Composition for removing helicobacter pylori comprising zasalprazan or pharmaceutically acceptable salt thereof

Abstract

The present invention relates to a composition for removing Helicobacter pylori (Helicobacter pylori), and more particularly, to a composition for removing Helicobacter pylori (Helicobacter pylori), the composition comprising zasaprazan (zasaprazan) or a pharmaceutically acceptable salt thereof.

Description

Technical Field

[0001] The present invention relates to a composition for eradicating Helicobacter pylori, which comprises zaspirone or a pharmaceutically acceptable salt thereof, and specifically, to a composition for eradicating Helicobacter pylori, which comprises zaspirone, a pharmaceutically acceptable salt thereof, a hydrate or solvate thereof, or a mixture thereof, amoxicillin, a pharmaceutically acceptable salt thereof, or a hydrate thereof, and clarithromycin or a pharmaceutically acceptable salt thereof. Background Art

[0002] Helicobacter pylori (H. pylori) is a Gram-negative spiral bacterium living in the human gastric mucosa. The World Health Organization (WHO) classifies it as a Group 1 carcinogen, and about half of the adults in Korea are infected with Helicobacter pylori. Helicobacter pylori is a bacterium that parasitizes the gastric mucosa and grows continuously. Unless appropriate treatment is received, it will not disappear on its own.

[0003] The infection route of the Helicobacter pylori bacterium is not yet clear, but it is known to spread from person to person. It is understood that the infection is closely related to the unique lifestyle of Koreans, so they are at risk of being infected with the Helicobacter pylori bacterium.

[0004] Helicobacter pylori has been identified as the causative agent of atrophic gastritis, intestinal metaplasia, peptic ulcer, mucosa-associated lymphoid tissue (MALT) lymphoma, and gastric cancer, and is also known to be related to functional dyspepsia, iron deficiency anemia of unknown cause, and chronic idiopathic thrombocytopenia, etc.

[0005] The results of a multicenter study on asymptomatic adults in Korea showed that the seroprevalence in 2015 was 51.0%, showing a steady decline compared to the past. This may be attributed not only to active treatment but also to the improvement of economic conditions and sanitation.

[0006] The first-line eradication of Helicobacter pylori is to take a gastric acid secretion inhibitor and two antibiotics (amoxicillin and clarithromycin) for 14 days. The treatment success rate is 70% to 80%, but some patients stop taking the medicine on their own, often resulting in treatment failure. In these patients, treatment difficulties may occur due to strain resistance.

[0007] Among the pathogenic factors of Helicobacter pylori, urease neutralizes gastric acid, enabling Helicobacter pylori to colonize and proliferate in a strongly acidic environment. In addition, ammonia produced by enzymatic action affects gastric epithelial cells, and various cytokines produced when Helicobacter pylori stimulates gastric epithelial cells are used to recruit various inflammatory cells.

[0008] Therefore, there is a need to develop a novel therapeutic agent that can directly exert antibacterial or urease inhibitory effects on Helicobacter pylori, as well as a triple drug regimen using the novel therapeutic agent in combination with the antibiotics amoxicillin and clarithromycin.

[0009] [Prior Art Documents]

[0010] [Patent Documents]

[0011] (Patent Document 1) Korean Patent Publication No. 10-2000-0005291 Summary of the Invention

[0012] Technical Problem

[0013] The present inventors have found that there is a need to develop a composition for eradicating Helicobacter pylori (H. pylori) by combining a novel therapeutic agent that can directly exert antibacterial or urease inhibitory effects on H. pylori with amoxicillin and / or clarithromycin. Therefore, the present inventors used zafaptrazen, its pharmaceutically acceptable salts, its hydrates or solvates, or mixtures thereof, which can exert direct antibacterial or urease inhibitory effects on H. pylori, as the novel therapeutic agent, and by using zafaptrazen in combination with amoxicillin and / or clarithromycin, it was confirmed that the activity against H. pylori was significantly enhanced due to the synergistic and complementary effects generated by their combined use. Therefore, the object is to provide a composition for eradicating bacteria by combining two or three drugs.

[0014] Technical Solution

[0015] To achieve the above object, the present invention discloses the following means.

[0016] On the one hand, the present invention discloses a composition for eradicating Helicobacter pylori (H. pylori), which comprises zafaptrazen, its pharmaceutically acceptable salts, its hydrates or solvates, or mixtures thereof, and amoxicillin, its pharmaceutically acceptable salts, or its hydrates.

[0017] On the other hand, the present invention discloses a composition for eradicating Helicobacter pylori, which comprises zafaptrazen, its pharmaceutically acceptable salts, its hydrates or solvates, or mixtures thereof, and clarithromycin or its pharmaceutically acceptable salts.

[0018] Finally, the present invention discloses a composition for eradicating Helicobacter pylori, which comprises zafaptrazen, its pharmaceutically acceptable salts, its hydrates or solvates, or mixtures thereof, amoxicillin, its pharmaceutically acceptable salts or its hydrates, and clarithromycin or its pharmaceutically acceptable salts.

[0019] Advantageous Effects

[0020] Due to the Helicobacter pylori eradication effect of zaspirone, its pharmaceutically acceptable salts, its hydrates or solvates, or mixtures thereof, the composition for eradicating bacteria of the present invention exhibits a therapeutic effect on gastroesophageal reflux disease and / or peptic ulcer, independent of Helicobacter pylori (H. pylori) infection. In particular, it shows excellent effects in patients infected with Helicobacter pylori, and in the evaluation of all symptoms (heartburn, acid reflux, and heartburn / acid reflux) caused by Helicobacter pylori infection, it tends to improve all symptoms within 24 hours and continuously for 7 days.

[0021] In addition, due to the synergistic effect of co-administration with amoxicillin, its pharmaceutically acceptable salts or its hydrates, or with clarithromycin or its pharmaceutically acceptable salts, zaspirone, its pharmaceutically acceptable salts, its hydrates or solvates, or mixtures thereof show excellent efficacy in eradicating Helicobacter pylori.

[0022] In addition, in the pharmacokinetic (PK) clinical trial of the co-administration of zaspirone, its pharmaceutically acceptable salts, its hydrates or solvates, or mixtures thereof with amoxicillin, its pharmaceutically acceptable salts, or its hydrates and clarithromycin or its pharmaceutically acceptable salts, compared with the single administration of zaspirone, its pharmaceutically acceptable salts, its hydrates or solvates, or mixtures thereof, it shows an effect of increasing the area under the concentration-time curve (AUC T ) by more than two-fold.

[0023] In other words, considering the antibacterial effect of zaspirone, its pharmaceutically acceptable salts, its hydrates or solvates, or mixtures thereof itself, the synergistic effect of the co-administration of zaspirone, its pharmaceutically acceptable salts, its hydrates or solvates, or mixtures thereof with amoxicillin, its pharmaceutically acceptable salts, or its hydrates, or with clarithromycin or its pharmaceutically acceptable salts; and the pharmacokinetic (PK) results of the co-administration of zaspirone, its pharmaceutically acceptable salts, its hydrates or solvates, or mixtures thereof with amoxicillin, its pharmaceutically acceptable salts, or its hydrates, or with clarithromycin or its pharmaceutically acceptable salts, the triple-drug regimen of zaspirone, its pharmaceutically acceptable salts, its hydrates or solvates, or mixtures thereof with amoxicillin, its pharmaceutically acceptable salts, or its hydrates and clarithromycin or its pharmaceutically acceptable salts has a synergistic effect on the eradication of Helicobacter pylori.

[0024] The effects of the present invention are not limited to the above effects, but may include various effects that are obvious to those skilled in the art from the following description. Detailed Description

[0025] Next, the present invention will be described in more detail.

[0026] This is specifically explained as follows. When considering the functions in the present invention, the terms used herein are selected from the generally most widely used general terms, but they can vary according to the intentions of those skilled in the art, precedents, the emergence of new technologies, etc. In addition, in some cases, there are terms arbitrarily selected by the applicant, and in such cases, their meanings will be described in detail in the corresponding parts of the detailed description of the present invention. Therefore, the terms used herein should be defined based on the meanings of the terms and the overall content of the present invention, rather than simply based on the names of the terms.

[0027] Unless otherwise defined, all terms used herein, including technical and scientific terms, have the same meaning as commonly understood by those of ordinary skill in the art to which this invention belongs. Terms such as those defined in commonly used dictionaries should be interpreted as having a meaning consistent with their meaning in the context of the relevant art, and should not be interpreted in an idealized or overly formal sense unless explicitly defined in the present invention.

[0028] Numeric ranges include the values defined therein. Each maximum numeric limitation given throughout this specification includes every lower numeric limitation, as if such lower numeric limitations were expressly written herein. Each minimum numeric limitation given throughout this specification includes every higher numeric limitation, as if such higher numeric limitations were expressly written herein. Each numeric limitation given throughout this specification will include every better numeric range within the broader numeric range, as if the narrower numeric limitations were expressly written herein.

[0029] Hereinafter, the various descriptions and embodiments disclosed in the present invention can also be applied to other descriptions and embodiments respectively. In other words, all combinations of the various elements disclosed in the present invention fall within the scope of the present invention. In addition, the scope of the present invention should not be considered limited by the specific descriptions given below.

[0030] As used herein, unless otherwise specifically stated in the phrase or statement including such expression, an expression such as "comprising" should be understood as an open-ended term, meaning the possibility of including other embodiments.

[0031] The term "zastaprazan" as used herein can refer to zastaprazan, its pharmaceutically acceptable salts, its hydrates or solvates, or mixtures thereof. Thus, "a composition comprising zastaprazan" as used herein can refer to a composition comprising zastaprazan, its pharmaceutically acceptable salts, its hydrates or solvates, or mixtures thereof.

[0032] In addition, the term "amoxicillin" as used herein can refer to amoxicillin, its pharmaceutically acceptable salts, or its hydrates. Thus, a "composition comprising amoxicillin" as used herein can refer to a composition comprising amoxicillin, its pharmaceutically acceptable salts, or its hydrates.

[0033] The term "clarithromycin" as used herein can refer to clarithromycin or its pharmaceutically acceptable salts. Thus, a "composition comprising clarithromycin" as used herein can refer to a composition comprising clarithromycin or its pharmaceutically acceptable salts.

[0034] Hereinafter, the present invention will be described in detail.

[0035] A composition for eradicating Helicobacter pylori, comprising zaspirone

[0036] The present invention discloses a composition for eradicating Helicobacter pylori, which comprises zaspirone and amoxicillin.

[0037] Specifically, the present invention discloses a composition for eradicating Helicobacter pylori, which comprises zaspirone, its pharmaceutically acceptable salts, its hydrates or solvates, or mixtures thereof, and amoxicillin, its pharmaceutically acceptable salts, or its hydrates.

[0038] The present invention discloses a composition for eradicating Helicobacter pylori, which comprises zaspirone and clarithromycin.

