JP2010031063A - Medicinal composition - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a new compound having action on the improvement of gastrointestinal motility function and suppressing occurrence of side effects. <P>SOLUTION: There is provided the following new compound which is rich in binding affinity to serotonin receptor 4 (5HT<SB>4</SB>) and does not cause the onset of arteritis, clot formation or the like: 4-amino-5-chloro-2-methoxy-N-[(2S, 4S)-1-ethyl-2-hydroxymethyl-4-pyrrolidinyl]benzamide, or a metabolite of its acid additive salt of 4-amino-5-chloro-2-methoxy-N-[(2S, 4S)-2-hydroxymethyl-4-pyrrolidinyl]benzamide. <P>COPYRIGHT: (C)2010,JPO&INPIT

Description

  This invention is suitable for activating the gastrointestinal tract, especially the stomach movement, and quickly eliminating abnormal residence of the ingested food in the organ, with a peripheral point of action, such as arteritis, etc. A compound having no side effect, that is, 4-amino-5-chloro-2-methoxy-N-[(2S, 4S) -2-hydroxymethyl-4-pyrrolidinyl] benzamide or an acid addition salt thereof is used as an active ingredient. Gastrointestinal tract function-improving agent for humans or animals, a pharmaceutical composition comprising this and a pharmaceutically acceptable carrier, and a gastrointestinal tract comprising administering a composition containing an effective amount of the compound to a patient The present invention relates to a method for ameliorating motor dysfunction and the use of the compound for producing the composition.

A compound whose general name is metoclopramide is widely known as a compound having a property of promoting the motor function of the stomach, but induces extrapyramidal symptoms and other unfavorable symptoms due to its action on the central nervous system. In addition, a compound having the generic name cisapride was also practically used as a gastrointestinal motility activator, but its use was stopped due to induction of ventricular arrhythmia.
4-amino-5-chloro-2-methoxy-N-[(2S, 4S) -1-ethyl-2-hydroxymethyl, which has no effect on the central nervous system or is very weak -4-Pyrrolidinyl] benzamide (sometimes described as TKS159) or an acid addition salt thereof is known as a compound having an action of enhancing the motility function of the digestive tract, particularly the stomach (Patent Literature). 1).

Japanese Patent Laid-Open No. 5-17434

  However, the applicant of the present application has developed TKS159, which is a representative compound of the invention described in the above publication, and in the case of repeated oral administration of TKS159 in a safety test using a beagle dog, thrombus formation, arteritis, We saw the findings of symptoms such as brain softening. The manifestation of such symptoms clearly shows that it is not suitable for use as a medicine.

  The present inventors have found that these various symptoms that did not appear in mice and rats are metabolites of the compound specifically produced in beagle dogs. 4-amino-5-chloro-2-methoxy-N is considered as a finding of symptoms caused by methoxy-N-[(2S, 4S) -2-hydroxymethyl-4-pyrrolidinyl] benzamide or an acid addition salt thereof. -[(2S, 4S) -2-hydroxymethyl-4-pyrrolidinyl] benzamide or its acid addition salt was used for repeated oral administration to beagle dogs. Surprisingly, when TKS159 was administered It was found that there was no evidence of the various symptoms seen. In addition, 4-amino-5-chloro-2-methoxy-N-[(2S, 4S) -2-hydroxymethyl-4-pyrrolidinyl] benzamide or an acid addition salt thereof is equivalent to TKS159 or an acid addition salt thereof. Has been found to be a compound with an ability to improve gastrointestinal motility functions. That is, 4-amino-5-chloro-2-methoxy-N-[(2S, 4S) -2-hydroxymethyl-4-pyrrolidinyl that can avoid the expression of symptoms such as thrombus formation, arteritis, brain softening, etc. It has been found that benzamide or an acid addition salt thereof can be effectively used as an agent for improving gastrointestinal motility function using this as an active ingredient. What should be desired as an attribute of a drug used for treatment is, of course, that it is sufficiently equipped with the required action, and it is unavoidable that it is inevitably associated with the drug. There is a widespread need to provide drugs that eliminate the manifestation of possible undesirable effects. In diseases other than those that directly affect life, this tendency is even stronger. There are many examples of therapeutic drugs that have not been able to fulfill their role, because they have the necessary actions, but some of the undesirable effects that cannot be avoided cannot be avoided. What is doing is well known. In the above publication, 4-amino-5-chloro-2-methoxy-N- (5-hydroxymethylpyrrolidin-3-yl) benzamide is described, but 4-amino-5-chloro-2-methoxy is described. -N-[(2S, 4S) -2-hydroxymethyl-4-pyrrolidinyl] benzamide is a new compound in stereoisomerism or optical isomerism.

  The present invention has been made on the basis of the above knowledge, and includes 4-amino-5-chloro-2-methoxy-N-[(2S, 4S) -2-hydroxymethyl-4-pyrrolidinyl] benzamide or an acid thereof. The present invention relates to an agent for improving gastrointestinal motility function which avoids side effects such as arteritis containing an addition salt as an active ingredient, and further relates to the 4-amino-5-chloro-2-methoxy-N-[(2S, 4S)- 2-Hydroxymethyl-4-pyrrolidinyl] benzamide or an acid addition salt thereof and a pharmaceutically acceptable carrier, a novel pharmaceutical for administration to a human or mammal (eg, dog, cat, cow, horse, sheep, etc.) It relates to a composition.

  The gastrointestinal motility function improving agent of the present invention or a novel pharmaceutical composition containing the same includes thrombus formation, arteritis, etc. 4-amino-5-chloro-2-methoxy-N-[(2S, 4S) -1-ethyl- 2-hydroxymethyl-4-pyrrolidinyl] benzamide or a compound capable of avoiding the side effects accompanying acid addition salts thereof, ie, 4-amino-5-chloro-2-methoxy-N-[(2S, 4S)- The present invention relates to a novel pharmaceutical composition comprising 2-hydroxymethyl-4-pyrrolidinyl] benzamide (sometimes referred to as TM161) or an acid addition salt thereof and a pharmaceutically acceptable carrier.

