US 2011 O15893 OA1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2011/0158930 A1 Hirata et al. (43) Pub. Date: Jun. 30, 2011
(54) METHOD FOR TREATMENT OF RRTABLE Related U.S. Application Data BOWEL, SYNDROME (60) Provisional application No. 61/190.559, filed on Aug. 28, 2008. (75) Inventors: Takuya Hirata, Tokyo (JP); Toshiyuki Funatsu, Tokyo (JP); Publication Classification Yoshihiro Keto, Tokyo (JP); (51) Int. Cl. Shinobu Akuzawa, Tokyo (JP) A63L/78 (2006.01) C07D 403/06 (2006.01) (73) Assignee: ASTELLAS PHARMA INC., A6IPI/00 (2006.01) Tokyo (JP) A6IPL/T2 (2006.01) (52) U.S. Cl...... 424/78.01: 548/302.7 (21) Appl. No.: 13/061,384 (57) ABSTRACT A method for treatment of a patient suffering from irritable (22) PCT Fled: Aug. 27, 2009 bowel syndrome with diarrhea or mixed irritable bowel syn drome, which comprises administering to the patient a thera (86) PCT NO.: PCT/UP2009/064904 peutically effective amount of ramosetron or a pharmaceuti cally acceptable salt thereof in combination with a S371 (c)(1), therapeutically effective amount of polycarbophil or a phar (2), (4) Date: Feb. 28, 2011 maceutically acceptable salt thereof. Patent Application Publication Jun. 30, 2011 Sheet 1 of 2 US 2011/O1589.30 A1
Fig.1
s 120 aw *e 100 aS$ 100 80 isd 80 v s 60 se sk sk 60 40 6 40 se g 20 20 - a k es O -- - - - , , , g- O W 0. 0.3 1 100 300 000 0.3 g/kg 300 mg/kg 0.3 g/kg + s S. 300 mg/kg Ramosetron Polycarbophi s Š w gkg) (mg/kg) s? s S s cSSk 2 s SS ; SS {r q'sS. &
Fig.2
A B it; 120 s 100 s s
OO 80 80 - 60 60 3 40 ce: 40 20 is 20 ra r 0 8.- 2 0 OW 1. 3. 100 100 300 OO W 30 g/kg 300 mg/kg 30 g/kg + r------KY S. 300 mg/kg Ramosetron Polycarbophil s S gkg) (mg/kg) ass g s S. c S. k q'ss & Fig. 3 6 250 5 200 g sk ss 150 - 3 100 – 50 H O OW 10 30 OO 100 30 OOO Ramosetron Polycarbophil (ug/kg) (mg/kg) Patent Application Publication Jun. 30, 2011 Sheet 2 of 2 US 2011/O1589.30 A1
Fig.4
250
2OO 150 1OO 50 O ---- DW 3OO 1000 DW 300 1000 Polycarbophil Polycarbophil (mg/kg) (mg/kg) Ramosetron 100 g/kg
Fig.5
250
200 - 150 100 50
OW 3OO 1000 DW 3OO 1000 Polycarbophil Polycarbophil (mg/kg) (mg/kg) Ramosetron 100 uglkg US 2011/0158930 A1 Jun. 30, 2011
METHOD FOR TREATMENT OF RRTABLE 0007 Patent Reference 3: U.S. Pat. No. 3,297,664 BOWEL, SYNDROME 0008 Patent Reference 4: WO 2004/062623 TECHNICAL FIELD 0009 Patent Reference 5: WO 2008/096777 0001. The present invention relates to a treatment method of using ramosetron or a pharmaceutically acceptable salt Non-Patent References thereof in combination with polycarbophil or a pharmaceuti (0010. Non-Patent Reference 1: Gastroenterology, 2006; cally acceptable salt thereof for treatment of a patient suffer 130: p. 1480-1491 ing from irritable bowel syndrome with diarrhea or mixed 0011 Non-Patent Reference 2: Japanese Journal of Phar irritable bowel syndrome. macology, 2002; 89: p. 133-141 BACKGROUND ART 0012 Non-Patent Reference 3: Neurogastroenterology 0002 Irritable bowel syndrome (IBS) is a functional gas and Motility, 2008; 20(5): p. 557-565 trointestinal disorder with which the patient experiences chronic disturbance of bowel habit (constipation or diarrhea) DISCLOSURE OF THE INVENTION and abdominal pain/discomfort without any organic disease 0013 The present inventors have assiduously studied for in the gastrointestinal tract. According to the diagnostic cri the purpose of creating a more excellent treatment method for teria for IBS (Rome III Criteria) published in 2006, IBS is patients suffering from IBS with diarrhea or mixed IBS. categorized as IBS with constipation, IBS with diarrhea, 0014. As a result, the inventors have confirmed an excel mixed IBS or unclassified IBS (for example, see Non-Patent lent synergistic effect of ramosetron in combined administra Reference 1). It is reported that the prevalence of IBS to the tion with polycarbophil both in the pharmacological efficacy total population reaches 10% to 20% and IBS patients and the side effect thereof, and have completed the present account for about 20 to 40% of outpatients in gastrointestinal invention. medicine department, and it is considered that IBS is a gas 0015 Using a restraint stress induced rat diarrheal model trointestinal disorder recognized at high frequency. Further, and a 5-HT induced mouse diarrheal model, the inventors the IBS symptom greatly worsens the quality of life and the investigated the Synergistic effect of ramosetron and polycar work productivity of the patient, which is being a serious bophiland have found that the combined use of the two drugs Social problem. At present, for the pharmacological treatment significantly enhances the respective inhibitory effects for of IBS, mainly used is an antispastic drug Such as anticholin diarrhea. Further, the inventors have found that the repeated ergic drug, an antidiarrheal drug such as opioid receptorago combined administration