CN1958568A - A kind of compound for preventing or curing infection of helicobacter pylori, preparation method, and application - Google Patents

Abstract

The present invention relates to a compound for treating or preventing diseases or symptoms caused or accompanied by bacterial infection, especially Helicobacter pylori infection, its preparation method and medical application.

Description

A kind of compound for preventing or treating Helicobacter pylori infection, its preparation method and application

technical field

The present invention relates to a compound for preventing or treating diseases or symptoms caused or accompanied by certain bacteria (especially Helicobacter pylori) infection, its preparation method and medical application.

Background technique

Helicobacter pylori (H. pylori) is a slightly anaerobic Gram-negative bacterium that grows in the gastric mucosa. It is reported that about half of the world's population will be infected with this pathogen in their lifetime. In 1983, Marshall and Warren first isolated Helicobacter pylori from gastric mucosal specimens of patients with gastritis. This important discovery changed people's understanding of the etiology of peptic ulcer and greatly promoted the development of gastroenterology research. Therefore won the 2005 Nobel Prize in Physiology or Medicine. It is now clear that Helicobacter pylori is not only the culprit of peptic ulcer, but also the main cause of gastritis and gastric cancer. Therefore, the World Health Organization classifies it as a class I carcinogen.

The mode of Helicobacter pylori infection is closely related to bad living habits, diet and living conditions, and it is very easy to transmit infection to each other in daily life. Although the infection rate of Helicobacter pylori has begun to decline in economically developed countries such as the United States and Western Europe, the number is still on the rise in developing countries (including China). Helicobacter pylori infection can not only cause a series of upper gastrointestinal diseases including chronic gastritis, peptic ulcer, non-ulcer dyspepsia, gastric adenocarcinoma, gastric non-Hodgkin's lymphoma and mucosa-associated lymphoid tissue lymphoma, but also It is associated with the onset or progression of cardiovascular and cerebrovascular diseases, autoimmune diseases and certain skin diseases, thus seriously endangering human health and life.

A single antibiotic is ineffective in treating H. pylori infection and can easily lead to drug resistance. The treatment plan recommended by the medical profession is the combined use of proton pump inhibitors or ranitidine bismuth citrate plus two or three antibiotics, the so-called "triple therapy" or "quadruple therapy". Currently commonly used anti-H. pylori drugs include metronidazole, clarithromycin, amoxicillin, tetracycline, and furazolidone. Due to the long-term irrational use of antibiotics, the resistance of Helicobacter pylori to the above-mentioned drugs (especially metronidazole and clarithromycin) is increasing, which seriously affects the clinical treatment effect.

The huge number of infected people around the world and the difficulties faced in clinical treatment call for the generation of new drugs against Helicobacter pylori infection. A candidate drug of this type with good development prospects should reflect the following characteristics: (1) a new structure and a mechanism of action different from existing drugs: (2) strong antibacterial activity and acid stability; (3) narrow antibacterial spectrum and high selectivity , it is still effective against drug-resistant Helicobacter pylori; (4) Drug resistance is not easy to occur after repeated use.

Aminopeptidases are a class of proteases that can catalyze the cleavage of N-terminal amino acid residues of polypeptides and proteins. They are widely found in prokaryotic and eukaryotic cells. The transport, localization and functional regulation in cells are extremely important, and they are necessary for normal physiological functions of cells. Therefore, aminopeptidase is a potential drug target. It has been reported that some aminopeptidase inhibitors have the activity of inhibiting the growth of bacteria or parasites. Leucyl aminopeptidase (LAP), cloned from Helicobacter pylori, is a rare regulatory biological enzyme in the aminopeptidase family, and it is speculated that it is essential for maintaining the normal life activities of Helicobacter pylori.

The invention starts from the recombinantly expressed Helicobacter pylori leucine aminopeptidase, establishes a high-throughput screening model based on relevant enzyme kinetics, and discovers a class of inhibitors specifically acting on the enzyme through large-scale random screening. Experiments have shown that these compounds have certain antibacterial activity against Helicobacter pylori. The invention provides a related technology for synthesizing a novel anti-helicobacter pylori drug and its application.