[0039] Specifically, the present invention discloses a composition for eradicating Helicobacter pylori, which comprises zaspirone, its pharmaceutically acceptable salts, its hydrates or solvates, or mixtures thereof, and clarithromycin or its pharmaceutically acceptable salts.

[0040] The present invention discloses a composition for eradicating Helicobacter pylori, which comprises zaspirone, amoxicillin and clarithromycin.

[0041] Specifically, the present invention discloses a composition for eradicating Helicobacter pylori, which comprises zaspirone, its pharmaceutically acceptable salts, its hydrates or solvates, or mixtures thereof, amoxicillin, its pharmaceutically acceptable salts, or its hydrates, and clarithromycin or its pharmaceutically acceptable salts.

[0042] The composition for eradicating bacteria of the present invention, in addition to the antibiotic amoxicillin, its pharmaceutically acceptable salts, or its hydrates, and / or clarithromycin, or its pharmaceutically acceptable salts, further comprises zaspirone, its pharmaceutically acceptable salts, its hydrates or solvates, or mixtures thereof, and exhibits a therapeutic effect on gastroesophageal reflux disease and / or peptic ulcer due to the Helicobacter pylori eradication effect of zaspirone itself. In particular, it exhibits excellent effects on patients infected with Helicobacter pylori, and in the evaluation of all symptoms (heartburn, acid reflux, and heartburn / reflux) caused by Helicobacter pylori infection, it tends to improve all symptoms within 24 hours and continuously for 7 days.

[0043] In addition, the composition for eradicating bacteria in another aspect of the present invention is for the combined use of zaspirone, its pharmaceutically acceptable salts, its hydrates or solvates, or mixtures thereof with the antibiotic amoxicillin, its pharmaceutically acceptable salts, or its hydrates, or clarithromycin or its pharmaceutically acceptable salts (dual-drug regimen), and by the combined use thereof, due to the synergistic and complementary effects produced by the combination, the activity of eradicating Helicobacter pylori is significantly enhanced.

[0044] In addition, the composition for eradicating bacteria in yet another aspect of the present invention is for the combined use of zaspirone, its pharmaceutically acceptable salts, its hydrates or solvates, or mixtures thereof with the antibiotic amoxicillin, its pharmaceutically acceptable salts, or its hydrates, and clarithromycin or its pharmaceutically acceptable salts (triple-drug regimen), and by the combined use thereof, compared with the single administration of zaspirone, its pharmaceutically acceptable salts, its hydrates or solvates, or mixtures thereof, the area under the concentration-time curve (AUC T ) increases by more than two-fold.

[0045] Therefore, considering the antibacterial (anti-bacterial) effect of zaspirone, its pharmaceutically acceptable salts, its hydrates or solvates, or mixtures thereof itself, the synergistic effect of the combined administration of zaspirone, its pharmaceutically acceptable salts, its hydrates or solvates, or mixtures thereof with amoxicillin, its pharmaceutically acceptable salts, or its hydrates, or with clarithromycin or its pharmaceutically acceptable salts, and the pharmacokinetic (PK) results of the combined administration of zaspirone, its pharmaceutically acceptable salts, its hydrates or solvates, or mixtures thereof with amoxicillin, its pharmaceutically acceptable salts, or its hydrates, or with clarithromycin or its pharmaceutically acceptable salts, it is obvious that the triple-drug regimen of zaspirone, its pharmaceutically acceptable salts, its hydrates or solvates, or mixtures thereof with amoxicillin, its pharmaceutically acceptable salts, or its hydrates and clarithromycin or its pharmaceutically acceptable salts has a synergistic effect on the eradication of Helicobacter pylori.

[0046] In the present invention, zaspirone is the active ingredient for eradicating Helicobacter pylori.

[0047] As used herein, the term "active ingredient" refers to one or a group of substances that are expected to directly or indirectly exhibit the efficacy and effect of a pharmaceutical composition through inherent pharmacological actions, and it includes the main ingredient or the active ingredient.

[0048] In the present invention, zaspirone is an example of an imidazo[1,2-a]pyridine derivative, and its chemical name is azetidin-1-yl-[8-[(2,6-dimethylbenzyl)amino]-2,3-dimethylimidazo[1,2-a]pyridin-6-yl]methanone.

[0049] The zaspirone of the present invention can exist in the form of a pharmaceutically acceptable salt, and as a salt, the acid addition salts formed from pharmaceutically acceptable free acids are useful. As used herein, the term "pharmaceutically acceptable salt" refers to any organic or inorganic acid addition salt of zaspirone, the concentration of which has a relatively non-toxic and harmless effect on the patient, and the side effects caused by the salt do not reduce the beneficial efficacy of zaspirone. Organic acids and inorganic acids can be used as free acids. Inorganic acids such as hydrochloric acid, phosphoric acid, sulfuric acid, nitric acid, or tartaric acid can be used, and organic acids such as methanesulfonic acid, p-toluenesulfonic acid, acetic acid, trifluoroacetic acid, maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, mandelic acid, propionic acid, citric acid, lactic acid, glycolic acid, gluconic acid, galacturonic acid, glutamic acid, glutaric acid, glucuronic acid, aspartic acid, ascorbic acid, carbonic acid, vanillic acid, or hydroiodic acid can be used.

[0050] The pharmaceutically acceptable salts of the present invention include salts of acidic or basic groups, and unless otherwise specified, the acidic or basic groups may be present in zaspirone. For example, pharmaceutically acceptable salts may include sodium salts, calcium salts, and potassium salts of hydroxyl groups, and other pharmaceutically acceptable salts of amino groups include hydrobromides, sulfates, bisulfates, phosphates, hydrogen phosphates, dihydrogen phosphates, acetates, succinates, citrates, tartrates, lactates, mandelates, methanesulfonates (mesylates), and p-toluenesulfonates (tosylates), etc., and can be prepared by methods known in the art for preparing salts.

[0051] Specifically, the pharmaceutically acceptable salt of zaspirone can be zaspirone citrate (azetidin-1-yl {8-[(2,6-dimethylbenzyl)amino]-2,3-dimethylimidazo[1,2-a]pyridin-6-yl}methanone citrate), but is not limited thereto.

[0052] As used herein, the term "hydrate" refers to a hydrate formed by binding the active ingredient zaspirone or a pharmaceutically acceptable salt thereof to water through non-covalent intermolecular forces, and which contains a stoichiometric or non-stoichiometric amount of water. Specifically, based on 1 mol of the active ingredient, the hydrate may contain from about 0.25 mol to about 10 mol of water, and more specifically, may contain about 0.5 mol, about 1 mol, about 1.5 mol, about 2 mol, about 3 mol, about 5 mol, etc.

[0053] As used herein, the term "solvate" refers to a compound formed by binding the active ingredient zaspirone or a pharmaceutically acceptable salt thereof to a solvent through non-covalent intermolecular forces, and which contains a stoichiometric or non-stoichiometric amount of the solvent. Preferred solvents are volatile and non-toxic and can be administered to humans in very small amounts. Examples thereof include methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol, 1-acetate, acetone, acetic acid, anisole, tetrahydrofuran, methyl acetate, ethyl acetate, propyl acetate, isopropyl acetate, isobutyl acetate, n-butyl acetate, dimethyl sulfoxide, pentane, and heptane, but the solvates of the present invention are not limited to these examples, and specifically, based on 1 mol of the active ingredient, the solvate may contain from about 0.25 mol to about 10 mol of the solvent, and more specifically, may contain about 0.5 mol, about 1 mol, about 1.5 mol, about 2 mol, about 3 mol, about 5 mol, etc.

[0054] As used herein, the term "bacterial eradication" has a meaning similar to sterilization and means the elimination of bacteria. The term means a state in which metabolism stops without directly killing the microorganisms until the microorganisms die after a certain period of time, that is, a state in which the proliferation and growth of bacteria stop. The eradication of Helicobacter pylori herein can be considered to include the elimination or eradication of Helicobacter pylori present in the human stomach, or the cessation of the proliferation and growth of Helicobacter pylori.

[0055] In the present invention, the composition may be administered one to three times a day, preferably one or two times. Specifically, zaspirone, a pharmaceutically acceptable salt thereof, a hydrate or solvate thereof, or a mixture thereof, and amoxicillin, a pharmaceutically acceptable salt thereof, or a hydrate thereof, and / or clarithromycin or a pharmaceutically acceptable salt thereof may be administered one to three times a day, preferably one or two times, for 5 to 14 days, but the administration is not limited thereto.

[0056] In the present invention, zaspirone, its pharmaceutically acceptable salts, hydrates or solvates, or mixtures thereof, calculated as zaspirone (in free base form), may contain 1 mg to 100 mg, 2 mg to 60 mg, or 3 mg to 40 mg. Specifically, zaspirone, its pharmaceutically acceptable salts, hydrates or solvates, or mixtures thereof, calculated as zaspirone (in free base form), may be contained in an amount of about 3.3 mg, 6.6 mg, 13.1 mg, 26.2 mg or 52.4 mg, and may be contained in an amount of 5 mg to 40 mg calculated as zaspirone citrate. Specifically, it may be contained in an amount of 5 mg, 10 mg, 15 mg, 20 mg or 40 mg calculated as zaspirone citrate, and more specifically, it may be contained in an amount of 10 mg, 20 mg or 40 mg calculated as zaspirone citrate. Therefore, the composition of the present invention shows a Helicobacter pylori eradication effect greater than or equal to that of Nexium tablets 40 mg, even if the composition contains a low dose (10 mg or 20 mg) of zaspirone, thereby showing a high cure (healing) rate within 24 hours after administration and for 7 days and improving symptoms such as heartburn, acid reflux and heartburn / acid reflux. In addition, even when a low dose of zaspirone is contained, due to the synergistic and complementary effects of combined use with amoxicillin, its pharmaceutically acceptable salts, or hydrates and / or clarithromycin or its pharmaceutically acceptable salts, the activity of eradicating Helicobacter pylori is significantly enhanced.

[0057] In the present invention, the content of amoxicillin, its pharmaceutically acceptable salts, or hydrates may be 100 mg to 1000 mg, specifically 375 mg to 625 mg, more specifically 500 mg, but is not limited thereto.

[0058] In the present invention, the content of clarithromycin or its pharmaceutically acceptable salts may be 100 mg to 1000 mg, specifically 125 mg to 875 mg, more specifically 500 mg, but is not limited thereto.

[0059] In the present invention, for pharmaceutically acceptable salts and hydrates of amoxicillin and clarithromycin, the same content as that described above for zaspirone (content) may be used.

[0060] In the present invention, the composition for eradicating bacteria may contain one or more pharmaceutical additives, which are composed of excipients, disintegrants, binders and lubricants, but are not limited thereto.

[0061] In the present invention, the excipient may be one or more of microcrystalline cellulose, lactose hydrate, anhydrous lactose, sucrose, D-mannitol, starch, corn starch or light anhydrous silicic acid, but is not limited thereto.