The compound TM161 or its acid addition salt is useful for living body, especially beagle dogs, in 4-amino-5-chloro-2-methoxy-N-[(2S, 4S) -1-ethyl-2-hydroxymethyl-4- Surprisingly, 4-amino-5-chloro-2-methoxy-N-[(2S, 4S) is found as a metabolite when a pyrrolidinyl] benzamide compound or an acid addition salt thereof is administered. ) -1-ethyl-2-hydroxymethyl-4-pyrrolidinyl] benzamide has the property of avoiding the onset of side effects such as thrombus formation and arteritis that are inevitably accompanied. Revealed in these studies. In addition, it was revealed that the ability to improve gastrointestinal motility function is superior to TKS159 or acid addition salts thereof. Furthermore, 4-amino-5-chloro-2-methoxy-N-[(2S, 4S) -2-hydroxymethyl-4-pyrrolidinyl] benzamide or an acid addition salt thereof is added to the serotonin 4 (5HT ₄ ) receptor. bonds, other receptors, for example, has also been shown to be one that acts as the preferred agonist for binding to dopamine D ₂ receptors.

Thus, there is a caused by binding to the dopamine D _2, calming effect extrapyramidal symptoms, side effects hypersecretion etc. prolactin was revealed also be a compound capable of reducing.
The present invention has been made on the basis of such various new findings. According to the present invention, 4-amino-5-chloro-2-rich having a high binding affinity for serotonin receptor 4 (5HT ₄ ). Methoxy-N-[(2S, 4S) -2-hydroxymethyl-4-pyrrolidinyl] benzamide or an acid addition salt thereof is used as an active ingredient, and 4-amino-5-chloro-2-methoxy-N-[(2S, 4S ) -1-Ethyl-2-hydroxymethyl-4-pyrrolidinyl] benzamide or its acid addition salt inevitably accompanied by gastrointestinal motility function which can avoid the occurrence of side effects such as thrombus formation and arteritis A pharmaceutical composition comprising an improving agent or an active ingredient thereof and a pharmaceutically acceptable carrier is provided. The significance of the present invention is important in view of the necessity of avoiding side effects as much as possible as an attribute of pharmaceutical compounds.

  This is because a true pharmaceutical invention is established only when it is confirmed that not only a useful pharmacological activity but also no serious side effects. That is, the present invention relates to a gastrointestinal motility function improving agent that has been confirmed to be effective and a safe drug that does not cause side effects.

The present invention
(1) 4-amino-5-chloro-2-methoxy-N-[(2S, 4S) -2, which has high binding affinity for serotonin receptor 4 (5HT ₄ ) and does not cause arteritis, thrombus formation, etc. -Hydroxymethyl-4-pyrrolidinyl] benzamide or an acid addition salt thereof as an active ingredient
(2) 4-amino-5-chloro-2-methoxy-N-[(2S, 4S) -2, which has high binding affinity for serotonin receptor 4 (5HT ₄ ) and does not cause arteritis, thrombus formation, etc. -Hydroxymethyl-4-pyrrolidinyl] benzamide or an acid addition salt thereof as an active ingredient, and a pharmaceutical composition for improving gastrointestinal motility function comprising a pharmaceutically acceptable carrier,
(3) 4-amino-5-chloro-2-methoxy-N-[(2S, 4S) -2, which has high binding affinity for serotonin receptor 4 (5HT ₄ ) and does not cause arteritis, thrombus formation, etc. -Hydroxymethyl-4-pyrrolidinyl] benzamide or an acid addition salt thereof as an active ingredient is a gastrointestinal motility improving agent or a gastrointestinal tract motility improving pharmaceutical composition comprising this and a pharmaceutically acceptable carrier. Therapeutic methods for promoting motor function,
(4) 4-amino-5-chloro-2-methoxy-N-[(2S, 4S] -2-hydroxymethyl-4-pyrrolidinyl] benzamide or an acid addition salt thereof, or a pharmaceutically acceptable carrier thereof A method for improving the motor function of a human or animal gastrointestinal tract while avoiding the onset of arteritis, thrombus formation or cerebral softening, which comprises administering a pharmaceutical composition containing the composition to a human or mammal,
(5) 4-amino-5-chloro-2-methoxy-N-[(2S, 4S] -2-hydroxymethyl-4-pyrrolidinyl) benzamide or an acid addition salt thereof,
(6) 4-amino-5-chloro-2-methoxy-N-[(2S, 4S] -2-hydroxymethyl-4 in which the amino group at the 4-position or / and the amino group of the pyrrolidinyl group may be protected -Pyrrolidinyl) benzamide or an acid addition salt thereof,
(7) 4-amino-5-chloro-2-methoxybenzoic acid or a reactive derivative thereof, wherein the amino group may be protected, and (2S, 4S) -4-amino-N-acyl-2-hydroxymethylpyrrolidine To give 4-amino-5-chloro-2-methoxy-N-[(2S, 4S) -1-acyl-2-hydroxymethyl-4-pyrrolidinyl] benzamide or an acid addition salt thereof, and then acyl 4-amino-5-chloro-2-methoxy-N-[(2S, 4S) -2-hydroxy, wherein a protecting group and an acyl group are removed when a protecting group is used as the group Methyl-4-pyrrolidinyl] benzamide or an acid addition salt thereof,
(8) The amino group may be protected (2S, 4S) -4-amino-N-acyl-2-hydroxymethylpyrrolidine,
(9) The compound according to (8), wherein the amino-protecting group is an acyl group, and the acyl group and the acyl group of N-acyl are selected from formyl, acetyl, propionyl, and benzoyl,
(10) The compound according to (8), wherein the acyl group is acetyl,
About.
Hereinafter, the present invention will be described in more detail by describing specific examples. However, the present invention is not construed as being limited or limited thereto.

