Solid composition for the oral administration of dyes and diagnostic use thereof
PRIOR ART
The endoscopy is an exceptionally important diagnostic technique for the diagnosis of
inflammatory, ulcerative and neoplastic pathologies of the gastrointestinal tract.
Actually, the endoscopy allows observing ~ from inside the lumen - the state of
preservation and development of the mucosa that covers the gastrointestinal cavity, as
well as the surface spraying thereof, the presence of deformations and/or ulcerations.
More and more powerful and sophisticated endoscope probes have enabled to improve
this technique considerably; the progress of the used materials also allowed improving
performance in terms of illumination and resolution power.
More recenll y there has been an improvement of the conventional diagnostictherapeutic
aspects due to the use of image magnification and vital dyes, useful for
locally developing a contrasting colour capable of amplifying the resolution diagnostic
power of the conventional technique. The use of dyes in endoscopy lead to coining the
term "chromoendoscopy" for describing this diagnostic procedure, particularly useful
for identifying suspicious areas for degenerative characteristics.
The use of colouring is generally adopted after completing the endoscopic analysis
during the step of withdrawing the endoscopic probe, and after accurately cleaning the
mucosa tract to be examined; currently, the dye is applied to the mucosa by spraying a
small volume of a solution averagely concentrated with dye, using a catheter or
capillary pipe directly inserted into the cavity of the endoscopic probe.
The diffusion of the dye and absorption thereof by the vital cells markedly differentiates
the cells with normal vitality from those in the advanced replication stage, for example
characteristic of neoplastic cells.
The dyes usually used are mainly, but not exclusively, the following: methylene blue,
congo red, carmine indigo, and/or toluidine blue.
Methylene blue and toluidine blue are uniformly absorbed by the whole intestinal
mucosa, while, in case of an inflammatory process, absorption thereof by the mucosal
cells tends to reduce as the phlogosis worsens. Due to this characteristic, the two dyes
are also useful in the step of remission of the inflammatory processes and in the
differential diagnosis between pseudopolyps and true polyps. Also carmine indigo has a
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similar action, finding application in the long duration inflammatory forms and with the
aim of highlighting carpet lesions, which can contain tumoral forms, which are difficult
to detect with the conventional endoscopy in absence of contrasting colour.
Within the procedure for applying the dye, it should be observed that use thereof reveals
several practical problems that are difficult to resolve due to the considerable
application difficulty. First and foremost the pharmacy of the institute where the
endoscopy is performed should be capable of preparing solutions with concentrations
generally comprised between 0.1% and 1% of the dye; then, the endoscopic probe
should be provided with a channel for inserting the capillary catheter which carries the
solution up to the point of application; then the dye should be dispensed uniformly so as
to cover the mucosal surface subject of the evaluation. The need for the simultaneous
presence of these precise conditions contributes to the difficulty of executing the
chromoendoscopy procedure, which is exclusively carried out by the best diagnostic
centres up to date, with extensive defections by hospitals and nursing homes specialized
in gastroenterology.
Furthermore, it should be taken into account that the use of a solution to be locally
sprayed on the mucosal wall does not entirely solve the problem regarding forms still
latent, in that too small to be detected, as well as the degenerative processes of the
digestive system.
Thus, there arises the need of providing a simple and safe use of a dye in diagnostic
endoscopies, through a suitable means of administration also capable of guaranteeing a
homogeneous and complete distribution for an ideal effect of the dye in the area in
question.
Now, it has been surprisingly discovered that a solid composition for oral administration
allows formulating one, or more, dyes which can thus reach the desired site providing
the contrasting image required for endoscopic diagnostic evaluation.
DESCRIPTION
Thus, the present invention is aimed at providing a solid composition for oral
administration containing at least one dye suitable for the preparation and evaluation of
the endoscopic diagnostic analysis, and at least one physiologically acceptable
excipient.
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Physiologically acceptable excipients according to the invention are prefcrabl y
cxcipients useful for reaching a mass identifiable uniquely, in terms of quality and
quantity, and which can be easily administered through the oral cavity.
In particular, the composition of the invention is intended for oral uptake, before being
subjected to the endoscopic diagnostic analysis, during or at the end of the procedure for
preparing such analysis.
Thus, an object of the present invention is represented by a solid composition, for oral
administration, containing at least one dye in association with at least one
physiologically acceptable excipient, intended for allowing early local identification of
pathologic forms at the gastrointestinal mucosa level, preferably precancerous fonns or
mucosal areas with high inflammation rate.
The composition of the present invention can be instant-release or controlled-release
type, preferably of the controlled release type capable of selectively carrying the dye in
the regions subject of the analysis, thus preventing dispersion thereof into areas not
subject of the analysis.
The expression "instant-release" is used to indicate a composition capable of
disintegrating quickly and dissolving in the gastric cavity, simultaneously releasing the
entire dye contained therein.