of ramosetron and polycarbophil nist, or a bulking-agent or a probiotics for regulating the attenuates the inhibitory effect of ramosetron on spontaneous enteric environment. defecation in mice. On the other hand, it is reported that a 0003 Ramosetron is a serotonin (5-HT) receptor antago high-molecular polymer Such as colestimide or sevelamer nist having a chemical name of (-)-(R)-5-(1-methyl-1H hydrochloride adsorbs the other low-molecular compound indol-3-yl)carbonyl-4,5,6,7-tetrahydro-1H-benzimidazole administered simultaneously therewith and attenuates the (for example, see Patent Reference 1). Heretofore, ramo absorption and the effect of the compound (Kayset al., Am. J. setron has been prescribed for an adult patient with the gas Kidney Dis. 2003:42: 1253-1259;Yamada et al., Pharmacol trointestinal symptom Such as vomiting caused by antine ogy and Therapy, 2001; 29: 37–44). However, oral adminis oplastic agents, in a mode of oral administration at a dose of tration of a prepared mixed solution of ramosetron and poly 0.1 mg once a day or intravenous injection at a dose of 0.3 mg carbophil has been confirmed to possess the antidiarrheal once a day. Recently, it has been confirmed that ramosetron effect, but on the contrary, rather provides significant effect hydrochloride at an extremely low daily dose of 0.001 to 0.05 enhancement; and therefore, it is considered that the possi mg is also effective for treatment of IBS with diarrhea (for bility that polycarbophil may adsorb ramosetron to attenuate example, see Patent Reference 2). At present, there are only a the effect thereof would be low. few reports showing the efficacy of the combined use of 5-HT, receptor antagonist with any other agent for IBS, such SUMMARY OF THE INVENTION as a noradrenaline reuptake inhibitor and a mast-cell degranu lation inhibitor (for example, see Patent References 4 and 5), 0016 Specifically, the invention relates to the following: while it is still unclear whether the concomitant administra 0017 (1) A method for treatment of a patient suffering tion of ramosetron with the existing agents for IBS described from irritable bowel syndrome with diarrhea or mixed irri above show the beneficial synergistic effect. table bowel syndrome, which comprises administering to the 0004. On the other hand, polycarbophil is a polyacrylic patient a therapeutically effective amount of ramosetron or a acid crosslinked with divinylglycol, and is a high-molecular pharmaceutically acceptable salt thereof in combination with polymer having high water absorbability (for example, see atherapeutically effective amount of polycarbophil or a phar Patent Reference 3). Polycarbophil is used as an antidiarrheal maceutically acceptable salt thereof; drug or for treatment of altered bowel habit in IBS. Specifi 0018 (2 The method of 1 comprising administering to cally, it is considered that, owing to its water absorbing effect, the patient 0.001 to 0.05 mg of ramosetron hydrochloride as polycarbophil could absorb water in an intestinal lumen a daily dose or an equivalent molar amount of ramosetron or increased by stress or the like, therefore normalizing the stool other pharmaceutically acceptable salt thereof, and condition to prevent diarrhea (for example, see Non-Patent 0019. 3. The method of 1 or 2 comprising administer References 2 and 3). ing to the patient 1.0 to 5.0 g of polycarbophil calcium as a daily dose or an equivalent molar amount of polycarbophil or PRIOR ART REFERENCES other pharmaceutically acceptable salt thereof. Patent References 0020. The invention also relates to the following: 0005 Patent Reference 1: EP 381422 0021 4Use of ramosetron or a pharmaceutically accept 0006 Patent Reference 2: US 2005/0192329 able salt thereof for the manufacture of a medicament for the US 2011/0158930 A1 Jun. 30, 2011
treatment of irritable bowel syndrome with diarrhea or mixed each group with n=12, in which *P<0.05/3, **P<0.01/3 to irritable bowel syndrome in combination with polycarbophil distilled water administration group (Fisher's exact test). B or a pharmaceutically acceptable salt thereof, and shows the incidence of diarrhea in each group with n=24, in 0022 5. Use of ramosetron hydrochloride in a daily dose which *P<0.05, **P<0.01 to distilled water administration of 0.001 to 0.05 mg, or an equivalent molar amount of ramo group, #P-0.05, #P-0.01 to ramosetron or polycarbophil setron or other pharmaceutically acceptable salt thereof, for administration group (Fisher's exact test). the manufacture of a medicament for the treatment of irritable 0034 FIG. 3 shows a graph indicating the influence of bowel syndrome with diarrhea or mixed irritable bowel syn single-administration of ramosetron or polycarbophil alone drome in combination with polycarbophil or a pharmaceuti on spontaneous defecation in mice, in which the data indicate cally acceptable salt thereof. the mean value:SEM in each group with n=10, and *P<0.05 0023 The invention further relates to the following: to distilled water administration group (Dunnett's test). 