Contents of the invention

The object of the present invention is to provide a compound of formula I or a pharmaceutically acceptable salt thereof capable of inhibiting bacterial infection (especially Helicobacter pylori);

Another object of the present invention is to provide a method for preparing the compound of formula I;

Another object of the present invention is to provide a pharmaceutical composition containing a compound of formula I;

Another object of the present invention is to provide the use of the compound of formula I for the preparation of medicines for preventing or treating diseases such as peptic ulcer.

The present invention provides a series of compounds that prevent the growth of certain bacteria (especially Helicobacter pylori) by inhibiting the activity of leucine aminopeptidase, increasing the members of antibiotics. The present invention relates to a compound having the following molecular formula I, or a pharmaceutically acceptable salt thereof:

Wherein Ar is any of the following substituents:

R ₁ and R ₂ are any one, two or three of the following substituents: including C ₁ -C ₄ alkyl, nitro, cyano, carboxyl, ester, aldehyde, halogen, hydroxyl, C ₁ - _C4 alkoxy, amine, amido, carbonamido, mercapto, methylthio, ethylthio; aryl; 2-, 3-, or 4-position pyridyl; furyl; pyranyl; Thienyl; pyrrolyl; containing C ₁ -C ₄ alkyl, nitro, cyano, carboxyl, ester, aldehyde, halogen, hydroxyl, alkoxy, amine, amido, carbonamido, mercapto , methylthio, ethylthio, any one, two or three substituted aryl groups; containing C ₁ -C ₄ alkyl, nitro, cyano, carboxyl, ester, aldehyde, halogen , hydroxyl, alkoxy, amine, amido, carbonamido, mercapto, methylthio, ethylthio, any one, two or three substituted 2-, 3-, or 4-position pyridine group; containing C ₁ -C ₄ alkyl group, nitro group, cyano group, carboxyl group, ester group, aldehyde group, halogen, hydroxyl group, alkoxy group, amine group, amido group, carbon amido group, mercapto group, methylthio group Any one, two or three substituted furyl groups including C ₁ -C ₄ , nitro, cyano, carboxyl, ester, aldehyde, halogen, hydroxyl, alkane Any one, two or three substituted pyranyl groups including oxy, amine, amido, carbonamido, mercapto, methylthio, ethylthio; containing C ₁ -C ₄ alkyl, Any one, two, or Three substituted thienyl groups; containing C ₁ -C ₄ alkyl, nitro, cyano, carboxyl, ester, aldehyde, halogen, hydroxyl, alkoxy, amine, amido, carbonamido, Any one, two or three substituted pyrrolyl groups including mercapto, methylthio and ethylthio. X is O, NH, NCH ₃ or S; n is an integer of 0-8;

Y is H, O, S, NH, NCH ₃ , or NOH;

Z is amino, C ₁ -C ₄ monoalkyl or dialkyl substituted amino, or includes aryl, 2-, 3-, or 4-position pyridyl, furyl, pyryl, thienyl, pyrrolyl in Any one or two substituted amino groups within.

Preferably, the series of compounds have, but are not limited to, the following molecular formula II:

The compounds contained in the general chemical structure formula of the present invention can be prepared by the following general method (see Journal of medicinal chemistry, 1993, 36 (17): 2485-2493 for specific methods):

Add 1eq ArXH, 1-2eq potassium carbonate to an appropriate amount of acetone, dichloromethane, chloroform or ethyl acetate to reflux, slowly add 1-1.5eq Cl(CH ₂ )nCN in acetone, dichloromethane, chloroform or acetic acid ethyl ester solution. After the dropwise addition, continue to stir and reflux for 2-20 hours to obtain intermediate 2.

Take 1eq of intermediate 2 and dissolve in an appropriate amount of absolute ethanol, boil and reflux. 10-20eq of sodium metal was added slowly. The mixture was poured into dilute hydrochloric acid, and an appropriate amount of absolute ethanol was added. Filter out the inorganic filtrate, add 85% absolute ethanol to cool, precipitate the hydrochloride of the amine, and filter.

Mix intermediate 2 with an appropriate amount of 60-90% sulfuric acid, stir and react at 70-100° C. for more than 2 hours, and filter. Transfer the filter cake to the reaction bottle, add ammonia water to neutralize under stirring, control the neutralization temperature not to exceed 50°C, and maintain the neutralized pH value at about 5-7, filter, and filter out solid ammonium sulfate. On cooling, a solid precipitated out to give the amide.