The instant-release composition of the invention preferably comprises at least one dye
in association with physiologically acceptable excipients, technologically indispensable
to guarantee the quick disintegration and dissolution of the form in the gastric cavity;
more preferably, are used the so-called super disintegrants, i.e. polymeric substances
capable of swelling on contact with aqueous fluids and triggering a hydrodynamic
tension within the pharmaceutical form which leads to the breakage thereof into
fragments with the ensuing considerable increase of the surface/volume ratio and, thus,
to a more rapid dissolution of the dye/s contained in the administration form.
Suitable super disintegrants are preferably selected from among modified starches,
modified celluloses, polymers or cross-linked copolymers (such as, for example, crosslinked
polyvinylpyrrolidone) or mixtures thereof.
The instant-release composition of the invention may also comprise an outer coating,
preferably selected from among polymers and copolymers of the acrylic or mcthacrylic
acid, alkyl or hydroxyalkyl cellulose derivatives or mixtures thereof.
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The possible presence of such an outer coating is useful for avoiding the coloration of
the mucosa of the mouth and/or of the throat during the uptake and swallowing by the
patient.
The expression "controlled release" of the invention is used to indicate a composition
capable of releasing the dye in a selective site-time manner, i.e. progressive in the areas
of interest. Thus, such expression comprises the "rapid, delayed or modified" release
definition.
The technology suitable for the formulation of controlled release composition of the
invention can be selected from among the matrix technologies and the reservoir
diffusion technologies known in the sector.
Preferably the controlled release solid oral composition of the invention is formulated
according to the multimalrix technology commercially known under the trade name
MMX@, described in the international patent applications W000/76481 and
W000/76478, incorporated herein by reference.
According to a preferred embodiment of the invention, the controlled release solid oral
composition comprises at least one dye and a multimatrix structure containing:
a) a matrix which consists of lipophilic compounds with melting point below 90°C,
and optionally amphiphilic compounds, in which at least one dye is at least partly
incorporated;
b) an outer hydrophilic matrix in which the lipophilic matrix, and optionally the
amphiphilic matrix are dispersed;
c) optionally other physiologically acceptable excipients;
d) optional gastro-resistant coating.
According to a further embodiment, the composition containing at least one dye
compnses:
a) a matrix which consists of lipophilic compounds with melting point below 90°C and
amphiphilic compounds in which said at least one dye is at least partly incorporated;
b) an outer hydrophilic matrix in which the lipophilic/amphiphilic matrix IS
dispersed;
c) optionally other physiologically acceptable excipienls;
d) optional gastro-resistant coaling.
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According to another embodiment, the composition containing at least one dye
compnses:
a) a matrix which consists of lipophilic compounds with melting point below 90°C,
in which said at least one dye is at least partly incorporated;
b) an outer matrix which consists of hydrophilic compounds and optionally
amphiphilic compounds, in which the lipophilic matrix, is dispersed;
c) optionally other physiologically acceptable excipients;
d) optionally a gastro-resistant coating.
Suitable lipophilic compounds in the present invention are preferably selected from
among saturated, unsaturated or hydrogenated long chain alcohols, saturated or
unsaturated or hydrogenated fatty acids, salts thereof, esters or amides, mono-, di- or
triglycerides of fatty acids, polyethoxylated derivatives thereof, waxes, ceramides,
cholesterol, cholesterol derivatives or mixtures thereof.
Suitable amphiphilic compounds in the present invention are then preferably selected
from among polar lipids of type or 11 (lecithin, phosphatidylcholine,
phosphatidylethanolamine or a mixture thereof), ceramides, glycol alkyl ethers (such as
for example, diethylene glycol monomethyl ether), alkyl sulfate or sulfosuccinate salts
or mixtures thereof.
Suitable hydrophilic compounds in the present invention are preferably compounds
forming hydrogel (i.e. compounds which form hydrogel on contact with aqueous
solvents), more preferably selected from among polymers or copolymers of the acrylic
or methacrylic acid, alkyl vinylpolymers, alkyl celluloses, hydroxyalkyl celluloses,
carboxyalkyl cellulose, modified or plurisubstituted celluloses, polysaccharides,
dextrins, pectins, starches, complex starches and starch derivatives, alginic acid,
synthetic rubber, natural rubber, polyalcohols or mixtures thereof.
Suitable gastro-resistant coating according to the invention is preferably selected from
among polymers of the acrylic or methacrylic acid, copolymers of the acrylic or
methacrylic acid, cellulose derivatives (such as for example cellulose acetate phthalate)
hydroxybutyrate-based polymers, shellac or mixtures thereof. Such gastro-resistant
coatings of the invention can also be combined with plasticisers, opacifiers, dyes or
mixtures thereof.
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While the oral administration of instant-release solid pharmaceutical forms allows
obtaining a coloration of the first portion of the digestive tract, such as oesophagus or
stomach, the administration of controlled release compositions of the invention actually
allows releasing the dye contained in the composition precisely starting from the
gastrointestinal segment intended to be subjected to endoscopic evaluation.
Preferably the composition of the present invention is formulated in form of tablet,
capsules, granules, microgranules or pellets. The capsule form according to the
invention may in turn contain granules, microgranules or pellets.