[0062] In the present invention, the disintegrant may include starch or modified starch, such as sodium carboxymethyl starch, corn starch, potato starch or pregelatinized starch; clay, such as bentonite, montmorillonite or magnesium aluminum silicate (veegum); cellulose, such as hydroxypropyl cellulose or carboxymethyl cellulose; alginates, such as sodium alginate or alginic acid; cross-linked cellulose, such as croscarmellose sodium; gums, such as guar gum or xanthan gum; cross-linked polymers, such as cross-linked povidone; effervescent agents, such as sodium bicarbonate or citric acid, etc. These may be used alone or in combination of two or more, but are not limited thereto.

[0063] In the present invention, the binder may be one or more selected from the group consisting of hydroxypropyl cellulose, hydroxypropyl methyl cellulose, polyvinyl pyrrolidone, copovidone, starch, microcrystalline cellulose, colloidal silica, mannitol, lactose, polyethylene glycol, and mixtures thereof, but is not limited thereto.

[0064] In the present invention, examples of the lubricant may include calcium stearate, glyceryl monostearate, glyceryl palmitostearate, magnesium stearate, sodium lauryl sulfate, sodium stearyl fumarate, zinc stearate, stearic acid, hydrogenated vegetable oil, polyethylene glycol, sodium benzoate and talc. These may be used alone or in combination of two or more, but are not limited thereto.

[0065] In the present invention, the composition for eradicating bacteria may be a solid oral formulation, but is not limited thereto.

[0066] In the present invention, the solid oral formulation may be one of tablets, film-coated tablets, capsules, powders, granules, pills, lozenges, oral gels and oral dissolving film preparations, but is not limited thereto.

[0067] Specifically, in the case of film-coated tablets, coating agents widely known in the art may be used, but they are not limited thereto.

[0068] A pharmaceutical composition for treating Helicobacter pylori infection, comprising zaspirone

[0069] The present invention discloses a composition for treating Helicobacter pylori infection, which comprises zaspirone, its pharmaceutically acceptable salts, its hydrates or solvates, or mixtures thereof, and amoxicillin, its pharmaceutically acceptable salts, or its hydrates.

[0070] The present invention discloses a composition for treating Helicobacter pylori infection, which comprises zaspirone, its pharmaceutically acceptable salt, its hydrate or solvate, or a mixture thereof, and clarithromycin or its pharmaceutically acceptable salt.

[0071] The present invention discloses a composition for treating Helicobacter pylori infection, which comprises zaspirone, its pharmaceutically acceptable salt, its hydrate or solvate, or a mixture thereof, amoxicillin, its pharmaceutically acceptable salt, or its hydrate, and clarithromycin or its pharmaceutically acceptable salt.

[0072] The zaspirone is the same as described above.

[0073] As used herein, the term "treatment" refers to partially or completely alleviating, improving, relieving, inhibiting or delaying the onset of Helicobacter pylori, reducing the severity, or reducing the occurrence of one or more symptoms or characteristics by administering a composition according to the present invention.

[0074] Regarding the above content of the composition for eradicating bacteria, all can be applied to the composition for treating infection as long as they are not mutually contradictory.

[0075] A method for eradicating Helicobacter pylori, comprising the step of administering a pharmaceutical composition to a subject in need thereof

[0076] The present invention provides a method for eradicating Helicobacter pylori, which comprises the step of administering a pharmaceutically effective amount of a composition to a subject in need thereof, the composition comprising a therapeutically effective amount of zaspirone, its pharmaceutically acceptable salt, its hydrate or solvate, or a mixture thereof, and amoxicillin, its pharmaceutically acceptable salt, or its hydrate.

[0077] The present invention provides a method for eradicating Helicobacter pylori, which comprises the step of administering a pharmaceutically effective amount of a composition to a subject in need thereof, the composition comprising a therapeutically effective amount of zaspirone, its pharmaceutically acceptable salt, its hydrate or solvate, or a mixture thereof, and clarithromycin or its pharmaceutically acceptable salt.

[0078] The present invention provides a method for eradicating Helicobacter pylori, which comprises the step of administering a pharmaceutically effective amount of a composition to a subject in need thereof, the composition comprising a therapeutically effective amount of zaspirone, its pharmaceutically acceptable salt, its hydrate or solvate, or a mixture thereof, amoxicillin, its pharmaceutically acceptable salt, or its hydrate, and clarithromycin or its pharmaceutically acceptable salt.

[0079] As used herein, the term "pharmaceutically effective amount" refers to an amount effective to eradicate Helicobacter pylori. For example, it refers to the amount of the composition administered to a subject, which may include all amounts of the composition that can prevent the occurrence or recurrence of Helicobacter pylori, relieve symptoms, inhibit direct or indirect pathological consequences, prevent metastasis, reduce the progression rate, alleviate or temporarily relieve the disease condition, or improve the prognosis. In other words, a pharmaceutically effective amount can be interpreted to cover all doses that can eradicate Helicobacter pylori through the composition.

[0080] Specifically, in the method for eradicating Helicobacter pylori according to the present invention, zaspirone, its pharmaceutically acceptable salts, its hydrates or solvates, or mixtures thereof, can be administered to the subject in an amount of 1 mg to 100 mg, 2 mg to 60 mg, or 3 mg to 40 mg, calculated as zaspirone (in free base form). Specifically, zaspirone, its pharmaceutically acceptable salts, its hydrates or solvates, or mixtures thereof, can be administered in an amount of about 3.3, 6.6, 13.1, 26.2, or 52.4 mg, calculated as zaspirone (in free base form), and in an amount of 5 to 40 mg, calculated as zaspirone citrate. Preferably, it is administered in an amount of 5 mg, 10 mg, 15 mg, 20 mg, or 40 mg, calculated as zaspirone citrate. More preferably, it is administered in an amount of 10 mg, 20 mg, or 40 mg, calculated as zaspirone citrate, once to three times a day, to effectively treat the subject by eradicating Helicobacter pylori.

[0081] As used herein, the term "subject" refers to a mammal, and specifically, mammals including humans include mammals such as humans, monkeys, cows, horses, dogs, cats, rabbits, rats, and mice, and more specifically, it can mean a human. The subject can be a human who is infected or suspected of being infected with Helicobacter pylori due to reasons such as gastroesophageal reflux disease, chronic gastritis, gastric or duodenal ulcers, and gastric cancer.

[0082] Here, the term "human infected with Helicobacter pylori" refers to a human who is confirmed to be infected with Helicobacter pylori through a breath or fecal test or upper endoscopy due to symptoms such as gastroesophageal reflux symptoms (heartburn, acid reflux, heartburn / acid reflux, etc.), upper abdominal pain, indigestion, or abdominal discomfort (flatulence, bloating, or burning sensation). The term "human suspected of being infected with Helicobacter pylori" refers to a situation where the above symptoms are experienced.

[0083] Regarding the above content of the composition for eradicating bacteria, it can be applied to all methods for eradicating bacteria as long as they are not contradictory.

[0084] Use for eradicating Helicobacter pylori

[0085] The present invention provides the use of a pharmaceutical composition comprising zafaptrazan, a pharmaceutically acceptable salt thereof, a hydrate or solvate thereof, or a mixture thereof, and amoxicillin, a pharmaceutically acceptable salt thereof, or a hydrate thereof, for eradicating Helicobacter pylori.

[0086] The present invention provides the use of a pharmaceutical composition comprising zafaptrazan, a pharmaceutically acceptable salt thereof, a hydrate or solvate thereof, or a mixture thereof, and clarithromycin or a pharmaceutically acceptable salt thereof, for eradicating Helicobacter pylori.

[0087] The present invention provides the use of a pharmaceutical composition comprising zafaptrazan, a pharmaceutically acceptable salt thereof, a hydrate or solvate thereof, or a mixture thereof, amoxicillin, a pharmaceutically acceptable salt thereof, or a hydrate thereof, and clarithromycin or a pharmaceutically acceptable salt thereof, for eradicating Helicobacter pylori.

[0088] Regarding the above content of the composition for eradicating bacteria, it can all be applied to the use for eradicating Helicobacter pylori as long as they are not contradictory to each other.

[0089] Use for preparing a medicament for eradicating Helicobacter pylori

[0090] The present invention provides the use of a composition comprising zafaptrazan, a pharmaceutically acceptable salt thereof, a hydrate or solvate thereof, or a mixture thereof, and amoxicillin, a pharmaceutically acceptable salt thereof, or a hydrate thereof, for preparing a drug for eradicating Helicobacter pylori.

[0091] The present invention provides the use of a composition comprising zafaptrazan, a pharmaceutically acceptable salt thereof, a hydrate or solvate thereof, or a mixture thereof, and clarithromycin or a pharmaceutically acceptable salt thereof, for preparing a drug for eradicating Helicobacter pylori.

[0092] The present invention provides the use of a composition comprising zafaptrazan, a pharmaceutically acceptable salt thereof, a hydrate or solvate thereof, or a mixture thereof, amoxicillin, a pharmaceutically acceptable salt thereof, or a hydrate thereof, and clarithromycin or a pharmaceutically acceptable salt thereof, for preparing a drug for eradicating Helicobacter pylori.

[0093] The composition for preparing a drug according to the present invention can be mixed with an acceptable carrier and the like, and can further contain other reagents.

[0094] Regarding the above content of the composition for eradicating bacteria, it can be applied to all uses for preparing a drug for eradicating Helicobacter pylori as long as they are not contradictory.

[0095] Hereinafter, the present invention will be described in more detail by using preparation examples and examples. It is obvious to those skilled in the art that these preparation examples and examples are only intended to explain the present invention more specifically, and the scope of the present invention is not limited by them.

[0096] The active ingredient used in the following preparation examples and examples is zaspirone citrate, which is referred to as zaspirone or code name JP-1366 for convenience.

[0097] Preparation Examples

[0098] Preparation Example 1. Preparation of zaspirone citrate

[0099] Azetidin-1-yl {8-[(2,6-dimethylbenzyl)amino]-2,3-dimethylimidazo[1,2-a]pyridin-6-yl} methanone citrate was obtained according to the following method. Specifically, azetidin-1-yl {8-[(2,6-dimethylbenzyl)amino]-2,3-dimethylimidazo[1,2-a]pyridin-6-yl} methanone was obtained according to the method described in Korean Patent Publication No. 10-1777971. The NMR analysis results of the obtained azetidin-1-yl {8-[(2,6-dimethylbenzyl)amino]-2,3-dimethylimidazo[1,2-a]pyridin-6-yl} methanone are as follows.

[0100] 1 H NMR(400MHz,CDCl3); δ7.63(d,J = 1.2Hz,1H),7.13(dd,J = 8.4,6.8Hz,1H),7.06 - 7.04(m,2H),6.42(d,J = 1.2Hz,1H),4.86 - 4.84(m,1H),4.41 - 4.28(m,4H),4.37(d,J = 4.4Hz,2H),3.75 - 3.69(m,1H),2.43 - 2.34(m,13H).

