CN109503569B - Thiazole derivative and preparation method and application thereof - Google Patents

Abstract

The invention provides thiazole derivatives, a preparation method and application thereof, wherein the compounds have a structure shown in a formula (V):

wherein A is a cyclic or chain group containing a nitrogen atom selected from

And

wherein one side containing a nitrogen atom is linked to X; x is S or C; y is O or S; n is 1 or 2; wherein, when A is a cyclic group containing 1 nitrogen atom, X is not C; r is R¹or-NHR²；R¹、R²Each independently selected from C_1～8Straight or branched alkyl, C_2‑8Straight-chain or branched alkenyl, C_2‑8Straight-chain or branched alkynyl, heteroalkyl, C_3‑12Cycloalkyl, aryl, heteroaryl; said cycloalkyl, aryl, heteroaryl being unsubstituted or selected from halogen, nitro, amino, C_1～8Straight or branched alkyl, C_2‑8Straight-chain or branched alkenyl, C_2‑8One or more of straight chain or branched chain alkynyl, heteroalkyl and alkoxy.

Description

Thiazole derivative and preparation method and application thereof

Technical Field

The invention relates to the field of medicines, and particularly relates to a thiazole derivative, and a preparation method and application thereof.

Background

Oral health is one of ten major signs of human health, and caries (Dental caries) is one of common oral bacterial infectious diseases which seriously affect human health, and has the characteristics of high morbidity, wide distribution, low treatment rate and the like. Statistically, more than 90% of all people worldwide will experience at least one caries insult during their lifetime. Caries is easy to ignore because it is not an infectious and lethal disease, but the chronic development of caries will cause irreversible damage to hard tooth tissues, which not only easily causes oral diseases such as periapical periodontitis, pharyngitis, periodontal disease, pulpitis, cellulitis and the like, and causes complications such as alveolar bone infection, bacteremia and the like, but also has close relationship with systemic diseases such as cerebral apoplexy, heart disease, diabetes, digestive system diseases and the like. Not only pain for individuals but also enormous medical burden for society (Selwitz RH, et al lancet,2007, 51).

In China, with the improvement of the living standard of people, the dietary structure is changed, the intake of high-sugar foods and beverages is continuously increased, the incidence of caries is greatly increased at each age stage, and the prevention and treatment situation is very severe. According to the fourth national oral health epidemiological investigation result published by 9-month-Wei-Ji-Commission in 2017, the caries rate of children of 5 years old reaches 70.9%, the caries rate of teenagers of 12 years old reaches 66%, the caries rate of middle-aged and young reaches 88.1%, and the caries rate of old people reaches 98.4%. The World Health Organization (WHO) has put cardiovascular diseases, cancers and caries into three major categories of non-infectious diseases which need to be mainly prevented and treated, and the prevention and treatment of caries is marked to be a health problem which is commonly concerned globally.

Caries is caused by the presence of a biofilm containing cariogenic bacteria, such as plaque, a complex biofilm. Cariogenic bacteria living therein break down fermentable carbohydrates (primarily sucrose) into organic acids, resulting in progressive mineralization and disintegration destruction of tooth tissue. Of the several major cariogenic bacteria (Streptococcus mutans, Actinomyces, Lactobacillus acidophilus) that have been identified, Streptococcus mutans (Streptococcus mutans for short) has been identified as the major cariogenic pathogen. The mutan has acid production and acid resistance, can synthesize extracellular polysaccharide by sucrose under the action of glucosyltransferase (Gtfs), and can be adhered to the surface of a tooth coated by glucan, so that the mutan becomes three most important cariogenic virulence factors (acid production, acid resistance and synthesis of extracellular polysaccharide) involved in cariogenic pathogenesis. Compared with the planktonic (planktonic) growth mode, most bacteria tend to grow in a biofilm (biofilm) mode in the natural environment. Plaque biofilm can be viewed as a micro-ecological environment that adheres to the tooth surface, in which various normal and cariogenic bacteria grow, and is also a recognized initiating factor in the development of caries. Studies have demonstrated that individuals with plaque biofilms containing low levels of mutans are more resistant to exogenous colonization by other cariogenic pathogens, and therefore studies of the effects of mutans biofilms are of particular importance in the prevention and treatment of caries.

The research and development of novel anti-caries compounds which can inhibit oral cariogenic streptococci, inhibit biofilm related to cariogenic and specific cariogenic virulence factors thereof and have no influence on oral normal bacteria are important research directions in the field of caries prevention and treatment.

Disclosure of Invention

Aiming at the defects of the prior art, the invention aims to provide a novel thiazole compound, a preparation method thereof, an effect of resisting oral cariogenic bacteria and inhibiting biofilm related to cariogenic and a specific cariogenic virulence factor, and provides theoretical and experimental basis for developing a medicament for preventing and treating clinical caries.

The invention is proved by in vitro activity detection: the compound of the invention can better inhibit the activity of oral cariogenic bacteria, inhibit the formation of a variable chain bacteria biofilm, obviously reduce the growth activity of a mature variable chain bacteria biofilm, effectively inhibit the acid production of the variable chain bacteria, has no influence on normal oral bacteria, and is an anti-cariogenic medicament with potential development prospect.

Specifically, the technical scheme of the invention is as follows:

in a first aspect of the invention, the invention provides a compound having the structure shown in formula (V):

wherein A is a group containing 1 nitrogen atom; the nitrogen-containing group can be in a ring or chain structure;

x is S or C; y is O or S; n is 1 or 2;

for example, when X is S, n ═ 2 and Y is O, -X (═ Y)_nIs namely-SO₂Structure; when X is C, -X (═ Y)_nIs of the structure-C ═ O or-C ═ S.

R is R¹or-NHR²；

R¹、R²Each independently selected from C_1-8Straight or branched alkyl, C_2-8Straight-chain or branched alkenyl, C_2-8Straight-chain or branched alkynyl, heteroalkyl, C_3-12Cycloalkyl, aryl, heteroaryl;

said cycloalkyl, aryl, heteroaryl being unsubstituted or selected from halogen, nitro, amino, C_1-8Straight or branched alkyl, C_2-8Straight-chain or branched alkenyl, C_2-8One or more of straight chain or branched chain alkynyl, heteroalkyl and alkoxy.

In the compound of formula (V), A is selected from

Wherein m is a positive integer from 1 to 6, for example m can be 1, 2,3, 4, 5 or 6. Wherein

In (2), one side containing a nitrogen atom is connected with X; and when A is

When X is not C.

The applicant is in the studyIt is found that when A is

And when X is C or one side containing nitrogen atoms is connected with carbonyl, the compounds cannot generate the selectivity to cariogenic bacteria expected by the invention, and the influence on normal oral bacteria is large. For example, when the structure is

(XQH-3-5) or

(XQH-3-14), the effect of killing cariogenic bacteria is not good, the bacteriostatic activity to a plurality of cariogenic bacteria is not good, the bacteriostatic effect is shown in the invention table 1, under the connection relation, XQH-3-14 without substitution on the benzene ring can obtain a little better bacteriostatic activity than XQH-3-5 with substitution of halogen, compared with the effect of killing S.mutans UA159, the effect of XQH-3-14 is a little better, but the MBC is only 64 mug/mL, still not ideal. For example, when A is

And the nitrogen end of the compound is far away from the X side, when the compound is connected with the carbonyl of the amide, the MIC of the compound on S.mutans UA159 is generally more than 32 mug/mL, the MIC of the compounds in various connection modes is 64 mug/mL, and the MBC of the compound in the connection modes is generally more than 16 mug/mL.

In addition, it should be noted that the immobilization groups present in the general structures of the present invention are all indispensable parts of the core structure of the present invention, and they exert the pharmaceutical activity as a whole by combining the linking means and the group substitution means described in the present invention. For example, the nitro group on the thiazole ring, which in the structure of the present invention is involved in the inhibition of the formation of the streptococcus mutans biofilm by the present invention, for example, when it is removed or transferred to another group, such as a simple transfer to an R group, the bacteriostatic and bactericidal effects of the present invention and the inhibitory and clearing effects on the formation of the streptococcus mutans biofilm are not achieved.

Aryl in the context of the present invention refers to an aromatic carbocyclic group, further selected from aromatic rings containing 6 to 8 carbon atoms, further a benzene ring.

The heteroaryl group in the present invention refers to an aromatic heterocyclic ring, which may be monocyclic or polycyclic, and further, the heteroaryl group includes: thienyl, pyrrolyl, furyl, pyridyl, pyrazine, thiazolyl, quinolinyl, isoquinoline, pyrimidinyl, tetrazolyl, benzofuryl, benzothiazolyl, indolyl, piperazinyl, tetrahydropiperazinyl, and the like.

The alkyl group in the present invention means a saturated straight chain or branched chain alkyl group, preferably a chain alkyl group having 1 to 8, more preferably having-3 carbon atoms. Such as "C_1-3Typical examples of the straight or branched alkyl group "include methyl, ethyl, n-propyl, isopropyl.

The halogen in the invention comprises fluorine, chlorine, bromine and iodine. Preferred halogen groups are fluorine, chlorine or bromine.

The cycloalkyl groups described herein are saturated or unsaturated, substituted or unsubstituted cyclic groups, which rings may be monocyclic or fused, bridged or spiro rings or ring systems containing heteroatoms. Monocyclic ring is 3 to 12 atoms, preferably 3 to 9 atoms, more preferably 6 atoms, polycyclic ring is preferably 7 to 13 atoms, hetero atom means nitrogen, sulfur, phosphorus, oxygen, etc.

Alkoxy in the sense of the present invention means a radical-OR ', wherein R' is an alkyl radical as defined above in the sense of the present invention. Typical examples of "alkoxy" groups of the present invention include, but are not limited to, methoxy, ethoxy, n-propoxy, and the like.

Preferably, the compound may be further represented by formula (I), formula (II), formula (III), which is represented by the following structure:

wherein m is a positive integer from 1 to 6, for example m can be 1, 2,3, 4, 5 or 6;

y is O or S;

R¹、R²each independently selected from C_1-8Straight or branched chainChain alkyl radical, C_2-8Straight-chain or branched alkenyl, C_2-8Straight-chain or branched alkynyl, heteroalkyl, C_3-12Cycloalkyl, aryl, heteroaryl; said cycloalkyl, aryl, heteroaryl being unsubstituted or selected from halogen, nitro, amino, C_1-8Straight or branched alkyl, C_2-8Straight-chain or branched alkenyl, C_2-8One or more of straight chain or branched chain alkynyl, heteroalkyl and alkoxy.

Preferably, R¹Is selected from C_3-6Cycloalkyl, aryl, heteroaryl; said cycloalkyl, aryl, heteroaryl being unsubstituted or selected from halogen, nitro, amino, C_1-8Straight or branched alkyl, C_2-8Straight-chain or branched alkenyl, C_2-8One or more of straight chain or branched chain alkynyl, heteroalkyl and alkoxy.

Preferably, R¹Selected from cyclohexyl and phenyl; the cyclohexane or phenyl group may be substituted or selected from halogen, nitro, amino, C_1-3One or more of linear or branched alkyl, alkoxy; the halogen is selected from fluorine, chlorine, bromine and iodine.

Preferably, in the formula (I), R¹Is phenyl; said phenyl being unsubstituted or selected from halogen, C_1-3Straight or branched alkyl, C_1-3One or more of linear or branched alkoxy; the halogen is selected from fluorine, chlorine, bromine and iodine.

Preferably, in the formula (I), R¹When the phenyl is substituted, the position of the substituent is selected from C-3 and C-4. Preferably the C-4 position.

Preferably, in formula (II), m is a positive integer from 1 to 4, preferably 1 or 2; y is O or S.

Preferably, in formula (II), R²Selected from cyclohexyl and phenyl; the cyclohexane or phenyl group may be substituted or substituted by one or more selected from fluorine, chlorine, bromine, iodine.

Preferably, in formula (II), R²When substituted phenyl, the substitution is mono-or di-substituted.

PreferablyIn the formula (II), R²When the phenyl is substituted, the position of the substituent is selected from one or more of C-2, C-3 and C-4; preferably selected from the C-3 and C-4 positions.

Preferably, in formula (II), R²When substituted phenyl, the substituents are selected from fluorine, chlorine, bromine; chlorine is preferred.

Preferably, in formula (III), m is a positive integer from 1 to 4, preferably 1 or 2.