Dissolve 1eq of intermediate 2 and 2eq of hydroxylamine hydrochloride in an appropriate amount of dimethyl sulfoxide or DMF, add 2-4eq of methanol solution of sodium methoxide under stirring, and heat up to 60-90°C for more than 1 hour. Excess methanol was distilled off under reduced pressure, the residue was stirred with water, and ethyl acetate was added to separate layers, the aqueous layer was washed with ethyl acetate, the organic phases were combined, dried, concentrated, and the product oxime was obtained by column chromatography.

Furthermore, preferably, the series of compounds or their pharmaceutically acceptable salts are provided in the form of a pharmaceutical composition, either alone or in combination with pharmaceutically acceptable carriers or excipients. The present invention also provides a kit comprising the above compounds for preventing or treating diseases or symptoms caused or accompanied by bacterial infections, such as Helicobacter pylori.

In another aspect, the present invention relates to a combined preparation, which includes a compound that selectively inhibits bacterial growth, especially the activity of Helicobacter pylori leucine aminopeptidase, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient. Preferably, the combination preparation comprises a compound or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient.

The present invention also provides a kit comprising the above-mentioned combination preparation. The present invention further provides methods and kits for treating or preventing diseases or symptoms caused or accompanied by bacterial infection, such as Helicobacter pylori infection, using the above-mentioned combined preparation.

Detailed ways

In order to clarify the content of the invention and not to limit it, the invention is divided into the following subsections for a detailed description.

A definition

Unless otherwise defined, the technical and scientific terms used in the present invention have the same meaning as commonly understood in the field to which the present invention belongs. All patents, applications, published applications and other publications and sequences from GenBank and other databases mentioned herein are incorporated by reference in their entirety. If the definitions set forth in this section are contrary to, or inconsistent with, the definitions set forth in all patents, applications, published applications, and other publications and sequences incorporated and referenced in GenBank and other databases referenced by this patent, the definitions set forth in this section shall The stated definition shall prevail.

As used in the present invention, "a" or "an" means "at least one" or "one or more".

As used in the present invention, "Helicobacter pylori" refers to a kind of microanaerobic Gram-negative bacillus strains that grow in the gastric mucosa.

As used in the present invention, "diseases or symptoms caused by or accompanied by Helicobacter pylori infection" refers to diseases and symptoms caused by Helicobacter pylori infection alone, or jointly caused by other genetic and/or acquired factors and conditions.

The "effective amount" of a compound used in the present invention for treating a specific disease refers to the amount sufficient to improve or alleviate the symptoms associated with the disease to some extent. This dose can be administered as a single dose or as a regimen. This dose is curative of the disease, but is typically administered to ameliorate the symptoms. Repeat dosing may be necessary to improve symptoms.

As used in the present invention, "pharmaceutically acceptable salts, esters or other derivatives" includes any salts, esters or derivatives that can be easily prepared by those skilled in the field of medicine and chemical engineering by known methods. The compounds thus derived and produced can be administered to animals and humans without toxic effects. The compound is either pharmaceutically active or a prodrug.

As used herein, "treating" refers to amelioration of disease and symptoms in any way, or other beneficial changes. Treatment also includes the use of the compounds of the invention in medicine.

As used herein, "improving" the symptoms of a particular disease by administering a particular pharmaceutical composition means that any relief, whether permanent, temporary, long-term, or transient, is attributable to or associated with the pharmaceutical composition. related to the application.

As used in the present invention, "substantially pure" means that it is uniform enough that no impurities can be detected by the standard analytical methods used by those skilled in the field of medicine and chemical industry to evaluate the purity, such as thin layer chromatography (TLC), gel Electrophoresis and high performance liquid chromatography (HPLC). Or sufficiently pure means that even further purification does not change the detectable physicochemical properties of the substance, such as enzymatic activity and biological activity. Methods for purifying compounds to obtain substantially chemical purity are well known to those skilled in the pharmaceutical and chemical arts. A substantially chemically pure compound may, however, be a mixture of stereoisomers or isomers. In such cases, further purification may increase the specific activity of the compound.