0024 6 A pharmaceutical composition comprising 0035 FIG. 4 shows a graph indicating the influence of ramosetron or a pharmaceutically acceptable salt thereof and single administration of ramosetron, polycarbophil or their polycarbophil or a pharmaceutically acceptable salt thereof; combination on spontaneous defecation in mice, in which the 0025 7. The pharmaceutical composition of 6 compris data indicate the mean value:SEM in each group with n=10 inga) 0.001 to 0.05 mg of ramosetron hydrochloride as a daily to 14, and #P-0.05 to distilled water alone administration dose or an equivalent molar amount of ramosetron or other group (Student's t-test). pharmaceutically acceptable salt thereof, and b) 1.0 to 5.0 g of 0036 FIG. 5 shows a graph indicating the influence of polycarbophil calcium as a daily dose or an equivalent molar repeated administration of ramosetron, polycarbophil or their amount of polycarbophil or other pharmaceutically accept combination on spontaneous defecation in mice, in which the able salt thereof, and data indicate the mean value:SEM in each group with n=10. 0026 8 The pharmaceutical composition of 6 or 7 for iP<0.05 to distilled water alone administration group (Stu use for the treatment of irritable bowel syndrome with diar dent's t-test), and *P-0.05 to distilled water+ramosetron rhea or mixed irritable bowel syndrome. administration group (Dunnett's test (multigroup)). 0027. The invention further relates to the following: 0028 9 An enhancer of the effect of polycarbophil or a BEST MODE FOR CARRYING OUT THE pharmaceutically acceptable salt thereof for improving the INVENTION diarrhea symptom of irritable bowel syndrome, which com prises ramosetron or a pharmaceutically acceptable salt 0037. The invention is described in more detail here thereof, and inunder. 0029 [10] The enhancer of 9), comprising 0.001 to 0.05 0038 Ramosetron and its pharmaceutically acceptable mg of ramosetron hydrochloride as a daily dose or an equiva salt may be readily obtained by the method described in lent molar amount of ramosetron or other pharmaceutically Patent Reference 1 or according to it. acceptable salt thereof. 0039. The pharmaceutically acceptable salt of ramosetron concretely includes mineral acid salts with hydrochloric acid, ADVANTAGE OF THE INVENTION Sulfuric acid, phosphoric acid, hydrobromic acid, etc.; Salts with organic acids Such as acetic acid, oxalic acid. Succinic 0030. According to the invention, there is provided a more acid, citric acid, maleic acid, malic acid, fumaric acid, tartaric excellent treatment method for patients suffering from IBS acid, methanesulfonic acid, etc.; Salts with acidic amines Such with diarrhea or mixed IBS. as glutamic acid, aspartic acid, etc. Above all, preferred is 0031 Specifically, combined use of ramosetron or a phar commercially-available ramosetron hydrochloride. maceutically acceptable salt thereof and polycarbophil or a pharmaceutically acceptable salt thereof provides a beneficial 0040 Polycarbophil and its pharmaceutically acceptable effect on IBS symptoms even to the patients who could not salt may be readily obtained by the method described in obtain sufficient symptomatic improvement by the individual Patent Reference 3 or according to it. drugs alone. Further, the combined use with polycarbophil or 0041. The pharmaceutically acceptable salt of polycarbo a pharmaceutically acceptable salt thereof can inhibit the phil concretely includes alkali metal salts with sodium, potas sium, etc.; alkaline earth metal salts with calcium, magne occurrence of constipation as the side effect in relation to the sium, etc.; aluminium salts; ammonium salts; salts with administration of ramosetron or a pharmaceutically accept organic amines such as benzathine, choline, diethanolamine, able salt thereof. ethylenediamine, meglumine, diethylamine, piperazine, tromethamine, procaine, etc. Above all, preferred is commer BRIEF DESCRIPTION OF THE DRAWINGS cially-available polycarbophil calcium. 0032 FIG. 1 includes graphs showing the effect of ramo 0042 Ramosetron or its pharmaceutically acceptable salt setron, polycarbophil and their combination for restraint and polycarbophil or its pharmaceutically acceptable salt stress-induced diarrhea in rats. A shows the incidence of may be, either separately or simultaneously, orally adminis diarrhea in each group with n=12, in which *P-0.05/3, tered as a pharmaceutical composition thereof with an **P<0.01/3 to distilled water administration group (Fisher's organic or inorganic carrier, vehicle and other additive Suit exact test). B shows the incidence of diarrhea in each group able to oral administration. In case where the active ingredi with n=24, in which *P<0.05, **P<0.01 to distilled water ents are separately formulated in different preparations, those administration group, #P-0.05, #P-0.01 to ramosetron or different preparations may be administered to a patient sepa polycarbophil administration group (Fisher's exact test). rately or simultaneously or at time intervals. 0033 FIG. 2 includes graphs showing the effect of ramo 0043. The pharmaceutical composition concretely setron, polycarbophil and their combination for 5-HT-in includes tablets, powders, granules, microspheres, capsules, duced diarrhea in mice. A shows the incidence of diarrhea in pills, syrups, emulsions, Suspension, etc. US 2011/0158930 A1 Jun. 30, 2011