[0101] Next, the obtained azetidin-1-yl {8-[(2,6-dimethylbenzyl)amino]-2,3-dimethylimidazo[1,2-a]pyridin-6-yl}methanone was mixed with an alcohol solvent (isopropyl alcohol, IPA), and the resulting mixture was stirred and then dried under vacuum at about 30 to 35 °C to obtain a dry product. Approximately 10 g of the dry product was taken and stirred with approximately 167 g of acetone. Thereto, a solution prepared by dissolving citric acid (approximately 5 g) in acetone (approximately 33 g) was slowly added dropwise over 60 minutes, and the resulting mixture was stirred at the same temperature for one hour. The mixture was cooled to about 20 °C to 25 °C and stirred for another hour. The resulting solid was filtered, washed with acetone, and dried under vacuum to obtain azetidin-1-yl {8-[(2,6-dimethylbenzyl)amino]-2,3-dimethylimidazo[1,2-a]pyridin-6-yl}methanone citrate. The NMR analysis results of the above-obtained azetidin-1-yl {8-[(2,6-dimethylbenzyl)amino]-2,3-dimethylimidazo[1,2-a]pyridin-6-yl}methanone citrate (zastaprazan citrate) are described below.

[0102] 1 H NMR (400 MHz, MeOD); δ 7.90 (s, 1H), 7.06 - 7.15 (m, 3H), 6.77 (s, 1H), 4.50 (t, J = 7.2 Hz, 2H), 4.45 (s, 2H), 4.24 (t, J = 7.2 Hz, 2H), 2.80 (d, J = 15.6 Hz, 2H), 2.70 (d, J = 12.0, 2H), 2.39 - 2.44 (m, 11H), 2.35 (s, 3H).

[0103] Preparation Example 2. Preparation of JP-1366 capsules 5 mg (zaspirone citrate 5 mg)

[0104] 5 mg of azetidin-1-yl {8-[(2,6-dimethylbenzyl)amino]-2,3-dimethylimidazo[1,2-a]pyridin-6-yl}methanone citrate (zastaprazan citrate) obtained according to Preparation Example 1 was mixed with 220.8 mg of D-mannitol, 13.0 mg of croscarmellose sodium, and 1.2 mg of magnesium stearate to obtain a mixture. The mixture was filled into No. 1 hard capsules used as a capsule matrix to prepare a capsule preparation.

[0105] The capsule preparation prepared in this way is referred to as "JP-1366 capsule 5 mg".

[0106] Preparation Example 3. Preparation of JP-1366 tablets 20 mg (zaspirone citrate 20 mg)

[0107] 20 mg of azetidin-1-yl {8-[(2,6-dimethylbenzyl)amino]-2,3-dimethylimidazo[1,2-a]pyridin-6-yl}methanone citrate (zastaprazan citrate) obtained according to Preparation Example 1 was mixed with 133.4 mg of microcrystalline cellulose, 6.4 mg of sodium stearyl fumarate, 40.0 mg of anhydrous lactose, 6.0 mg of croscarmellose sodium, and 4.2 mg of magnesium stearate to obtain a mixture. The mixture was directly compressed to obtain tablets. Then, coating was performed using 8.0 mg of Opadry 03B54445 Pink to prepare film-coated tablets.

[0108] The film-coated tablets prepared as described above are referred to as "JP-1366, 20 mg".

[0109] Preparation Example 4. Preparation of JP-1366 capsules 20 mg (zaspirone citrate 20 mg)

[0110] 20 mg of azetidin-1-yl {8-[(2,6-dimethylbenzyl)amino]-2,3-dimethylimidazo[1,2-a]pyridin-6-yl}methanone citrate (zastaprazan citrate) obtained according to Preparation Example 1 was mixed with 181.4 mg of D-mannitol, 12.4 mg of croscarmellose sodium, and 1.2 mg of magnesium stearate to obtain a mixture. The mixture was filled into No. 1 hard capsules used as a capsule matrix to prepare a capsule preparation.

[0111] The capsule preparation prepared in this way is referred to as "JP-1366 Capsule 20 mg".

[0112] Preparation Example 5. JP-1366 capsules 20 mg placebo

[0113] Zastaprazan capsule 20 mg placebo was prepared in the same manner as Preparation Example 4 except that the main ingredient, zastaprazan citrate, was not used. The capsule preparation prepared in this way is referred to as "JP-1366 Capsule 20 mg Placebo".

[0114] Example 1. In vitro anti-Helicobacter pylori efficacy test - Confirm the bactericidal effect of zaspirone itself

[0115] An experiment was conducted to determine whether zastaprazan has a growth inhibitory or urease activity inhibitory effect on Helicobacter pylori under in vitro experimental conditions.

[0116] 1. Test substances

[0117] A. Drugs

[0118] Table 1 shows the drugs used in the in vitro experiment.

[0119] [Table 1]

[0120] Drugs CAS number Manufacturer Purity Zaspirone (JP-1366) 2133852-18-1 Jeil Pharmaceutical 99.97％ Omeprazole 73590-58-6 TCI >98.0% (HPLC) (T) Amoxicillin trihydrate 61336-70-7 TCI >98.0％(T) Clarithromycin 81103-11-9 TCI >98.0％(T) Nisulaprazan 1902954-60-2 Chemscene 99.58％

[0121] B. Reagents

[0122] Table 2 shows the reagents used in the in vitro experiments.

[0123] [Table 2]

[0124] Reagents CAS number Manufacturer Purity Brain heart infusion 237500 BD (Difco) N / A Muller-Hinton broth 275730 BD (Difco) N / A Bacterial agar 214010 BD (Difco) N / A Lysed horse blood N / A Oxoid N / A IsoVitalex N / A BD (BBL) N / A Horse serum N / A Sigma-Aldrich Hemoglobin, ≤20 mg / dL CampyGen N / A Oxoid N / A

[0125] C. Bacterial Strains

[0126] Helicobacter pylori (ATCC43504): A reference strain used for antibiotic susceptibility testing according to the guidelines of the Clinical and Laboratory Standards Institute (CLSI).

[0127] Helicobacter pylori 26695 (ATCC700329): A strain isolated from a UK patient known to be associated with gastric cancer development. The strain is positive for the cytotoxin-associated gene (CagA) and the vacuolating cytotoxin (VacA), and has a well-known genetic background, so it is mainly used for research purposes.

[0128] Helicobacter pylori SS1 (mouse colonizing strain): A strain known as the Sydney strain. The strain is CagA positive and VacA positive, and has been confirmed to infect and colonize in C57BL / 6 mice. The strain is used for animal testing.

[0129] D. Instruments

[0130] Table 3 shows the instruments used in the in vitro experiments.

[0131] [Table 3]

[0132] Instruments Manufacturer Model 37°C incubator Visionbionex Vision / VS-1203P3V Microcentrifuge Gyrozen 1730R Spectrophotometer BIO-RAD BR170-2525 Microplate reader Molecular Devices SpectraMaxABS

[0133] 2. Culture medium preparation

[0134] A. Culture medium

[0135] Prepare a liquid medium using Brain Heart Infusion (BHI; Difco Inc., Sparks, MD, USA), 7% differentiation medium (horse serum; Sigma CO., LTD., St. Louis, USA), and 0.4% isovitalex (BBL Inc., Sparks, MD, USA), and prepare a solid medium by adding 7% lysed horse blood (Oxoid Inc., Basingstoke, Hants, UK), 0.4% isovitalex, and 1.5 - 2% bacteriological agar (Difco Inc., Sparks, MD, USA) to BHI.

[0136] B. Culture medium for measuring the minimum inhibitory concentration (MIC)

[0137] Prepare a medium using Müller - Hinton broth (MH, Difco Inc., Sparks, MD, USA) [w / 7% differentiation medium (Sigma CO., LTD., St. Louis, USA), 0.4% isovitalex (BBL Inc., Sparks, MD, USA)].

[0138] 3. In vitro efficacy test - MIC and minimum bactericidal concentration (MBC) of zaspirone (JP-1366) against Helicobacter pylori (MBC)

[0139] (1) Method

[0140] According to the CLSI guidelines, perform the antibiotic susceptibility test using the micro - dilution method. Perform a two - fold serial dilution in 12 steps in a 48 - well plate, with the highest concentration of amoxicillin and clarithromycin being 128 μg / mL, and the highest concentration of omeprazole being 256 μg / mL. Inoculate bacteria into wells containing only medium without antibiotics to prepare a positive control, and use wells containing only medium without antibiotics or bacteria as a negative control. Determine the concentration range of the test substances zaspirone (JP - 1366) and nilsurazine through preliminary experiments. Thaw Helicobacter pylori stored in a deep freezer (-70 °C or lower), inoculate it onto BHI agar (w / 7% lysed horse blood and 0.4% isovitalex), culture it at 37 °C for three days under microaerophilic conditions of 5% O2, 10% CO2, and 85% N2 gas, and then sub - culture it on fresh medium. Harvest the cultured bacteria, suspend them in PBS, at 5x10 5Inoculate each well with a concentration of CFU / mL, and then incubate for three days under microaerophilic conditions of 5% O2, 10% CO2, and 85% N2 gas at 37°C. Observe the lowest concentration at which no bacterial growth is detected, and determine it as the MIC. Thereafter, spot 10 μL of the culture solution from each well with different concentrations onto BHI agar without antibiotics (w / 7% lysed horse blood and 0.4% isovitalex), and incubate for three days under microaerophilic conditions at 37°C. The lowest concentration at which no bacterial growth is detected is determined as the MBC.

[0141] (2) Experimental results and analysis

[0142] To determine the MIC, perform visual inspection and measure the absorbance at 600 nm, and confirm growth based on the absorbance of the negative control group containing only the culture medium. To determine the MBC, after determining the MIC, spot 10 μL of the culture solution from each well onto a solid medium without antibiotics or the test substance, and check for bacterial growth.

[0143] Regarding the suitability of the system in this test, quality control is performed based on the MIC value of amoxicillin against the Helicobacter pylori ATCC43504 reference strain.

[0144] All tests are performed in duplicate, and the results are determined through experiments in triplicate.

[0145] Refer to Table 4 below. Due to the low solubility of zasprazan (JP-1366), MIC analysis is difficult (poor solubility, formation of suspensions, interference with visual / absorbance analysis), so the inhibitory effect on Helicobacter pylori strains is confirmed through the MBC index.

[0146] It was confirmed that the test substance zasprazan (JP-1366) showed at least 2-fold and at most 16-fold better inhibitory effects in terms of MBC against three types of Helicobacter pylori strains compared to omeprazole and nisulazan.

[0147] [Table 4]

[0148]

[0149] Example 2. Anti-Helicobacter pylori efficacy test in patients with erosive gastroesophageal reflux disease (phase 2 clinical trial) - Confirm the bactericidal effect of zaspirone itself

[0150] 1. Subject selection

[0151] Helicobacter pylori test

[0152] Helicobacter pylori testing is performed by 13C-urea breath test (13C-UBT), gastroscopy (Campylobacter-like organism test, CLO), or tissue biopsy to confirm Helicobacter pylori positive / negative. The test is performed at the screening visit (Visit 1) or randomization visit (Visit 2), and the results from the same institution within 29 days prior to the randomization visit (Visit 2) can be substituted.