As used herein, "prodrug" refers to an in vivo administered compound that can be metabolized, or converted into a biologically, pharmaceutically or therapeutically active form. To make prodrugs, the pharmaceutically active compound is modified such that the active compound is reproduced by metabolic processes. Prodrugs can be engineered to alter their metabolic stability, or their transport properties, to mask their side effects or toxicity, to modify the taste of the drug, or to alter other properties. With the knowledge of pharmacokinetics and drug metabolism in vivo, once the pharmacologically active compound is known, those skilled in the field of pharmaceutical and chemical engineering can design the prodrug of the compound. [See Nogrady, Medicinal Chemistry A Biochemical Approach, Oxford University Press, New York, pages 388-392 (1985)].

The term "substantially" is the same or uniform or similar. According to the understanding of the relevant technology by those skilled in the field of pharmaceutical and chemical engineering, the context can be changed, and generally at least 70%, preferably at least 80%, more preferably at least 90%, Optimally at least 95% identical.

As used herein "composition" means any mixture. Can be solution, suspension, liquid, powder, ointment, aqueous, non-aqueous or any combination thereof.

As used herein, "combination" refers to any combination of two or more.

The term "subject" as used herein includes humans and animals such as dogs, cats, cows, pigs, rodents and the like. Experienced practitioners will understand that subjects are suitable and willing to treat and prevent diseases or symptoms caused by or accompanied by bacterial infection, such as Helicobacter pylori infection.

Any abbreviations for protective groups, amino acids and other compounds used herein are consistent with their commonly used, recognized abbreviations or biochemical names issued by the IUPAC-IUB committee, unless otherwise specified. [See Biochem. 11:1726 (1972)].

B antibacterial agent

The present invention adds to the series of antibiotics by providing drugs that have an inhibitory effect on the activity of certain bacterial leucine aminopeptidases. One aspect of the present invention relates to a compound having the following molecular formula I, or a pharmaceutically acceptable salt thereof:

Wherein Ar is any of the following substituents:

R ₁ and R ₂ are any one, two or three of the following substituents: including C ₁ -C ₄ alkyl, nitro, cyano, carboxyl, ester, aldehyde, halogen, hydroxyl, C ₁ - _C4 alkoxy, amine, amido, carbonamido, mercapto, methylthio, ethylthio; aryl; 2-, 3-, or 4-position pyridyl; furyl; pyranyl; Thienyl; pyrrolyl; containing C ₁ -C ₄ alkyl, nitro, cyano, carboxyl, ester, aldehyde, halogen, hydroxyl, alkoxy, amine, amido, carbonamido, mercapto , methylthio, ethylthio, any one, two or three substituted aryl groups; containing C ₁ -C ₄ alkyl, nitro, cyano, carboxyl, ester, aldehyde, halogen , hydroxyl, alkoxy, amine, amido, carbonamido, mercapto, methylthio, ethylthio, any one, two or three substituted 2-, 3-, or 4-position pyridine group; containing C ₁ -C ₄ alkyl group, nitro group, cyano group, carboxyl group, ester group, aldehyde group, halogen, hydroxyl group, alkoxy group, amine group, amido group, carbon amido group, mercapto group, methylthio group Any one, two or three substituted furyl groups including C ₁ -C ₄ , nitro, cyano, carboxyl, ester, aldehyde, halogen, hydroxyl, alkane Any one, two or three substituted pyranyl groups including oxy, amine, amido, carbonamido, mercapto, methylthio, ethylthio; containing C ₁ -C ₄ alkyl, Any one, two, or Three substituted thienyl groups; containing C ₁ -C ₄ alkyl, nitro, cyano, carboxyl, ester, aldehyde, halogen, hydroxyl, alkoxy, amine, amido, carbonamido, Any one, two or three substituted pyrrolyl groups including mercapto, methylthio and ethylthio. X is O, NH, NCH ₃ or S; n is an integer of 0-8:

Y is H, O, S, NH, NCH ₃ , or NOH;

Z is amino, C ₁ -C ₄ monoalkyl or dialkyl substituted amino, or includes aryl, 2-, 3-, or 4-position pyridyl, furyl, pyryl, thienyl, pyrrolyl in Any one or two substituted amino groups within.