FIG. 1 shows the change in binding affinity of the analyte drug to serotonin receptor 4. The horizontal axis represents the concentration of the analyte drug (molar concentration expressed in logarithm), and the vertical axis represents the binding ratio between serotonin receptor 4 and [ ³ H] GR113808. ● ── ● indicates the sample compound obtained in Example 9, and ○ ── ○ indicates TKS159 hydrochloride. As the concentration of the analyte drug increases, the amount of [ ³ H] GR113808 bound to serotonin receptor 4 decreases. That is, it is shown that the analyte drug antagonizes and binds to [ ³ H] GR113808 that binds to serotonin receptor 4. It can be seen that the affinity of the sample drug obtained in Example 9 for serotonin receptor 4 is stronger than that of TKS159 hydrochloride. FIG. 2 shows the change in binding affinity of the analyte drug to the dopamine D ₂ receptor. The horizontal axis represents the concentration of the analyte drug (molar concentration expressed in logarithm), and the vertical axis represents the binding ratio of dopamine D ₂ receptor and [ ³ H] -piperone. ● ── ● indicates the sample compound obtained in Example 9, and ○ ── ○ indicates TKS159 hydrochloride. As the concentration of the analyte drug increases, the amount of [ ³ H] -spiperone bound to the dopamine D ₂ receptor decreases. That is, it is shown that the analyte drug antagonizes and binds to [ ³ H] -sperperone that binds to the dopamine D ₂ receptor. It can be seen that the affinity of the sample drug obtained in Example 9 for the dopamine D ₂ receptor is weaker than that of TKS159 hydrochloride. FIG. 3 shows the degree of relaxation of the sample drug in the rat isolated specimen. The horizontal axis shows the concentration of the analyte drug (molar concentration expressed in logarithm), and the vertical axis shows the relaxation rate in the rat isolated specimen. ● ── ● indicates the sample compound obtained in Example 9, and ○ ── ○ indicates TKS159 hydrochloride. It can be seen that as the concentration of the analyte drug increases, relaxation of the sample occurs depending on the concentration, and the action of the drug obtained in Example 9 is stronger than that of TKS159 hydrochloride.

（式中、Ｒ^１は保護されていてもよいアミノ基を示す。）で表されるアミノ基が保護されていてもよい４−アミノ−５−クロロ−２−メトキシ安息香酸又はその反応性誘導体と、式（II） 4-Amino-5-chloro-2-methoxy-N-[(2S, 4S) -2-hydroxymethyl-4-pyrrolidinyl] benzamide or an acid addition salt thereof of the present invention has the formula (I)

(Wherein R ¹ represents an amino group which may be protected) 4-amino-5-chloro-2-methoxybenzoic acid or a reactive derivative thereof, wherein the amino group represented by And the formula (II)

（式中、Ｒ^２は水素原子又はイミノ基の保護基を示す。）で表されるイミノ基が保護されていてもよい（２Ｓ，４Ｓ）−４−アミノ−２−ヒドロキシメチルピロリジンとを反応させて式（III）

(In the formula, R ² represents a hydrogen atom or an imino group protecting group.) The imino group represented by (2S, 4S) -4-amino-2-hydroxymethylpyrrolidine may be reacted. Let formula (III)

（式中、Ｒ^１とＲ^２は前記と同意義。）で表される化合物を製造し、所望によりアミノ基の保護基又は／及びイミノ基の保護基を除去することにより製造される。

(Wherein R ¹ and R ² have the same meanings as described above) are produced, and the amino protecting group and / or the imino protecting group are optionally removed.

  As described above, the compound represented by the formula (III) in which at least one of the amino group and / or imino group is protected with a protecting group is a novel compound, 4-amino-5-chloro-2-methoxy- N-[(2S, 4S) -2-Hydroxymethyl-4-pyrrolidinyl] benzamide (TM161) or an acid addition salt thereof is not only important as an intermediate for synthesis, but also an amino protecting group or / and an imino group. When the protecting group is removed in vivo after administration of compound (III) in vivo, digestion superior to TKS159 while avoiding side effects such as thrombus formation, arteritis, cerebral softening as in TM161 Since it has an effect of improving the tube motor function, it can be used in the same manner as TM161. Examples of the protecting group for amino group and protecting group for imino group include acyl group (for example, acetyl group), BOC group, benzyloxycarbonyl group and the like. Protecting groups that can be removed in vivo by protecting amino groups and / or imino groups have been well established in the past, and in the present invention, such protecting groups that can be removed in vivo are adopted as protecting groups. Also good. Specifically, examples of protecting groups that can be removed in vivo include acyl groups (for example, lower alkylcarbonyl groups such as acetyl and propionyl), alkyloxycarbonyl groups (for example, lower alkyloxy groups such as methyloxycarbonyl and ethyloxycarbonyl). A carbonyl group) and the like are preferable.

4-amino-5-chloro-2-methoxy-N-[(2S, 4S) -2- with high binding affinity for serotonin receptor 4 (5HT ₄ ) of the present invention, which does not cause arteritis, thrombus formation, etc. Hydroxymethyl-4-pyrrolidinyl] benzamide or its acid addition salt is 4-amino-5-chloro-2-methoxy-N-[(2S, 4S) -1-ethyl-2-hydroxymethyl-4-pyrrolidinyl] benzamide. Or it is a metabolite of its acid addition salt, and this is more preferably produced as follows. That is, 4-amino-5-chloro-2-methoxybenzoic acid or a derivative thereof with a protected amino group or a reactive derivative thereof (for example, acid halide, active ester, acid anhydride, etc.) and (2S, 4S) ) -4-amino-N-acyl-2-hydroxymethylpyrrolidine is optionally subjected to a condensation reaction, optionally using a condensing agent. Examples of the acyl group include formyl, acetyl, propionyl, benzoyl and the like, and acetyl is preferred. The protecting group for the amino group may be the acyl group.

  The obtained compound may be obtained as an acid addition salt depending on the condensation method. However, when the base is derived into an acid addition salt, it is appropriately dissolved in a solvent, and the desired acid is added to the acid addition salt. Can be guided to. Here, as an appropriate medium used for the condensation reaction, tetrahydrofuran, dioxane, benzene, toluene, petroleum hydrocarbons (hexane, heptane, octane, petroleum benzine, etc.), dimethylformamide, pyridine, triethylamine, acetonitrile, Use an inert material such as chloroform or a condensing agent. In the case of using 4-amino-5-chloro-2-methoxybenzoic acid instead of a reactive derivative, the acid halide as a derivative is thionyl chloride, acid chloride or acid bromide with phosphorus tribromide, and the acid anhydride is Mixed acid anhydride with chlorocarbonic acid ethyl ester, active ester is an acid obtained by reaction with ester with ethyl alcohol or p-nitrophenol, N, N-dicarbonyldiimidazole, N, N-carbonyldipyrrole Examples include imidazolide and acid pyrrolizolide. When these reactive derivatives are used for the reaction, it is preferable to use a base such as pyridine, picoline, N-ethylmorpholine, triethylamine, or potassium carbonate. The case where a condensing agent is used in some cases refers to the case where 4-amino-5-chloro-2-methoxybenzoic acid is used for the reaction without being converted to a reactive derivative. In such a case, the condensing agent is used. . As the condensing agent used, N, N-dicyclohexylcarbodiimide, 1,1-sulfinyldiimidazole, 1,1-carbonyldiimidazole, titanium tetrachloride, phosphorus trichloride, phosphorus oxychloride, diethyl chlorophosphite, o- Examples include phenylene chlorophosphite.