Preferably, in the formula (III), R¹Is phenyl; said phenyl being unsubstituted or selected from halogen, nitro, C_1-3One or more of linear or branched alkyl; the halogen is selected from fluorine, chlorine, bromine and iodine; bromine is preferred.

Preferably, in the formula (III), R¹When the phenyl is substituted, the position of the substituent is selected from C-3 and C-4.

Preferably, the compounds of the present invention are the following compounds:

1- ((3-bromophenyl) sulfonyl) -N- (5-nitrothiazol-2-yl) piperidine-4-carboxamide (compound I-1);

1- ((4-methoxyphenyl) sulfonyl) -N- (5-nitrothiazol-2-yl) piperidine-4-carboxamide (compound I-2);

1- ((4-chlorophenyl) sulfonyl) -N- (5-nitrothiazol-2-yl) piperidine-4-carboxamide (compound I-3);

1- ((4-fluorophenyl) sulfonyl) -N- (5-nitrothiazol-2-yl) piperidine-4-carboxamide (compound I-4);

n- (5-Nitrothiazol-2-yl) -1-tosylpiperidine-4-carboxamide (Compound I-5);

1- ((4-bromophenyl) sulfonyl) -N- (5-nitrothiazol-2-yl) piperidine-4-carboxamide (compound I-6);

n- (5-Nitrothiazol-2-yl) -4- (3-phenylsulfamoyl) butanamide (Compound II-1);

4- (3- (3-chlorophenyl) ureido) -N- (5-nitrothiazol-2-yl) butanamide (compound II-2);

4- (3-cyclohexylthioureido) -N- (5-nitrothiazol-2-yl) butanamide (compound II-3);

4- (3- (3-chloro-4-fluorophenyl) ureido) -N- (5-nitrothiazol-2-yl) butanamide (compound II-4);

4- (3- (2-bromophenyl) ureido) -N- (5-nitrothiazol-2-yl) butanamide (compound II-5);

4- (3- (4-chlorophenyl) ureido) -N- (5-nitrothiazol-2-yl) butanamide (compound II-6);

5- (3- (3-chloro-4-fluorophenyl) ureido) -N- (5-nitrothiazol-2-yl) pentanamide (Compound II-7);

5- (3- (4-chlorophenyl) ureido) -N- (5-nitrothiazol-2-yl) pentanamide (Compound II-8);

4- ((4-nitrophenyl) sulfonamido) -N- (5-nitrothiazol-2-yl) butanamide (compound III-1);

4- ((4-methylphenyl) sulfonamido) -N- (5-nitrothiazol-2-yl) butanamide (compound III-2);

4- ((4-bromophenyl) sulfonamido) -N- (5-nitrothiazol-2-yl) butanamide (compound III-3);

4- ((4-chlorophenyl) sulfonylamino) -N- (5-nitrothiazol-2-yl) butanamide (compound III-4);

4- ((3-bromophenyl) sulfonamido) -N- (5-nitrothiazol-2-yl) butanamide (compound III-5);

n- (5-Nitrothiazol-2-yl) -4- (phenylsulfonylamino) butanamide (Compound III-6);

5- ((4-bromophenyl) sulfonamido) -N- (5-nitrothiazol-2-yl) pentanamide (Compound III-7);

5- ((4-chlorophenyl) sulfonylamino) -N- (5-nitrothiazol-2-yl) pentanamide (Compound III-8);

n- (5-Nitrothiazol-2-yl) -5- (phenylsulfonylamino) pentanamide (Compound III-9);

5- ((3-bromophenyl) sulfonylamino) -N- (5-nitrothiazol-2-yl) pentanamide (Compound III-10).

The invention also includes pharmaceutically acceptable salts or prodrugs of the above compounds, and the invention also includes solvates of the above compounds. In addition, the invention also comprises various crystal forms of the compound.

The present invention provides pharmaceutically acceptable salts of the above compounds. Examples of the "pharmaceutically acceptable salt" according to the present invention include inorganic acid salts such as hydrochloride, hydrobromide, sulfate, phosphate and nitrate; organic acid salts such as acetate, propionate, oxalate, succinate, lactate, malate, tartrate, citrate, maleate, fumarate, methanesulfonate, benzenesulfonate, p-toluenesulfonate and ascorbate; inorganic base salts such as sodium salt, potassium salt, calcium salt, zinc salt, magnesium salt and aluminum salt; and organic base salts such as arginine salt, benzathine salt, choline salt, diethylamine salt, dialcohol amine salt, glycinate salt, lysine salt, meglumine salt, ethanolamine salt, and tromethamine salt.

The prodrug of the invention can react with enzyme, gastric acid and the like under physiological conditions in vivo so as to be converted into the compound; for example, the above-mentioned compounds of the present invention can be converted by oxidation, reduction, hydrolysis, etc. of an enzyme; and the above compounds of the present invention can be converted by hydrolysis using gastric acid.

The compounds of the invention may exist in tautomeric forms. Thus, the present invention also includes tautomers of the above compounds.

In a second aspect of the invention, the invention provides a process for the preparation of a compound of formula (V) comprising condensation of 2-amino-5-nitrothiazole as starting material with HOOC-A-Boc followed by de-Boc reaction with R¹-SO₂Cl or R²-NCHY reaction to obtain; wherein, A, R¹、R²Y is as defined above.

Preferably, the process for preparing the compound of formula (V) comprises carrying out the following reaction:

preferably, the above method comprises the steps of:

(1) 2-amino-5-nitrothiazole and HOOC-A-Boc are subjected to condensation reaction in a solvent A in the presence of an organic base, a condensing agent and an activating agent to generate an intermediate

(2) Intermediates

In an acid solution, reacting at room temperature to remove Boc protecting group to generate an intermediate

(3) Under the condition of acid-binding agent, the intermediate

After dissolving with solvent B and then neutralizing with R¹-SO₂Cl or R²-reacting the NCHY at room temperature to obtain a compound of formula (V);

wherein, A, R¹、R²Y is as defined above.

Preferably, in step (1), the solvent a is selected from anhydrous N, N-Dimethylformamide (DMF), tetrahydrofuran, acetonitrile, chloroform, dichloromethane, 1, 4-dioxane, preferably anhydrous dichloromethane and/or DMF;

preferably, in the step (1), the amount of the solvent is 20-25 mL per millimole of the solvent for 2-amino-5-nitrothiazole;

preferably, in step (1), the organic base is selected from triethylamine, 1, 8-diazabicycloundecen-7-ene (DBU), N-methylmorpholine (NMM), N-Diisopropylethylamine (DIPEA);

preferably, in step (1), the condensing agent is selected from Dicyclohexylcarbodiimide (DCC), Diisopropylcarbodiimide (DIC), 1- (3-dimethylaminopropyl) -3-Ethylcarbodiimide (EDCI), O- (7-azabenzotriazol-1-yl) -bis (dimethylamino) carbonium Hexafluorophosphate (HATU), O- (benzotriazol-1-yl) -bis (dimethylamino) carbonium Hexafluorophosphate (HBTU), O- (5-chlorobenzotriazol-1-yl) -bis (dimethylamino) carbonium Hexafluorophosphate (HCTU), O- (benzotriazol-1-yl) -bis (dimethylamino) carbonium tetrafluoroborate (tu), O- (N-succinimidyl) -bis (dimethylamino) carbonium tetrafluoroborate (TSTU), O- (N-endo-5-norcamphene-2, 3-dicarboximide) -bis (dimethylamino) carbonium tetrafluoroborate (TNTU), benzotriazol-1-yloxy-tris (tetrahydropyrrolyl) phosphonium hexafluorophosphate (PyBOP), preferably EDCI and/or HATU;

preferably, in the step (1), the activator is selected from the group consisting of 4-Dimethylaminopyridine (DMAP), 1-hydroxybenzotriazole (HOBt), 4-pyrrolidinylpyridine (4-PPY), 1-hydroxy-7-azobenzotriazol (HOAT), N-hydroxysuccinimide (HOSu), N-hydroxyphthalimide (NHPI), N-hydroxyphthalimide (NHNI), pentafluorophenol (PFPOH);

preferably, in the step (1), the molar ratio of the 2-amino-5-nitrothiazole, the HOOC-A-Boc, the condensing agent, the activating agent and the organic base is 1: 1.2-1.7: 1.2-1.7: 1-2.5: 1 to 3.0, preferably 1: 1.2: 1.2: 1-2.2: 1 to 2.5;

preferably, the reaction temperature of the step (1) is 0-60 ℃, preferably 30 ℃;

preferably, in step (2), the acidic solution is selected from the group consisting of HCl/ethyl acetate solution, HCl/methanol solution, trifluoroacetic acid, preferably HCl/ethyl acetate solution;

preferably, in step (2), the acidic solution is used in an amount per millimole of intermediate

10-15 mL of an acid solution is used;

preferably, in step (3), the reactant is R¹-SO₂When Cl is contained, the acid-binding agent is inorganic base, the inorganic base is selected from potassium carbonate, sodium carbonate or sodium bicarbonate, and potassium carbonate is preferred;

preferably, in step (3), the reactant is R¹-SO₂When Cl, the solvent B is a solvent/water system selected from dioxane/water, tetrahydrofuran/water and acetone/water; preferably, in the solvent/water system, the volume ratio of the solvent to the water is 1: 1-3; preferably, the solvent B is used in an amount per millimole of intermediate

The amount of the solvent B is 8-12 mL;

preferably, in step (3), the

Acid-binding agent and R₁-SO₂The molar ratio of Cl is 1: 2-4: 1 to 1.2, preferably 1: 3: 1.2;

preferably, in step (3), the reactant is R²-when NCHY, the acid scavenger is an organic base selected from triethylamine, DBU, NMM, DIPE, preferably triethylamine;

preferably, in step (3), the reactant is R²-NCHY, intermediate

The mol ratio of the acid binding agent to the acid binding agent is 1: 1 to 1.5, preferably 1: 1.2;

preferably, in step (3), the reactant is R²-NCHY, said solvent B is selected from anhydrous DMF, tetrahydrofuran, acetonitrile, chloroform, dichloromethane, 1, 4-dioxane, preferably anhydrous tetrahydrofuran;

preferably, in step (3), the reactant is R²-NCHY, the solvent is used in the following amount: per millimole of intermediate

The amount of the solvent B is 8-12 mL;

preferably, the intermediate

Acid-binding agent and R²-molar ratio of NCHY 1: 1-1.5: 1 to 1.5, preferably 1: 1.2: 1.2.

in the preparation method of the present invention, when the compound of formula (V) has the structure represented by formula (I), it is prepared by the following reaction:

wherein R is¹As defined hereinbefore.

Preferably, the process for the preparation of the compound of formula (I) comprises the steps of:

(1) carrying out condensation reaction on 2-amino-5-nitrothiazole and 1-Boc-4-piperidinecarboxylic acid in a solvent A in the presence of organic alkali, a condensing agent and an activating agent at 0-60 ℃, preferably 30 ℃ to generate an intermediate 1-1;

(2) the intermediate 1-1 reacts in an acid solution at room temperature to remove the Boc protecting group to generate an intermediate 1-2;

(3) under the condition of using inorganic base as acid-binding agent, the intermediate 1-2 is dissolved by solvent B and then is reacted with R¹Reacting substituted sulfonyl chloride at room temperature to obtain a compound shown in a formula (I);

preferably, in step (1), the solvent A is selected from DMF, tetrahydrofuran, acetonitrile, chloroform, dichloromethane, 1, 4-dioxane, preferably anhydrous dichloromethane and/or DMF;

preferably, in the step (1), the amount of the solvent A is 20-25 mL per millimole of the 2-amino-5-nitrothiazole;

preferably, in the step (1), the organic base is triethylamine, DBU, NMM, DIPEA, preferably triethylamine and/or NMM;

preferably, in step (1), the condensing agent is selected from DCC, DIC, EDCI, HATU, HBTU, HCTU, TBTU, TSTU, TNTU, PyBOP, preferably EDCI and/or HATU;

preferably, in step (1), the activator is selected from DMAP, HOBt, 4-PPY, HOAT, HOSu, NHPI, NHNI, PFPOH, preferably HOBt;

preferably, in the step (1), the molar ratio of the 2-amino-5-nitrothiazole to the 1-Boc-4-piperidinecarboxylic acid, the condensing agent, the activating agent and the organic base is 1: 1.2-1.7: 1.2-1.7: 1.2-1.7: 1-3, preferably 1: 1.2: 1.2: 1.2: 1.2;

preferably, in step (2), the acidic solution is selected from the group consisting of HCl/ethyl acetate solution, HCl/methanol solution, trifluoroacetic acid, preferably HCl/ethyl acetate solution;

preferably, in the step (2), the amount of the acidic solution is 10-15 mL per millimole of the intermediate 1-1;

preferably, in step (3), the inorganic base is selected from potassium carbonate, sodium bicarbonate, preferably potassium carbonate;

preferably, in step (3), the solvent B is a solvent/water system selected from dioxane/water, tetrahydrofuran/water, acetone/water; preferably, in the solvent/water system, the volume ratio of the solvent to the water is 1: 1-3;

preferably, in the step (3), the amount of the solvent B is 8-12 mL per millimole of the intermediate 1-2;

preferably, in the step (3), the intermediate 1-2, the inorganic base and the R¹The molar ratio of substituted sulfonyl chlorides was 1: 2-4: 1 to 1.5, preferably 1: 3: 1.2.

in the preparation method of the present invention, when the compound of formula (V) has the structure represented by formula (II), it is prepared by the following reaction:

wherein R is²M, Y are as defined above.