More preferably, the composition of the invention may be formulated in form of gaslroresistant
tablet or in form of a capsule containing granules, microgranules or gastroresistant
pellets.
Furthermore, the composition of the invention may be formulated in a double layer
fom1, preferably a double layer tablet.
More precisely, instant-release compositions to be administered a few minutes before
carrying out the endoscopy analysis using a glass of aqueous liquid are preferable for
gastroscopy.
The aqueous liquid is used for facilitating the dissolution of the composition, thus
forming- in loco- a coloured solution that allows the dye to homogeneously reach the
mucosa which covers the digestive cavity, and to be absorbed or not by the cells of the
mucosa covering the stomach.
In case of gastroscopic analysis, the composition of the invention is thus preferably in
form of instant-release tablet or capsule.
The same target may be attained in the small or large intestine, due to the use of forms
of controlled or targeted release oral administration of the dye; in particular, a
composition coated with a thin film of contro11ed release gastro-resistant polymers is
preferred for an environmental pH of about 5 for the small intestine.
In case of endoscopic analysis on the small intestine, the composition of the invention is
thus preferably in form of controlled release capsule or tablet, with the presence of a
gastro-resistant coating more preferably selected from among mixtures of acrylic and
methacrylic copolymers of type A (Eudragit L, or RL, for example).
Even in case of colon endoscopy, it is preferable to use forms of oral administration in
solid oral form, coated with gastro-resistant substances, preferably tablet or capsule.
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Such gastro-resistant substances are preferably selected from among acrylic and
methacrylic copolymers of type A, type B (such as for example those commercially
known under the trade name Eudragit S or RS), and/or mixtures based on cellulose
acetate phthalate, insoluble in an acid environment which become soluble when the pH
is neutralized and acquires a value of about 7. A similar event also occurs when the
intestinal transit leads the cud to pass through terminal ileum or through the ileocecal
valve. Obviously, in the latter case, considering the fact that the cud takes at least 3-5
hours to complete the transit in the small intestine and an undetermined period of time,
ranging from a few minutes to a few hours, for the gastric emptying, the administration
of the dye composition should be carried out suitably in advance with respect to the
endoscopic analysis, generally for a period comprised between 4-24 hours, so as to
allow the dissolution of the dye in situ, the formation of a concentrated solution within
the colon lumen due to the intestinal fluid present therein, and the diffusion of the dye
on the mucosa for a period of time in which the endoscopic probe is introduced into the
intestinal cavity.
In order to allow a homogeneous coloration of the luminal membrane of all colon
regions, from the ileocecal area to the ascending, transverse, descending, sigmoid and
rectal colon, the release of the dye should not be instantaneous but progressive and in
line with the advancement of the composition.
Given that the transit time of the colon tract is once again very variable, but estimated at
least 8-16 hours, it is clear that a controlled release composition with the dye being
released in vitro in about 6 -8 hours constitutes the best system to allow a homogeneous
coloration of the entire membrane to be analysed endoscopically and, thus, to obtain the
best result in terms of diagnostic evaluation.
Useful dyes according to the present invention are preferably selected from among
congo red, carmine indigo, methylene blue, toluidine blue or mixtures thereof.
However, according to the invention also other biocompatible dye substances can be
used, as long as they are provided with a toxicity profile that does not represent an
obstacle to oral systemic administration thereof.
Therefore, the amount of dye that can be used for maximising the structural contrast of
the mucosal cells depends:
- on the inherent capacity of the dye to induce the coloration of the vital cells,
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-on the period of time that this coloration should be kept in contact with the cells and
- on the massive presence of the liquid for washing the mucosa swallowed during the
step of preparing the colonoscopy.
Actually, such parameters may vary the amount of dye from a few milligrams to a few
grams of substance, divided into one or more solid oral compositions to be swallowed
before or during the step of preparing the endoscopic procedure, or at the end of the
procedure.
Preferably, the solid oral composition of the invention comprises at least one dye in an
amount comprised between 10 mg and 1500 mg (1.5 g), per single composition, more
preferably between 50 mg and 1200 mg (1.2 g) per single composition.
Said at least one dye according to the invention may also be comprised in an amount
between 2 (0.002 g) and 1000 mg (1 g), more preferably between 10 mg (0.()1 g) and
500 mg (0.5 g) per single composition.
Said at least one dye according to the invention may also be comprised in an amount
between 20 (0.02 g) and 500 mg (0.5 g), even more preferably between 25 (0.025 g) and
400 mg (0.4 g), per single composition.
According to a preferred embodiment said at least one dye is contained in the solid
composition of the invention in an amount equivalent to about 25 mg (0.025 g).
According to a further embodiment said at least one dye is contained in the solid
composition of the invention in an amount equivalent to about 50 mg (0.05 g).
According to another embodiment said at least one dye is contained in the solid
composition of the invention in an amount equivalent to about 200 mg (0.2 g).
According to a preferred embodiment of the present invention in case of gastroscopy the
administration is provided for about 30 minutes before the execution of one or more
compositions of the invention, preferably instant-release tablet or capsule.