  The condensation reaction can be suitably carried out with stirring at room temperature or under heating. The 4-amino-5-chloro-2-methoxy-N-[(2S, 4S) -1-acyl-2-hydroxymethyl-4-pyrrolidinyl] benzamide thus obtained may optionally be formed as an acid addition salt. However, when it is produced as a base, it is derived into an appropriate acid addition salt. Acids to be derived into such acid addition salts include hydrogen chloride, hydrogen bromide, sulfuric acid, hydrogen iodide, carbonic acid, phosphoric acid, formic acid, acetic acid, propionic acid, glycolic acid, glucuronic acid, maleic acid, glutamic acid, Examples include methanesulfonic acid, ethanesulfonic acid, toluenesulfonic acid, benzenesulfonic acid, benzoic acid, salicylic acid, phenylacetic acid, mandelic acid, lactic acid, succinic acid, tartaric acid, fumaric acid, citric acid and the like. Derivation to the acid addition salt involves the formation of 4-amino-5-chloro-2-methoxy-N-[(2S, 4S) -1-acyl-2-hydroxymethyl-4-pyrrolidinyl] benzamide as a base as described above. A desired acid addition salt can be obtained by mixing and stirring the desired acid in an appropriate solvent. Acid addition salts are usually pharmacologically acceptable salts.

  The 4-amino-5-chloro-2-methoxy-N-[(2S, 4S) -1-acyl-2-hydroxymethyl-4-pyrrolidinyl] benzamide or acid addition salt obtained here is an acyl group. 4-amino-5-chloro-2-methoxy-N-[(, which can avoid side effects such as arteritis, thrombus formation, and brain softening that are inevitably expressed in TKS159 or its acid addition salt through elimination. 2S, 4S) -2-hydroxymethyl-4-pyrrolidinyl] benzamide or an acid addition salt thereof. Here, elimination of the acyl group is achieved by adding sodium hydroxide, potassium hydroxide, etc. in alcohol (methyl alcohol, ethyl alcohol, propyl alcohol, etc.) and heating to reflux.

Thus, a gastrointestinal motility function improving agent having a high binding affinity to serotonin receptor 4 (5HT ₄ ) and capable of avoiding side effects accompanying TKS159 or acid addition salts thereof such as arteritis and thrombus formation, ie, 4 -Amino-5-chloro-2-methoxy-N-[(2S, 4S) -1-ethyl-2-hydroxymethyl-4-pyrrolidinyl] benzamide or a metabolite of its acid addition salt, 4-amino-5- Chloro-2-methoxy-N-[(2S, 4S) -2-hydroxymethyl-4-pyrrolidinyl] benzamide or an acid addition salt thereof can be obtained.

  In addition, (2S, 4S) -4-amino-2-hydroxymethylpyrrolidine in which the amino group used in the above production method is protected can be produced by the following method. A protecting group is introduced into the imino group of 4-hydroxy-L-proline alkyl ester to produce N-protected-4-hydroxy-L-proline alkyl ester, and thus produced N-protected-4-hydroxy N-protected-4-mesyloxy-L-proline alkyl ester was prepared by converting the hydroxyl group of -L-proline alkyl ester to mesyloxy group, and thus prepared N-protected-4-mesyloxy-L- Conversion of the mesyloxy group of proline alkyl ester to an azide group to produce N-protected-4-azido-L-proline alkyl ester, thus prepared N-protected-4-azido-L-proline alkyl ester Is subjected to a reduction reaction to produce N-protected- (2S, 4S) -4-azido-2-hydroxymethylpyrrolidine. The N-protected- (2S, 4S) -4-azido-2-hydroxymethylpyrrolidine thus prepared was further subjected to a reduction reaction to protect the (2S, 4S) -4-amino- 2-Hydroxymethylpyrrolidine can be produced industrially advantageously.

  As the protecting group in this production method, the above-described imino protecting group is used, and preferably an acetyl group. The alkyl group is preferably a lower alkyl group having 1 to 4 carbon atoms, and examples thereof include a methyl group and an ethyl group. The first reduction reaction is preferably performed using sodium borohydride as a reducing agent, and the second reduction reaction is preferably performed by catalytic reduction.

It is a gastrointestinal motility function improving agent that is rich in binding affinity for serotonin receptor 4 (5HT ₄ ) obtained here and can avoid side effects accompanying TKS159 or its acid addition salts such as arteritis and thrombus formation. Certain 4-amino-5-chloro-2-methoxy-N-[(2S, 4S) -2-hydroxymethyl-4-pyrrolidinyl] benzamide or an acid addition salt thereof is formulated with an appropriate pharmaceutically acceptable carrier. As a pharmaceutical composition for improving gastrointestinal motility function, it is put to practical use.

  Here, as a suitable pharmaceutically acceptable carrier, excipients such as lactose, glucose, potato starch, corn starch, carboxymethylcellulose, crystalline cellulose, light anhydrous silicic acid, etc., disintegrant starch, carboxymethylcellulose calcium and the like, Lubricants such as magnesium stearate, purified talc, calcium stearate, etc., and binders such as starch paste, hydroxypropylcellulose, carboxymethylcellulose, gum arabic, gelatin, hydroxypropylmethylcellulose, etc. Examples include flavoring agents. These are selected and formulated according to the desired dosage form.