0044. In the oral preparation, the active ingredient is under the acidic condition; and for example, the amount is mixed with at least one inert diluent, for example, lactose, from 1 to 80% by weight of polycarbophil calcium in the mannitol, glucose, microcrystalline cellulose, starch, polyvi preparation. nyl pyrrolidone, magnesium metasilicate aluminate. The 0048. The dose (daily dose) of ramosetron or its pharma composition may contain, according to an ordinary method, ceutically acceptable salt and polycarbophil or its pharma any other additive than the inert diluent, for example, a binder ceutically acceptable salt may be suitably determined case by Such as hydroxypropyl cellulose, hydroxypropylmethyl cel case, depending on the symptom of the disease, the age, the lulose; a lubricant such as magnesium Stearate, calcium Stear race and the sex of the object intended for administration, etc. ate, polyethylene glycol, starch, talc, a disintegrant Such as The daily dose of a drug as referred to herein means the total amount of the drug to be administered within 24 hours, and calcium glycolate cellulose; a stabilizer Such as lactose; a the total amount may be administered in a mode of single dose dissolution promoter Such as glutamic acid, aspartic acid; a (one time administration a day) or multiple dose (two or more plasticizer Such as Tween 80, triacetin; a colorant such as times administration within 24 hours, and the total of the titanium oxide, iron sesquioxide. Tablets or pills may be multiple dose is within the range of the daily dose as referred optionally coated with a Sugar coating film or a film of a to herein). gastric or enteric Substance Such as Sucrose, gelatin, agar, 0049 Ramosetron hydrochloride may be administered to pectin, hydroxypropyl cellulose, hydroxypropylmethyl cel a patient with IBS generally in a daily dose of about from lulose phthalate, etc. 0.001 to 0.05 mg/adult in a mode of ordinary oral adminis 0045. The pharmaceutical composition may be an oral tration, most preferably in a daily dose of from 0.0025 to 0.01 disintegrable tablet. For example, oral disintegrable tables mg; and this is orally administered once a day. Polycarbophil may be prepared according to U.S. Pat. No. 5,466,464, U.S. calcium may be administered generally in a daily dose of Pat. No. 5,576,014, U.S. Pat. No. 6,589,554, WO2003/ about from 1 to 8 g/adult in a mode of ordinary oral admin 009831, WO2002/092057, etc. istration, most preferably in a daily dose of from 1.5 to 3.0 g; 0046 Ramosetron is used in an extremely low dose, and is and this is orally administered along with water, once or as therefore especially preferably in the form of a preparation divided in two to four times a day. specifically stabilized to temperature and humidity. For 0050. Accordingly, for example, polycarbophil calcium example, as described in US2005/0026981 A1, a specific may be administered three times a day after every meal each compound having a carbonyl group may be added thereto to in a dose of from 0.5 to 1.0 g, and in one time of those, stabilize ramosetron to temperature and humidity. The spe ramosetron hydrochloride may be administered as combined cific compound having a carbonyl group includes, concretely, in a dose of from 0.0025 to 0.01 mg. aliphatic carboxylic acids (in detail, Saturated or unsaturated, 0051 Various treatment plans may be employed. Some linear or branched aliphatic mono-, di or tri-carboxylic acid, patients may be treated occasionally. For example, ramo especially aliphatic carboxylic acids having from 3 to 36 setron or its pharmaceutically acceptable salt and polycarbo carbon atoms) or their esters; hydroxycarboxylic acids (in phil or its pharmaceutically acceptable salt are administered detail, Saturated or unsaturated, linear or branched aliphatic each in the above-mentioned daily dose to the patient who has hydroxy-mono-, di- or tri-carboxylic acids, especially developed a Sudden onset of IBS symptoms, during a suffi hydroxycarboxylic acids having from 3 to 36 carbon atoms) cient period of time for relieving the symptom of IBS. For or their esters; acidic amino acids, enol