Note: Descriptions are shown in the official language in which they were submitted.
104Z805
This invention relates to compositions for oral ~ -
hygiene.
It is currently believed that one factor in the form-
ation of dental caries is the presence of food residues
trapped in the interstices between the teeth, and in -
pits and fissures inthe teeth themselves, it being
theorized that carbohydrates in such food residues are
bacterially decomposed to lactic acid whlch promotes de-
calcification of dental material. The acid-producing
bacteria are known to adhere firmly to the surface of
the teeth as a thin film, giving rise to dental plaque.
The formation and adhesion to the teeth of dental plaque
is also thought to be a precursor of tartar formation and
of inflammations of the gum and periodontal tissue, such
as pyorrhea alveolaris and periodontitis. Preventing
the formation of dental plaque, or removing it, is there- ;
fore regarded as an important factor in oral hygiene.
It has now been found that a strain of Actinomyces
viscosus isolated from human dental plaque proudces levan, -
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an exocellu~rfructan, int he presence of sucrose, and that
the levan is implicated inthe formation and adhesion of
the plaque on the surface of the teeth. Furthermore, it
has been discovered that fructan-degrading enzymes can
be used to help diminish the formation of dental plaque,
or to assist in removing it from the teeth, and thus ~
improve oral hygiene. For this purpose, the invention pro-
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vides compositions for dental and oral use, comprising a
fructan-degrading enzyme and an orally acceptable excipient.
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According to one aspect, the compositions of the invention
may take any of the forms normally intended for cleaning, antiseptic
or emollient purposes in the mouth: for example, tooth-powders,
tooth-pastes, mouth-washes, gargles, dental lotions and ointments,
chewing-gums, tablets, and mouth or throat pastilles or troches.
These compositions of the invention comprise a fructan-degrading
enzyme and an edible solid excipient, a dental detergent, an anti-
septic, an emollient, a demulcent, or a chewing-gum base.
In accordance with another aspect of the invention, the
fructan-degrading enzyme can be incorporated into an edible compo-
sition, comprising a foodstuff or beverage. For example, the
fructan-degrading enzyme can be incorporated into soft drinks or
confectionery products, tn combat the formation of dental plaque
otherwise resulting on eating these products.
The fructan-degrading enzyme used in the compositions of
the invention is one produced by a micro-organism of the genus
Talaromyces, ~upenicillium, Sporotrichum, Sordaria, Chaetomium,
Gelasinospora, Stachybotris or Actinomyces, in accordance with the
process of Canadian Patent No. 1,003,351. Although all fructan-
degrading enzymes can be used in the compositions of the invention,it is preferable to use
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those of the liquefying type (i.e. endo-enzymes), which can chop up
the levan molecule anywhere along its length, rather than those of
the saccharifying type (i.e. exo-enzymes), which degrade it step-
wise from the ends. Enzymes of the so-called "levanase type",
which cut ~-(2 -~6) linkages in the main chain of the levan molecule
and enzymes of the so-called "inulase type", which cut ~-(2 >1)
linkages, are both effective in their own right; but it is even
better to use a combination of two or more different types of
highly active fructan-degrading enzyme.
In addition to containing one or more fructan-degrading
enzymes, the compositions of the invention may optionally also con-
tain one or more other enzymes, such as a dextranase (3.2.1.11),
a-amylase (3.2.1.1), protease (3.4.4.16 and 3.4.4.17) or lipase J
(3.1.1.3) in order to supplement their dental cleaning activity.
The concentration of fructan-degrading enzyme used in the
compositions of the invention can vary widely, depending upon the ~-
type of composition, and hence on its expected residence time in
the mouth and frequency of use. A unit dosage of the composition
can generally contain from 1 to 3,000, preferably from 1 to 300 and
~0 most preferably from 10 to 100, activity units of fructan-degrading
enzyme. The compositions should preferably be used several times
daily, suitably with a total daily dosage corresponding to about
1,500 activity units of fructan-degrading enzyme. Naturally, it is
preferable for the compositions to be retained in the mouth for as
long as possible.
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The activity of the fructan-degrading enzyme is
determined by adding it to l ml of M/20 citric acid buffer
(pH 5.4) containing 0.5% by weight of fructan (from Aero-
bacter le _nicum). The mixture is kept at 40 C for 30
minutes, to liberate reducing sugar, which is determined
as fructose with 3,5-dinitrosalicylic acid. The"activity
unit" of the enzyme is defined as that quantity which
liberates 1 mg of fructose in 60 minutes.
The ingredients uscd in the compositions of the
invention, apart from the enzyme or enzymes, will depend
upon the precise form of composition. Any of the ingre-
dients normally used for the types of formulation already
listed may also be used in the compositions of the inven-
tion; and, apart from ingredients such as the detergent,
antiseptic, emollient or demulscent, or the chewing-gum
base, the compositions may also contain conventionally
used additives, such as flovouring agents, perfumes and
sweetners. In general, the fructan-degrading enzyme may
be formulated with any orally acceptable excipient, depending
upon the intended form and use of the composition.