Preferably, the series of compounds have, but are not limited to, the following molecular formula II:

The compounds of the present invention may be in a particular stereoisomer, eg cis or trans configuration, or mixtures thereof. Compounds contemplated here include all classes of pharmaceutically active compounds, or solutions or mixtures thereof. Also included are their hydration types, such as aqueous solutions of these compounds, hydrolysates or ionized products; and these compounds may contain varying numbers of bound water molecules.

The compounds of the invention may be prepared or synthesized according to any suitable method. Preferably, the compounds are prepared using the synthetic methods cited in Section E below.

Also, preferably, the compound or a pharmaceutically acceptable salt thereof is provided in the form of a pharmaceutical composition, either alone or in combination with a pharmaceutically acceptable carrier or excipient.

The compounds of the present invention may be prepared with any suitable acid in the form of a pharmaceutically acceptable salt thereof. For example; inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, phosphoric acid, etc. can be used. Among other examples, organic acids such as formic acid, acetic acid, propionic acid, benzoic acid, maleic acid, fumaric acid, succinic acid, tartaric acid, citric acid and the like can be used. As another example, alkylsulfonic acids such as methanesulfonic acid, ethylsulfonic acid and the like can be used. As another example, arylsulfonic acids such as benzenesulfonic acid, p-toluenesulfonic acid, etc. can be used.

Any subject may be treated with the method, preferably a mammal, more preferably a human.

The method can be used to prevent and treat any diseases and symptoms caused by Helicobacter pylori infection. Preferred diseases are chronic gastritis, peptic ulcer, non-ulcer dyspepsia, gastric adenocarcinoma, gastric non-Hodgkin's lymphoma, and mucosa-associated lymphoid tissue lymphoma, among others.

When preventing or treating diseases and symptoms caused by the above-mentioned bacterial infections, the compound of the present invention can be used alone or in combination with other proton pump inhibitors, bismuth agents and antibiotics that have been or will be on the market. Any suitable proton pump inhibitors, bismuth agents and antibiotics may be used in combination with the compounds of the present invention. Typical antibiotics are metronidazole, clarithromycin, amoxicillin, tetracycline, and furazolidone; other antibiotics include penicillins and their derivatives, cephalosporins and their derivatives, macrolide antibiotics, aminoglycoside antibiotics, tetracyclines Class antibiotics, new glycopeptide antibiotics, polypeptide antibiotics, new quinolone antibiotics, oxazolidinone antibiotics, dihydrofolate reductase inhibitors and various antibiotic enhancers, etc.

In a preferred embodiment of the invention, the above-mentioned antibiotics are not administered when using the compounds of the invention. More preferably, the compounds of the present invention are used to treat or prevent diseases and symptoms caused by resistant strains induced by the above-mentioned antibiotics.

The compounds of the invention may be administered by any suitable method alone, or in combination with other suitable antibiotics. For example, the compound of the present invention, or a pharmaceutically acceptable salt thereof can be administered by intracavitary injection, subcutaneous injection, intravenous injection, intramuscular injection, intradermal injection, orally or topically.

In specific embodiments, the method further comprises diagnosing and assessing the prognosis of the bacterial infection in the subject. Any suitable method may be used for diagnosing and assessing the prognosis of bacterial infection. Diagnosis and prognosis can be based on detection and/or identification of any or all bacterial proteins, such as its enzymes, antigens, antibodies, nucleic acids or other pathological and clinical markers and symptoms. For example, the diagnostic or prognostic methods disclosed in International Patent WO 01/44815 and US Patent 5,571,674 can be used.

C Combination Formulations, Kits and Methods of Combination

On the other hand, the present invention also relates to a combination preparation, which comprises a compound, or a pharmaceutically acceptable salt thereof, which selectively inhibits the activity of leucine aminopeptidase of bacteria, especially Helicobacter pylori, and one or more antibiotics.

Preferably, the combination includes a compound or a pharmaceutically acceptable salt thereof and one or more antibiotics, the compound having the above-mentioned formulas I, II and the defined substituents.

Any suitable antibiotic may be used in the combination formulations of the invention. In a particular embodiment, one or more of the above-mentioned antibiotics may be included in the combination formulation of the present invention.

