SOLID ORAL COMPOSITION CONTAINING DYES FOR USE IN ENDOSCOPIC DIAGNOSIS

Abstract

Herein described are solid oral compositions of dyes for use in diagnostic endoscopy, preferably colon endoscopy.

Claims

1. More than two unit dosages of a solid composition containing at least one dye in association with at least one physiologically acceptable excipient which comprises: (a) at least one dye; (b) at least one lipophilic compound; (c) at least one hydrophilic compound; (d) optionally at least one amphiphilic compound; (e) optionally other physiologically acceptable excipients; and (f) optionally a gastro-resistant coating for oral administration for use in an endoscopic procedure for the evaluation of pathologies of the gastrointestinal tract in a human, wherein said more than two unit dosages of said solid composition are to be orally administered to a human according to a fractionated schedule in which a total amount from about 50 mg to about 500 mg, or from about 100 mg to about 400 mg, or from about 100 mg to about 250 mg, or about 200 mg, of said at least one dye is administered to said human in the 48 hour period prior to the endoscopic diagnosis.

2. A method of an endoscopic procedure for the evaluation of pathologies of the gastrointestinal tract in a human, said method comprising: (i) orally administering to the human a bowel cleansing solution and eight unit dosages of a solid composition; and (ii) using an endoscope to evaluate the gastrointestinal tract for said pathologies; wherein the bowel cleansing solution and the eight unit dosages of the solid composition are administered to the human according to a schedule comprising: (a) 3 unit dosages of the solid composition are administered together with or after intake of a second litre of the bowel cleansing solution; (b) 3 unit dosages of the solid composition are administered together with or after intake of a third litre of the bowel cleansing solution; and (c) 2 unit dosages of the solid composition are administered together with or after intake of a fourth litre of the bowel cleansing solution; and wherein each unit dosage of the solid composition comprises: (a) 25 mg of methylene blue; (b) at least one lipophilic compound; (c) at least one hydrophilic compound; (d) optionally at least one amphiphilic compound; (e) optionally other physiologically acceptable excipients; and (f) optionally a gastro-resistant coating.

3. The method of claim 2, wherein the bowel cleansing solution and the eight unit dosages are administered to said human in the 48 hours prior to using an endoscope to evaluate the gastrointestinal tract for said pathologies.

4. The method of claim 2, wherein the dosage units are formulated as tablets.

5. The method of claim 2, wherein the bowel cleansing solution-comprises a PEG-based salt containing solution or laxatives-based solution.

6. The method of claim 2, wherein the bowel cleansing solution comprises a saline or polyglycol solution, or a combination thereof.

7. The method of claim 2, wherein the bowel cleaning solution and said eight unit dosages are administered to said human in the 24 hours prior to use of the endoscope.

8. The method of claim 2, wherein the endoscopic procedure for the evaluation is of inflammatory, ulcerative, pre-neoplastic, dysplastic and/or neoplastic pathologies and/or lesions of the gastrointestinal tract of the human.

9. The method of claim 8, wherein the methylene blue enhances the intestinal mucosal lesion evaluation of cancerous forms, precancerous forms, interval cancers, adenomas, carcinomas, serrated lesions, intraepithelial neoplasias, dysplasias, polyps, pseudopolyps, prepolyps or different inflammatory pathologies and/or lesions sessile, flat, peduncolated shape in the human.

10. The method of claim 8, wherein the endoscopic diagnostic evaluation is of right colon adenomas, right colon polyps and/or interval cancers, in said human.

11. The method of claim 8, wherein the endoscopic procedure for the evaluation is of lesions having a size equal to or less than 5 mm in the human.

12. The method of claim 11, wherein said small lesions are selected from polyps, adenomas and serrated lesions.

13. The method of claim 2, wherein the methylene blue enhances the evaluation of intestinal mucosal lesions in a human suffering from Inflammatory Bowel Disease (IBD), Ulcerative Colitis or Crohn's Disease.

14. The method of claim 2, wherein the methylene blue enhances the evaluation of intestinal mucosal lesions of the right part of the colon of the human.

15. The method of claim 2, wherein administration of the solid composition to said human generates in said human an intraepithelial neoplasias detection outcome with a specificity higher than about 50% or higher than about 80%.