According to a further embodiment of the present invention, in case of endoscopy of the
small intestine there is provided for the administration of one or more compositions of
the invention, preferably a controlled release tablet protected by a gastro-resistant
coating to prevent early dispersion of the dye in the gastric area not intended to be
subjected to the endoscopic evaluation.
According to a further preferred embodiment of the present invention, in case of
colonoscopy there is provided for the administration of one or more compositions of the
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invention, preferably a controlled release tablet so as to prevent the dye from being
dispersed into areas of the digestive tract not intended to be subjected to colonoscopy,
such as for example the stomach, duodena and jejunum.
For the preparation of controlled release compositions, one or more dyes are preferably
formulated alongside substances capable of imparting progressive or massive or
controlled or prolonged dissolution properties to the fonnulation; in addition, the
fonnulalion is coated with substances capable of dissolving solely upon reaching a
specific pH, generally comprised between 5 and 7, typical of the section subject of the
intestinal endoscopic evaluation.
Upon reaching the intestinal section of interest, characterised by a specific pH value at
which the gastro-protective coating starts dissolving, it is important that the dissolution
of the dye be controlled in terms of speed so as to ensure that it occurs within the time
indispensable to the intestinal transit, generally comprised between 4 and 24 hours.
Various formulation technologies can be used according to the invention for such
purpose.
As mentioned previously, the main and known technologies for obtaining a colon
release, such as the use of reservoir systems or diffusion or hydrophilic matrix
structures, can be applied for preparing the controlled release composition of the
invention; the multi-matrix technology which exploits a sequence of hydrophilic,
lipophilic and amphiphilic matrices for obtaining a result as described above is used in a
preferable application of the invention. In a typical application of this multimatrix
technology, the dye/s is/are first mixed or granulated with the material capable of
forming a lipophilic matrix, in the presence of one or more amphiphilic substances with
surfactant properties, and lastly this matrix of powders, at any degree of aggregation, is
inserted into a dominant structure formed by polymers or copolymers of the hydrophilic
type, also known as hydrogels, in the anhydrous state or with low residue moisture
value.
Alternative! y, still according to a typical application of this technology, the dye/s should
be first mixed or granulated with the material capable of forming a lipophilic matrix,
and after granulation this matrix structure, at any degree of aggregation, is inserted into
a dominant structure formed by polymers or copolymers of hydrophilic type in
anhydrous state or with low level of residue humidity in the presence of one or more
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amphiphilic substances with surfactant properties and subsequently the final mixture is
subjected to compression.
A gastro-protective coating film, capable of preventing the dissolution of the tablet in a
strongly acid environment, is lastly applied to the surface of the compositions.
Upon swallowing, such a mullimatrix coated composition is protected from contact with
gastric and intestinal acids up to reaching an environment with suitable pH, preferably
greater than 5 or 7, where the gastro-protective coating is solubilised and where the
dissolution programme- which will lead it to progressively distribute the dye inserted in
the fom1ulation simultaneously with the advancement within the digestive cavity -
starts.
Furthermore, an object of the present invention is the abovementioned solid
composition for oral administration for diagnostic purpose, preferably in the endoscopic
diagnostic evaluation of inflammatory, ulcerative dysplastic, pre-neoplastic and
neoplastic pathologies of the gastrointestinal tract, more preferably cancerous or
precancerous forms, polyps, pseudopolyps or different inflammatory pathologies of the
gastrointestinal tract.
The composition of the present invention is generally applied for oral administration
during the preparatory stage for the gastro-intestinal endoscopic analysis in a single
solution or in two or more periods of administration. A typical applied administration
pattern provides that the administration of the composition, preferably tablet, occurs at
the end of the preparatory or cleaning stage of the i nlestinal mucosa, gene rail y carried
out through the uptake of purgatives or polyglycolic laxative substances, or during the
preparation procedure, which usually lasts a few hours.
A further administration pattern provides that the administration of the composition of
the invention, preferably a tablet, occurs before the previously mentioned preparatory or
cleaning stage or that said administration partly occurs before and partly during such
preparatory or cleaning step.
Lastly, an object of the present invention is a method for performing endoscopic
evaluations of the gastrointestinal tract, comprising the administration, possibly
repeated, of the abovementioned composition to be preferably carried out within the day
prior to the endoscopy (i.e. within the preceding 24 hours: preparatory stage), such
evaluation being aimed at the diagnosis of inflammatory, ulcerative pre-neoplastic,
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dysplastic or neoplastic pathologies of the gastrointestinal tract, more preferably
cancerous or precancerous forms, polyps, pseudopolyps or different inflammatory
pathologies of the gastrointestinal tract.
According to a preferred embodiment, the administration of the solid composition of the
invention occurs, once or repeated over time, before, simultaneously and/or after the
uptake of the preparation/cleaning composition preceding the endoscopic analysis (for
example, but not exclusively, using the drug available in the market by the name
Moviprep®).
The expression cleaning composition mentioned above is used to indicate the previously
mentioned saline, polyglycolic or laxative solution which is commonly used for
cleaning and washing the intestinal mucosa before the endoscopic analysis, during the
preparatory stage within the preceding 24 hours.
