated, infections were cleared up,
mobility was reduced, and gingival tone was greatly improved.
The materials and methods used were the following:
1. Undiluted Paste for Packing:
In cases of widespread tissue involvement, undiluted
preparation was used in quantity sufficient (q.s.) to "cover"
or "pack" the infected or inflamed areas of gingival tissues~
The clinical procedure consisted of two treatments
approximately two weeks apart, with the application of not
less than l.0 mm thickness of the basic preparation to the
diseased periodontium. In cases where undiluted material was
used, it was either applied topically with a spatula or 0.25
ml was pressed through a 22 guage needle attached to a 1.0 ml
pressure syringe. The material was introduced into the ging-
ivae to the attachment (q.s.) to fill the pocket and the
medication was left in place for 10-15 minutes.
2. String Technique:
Cotton String Saturated with the Drug Preparation.
In cases where individual teeth were to be treated, an
ordinary soft cotton string, sterilized before use, or
"gingipak" was used (gingipak contains racemic epinephrine
hydrochloride 8-100 solution and 1% benzyl alcohol as
preservative).
Cotton strings 1-1.5 cm in length were saturated with
undiluted material by using a spatula and precut pieces of
string which were then impregnated on a dentist's mixing pad.
- 13 -
~7~07
These strings, when properly impregnated with the preparation,
weigh approximately 35 mg/cm.
One to three strings were used per tooth depending upon
the circumference of the tooth. Up to three strings were some-
times used for a total of approximately 10.5 mg of the pre-
paration. Inpregnated strings were left in place 10-15 minutes.
Dental floss or similar substrate, such as synthetic
hollow fiber string, can be impregnated with the basic pre-
paration for use in treating the teeth and gums.
3. Dilution of Paste for Irrigation
In addition to "packing" of the periodontium or employ-
ing the "string technique" on individual teeth, irrigation was
often used concurrently as part of regimen. Generally, the
diseased periodontium was first packed with undiluted pre-
paration or the teeth were individually treated by the string
technique described above. These methods of treatment were
followed by irrigation with a suspension of the preparation
in either water or glycerine. Suspensions were prepared to
contain 1 part of full strength material to 1 part of gly-
cerine, or 1 part of water depending on whether a glycerine or
water suspension was desired. The final suspension was V/V
mixture of 1 part material to 1 part diluent. Irrigation was
accomplished by filling a 7.0 ml syringe to contain 420 mg of
the suspension. For treatment of individual teeth, a total of
1.0 ml of material in suspension was used to irrigate the
buccal, lingual, and interproximal areas about the tooth.
Where indicated, all teeth were irrigated with 840 mg of
the suspension contained in two syringes.
Example 6 - Dental Caries
Preparations containing about 0.3~ sanguinarine
chloride and about 35% zinc chloride were used on seven
patients with dental caries. Decay was removed from the
teeth with a spoon excavator leaving a layer of carious
- 14 -
~3L7~9~0~
tissue about 1 mm to 1 1/2 mm in depth. The antimicrobial
preparation was placed over the remaining decay, about 56.1 mg
of preparation, with a ~ollenbeck carver and uniformly applied
over the decayed area with a piece of cotton held in cotton
tweezers. Intermediate restorative material (IRM) was used as
a temporary restoration to seal the material in the cavity
preparation.
After several weeks (6 weeks) specimens for bacteri-
logical and histological studies, including electron micro
scopy, were undertaken.
The conclusions of the investigators were that the
preparation could be considered a cariostatic agent. Further,
the material may enhance sclerotic dentine formation, thus
forming a hard protective floor between the carious lesion and
the pulp.
The benzophenanthrine chloride-zinc chloride-glycerine
composition can be used in conjunction with a fluoride-
providing compound. These compounds are characterized by
the ability to release fluoride ions in water and by sub-
stantial freedom from reaction with other compounds present
in the oral preparation. Among these materials are inorganic
fluoride salts such as suitable alkali metal, alkaline earth
metal, and heavy metal salts. Alkali metal and tin fluorides,
such as sodium and stannous fluorides, and mixtures thereof,
are preferred.
The amount of the fluoride-providing compound is
dependent to some extent upon the type of compounds, its
solubility, and the type of oral preparation, but it must be
a nontoxic amount. Any suitable minimum amount of such com-
pound may be used, but it is preferable to employ sufficient
compound to release from 0.005~ to 1~, most preferably about
0.1% by weight of fluoride ion. Typically, in the cases of
alkali metal fluoride and stannous fluoride, this component
1~774~7
is present in an amount up to 3% by weight, based on the
weight of the preparation, and preferably in the range of
from 0.05% to 1%.
Example 7 - Treatment of Animal Skin Tumors
Equine sarcoid (a locally malignant connective tissue
tumor found on the skin of horses similar to fibrosarcomas
which occur in other animals and man) are probably of viral
origin transmissible between horses. These tumors are in-
vasive and commonly result in sufficient debilitation of the
horse that euthanasia is performed.
Surgical removal is the accepted means of therapy.
Recurrence of the tumor is the rule rather than the exception.
Sixty tumors were treated with the preparation employing
the following treatment method:
~ Either spread the first basic preparation on the tumor
with a wooden applicator, bandage and observe in 24 hour
intervalsf~or place about 3 mm thickness of the preparation
on a Telf~3pad covering an area slightly larger than the base
area of the tumor, securing the Telfa pad on the tumor
with a bandage. Observe in 24 hour intervals. Repeat appli-
cation if needed after 48 hours.