The beneficial effect of fructan-degrading enzyme
in reducing the formation of dental plaque by oral micro-
organisms can be demonstrated in the laboratory by the
methods of Jordan et al. Archs. Oral Biol., 11 739 (1966)
and of Onisi et al. Journal of Dental Health, Japan, 20
292 (1971). In order to simulate experimentally the in
vivo formation of dental plaque, pure cultures were pre-
pared using the
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1042805
following media and micro-organisms~
Med;a: (1) the basic medium of Jordan et al. J.
Dent. Res., 39, 116 (1960)
(2) the basic medium (1) containing 10
activity units per 10 ml of fructan-
degrading enzyme (from Talaromyces
flavus var. flavus)
Micro-organism: (1) Actinomyces viscosus, Onisi No. 39 ;~
strain, isolated from human dental
plaque (similar to Actinomyces
viscosus from rats and hamsters).
(2) Actinomyces viscosus, Onisi No.42
strain, (isolated as above).
Test-tubes containing 10 ml portions of the above mèdia,
which were freshly prepared daily, were sub-cultured with
0.2 ml of cultures of the above micro-orgaisms, and each
tube was fitted with a rubber stopper carrying a number 20
gauge stainless-steel wire which dipped into the medium.
This procédure was repeated for 10 days, and the dental
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plaque sticking to the stainless-steel wire was observed -`
with the naked eye. It was found that the f~`rmation of
dental plaque in the portions of medium (2) containing
fructan-degrading enzyme was greatly inhibited by com-
parison~,with the basic medium (1). Moreover, when the
wire carrying the dental plaque formed in the basic med-
ium (1) was dipped into a solution containing 10 activity
units/nl of fructan-degrading enzyme, the plaque came off -
the wire in a few hours; while no change was observed when
the wire carrying dental plaque was dipped into plain
water, or into a solution of the enzyme inactivated by
heating. These experiments demomstrate the effectiveness ;
of the fructan-degrading enzyme for preventing the
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104Z8~5
formation of dental plaque and for removing it.
The invention is illustrated by the followng Examples,
in which percentages are given by weight and the "activity
units" are as hereinbefore defined.
Example 1 Tooth-paste
Dibasic calcium phosphate 50%
Glycerol 20%
Sodium lauryl sulphate 2.5%
Spearmint oil 2.5%
Gum tragacanth 1.0%
Saccharin 0.1%
23 9%
Water
A fructan-degrading enzyme, derived from Talaromyces
flavus var. flavus, was incorporated into a tooth-paste
having the above composition, at the rate of 10 activity
units per gram of the paste, and the mixture was uniformly
blended to form an enzyme-containing tooth-paste. ~ -
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Example 2
Mouth-wash
Sodium carboxymethylcellulose 4.0%
Sodium lauryl sulphate 2.0%
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Glycerol 104Z8~5 30%
Perfume 0.5%
Water 63.5%
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A fructan-degrading enzyme, derived from Eupenicillium
javanic _ , was incorporated into a mouth-wash having the
above composition, at the rate of 10 activity units per
gram of the wash, and was uniformly dissolved therein to
obtain an enzyme-containing mouth-wash.
Example 3 :::
Chewing-gum ~ :
Polyvinyl acetate 20% -
Butyl phthalylbutylglycolate 3%
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Polyisobutylene 3%
Microcrystalline wax 2%
Calcium carbonate 2%
Glucose 69%
Perfume 1%
A mixture of fructan-degrading enzymes, derived from
Chaetomium subspirale and Eupenicillium javanicum, was
incorporated into a chewing-gum base having the above comp-
osition, at the rate of 10 activity units per gram of the
gum, and the mixture was uniformly blended to form an :
enzyme-containing chewing gum. I
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Example 4 1~4~S
Chewable tablets
Mannitol 65~
Soluble starch 30%
"Avicel" microcrystalline cellulose (a
trademark of Asahi Chemical Industry Co.
Ltd., Japan) 2.5%
Food pigment trace
"Aerosil 200" colloidal silicon dioxide
(a trademark of Degussa, Germany) 0.6%
Magnesium stearate 0.6%
Saccharin 0.8%
"Orange-microne" perfume (a trademark
of Takasago Perfumery Co. Ltd., Japan)0.3%
A mixture of fructan-degrading enzymes, derived from
Sordaria humana and Talaromyces flavus var. flavus, was incorporated
into the above chewable tablet base, at the rate of l00 activity
units per gram. The mixture was uniformly blended and tabulated,
to produce enzyme-containing chewable tablets.
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