16. A method of staining the mucosa in the gastrointestinal tract in a human comprising: (i) orally administering to the human a bowel cleansing solution and eight unit dosages of a solid composition to the human, wherein the bowel cleansing solution and eight unit dosages are administered to the human according to a schedule comprising. (a) 3 unit dosages of the solid composition are administered together with or after intake of a second litre of the bowel cleansing solution; (b) 3 unit dosages of the solid composition are administered together with or after intake of a third litre of the bowel cleansing solution; and (c) 2 unit dosages of the solid composition are administered together with or after intake of a fourth litre of the bowel cleansing solution; and wherein the unit dosage of the solid composition comprises: (a) 25 mg of methylene blue; (b) at least one lipophilic compound; (c) at least one hydrophilic compound; (d) optionally at least one amphiphilic compound; (e) optionally other physiologically acceptable excipients; and (f) optionally a gastro-resistant coating, whereby the mucosa in the gastrointestinal tract in the human is stained.

17. The method of claim 16, whereby the evaluation of pathologies in the gastrointestinal tract in a human is improved.

18. The method according to claim 17, wherein said pathologies in the gastrointestinal tract in a human are selected from inflammatory pathologies, pre-neoplastic pathologies, dysplastic pathologies, and neoplastic pathologies.

19. The method according to claim 17, wherein said pathologies in the gastrointestinal tract in a human are selected from cancerous pathologies, precancerous pathologies, interval cancers, adenomas, carcinomas, serrated lesions, dysplasias, polyps, pseudopolyps, pre-polyps, hyperplastic lesions, and inflammatory pathologies.

Description

EXAMPLES

Example 1: Controlled-Release Coated Tablet for Endoscopy (Colon)

[0148]

TABLE-US-00001 Description UOM Amt. per tablet Components Carmine indigo mg 50.0 Lecithin mg 5.0 Stearic acid mg 10.0 Mannitol mg 100.0 Lactose mg 50.0 Hydroxyethyl cellulose mg 25.0 Sodium starch glycolate mg 6.0 Colloidal hydrated silica mg 3.0 Magnesium stearate mg 2.0 Coating Methacrylic acid copolymer type A (Eudragit L) mg 6.0 Methacrylic acid copolymer type B (Eudragit S) mg 6.0 Triethyl citrate mg 1.2 talc mg 5.8 Titanium dioxide mg 3.0

[0149] 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, then cellulose, sodium starch glycolate, colloidal silica and the remaining magnesium stearate are added and, after further mixing, the final compression is then carried out to obtain 250 mg tablets. The tablet is then coated with a mixture of methacrylic copolymers of type A and B, so as to extend the resistance to dissolution in vitro up to a pH≥7, characteristic of the ileocecal and colon environment.

Example 2: Controlled-Release Release Coated Tablet for Endoscopy (Colon)

[0150]

TABLE-US-00002 Description UOM Amt. per tablet Components Methylene blue mg 50.0 Lecithin mg 5.0 Stearic acid mg 10.0 Mannitol mg 100.0 Dibasic Sodium phosphate mg 25.0 Hydroxypropyl methylcellulose Mg 35.0 Sodium starch glycolate mg 6.0 Colloidal hydrated silica mg 2.0 Magnesium stearate mg 2.0 Coating Methacrylic acid copolymer type A (Eudragit L) mg 6.0 Methacrylic acid copolymer type B (Eudragit S) mg 6.0 Triethyl citrate mg 1.2 talc mg 5.8 Titanium dioxide mg 3.0

[0151] The preparation process provides for mixing the dye with lecithin, stearic acid and dibasic sodium phosphate, compaction thereof into wafers followed by dry granulation, mixing with the remaining components of the nucleus and the final compression to the weight of 235 mg/tablet. The coating uses methacrylic derivatives as base and an alcohol solvent to facilitate the application phase.

[0152] 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 having a pH at 7.2.