acids, aromatic car example, ramosetron or its pharmaceutically acceptable salt boxyl compounds (in detail, aromatic mono-, di or tri-car and polycarbophil or its pharmaceutically acceptable salt are boxylic acids optionally Substituted with an alkyl group hav administered to the patient each in the above-mentioned daily ing 1 to 4 carbon atoms or a hydroxy group, especially dose for a period of 7 days, 14 days, 21 days, 28 days, 6 aromatic carboxylic acids having from 7 to 20 carbonatoms) weeks, 8 weeks, 12 weeks, 16 weeks or longer until the or their esters, and polymer Substances having a carboxyl symptom of IBS could disappear. Some other patients may be group. One or more these compounds may be used here either treated continuously for a longer period of time. For example, singly or as combined. Not specifically defined, the amount to the above-mentioned daily dose of ramosetron or its pharma be used of the compound to stabilize ramosetron or its phar ceutically acceptable salt is preventively administered occa maceutically acceptable salt may be any one capable of sionally or continuously during a long period of time (at least attaining the stabilization. For example, in the preparation, for 6, 9, 12 or 15 months) for the purpose of evading or the amount may be from 0.01 to 90% by weight, preferably inhibiting the onset of IBS. Further, any other treatment plans from 0.01 to 50% by weight, more preferably from 0.1 to 10% may also be employable. by weight in consideration of the producibility of the prepa 0.052 According to Rome III diagnostic criteria, IBS is ration. categorized as IBS with constipation, IBS with diarrhea, mixed IBS or unclassified IBS (Longstreth et al., Gastroen 0047 Polycarbophilis highly sticky, and therefore, in case terology, 2006: 130: 1480-1491); combined administration of where it is orally administered as a tablet, preferably the tablet ramosetron or its pharmaceutically acceptable salt and poly can be fully disintegrated in the stomachand can be uniformly carbophil or its pharmaceutically acceptable salt is consid dispersed therein. For example, as a chewable tablet, the preparation may be chewed by a patient in its mouth and the ered effective for the diarrheal symptom of patients suffering thus-disintegrated preparation may be introduced into the from IBS with diarrhea and mixed IBS. stomach. In addition, as described in EP 488139, a cellulose derivative may be incorporated in a preparation of polycar EXAMPLES bophil to thereby make the preparation disintegrable under 0053. The invention is described in more detail with ref the acidic condition in a stomach. Not specifically defined, erence to the following Examples and Experimental the amount to be added of the cellulose derivative may be any Examples; however, the invention is not limited to these one capable attaining the disintegration of the preparation Examples, etc. US 2011/0158930 A1 Jun. 30, 2011
Example 1 Example 3 0.058 0054
Polycarbophil calcium 62.5 parts Ramosetron hydrochloride 0.0008 parts Carboxymethyl cellulose 1.25 parts Crystalline cellulose quantum libet Mannitol 89 parts Magnesium stearate 0.6 parts Citric anhydride 0.1 parts Hydroxypropylmethyl cellulose 2 parts Maltose 10 parts Macrogol 6000 0.5 parts Red iron sesquioxide 1 part Titanium oxide 0.5 parts Magnesium Stearate 1 part 0059 A part (about /2 of the total amount) of carboxym ethyl cellulose was added to polycarbophil calcium and 0055 Ten parts of maltose, 0.0008 parts of ramosetron mixed at room temperature, then granulated with water in an hydrochloride, 0.1 parts of citric anhydride and 1 part of red amount of 5% by weight of polycarbophil, and dried at 60° C. iron sesquioxide were suspended in 67 parts of water with for about 10 hours. The granulated product was regulated for stirring with a magnetic stirrer to prepare a spray liquid (con particle size through a 18-mesh sieve, and then the remaining centration, 15% by weight). Next, 89 parts of mannitol was carboxymethylcellulose and crystalline cellulose were added fed into a fluidized bed granulator (FLOW COATER, by thereto, and further magnesium Stearate was added thereto to Freund), and the above spray liquid was sprayed onto it at a give a powder to be tabletted. This was tabletted into tablets spraying speed of 10 g/min for fluidized bed granulation. each containing 625 mg of polycarbophil calcium in one After the granulation, the granulated product was dried at a tablet. The tables were coated with a film of hydroxypropyl suction temperature of 40°C. for 5 minutes, and then mixed methyl cellulose, Macrogol 6000 and titanium oxide to be with 1 part of magnesium Stearate. Using a rotary tabletter, film-coated tablets. the mixed powder was tabletted into tablets each weight 120 Experimental Examples mg and having an initial hardness of about 1 kp. This was I. Test Method: stored at 25°C. and at a relative humidity of 75% for 18 hours, (1) Material and Method: and then stored at 30°C. and at a relative humidity of 40% for 0060. The animals were housed under free drinking and 4 hours, thereby giving oral disintegrable tablets. free feeding condition in a temperature (22+2° C.) and humidity (55-5%)-controlled animal room in a dark-light cycle of 12 hours. All the animal experiments were carried out Example 2 under the approval by Animal Ethical Committee of Astellas Pharma. 