In another specific embodiment, there is provided a method of treating or preventing a disease or symptom caused or accompanied by bacterial infection, such as Helicobacter pylori infection, the method comprising administering an effective Amount of the above combination preparation, or a pharmaceutically acceptable salt thereof, so as to treat or prevent the above diseases or symptoms.

In another specific embodiment, there is provided a kit comprising a compound of the present invention or a pharmaceutically acceptable salt thereof and the use of said compound or a pharmaceutically acceptable salt thereof for the prevention and treatment of bacterial infections, such as Helicobacter pylori Instructions for use for bacterial infections, diseases or symptoms caused.

In yet another embodiment, a kit is provided, comprising the above combination preparation and instructions for using the combination preparation to treat or prevent diseases or symptoms caused by bacterial infections, such as Helicobacter pylori infection.

D formulation and dosage

According to the present invention, the compounds of the present invention, alone or in combination with other agents, carriers or excipients, are formulated for any suitable route of administration, such as intracavitary injection, subcutaneous injection, intravenous injection, intramuscular injection, intradermal injection Injected, taken orally, or topically. The method may employ formulations for parenteral administration, administered as single doses in ampoules, or in multi-dose containers by injection with added buffer. The formulations may take such forms as suspensions, solutions, or emulsions in oily or aqueous vehicles. The formulation may contain formulatory agents such as suspending, stabilizing and/or dispersing agents. In addition, before use, the active ingredient can be formulated in powder form with a suitable carrier, sterile pyrogen-free water or other solvents. Foams, gels, ointments, salves, transdermal patches, or creams may be used for topical administration of the present invention.

The size of the dose for treatment or prophylaxis will vary depending on the severity of the condition and the route of administration. Dosage and frequency of administration will vary according to age, weight, health status and individual response of the patient.

It should be pointed out (and the treating physician should also know), that according to the toxicity and side effects, necessary measures must be taken to terminate, interrupt or reduce the treatment dose. On the contrary, if the clinical response is not obvious (excluding toxicity and side effects), doctors should adjust the treatment plan appropriately and increase the dose.

Any suitable route of administration can be used. Dosage forms include tablets, lozenges, bean capsules, dispersions, suspensions, solutions, capsules, films and the like.

In practice, the compounds of the present invention, alone or in combination with other preparations, can be mixed with pharmaceutical carriers or excipients, such as β-cyclodextrin and 2-hydroxy-propyl-β-cyclodextrin, in accordance with general pharmaceutical mixing techniques. Finely blend. According to the needs of administration, general carriers, special carriers for topical or parenteral routes can be used. Preparation of parenteral dosage forms, such as compositions for intravenous injection or infusion, can use similar pharmaceutical media, water, glycol, oil, buffer, sugar, preservatives, liposomes, etc. known to those skilled in the art . Examples of such parenteral compositions include, but are not limited to, 5% w/v dextrose, saline or other solutions. The total dose of the compound of the present invention, administered alone or in combination with other formulations, may be administered via intravenous injection vials with a volume ranging from approximately 1 ml to 2000 ml. The amount of diluent will vary depending on the total dose administered.

The invention also provides a kit for implementing a treatment regimen. The kit contains an effective dose of a compound of the invention, alone or in combination with other agents, in a pharmaceutically acceptable form, contained in one or more containers. The preferred pharmaceutical form is in combination with sterile saline, dextrose solution, buffered solution, or other pharmaceutically acceptable sterile liquid. Alternatively, the composition may be lyophilized or dried; in this case, the kit optionally further comprises a pharmaceutically acceptable solution, preferably sterile, in a container to reconstitute the complex Solutions are formed for injection purposes. Typical pharmaceutically acceptable solutions are saline solution and dextrose solution.

In another embodiment, the kit according to the present invention further comprises a needle or syringe packaged, preferably in sterile form, and/or a packaged alcohol pad for the injectable composition. Instructions for use by a physician or patient can optionally be included.

E embodiment

Experimental Instruments and Reagents

HP1100 HPLC system with binary gradient pump, online vacuum degasser, autosampler, column oven and photodiode array detector. The chromatographic column is ZORBAX ODS (4.6×250mm), the mobile phase is methanol/water (80:20), the flow rate is 1ml/min, and the detection wavelength is 254nm.