It is a gastrointestinal motility function improving agent that is rich in binding affinity for serotonin receptor 4 (5HT ₄ ) in the present invention and can avoid side effects accompanying TKS159 or its acid addition salts such as arteritis and thrombus formation. Formulated gastrointestinal motility improvement comprising amino-5-chloro-2-methoxy-N-[(2S, 4S) -2-hydroxymethyl-4-pyrrolidinyl] benzamide or an acid addition salt thereof and an appropriate carrier Examples of the preparation that is a pharmaceutical composition include tablets, capsules, fine granules, granules, injections, syrups, and dry syrups. A carrier suitable for each of these preparations is selected from the carriers described above and used. For example, when the preparation that is formulated and is a pharmaceutical composition for improving gastrointestinal motility function is a tablet, it has a high binding affinity for serotonin receptor 4 (5HT ₄ ), such as arteritis, thrombus formation, etc. 4-amino-5-chloro-2-methoxy-N-[(2S, 4S) -2-hydroxymethyl-4-pyrrolidinyl] benzamide, or a gastrointestinal motility function improving agent capable of avoiding incidental side effects A predetermined amount of acid addition salt is put into a fluidized bed granulator together with predetermined amounts of lactose and crystalline cellulose, and granulation is performed while spraying an aqueous solution of a binder. Next, a disintegrant and a lubricant are added and mixed. The granulated product obtained here is pressure-molded by a tableting machine so as to be a tablet having a predetermined size and weight, thereby producing a tablet.

Gastrointestinal motility function that is rich in binding affinity to serotonin receptor 4 (5HT ₄ ), which is an active ingredient contained in the preparation, and can avoid side effects incidental to TKS159 or its acid addition salts such as arteritis and thrombus formation The content of 4-amino-5-chloro-2-methoxy-N-[(2S, 4S) -2-hydroxymethyl-4-pyrrolidinyl] benzamide or its acid addition salt, which is an improving agent, is determined based on the active ingredient per day. In relation to the total dose, 0.05 to 10 mg / dose. The number of administrations is adjusted by the doctor's judgment depending on the symptoms, administration period, individual sensitivity to the active ingredient, etc., but is generally administered 1 to 3 times a day.

Gastrointestinal motility function improving agent which is rich in binding affinity for serotonin receptor 4 (5HT ₄ ), which is an active ingredient in the present invention, and which can avoid side effects associated with TKS159 or its acid addition salts such as arteritis and thrombus formation 4-amino-5-chloro-2-methoxy-N-[(2S, 4S) -2-hydroxymethyl-4-pyrrolidinyl] benzamide or its acid addition salt is TKS159 in acute toxicity studies and for oral administration. Or it has less acute toxicity than its acid addition salt and is therefore suitable for oral administration. 4-amino-5-chloro-2-methoxy-N-[(2S, 4S) -1-ethyl-, which has a high binding affinity for serotonin receptor 4 (5HT ₄ ) and does not cause arteritis, thrombus formation, etc. 2-hydroxymethyl-4-pyrrolidinyl] benzamide or its metabolite of 4-amino-5-chloro-2-methoxy-N-[(2S, 4S) -2-hydroxymethyl-4-pyrrolidinyl] A gastrointestinal motility function improving agent comprising benzamide or an acid addition salt thereof as an active ingredient or a gastrointestinal motility function improving pharmaceutical composition comprising this and a carrier is provided.

[Example 1]
Measurement of the action of 4-amino-5-chloro-2-methoxy-N-[(2S, 4S) -2-hydroxymethyl-4-pyrrolidinyl] benzamide monohydrochloride on serotonin receptor 4:
The striatum extracted from the Hartley male guinea pig was homogenized in 50 mM HEPES-NaOH buffer (pH 7.4), and then centrifuged and suspended repeatedly to prepare 4 serotonin receptor preparations. The receptor preparation and [ ³ H] -GR113808 of radioligand 0.1 nM and a solution containing the sample drug obtained in Example 9 at a predetermined concentration were reacted, and a multi-filter MF-12G (glass filter ( Suction filtration was performed using Whatman GF / C), and the radioactivity of the filter paper was measured using a scintillation counter (LS6500 Beckman), and the affinity of the sample drug for serotonin receptor 4 was measured. Separately, TKS159 hydrochloride was similarly processed and the affinity was compared.
The results were as shown in FIG. IC ₅₀ was 0.25 μM, lower than 0.45 μM of TKS159 hydrochloride.

[Example 2]
4-amino-5-chloro-2-methoxy -N - [(2S, 4S) -2- hydroxymethyl-4-pyrrolidinyl] Measurement of the effect of dopamine _{D 2} receptors benzamide monohydrochloride:
The striatum extracted from the Wistar male rat was homogenized in 50 mM Tris-HCl buffer (pH 7.7), and then centrifuged and suspended repeatedly to prepare a dopamine D ₂ receptor preparation. The receptor preparation was reacted with 0.25 nM [ ³ H] -spiperone and a solution containing the sample drug obtained in Example 9 at a predetermined concentration to give a multi-filter MF-12G (glass filter ( Suction filtration was performed using Whatman GF / C), and the radioactivity of the filter paper was measured using a scintillation counter (LS6500 Beckman) to determine the affinity of the sample drug for the dopamine D ₂ receptor. Separately, TKS159 hydrochloride was similarly processed and the affinity was compared.
The results were as shown in FIG. IC ₅₀ was 34 μM, higher than 3.8 μM of TKS159 hydrochloride.

[Example 3]
Synthesis of N-acetyl-4-hydroxy-L-proline ethyl ester:
To a suspension of 600 g of 4-hydroxy-L-proline ethyl ester hydrochloride, 683 g of triethylamine and 2.4 L of chloroform, 345 g of acetic anhydride was added dropwise at 10 ° C. or lower while cooling. After stirring for 2 hours, water (0.6 L) was added for liquid separation. The organic layer was dried over magnesium sulfate and concentrated under reduced pressure to give 1020 g of N-acetyl-4-hydroxy-L-proline ethyl ester as an oil.
IR (neat) νcm ⁻¹ : 3402, 1740, 1626, 1456, 1278, 1195, 1085, 1035, 967, 864, 568

[Example 4]
Synthesis of N-acetyl-4-mesyloxy-L-proline ethyl ester:
To a solution of 1020 g of N-acetyl-4-hydroxy-L-proline ethyl ester, 435 g of triethylamine and 1.9 L of chloroform, 457 g of methanesulfonyl chloride was added dropwise at 15 ° C. or lower while cooling. After stirring for 30 minutes, 1N hydrochloric acid (0.6 L) was added to separate the layers. The organic layer was washed successively with 5% aqueous sodium hydrogen carbonate (600 g) and water (0.6 L), dried over magnesium sulfate, and concentrated under reduced pressure to give 802 g of N-acetyl-4-mesyloxy-L-proline ethyl ester. Obtained as an oil.
IR (neat) νcm ⁻¹ : 3462, 1742, 1652, 1422, 1353, 1268, 1196, 1175, 958, 905, 531