Preferably, the process for the preparation of the compound of formula (II) comprises the steps of:

(1) carrying out condensation reaction on 2-amino-5-nitrothiazole and N-Boc-gamma-aminobutyric acid or N-Boc-5-aminopentanoic acid in a solvent A in the presence of organic base, a condensing agent and an activating agent at 0-60 ℃, preferably 30 ℃ to generate an intermediate 2-1;

(2) the intermediate 2-1 is reacted in an acid solution at room temperature to remove a Boc protecting group to generate an intermediate 2-2;

(3) under the condition of organic base, dissolving the intermediate 2-2 with a solvent B, and neutralizing with R₂Substituted isocyanates or R₂(iii) a substituted isothiocyanate, reacted at room temperature to provide a compound of formula (II);

preferably, in step (1), the solvent A is selected from anhydrous DMF, tetrahydrofuran, acetonitrile, chloroform, dichloromethane, 1, 4-dioxane, preferably anhydrous dichloromethane and/or DMF;

preferably, in the step (1), the amount of the solvent A is 20-25 mL per millimole of the 2-amino-5-nitrothiazole;

preferably, in step (1), the organic base is selected from triethylamine, DBU, NMM, DIPEA, preferably DIPEA and/or NMM;

preferably, in step (1), the condensing agent is selected from DCC, DIC, EDCI, HATU, HBTU, HCTU, TBTU, TSTU, TNTU, PyBOP, preferably EDCI and/or HATU;

preferably, in step (1), the activator is selected from DMAP, HOBt, 4-PPY, HOAT, HOSu, NHPI, NHNI, PFPOH, preferably DMAP and/or HOBt;

preferably, in the step (1), the molar ratio of the 2-amino-5-nitrothiazole to the N-Boc-gamma-aminobutyric acid or the N-Boc-5-aminovaleric acid, the condensing agent, the activating agent and the organic base is 1: 1.2-1.7: 1.2-1.7: 1-2.5: 1-3.0, preferably 1: 1.2: 1.2: 2.2: 1.2;

preferably, in step (2), the acidic solution is selected from the group consisting of HCl/ethyl acetate solution, HCl/methanol solution, trifluoroacetic acid, preferably HCl/ethyl acetate solution;

preferably, in the step (2), the amount of the acidic solution is 10-15 mL per millimole of the intermediate 2-1;

preferably, in step (3), the organic base is selected from triethylamine, DBU, NMM, DIPEA, preferably triethylamine;

preferably, in the step (3), the molar ratio of the intermediate 2-2 to the organic base is 1: 1 to 1.5, preferably 1: 1.2;

preferably, in step (3), the solvent B is selected from anhydrous DMF, tetrahydrofuran, acetonitrile, chloroform, dichloromethane, 1, 4-dioxane, and preferably anhydrous tetrahydrofuran;

preferably, in the step (3), the amount of the solvent B is 8-12 mL per millimole of the intermediate 2-2;

preferably, in step (3), the intermediate 2-2, the organic base, R²Substituted isocyanates or R²The molar ratio of substituted isothiocyanates is 1: 1E1.5: 1 to 1.5, preferably 1: 1.2: 1.2.

in the preparation method of the present invention, when the compound of formula (V) has the structure represented by formula (III), it is prepared by the following reaction:

wherein R is¹M is as defined above.

Preferably, the process for the preparation of the compound of formula (III) comprises the steps of:

(1) carrying out condensation reaction on 2-amino-5-nitrothiazole and N-Boc-gamma-aminobutyric acid or N-Boc-5-aminopentanoic acid in a solvent A in the presence of organic base, a condensing agent and an activating agent at 0-60 ℃, preferably 30 ℃ to generate an intermediate 3-1;

(2) the intermediate 3-1 is reacted in an acid solution at room temperature to remove a Boc protecting group to generate an intermediate 3-2;

(3) under the condition that organic base is used as an acid-binding agent, the intermediate 3-2 is dissolved by a solvent B and then is mixed with R¹Reacting the substituted sulfonyl chloride at room temperature to obtain a compound of a formula (III);

preferably, in step (1), the solvent A is selected from anhydrous DMF, tetrahydrofuran, acetonitrile, chloroform, dichloromethane, 1, 4-dioxane, preferably anhydrous dichloromethane and/or DMF;

preferably, in the step (1), the amount of the solvent A is 20-25 mL per millimole of the 2-amino-5-nitrothiazole;

preferably, in step (1), the organic base is selected from triethylamine, DBU, NMM, DIPEA, preferably DIPEA and/or NMM;

preferably, in step (1), the condensing agent is selected from DCC, DIC, EDCI, HATU, HBTU, HCTU, TBTU, TSTU, TNTU, PyBOP, preferably EDCI and/or HATU;

preferably, in step (1), the activator is selected from DMAP, HOBt, 4-PPY, HOAT, HOSu, NHPI, NHNI, PFPOH, preferably DMAP and/or HOBt;

preferably, in the step (1), the molar ratio of the 2-amino-5-nitrothiazole to the N-Boc-gamma-aminobutyric acid or the N-Boc-5-aminovaleric acid, the condensing agent, the activating agent and the organic base is 1: 1.2-1.7: 1.2-1.7: 1.0-2.5: 1.0 to 3.0, preferably 1: 1.2: 1.2: 2.2: 1.2;

preferably, in step (2), the acidic solution is selected from the group consisting of HCl/ethyl acetate solution, HCl/methanol solution, trifluoroacetic acid, preferably HCl/ethyl acetate solution;

preferably, in the step (2), the amount of the acidic solution is 10-15 mL per millimole of the intermediate 3-1;

preferably, in step (3), the inorganic base is selected from potassium carbonate, sodium carbonate or sodium bicarbonate, preferably potassium carbonate;

preferably, in step (3), the solvent B is a solvent/water system selected from dioxane/water, tetrahydrofuran/water, acetone/water; preferably, in the solvent/water system, the volume ratio of the solvent to water is 1: 1-3;

preferably, in the step (3), the amount of the solvent B is 8-12 mL per millimole of the intermediate 3-2;

preferably, in step (3), the intermediate 3-2, inorganic base, R¹The molar ratio of substituted sulfonyl chlorides was 1: 2-4: 1 to 1.5, preferably 1: 3: 1.2.

in a third aspect of the invention, the invention provides a composition comprising a compound as described above or an isomer, solvate, prodrug and a pharmaceutically acceptable salt thereof. The inclusion is understood to mean that the above-mentioned compounds or isomers, solvates, prodrugs and pharmaceutically acceptable salts thereof, function as an adjunct ingredient in the compositions or as a primary active ingredient.

Preferably, the composition may comprise a pharmaceutically acceptable carrier, adjuvant and/or excipient.

In a fourth aspect of the present invention, there is provided a pharmaceutical preparation, food, cosmetic or cleaning product comprising a compound as described above or an isomer, solvate, prodrug thereof, and a pharmaceutically acceptable salt thereof, and a pharmaceutically, comestibly or cosmetically acceptable carrier, adjuvant and/or vehicle;

preferably, the pharmaceutical preparation is a solid preparation, an external preparation, a spray or a liquid preparation;

preferably, the food, cosmetic or cleaning product is a chewing gum, a buccal tablet, a toothpaste, a mouthwash or a disinfectant.

The compounds of the present invention or pharmaceutical compositions containing them may be administered in unit dosage form. The administration form may be a liquid form, a solid form, a preparation for external use, a spray, or the like. The liquid dosage form can be true solution, colloid, microparticle, emulsion, or mixed suspension. Other dosage forms such as tablet, capsule, dripping pill, aerosol, pill, powder, solution, suspension, emulsion, granule, suppository, lyophilized powder for injection, clathrate, landfill, patch, liniment, etc.

The pharmaceutical combination or pharmaceutical preparation of the present invention may further comprise a conventional carrier, wherein the pharmaceutically acceptable carrier includes but is not limited to: ion exchangers, aluminum oxide, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffer substances such as phosphates, glycerol, sorbates, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulosic substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, beeswax, lanolin and the like. The carrier may be present in the pharmaceutical composition in an amount of 1% to 98% by weight, usually about 80% by weight. For convenience, the local anesthetic, preservative, buffer, etc. may be dissolved directly in the vehicle.

Oral tablets and capsules may contain excipients such as binding agents, for example syrup, acacia, sorbitol, tragacanth, or polyvinylpyrrolidone, fillers such as lactose, sucrose, corn starch, calcium phosphate, sorbitol, glycine, lubricants such as magnesium stearate, talc, polyethylene glycol, silica, disintegrants such as potato starch, or acceptable wetting agents such as sodium lauryl sulfate. The tablets may be coated by methods known in the art of pharmacy.

The oral liquid can be made into water and oil suspension, solution, emulsion, syrup, or dried product, and supplemented with water or other suitable medium before use. Such liquid preparations may contain conventional additives such as suspending agents, sorbitol, cellulose methyl ether, glucose syrup, gelatin, hydroxyethyl cellulose, carboxymethyl cellulose, aluminum stearate gelatin, hydrogenated edible fats and oils, emulsifying agents such as lecithin, sorbitan monooleate, gum arabic; or a non-aqueous carrier (which may comprise an edible oil), such as almond oil, an oil such as glycerol, ethylene glycol, or ethanol; preservatives, e.g. methyl or propyl p-hydroxybenzoates, sorbic acid. Flavoring or coloring agents may be added if desired.

In addition, the solid preparation can also be made into toothpaste, solid collutory, chewing gum, buccal tablet, oral patch, etc.; the spray or liquid preparation for external use can also be made into collutory, lotion, etc.

In a fifth aspect of the present invention, the present invention provides the use of the above compound or its isomer, solvate, prodrug, and pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing the same as an active ingredient for the preparation of a medicament, food, cosmetic, or cleaning product for inhibiting and/or killing oral cariogenic bacteria.

In a sixth aspect of the invention, the invention provides the application of the compound or the isomer, the solvate, the prodrug and the pharmaceutically acceptable salt thereof or the pharmaceutical composition taking the compound or the isomer, the solvate, the prodrug and the pharmaceutically acceptable salt thereof as active ingredients in the preparation of drugs, cosmetics or cleaning products for inhibiting the biofilm formation caused by the streptomyces mutans and/or the plaque organisms and/or in the preparation of drugs, cosmetics or cleaning products for clearing the biofilm caused by the streptomyces mutans.

In a seventh aspect of the invention, the invention provides the use of the above compound or its isomer, solvate, prodrug and pharmaceutically acceptable salt or a pharmaceutical composition containing the same as an active ingredient in the preparation of an oral mutans bacteria biofilm inhibitor.

In an eighth aspect of the present invention, the present invention provides the use of the above compound or its isomer, solvate, prodrug, and pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing the same as an active ingredient for preparing a pharmaceutical preparation for combating caries.

The results show that the compounds of the invention have a broad spectrum of bacteriostatic and bactericidal activity against oral bacteria. Especially the minimum inhibitory concentration to the viscous actinomycetes is as low as 0.5 mu g/mL; the minimum inhibitory concentration to the streptococcus mutans and the streptococcus salivarius is as low as 2 mu g/mL; the minimum bactericidal concentration to the streptomyces variabilis is as low as 4 mu g/mL; can effectively inhibit the formation of the variable chain bacteria biofilm, and the minimum MBIC value can reach 2 mug/mL; can obviously reduce the growth activity of mature biofilm of streptomyces variabilis, and the MBEC of the mature biofilm₅₀The value was 16. mu.g/mL. And the compound of the invention has obvious inhibition effect on the acid production of the streptomyces variabilis under MIC (4 mug/mL) and 1/2MIC (2 mug/mL). In addition, the compound has no influence on normal oral bacteria, and can selectively act on cariogenic bacteria in a biofilm.