Three to four or more applications may be necessary in
cases of long duration and if the tumor is deep. Recurrence
of the tumor occurred in only 25% of the cases.
In all cases, the tumors were removed by the excising
action of the preparation and in no case thus treated did a
post treatment infection (secondary infection) develop. This
was true despite the fact that no covering was maintained on
the lesions after the tumor came out, nor was any form of
other antibacterial medication applied.
After the second or third application, the tumor
sloughed out, a very firm scab formed on the tumor base, and
healing proceeded under scab. The healing of the would was
slow, but uneventful. Scars remaining were quite small.
- 16 -
~L1774~7
Example 8 - Equine Dermal Fibrosarcomas
Seventy-five cases of equine dermal fibrosarcomas were
treated topically as described in Example 7. Only 20~ of the
tumors recurred within the one year observation period. While
the cure rate is somewhat less than 80~, this compares
favorably with the approximate 55% cure rate of this tumor
following surgical removal and radiotherapy.
Example 9 - Bovine Squamous Cell Carcinoma
Seven Hereford cattle diagnosed with the disease were
treated with about 15 grams of the first basic preparation
pasted right over the lesions. Improvement of clinical con-
dition followed in all cases.
Example lO - Treatment of Squamous Cell and Basal Cell
Carcinoma in Animals of Non-bovine Species
Canine - Histologically confirmed squamous cell car-
cinoma of both eyes, five applications with the first basic
preparation in five days; in two weeks time, tumors reduced
in size and were sloughing.
Equine - Histologically c~nfirmed basal cell carcinoma
was treated by six direct topical applications of the pre-
paration locally, once daily. In three weeks, most visible
tumors were gone.
Equine - Histologically confirmed large hair follicle
carcinoma on chest; tumor removed by surgery; ten topical
applications of the preparation into the open wound. In three
months lesion healed and no recurrence of the tumor was noted.
Example ll - Treatment of Human Skin Tumors
~ ascaI Cell Epithelioma:
A team of qualified dermatologists and surgeons treated
sixty patients with diagnosed histologically confirmed basal
~'7~407
cell epithelioma (basal cell carcinoma).
The first basic preparation was directly applied
topically to the involved area and held in place with a
bandage. The lesions were observed in 48 hour intervals and
the preparation was reapplied if it was indicated. In most
cases, 2-3 applications were adequate. Clinical cure followed
the treatment in all cases and no reappearance of the tumors
was noted over a four year period.
Example 12 - Squamous Cell Carcinoma
Sixteen cases of histologically confirmed squamous cell
carcinoma were treated by the same method used in Example 11.
In all treated cases, clinical cure followed treatment and
only in one case was tumor recurrence noted within four years.
Example 13 - Antimicrobial Activity
- In order to illustrate the an~imicrobial effectiveness
of the composition according to the present invention, a series
of _ vitro and in vivo tests were performed. These tests were
designed to demonstrate the lowest concentration of sanguin-
arine chloride that will inactivate microorganisms.
Findings were as follows:
On the average a 22 ug/ml sanguinarine chloride solution
will inactivate cultures of the bacteria Bacillus subtilis, a
270 ug/ml solution cultures of Escherichia coli, a 540 ug/ml
solution cultures of Klebsiella pneumoniae, a 590 ug/ml
solution cultures of Proteus vulgaris, a 70 ug/ml solution
cultures of Staphylococcus aureus, a 393 ug/ml solution
cultures of Streptococcus faecalis, and a 161 ug/ml solution
cultures of Streptococcus mutans (see Table I).
Further on the average a 150 ug/ml and a 20 ug/ml
solution also inactivates the cultures of yeasts Candida
albicans and Saccharomyce_ cerevisiae. A considerably lower
- 18 -
~L~77~(~7
concentration than the inactivation concentration of the drug
preparation is useful as a growth inhibitor of the same
organism.
In addition, on the average a concentration of san-
guinarine chloride presented in Table VI was sufficient per
cubic centimeter of media, to inhibit growth of some fungi
known to belong to the group of ringworm producing organisms.
TABLE VI
Mean Inhibiting Dose of Sangui-
Microorganisms narine Chloride in ug/ml of Media
Microsporum canis 867
Microsporum nanum 650
Trichophyton mentagrophytes 900
Trichophyton schoenleini 467
Trichophyton terrestre 467
. _ _
Trichophyton vanbreuseghemi 750
The basic preparations were, on numerous occasions,
also tested on a variety of animals and also humans infected
with ringworm and athletes foot fungus. It was found that
preparations according to the present invention will consid-
erably hasten recovery from ringworm infection and facilitate
rapid healing of lesions.
Results of clinical tests run by numerous dentists
and dental institutions confirm that regular application of
the composition of the instant invention to teeth surfaces
will reduce the incidence of dental caries and the use on
gums either by packing or irrigations will prevent or rapidly
cure periodontal disease or microbial infections of the gums
and surrounding tissues.
Application of the compositions of the present in-
vention to cold sores immensely hastens the healing process.
Cold sores dry up and heal in a few days.
-- 19 --
~74~7
When animals suffering from scours are treated in-
ternally with preparations according to the present invention,
the recovery rate exceeds that of the antibiotics commonly
used for the purposes of treating scours.
- 20 -
.,,,~ ~