[Example 5]
Synthesis of N-acetyl-4-azido-L-proline ethyl ester:
243 g of sodium azide was added to a solution of 802 g of N-acetyl-4-mesyloxy-L-proline ethyl ester and 2.4 L of DMF, reacted at an internal temperature of 70 ° C. for 7 hours, cooled, and poured into ice water (4.8 L). Extracted with chloroform (3.2 L). The organic layer was dried over magnesium sulfate and concentrated under reduced pressure to obtain 644 g of N-acetyl-4-azido-L-proline ethyl ester as an oil.
IR (neat) νcm ⁻¹ : 3472, 2109, 1746, 1656, 1418, 1370, 1269, 1195, 1055, 1029, 615, 561

[Example 6]
Synthesis of (2S, 4S) -N-acetyl-4-azido-2-hydroxymethylpyrrolidine:
644 g of N-acetyl-4-azido-L-proline ethyl ester dissolved in 700 mL of ethanol was added dropwise to a suspension of 2.5 L of ethanol and 162 g of sodium borohydride at 10 ° C. or lower while cooling. After the reaction overnight, 35% hydrochloric acid (594 g) was added dropwise at 20 ° C. or lower while cooling, and neutralized with sodium bicarbonate (24 g). After separation by filtration, concentration under reduced pressure while replacing the solvent with 1.3 L of isopropyl alcohol gave 534 g of (2S, 4S) -N-acetyl-4-azido-2-hydroxymethylpyrrolidine as an oil.
¹ H-NMR (CDCL3) δ: 1.8 (1H, ddd), 2.1 (3H, s), 2.4 (1H, ddd), 2.9 (1H, d), 3.5 (1H , Dd), 3.8 (2H, m), 4.2 (1H, ddd), 4.3 (1H, m), 4.7 (1H, OH)
IR (neat) νcm ⁻¹ : 3371, 2104, 1626, 1445, 1362, 1327, 1269, 1048, 907, 617, 560

[Example 7]
Synthesis of (2S, 4S) -N-acetyl-4-amino-2-hydroxymethylpyrrolidine:
Add 92.4 g of 10% Pd-C to a solution of 534 g of N-acetyl-4-azido-2-hydroxymethylpyrrolidine and 2.6 L of methanol, and add hydrogen to the container every hour until the raw material disappears at normal pressure. Hydrogenation was performed with gas exchange (30 hours). After filtration, the filtrate was concentrated under reduced pressure and (2S, 4S) -N-acetyl-4-amino-2-hydroxymethylpyrrolidine, more specifically (2S, 4S)-(−)-1-acetyl-4-amino-2- 411 g of hydroxymethylpyrrolidine was obtained as an oil.
[Α] _D ²⁰ = −57.1 ° (c = 1.28, MeOH)
IR (neat) νcm ⁻¹ : 3343, 1625, 1446, 1361, 1238, 1199, 1037, 957, 915, 757, 612

[Example 8]
Synthesis of 4-acetylamino-5-chloro-2-methoxy-N-[(2S, 4S) -1-acetyl-2-hydroxymethyl-4-pyrrolidinyl] benzamide:
9.84 g of 4-acetylamino-5-chloro-2-methoxybenzoic acid and 4.5 g of triethylamine were dissolved in 40 ml of dichloromethane, and 4.60 g of ethyl chlorocarbonate was added dropwise at 10 ° C. or lower. After stirring at the same temperature for 30 minutes, a solution of 7.03 g of (2S, 4S)-(−)-1-acetyl-4-amino-2-hydroxymethylpyrrolidine in dichloromethane (20 ml) was added dropwise. After stirring at the same temperature overnight, water (20 ml) was added and the precipitated crystals were collected by filtration. The obtained crystals were dried in hot air at 50-55 ° C. and 4-acetylamino-5-chloro-2-methoxy-N. 11.74 g of [[2S, 4S) -1-acetyl-2-hydroxymethyl-4-pyrrolidinyl] benzamide was obtained.
¹ H-NMR (CDCL3) δ: 1.7 (1H, ddd), 2.1 (3H, s), 2.3 (3H, s), 2.5 (1H, ddd), 3.4 (1H , Dd), 3.7 (1H, dd), 3.9 (1H, dd), 4.0 (3H, s), 4.1 (1H, dd), 4.3 (1H, m), 4 .6 (2H, m), 7.8 (1H, s), 8.1 (1H, d), 8.2 (1H, s), 8.4 (1H, s)
¹³ C-NMR (DMSO-d6) δ: 28.13, 28.97, 38.38, 52.04, 53.24, 59.66, 61.39, 63.20, 67.02, 124.58 , 135.98, 143.55, 161.13, 161.18, 167.88, 141.17, 174.37.
IR (KBr) νcm ⁻¹ : 3251, 1697, 1629, 1563, 1511, 1457, 1397, 1309, 1238, 1194, 1080, 1049, 1013, 980, 638