0056 0061 Ramosetron hydrochloride, polycarbophil calcium (Astellas Pharma, Inc.) and 5-HT creatinine sulfate (Wako Pure Chemical Industries Ltd.) were used. Ramosetron was Ramosetron hydrochloride 0.0008 parts dissolved and diluted with distilled water. Polycarbophil was Mannitol 88 parts prepared by decalcification of polycarbophil calcium accord Maltose 10 parts ing to a reported method (Yamada et al., Iyakuhin Kenkyu Yellow iron sesquioxide 1 part Citric anhydride 0.2 parts 1997; 28: 23-32), and suspended and diluted with distilled Magnesium Stearate 1 part water. In this experiment, ramosetron was used and expressed as its hydrochloride form. All the test substances were orally administered in a dose of 12 mL/kg. In combined adminis 0057 Ten parts of maltose, 0.0008 parts of ramosetron tration of ramosetron and polycarbophil, a mixed solution of hydrochloride, 1 part of red iron sesquioxide and 0.2 parts of the two drugs was prepared and this was orally administered. citric anhydride were suspended in 67 parts of water with (2) Effect for Restraint Stress Induced Diarrhea in Rats: stirring with a magnetic stirrer to prepare a spray liquid (con 0062 Male Wistar rats (13 to 14-week age, Japan SLC centration, 15% by weight). Next, 88 parts of mannitol was Inc.) that had been fasted overnight were stressed by confine fed into a fluidized bed granulator (FLOW COATER, by ment in a restraint stress cage (KN-468, Natsume Seisakusho Freund), and the above spray liquid was sprayed onto it at a Co., Ltd.) (Hirata et al., Neurogastroenterol. Motil. 2008; 20: suction temperature of 50° C. and at a spraying speed of 10 557-565). The condition of the stool excreted within 3 hours g/min in a cycle of spraying/drying/shaking of 15 seconds/15 after the initiation of the restraint stress was observed, and the seconds/10 seconds for fluidized bed granulation. After the occurrence of diarrhea was defined in case where the excreted granulation, the granulated product was dried at a Suction stool condition was wet, amorphous and muddy stool. The effect of ramosetron or polycarbophil alone was investigated temperature of 40° C. for 5 minutes, and then mixed with 1 with the following group composition (n=12 in every group): part of magnesium Stearate. Using a rotary tabletter, the Distilled water administration group (control group); ramo mixed powder was tabletted into tablets each weight 120 mg setron administration group (0.1, 0.3 and 1 g/kg); polycar and having an initial hardness of about 1 kp. This was stored bophiladministration group (100, 300 and 1,000 mg/kg). The at 25°C. and at a relative humidity of 75% for 18 hours, and effect of combined administration of ramosetron and poly then stored at 30° C. and at a relative humidity of 40% for 4 carbophil was investigated with the following group compo hours, thereby giving oral disintegrable tablets. sition (n=24 in every group): Distilled water administration US 2011/0158930 A1 Jun. 30, 2011 group (control group); ramosetron administration group (0.3 ug/kg, q.d. (at noon) administration group; polycarbophil ug/kg); polycarbophil administration group (300 mg/kg); (300 and 1,000 mg/kg, t.i.d.)+ramosetron 100 ug/kg, q.d. (at ramosetron (0.3 ug/kg)+polycarbophil (300 mg/kg) adminis noon) administration group. tration group. All the test Substances were orally administered 0068. In every administration protocol, spontaneous def to the rat 4 hours before the initiation of the restrained stress. ecation was measured from 1 h after the animal room changed light period. Specifically, in the experiment of repeated (3) Effect for 5-HT Induced Mouse Diarrhea: administration, the lighting cycle in the animal room was so modified that the light period start from about 1 hour before 0063 Each test substance was orally administered to male the day-time administration, and under the condition, the ICR mice (9-week age, Clea Japan, Inc.), and after 2 hours, animals were habituated (1 week), and then used for admin 5-HT (3 mg/kg) was intraperitoneally administered (Miyata istration and measurement. et al., J. Pharmacol. Exp. Ther. 1992:261:297-303). The mice were put individually in separate cages, and the condition of (5) Statistical Analysis: the stool excreted within 3 hours after the 5-HT administra 0069 All the results were statistically analyzed using Sta tion