[0153] Inclusion criteria

[0154] Unless otherwise specified, the subjects need to meet all of the following inclusion criteria to participate in this clinical trial.

[0155] 1) Adult men and women aged 19 to 75 years as of the date of written consent

[0156] 2) Subjects who have experienced heartburn or acid reflux symptoms within seven days prior to the screening visit (Visit 1)

[0157]

[0158] 3) Subjects who have been diagnosed with grade A or higher erosive gastroesophageal reflux disease according to the Los Angeles classification by upper gastrointestinal endoscopy at the same institution within 29 days prior to the randomization visit (Visit 2)

[0159] LA grade Criteria A One or more mucosal breaks in the esophagus with a length < 5 mm B One or more mucosal breaks in the esophagus with a length of ≥5 mm C Continuous esophageal mucosal breaks, but involving less than or equal to 75% of the esophageal circumference D Esophageal mucosal breaks involving at least 75% of the esophageal circumference

[0160] 4) Subjects who are able to complete the questionnaire and subject diary

[0161] 5) Subjects who have voluntarily given written consent to participate in this clinical trial

[0162] Exclusion criteria

[0163] Subjects who meet any of the following criteria will be excluded from this clinical trial.

[0164] 1) Excluded diseases

[0165] (1) Subjects who have been determined to have Barrett's esophagus more than 3 cm or significant dysplastic changes by upper gastrointestinal endoscopy at the screening visit (Visit 1)

[0166] (2) Eosinophilic esophagitis (subjects with negative results on esophageal biopsy can be included)

[0167] (3) Subjects with primary esophageal motility disorders or esophageal strictures

[0168] (4) Gastroesophageal varices

[0169] (5) Subjects with gastrointestinal (GI) bleeding or other abnormal bleeding identified by upper gastrointestinal endoscopy at the screening visit (Visit 1)

[0170] (6) Subjects who have undergone surgery for acid secretion inhibition or surgery on the stomach or esophagus. However, subjects meeting the following criteria may be included.

[0171] ① Appendectomy, cholecystectomy, polypectomy

[0172] ② Endoscopic resection of malignant tumors or endoscopic resection of early gastric cancer (endoscopic mucosal resection (EMR) and endoscopic submucosal dissection (ESD)), which has healed and has not recurred for more than five years since the date of diagnosis

[0173] (7) Subjects with active duodenal ulcer, gastric ulcer, or pancreatitis

[0174] (8) Zollinger-Ellison syndrome

[0175] (9) Subjects with irritable bowel syndrome (IBS) or inflammatory bowel disease (IBD)

[0176] (10) Subjects with warning symptoms (such as dysphagia, severe dysphagia, bleeding, weight loss, anemia, bloody stools) who are suspected of having a gastrointestinal malignancy by the investigator's judgment (however, subjects with a negative result on upper gastrointestinal endoscopy confirming the presence of a tumor may be included)

[0177] 2) Past medical history and comorbidities

[0178] (1) Subjects with clinically significant liver, kidney, nervous system, respiratory system, endocrine system, hematological oncology, cardiovascular system, or urinary system diseases

[0179] (2) Subjects with a history of malignancy within five years before the screening visit (Visit 1) (however, subjects who have been cured and have not recurred for more than five years since the date of diagnosis may be included, but subjects with a history of digestive system malignancy and tumor resection (such as gastrectomy, colectomy, etc.) may not be included, except in cases (1)-(6))

[0180] (3) Subjects with a history of alcohol or drug abuse within one year before the screening visit (Visit 1)

[0181] (4) Subjects with a confirmed history of human immunodeficiency virus (HIV)

[0182] (5) Subjects diagnosed with mental illnesses (such as schizophrenia, dementia) that may affect the conduct of this clinical trial

[0183] (3) Laboratory test results (at screening visit)

[0184] (1) Subjects with serum AST, ALT, ALP, γ-GT, or total bilirubin levels more than twice the upper limit of normal

[0185] (2) Subjects with serum BUN or serum creatinine levels more than twice the upper limit of normal

[0186] (3) Active hepatitis B or C (subjects with undetectable virus can be included)

[0187] 4) Drug intake / treatment history

[0188] (1) Subjects who are taking medications that are contraindicated for concomitant use or are expected to take them during the clinical trial

[0189] 5) Allergy (hypersensitivity) history

[0190] (1) Patients with allergies and a history of allergies to the clinical trial drug, components of Nexium tablets (esomeprazole), benzimidazoles, penicillin antibiotics, and macrolide antibiotics

[0191] 6) Others

[0192] (1) Pregnant women, lactating women, or women with a positive pregnancy test, or fertile (reproductive-age) women or men planning to become pregnant during this clinical trial

[0193] (2) Female partners of female and male subjects of reproductive potential who have not undergone infertility surgery and do not agree to use the following contraceptive methods during the clinical trial

[0194] (3) Subjects who have received other clinical trial drugs within one month before the screening visit (Visit 1)

[0195] (4) In addition to the above, subjects determined by the investigator to be unsuitable for participation in this clinical trial

[0196] 2. Clinical trial method

[0197] This clinical trial was a randomized, double-blind, active-controlled, multi-center, Phase 2 clinical trial for patients with erosive gastroesophageal reflux disease. After the subjects gave written consent to participate in the clinical trial, screening tests were conducted. When the subjects took medications that might affect the gastric mucosa during screening, upper gastrointestinal endoscopy was performed after a 2-week drug-free period (4-week drug-free period for potassium-competitive acid blockers (P-CABs) or proton pump inhibitors (PPIs)). According to the results of the screening tests, the subjects who met the inclusion / exclusion criteria were randomly assigned to Test Group 1, Test Group 2, or the control group at a ratio of 1:1:1. At this time, the subjects were stratified by grade (A, B, C, D) according to the LA classification system classified by upper gastrointestinal endoscopy. The randomly assigned subjects took the clinical trial drug once a day for four weeks according to the assigned administration group. Four weeks after administering the clinical trial drug, the subjects visited the clinical trial institution for upper gastrointestinal endoscopy to evaluate whether they were cured. The cured subjects had a safety follow-up (F / U) two weeks later, and the uncured subjects were administered the clinical trial drug for another four weeks (a total of eight weeks) and visited the clinical trial institution for upper gastrointestinal endoscopy. Regardless of whether they were cured, a safety F / U was conducted two weeks later.

[0198] 3. Administration dose, administration period, and administration method

[0199] A. Administration dose

[0200] - Test Drug 1: JP-1366, 5 mg, two capsules (zasprazan citrate 10 mg)

[0201] - Test Drug 2: JP-1366, 20 mg, one capsule (zasprazan citrate 20 mg)

[0202] - Control Drug: Esomeprazole Magnesium Trihydrate 44.5 mg (40 mg as esomeprazole, 1 tablet of Nexium 40 mg)

[0203] B. Administration period

[0204] The trial period for each subject: up to 14 weeks

[0205] - Screening period: 15 days or 29 days

[0206] - Administration period: four weeks (or eight weeks)

[0207] - Safety F / U period: two weeks after the last administration of the clinical trial drug

[0208] C. Administration Method

[0209] According to the randomly assigned treatment group, starting from the day when the investigational clinical drug is provided, the subject orally administers the investigational clinical drug once a day at regular intervals in a fasting state for four weeks (or eight weeks*). *Only subjects whose mucosal lesions do not heal four weeks after the administration of the investigational clinical drug are given an additional four-week treatment period.

[0210] 4. Efficacy Endpoint

[0211] A. Primary Efficacy Endpoint:

[0212] Cumulative healing rate of mucosal lesions (%) until week 8 after administration of the investigational clinical drug

[0213] *Erosion is found to have recovered to normal mucosa by upper gastrointestinal endoscopy

[0214] B. Secondary Efficacy Endpoint:

[0215] ① Healing rate of mucosal lesions (%) until week 4 after administration of the investigational clinical drug

[0216] ② Symptom assessment through the subject's diary

[0217] A. Assessment of symptoms (heartburn, acid reflux, heartburn / acid reflux) within 24 hours after administration of the investigational clinical drug

[0218] B. Assessment of symptoms (heartburn, acid reflux, heartburn / acid reflux) seven days after administration of the investigational clinical drug

[0219] C. Number of days to achieve complete remission (CR, disappearance of heartburn and acid reflux for seven days) after administration of the investigational clinical drug

[0220] D. Proportion of asymptomatic days (heartburn, acid reflux, heartburn / acid reflux) at week 4 and week 8 after administration of the investigational clinical drug (daytime, nighttime, daytime / nighttime)

[0221] ③ Gastroesophageal reflux disease symptom assessment (Reflux Disease Questionnaire (RDQ)): Changes in the frequency and severity of the main symptoms at week 4 and week 8 after administration of the investigational clinical drug compared to baseline

[0222] ④ Quality of life assessment (Gastroesophageal Reflux Disease - Health-Related Quality of Life (GERD-HRQL)): Changes in the total score at week 4 and week 8 after administration of the investigational clinical drug compared to baseline

[0223] 5. Safety Endpoint

[0224] (1) Adverse reactions

[0225] (2) Vital signs

[0226] (3) Laboratory tests

[0227] (4) Electrocardiogram (12 - lead ECG)

[0228] (5) Physical examination

[0229] 6. Statistical Analysis Method

[0230] A. Primary Efficacy Endpoint

[0231] Cumulative mucosal lesion healing rate (%) from the administration of the clinical trial drug until week 8

[0232] The proportion of subjects whose erosions have returned to normal mucosa from the first administration of the clinical trial drug until week 8 and the two - sided 95% confidence intervals are presented by treatment group. To compare each test group with the control groups (JP - 1366 10 mg vs Nexium tablet 40 mg, JP - 1366 20 mg vs Nexium tablet 40 mg), the Cochran - Mantel - Haenszel method adjusted for the baseline LA grade (A, B, C, D) as a stratification factor is used to obtain the lower limit of each two - sided 95% confidence interval.

[0233] B. Secondary Efficacy Endpoint

[0234] ① Mucosal lesion healing rate (%) from the administration of the clinical trial drug until week 4

[0235] The proportion of subjects whose erosions have returned to normal mucosa from the first administration of the clinical trial drug until week 4 and the two - sided 95% confidence intervals are presented by treatment group. To compare each test group with the control groups (JP - 1366 10 mg vs Nexium tablet 40 mg, JP - 1366 20 mg vs Nexium tablet 40 mg), the Cochran - Mantel - Haenszel method adjusted for the baseline LA grade (A, B, C, D) as a stratification factor is used to obtain the lower limit of each two - sided 95% confidence interval.