Drawings

Embodiments of the invention are described in detail below with reference to the attached drawing figures, wherein:

FIG. 1 shows the effect of II-2 on the acid-producing ability of a mutan.

Detailed Description

The invention will be further illustrated with reference to the following specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. The experimental procedures, in which specific conditions are not noted in the following examples, are generally carried out according to conventional conditions or according to conditions recommended by the manufacturers.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. In addition, any methods and materials similar or equivalent to those described herein can be used in the methods of the present invention. The preferred embodiments and materials described herein are intended to be exemplary only.

The preparation method of the compound comprises the steps of taking 2-amino-5-nitrothiazole as a starting material, condensing with HOOC-A-Boc, removing Boc reaction, and reacting with R¹-SO₂Cl or R²-NCHY reaction to obtain; wherein, A, R¹、R²And Y is as defined herein. The reaction route is as follows:

in a more specific embodiment, the method includes the following steps:

(1) 2-amino-5-nitrothiazole and HOOC-A-Boc are subjected to condensation reaction in a solvent A in the presence of an organic base, a condensing agent and an activating agent to generate an intermediate

(2) Intermediates

In an acid solution, reacting at room temperature to remove Boc protecting group to generate an intermediate

(3) Under the condition of acid-binding agent, the intermediate

After dissolving with solvent B and then neutralizing with R¹-SO₂Cl or R²-reacting the NCHY at room temperature to obtain a compound of formula (V);

wherein, A, R¹、R²Y is as defined above.

In a more preferred embodiment, in step (1), the solvent a is selected from anhydrous N, N-Dimethylformamide (DMF), tetrahydrofuran, acetonitrile, chloroform, dichloromethane, 1, 4-dioxane;

in a more preferred embodiment, in the step (1), the amount of the solvent is 20 to 25mL per millimole of the solvent for 2-amino-5-nitrothiazole;

in a more preferred embodiment, in step (1), the organic base is selected from triethylamine, 1, 8-diazabicycloundecen-7-ene (DBU), N-methylmorpholine (NMM), N-Diisopropylethylamine (DIPEA);

in a more preferred embodiment, in step (1), the condensing agent is selected from Dicyclohexylcarbodiimide (DCC), Diisopropylcarbodiimide (DIC), 1- (3-dimethylaminopropyl) -3-Ethylcarbodiimide (EDCI), O- (7-azabenzotriazol-1-yl) -bis (dimethylamino) carbonium Hexafluorophosphate (HATU), O- (benzotriazol-1-yl) -bis (dimethylamino) carbonium Hexafluorophosphate (HBTU), O- (5-chlorobenzotriazol-1-yl) -bis (dimethylamino) carbonium Hexafluorophosphate (HCTU), O- (benzotriazol-1-yl) -bis (dimethylamino) carbonium tetrafluoroborate (TBTU), O- (N-succinimidyl) -bis (dimethylamino) carbonium tetrafluoroborate (TSTU) ) O- (N-endo-5-norbornene-2, 3-dicarboximide) -bis (dimethylamino) carbonium tetrafluoroborate (TNTU), benzotriazol-1-yloxy-tris (tetrahydropyrrolyl) phosphonium hexafluorophosphate (PyBOP);

in a more preferred embodiment, in step (1), the activator is selected from the group consisting of 4-Dimethylaminopyridine (DMAP), 1-hydroxybenzotriazole (HOBt), 4-pyrrolidinylpyridine (4-PPY), 1-hydroxy-7-azobenzotriazol (HOAT), N-hydroxysuccinimide (HOSu), N-hydroxyphthalimide (NHPI), N-hydroxyphthalimide (NHNI), pentafluorophenol (PFPOH);

in a more preferred embodiment, in step (1), the 2-amino-5-nitrothiazole, the HOOC-a-Boc, the condensing agent, the activating agent, and the organic base are used in a molar ratio of 1: 1.2-1.7: 1.2-1.7: 1-2.5: 1 to 3.0, preferably 1: 1.2: 1.2: 1-2.2: 1 to 2.5;

in a more preferred embodiment, the reaction temperature in step (1) is 0-60 ℃, preferably 30 ℃;

in a more preferred embodiment, in step (2), the acidic solution is selected from the group consisting of HCl/ethyl acetate solution, HCl/methanol solution, trifluoroacetic acid;

in a more preferred embodiment, in step (2), the acidic solution is used in an amount per millimole of intermediate

10-15 mL of an acid solution is used;

in a more preferred embodiment of the present invention,in the step (3), the reactant is R¹-SO₂When Cl is contained, the acid-binding agent is inorganic base, and the inorganic base is selected from potassium carbonate, sodium carbonate or sodium bicarbonate;

in a more preferred embodiment, in step (3), the reactant is R¹-SO₂When Cl, the solvent B is a solvent/water system selected from dioxane/water, tetrahydrofuran/water and acetone/water; preferably, in the solvent/water system, the volume ratio of the solvent to the water is 1: 1-3; preferably, the solvent B is used in an amount per millimole of intermediate

The amount of the solvent B is 8-12 mL;

in a more preferred embodiment, in the step (3), the step (c) is performed by

Acid-binding agent and R₁-SO₂The molar ratio of Cl is 1: 2-4: 1 to 1.2, preferably 1: 3: 1.2;

in a more preferred embodiment, in step (3), the reactant is R²-when NCHY, the acid scavenger is an organic base selected from triethylamine, DBU, NMM, DIPE;

in a more preferred embodiment, in step (3), the reactant is R²-NCHY, intermediate

The mol ratio of the acid binding agent to the acid binding agent is 1: 1 to 1.5, preferably 1: 1.2;

in a more preferred embodiment, in step (3), the reactant is R²-NCHY, said solvent B is selected from anhydrous DMF, tetrahydrofuran, acetonitrile, chloroform, dichloromethane, 1, 4-dioxane;

in a more preferred embodiment, in step (3), the reactant is R²-NCHY, the solvent is used in the following amount: per millimole of intermediate

The amount of the solvent B is 8-12 mL;

in a more preferred embodiment, the intermediate

Acid-binding agent and R²-molar ratio of NCHY 1: 1-1.5: 1 to 1.5, preferably 1: 1.2: 1.2.

synthesis of exemplary Compounds of formula (I)

Example 1: preparation method of 1- ((3-bromophenyl) sulfonyl) -N- (5-nitrothiazol-2-yl) piperidine-4-formamide (compound I-1)

Preparation of 4- ((5-nitrothiazol-2-yl) carbamoyl) piperidine-1-carboxylic acid tert-butyl ester (1-1) intermediate

1-Boc-4-piperidinecarboxylic acid (1.2mmol) was dissolved in 25mL of anhydrous N, N-dimethylformamide, HOBt (1.2mmol) and EDCI (1.2mmol) were added, respectively, and triethylamine (1.2mmol) was added dropwise. After the completion of the dropwise addition, 2-amino-5-nitrothiazole (1.0mmol) was added thereto, and reacted at 30 ℃ overnight, 200mL of distilled water was added to the reaction solution, the aqueous phase was extracted 3 times (3X 200mL) with ethyl acetate, the combined organic phases were washed 1 time (1X 100mL) with a saturated sodium chloride solution, dried over anhydrous magnesium sulfate, filtered under reduced pressure, and then evaporated and concentrated with a rotary evaporator to give a crude product. The crude product was isolated by column chromatography on silica gel (dichloromethane: methanol ═ 200:1) to give a yellow solid in 75% yield.

Preparation of intermediate N- (5-nitrothiazol-2-yl) piperidine-4-carboxamide hydrochloride (1-2)

Slowly dropping acetyl chloride into absolute ethyl alcohol (V: V ═ 4:5), introducing generated HCl gas into ethyl acetate to prepare HCl ethyl acetate saturated solution, dissolving the intermediate in the previous step into the HCl ethyl acetate saturated solution, stirring the solution at normal temperature for 15 minutes, detecting the reaction by TLC completely, evaporating the solvent to dryness to obtain yellow solid, wherein the yield of the crude product is 100%, and the crude product is directly used in the next step without treatment.

Preparation of (3) 1- ((3-bromophenyl) sulfonyl) -N- (5-nitrothiazol-2-yl) piperidine-4-carboxamide (I-1)

Intermediate 1-2(1mmol) and potassium carbonate (1: (1mmol) at room temperature3mmol) was dissolved in 8mL dioxane aqueous solution, m-bromobenzenesulfonyl chloride (1.2mmol) was added, stirred for 30min, and the reaction was monitored by TLC until the starting material was completely reacted. Dioxane was distilled off, 25mL of distilled water was added to the reaction solution, the aqueous phase was extracted 3 times with ethyl acetate (3X 25mL), the organic phases were combined, washed 1 time with saturated sodium chloride solution (1X 25mL), dried over anhydrous magnesium sulfate, filtered under reduced pressure, and then concentrated by vacuum evaporation to give the crude product. The crude product was purified and separated by silica gel chromatography (200-300 mesh) using 80:1 dichloromethane to give the desired product as a yellow solid in 65% yield.¹H NMR(400MHz,DMSO-d₆)：δ13.11(s,1H),8.63(s,1H),7.97(d,J＝8.0Hz,1H),7.90(s,1H),7.79(d,J＝8.0Hz,1H),7.64(t,J＝7.9Hz,1H),3.69(d,J＝12.0Hz,1H),2.58(m,1H),2.41(t,J＝11.9Hz,2H),1.96(d,J＝13.3Hz,2H),1.64(m,2H)ppm；ESI-MS:473.8[M-H]^-。

Example 2: preparation method of 1- ((4-methoxyphenyl) sulfonyl) -N- (5-nitrothiazol-2-yl) piperidine-4-formamide (I-2)

The preparation method is the same as I-1, and the yield is 61%.¹H NMR(400MHz,DMSO-d₆)：δ13.10(s,1H),8.63(s,1H),7.70(d,J＝8.5Hz,2H),7.18(d,J＝8.5Hz,2H),3.87(s,3H),3.63(d,J＝12.0Hz,1H),2.29(m,2H),1.95(d,J＝13.1Hz,2H),1.65(m,2H)ppm；ESI-MS:427.0[M-H]^-.

Example 3: preparation method of 1- ((4-chlorphenyl) sulfonyl) -N- (5-nitrothiazole-2-yl) piperidine-4-formamide (I-3)

The preparation method is the same as I-1, and the yield is 63%.¹H NMR(400MHz,DMSO-d₆)：δ13.12(s,1H),8.63(s,1H),7.79(d,J＝8.6Hz,2H),7.74(d,J＝8.6Hz,2H),3.66(d,J＝12.1Hz,1H),2.57(m,1H),2.38(t,J＝11.9Hz,2H),1.93(d,J＝3.7Hz,1H),1.66(m,2H)ppm；ESI-MS:430.1[M-H]^-.

Example 4: preparation method of 1- ((4-fluorophenyl) sulfonyl) -N- (5-nitrothiazol-2-yl) piperidine-4-formamide (I-4)

The preparation method is the same as I-1, and the yield is 59%.¹H NMR(400MHz,DMSO-d₆)：δ13.11(s,1H),8.63(s,1H),7.85(m,2H),7.51(t,J＝8.7Hz,2H),3.66(d,J＝12.1Hz,1H),2.56(m,1H),2.36(m,2H),1.95(d,J＝13.8Hz,2H),1.65(m,2H)ppm；ESI-MS:414.0[M-H]^-.

Example 5: preparation method of N- (5-nitrothiazol-2-yl) -1-tosylpiperidine-4-formamide (I-5)

The preparation method is the same as I-1, and the yield is 69%.¹H NMR(400MHz,DMSO-d₆)：δ13.09(s,1H),8.63(s,1H),7.65(d,J＝8.0Hz,2H),7.47(d,J＝8.0Hz,2H),3.63(d,J＝11.6Hz,2H),2.55(m,1H),2.42(s,3H),2.30(t,J＝11.9Hz,2H),1.94(d,J＝13.7Hz,2H),1.65(m,2H)ppm；ESI-MS:411.1[M+H]⁺.