[Example 9]
Synthesis of 4-amino-5-chloro-2-methoxy-N-[(2S, 4S) -2-hydroxymethyl-4-pyrrolidinyl] benzamide monohydrochloride:
11.74 g of 4-acetylamino-5-chloro-2-methoxy-N-[(2S, 4S) -1-acetyl-2-hydroxymethyl-4-pyrrolidinyl] benzamide was dissolved in ethyl alcohol (60 ml), and water was added. After adding sodium oxide (2.7 g), the mixture was heated to reflux for 8.5 hours. Water (20 ml) was added, and the mixture was stirred for 1 hour at room temperature. Insoluble matters were filtered off, and washed thoroughly with water (50 ml). The filtrate was concentrated under reduced pressure, and n-butyal alcohol (50 ml) and saturated brine (20 ml) were added to the resulting residue for liquid separation, followed by re-extraction with n-butyal alcohol (30 ml). After the extract was concentrated under reduced pressure, methyl alcohol (50 ml) was added to the residue and dissolved, and then 18% (w / w) hydrochloric acid-containing methyl alcohol (6.5 g) was gradually added to adjust the pH to 6. The crystals precipitated by cooling with ice were collected by filtration and washed with a small amount of methyl alcohol. The obtained crystals were recrystallized from ethyl alcohol, and the target compound 4-amino-5-chloro-2-methoxy-N-[(2S, 4S) -2-hydroxymethyl-4-pyrrolidinyl] benzamide monohydrochloride was obtained. 5.5 g was obtained.
mp 219-222 ° C. [α] _D ²⁰ : + 4.2 ° (c = 1.00, EtOH)
¹ H-NMR (CD ₃ OD) δ: 1.98 (1H, ddd, J = 13.6, 8.1, 5.5 Hz), 2.58 (1H, ddd, J = 13.68.3) 8.3 Hz), 3.38 (1 H, dd, J = 12.0, 4.0 Hz), 3.54 (1 H, dd, J = 12.0, 7.0 Hz), 3.79 (1 H, dd) , J = 11.5, 5.1 Hz), 3.80 (1 H, m), 3.92 (3 H, s), 3.92 (1 H, dd, J = 11.5, 3.0 Hz), 4 .68 (1H, m), 6.51 (1H, s), 7.82 (1H, s)
¹³ C-NMR (CD ₃ OD) δ: 33.17, 50.31, 52.18, 56.67, 61.03, 62.33, 98.54, 110.86, 111.62, 133.29 150.75, 159.83, 167.22.
IR (KBr) (nu) cm < ^-1 >: 3418,3385,3325,3212,2437,1637,1588,1455,1207,1161,1048,832.

[Example 10]
Using three beagle dogs as test animals, 4-amino-5-chloro-2-methoxy-N-[(2S, 4S) -2-hydroxymethyl-4-pyrrolidinyl] benzamide monohydrochloride obtained in Example 9 was used. The oral administration was repeated once a day for 4 weeks at a dose of 100 mg / kg. Thereafter, the blood was sacrificed from the carotid artery under anesthesia, and then the brain, aorta, heart, lung, liver and the like were removed, fixed with a 0.1% phosphate buffered 10% formalin solution, and stored. Each organ was embedded in paraffin and sliced to prepare a hematoxin orange-stained specimen, and then histopathological examination was performed using an optical microscope. There was no gross abnormality in any organ. Also, histopathological examination showed no abnormality, and no brain softening, arteritis, or thrombus formation was observed.

[Example 11]
Sprague-Dawly male rats, 4 weeks old, were bred for both quarantine and habituation for 8 days, and those with body weights 160.3-169.5g were grouped into 5 animals, subject group, 300 mg / kg administration group, 1000 mg It was divided into 4 groups: / kg administration group and 2000 mg / kg administration group. After grinding 4-amino-5-chloro-2-methoxy-N-[(2S, 4S) -2-hydroxymethyl-4-pyrrolidinyl] benzamide monohydrochloride obtained in Example 9 in a mortar, the compound was A sample for administration was prepared by adding 300 mg, 1000 mg, and 2000 mg in 10 ml of 0.5% aqueous methylcellulose solution (manufactured by Wako Pure Chemical Industries, Ltd., using JP Water for Injection) and stirring. Samples to be administered were prepared once a week for 7 days, and stored in a refrigerator for use. A single dose was set to 10 ml / kg, the administration route was oral administration, and gavage was performed once a day, from 9:00 to 12:00 for 28 days using a rat gastric tube. Only 0.5% methylcellulose aqueous solution was administered to the subject group at 10 ml / kg.
During the habituation and drug administration period, solid feed (CE-2 Nippon Claire) and tap water were freely ingested.
After completion of the administration period, all cases were necropsied, and macroscopic and histological observations of organs / tissues such as brain, heart, aorta, lung, pancreas, liver, vena cava were performed, and histopathological examination was performed. There was no gross abnormality in any organ. Also, histopathological examination showed no abnormality, and no brain softening, arteritis, or thrombus formation was observed.

[Example 12]
Measurement of relaxation response in rat isolated specimens of 4-amino-5-chloro-2-methoxy-N-[(2S, 4S) -2-hydroxymethyl-4-pyrrolidinyl] benzamide monohydrochloride:
About the drug obtained in Example 9, the degree of gastrointestinal motility promoting effect was measured using an esophageal specimen isolated from rats.
The esophagus in the thoracic cavity was excised from a Wistar male rat, the intrinsic muscle layer including longitudinal and circular muscles was removed, and a mucosal muscle layer specimen having a length of about 2 cm was prepared. The specimen contains nutrient solution (NaCl 118.5, KCl 4.7, CaCl ₂ 1.3, MgSO ₄ 0.6, NaHCO ₃ 25.0, KH ₂ PO ₄ 1.2, glucose 11.1 (units). After confirming the contraction of the specimen and the stability of the contraction using carbachol 3 × 10 ⁻⁶ M while flowing a 95% O ₂ /5% CO ₂ mixed gas at 32 ° C., methysergide , Ketanserin and granisetron were added at 1 μM each, and 30 minutes later, the drug obtained in Example 9 was applied cumulatively at a common ratio of 3, and the degree of relaxation was determined via a transducer to be isotonic (static tension of about 0.5 g). Separately, TKS159 hydrochloride was similarly used, and the strength of the action was compared.
The results were as shown in FIG. EC ₅₀ was 0.7 μM, lower than 1.1 μM of TKS159 hydrochloride.

[Example 13]
Using ddy male mice, the drug obtained in Example 9 and TKS159 were dissolved in 0.5% methylcellulose suspension for oral administration and physiological saline for intravenous administration. This was administered by forced oral administration using a sonde or intravenous administration, and acute toxicity was observed.

[Example 14]
50 g of the drug obtained in Example 9, 650 g of lactose and 200 g of crystalline cellulose are weighed, put into a fluidized bed granulator, and sprayed with 30 g of binder hydroxypropylcellulose as a 5% aqueous solution to obtain a granulated powder. It was. Then, 50 g of disintegrant carboxymethylcellulose calcium and 20 g of lubricant magnesium stearate were added to the granulated powder and mixed. The obtained granulated powder for tableting was pressure-molded so that the weight of one tablet was 100 mg to obtain tablets.
Separately, 48 g of hydroxypropylmethylcellulose using the tablets obtained here. A film coating solution prepared by using 7.2 g of polyethylene glycol 6000, 1.8 g of talc, 3 g of titanium oxide and 550 cc of purified water was coated until the weight of one tablet became 105 mg to obtain a film-coated tablet.