According to a more preferred embodiment of the invention, the solid composition is
taken by the subject intending to carry out the endoscopic evaluation of the colon in two
administrations where one dose is taken before the washing composition as described
above and the subsequent dose is taken after or during the uptake of the washing
composition as described previously. According to such embodiment each dose can be
constituted by one, or more, solid compositions of the invention, preferably one or more
tablets with unitary content corresponding to a fraction of the entire dose to be
administered.
The examples below are meant for clarifying the invention, without entailing any
restrictions whatsoever with respect thereto.
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EXAMPLES
Example 1: Instant-release coated tablet for endoscopv
Description
I Components
Methylene blue
Lecithin
Stearic acid
Mannitol
Microcrystalline cellulose
Hydroxypropyl cellulose
Sodium starch glycolate
Colloidal hydrated silica
UOM
mg
mg
mg
mg
mg
mg
mg
PCT/IB2011/050881
Amt. per
tablet
50.0
3.0
6.0
120.0
50.0
13.0
4.0
2.0
Magnesium stearate mg 2.0
Ll _C_o_a_ti_nug __________________________________________ ~l
Methacrylic acid copolymer type A
mg 12.0
(Eudragit L)
Triethyl citrate mg 1.2
talc mg 5,8
Titanium dioxide mg 3.0
The production process provides for mixing the dye, lecithin, stearic acid and mannitol
up to obtaining a homogeneous mixture. Then microcrystalline cellulose,
hydroxypropyl cellulose, sodium starch glycolate, colloidal silica are added to the
mixture and mixed once again. After adding magnesium stearate, the mixture is
compressed up to obtaining 250 mg tablets. The tablets are then arranged in a tablet
mixer and coated with a methacrylate-based gastro-resistant film and containing triethyl
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citrate as plasticiser in addition to the titanium dioxide dye and talc, an anti-stick
substance. The tablets thus obtained are subjected to a dissolution Lest in an acid
environment for two hours, where they reveal to be resistant to the dyeing substance.
The tablets yield to the dye within a few minutes upon introduction into a neutral pH
environment.
Example 2 : Instant-release coated tablet for endoscopy
Amt. per
Description UOM
tablet
I Components
Methylene blue mg 600.0
Lecithin mg 5.0
Stearic acid mg 10.0
Mannitol mg 340.0
Microcrystalline cellulose mg 123.0
Sodium starch glycolate mg 30.0
Colloidal hydrated silica mg 20.0
Magnesium stearate mg 12.0
~' C_o_a_tt_it~g~--------------------------------------------'
Methacrylic acid copolymer type A
(Eudragit L)
Triethy I citrate
talc
Titanium dioxide
mg
mg
ma 0
mg
30.0
3.0
15.0
7.0
The tablet was obtained through the same process indicated in example 1.
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The dissolution test applied to the tablet of example 2 allowed establishing the
substantial non-dissolution of the tablets in an acid environment with pH at l and the
subsequent dissolution in vitro of the dye when moved to a 6.8 pH.
Example 3: Instant-release coaled tablet for endoscopY
Amt. per
Description ·UOM
tablet
I Components
Methylene blue mg 1200.0
Lecithin mg 10.0
Stearic acid mg 20.0
Mannitol mg 200.0
Hydroxypropyl cellulose Mg 50.0
Microcrystalline cellulose mg 20.0
Sodium starch glycolate mg 50.0
Colloidal hydrated silica mg 30.0
Magnesium stearate mg 20.0
I Coating
Methacrylic acid copolymer type A
mg 40.0
(Eudragit L)
Triethyl citrate mg 4.0
talc mg 20.0
Titanium dioxide mg 9.0
The tablet was obtained through the same process indicated in example 1.
Even in this case, the dissolution test applied to the tablets allowed establishing the
substantial non-dissolution of the tablets in an acid environment with pH at 1 and the
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subsequent dissolution in vitro of the dye when moved to a 6.8 pH, mimetic value of the
intestinal pH.
Example 4: Intestinal release coated tablet for endoscopv
Amt. per
Description UOM
tablet
I Components
Carmine indigo mg 50.0
Lecithin mg 3.0
Stearic acid 6.0
Mannitol mg 120.0
Microcrystalline cellulose mg 40.0
Hydroxypropyl cellulose mg 23.0
Sodium starch glycolate mg 4.0
Colloidal hydrated silica mg 2.0
I Coating
Magnesium stearate mg 2.0
Methacrylic acid copolymer type A
mg 6.0
(Eudragit L)
Methacrylic acid copolymer type B
mg 6.0
(Eudragit S)
Triethyl citrate mg 1.2
talc mg 5,8
Titanium dioxide mg 3.0
The applied process provides for mixing the dye with the lecithin surfactant, stearic
acid, mannitol and half of the required amount of magnesium stearate. After compacting
the mixture followed by granulation, cellulose, sodium starch glycolate, colloidal silica
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16
and the remaining magnesium stearate are added and then, after further mixing, the final
compression is carried out up to obtaining 250 mg tablets. The tablet is then coaled with
a mixture of methacrylic copolymers of type A and B, so as to extend the resistance to
the dissolution in vitro up to a pH ?:.7, characteristic of the ileocecal and colon
environment.