Example 3: Controlled Release Coated Tablet for Endoscopy (Colon)

[0153]

TABLE-US-00003 Description UOM Amt. per tablet Components Methylene blue mg 200.0 Lecithin mg 5.0 Stearic acid mg 14.0 Methylhydroxypropyl cellulose mg 180.0 Mannitol mg 140.0 Microcrystalline cellulose mg 140.0 talc mg 10.0 Colloidal hydrated silica mg 5.0 Magnesium stearate mg 6.0 Coating Methacrylic acid copolymer type A (Eudragit L) mg 16.0 Methacrylic acid copolymer type B (Eudragit S) mg 16.0 Triethyl citrate mg 6.4 talc mg 15.6 Titanium dioxide mg 6.0

[0154] The composition is obtained through advance mixing and granulation of the dye, the lecithin as amphiphilic component, the stearic acid as a 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, obtained by compressing the mixture of powders and granules, and weighing about 720 mg, is subjected to coating with a mixture of copolymers of methacrylic derivatives of type A and B, supported by a plasticiser, i.e., triethyl citrate, by a dye pigment, i.e., titanium dioxide, and by an anti-stick agent, such as talc, using ethyl alcohol as a solvent.

[0155] The tablet thus obtained resists dissolution in vitro in buffers with pH<2 and allows a progressive release of the dye substances in buffers with pH>7 as here below detailed: [0156] Dissolution % after 2 hours in pH 1 dissolution medium: 0% (spec ≤10%) [0157] Dissolution % after 4 hour of pH 7.2 dissolution medium: 27% [0158] Dissolution % after 8 hour of pH 7.2 dissolution medium: 84% (spec >80%)

[0159] The same tablets of this Example 3 have been used for a PK Phase I trial, where 200 and 400 mg single doses have been compared and where the following averaged values of the main PK parameters have been recorded:

[0160] for the 200 mg dose [0161] mean t.sub.lag≥3 hours [0162] mean t.sub.max (hours) 16.10±4.01 [0163] bioavailability compared to injected dose (F.sub.abs %): 139.19±52.0 [0164] mean C.sub.max (ng/ml) 1662.2±501.93 [0165] urine excretion (mean % of the dose)=39.67±19.19 [0166] mean t.sub.1/2 (hours) 20.19±4.68,

[0167] whereas for the 400 mg dose the main parameters recorded have been: [0168] mean t.sub.lag≥3 hours [0169] mean t.sub.max (hours) 17.67±3.60 [0170] mean C.sub.max (ng/ml) 1635.67±729.57 [0171] urine excretion (mean % of the dose)=22.99±14.92 [0172] mean t.sub.1/2 (hours) 17.25±7.43

Example 4: Controlled-Release Coated Tablet for Endoscopy (Colon)

[0173]

TABLE-US-00004 Description UOM Amt. per tablet Tablet Indigo Carmine mg 100.0 Sodium Lauryl sulphate mg 3.0 Stearic acid mg 12.0 Lactose mg 130.0 Microcrystalline cellulose mg 80.0 Sodium starch glycolate mg 10.0 Colloidal hydrated silica mg 12.0 Magnesium stearate mg 3.0 Coating Methacrylic acid copolymer type A mg 10.0 Methacrylic acid copolymer type B mg 10.0 Triethylcitrate mg 8.0 Talc mg 6.0 Titanium dioxide mg 3.8

[0174] 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.

[0175] The tablet, weighing about 250 mg, has two differently coloured distinct layers formulated for differentially releasing the dye both in the gastric sector and in the subsequent intestinal sector.

Example 5: Controlled-Release Coated Tablet for Endoscopy (Colon)

[0176]

TABLE-US-00005 Description UOM Amt. per tablet Methylene blue mg 25.0 Lecithin mg 3.0 Stearic acid mg 10.0 Methylhydroxypropyl cellulose mg 90.0 Mannitol mg 121.0 Microcrystalline cellulose mg 60.0 talc mg 3.0 Colloidal hydrated silica mg 5.0 Magnesium stearate mg 3.0 Coating Methacrylic acid copolymer type A (Eudragit L) mg 8.0 Methacrylic acid copolymer type B (Eudragit S) mg 8.0 Triethyl citrate mg 3.2 talc mg 7.8 Titanium dioxide mg 3.0

[0177] The composition is obtained through ordered mixing of the dye, the lecithin as amphiphilic component, the stearic acid as component of the lipophilic matrix; then the remaining components were added and in particular the celluloses, capable of producing the hydrophilic matrix structure up to completion of the formula. The final pharmaceutical form, obtained by compressing the mixture of powders and granules, unitary weighing of about 320 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-sticking agent, such as talc, using ethyl alcohol or water or mixtures thereof as solvent.