[Reference example]
TKS159 was administered orally at doses of 10, 30 and 100 mg / kg to two male Beagle dogs for 4 weeks, and observation was made. Thrombus formation in the coronary artery and slight bleeding in the perivascular space of the brain parenchyma were observed. In the surviving cases of the 100 mg / kg administration group, large thrombus formation in the left ventricle, and thrombus formation in the renal arcuate artery and interlobular artery were observed. The beagle dogs were divided into 4 groups consisting of 6, 4, 4 and 6 males and females, of which 3 groups received TKS159 at doses of 2.5, 6.0 and 15.0 mg / kg, respectively, and the control group. The vehicle was orally administered repeatedly for 13 weeks, and the general condition was observed and various tests were performed. Further, in some cases of the control group and the 15.0 mg / kg administration group, the same examination was conducted by performing a recovery test after 4 weeks of withdrawal after a repeated administration period of 13 weeks. As a result of the examination, arteritis was observed in two cases in the 15.0 mg / kg administration group, and the sites were the spinal nerve root, radial artery, cerebral puffy coat, thymus and bladder. In another example, necrosis of the arterial septum and surrounding cell infiltration were observed in the cerebellar buffy coat, lung, coronary artery, liver, bladder, stomach, vagina, intestine, diaphragm, and the like. The beagle dogs were divided into 4 groups of 5, 3, 3 and 5 for both males and females, 3 of which received TKS159 at doses of 0.25, 0.75 and 2.25 mg / kg, respectively, and the control group. The vehicle was orally administered repeatedly for 52 weeks, and the general condition was observed and various tests were performed. Further, in some cases of the control group and the 2.25 mg / kg administration group, the same examination was conducted by performing a recovery test after 4 weeks of withdrawal after a 52-week repeated administration period. As a result of the examination, in the 52-week repeated dose toxicity test, arthritis was observed in the femur bone marrow and mediastinal lymph node in 1 of 5 males in the 2.25 mg / kg group, and pear in the other 1 case. Softening was observed in the leaves and hippocampus.

In the present invention, 4-amino-5-chloro-2-methoxy-N-[(2S, 4S) -2-hydroxymethyl-4-pyrrolidinyl] benzamide having a high binding affinity for serotonin receptor 4 (5HT ₄ ). Or 4-amino-5-chloro-2-methoxy-N-[(2S, 4S) -1-ethyl-2-hydroxymethyl-4-pyrrolidinyl] benzamide or an acid addition salt thereof, using an acid addition salt thereof as an active ingredient A pharmaceutical composition comprising a gastrointestinal motility function improving agent capable of avoiding the occurrence of side effects such as thrombus formation and arteritis that are inevitably associated with the administration of the drug, or a pharmaceutically acceptable carrier comprising the same as an active ingredient Things can be provided. In view of the necessity of avoiding side effects as much as possible as an essential attribute of pharmaceutical compounds, the present invention is useful as a medicine because it is a safe drug that is effective and does not cause side effects. It is.

Claims (10)

4-amino-5-chloro-2-methoxy-N-[(2S, 4S) -2-hydroxymethyl-, which has a high binding affinity for serotonin receptor 4 (5HT ₄ ) and does not cause arteritis or thrombus formation 4-Pyrrolidinyl] benzamide or an acid addition salt thereof, an agent for improving gastrointestinal motility function. 4-amino-5-chloro-2-methoxy-N-[(2S, 4S) -2-hydroxymethyl-, which has a high binding affinity for serotonin receptor 4 (5HT ₄ ) and does not cause arteritis or thrombus formation 4-pyrrolidinyl] benzamide or an acid addition salt thereof as an active ingredient, and a pharmaceutical composition for improving gastrointestinal motility function, comprising a pharmaceutically acceptable carrier. 4-amino-5-chloro-2-methoxy-N-[(2S, 4S) -2-hydroxymethyl-, which has a high binding affinity for serotonin receptor 4 (5HT ₄ ) and does not cause arteritis or thrombus formation 4-pyrrolidinyl] benzamide or an acid addition salt thereof as an active ingredient is a gastrointestinal motility function improving agent or a gastrointestinal motility function improving pharmaceutical composition comprising this and a pharmaceutically acceptable carrier Treatment method for.   Pharmaceutical composition comprising 4-amino-5-chloro-2-methoxy-N-[(2S, 4S] -2-hydroxymethyl-4-pyrrolidinyl] benzamide or an acid addition salt thereof, and a pharmaceutically acceptable carrier A method for improving the motor function of a human or animal digestive tract while avoiding the development of arteritis, thrombus formation, or cerebral softening, characterized by administering the product to a human or mammal.   4-amino-5-chloro-2-methoxy-N-[(2S, 4S] -2-hydroxymethyl-4-pyrrolidinyl) benzamide or an acid addition salt thereof.   4-amino-5-chloro-2-methoxy-N-[(2S, 4S] -2-hydroxymethyl-4-pyrrolidinyl) in which the amino group at the 4-position and / or the amino group of the pyrrolidinyl group may be protected Benzamide or an acid addition salt thereof.   Reaction of 4-amino-5-chloro-2-methoxybenzoic acid or a reactive derivative thereof, which may have an amino group protected, with (2S, 4S) -4-amino-N-acyl-2-hydroxymethylpyrrolidine To 4-amino-5-chloro-2-methoxy-N-[(2S, 4S) -1-acyl-2-hydroxymethyl-4-pyrrolidinyl] benzamide or an acid addition salt thereof, and then protected to an acyl group 4-amino-5-chloro-2-methoxy-N-[(2S, 4S) -2-hydroxymethyl-4, characterized in that a protecting group and an acyl group are eliminated when a group is used -Pyrrolidinyl] benzamide or a method for producing an acid addition salt thereof.   (2S, 4S) -4-amino-N-acyl-2-hydroxymethylpyrrolidine in which the amino group may be protected.   The compound according to claim 8, wherein the amino-protecting group is an acyl group, and the acyl group and the N-acyl acyl group are selected from formyl, acetyl, propionyl, and benzoyl.   The compound according to claim 8, wherein the acyl group is acetyl.

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