Example 5: Intestinal release coated tablet for endoscopv
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17
The applied process provides for mixing the dye with the lecithin surfactant, stearic acid
and mannitol. After homogeneously dispersing the dye in the mixture, cellulose, sodium
starch glycolate, colloidal silica and the lubricant magnesium stearate arc added and
then, after further mixing, the final compression is carried out up to obtaining 700 mg
tablets. The nuclei are then subjected to coating using a mixture of mcthacrylic
copolymers of type A and B alongside other auxiliary substances: the tablets resist to
the dissolution in vitro in an acid environment and they dissolve at a pH ?:},
characteristic of the ileocecal and colon environment.
Example 6 : Intestinal release coated tablet for endoscopy
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I Coating
Methacrylic acid copolymer type A
mg 6.0
(Eudrauit L)
Methacrylic acid copolymer type B
mg 6.0
(Eudragit S)
Triethyl citrate mg 1.2
talc mg 5,8
Titanium dioxide mg 3.0
The preparation process provides for mixing the dye with lecithin, stearic acid and
microcrystalline cellulose, compaction thereof into wafers followed by dry granulation,
mixing with the remaining components of the nucleus and the final compression to the
weight of 250 mgltab. The coating uses methacrylic derivatives as base and an alcohol
solvent as application phase.
The tablets thus obtained were subjected to dissolution test in vitro, revealing a good
resistance to the acid environment and a progressive transfer of the dye in the neutral
environment with pH at 7.2.
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Example 7: A colon controlled release tablet.
Description.
I Components
Methylene blue
Lecithin
Stearic acid
Methylhydroxypropyl cellulose
Mannitol
Microcrystalline cellulose
talc
Colloidal hydrated silica
Magnesium stearate
I Coating
Methacrylic acid copolymer type A
(Eudragit L)
Methacrylic acid copolymer type A
(Eudragit L)
Triethyl citrate
talc
Titanium dioxide
UOM
mg
mg
mg
mg
mg
mg
mg
ma b
mg
mg
mg
ma b
mg
mg
PCT/IB2011/050881
Amt. per
tablet
200.0
5.0
14.0
!80.0
140.0
140.0
1 0.()
5.0
6.0
16.0
16.0
6,4
15,6
6.0
The composition is obtained through advance mixing and granulation of the dye, the
lecithin as amphiphilic component, the stearic acid as component of the lipophilic
matrix, mannitol and part of the magnesium stearate; after screening the granules
obtained preliminarily, the remaining components and in particular cellulose, capable of
producing the hydrophilic matrix structure are added. The final pharmaceutical form,
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20
obtained by compressing the mixture of powders and granules, weighing about 700 mg,
is subjected to coating with a mixture of copolymers of methacrylic derivatives of type
A and B, supported by a plasticiser, triethyl citrate, by a dye pigment, titanium dioxide,
and by an anti-stick agent, such as talc, using ethylic alcohol as solvent.
The tablet thus obtained revealed in vitro a substantial non-dissolution at acid pH for 2
hours and a progressive dissolution for about 6 hours in a simulated intestinal medium
with 7.2 pH.
Example 8: Colon endoscope experimental test
The same tablet of example 7 was used for carrying out some colon endoscopies in a
human being with extremely positive results. A single tablet was administered to a
subject about 12 hours before carrying out the endoscopy, during the intestinal
preparation step, followed by the uptake of about further 500 ml of water. The time that
elapsed between the uptake of the tablet and the execution of the endoscopy, about 12
hours, was useful to allow the tablet to reach the intestinal colon region and start the
progressive and slow transfer of the dye which, due to the solubilisation in the liquid
present therein, allowed the homogeneous, intense and persistent coloration of the
intestinal mucosa. Actually, after the administration the colon environment revealed
noticeable areas of coloration, allowing a considerable contrast between the pathologic
areas and the normal mucosa which covers the ascending, transverse, descending,
sigmoid and rectal colon regions.
Figures 1-3 show four (4) colon endoscopic images, obtained during the endoscopy of a
patient who has taken the composition of the invention within the 24 hours preceding
the endoscopy. The images clearly show how only some zones of the colon area of the
patient arc coloured, while the others are normal. This shows how after the uptake of the
composition of the invention, the dye highlights solely the pathologic zones of the
examined colon area (see figures 2, 3-4), and not the zones to be considered nonpathologic
(see figure 1).
Thus, this shows the efficiency of the indicated composition when determining the
elective coloration of the pathologic intestinal areas and, by contrast, a non-pathologic
area which, consequently, is free of coloration.