[0236] ② Symptom assessment via subject diaries

[0237] A. Assessment of symptoms (heartburn, acid reflux, heartburn / acid reflux) within 24 hours after administration of the clinical trial drug: Using an analysis of covariance (ANCOVA) model with the baseline LA grade (A, B, C, D) as a covariate, the comparison of the mean assessment of symptoms (heartburn, acid reflux, heartburn / acid reflux) within 24 hours after the first administration of the clinical trial drug between each test group and the control groups is analyzed.

[0238] B. Assessment of symptoms (heartburn, acid reflux, heartburn / acid reflux) seven days after administration of the investigational drug: Using an ANCOVA model with baseline LA grade (A, B, C, D) as a covariate, analyze the comparison of the mean assessment of symptoms (heartburn, acid reflux, heartburn / acid reflux) between each test group and the control group seven days after the first administration of the investigational drug.

[0239] C. Number of days to achieve CR (absence of heartburn and acid reflux for seven days) after administration of the investigational drug: When the symptom score is 0 for seven consecutive days after the first administration of the investigational drug, it is defined as CR (absence of heartburn and acid reflux for seven days), and the event is based on the first CR. When there is no CR, the case is considered censored. Kaplan-Meier curves and the median and its two-sided 95% confidence intervals are presented by treatment group, and a stratified log-rank test adjusted for baseline LA grade as a stratification factor is used to analyze the comparison between each test group and the control group.

[0240] D. Proportion of days without symptoms (heartburn, acid reflux, heartburn / acid reflux) at 4 and 8 weeks after administration of the investigational drug (daytime, nighttime, daytime / nighttime): Using an ANCOVA model with baseline LA grade (A, B, C, D) as a covariate, analyze the comparison of the proportion of days without symptoms (heartburn, acid reflux, heartburn and acid reflux) during the day, at night, and during the day and night between each test group and the control group at 4 and 8 weeks after the first administration of the investigational drug.

[0241] ③ Symptom assessment of gastroesophageal reflux disease (RDQ)

[0242] Using an ANCOVA model with baseline LA grade (A, B, C, D) as a covariate, analyze the comparison of the changes in the frequency and severity of the main symptoms between each test group and the control group at 4 and 8 weeks after administration compared to the baseline. The same statistical analysis as above is performed on the changes in the final results (excluding the baseline) compared to the baseline.

[0243] ④ Quality of life assessment (GERD-HRQL)

[0244] Using an ANCOVA model with baseline LA grade (A, B, C, D) as a covariate, analyze the comparison of the changes in the total quality of life scores between each test group and the control group at 4 and 8 weeks after administration compared to the baseline. The same statistical analysis is performed on the changes in the final results (excluding the baseline) compared to the baseline.

[0245] C. Safety Endpoint

[0246] For subjects who experienced at least one adverse reaction (or drug adverse reaction), the occurrence times and incidence rates and their two-sided 95% confidence intervals were presented by treatment group, and a chi-square test or Fisher's exact test was performed to test for differences in the incidence rates of adverse reactions (or drug adverse reactions) between the control group and each test group. In addition, the data were coded and presented as System Organ Class (SOC) and Preferred Term (PT) using the latest version of the Medical Dictionary for Regulatory Activities (MedDRA). Serious adverse reactions, adverse reactions that led to interruption of the administration of the clinical trial drug, and adverse reactions that led to death were also summarized using the latest version of MedDRA as SOC and PT. For laboratory tests, vital signs, and electrocardiograms (12-lead ECGs), descriptive statistics (number of subjects, mean, standard deviation, median, minimum, maximum), changes at each visit, and changes relative to baseline were presented as continuous data, and contingency tables were created for categorical data. For laboratory tests and electrocardiograms (12-lead ECGs), changes from baseline after administration of the clinical trial drug were summarized in the form of a transition table at each assessment time point, categorized as normal or clinically insignificant abnormalities (normal or abnormal NCS) and clinically significant abnormalities (abnormal CS). For physical examinations, a list of subjects who were normal or had clinically insignificant abnormalities (abnormal NCS) at baseline but changed to clinically significant abnormalities (abnormal CS) after administration of the clinical trial drug was presented.

[0247] 7. Results of Phase 2 Clinical Trial

[0248] A. Confirmation of Symptom Improvement

[0249] In the assessment of all symptoms (heartburn, acid reflux, heartburn / acid reflux) within 24 hours after administration of the clinical trial drug and for seven consecutive days, it was confirmed that all symptoms tended to improve within 24 hours after administration of Test Drugs 1 and 2, which were used as clinical trial drugs, and for seven consecutive days.

[0250] B. Confirmation of Healing Rate

[0251] (1) Helicobacter pylori Positive Group (Infection Group)

[0252] The JP-1366 20 mg (zaspirone 20 mg) treatment group showed a 100% cure rate at both Week 4 and Week 8, while Nexium 40 mg showed an 87.5% effect (see Table 5).

[0253] [Table 5]

[0254]

[0255] (2) Helicobacter pylori Negative Group

[0256] It was confirmed that the JP-1366 20 mg (zaspirone 20 mg) administration group showed the same effect as the Nexium 40 mg group (see Table 6). From this, it was clinically confirmed that zaspirone can show the same effect as the control Nexium 40 mg even at a significantly low dose.

[0257] [Table 6]

[0258]

[0259]

[0260] (3) Overall Healing Rate

[0261] In terms of the overall cure rate, considering the results of the Helicobacter pylori positive group (infected group) and the Helicobacter pylori negative group, at the 4th week after administration of the clinical trial drug, the proportion of subjects with complete mucosal defect healing was 93.75% (45 / 48 people) in the JP-1366 10 mg group, 91.49% (43 / 47 people) in the JP-1366 20 mg group, and 89.80% (44 / 49 people) in the esomeprazole 40 mg group.

[0262] (4) Conclusion

[0263] Nexium 40 mg showed a higher erosion cure rate in Helicobacter pylori negative patients than in Helicobacter pylori positive patients, but the pharmaceutical composition of the present invention showed a therapeutic effect on gastroesophageal reflux disease and / or peptic ulcer regardless of Helicobacter pylori infection. On the contrary, it showed a tendency to show a higher effect in patients infected with Helicobacter pylori. This is due to the antibacterial effect of zaspirone itself contained in the pharmaceutical composition according to the present invention.

[0264] Example 3. Anti-Helicobacter pylori Efficacy Test in Patients with Erosive Gastroesophageal Reflux Disease (Phase 3 Clinical Trial)- Confirmation of Bacterial Eradication Effect of Zastaprazan Itself

[0265] A phase 3 clinical trial was conducted by increasing the number of subjects. Specifically, according to the randomly assigned administration group, the clinical trial drug was orally administered once a day at regular intervals when possible, regardless of whether it was during a meal, for a total of four weeks, and the cumulative healing rate of mucosal defects was confirmed in Helicobacter pylori positive subjects.

[0266]

[0267] Per Protocol Set (PPS)

[0268] As a result of the PPS analysis (which is the primary analysis group of this clinical trial), the number of subjects tested positive for Helicobacter pylori was 26 in the test group and 22 in the control group. The proportion of subjects with endoscopic mucosal break healing at 4 weeks after administration of the clinical trial drug was 100.00% (26 / 26 subjects) in the test group and 81.82% (18 / 22 subjects) in the control group, showing a statistically significant difference between the two administration groups (p = 0.0214).

[0269] Full Analysis Set (FAS)

[0270] According to the FAS analysis results, the number of subjects tested positive for Helicobacter pylori was 28 in the test group and 25 in the control group. The percentage of subjects with endoscopic mucosal break healing at 4 weeks after administration of the clinical trial drug was 96.43% (27 / 28 subjects) in the test group and 80.00% (20 / 25 subjects) in the control group. The test group showed excellent effects with significant differences compared to the control group at 4 weeks.

[0271] [Table 7]

[0272] JP-1366 20mg Esomeprazole 40mg Number of Subjects 26 22 Number of Cured Subjects, n(%) 26(100.00) 18(81.82)

[0273] [Table 8]

[0274] JP-1366 20mg Esomeprazole 40mg Number of Subjects 28 25 Number of Cured Subjects, n(%) 27(96.43) 20(80.00)

[0275] Example 4. Evaluation of Drug Interaction between Zastaprazan (JP-1366), Amoxicillin and Clarithromycin in Healthy Adult Volunteers 1. Purpose of Clinical Trial

[0276] 2. Test Drugs and Administration Period

[0277] The purpose of this clinical trial was to evaluate the effects of co - administration of amoxicillin and clarithromycin on the PK, safety, and tolerability of JP - 1366, and the effects of JP - 1366 on the PK, safety, and tolerability of amoxicillin and clarithromycin in healthy adult volunteers. The target number of subjects was 24, and the clinical trial was conducted in healthy adults in a randomized, open - label, three - way, repeated - dose, crossover design.

[0278] A. Test Drug 1: JP-1366 Capsule 20mg

[0279] B. Test Drug 2: Kymoxin Capsule 500mg

[0280] Manufacturer: Jeil Pharmaceutical

[0281] Dose: JP - 1366 20mg

[0282] Route of administration: Oral

[0283] C. Test Drug 3: Clarithromycin (Klaricid) Film-Coated Tablet 500mg

[0284] Manufacturer: Yuhan Corporation CO., LTD.

[0285] Dosage: 500 mg of amoxicillin hydrate

[0286] Route of administration: Oral

[0287] D. Administration Period

[0288] Dosage: 500 mg of clarithromycin

[0289] Manufacturer: Abbott Korea CO., LTD.

[0290] Route of administration: Oral

[0291] 3. Selection and Classification of Clinical Trial Subjects

[0292] Twice a day, for 5 days each period

[0293] 4. Results

[0294] A total of 24 subjects participated in this clinical trial and received the clinical trial drug, and all subjects completed the entire trial process and all visits according to the clinical trial protocol. Therefore, pharmacokinetic evaluations and safety evaluations were performed on 24 subjects. Statistical analyses of PK were performed for Part A and Part B.

[0295] Part A: 20 mg of JP-1366, 1 capsule, twice a day, repeated for 5 days (T1) or 1 capsule of JP-1366 + 2 capsules of 500 mg of amoxicillin + 1 tablet of 500 mg of clarithromycin, twice a day, repeated for 5 days (T3)

[0296] Part B: 2 capsules of 500 mg of amoxicillin + 1 tablet of 500 mg of clarithromycin, twice a day, repeated for 5 days (T2) or 20 mg of JP-1366, 1 capsule + 2 capsules of 500 mg of amoxicillin + 1 tablet of 500 mg of clarithromycin, twice a day, repeated for 5 days (T3)

[0297] Example 5. In Vitro Anti-Helicobacter pylori Efficacy Test - Confirmation of Bacterial Eradication Effect of Co-Administration of Zastaprazan (JP-1366) and Clarithromycin

[0298] [Part A]

[0299] The results of the pharmacokinetic evaluation of JP-1366 are shown below.