[0178] The tablet thus obtained revealed in vitro a substantial non-dissolution (<10%) at pH 1 for 2 hours and a progressive dissolution in a simulated intestinal medium with pH 7.2 with a release of: [0179] about 10% after 1 hour (with specification limit ≤30%) [0180] about 44% after 4 hours and [0181] more than 90% at the eighth hour (with specification limit ≥80%).

[0182] The tablets have been used also to determine in Human volunteers, subjected to a standard bowel cleansing procedure through the administration of a 4-liters, PEG containing bowel preparation solution (commercially known as Selg® Esse 1000), the PK characteristics of 2 doses of Methylene Blue administered as divided doses individually containing 25 mg of the dye.

[0183] The same tablets have been used for a PK Phase I trial, where 100 and 200 mg single doses have been compared and where the following averaged values of the main PK parameters have been recorded: [0184] for the 100 mg dose [0185] mean t.sub.lag>3 hours [0186] mean t.sub.max (hours) 12.0 (individual values 9-16) [0187] mean C.sub.max (ng/ml) 573.60±175.83 [0188] urine cumulative excretion (mean % of the dose) in 0-60 hours=28.02±11.71 [0189] mean t.sub.1/2 (hours) 13.87±5.09

[0190] whereas for the 200 mg dose the main parameters recorded have been [0191] mean t.sub.lag≥3 hours [0192] mean t.sub.max (hours) 16.0 (individual values 10-24) [0193] mean C.sub.max (ng/ml) 1149.12±261.95 [0194] urine cumulative excretion (mean % of the dose) in 0-60 hours=38.67±15.8 [0195] mean t.sub.1/2 (hours) 15.08±5.85

[0196] In order to optimize the way to administer the tablets as function of the mucosal staining results, a clinical trial has been carried out with the above described tablets, using as discriminating parameter a scoring system (TSC) originally created and composed of a number between 0 and 20, calculated as sum of each individual staining score ranging 0 to 5 (where 0 is not stained at all, 1 is “traces”, i.e. poor dye traces in colonic mucosa, 2 “detectable”, i.e. relevant to a staining of at least 25% of the area, 3 is “acceptable”, i.e. relevant to a staining of at least 50% of the area, 4 is “good”, i.e. relevant to a staining of at least 75% of the area, 5 is “overstained”, i.e. relevant to an overstaining not enabling an endoscopist to see the mucosal surface with the due accuracy in the 100% of the area), measured in the 4 segments of the colonic tract and indicated as right or ascending colon, transverse colon, descending colon and sigma-rectum; this scoring system was used to select the most reliable administration schedule of the dye with the aim of optimizing the tablets administration and the lesions detection possibilities during the colonoscopy procedure.

[0197] So, using the tablets formulated as described, the administration schedules has been changed on small groups of patients and the corresponding staining score has been determined. Since the importance of the colonic mucosal staining is that a well stained aspect should be extended to all the colonic segments, not only focused in a single colonic district, an additional parameter has been taken into account: the NSA or Number of Stained Area with staining score ≥2. With the application of these two parameters (TSC and NSA) the determination of the tablets administration schedule in order to obtain the best conditions for the endoscopist to enhance the detection of all the lesions in the colonic mucosa, has been carried out.

[0198] In the table below the different administration schedules of the two doses tested are reported with the corresponding measured staining score:

[0199] A) for 150 mg dose, [0200] with the administration schedule A including 2 tablets (tbs.) before drinking the bowel prep, 2 tbs. after the first litre (L), 2 tbs. after the second L and the mean staining score was 6.8±4.0 and the mean stained colonic segments (NSA) was 1.3. [0201] with the administration schedule B including 6 tablets (tbs.) before drinking the bowel prep, the mean staining score was 2.3±2. 4 and the mean stained colonic segments (NSA) was 0.4 [0202] with the administration schedule C including 6 tablets (tbs.) at the end of the bowel prep, the mean staining score was 8.1±3. 6 and the mean stained colonic segments (NSA) was 1.5.