Example 8-bis: Bioavailability study
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21
The same tablet was used for a bioavailability and phannacokinetic study in which the
profile of blood absorption and urinary elimination of the dye administered to healthy
volunteers during a clinical study of Phase I was determined; the phannacokinetic
parameters were compared with those obtained after intravenous administration of a 100
mg dose of dye and they were as follows:
100 mg i.v. vial
200 mg tablets
Tmax
0.10 hrs
16 hrs
C max
2066 ng/ml
1662 ng/ml
AUC (o~t)
11858 nglm1 *h
32941 nglml*h.
The tablets absorption profile is markedly different from that of the vial and it can be
classified as a typical absorption profile of a controlled release formulation, also as
shown by the chart A below.
Example 9 : Colon endoscope experimental test
Using the tablet of example 7, there was carried out an endoscopy test of the colon by
administering two tablets to a subject awaiting colonoscopy, according to the specified
uptake method, i.e. by taking the first dye tablet at the end of the preparatory stage and
the second tablet about 6 hours before carrying out the endoscopy evaluation. The
tablets were administered with abundant water, so as to efficienlly support the in situ
dissolution of the tablets.
Also in this case, the coloration was homogeneous and well marked, allowing carrying
out the test in optimal conditions for diagnostic purposes.
Example 9-bis: Bioavailability study
The same tablet was used for a bioavailability and pharmacokinetic study in which the
profile of blood absorption and urinary elimination of the dye administered to healthy
volunteers during a clinical of Phase I was determined; the pharmacokinetic parameters
measured after the administration of two 200 mg tablets arc the following:
T max C max AUC (lHJ
16 hrs 1636 nglml 38080 nglml *h.
The tablets absorption profile is very similar to the profile obtained with the 200 mg
tablet and, once again, it can be classified as a typical absorption profile of a controlled
release formulation, also as outlined by the chart B below, where the chart with the dot
refers to the 200 mg dose while the one with the triangle refers to the 400 mg one.
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Example I 0: Colon release coated tablet
Description
j Components
Toluidine blue
Carmine indigo
Lecithin
Stearic acid
Mannitol
Methylhydroxypropyl cellulose
Microcrystalline cellulose
Sodium starch glycolate
Colloidal hydrated silica
Magnesium stearate
I Coatiug
Methacrylic acid copolymer type A
(Eudragit L)
Methacrylic acid copolymer type A
(Eudragit S)
Triethyl citrate
talc
Titanium dioxide
22
·.UOM
mo
Mg
mg
mg
mg
mg
mg
mg
mg
mg
mg
mo 0
mg
mg
mg
PCT/IB2011/050881
Amt. per
tablet
400.0
100.0
5.0
10.0
30.0
95.0
10.0
25.0
15.0
10.0
20.0
20.0
4.0
20.0
9.0
The preparation process provides for the formation of a granulate containing toluidine
blue, lecithin, stearic acid, mannitol and part of the magnesium stearate; after
compaction and reduction into granules through screening, the second Carmine indigo
dye, cellulose, sodium starch glycolate, colloidal silica and the remaining magnesium
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23
stearate lubricant arc added. After homogenization, the mixture is compressed to 700
mg and subsequently subjected to coating as described in example 7, using the two
copolymers of the methacrylic acid and the other functional excipicnls.
The tablet thus obtained resists to dissolution in vitro in buffers with pH < 2 and allows
a progressive release of the dye substances in buffers with pH> 7.
Example 11: gastric immediate release coated tablet
Description
I Components
Congo red
Lecithin
Stearic acid
Mannitol
Microcrystalline cellulose
Sodium starch glycolate
Colloidal hydrated silica
Magnesium stearate
I Coating
Methyl hydroxypropyl cellulose
Polyethylene glycol 6000
talc
Titanium dioxide
Amt. per
·•.uoM
tablet
mg 50.0
mg 3.0
mg 6.0
mg 120.0
mg 63.0
mg 4.0
mg 2.0
mg 2.0
mg 12.0
mg 1.2
mg 6.0
mg 3.8
Obtained through the direct compression method, followed by coating in an aqueous
solvent. The tablet dissolves rapidly in vitro within a few minutes, according to the
specifications required by the regulatory authorities for immediate release tablets.
The tablet thus obtained can be used for endoscopic evaluations of the gastroduodenal
sector for highlighting local pathologic growths which can be related to dysplastic or
neoplastic processes still at the initial stage.
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ExamQle 12: Double layer tablet
Arnt. per
Description UOM
tablet
I Layer 1
Congo red mo 0 100.0
Dioctyl sulfosuccinate mg 5.0
Stearic acid mg 10.0
Mannitol mg 100.0
Microcrystalline cellulose mg 100.0
Sodium starch glycolate mg 20.0
Colloidal hydrated silica mg 10.0
Magnesium stearate mu 0 5.0
Layer 2
Methylene blue mg 100.0
Lecithin mg 7.0
Stearic acid mg 10.0
Methylhydroxypropyl cellulose mg 100.0
Mannitol mg 80.0
Microcrystalline cellulose mg 40.0
talc mg 50.0
Colloidal hydrated silica mg 7.0
Magnesium stearate mer 0 6.0
I Coating
Methyl hydroxypropyl cellulose mg 12.0
Polyethylene glycol 6000 mg 1.2
talc mg 6.0
Titanium dioxide mg 3.8
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The process provides for mixing the components of layer 1 and compression thereof,
followed by the compression of a mixture of powders and granules obtained from a
previous compaction of some components of the layer 2, precisely the dye, lecithin,
stearic acid, the microcrystalline cellulose and mannitol with half of the magnesium
stearate, with the remaining co-formulants.