Example 6: preparation method of 1- ((4-bromophenyl) sulfonyl) -N- (5-nitrothiazol-2-yl) piperidine-4-formamide (I-6)

The preparation method is the same as I-1, and the yield is 65%.¹H NMR(400MHz,DMSO-d₆)：δ13.12(s,1H),8.63(s,1H),7.88(d,J＝8.5Hz,2H),7.70(d,J＝8.6Hz,2H),3.66(d,J＝12.1Hz,1H),2.58(m,1H),2.38(t,J＝11.6Hz,2H),1.95(d,J＝13.5Hz,2H),1.65(m,2H)ppm；ESI-MS:474.0[M-H]^-.

Example 7: preparation method of 1- ((4-nitrophenyl) sulfonyl) -N- (5-nitrothiazol-2-yl) piperidine-4-formamide (I-7)

The preparation method is the same as I-1, and the yield is 70%.¹H NMR(400MHz,DMSO-d₆)：δ13.03(s,1H),8.61(s,1H),8.42(d,J＝8.9Hz,2H),8.06(t,J＝5.8Hz,1H),8.03(d,J＝8.7Hz,2H),3.66(d,J＝12.1Hz,1H),2.57(m,1H),2.38(t,J＝11.9Hz,2H),1.93(d,J＝13.5Hz,2H),1.66(m,2H)ppm；ESI-MS:440.04[M-H]^-。

Synthesis of exemplary Compounds of formula (II)

Example 8: preparation method of N- (5-nitrothiazol-2-yl) -4- (3-phenylthioureido) butanamide (compound II-1)

Preparation of (4- ((5-nitrothiazol-2-yl) amino) -4-oxobutyl) carbamic acid tert-butyl ester (2-1) as intermediate (1).

Dissolving N-Boc-gamma-aminobutyric acid in 25mL of anhydrous dichloromethane, sequentially adding EDCI (1.2mmol), HOBT (1.2mmol), DMAP (1mmol) and N, N-diisopropylethylamine (1.2mmol) into the reaction solution at room temperature, stirring for reacting for 30min, adding 5-nitro-2-aminothiazole (1mmol) for continuing for 11h, detecting the reaction by TLC, and stopping the reaction if no raw material is left. The reaction mixture was washed 2 times (2X 100mL) with 100mL of citric acid solution, then 1 time (1X 100mL) with saturated sodium bicarbonate solution, then 1 time (1X 100mL) with saturated sodium chloride solution, dried over anhydrous magnesium sulfate, filtered under reduced pressure, and then concentrated by evaporation to give the crude product. The crude product was isolated by silica gel column purification (dichloromethane: methanol ═ 200:1) to give a yellow solid in 80% yield.

Preparation of 4-amino-N- (5-nitrothiazol-2-yl) butanamide hydrochloride (2-2) as intermediate in < 2 >

Slowly dropping acetyl chloride into absolute ethyl alcohol (V: V ═ 4:5), introducing generated HCl gas into ethyl acetate to prepare HCl-ethyl acetate saturated solution, dissolving the intermediate in the previous step into the solution, stirring the solution at normal temperature for 15 minutes, detecting the reaction by TLC completely, evaporating the solvent to dryness to obtain yellow solid, wherein the yield of the crude product is 100%, and the crude product is directly used for the next reaction without further purification treatment.

Preparation of < 3 > N- (5-nitrothiazol-2-yl) -4- (3-phenylthioureido) butanamide (II-1)

Dissolving the intermediate 2-2(1eq) and triethylamine (1.2eq) in 8mL of anhydrous tetrahydrofuran at 0 ℃, dropwise adding phenyl isothiocyanate (1.2eq) again, stirring for 30min, detecting the reaction by TLC, distilling off tetrahydrofuran, adding 50mL of distilled water into the reaction solution, extracting the aqueous phase with ethyl acetate for 3 times (3X 50mL), combining the organic phases, washing with saturated sodium chloride solution for 1 time (1X 50mL), drying with anhydrous magnesium sulfate, filtering under reduced pressure, and then concentrating by vacuum evaporation to obtain a crude product. The crude product was purified by silica gel chromatography (200-300 mesh) using dichloromethane: methanol 50:1 as eluent to afford the desired product as a yellow solid in 77% yield.¹H NMR(400MHz,DMSO-d₆)：δ13.09(s,1H),9.50(s,1H),8.63(s,1H),7.79(s,1H),7.38(d,J＝7.5Hz,2H),7.32(t,J＝7.7Hz,2H),7.11(t,J＝7.2Hz,1H),3.53(d,J＝7.2Hz,2H),2.59(t,J＝7.3Hz,2H),1.91(m,2H)ppm；ESI-MS:364.0[M-H]^-.

Example 9: preparation method of 4- (3- (3-chlorphenyl) ureido) -N- (5-nitrothiazole-2-yl) butanamide (II-2)

The preparation method is the same as II-1, and the yield is 84%.¹H NMR(400MHz,DMSO-d₆)：δ13.06(s,1H),8.66(s,1H),8.58(s,1H),7.64(s,1H),7.21(t,J＝8.0Hz,1H),7.14(d,J＝8.4Hz,1H),6.91(d,J＝7.2Hz,1H),6.25(t,J＝5.9Hz,1H),3.14(q,J＝6.5Hz,2H),2.57(t,J＝7.2Hz,2H),1.80(m,2H)ppm；ESI-MS:382.0[M-H]^-.

Example 10: preparation method of 4- (3-cyclohexyl thiourea) -N- (5-nitrothiazole-2-yl) butanamide (II-3)

The preparation method is the same as II-1, and the yield is 37.6%.¹H NMR(400MHz,DMSO-d₆)：δ13.08(s,1H),8.63(s,1H),7.27(d,J＝32.6Hz,2H),3.91(s,1H),2.56(t,J＝7.3Hz,2H),1.85(m,4H),1.65(d,J＝12.2Hz,2H),1.55(d,J＝12.1Hz,1H),1.19(m,6H)ppm；ESI-MS:369.7[M-H]^-.

Example 11: preparation method of 4- (3- (3-chloro-4-fluorophenyl) ureido) -N- (5-nitrothiazol-2-yl) butanamide (II-4)

The preparation method is the same as II-1, and the yield is 83 percent.¹H NMR(400MHz,DMSO-d₆)：δ13.06(s,1H),8.66(s,1H),8.58(s,1H),7.73(dd,J＝6.8,2.5Hz,1H),7.24(t,J＝9.0Hz,1H),7.18(m,1H),6.25(t,J＝5.8Hz,1H),3.14(q,J＝6.5Hz,2H),2.57(t,J＝7.2Hz,2H),1.81(m,2H)ppm；ESI-MS:400.0[M-H]^-.

Example 12: preparation method of 4- (3- (2-bromophenyl) ureido) -N- (5-nitrothiazole-2-yl) butanamide (II-5)

The preparation method is the same as II-1, and the yield is 79%.¹H NMR(400MHz,DMSO-d₆)：δ13.08(s,1H),8.60(s,1H),8.05(d,J＝8.3Hz,1H),7.78(s,1H),7.53(d,J＝8.0Hz,1H),7.26(t,J＝7.2Hz,1H),7.14(t,J＝5.6Hz,1H),6.88(td,J＝7.7,1.6Hz,1H),3.16(q,J＝6.4Hz,2H),2.59(t,J＝7.3Hz,2H),1.81(m,2H)ppm；ESI-MS:425.8[M-H]^-.

Example 13: preparation method of 4- (3- (4-chlorphenyl) ureido) -N- (5-nitrothiazole-2-yl) butanamide (II-6)

The preparation method is the same as II-1, and the yield is 61.5%.¹H NMR(400MHz,DMSO-d₆)：δ13.06(s,1H),8.62(s,1H),8.57(s,1H),7.41(d,J＝8.9Hz,2H),7.25(d,J＝8.9Hz,2H),6.20(t,J＝5.7Hz,1H),3.14(q,J＝6.5Hz,2H),2.57(t,J＝7.2Hz,2H),1.80(m,2H)ppm；ESI-MS:381.9[M-H]^-.

Example 14: preparation method of 5- (3- (3-chloro-4-fluorophenyl) ureido) -N- (5-nitrothiazol-2-yl) pentanamide (II-7)

The preparation method is the same as II-1, and the yield is 61.8%.¹H NMR(400MHz,DMSO-d₆)：δ13.07(s,1H),8.63(d,J＝6.2Hz,2H),7.75(dd,J＝6.9,2.6Hz,1H),7.26(t,J＝9.0Hz,1H),7.20(m,1H),6.26(t,J＝5.7Hz,1H),3.09(q,J＝6.5Hz,2H),2.56(t,J＝7.4Hz,2H),1.64(m,2H),1.46(m,2H)ppm；ESI-MS:416.1[M+H]⁺.

Example 15: preparation method of 5- (3- (4-chlorphenyl) ureido) -N- (5-nitrothiazole-2-yl) pentanamide (II-8)

The preparation method is the same as II-1, and the yield is 49.3%.¹H NMR(400MHz,DMSO-d₆)：δ13.07(s,1H),8.63(s,1H),8.57(s,1H),7.41(d,J＝8.9Hz,2H),7.25(d,J＝8.9Hz,2H),6.20(t,J＝5.7Hz,1H),3.09(q,J＝6.5Hz,2H),2.56(t,J＝7.4Hz,2H),1.64(m,2H),1.46(m,2H)ppm；ESI-MS:396.0[M-H]^-.

Synthesis of exemplary Compounds of formula (III)

Example 16: preparation method of 4- ((4-nitrophenyl) sulfonamido) -N- (5-nitrothiazol-2-yl) butanamide (III-1)

Preparation of (4- ((5-nitrothiazol-2-yl) amino) -4-oxobutyl) carbamic acid tert-butyl ester (3-1) as intermediate

Dissolving N-Boc-gamma-aminobutyric acid in 25mL of anhydrous dichloromethane, sequentially adding EDCI (1.2eq), HOBT (1.2eq), DMAP (1eq) and N, N-diisopropylethylamine (1.2eq) into a reaction solution at room temperature, stirring for reacting for 30min, adding 5-nitro-2-aminothiazole (1eq) for continuously reacting for 11h, detecting by TLC, and stopping the reaction if no raw materials remain. The reaction mixture was washed 2 times (2X 100mL) with 100mL of citric acid solution, then 1 time (1X 100mL) with saturated sodium bicarbonate solution, then 1 time (1X 100mL) with saturated sodium chloride solution, dried over anhydrous magnesium sulfate, filtered under reduced pressure, and then concentrated by evaporation to give the crude product. The crude product was isolated by silica gel column purification (dichloromethane: methanol ═ 100:1) to give a yellow solid in 80% yield.

Preparation of 4-amino-N- (5-nitrothiazol-2-yl) butanamide hydrochloride (3-2) as intermediate in < 2 >

Slowly dropping acetyl chloride into absolute ethyl alcohol (V: V ═ 4:5), directly introducing generated HCl gas into ethyl acetate to prepare HCl/ethyl acetate saturated solution, dissolving the intermediate in the previous step into the saturated solution, stirring the saturated solution at normal temperature for 15 minutes, detecting the reaction by TLC (thin layer chromatography), evaporating the solvent to dryness to obtain yellow solid, wherein the yield of the crude product is 100%, and directly carrying out the next step without treatment.

Preparation of (3) 4- ((4-nitrophenyl) sulfonylamino) -N- (5-nitrothiazol-2-yl) butanamide (III-1)

At room temperature, the intermediate 3-2(1eq) and potassium carbonate (3eq) were dissolved in 8mL of dioxane aqueous solution, p-nitrobenzenesulfonyl chloride (1.2eq) was added, stirred for 30min, and the reaction was monitored by TLC until the starting material was completely reacted. Dioxane was distilled off, 25mL of distilled water was added to the reaction solution, the aqueous phase was extracted 3 times (3X 25mL) with dichloromethane, the organic phases were combined, washed 1 time (1X 25mL) with saturated sodium chloride solution, dried over anhydrous magnesium sulfate, filtered under reduced pressure, and then concentrated by vacuum evaporation to give the crude product. The crude product was purified by silica gel chromatography (200-300 mesh) using dichloromethane-methanol 80:1 as eluent to afford the desired product as a yellow solid in 63.7% yield.¹H NMR(400MHz,DMSO-d₆)：δ13.03(s,1H),8.61(s,1H),8.41(d,J＝8.9Hz,2H),8.07(t,J＝5.8Hz,1H),8.03(d,J＝8.7Hz,2H),2.87(q,J＝6.6Hz,2H),2.55(t,J＝7.2Hz,2H),1.74(m,2H)ppm；ESI-MS:434.0[M-H]^-.