was observed. The occurrence of diarrhea was defined in tistical Analysis System ver, 8.2 (SAS Institute Japan). In this case where the excreted stool condition was loose stool (wet experiment, all the data were expressed as two significant pasty stool) or watery stool (amorphous muddy stool). The figures. In the investigation using the diarrheal models, the effect of ramosetron or polycarbophil alone was investigated incidence of diarrhea was calculated in each group, then the with the following group composition (n=12 in every group): significance was evaluated according to the Fisher's exact Distilled water administration group (control group); ramo test, and if desired, the significance level was corrected setron administration group (10.30 and 100 ug/kg); polycar according to the Bonferroni’s method for consideration of bophiladministration group (100, 300 and 1,000 mg/kg). The multiplicity (the case of P-0.05 or PK0.05/3 was given sig effect of combined administration of ramosetron and poly nificance). In the experiments of spontaneous defecation, the carbophil was investigated with the following group compo mean value of total stool weight-ESEM was calculated in sition (n=24 in every group): Distilled water administration every group, and the significance was evaluated according to group (control group); ramosetron administration group (30 Student's t-test (between two groups) or Dunnett's multiple ug/kg); polycarbophil administration group (300 mg/kg); comparison test (between multiple groups) (the case of P<0. ramosetron (30 g/kg)+polycarbophil (300 mg/kg) adminis 05 was given significance). tration group. II. Results: (4) Effect on Spontaneous Defecation in Mice: 1. Effect for Restraint Stress Induced Diarrhea in Rats: 0064. In this experiment, male ddy mice (Nippon SLC, 0070. In the control group, 3-hour restraint stress caused 5-week age) were used. Spontaneous defecation was mea diarrhea at the incidence of higher than 90% in rats. Oral Sured individually in separate cages with a wire flooring administration of ramosetron (0.1 to 1 Lug/kg) and polycarbo under free drinking and free feeding condition. To avoid the phil (100 to 1,000 mg/kg) alone dose-dependently inhibited novelty stress, the mice were individually housed in the restraint stress induced diarrhea, and provided a significant respective cages with a wire flooring, from one day before the inhibitory effect with dosage of 0.3 and 1 g/kg, and 300 and test date. 1,000 mg/kg, respectively (FIG. 1A). 0065. The effect of single administration of ramosetron or 0071. The incidence of diarrhea in 0.3 ug/kg of ramosetron polycarbophil alone was investigated with the following or 300 mg/kg of polycarbophil alone was 46% and 58%, group composition, and the total stool weight in 2 hours after respectively (FIG. 1B). On the other hand, the incidence of administration of each test Substance was measured (n=10 in diarrhea in combined administration of the two was 13%; and every group): Distilled water administration group (control a significant difference was observed between the combined group); ramosetron administration group (10, 30 and 100 administration group and the respective mono-administration ug/kg); polycarbophil administration group (100, 300 and group (FIG. 1B). 1,000 mg/kg). 2. Effect for 5-HT Induced Diarrhea in Mice: 0066. The effect of combined single administration of 0072. In the control group, intraperitoneal administration ramosetron and polycarbophil was investigated with the fol of 5-HT (3 mg/kg) caused diarrhea at the incidence of higher lowing group composition, and the total stool weight in 2 than 90% in mice. Oral administration of ramosetron (10 to hours after administration of each test Substance was mea 100 ug/kg) and polycarbophil (100 to 1,000 mg/kg) alone sured (n=10 to 14 in every group): Distilled water adminis dose-dependently inhibited 5-HT induced diarrhea, and pro tration group (control group); polycarbophil administration vided a significant inhibitory effect with dosage of 30 and 100 group (300 and 1,000 mg/kg); distilled water+ramosetron ug/kg, and 300 and 1,000 mg/kg, respectively (FIG. 2A). (100 ug/kg) administration group; polycarbophil (300 and 0073. The incidence of diarrhea in 30 g/kg of ramosetron 1,000 mg/kg)+ramosetron (100 g/kg) administration group. or 300 mg/kg of polycarbophil alone was 58% and 63%, 0067. The effect of combined repeated administration of respectively (FIG. 2B). On the other hand, the incidence of ramosetron and polycarbophil was investigated by orally diarrhea in combined administration of the two was 29%; and administering to the mice the following test Substance three a significant difference was observed between the combined times a day (morning, noon, evening) for 4 days followed by administration group and the respective mono-administration measuring the total weight of the stool excreted for 2 hours group (FIG. 2B). after the drug administration at the noon on day 4 (n=10 in every group): Distilled water administration group (control 3. Effect for Spontaneous Defecation in Mice: group, t.i.d.); polycarbophil administration group (300 and 0074. In the control group, the stool weight of spontaneous 1,000 mg/kg, t.i.d.), distilled water (t.i.d.)+ramosetron 100 defecation in the mice for 2 hours after the oral administration US 2011/0158930 A1 Jun. 30, 2011