The tablet, weighing about 850 mg, has two differently coloured distinct layers
fommlated for differentially releasing the dye both in the gastric sector and in the
subsequent intestinal sector.

CLAIMS
1. Solid composition for oral administration containing at least one dye in
association with at least one physiologically acceptable excipient, characterised in that it
compnses:
a) a matrix which consists of lipophilic compounds with melting point
below 90°C, and optionally amphiphilic compounds, In which said at
least one dye is at least partly incorporated;
b) an outer matrix which consists of hydrophilic compounds, in which the
lipophilic matrix, and optionally the amphiphilic matrix are dispersed;
c) optionally other physiologically acceptable excipients;
d) optionally a gastro-resistant coating.
2. Composition according to claim 1, wherein said at least one dye is selected
from among methylene blue, congo red, carmine indigo, toluidine blue or mixtures
thereof.
3. Composition according to the preceding claims, wherein said at least one dye
is comprised in an amount ranging between 2 mg and 1000 mg.
4. Composition according to the preceding claims, wherein said at least one dye
is comprised in an amount ranging between 10 mg and 1500 mg
5. Composition according to the preceding claims, wherein said at least one dye is
comprised in an amount ranging between 50 mg and I 200 mg.
6. Composition according to the preceding claims, wherein said at least one dye is
comprised in an amount ranging between 20 mg and 500 mg.
7. Composition according to the preceding claims, wherein said at least one dye is
comprised in an amount equivalent to about 25 mg.
8. Composition according to the preceding claims, wherein said at least one dye is
comprised in an amount equivalent to about 50 mg.
9. Composition according to the preceding claims, in form of tablet or capsule.
10. Composition according to the preceding claims, wherein said lipophilic
compounds are selected from among saturated, unsaturated or hydrogenated long chain
alcohols, saturated, unsaturated or hydrogenated fatty acids, salts thereof, esters or
amides, mono-, eli- or triglycerides of fatty acids, polyethoxylated derivatives thereof,
waxes, ceramides, cholesterol, cholesterol derivatives or mixtures thereof.
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11. Composition according to the preceding claims, wherein said amphiphilic
compounds are selected from among polar lipids of type I or II, ceramides, glycol alkyl
ethers, alkyl sulfate or sulfosuccinate salts or mixtures thereof
12. Composition according to the preceding claims, wherein said polar lipids of type
I or II are selected from among lecithin, phosphatidylcholine, phosphatidylethanolamine
or mixtures thereof, and said glycol alkyl ether is diethylene glycol rnonornelhyl ether.
13. Composition according to the preceding claims, wherein said hydrophilic
compounds are compounds fanning hydrogel, preferably selected from among
polymers or copolymers of the acrylic or methacrylic acid, alkyl vinyl polymers, alkyl
celluloses, hydroxyalkyl celluloses, carboxyalkyl cel!uloses, modified celluloses,
plurisubstituted celluloses, polysaccharides, dextrins, pectins, starches, complex
starches, starch derivatives, alginic acid, synthetic rubber, natural rubber, polyalcohol or
mixtures thereof.
14. Composition according to the preceding claims, wherein said gastro-resistant
coating is selected from among polymers of the acrylic or methacrylic acid, copolymers
of the acrylic or methacrylic acid, cellulose derivatives, hydroxybutyrate-based
polymers, shellac or mixtures thereof.
15. Composition according to any one of the preceding claims for diagnostic
purpose.
16. Composition according to claim 15 for use 111 the endoscopic diagnostic
evaluation of the gastrointestinal tract.
17. Composition according to claim 15 for use m the endoscopic diagnostic
evaluation of inflammatory, ulcerative, dysplastic, pre-neoplastic and neoplastic
pathologies of the gastrointestinal tract.
18. Composition according to claim 15 for use in the endoscopic diagnostic
evaluation of cancerous forms, precancerous forms, polyps, pseudopolyps, or different
inflammatory pathologies of the gastrointestinal tract.
19. Method for the endoscopic diagnostic evaluation of inflammatory, ulcerative,
dysplastic, pre-neoplastic and neoplastic pathologies of the intestine comprising the
administration, within the 24 hours preceding said endoscopic diagnostic evaluation, of
one or more solid compositions as defined in the preceding claims.
20. Method according to claim 19 characterised in that said administration is carried out once
or repeatedly, preferably fractioning the dosage to be administered into two or more uptakes
within the 24 hours preceding the diagnostic evaluation.
21. Method according to claim 19 for the endoscopic diagnostic evaluation of cancerous
or precancerous forms, polyps, pseudopolyps or different inflammatory pathologies of the
gastrointestinal tract.