Example 17: preparation method of 4- ((4-methylphenyl) sulfonylamino) -N- (5-nitrothiazol-2-yl) butanamide (III-2)

The preparation method is the same as III-1, and the yield is 70%.¹H NMR(400MHz,DMSO-d₆)：δ13.04(s,1H),8.62(s,1H),7.66(d,J＝8.0Hz,2H),7.58(t,J＝5.9Hz,1H),7.38(d,J＝8.0Hz,2H),2.76(q,J＝6.7Hz,2H),2.55(t,J＝7.2Hz,2H),2.37(s,3H),1.72(m,2H)ppm；ESI-MS:382.6[M-H]^-.

Example 18: preparation method of 4- ((4-bromophenyl) sulfonylamino) -N- (5-nitrothiazol-2-yl) butanamide (III-3)

The preparation method is the same as III-1, and the yield is 81 percent.¹H NMR(400MHz,DMSO-d₆)：δ13.04(s,1H),8.62(s,1H),7.80(m,3H),7.71(d,J＝8.5Hz,2H),2.80(q,J＝6.6Hz,2H),2.56(t,J＝7.2Hz,2H),1.72(m,2H)ppm；ESI-MS:447.3[M-H]^-.

Example 19: preparation method of 4- ((4-chlorphenyl) sulfonylamino) -N- (5-nitrothiazole-2-yl) butanamide (III-4)

The preparation method is the same as III-1, and the yield is 70%.¹H NMR(400MHz,DMSO-d₆)：δ13.05(s,1H),8.63(s,1H),7.79(m,3H),7.67(d,J＝8.5Hz,2H),2.80(q,J＝6.6Hz,2H),2.56(t,J＝7.2Hz,2H),1.73(m,2H)ppm；ESI-MS:404.7[M-H]^-.

Example 20: preparation method of 4- ((3-bromophenyl) sulfonylamino) -N- (5-nitrothiazol-2-yl) butanamide (III-5)

The preparation method is the same as III-1, and the yield is 70%.¹H NMR(400MHz,DMSO-d₆)：δ13.06(s,1H),8.63(s,1H),7.92(s,1H),7.86(m,2H),7.79(d,J＝8.0,1.2Hz,1H),7.57(t,J＝7.9Hz,1H),2.82(q,J＝6.7Hz,2H),2.57(t,J＝7.2Hz,2H),1.74(m,2H)ppm；ESI-MS:448.0[M-H]^-.

Example 21: preparation method of N- (5-nitrothiazole-2-yl) -4- (phenylsulfonylamino) butanamide (III-6)

The preparation method is the same as III-1, and the yield is 67.5%.¹H NMR(400MHz,DMSO-d₆)：δ13.06(s,1H),8.63(s,1H),7.79(d,J＝6.8Hz,1H),7.69(t,J＝5.9Hz,1H),7.61(m,3H),2.79(q,J＝6.7Hz,2H),2.56(t,J＝7.2Hz,2H),1.72(m,2H)ppm；ESI-MS:369.6[M-H]^-.

Example 22: preparation method of 5- ((4-bromophenyl) sulfonylamino) -N- (5-nitrothiazol-2-yl) pentanamide (III-7)

The preparation method is the same as III-1, and the yield is 58%.¹H NMR(400MHz,DMSO-d₆)：¹H NMR(400MHz,DMSO-d₆)：δ13.03(s,1H),8.63(s,1H),7.79(d,J＝8.6Hz,2H),7.73(t,J＝5.9Hz,1H),7.66(d,J＝8.6Hz,2H),2.76(q,J＝6.7Hz,2H),2.47(m,2H),1.58(m,2H),1.40(m,2H)ppm；ESI-MS:416.9[M-H]^-.

Example 23: preparation method of 5- ((4-chlorphenyl) sulfonylamino) -N- (5-nitrothiazole-2-yl) pentanamide (III-8)

Preparation methodThe method is the same as III-1, and the yield is 66.7%.¹H NMR(400MHz,DMSO-d₆)：δ13.03(s,1H),8.63(s,1H),7.79(d,J＝8.6Hz,2H),7.73(t,J＝5.9Hz,1H),7.66(d,J＝8.6Hz,2H),2.76(q,J＝6.7Hz,2H),2.47(m,2H),1.58(m,2H),1.40(m,2H)ppm；ESI-MS:419.3[M+H]⁺.

Example 24: preparation method of N- (5-nitrothiazole-2-yl) -5- (phenylsulfonylamino) pentanamide (III-9)

The preparation method is the same as III-1, and the yield is 72.7%.¹H NMR(400MHz,DMSO-d₆)：δ13.03(s,1H),8.63(s,1H),7.79(d,J＝7.2Hz,2H),7.61(m,4H),2.75(q,J＝6.7Hz,2H),2.47(m,2H),1.58(m,2H),1.40(m,2H)ppm；ESI-MS:382.8[M-H]^-.

Example 25: preparation method of 5- ((3-bromophenyl) sulfonylamino) -N- (5-nitrothiazol-2-yl) pentanamide (III-10)

The preparation method is the same as III-1, and the yield is 65.4%.¹H NMR(400MHz,DMSO-d₆)：δ13.04(s,1H),8.63(s,1H),7.92(s,1H),7.85(d,J＝8.0Hz,1H),7.79(m,2H),7.56(t,J＝7.9Hz,1H),2.78(q,J＝6.7Hz,2H),2.48(m,2H),1.59(m,2H),1.41(m,2H)ppm；ESI-MS:460.8[M-H]^-.

Example 26: antibacterial activity test experiment and test result of compound related to the invention

The activity test method is exemplified by using the variant streptococci UA159 strain as an example.

1. Preparation of Strain of mutans bacterium UA159 and Compounds

The mutant strain UA159 used in the present invention is a model strain, and the reference genome number in NCBI database (http:// www.ncbi.nlm.nih.gov /) is NC-004350. The mutans UA246 strain used in the present invention is a clinical strain isolated from the oral cavity of a patient suffering from dental caries. The most suitable medium is preferably Brain Heart Infusion (Brain Heart Infusion) medium (Brain Infusion solids 12.5g/L, Beef Heart Infusion solids5.0g/L, protein peptone 10.0g/L, Glucose 2.0g/L, Sodium chloride 5.0g/L, Di-Sodium phosphate 2.5g/L, pH 7.4 + -0.2), and the medium is allowed to stand at 37 ℃ under the most suitable conditions for anaerobic culture.

(1) The culture medium for culturing the streptomyces variabilis is Brain Heart Infusion (Brain Heart Infusion) culture medium (brand OXOID, cat # CM1135), and the main components of the culture medium are Brain Infusion solids 12.5g/L, Beef Heart Infusion solids5.0g/L, protein peptone 10.0g/L, Glucose 2.0g/L, Sodium chloride 5.0g/L, Di-Sodium phosphate 2.5g/L and pH 7.4 +/-0.2. If the agar powder is prepared into solid, 15g/L of agar powder is needed to be added. Sterilizing at 115 deg.C for 30min, and cooling.

(2) The culture medium for culturing the mutan streptococci biofilm is a brain heart infusion-sucrose culture medium, namely, sucrose with the final concentration of 1% is added into the brain heart infusion culture medium. Sucrose was prepared as a 20% stock solution and sterilized by filtration through a 0.22 μm sterile filter.

(3) The strain UA159 of the mutant strain is streaked on a plate containing a brain heart infusion agar solid medium by using a sterile inoculating loop, and the plate is placed upside down in an anaerobic incubator at 37 ℃ for culture until a single colony is obvious.

(4) Scraping and taking out the mutans streptococci UA159 by using a sterile inoculating shovel, respectively transferring into test tubes filled with a brain-heart infusion liquid culture medium, standing in an anaerobic incubator at 37 ℃, and culturing until the liquid is turbid.

(5) Detecting absorbance value (OD) of mutans streptococci under 600nm by using ultraviolet-visible spectrophotometer_600nm)。

(6) The compounds prepared in examples 1 to 24 were precisely weighed on an analytical balance, dissolved by adding DMSO, and then sterilized by filtration through a sterile filter having a pore size of 0.22. mu.m to prepare a stock solution having a final concentration of 1024mg/L, which was stored at-20 ℃ until use.

2. Experimental methods

(1) Culturing for preparing variable chain bacteria UA159 (OD) in logarithmic phase_600nm0.8-1.0) to a final concentration of 5 × 10⁵CFU/mL is ready for use.

(2) And (3) detecting the minimum inhibitory concentration of the compound on the mutagen UA159 by adopting a broth microdilution method. Namely, solutions of compounds (compounds of examples 1 to 24) at different concentrations after dilution in multiple ratios were added to sterile 96-well plates, and the test group with the drug solution was added to the 1 st to 11 th wells, the growth control group was not added to the 12 th well, and the final concentration of the solution of the Streptococcus mutans in each well was 5X 10⁵CFU/mL. The final concentrations of the drugs in the 1 st to 12 th wells were 256, 128, 64, 32, 16, 8, 4, 2, 1, 0.5, 0.25 and 0 μ g/mL, respectively, and the lowest concentration at which bacterial growth was completely inhibited in the wells was set as the Minimum Inhibitory Concentration (MIC), and the experimental results are shown in table 1.

(3) After the bacterial liquid in the small holes is evenly coated on a brain-heart infusion agar solid culture medium, the culture is inverted in an anaerobic incubator at 37 ℃ for 24 hours, the Minimum Bactericidal Concentration (MBC) is positioned according to the minimum concentration without bacterial production, the experiment is effectively repeated for 3 times, and the experimental results are shown in Table 2.

Other strains were tested as above, and the test results are shown in Table 1. The description of the relevant strains and abbreviations is as follows:

TABLE 1 MIC values of compounds for selected bacteria results (units. mu.g/mL)

As can be seen from table 1, the compounds of the present invention have a broad spectrum of antibacterial activity. Meanwhile, in order to test the effect of the compound on oral normal bacteria, an oral normal strain L.delbrueckii is selected for carrying out an experiment, and the result shows that: all the compounds of the invention have little influence on normal oral bacteria, which shows that the compounds of the invention have little influence on the ecological balance of oral flora and have selectivity on cariogenic bacteria. In particular, the compounds II-4, II-6, III-3 and III-5 have particularly obvious antibacterial effect on oral cariogenic bacteria (Table 2).

TABLE 2 MBC value results (in. mu.g/mL) for compounds with outstanding antibacterial activity against mutans bacteria UA159

MIC: a minimum inhibitory concentration; MBC: minimum germicidal concentration

As can be seen from Table 2, compounds II-4 and III-3 have very good killing activity on the planktonic cells of mutans bacteria UA 159.

Example 27: effect of Compounds of the invention on the formation of a biofilm of mutans bacteria

(1) A strain solution of the bacterium of the genus Streptomyces and the compounds of examples 1 to 24 were prepared in the same manner as described in example 25, and the bacterium of the genus Streptomyces in the logarithmic phase was diluted to a final concentration of 1X 10 in a brain heart infusion-sucrose medium⁷CFU/mL is ready for use.

(2) To a sterile 96-well plate, 100. mu.L of the bacterial solution of (1) was added, and wells to which the compound was added were used as an experimental group, and wells to which the compound was not added were used as a control group. Placing the mixture in an anaerobic incubator at 37 ℃ for standing culture for 24 hours.

(3) The planktonic cells in each well were removed and the unadsorbed cells were rinsed with copious amounts of water. To each well was added 200. mu.L of a 0.1% crystal violet solution, and the staining was performed by standing at room temperature for 5min, after which the crystal violet solution was removed and unadsorbed crystal violet was removed by washing with a large amount of water.

(4) 200 mu L of 33% acetic acid solution is added into each hole to dissolve the adsorbed crystal violet, then an enzyme-linked immunosorbent assay is used for detecting the absorbance value under 590nm, the above experiment is effectively repeated for 3 times, and the experiment results are shown in Table 3.

TABLE 3 Effect of Compounds on the formation of a mutans biofilm (units. mu.g/mL)

MBIC: minimum biofilm inhibiting concentration

As can be seen from Table 3, the compounds II-2, II-4, III-2 and III-3 effectively inhibited the formation of a mutans biofilm, and their MBIC values were 4. mu.g/mL, 2. mu.g/mL and 4. mu.g/mL, respectively.

Example 28: experiment of effect of compound on removing mature biofilm of streptomyces variabilis

(1) The procedure described in example 25 was followed to prepare a strain solution of Streptococcus mutans and the compounds of examples 1-24, and the strain in log phase was diluted to a final concentration of 1X 10 in brain heart infusion-sucrose medium⁷CFU/mL is ready for use.