of distilled water was ranged from 140+22 to 190+23 mg daily dose or an equivalent molar amount of ramosetron or (FIGS. 3, 4 and 5). Oral administration of ramosetron (10 to other pharmaceutically acceptable salt thereof. 100 lug/kg) alone dose-dependently inhibited spontaneous 3. The method of claim 1 comprising administering to the defecation; and in the 100 lug/kg administration group, a patient 1.0 to 5.0 g of polycarbophil calcium as a daily dose or significant reduction in the total stool weight was observed as an equivalent molar amount of polycarbophil or other phar compared with that in the distilled water administration group maceutically acceptable salt thereof. (FIG. 3). On the other hand, oral administration of polycar 4. Use of ramosetron or a pharmaceutically acceptable salt bophil (100 to 1,000 mg/kg) alone had no significant influ for the manufacture of a medicament for the treatment of ence on spontaneous defecation (FIG. 3). irritable bowel syndrome with diarrhea or mixed irritable 0075. The total stool weight for 2 hours after single com bowel syndrome in combination with polycarbophil or a bined administration of 300 or 1,000 mg/kg of polycarbophil pharmaceutically acceptable salt thereof. and 100 g/kg of ramosetron was on the same level as that in single administration of 100 g/kg of ramosetron alone; and 5. Use of ramosetron hydrochloride in a daily dose of 0.001 there was not recognized any significant difference between to 0.05 mg. or an equivalent molar amount of ramosetron or the two groups (FIG. 4). other pharmaceutically acceptable salt thereof, for the manu 0076 Four-day repeated oral administration of polycarbo facture of a medicament for the treatment of irritable bowel phil, 300 and 1,000 mg/kg (t.i.d.) alone slightly increased the syndrome with diarrhea or mixed irritable bowel syndrome in stool weight in spontaneous defecation in mice; however, at combination with polycarbophil or a pharmaceutically any dose, no significant difference was observed between the acceptable salt thereof. polycarbophil group and the distilled water group (FIG. 5). 6. A pharmaceutical composition comprising ramosetron On the other hand, repeated combined administration of poly or a pharmaceutically acceptable salt thereof and polycarbo carbophil, 300 or 1,000 mg/kg (t.i.d.), and ramosetron, 100 phil or a pharmaceutically acceptable salt thereof. ug/kg (q.d.) attenuated the inhibitory effect of ramosetron on 7. The pharmaceutical composition of claim 6, comprising spontaneous defecation; and in the repeated combined a) 0.001 to 0.05 mg of ramosetron hydrochloride as a daily administration group with ramosetron and polycarbophil. dose or an equivalent molar amount of ramosetron or other 1,000 mg/kg (t.i.d.), a significant increase was observed as pharmaceutically acceptable salt thereof, and b) 1.0 to 5.0 g of compared with that in the repeated administration group with polycarbophil calcium as a daily dose or an equivalent molar ramosetron alone (FIG. 5). amount of polycarbophil or other pharmaceutically accept able salt thereof. INDUSTRIAL APPLICABILITY 8. The pharmaceutical composition of claim 7, wherein the 0077 According to the invention, there is provided a more composition is used for the treatment of irritable bowel syn excellent treatment method for patients suffering from IBS drome with diarrhea or mixed irritable bowel syndrome. with diarrhea or mixed IBS. 9. An enhancer of effect of polycarbophil or a pharmaceu 1. A method for treatment of a patient suffering from irri tically acceptable salt thereof for improving diarrhea Symp table bowel syndrome with diarrhea or mixed irritable bowel tom of irritable bowel syndrome, which comprises ramo syndrome, which comprises administering to the patient a setron or a pharmaceutically acceptable Salt thereof. therapeutically effective amount of ramosetron or a pharma 10. The enhancer of claim 9, comprising 0.001 to 0.05 mg ceutically acceptable salt thereof in combination with a thera of ramosetron hydrochloride as a daily dose or an equivalent peutically effective amount of polycarbophilor a pharmaceu molar amount of ramosetron or other pharmaceutically tically acceptable salt thereof. acceptable salt thereof. 2. The method of claim 1 comprising administering to the patient 0.001 to 0.05 mg of ramosetron hydrochloride as a c c c c c