[0203] B) for 200 mg dose, [0204] with the administration schedule D including 4 tablets (tbs.) before drinking the bowel prep, 2 tbs. after the first L, 2 tbs. after the second L and the mean staining score was 7.0±5.0 and the mean stained colonic segments (NSA) was 1.3. [0205] with the administration schedule E including 8 tablets (tbs.) at the end of bowel preparation solution the mean staining score was 9.8±4.4 and the mean stained colonic segments (NSA) was 2.3. [0206] with the administration schedule F including 2 tablets (tbs.) before drinking the bowel prep, 2 tbs. after the first L, 2 tbs. after the second L and 2 tbs. at the end of bowel preparation the mean staining score was 9.3±4.1 and the mean stained colonic segments (NSA) was 2.2. [0207] with the administration schedule G including 2 tablets (tbs.) before drinking the bowel prep, 2 tbs. after the first L, 2 tbs. after the second L and 2 tbs. at the end of bowel preparation the mean staining score (TSC) was 10.5±7.8 and the mean stained colonic segments (NSA) was 1.5. [0208] with the administration schedule H including 4 tbs. after the third L, and 4 tbs. at the end of bowel preparation the mean staining score (TSC) was 10.0±3.2 and the mean stained colonic segments (NSA) was 2.1. [0209] with the administration schedule I including 4 tbs. after the second L and 4 tbs. after the third L of bowel preparation the mean staining score (TSC) was 11.4±3.8 and the meanstained colonic segments (NSA) was 2.8. [0210] with the administration schedule J including 2 tablets (tbs.) after the second L 3 tbs: after the third L and 3 tbs. at the end of bowel preparation the mean staining score (TSC) was 11.6±3.5 and the stained colonic segments (NSA) was 2.6.

[0211] Using the same tablets described in Example 5, with a total dose of 200 mg of Methylene blue and an administration schedule of 2 tbs. after the second L, 3 after the third L and 3 at the end of bowel preparation, two Phase II clinical trials have been carried out: A) on 96 completed patients for cancer screening and surveillance, and B) an additional 52 patients belonging to a high risk population, i.e. the patients with long standing Ulcerative Colitis. [0212] A) The cancer screening and surveillance trial had the aim of evaluating the polyp and adenoma detection rate in patients undergoing a full colonoscopy after colonic mucosal staining obtained with Methylene Blue MMX® tablets. Therefore, the primary end-point was to evaluate the polyp detection rate and the adenoma detection rate after colonic mucosal staining

[0213] Other Secondary end-point(s) have been set, precisely: [0214] to classify polyps and adenomas detected after colonic mucosal staining [0215] to evaluate the serrated lesion detection rate. [0216] to evaluate the mucosal staining efficacy of Methylene Blue MMX® tablets [0217] the Bowel cleansing quality was also evaluated according to the validated Boston Bowel Preparation Scale (BBPS). [0218] to collect data about safety and tolerability of Methylene Blue MMX® tablets after administration of a single dose of 200 mg.

[0219] The subjects started the tablets intake in the afternoon before the colonoscopy day and had to drink at least 250 mL of preparation every 15 min, so that the bowel preparation intake could be completed 4 h after.

[0220] Measured trial variables: [0221] frequency of patients with polyps. [0222] Frequency of patients with adenomas. [0223] Number of adenomas in the right colon for each patient. [0224] Number of detected serrated lesions for each patient. [0225] Mucosal staining score for each area; total staining score. [0226] Boston bowel preparation score for bowel cleansing preparation quality. [0227] Time to reach the caecum. [0228] Time to withdrawal from caecum to exit; [0229] adverse events, [0230] vital signs (blood pressure, heart rate, saturation in peripheral blood), body weight.

[0231] The obtained results are here below summarized. [0232] 1) Mucosal abnormalities (polyps, adenomas and serrated lesions) in each colonic region per patient (A) and as total number (B)