All solvents were HPLC grade. The MS spectrum was measured by a PE API 2000LC/MS/MS mass spectrometer. ¹ H-NMR data were measured by the Analysis and Testing Center of Shanghai Institute of Materia Medica, Chinese Academy of Sciences. All synthetic starting materials were commercially available

Example 1

0.020 mol of compound 1 and 0.025 mol of potassium carbonate were added to 50 ml of acetone for reflux, and 10 ml of 0.026 mol of Cl(CH ₂ )nCN in acetone was slowly added dropwise. After the dropwise addition was completed in 30 minutes, the mixture was stirred and refluxed for 18 hours, cooled and concentrated. A layer of 50ml ethyl acetate and 50ml water was added. The aqueous layer was washed three times with ethyl acetate (20 ml), the organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and column chromatographed to obtain a pale yellow solid. The following compounds were synthesized by this method:

¹ HNMR (CDCl ₃ ): 3.66(s, 2H), 7.54(m, 3H), 7.81(m, 3H), 8.05(s, 1H).

¹ HNMR (CDCl ₃ ): 4.68 (s, 2H), 7.08 (m, 2H), 7.35 (m, 2H), 7.68 (m, 3H)

Dissolve 2.64mmol of compound 2 and 5.29mmol of hydroxylamine hydrochloride in an appropriate amount of dimethyl sulfoxide, add 4.1ml of methanolic sodium methoxide solution (1.0g of metallic sodium dissolved in 20ml of methanol) with stirring, and heat up to 80°C for 1.5 hours . Methanol was distilled off under reduced pressure, the residue was added with 50ml of water and stirred for 1 hour, 50ml of ethyl acetate was added to separate layers, the aqueous layer was washed 3 times with ethyl acetate (20ml), the organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the column layer was An off-white solid was precipitated. The following compounds were synthesized by this method:

¹ HNMR (CDCl ₃ ): 3.68(s, 2H), 4.90(s, 2H), 7.45(s, 3H), 7.76(s, 3H), 7.85(s, 1H).

¹ H NMR (DMSO-D ₆ ): 4.47 (s, 2H), 5.62 (s, 2H), 7.17 (m, 1H), 7.38 (m, 3H), 7.77 (m, 3H)

The activity of embodiment 2 novel antibacterial series compounds

Helicobacter pylori leucine aminopeptidase is an allosteric enzyme that can specifically hydrolyze peptide bonds with amino acid residues such as Leu-, and its activity is regulated by divalent cations. Under the reaction conditions of 37°C, 50mM Tris-HCl (pH 8.0) buffer and 0.5mM MnCl2, leucine aminopeptidase can catalyze the hydrolysis of the substrate L-leu-p-nitroanilide, and the generated product pNA (p-nitro Aniline) has characteristic light absorption at 405 nm. The absorbance was detected by Spectra MAX 340 Microplate Reader, and the relative inhibition rate of the test sample to leucine aminopeptidase was converted [Lei Dong, Ni Cheng, Ming-Wei Wang, Junfeng Zhang, Chang Shu and De-Xu Zhu. The leucyl aminopeptidase from Helicobacter pylori isan allosteric enzyme. Microbiology, 2005, 151: 2017-2023]. Accordingly, through biological activity screening and re-screening verification of 32,000 samples, a series of compounds with inhibitory activity on leucine aminopeptidase were found, and their half inhibitory concentration (IC50) was between 2-9 μM.

In order to study the influence of the above compounds on the life cycle of Helicobacter pylori, the minimum inhibitory concentration (MIC) of them to the standard strain of Helicobacter pylori ATCC43504 was determined by agar dilution method. The results showed that the compound NC00072450 had certain anti-Helicobacter pylori activity, and its MIC value was 32 μg/ml (see the table below).

Table 1. Summary of the inhibitory effect of related compounds of the present invention on leucine aminopeptidase and Helicobacter pylori standard strain ATCC43504

(Bestatin is a positive control)

The above examples are for illustrative purposes only, and the scope of the present invention is not limited thereto. Modifications will be obvious to those skilled in the fields of pharmaceutical and chemical engineering, and the present invention is limited only by the scope of the appended claims.