[0300] [Table 9]

[0301]

[0302] (In Table 9, T1 represents 1 capsule of JP-1366 20 mg, and T3 represents 1 capsule of JP-1366 20 mg + 2 capsules of amoxicillin 500 mg + 1 tablet of clarithromycin 500 mg.)

[0303] In Part A, 1 capsule of JP-1366 20 mg was administered twice a day for five days (T1) or 1 capsule of JP-1366 + 2 capsules of amoxicillin 500 mg + 1 tablet of clarithromycin 500 mg was administered twice a day for 5 days (T3). The C max,ss and AUC T were set as the primary endpoints, and the 90% confidence interval of the geometric mean ratio (T3 / T1) was calculated to evaluate the effect of co-administration with amoxicillin and clarithromycin on the PK of JP-1366. As a result of the PK evaluation, the C max,ss and AUC T of JP-1366 exceeded the upper limit of the 90% confidence interval of the geometric mean ratio (T3 / T1) of 0.8 to 1.25. Therefore, when T3 was administered, the C max,ss was approximately 1.8 times higher, and the AUC T was approximately 2.4 times higher than when T1 was administered.

[0304] [Part B]

[0305] The PK evaluation results of amoxicillin are shown below.

[0306] [Table 10]

[0307]

[0308] (In Table 10, T2 represents 2 capsules of amoxicillin 500 mg + 1 tablet of clarithromycin 500 mg, and T3 represents 1 capsule of JP-1366 20 mg + 2 capsules of amoxicillin 500 mg + 1 tablet of clarithromycin 500 mg.)

[0309] In Part B, 2 capsules of amoxicillin 500 mg + 1 tablet of clarithromycin 500 mg were administered twice a day for five days (T2), or 1 capsule of JP-1366 20 mg + 2 capsules of amoxicillin 500 mg + 1 tablet of clarithromycin 500 mg were administered twice a day for five days (T3), and the C max,ss and AUC TSet as the primary endpoint, and the 90% confidence interval of the geometric mean ratio (T3 / T2) was calculated to evaluate the effect of co - administration with amoxicillin and clarithromycin on the PK of JP - 1366. As a result of the PK assessment, the AUC of amoxicillin T was within the 90% confidence interval of the geometric mean ratio (T3 / T2) of 0.8 to 1.25, but the C of amoxicillin max,ss was outside the range of 0.8 to 1.25, indicating that when T3 was administered, the C of amoxicillin max,ss was approximately 20% lower than when T2 was administered.

[0310] The PK assessment results of clarithromycin are shown below.

[0311] [Table 11]

[0312]

[0313] The C of clarithromycin max,ss was within the 90% confidence interval of the geometric mean ratio (T3 / T2) of 0.8 to 1.25, but the AUC of clarithromycin T was outside the range of 0.8 to 1.25, indicating that when T3 was administered, the AUC of clarithromycin T was approximately 14% higher than when T2 was administered. When comparing the co - administration of amoxicillin, clarithromycin, and JP - 1366 with the administration of amoxicillin and clarithromycin alone, the difference in drug exposure was within 2 - fold, indicating no clinically significant PK drug interactions.

[0314] 1. Test Purpose 2. Test Substances

[0315] A. Drugs

[0316] The purpose of this test was to confirm the antibacterial activity of zaspirone (JP - 1366), which is different from omeprazole or nisulazan, in the first antibacterial activity test. Before confirming the enhancing effect of zaspirone (JP - 1366) on the antibacterial activity of clarithromycin used in the triple - drug regimen, this additional study was conducted under in vitro experimental conditions to confirm the antibacterial activity synergy against sensitive strains when zaspirone (JP - 1366) and clarithromycin were co - administered as a dual - drug regimen.

[0317] Drug

[0318] CAS Number

[0319] The drugs used in the in vitro experiment are shown in Table 12 below.

[0320] [Table 12]

[0321] Manufacturer Purity Zastaprazan (JP-1366) Jeil Pharmaceutical Clarithromycin 2133852-18-1 TCI 99.97％ B. Reagents 81103-11-9 Reagent >98.0％(T)

[0322] CAS Number

[0323] The reagents used in the in vitro experiments are shown in Table 13 below.

[0324] [Table 13]

[0325] Manufacturer Purity Brain Heart Infusion BD (Difco) N / A 237500 Muller-Hinton Broth BD (Difco) N / A 275730 Bacterial Agar BD (Difco) N / A 214010 Lysed Horse Blood N / A Oxoid N / A Isovitalex N / A BD (BBL) N / A Horse Serum N / A Sigma-Aldrich Hemoglobin, ≤20mg / dL CampyGen N / A Oxoid N / A C. Bacterial Strains D. Instruments

[0326] Instrument

[0327] Helicobacter pylori SS1 (mouse colonization strain): known as the Sydney strain, which is CagA positive and VacA positive, and has been confirmed to colonize mice C57BL / 6 and is used in animal experiments.

[0328] Manufacturer

[0329] The instruments used in the in vitro experiments are shown in Table 14 below.

[0330] [Table 14]

[0331] Model 37°C Incubator Visionbionex Vision / VS-1203P3V Microcentrifuge Gyrozen Spectrophotometer BIO-RAD 1730R BR170-2525 Microplate Reader Molecular Devices SpectraMaxABS 3. Preparation of Culture Media and Test Substances A. Culture Media

[0332] B. Culture Media for Measuring MIC

[0333] C. Preparation of Test Substances

[0334] The culture medium was prepared by adding 7% lysed horse blood (Oxoid Ltd, Basingstoke, Hants, UK), 0.4% IsoVitalex, and 1.5% to 2% bacteriological agar (Difco, Sparks, MD, USA) to BHI.

[0335] 4. In Vitro Efficacy Test

[0336] Mueller Hinton broth (MH, Difco, Sparks, MD, USA) [w / 7% horse serum (Sigma Co. St. Louis, USA), 0.4% IsoVitalex (BBL, Sparks, MD, USA)] was used, and for the purpose of observing synergy, JP-1366 at 1 / 2 of the MIC value of zaspirone (JP-1366) was added to the MH culture medium prepared above, and the resulting mixture was used as the basal medium for MIC measurement.

[0337] A. Measurement of MIC of Zastaprazan (JP-1366) against Helicobacter pylori SS1

[0338] Zasaprazan (JP-1366) is provided as a product prepared at a concentration of 10 mM in dimethyl sulfoxide (DMSO) solvent, and clarithromycin is prepared at a concentration of 5 mg / mL in DMSO solvent.

[0339] B. Efficacy of Zastaprazan (JP-1366) in Enhancing the Antibacterial Activity of Clarithromycin against Helicobacter pylori

[0340] 5. Experimental Results

[0341] An antibiotic susceptibility test was performed to measure the MIC by preparing a solid medium containing JP-1366 in a 6-well plate using MH agar (7% horse serum, 0.4% IsoVitalex, 1.5% to 2% agar) at the highest concentration of JP-1366 of 32 μg / mL (which is the maximum MBC value), and performing 6-step dilutions with a maximum culture medium volume of 2 mL. Helicobacter pylori stored in a deep freezer (-70 °C or lower) was inoculated onto BHI agar (w / 7% lysed horse blood, 0.4% IsoVitalex), cultured at 37 °C under microaerophilic conditions for 3 days (Oxoid CampyGene gas pk), and then subcultured on fresh medium. The cultured bacteria were harvested, suspended in PBS, and then suspended at 0.5 McFarland unit (1.5x 10 8 CFU / mL). Then, 2 μL was inoculated onto the solid medium and cultured under microaerophilic conditions for three days. The lowest concentration at which no growth was visually observed was determined as the MIC.

[0342] ​

[0343] According to the CLSI guidelines, the antibiotic susceptibility test was performed by the microdilution method.

[0344] Antibiotic susceptibility tests were performed using MH broth (7% horse serum, 0.4% IsoVitalex), and at this time, a susceptibility test medium for zaspirone (JP-1366) was prepared. The tests were carried out in 48-well plates with a final medium volume of 200 μL, and the concentration of zaspirone (JP-1366) was half of the MIC value of zaspirone (JP-1366). Using the medium prepared in this way, clarithromycin was serially diluted 2-fold in 16 steps with a maximum concentration of 16 μg / mL. The wells containing only the medium without antibiotics were used as the negative control group, and the wells inoculated only with bacteria in the medium without antibiotics were used as the positive control group. Helicobacter pylori stored in a deep freezer (-70 °C or lower) was inoculated onto BHI agar (w / 7% lysed horse blood, 0.4% IsoVitalex), cultured for three days at 37 °C under microaerophilic conditions (Oxoid CampyGene gaspk), and then subcultured on fresh medium. The cultured bacteria were harvested, suspended in PBS, and inoculated into each well at a concentration of 5x10 5 CFU / mL. Thereafter, the bacteria were cultured for three days at 37 °C under microaerophilic conditions of 5% O2, 10% CO2, and 85% N2 gas. The lowest concentration of the sample where no bacterial growth was observed was determined as the MIC. Thereafter, 10 μL of the culture solution from each well of different concentrations for measuring the MIC was spotted onto BHI agar without antibiotics (7% lysed horse blood, 0.4% IsoVitalex) and incubated for three days at 37 °C under microaerophilic conditions (Oxoid CampyGene gas pk). The lowest concentration where no bacterial growth was observed was determined as the minimum bactericidal concentration (MBC).

[0345] ​

[0346] In this experiment, the efficacy of JP-1366 in enhancing the antibacterial activity of clarithromycin against Helicobacter pylori was determined by one of the following methods.

[0347] 1) When the drug is co-treated with clarithromycin, the MIC (or MBC) of clarithromycin decreases.

[0348] 2) When the drug is co-treated with clarithromycin at a specific concentration that inhibits the growth of antibiotic-resistant strains of bacteria, the proportion (%) of strains showing cell growth inhibition increases.

[0349] Referring to Table 15, when clarithromycin was added to the medium containing zaspirone (JP1366) at a concentration of 1 / 2 MIC, the MBC decreased by approximately 1 / 2. To more clearly confirm the effect, additional experiments were conducted on resistant strains (see Example 6 below).

[0350] [Table 15]

[0351]

[0352] Example 6. In vitro efficacy test against Helicobacter pylori - Confirmation of the bactericidal effect of co - administration of zasprazan (JP - 1366), amoxicillin, and clarithromycin

[0353] 1. Test objective

[0354] The purpose of this experiment was to confirm the bactericidal eradication synergy of the triple-drug regimen of zaspirone (JP-1366), amoxicillin, and clarithromycin against resistant strains under in vitro experimental conditions.

[0355] 2. Test substances

[0356] The bacterial strains used in this experiment were as follows.