TABLE-US-00006 Methylene blue MMX ® tablets Number of Number of Number of Colonic region polyps adenomas serrated lesions (A) All regions 1.8 ± 2.9 1.0 (0-20)  0.9 ± 1.7  0 (0-14) 0.7 ± 1.8  0 (0-10) Right colon 0.6 ± 1.2 0 (0-9) 0.4 ± 1.1 0 (0-8) 0.1 ± 0.4 0 (0-2) Caecum 0.2 ± 0.5 0 (0-3) 0.2 ± 0.4 0 (0-3)   0 ± 0.2 0 (0-2) Ascending 0.3 ± 0.6 0 (0-3) 0.2 ± 0.6 0 (0-3) 0.1 ± 0.3 0 (0-2) colon Hepatic flexure 0.2 ± 0.6 0 (0-5) 0.1 ± 0.5 0 (0-4)   0 ± 0.1 0 (0-1) Transverse 0.1 ± 0.4 0 (0-2) 0.1 ± 0.3 0 (0-1)   0 ± 0.2 0 (0-1) colon Splenic flexure 0.1 ± 0.3 0 (0-2) 0.1 ± 0.3 0 (0-2) 0 ± 0 0 (0-0) Descending 0.1 ± 0.3 0 (0-1) 0.1 ± 0.2 0 (0-1)   0 ± 0.2 0 (0-1) colon Sigmoid 0.4 ± 0.8 0 (0-4) 0.1 ± 0.4 0 (0-2) 0.2 ± 0.6 0 (0-3) Rectum 0.5 ± 1.6  0 (0-10) 0.1 ± 0.6 0 (0-5) 0.4 ± 1.3 0 (0-9) (B) All regions 61 (63.5) 45 (46.9) 26 (27.1) Right colon 32 (33.3) 24 (25.0) 9 (9.4) Caecum 14 (14.6) 13 (13.5) 2 (2.1) Ascending 16 (16.7) 10 (10.4) 5 (5.2) colon Hepatic flexure 9 (9.4) 7 (7.3) 2 (2.1) Transverse 12 (12.5) 8 (8.3) 4 (4.2) colon Splenic flexure 6 (6.3) 5 (5.2) 0 (0.0) Descending 7 (7.3) 4 (4.2) 3 (3.1) colon Sigmoid 21 (21.9) 12 (12.5) 8 (8.3) Rectum 19 (19.8) 9 (9.4) 12 (12.5)

[0233] All endoscopic findings were classified by the histopathologist. The detected lesions were predominantly low grade tubular adenomas, hyperplastic serrated lesions, low grade serrated adenomas, low grade tubular-villous adenomas but also high grade adenomas with carcinoma in situ, including tubular-villous, villous and tubular lesions. The mucosal staining efficacy of Methylene Blue MMX® tablets was on average “acceptable” with the 50% of the mucosa stained in all 4 examined colonic regions. Bowel cleansing quality was on average “good” according to the total BBPS score.

Conclusions:

[0234] The polyp detection rate and the adenoma detection rate/patient in the whole colon were on average 1.8±2.9 detected polyps and 0.9±1.7 detected adenomas. The polyp detection rate ranged from 0 to 20 polyps per subject and was higher in the rectum with a maximum of 10 polyps and in the right colon with a maximum of 9 lesions. The adenoma detection rate ranged from 0 to 14 adenomas per subject and was higher in the rectum with a maximum of 5 adenomas. In the right colon, the maximum detection rate was 8 detected adenomas. Serrated lesions ranged from 0 to 10, with the highest prevalence in the rectum with a maximum of 9 lesions.

[0235] As summarized in the following table. pPolyps were detected at a frequency of 64%, adenomas at a frequency of 47% and serrated lesions at a frequency of 27.1% (9% of subjects in the right colon, considered at the same severity level than adenomas).

TABLE-US-00007 Number of patients Number of patients Number of patients with polyps (%) with adenomas (%) with serrated (%) 61 (63.5) 45 (46.9) 26 (27.1)

[0236] There was a good consistency between the pit pattern scores and histological classification.

[0237] The most frequently affected region for polyps was sigmoid and rectum (21.9% and 19.8% respectively) and serrated lesions most frequently in the rectum (12.5%). Considering the 3 areas right, transverse and descending colon, the transverse colon is that with the lowest detection rate, followed by right and descending colon.

[0238] The analysis was performed also by subdividing the intraepithelial neoplasiae by size. The rate of detection by lesion size is summarised in the following table. The number of detected polyps, adenomas and serrated lesions <5mm; mean (±SD) and median (range) are reported.