(2) 100. mu.L of the bacterial solution of (1) was added to a sterile 96-well plate, and anaerobic culture was carried out at 37 ℃ for 24 hours to allow the Streptococcus mutans to form a biofilm.

(3) And after the biofilm is formed, discarding the supernatant, adding 150-200 mu L of 0.85% NaCl, standing for 2-3 min, discarding the waste liquid, and repeating for 3 times.

(4) After the waste liquid is discarded, 100uL of fresh BHI culture medium is sequentially added from the second hole to the last hole, 200 muL (256 mug/mL) of compound with the corresponding concentration is added into the first hole, after the first hole is uniformly mixed, 100 muL of compound is taken from the first hole and added into the second hole, the compound is sequentially added into the penultimate hole, the last hole is not added, and the mixture is cultured in an anaerobic incubator at 37 ℃ for 24 hours.

(5) And (4) repeating the operation step (3).

(6) Adding 90 mu L of fresh BHI culture medium into each hole, adding 10 mu L of MTT into each hole to prepare the final concentration of 5mg/mL, wrapping the mixture by using tinfoil paper, and incubating the mixture for 2-4 hours at 37 ℃ in a dark place.

(7) Discarding the supernatant, washing with 0.85% NaCl for 3 times, adding 200. mu.L DMSO, incubating at 37 ℃ in an incubator for 10-15 min, adding 50. mu.L DMSO, measuring the absorbance at 490nm with a microplate reader, and repeating the above experiment effectively for 3 times, wherein the experiment results are shown in Table 4.

TABLE 4 Effect of Compounds on mature biofilms of mutans bacteria (units. mu.g/mL)

Compound (I)	MBEC50	Compound (I)	MBEC50
				I-4	16	II-1	64
II-2	16	III-7	64
				III-4	16	II-4	256
I-3	32	II-5	256
				I-6	32	II-6	256
II-7	32	III-9	256
				III-2	32	III-10	256
III-3	32

MBEC₅₀: minimum biofilm removal concentration

As can be seen from Table 4, compounds II-2, III-4 and I-4 significantly reduced the growth viability of mature biofilms of Streptomyces variabilis, their MBEC₅₀The values were all 16. mu.g/mL.

Example 29: experiment for inhibiting acid production capability of cariogenic virulence factor-mutan bacteria by using compound

(1) A solution of the mutant strain and the compound II-2 were prepared as described in example 25, and the mutant strain in the logarithmic phase was diluted to a final concentration of 5X 10 in a brain heart infusion-sucrose medium⁵CFU/mL is ready for use.

(2) According to the MIC determination of the compound II-2 for mutans, 2 concentrations (1 and 1/2MIC) below the MIC value were added, and the control group was medium without II-2, each in 3 replicates.

(3) Placing in an anaerobic incubator at 37 ℃ for standing culture for 0, 6, 12 and 24 hours, sucking supernatant, monitoring the decline by adopting a precise pH meter, and taking an average value of results, wherein the experimental result is shown in figure 1.

As can be seen from FIG. 1, compound II-2 has a significant inhibitory effect on acid production by Streptomyces variabilis at MIC (4. mu.g/mL) and 1/2MIC (2. mu.g/mL) compared to the control without compound.

Claims (115)

1. A compound having the structure of formula (V):

the A is selected from:

the nitrogen atom moiety in group A is attached to X, and the compound of formula (V) has the following structure:

when the compound of formula (V) is of formula (I)

When being structured, R¹Is substituted phenyl, wherein phenyl may be substituted by fluoro, chloro, bromo, nitro, methoxy, methyl;

when the compound of formula (V) is of formula (II)

When in structure (I), m is 1 or 2, R²Is substituted or unsubstituted phenyl, cyclohexyl, Y is O or S, wherein the phenyl can be mono-substituted or di-substituted by fluorine, chlorine or bromine;

when the compound of formula (V) is of formula (III)

When in structure (I), m is 1 or 2, R¹Is unsubstituted or substituted phenyl, wherein the phenyl group can be substituted by chlorine, bromine, nitro, methyl;

and, the compound does not include

2. A compound according to claim 1, wherein in formula (I), R is¹When the phenyl is substituted, the position of the substituent is selected from C-3 and C-4.

3. A compound according to claim 1, wherein in formula (II), R²In the case of substituted phenyl, the position of said substituentOne or more selected from C-2, C-3 and C-4.

4. The compound of claim 1, wherein in formula (III), R¹When the phenyl is substituted, the position of the substituent is selected from C-3 and C-4.

5. The compound according to any one of claims 1 to 4, wherein the compound is:

1- ((3-bromophenyl) sulfonyl) -N- (5-nitrothiazol-2-yl) piperidine-4-carboxamide, compound I-1;

1- ((4-methoxyphenyl) sulfonyl) -N- (5-nitrothiazol-2-yl) piperidine-4-carboxamide, compound I-2;

1- ((4-chlorophenyl) sulfonyl) -N- (5-nitrothiazol-2-yl) piperidine-4-carboxamide, Compound I-3;

1- ((4-fluorophenyl) sulfonyl) -N- (5-nitrothiazol-2-yl) piperidine-4-carboxamide, compound I-4;

n- (5-Nitrothiazol-2-yl) -1-tosylpiperidine-4-carboxamide, Compound I-5;

1- ((4-bromophenyl) sulfonyl) -N- (5-nitrothiazol-2-yl) piperidine-4-carboxamide, compound I-6;

n- (5-nitrothiazol-2-yl) -4- (3-phenylsulfamoyl) butanamide, Compound II-1;

4- (3- (3-chlorophenyl) ureido) -N- (5-nitrothiazol-2-yl) butanamide, compound II-2;

4- (3-cyclohexylthioureido) -N- (5-nitrothiazol-2-yl) butanamide, compound II-3;

4- (3- (3-chloro-4-fluorophenyl) ureido) -N- (5-nitrothiazol-2-yl) butanamide, compound II-4;

4- (3- (2-bromophenyl) ureido) -N- (5-nitrothiazol-2-yl) butanamide, compound II-5;

4- (3- (4-chlorophenyl) ureido) -N- (5-nitrothiazol-2-yl) butanamide, compound II-6;

5- (3- (3-chloro-4-fluorophenyl) ureido) -N- (5-nitrothiazol-2-yl) pentanamide, compound II-7;

4- ((4-nitrophenyl) sulfonamido) -N- (5-nitrothiazol-2-yl) butanamide, compound III-1;

4- ((4-methylphenyl) sulfonamido) -N- (5-nitrothiazol-2-yl) butanamide, compound III-2;

4- ((4-bromophenyl) sulfonamido) -N- (5-nitrothiazol-2-yl) butanamide, compound III-3;

4- ((4-chlorophenyl) sulfonylamino) -N- (5-nitrothiazol-2-yl) butanamide, compound III-4;

4- ((3-bromophenyl) sulfonamido) -N- (5-nitrothiazol-2-yl) butanamide, compound III-5;

n- (5-nitrothiazol-2-yl) -4- (phenylsulfonylamino) butanamide, Compound III-6;

5- ((4-bromophenyl) sulfonamido) -N- (5-nitrothiazol-2-yl) pentanamide, Compound III-7;

5- ((4-chlorophenyl) sulfonylamino) -N- (5-nitrothiazol-2-yl) pentanamide, Compound III-8;

n- (5-nitrothiazol-2-yl) -5- (phenylsulfonylamino) pentanamide, compound III-9;

5- ((3-bromophenyl) sulfonylamino) -N- (5-nitrothiazol-2-yl) pentanamide, compound III-10.

6. A process for the preparation of a compound of formula (V) as described in claim 1, comprising condensation of 2-amino-5-nitrothiazole starting with HOOC-A-Boc, followed by de-Boc reaction with R¹-SO₂Cl or R²-NCHY reaction to obtain; wherein, A, R¹、R²Y is as defined in claim 1.

7. The method of claim 6, comprising performing the following reaction:

wherein the compound of formula (V) is of the structure:

A、R¹、R²y, m, n are as defined in claim 1.

8. The method of manufacturing according to claim 6, comprising the steps of:

(1) 2-amino-5-nitrothiazole and HOOC-A-Boc are subjected to condensation reaction in a solvent A in the presence of an organic base, a condensing agent and an activating agent to generate an intermediate

(2) Intermediates

In an acid solution, reacting at room temperature to remove Boc protecting group to generate an intermediate

(3) Under the condition of acid-binding agent, the intermediate

Dissolving with solvent B and then reacting with R¹-SO₂Cl or R²-reacting the NCHY at room temperature to obtain a compound of formula (V);

wherein, A, R¹、R²Y is as defined in claim 1.

9. The method according to claim 8, wherein in the step (1), the solvent A is selected from the group consisting of anhydrous N, N-dimethylformamide, tetrahydrofuran, acetonitrile, chloroform, dichloromethane, and 1, 4-dioxane.

10. The method according to claim 8, wherein the solvent A is anhydrous dichloromethane and/or DMF.

11. The method according to claim 8, wherein the solvent is used in an amount of 20 to 25mL per mmol of the 2-amino-5-nitrothiazole in the step (1).

12. The method according to claim 8, wherein in the step (1), the organic base is triethylamine, DBU, NMM, DIPEA.

13. The method according to claim 8, wherein in the step (1), the condensing agent is selected from the group consisting of DCC, DIC, EDCI, HATU, HBTU, HCTU, TBTU, TSTU, TNTU, PyBOP.

14. The method of claim 8, wherein the condensing agent is EDCI and/or HATU.

15. The method according to claim 8, wherein in the step (1), the activator is selected from the group consisting of DMAP, HOBt, 4-PPY, HOAT, HOSu, NHPI, NHNI, and PFPOH.

16. The method according to claim 8, wherein in the step (1), the 2-amino-5-nitrothiazole, the HOOC-A-Boc, the condensing agent, the activating agent and the organic base are used in a molar ratio of 1: 1.2-1.7: 1.2-1.7: 1-2.5: 1 to 3.0.

17. The method according to claim 16, wherein in the step (1), the 2-amino-5-nitrothiazole, the HOOC-a-Boc, the condensing agent, the activating agent and the organic base are used in a molar ratio of 1: 1.2: 1.2: 1-2.2: 1 to 2.5.

18. The method according to claim 8, wherein the reaction temperature in the step (1) is 0 to 60 ℃.

19. The method according to claim 18, wherein the reaction temperature in the step (1) is 30 ℃.

20. The method according to claim 8, wherein in the step (2), the acidic solution is selected from the group consisting of HCl/ethyl acetate solution, HCl/methanol solution, and trifluoroacetic acid.

21. The method according to claim 20, wherein in the step (2), the acidic solution is an HCl/ethyl acetate solution.

22. The method according to claim 8, wherein in the step (2), the acidic solution is used in an amount of each millimole of the intermediate

10-15 mL of an acidic solution is used.

23. The method according to claim 8, wherein in the step (3), the reactant is R¹-SO₂And when Cl is contained, the acid-binding agent is inorganic base, and the inorganic base is selected from potassium carbonate, sodium carbonate or sodium bicarbonate.

24. The method according to claim 8, wherein in the step (3), the reactant is R¹-SO₂When Cl, the solvent B is a solvent/water system selected from dioxane/water, tetrahydrofuran/water, and acetone/water.

25. The method according to claim 24, wherein in the step (3), the volume ratio of the solvent to the water in the solvent/water system is 1: 1 to 3.

26. According toThe process according to claim 24, wherein in the step (3), the solvent B is used in an amount of one millimole per millimole of the intermediate

The amount of the solvent B is 8-12 mL.

27. The production method according to claim 8, wherein, in the step (3), the

Acid-binding agent and R¹-SO₂The molar ratio of Cl is 1: 2-4: 1 to 1.5.

28. The method according to claim 27, wherein, in the step (3), the step (3) is carried out

Acid-binding agent and R¹-SO₂The molar ratio of Cl is 1: 3: 1.2.

29. the method according to claim 8, wherein in the step (3), the reactant is R²-NCHY, the acid-binding agent is organic base selected from triethylamine, DBU, NMM, DIPE.

30. The method of claim 29, wherein the organic base is triethylamine.

31. The method according to claim 8, wherein in the step (3), the reactant is R²-NCHY, intermediate

The mol ratio of the acid binding agent to the acid binding agent is 1: 1-1.5.