Claims (7)

1. acceptable salt on a compound or its pharmacology, this compound has following molecular formula I:

Wherein Ar is following any one substituting group:

R ₁, R ₂For following arbitrarily any one, two or three substituting groups: comprise C ₁-C ₄Alkyl, nitro, cyano group, carboxyl, ester group, aldehyde radical, halogen, hydroxyl, C ₁-C ₄Alkoxyl group, amido, amide group, carbonamido, sulfydryl, methylthio group, ethylmercapto group; Aryl; 2-, 3-or 4-position pyridyl; Furyl; Pyranyl; Thienyl; Pyrryl; Contain and comprise C ₁-C ₄Alkyl, nitro, cyano group, carboxyl, ester group, aldehyde radical, halogen, hydroxyl, alkoxyl group, amido, amide group, carbonamido, sulfydryl, methylthio group, ethylmercapto group at interior any one, the aryl of two or three replacements; Contain and comprise C ₁-C ₄Alkyl, nitro, cyano group, carboxyl, ester group, aldehyde radical, halogen, hydroxyl, alkoxyl group, amido, amide group, carbonamido, sulfydryl, methylthio group, ethylmercapto group at 2-, 3-or the 4-position pyridyl of interior any one, two or three replacements; Contain and comprise C ₁-C ₄Alkyl, nitro, cyano group, carboxyl, ester group, aldehyde radical, halogen, hydroxyl, alkoxyl group, amido, amide group, carbonamido, sulfydryl, methylthio group, ethylmercapto group at interior any one, the furyl of two or three replacements; Contain and comprise C ₁-C ₄Alkyl, nitro, cyano group, carboxyl, ester group, aldehyde radical, halogen, hydroxyl, alkoxyl group, amido, amide group, carbonamido, sulfydryl, methylthio group, ethylmercapto group at interior any one, the pyranyl of two or three replacements; Contain and comprise C ₁-C ₄Alkyl, nitro, cyano group, carboxyl, ester group, aldehyde radical, halogen, hydroxyl, alkoxyl group, amido, amide group, carbonamido, sulfydryl, methylthio group, ethylmercapto group at interior any one, the thienyl of two or three replacements; Contain and comprise C ₁-C ₄Alkyl, nitro, cyano group, carboxyl, ester group, aldehyde radical, halogen, hydroxyl, alkoxyl group, amido, amide group, carbonamido, sulfydryl, methylthio group, ethylmercapto group at interior any one, the pyrryl of two or three replacements.X is O, NH, NCH ₃Or S;

N is the integer of 0-8;

Y is H, O, S, NH, NCH ₃, or NOH;

Z is amino, C ₁-C ₄Monoalkyl or two alkyl-substituted amino, or comprise aryl, 2-, 3-or 4-position pyridyl, furyl, pyranyl, thienyl, pyrryl is at interior any one or the amino of two replacements.

2. compound as claimed in claim 1 is characterized in that having, but is not limited to following molecular formula II:

3. the preparation method of compound as claimed in claim 1 and pharmacy acceptable salt thereof comprises the following steps:

4. one kind contains the effective dose of claim 1 or 2 described compounds and pharmacy acceptable salt thereof and the composition that assistant agent, vehicle are formed.

5. composition according to claim 4 is characterized in that comprising proton pump inhibitor, bismuth agent and microbiotic.Typical microbiotic refers to metronidazole, clarithromycin, amoxycilline Trihydrate bp, tsiklomitsin and Nifurazolidone; Other microbiotic comprise penicillin and derivative, cynnematin and derivative thereof, macrolide antibiotics, aminoglycoside antibiotics, tetracycline antibiotics, new glycopeptide antibiotics, polypeptide antibiotics, Novel Quinolone class microbiotic, oxazolidine ketone microbiotic, dihydrofolate reductase inhibitor and various microbiotic toughener.

6. the purposes of compound as claimed in claim 1 and pharmacy acceptable salt thereof is used in the medicine of preparation prevention and treatment bacterial infection disease.

7. purposes according to claim 5, it is characterized in that being used for preparing by bacterial infection disease is to be used by the disease medicament that helicobacter pylori infection causes.