TABLE-US-00008 Methylene blue MMX ® tablets Number of Number of Number of Lesion size polyps adenomas serrated lesions ≤5 mm 1.3 ± 2.3 0.5 ± 1.1 0.6 ± 1.7

[0239] Smaller lesions (≤5 mm) were predominant in frequency, and that is remarkable inasmuch that the conventional white light colonoscopy, such smaller lesions are the most difficult to detect. Polyps ≤5 mm had a maximum number of 15 detected abnormalities. The maximum number of detected adenomas ≤5 mm was 9 and 10 for the serrated lesions ≤5 mm.

[0240] Proportion of subjects with detected polyps by size, with detected adenomas and with detected serrated lesions presented also in the following summary table. The proportion of subjects with detected polyps, adenomas and serrated lesions by colonic region; number (%) of subjects is reported

TABLE-US-00009 Methylene blue MMX ® tablets Subjects Subjects Subjects with at least with at least with at least one serrated Popu- Lesion one polyp one adenoma lesion lation size n (%) n (%) n (%) FAS ≤5 mm 50 (52.1) 30 (31.3) 23 (24.0) (N = 96) 6-9 mm 12 (12.5) 10 (10.4) 3 (3.1) ≥10 mm  24 (25.0) 22 (22.9) 3 (3.1)

Conclusions:

[0241] Efficacy of Methylene Blue MMX® 25 mg modified release tablets was investigated and proved in the detection of the mucosal lesions in all the colonic districts, particularly with the lesions <5mm. A large proportion of patients, compared to data in the literature, has been found affected by the presence of polyps and adenomas, particularly in the sigmoid-rectum district and also in the right colon. [0242] B) The efficacy of Methylene Blue MMX® 25 mg modified release tablets was investigated in patients with ulcerative colitis with a diagnosis of ≥8 years and colitis activity index<8, This population was chosen because patients with long standing ulcerative colitis have a significantly higher risk for the development of colitis associated colorectal cancers.

[0243] The intraepithelial neoplasia detection rate was 16% (8 out of 50 subjects belonging to PP population) with a total of 10 intraepithelial neoplasiae detected in the 8 subjects. Intraepithelial neoplasiae were most frequently found in the rectum-sigma segment (RES), followed by descending colon (DC) and tansverse colon (TC) at the same frequency, and finally by the ascending colon (AC). The number of intraepithelial neoplasiae/subject was 0.2±0.5.

[0244] As summarized below, false positive findings represented 8% (4 out of 50 subjects), whilst the false negative findings were 6% (3 out of 50). The method had a sensitivity greater than 50% (precisely 57.1%) and a specificity greater that 90% (precisely 90.7%.)

[0245] Study results are consistent with the higher range of the literature data obtained with the chromo-endoscopy technology a spray of the dye instead of the oral administration of the dye during bowel preparation as disclosed herein. The dye spray technology was able to dramatically reduce the time of examination compared to the random biopsies: in the cited spray chromo-endoscopy trial, intraepithelial neoplasiae were detected at a rate of 15.48% in the same population, with a solution of 0.1% methylene blue sprayed using a catheter.

[0246] Detection rate of intraepithelial neoplasiae and true and false positive and negative findings analysis population (N=52).

TABLE-US-00010 Proportion of subjects with True False True False intraepithelial positive positive negative negative neoplasiae findings findings findings findings 8 (15.4) 4 (7.7) 4 (7.7) 41 (78.8) 3 (5.8)

[0247] The mucosal staining efficacy of Methylene Blue MMX® tablets was confirmed on average “acceptable” with 50% of the mucosa stained in all 4 examined colonic regions, with the best stained colonic segment resulting in the ascending colon, the region where it's more difficult to find the dysplastic lesions. The majority of subjects had NSA in all 4 regions. Bowel cleansing quality was on average “good” according to the total BBPS score.

[0248] Two images of colon endoscopy are below reported to also better clarify the invention.

[0249] FIG. 1 shows the contrast enhancing efficacy of the dye according to the present invention in perceiving the deep mucosal tissue structure, with the foci of the glands well defined and darkened in a pre-polyp alteration of the colonic mucosa.

[0250] FIG. 2 shows the semi-continuous blue line defines exactly the borders of the colonic flat lesion that the endoscopist has to take out, allowing a better resolution of the lesion intervention and extraction. The tissue definition is absolutely enhanced owing to the orally administered dye as disclosed herein. With the conventional spraying techniques, the same performance cannot be obtained since little time is available between spray and observation (seconds or a couple of minutes).