[0357] - Helicobacter pylori ATCC43503: The standard strain for antibiotic susceptibility testing

[0358] - Clinically isolated antibiotic-resistant strains: Provided by the Gyeongsang National University Hospital Uncultured Pathogen Resource Bank

[0359] [Table 16]

[0360] Helicobacter pylori KBN12P07353 Helicobacter pylori KBN12P08132 Helicobacter pylori KBN12P08133 Helicobacter pylori KBN12P08134 Helicobacter pylori KBN12P08135 Helicobacter pylori KBN12P08136 Helicobacter pylori KBN12P08137 Helicobacter pylori KBN12P08138 Helicobacter pylori KBN12P08182 Helicobacter pylori KBN12P08204

[0361] 3. Preparation of culture medium and test substances

[0362] The preparation of the culture medium, MIC measurement medium, and test substances was the same as described in Example 5.

[0363] 3. In vitro efficacy test

[0364] A. Test for confirming the MIC and MBC of zasprazan (JP - 1366) against 10 strains

[0365] According to the CLSI guidelines, the antibiotic susceptibility test was performed by the microdilution method.

[0366] Amoxicillin and clarithromycin were provided at 128 μg / mL, amoxicillin + clarithromycin were mixed at a 1:1 ratio to 64 μg / mL, and JP-1366, omeprazole, and gefarnate were subjected to 12 steps of 2-fold dilution with a maximum concentration of 256 μg / mL. Tubes containing only medium without antibiotics were inoculated with bacteria and used as the positive control group, and tubes containing only medium without antibiotics and bacteria were used as the negative control group. Helicobacter pylori stored in a -70 °C deep freezer was thawed and inoculated onto BHI agar (w / 7% lysed horse blood, 0.4% IsoVitalex), cultured at 37 °C for three days under microaerophilic conditions of 5% O2, 10% CO2, and 85% N2 gas, and subcultured on fresh medium. The cultured bacteria were harvested, suspended in sterile PBS, and inoculated into tubes with different drug concentrations at a concentration of 5x10 5 CFU / mL, and cultured at 37 °C for three days under microaerophilic conditions of 5% O2, 10% CO2, and 85% N2 gas. The lowest concentration at which no bacterial growth was observed was determined as the MIC. Thereafter, 10 μL of the culture solution from each tube of different concentrations was spotted onto BHI agar without antibiotics (w / 7% lysed horse blood, 0.4% IsoVitalex) and incubated at 37 °C under microaerophilic conditions for three days. The lowest concentration at which no bacterial growth was observed was determined as the minimum bactericidal concentration (MBC).

[0367] B. Efficacy of zasprazan (JP - 1366) in enhancing the antibacterial efficacy of amoxicillin and clarithromycin against Helicobacter pylori efficacy

[0368] Antibiotic susceptibility testing was performed by the microdilution method according to CLSI guidelines.

[0369] Antibiotic susceptibility tests were performed using MH broth (7% horse serum, 0.4% IsoVitalex), and three types of susceptibility test media containing zaspirone (JP-1366), omeprazole, and nisulaprazan were prepared. The experiments were conducted in 48-well plates with a final medium volume of 200 μL, and the concentrations of zaspirone (JP-1366), nisulaprazan, and omeprazole contained in the medium were set to be the same as the MIC values of zaspirone (JP-1366) against each clinical isolate (when it was difficult to confirm the MIC, the highest concentration at which bacteria survived after measurement using MBC was used). Using the medium prepared in this way, amoxicillin and clarithromycin (mixed in a 1:1 ratio) were serially diluted 2-fold for 12 steps, with a maximum concentration of 64 μg / mL. The wells containing only the medium without antibiotics were used as the negative control group, and the wells inoculated only with bacteria in the medium without antibiotics were used as the positive control group. Helicobacter pylori was inoculated onto BHI agar (w / 7% lysed horse blood, 0.4% IsoVitalex) and cultured under microaerobic conditions at 37 °C for three days (Oxoid CampyGene gas pk), and then subcultured on fresh medium. The cultured bacteria were harvested, suspended in PBS, and inoculated into each well at a concentration of 5x10 5 CFU / mL. Thereafter, the bacteria were cultured under microaerobic conditions of 5% O2, 10% CO2, and 85% N2 gas at 37 °C for three days. The lowest sample concentration at which no bacterial growth was observed was determined as the MIC. Thereafter, 10 μL of the culture solution from each well at different concentrations measured for the MIC was spotted onto BHI agar (7% lysed horse blood, 0.4% IsoVitalex) without antibiotics and incubated under microaerobic conditions (Oxoid CampyGene gas pk) at 37 °C for three days. The lowest concentration at which no bacterial growth was observed was determined as the MBC. At this time, quality control of the amoxicillin susceptibility test for HP43504 was performed.

[0370] C. Case of antibiotic combination

[0371] As shown in Table 17 below, a total of three antibiotic combinations were subjected to susceptibility tests for amoxicillin and clarithromycin against 10 strains.

[0372] [Table 17]

[0373] Type of culture medium for MIC / MBC measurement Antibiotics for MIC / MBC measurement JP - 1366 medium Amoxicillin + clarithromycin Omeprazole medium Amoxicillin + clarithromycin Fisulazan - free medium Amoxicillin + clarithromycin

[0374] In this experiment, the efficacy of the drugs (zaspirone (JP-1366), omeprazole, nisulaprazan) in enhancing the antibacterial activity of amoxicillin and clarithromycin against Helicobacter pylori was judged by one of the following methods.

[0375] 1) When the drug is treated together with amoxicillin and clarithromycin, the MIC (or MBC) of amoxicillin and clarithromycin decreases.

[0376] 2) When the drug is treated together with specific concentrations of amoxicillin and clarithromycin for the growth of antibiotic-resistant strain bacteria, the proportion (%) of strains with inhibited cell growth is increased.

[0377] Although specific parts of the present invention have been described in detail above, it is obvious to those skilled in the art that these specific descriptions are merely preferred embodiments, and the scope of the present invention is not limited thereto. Therefore, the actual scope of the present invention will be defined by the appended claims and their equivalents.

Claims

1. A composition for eradicating Helicobacter pylori, the composition comprising zaspirone, a pharmaceutically acceptable salt thereof, a hydrate or solvate thereof, or a mixture thereof, and amoxicillin, a pharmaceutically acceptable salt thereof, or a hydrate thereof.

2. A composition for eradicating Helicobacter pylori, the composition comprising zaspirone, a pharmaceutically acceptable salt thereof, a hydrate or solvate thereof, or a mixture thereof, and clarithromycin or a pharmaceutically acceptable salt thereof.

3. A composition for eradicating Helicobacter pylori, the composition comprising zaspirone, a pharmaceutically acceptable salt thereof, a hydrate or solvate thereof, or a mixture thereof, amoxicillin, a pharmaceutically acceptable salt thereof, or a hydrate thereof, and clarithromycin or a pharmaceutically acceptable salt thereof.

4. The composition for eradicating Helicobacter pylori according to any one of claims 1 to 3, wherein the pharmaceutically acceptable salt of zaspirone is zaspirone citrate.

5. The composition for eradicating Helicobacter pylori according to any one of claims 1 to 3, wherein the composition for eradicating Helicobacter pylori comprises zaspirone citrate in an amount of 5 to 40 mg.

6. The composition for eradicating Helicobacter pylori according to any one of claims 1 and 3, wherein the composition for eradicating Helicobacter pylori comprises amoxicillin, a pharmaceutically acceptable salt thereof, or a hydrate thereof in an amount of 100 to 1000 mg.

7. The composition for eradicating Helicobacter pylori according to any one of claims 2 and 3, wherein the composition for eradicating Helicobacter pylori comprises clarithromycin or a pharmaceutically acceptable salt thereof in an amount of 100 to 1000 mg.

8. The composition for eradicating Helicobacter pylori according to any one of claims 1 to 3, wherein the composition for eradicating Helicobacter pylori is administered one to three times a day.

9. The composition for eradicating Helicobacter pylori according to any one of claims 1 to 3, wherein the composition for eradicating Helicobacter pylori is formulated as a solid oral preparation.

10. The composition for eradicating Helicobacter pylori according to claim 9, wherein the solid oral preparation is any one of tablets, film-coated tablets, capsules, powders, granules, pills, lozenges, oral gels, and oral dissolving films.

11. A composition for treating Helicobacter pylori infection, the composition comprising zaspirone, a pharmaceutically acceptable salt thereof, a hydrate or solvate thereof, or a mixture thereof, and amoxicillin, a pharmaceutically acceptable salt thereof, or a hydrate thereof.

12. A composition for treating Helicobacter pylori infection, the composition comprising zaspirone, a pharmaceutically acceptable salt thereof, a hydrate or solvate thereof, or a mixture thereof, and clarithromycin or a pharmaceutically acceptable salt thereof.

13. A composition for treating Helicobacter pylori infection, the composition comprising zaspirone, a pharmaceutically acceptable salt thereof, a hydrate or solvate thereof, or a mixture thereof, amoxicillin, a pharmaceutically acceptable salt thereof, or a hydrate thereof, and clarithromycin or a pharmaceutically acceptable salt thereof.

14. A method for eradicating Helicobacter pylori, the method comprising the step of administering to a subject in need thereof a pharmaceutically effective amount of a composition comprising a therapeutically effective amount of zaspirone, a pharmaceutically acceptable salt thereof, a hydrate or solvate thereof, or a mixture thereof, and clarithromycin or a pharmaceutically acceptable salt thereof.

15. A method for eradicating Helicobacter pylori, the method comprising the step of administering to a subject in need thereof a pharmaceutically effective amount of a composition comprising a therapeutically effective amount of zaspirone, a pharmaceutically acceptable salt thereof, a hydrate or solvate thereof, or a mixture thereof, amoxicillin, a pharmaceutically acceptable salt thereof, or a hydrate thereof, and clarithromycin or a pharmaceutically acceptable salt thereof.

16. Use of a pharmaceutical composition for eradicating Helicobacter pylori, the pharmaceutical composition comprising zaspirone, a pharmaceutically acceptable salt thereof, a hydrate or solvate thereof, or a mixture thereof, and clarithromycin or a pharmaceutically acceptable salt thereof.

17. Use of a pharmaceutical composition for eradicating Helicobacter pylori, the pharmaceutical composition comprising zaspirone, a pharmaceutically acceptable salt thereof, a hydrate or solvate thereof, or a mixture thereof, amoxicillin, a pharmaceutically acceptable salt thereof, or a hydrate thereof, and clarithromycin or a pharmaceutically acceptable salt thereof.

18. Use of a composition for preparing a medicament for eradicating Helicobacter pylori, the composition comprising zaspirone, a pharmaceutically acceptable salt thereof, a hydrate or solvate thereof, or a mixture thereof, and clarithromycin or a pharmaceutically acceptable salt thereof.

19. Use of a composition for preparing a medicament for eradicating Helicobacter pylori, the composition comprising zaspirone, a pharmaceutically acceptable salt thereof, a hydrate or solvate thereof, or a mixture thereof, amoxicillin, a pharmaceutically acceptable salt thereof, or a hydrate thereof, and clarithromycin or a pharmaceutically acceptable salt thereof.

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