32. According to claimThe method according to claim 31, wherein in the step (3), the reactant is R²-NCHY, intermediate

The mol ratio of the acid binding agent to the acid binding agent is 1: 1.2.

33. the method according to claim 8, wherein in the step (3), the reactant is R²-NCHY, the solvent B is selected from anhydrous DMF, tetrahydrofuran, acetonitrile, chloroform, dichloromethane, 1, 4-dioxane.

34. The method according to claim 8, wherein the solvent B is anhydrous tetrahydrofuran.

35. The method according to claim 8, wherein in the step (3), the reactant is R²-NCHY, the solvent is used in the following amount: per millimole of intermediate

The amount of the solvent B is 8-12 mL.

36. The method of claim 8, wherein the intermediate is

Acid-binding agent and R²-molar ratio of NCHY 1: 1-1.5: 1 to 1.5.

37. The method of claim 36, wherein the intermediate is

Acid-binding agent and R²-molar ratio of NCHY 1: 1.2: 1.2.

38. a process for the preparation of a compound as claimed in claim 1, wherein, when the compound of formula (V) has the structure of formula (I), it is prepared by the reaction of:

wherein R is¹As defined in claim 1.

39. The method of manufacturing of claim 38, comprising the steps of:

(1) carrying out condensation reaction on 2-amino-5-nitrothiazole and 1-Boc-4-piperidinecarboxylic acid in a solvent A at 0-60 ℃ in the presence of organic alkali, a condensing agent and an activating agent to generate an intermediate 1-1;

(2) the intermediate 1-1 reacts in an acid solution at room temperature to remove the Boc protecting group to generate an intermediate 1-2;

(3) under the condition of using inorganic base as acid-binding agent, the intermediate 1-2 is dissolved by solvent B and then is reacted with R¹The substituted sulfonyl chlorides were reacted at room temperature to give compounds of formula (I).

40. The method according to claim 39, wherein in step (1), the solvent A is selected from DMF, tetrahydrofuran, acetonitrile, chloroform, dichloromethane and 1, 4-dioxane.

41. The method according to claim 39, wherein in the step (1), the solvent A is anhydrous dichloromethane and/or DMF.

42. The method according to claim 39, wherein the solvent A is used in an amount of 20 to 25mL per mmol of 2-amino-5-nitrothiazole in the step (1).

43. The method according to claim 39, wherein in step (1), the organic base is triethylamine, DBU, NMM, DIPEA.

44. The method according to claim 39, wherein in the step (1), the organic base is triethylamine and/or NMM.

45. The method according to claim 39, wherein in the step (1), the condensing agent is selected from the group consisting of DCC, DIC, EDCI, HATU, HBTU, HCTU, TBTU, TSTU, TNTU, PyBOP.

46. The method according to claim 39, wherein in the step (1), the condensing agent is EDCI and/or HATU.

47. The method according to claim 39, wherein in the step (1), the activator is selected from the group consisting of DMAP, HOBt, 4-PPY, HOAT, HOSu, NHPI, NHNI, and PFPOH.

48. The method according to claim 47, wherein in the step (1), the activating agent is HOBt.

49. The method according to claim 39, wherein in the step (1), the molar ratio of the 2-amino-5-nitrothiazole to the 1-Boc-4-piperidinecarboxylic acid, the condensing agent, the activating agent and the organic base is 1: 1.2-1.7: 1.2-1.7: 1.2-1.7: 1 to 3.

50. The method according to claim 49, wherein in the step (1), the molar ratio of the 2-amino-5-nitrothiazole to the 1-Boc-4-piperidinecarboxylic acid, the condensing agent, the activating agent and the organic base is 1: 1.2: 1.2: 1.2: 1.2.

51. the method according to claim 39, wherein in the step (2), the acidic solution is selected from the group consisting of HCl/ethyl acetate solution, HCl/methanol solution, and trifluoroacetic acid.

52. The method according to claim 51, wherein in the step (2), the acidic solution is an HCl/ethyl acetate solution.

53. The method according to claim 39, wherein in the step (2), the amount of the acidic solution is 10 to 15mL per millimole of 1-1 intermediate.

54. The method according to claim 39, wherein in the step (3), the inorganic base is selected from potassium carbonate, sodium carbonate and sodium hydrogen carbonate.

55. The method according to claim 54, wherein in the step (3), the inorganic base is potassium carbonate.

56. The method according to claim 39, wherein in the step (3), the solvent B is a solvent/water system selected from dioxane/water, tetrahydrofuran/water, and acetone/water.

57. The method according to claim 56, wherein in the step (3), the volume ratio of the solvent to the water in the solvent/water system is 1: 1 to 3.

58. The preparation method according to claim 39, wherein in the step (3), the amount of the solvent B is 8-12 mL per millimole of the intermediate 1-2.

59. The process according to claim 39, wherein in the step (3), the intermediate 1 to 2, the inorganic base, R¹The molar ratio of substituted sulfonyl chlorides was 1: 2-4: 1 to 1.5.

60. The method according to claim 59, wherein in the step (3), the intermediate 1-2, the inorganic base, R¹The molar ratio of substituted sulfonyl chlorides was 1: 3: 1.2.

61. a process for the preparation of a compound as claimed in claim 1, wherein, when the compound of formula (V) has the structure of formula (II), it is prepared by the reaction:

wherein R is²M, Y are as defined in claim 1.

62. The method of manufacturing of claim 61, comprising the steps of:

(1) carrying out condensation reaction on 2-amino-5-nitrothiazole and N-Boc-gamma-aminobutyric acid or N-Boc-5-aminopentanoic acid in a solvent A at 0-60 ℃ in the presence of organic base, a condensing agent and an activating agent to generate an intermediate 2-1;

(2) the intermediate 2-1 is reacted in an acid solution at room temperature to remove a Boc protecting group to generate an intermediate 2-2;

(3) under the condition of organic base, dissolving the intermediate 2-2 with a solvent B, and neutralizing with R²Substituted isocyanates or R²Substituted isothiocyanates are reacted at room temperature to give compounds of formula (II).

63. The method according to claim 62, wherein in step (1), the solvent A is selected from the group consisting of anhydrous DMF, tetrahydrofuran, acetonitrile, chloroform, dichloromethane, and 1, 4-dioxane.

64. The method according to claim 62, wherein in step (1), the solvent A is anhydrous dichloromethane and/or DMF.

65. The method according to claim 62, wherein the solvent A is used in an amount of 20 to 25mL per mmol of 2-amino-5-nitrothiazole in the step (1).

66. The method according to claim 62, wherein in step (1), the organic base is selected from triethylamine, DBU, NMM, DIPEA.

67. The method according to claim 62, wherein in the step (1), the organic base is DIPEA and/or NMM.

68. The method of claim 62, wherein in step (1), the condensing agent is selected from the group consisting of DCC, DIC, EDCI, HATU, HBTU, HCTU, TBTU, TSTU, TNTU, PyBOP.

69. The method according to claim 62, wherein in the step (1), the condensing agent is EDCI and/or HATU.

70. The method according to claim 62, wherein in the step (1), the activator is selected from the group consisting of DMAP, HOBt, 4-PPY, HOAT, HOSu, NHPI, NHNI, and PFPOH.

71. The method according to claim 62, wherein in the step (1), the activating agent is DMAP and/or HOBt.

72. The method according to claim 62, wherein in the step (1), the molar ratio of the 2-amino-5-nitrothiazole or the 5-nitro-2-aminothiazole to the N-Boc- γ -aminobutyric acid or the N-Boc-5-aminopentanoic acid, the condensing agent, the activating agent and the organic base is 1: 1.2-1.7: 1.2-1.7: 1-2.5: 1 to 3.0.

73. The method according to claim 72, wherein in the step (1), the molar ratio of the 2-amino-5-nitrothiazole or the 5-nitro-2-aminothiazole to the N-Boc- γ -aminobutyric acid or the N-Boc-5-aminopentanoic acid, the condensing agent, the activating agent and the organic base is 1: 1.2: 1.2: 2.2: 1.2.

74. the method according to claim 62, wherein in the step (2), the acidic solution is selected from the group consisting of HCl/ethyl acetate solution, HCl/methanol solution, and trifluoroacetic acid.

75. The method according to claim 74, wherein in the step (2), the acidic solution is an HCl/ethyl acetate solution.

76. The method according to claim 62, wherein in the step (2), the amount of the acidic solution is 10 to 15mL per millimole of 2-1 intermediate.

77. The method according to claim 62, wherein in the step (3), the organic base is selected from triethylamine, DBU, NMM, DIPEA.

78. The method according to claim 77, wherein in the step (3), the organic base is triethylamine.

79. The method according to claim 62, wherein in the step (3), the molar ratio of the intermediate 2-2 to the organic base is 1: 1 to 1.5.

80. The method according to claim 79, wherein in the step (3), the molar ratio of the intermediate 2-2 to the organic base is 1: 1.2.

81. the method according to claim 62, wherein in the step (3), the solvent B is selected from the group consisting of anhydrous DMF, tetrahydrofuran, acetonitrile, chloroform, dichloromethane and 1, 4-dioxane.

82. The method according to claim 62, wherein in the step (3), the solvent B is anhydrous tetrahydrofuran.

83. The preparation method according to claim 62, wherein in the step (3), the amount of the solvent B is 8-12 mL per millimole of the intermediate 2-2.

84. The method according to claim 62, wherein in the step (3), the intermediate 2-2, the organic base, and R are used as the base²Substituted isocyanates or R²The molar ratio of substituted isothiocyanates is 1: 1-1.5: 1 to 1.5.

85. The method according to claim 84, wherein in the step (3), the intermediate 2-2, the organic base, and R are²Substituted isocyanates or R²The molar ratio of substituted isothiocyanates is 1: 1.2: 1.2.

86. a process for the preparation of a compound as claimed in claim 1, wherein, when the compound of formula (V) has the structure of formula (III), it is prepared by the reaction:

wherein R is¹M is as defined in claim 1.

87. The method of claim 86, wherein the method comprises the steps of:

(1) carrying out condensation reaction on 2-amino-5-nitrothiazole and N-Boc-gamma-aminobutyric acid or N-Boc-5-aminopentanoic acid in a solvent A at 0-60 ℃ in the presence of organic base, a condensing agent and an activating agent to generate an intermediate 3-1;

(2) the intermediate 3-1 is reacted in an acid solution at room temperature to remove a Boc protecting group to generate an intermediate 3-2;

(3) under the condition of taking inorganic base as an acid-binding agent, dissolving the intermediate 3-2 by using a solvent B, and then neutralizing with R¹The substituted sulfonyl chlorides were reacted at room temperature to give compounds of formula (III).

88. The method according to claim 87, wherein in step (1), the solvent A is selected from the group consisting of anhydrous DMF, tetrahydrofuran, acetonitrile, chloroform, dichloromethane, and 1, 4-dioxane.

89. The method according to claim 87, wherein in step (1), the solvent A is anhydrous dichloromethane and/or DMF.

90. The method according to claim 87, wherein the solvent A is used in an amount of 20 to 25mL per mmol of 2-amino-5-nitrothiazole in the step (1).

91. The method according to claim 87, wherein in step (1), the organic base is triethylamine, DBU, NMM, DIPEA.

92. The method according to claim 87, wherein in step (1), the organic base is DIPEA and/or NMM.

93. The method of claim 87, wherein in step (1), the condensing agent is selected from the group consisting of DCC, DIC, EDCI, HATU, HBTU, HCTU, TBTU, TSTU, TNTU, PyBOP.

94. The method of claim 87, wherein in step (1), the condensing agent is EDCI and/or HATU.

95. The method according to claim 87, wherein in the step (1), the activator is selected from the group consisting of DMAP, HOBt, 4-PPY, HOAT, HOSu, NHPI, NHNI, and PFPOH.

96. The method of claim 87, wherein in step (1), the activating agent is DMAP and/or HOBt.

97. The method according to claim 87, wherein in the step (1), the molar ratio of the 2-amino-5-nitrothiazole to the N-Boc- γ -aminobutyric acid or N- (t-butoxycarbonyl) -5-aminopentanoic acid, the condensing agent, the activating agent, and the organic base is 1: 1.2-1.7: 1.2-1.7: 1.0-2.5: 1.0 to 3.0.

98. The process according to claim 97, wherein in the step (1), the molar ratio of the 2-amino-5-nitrothiazole to N-Boc- γ -aminobutyric acid or N- (t-butoxycarbonyl) -5-aminopentanoic acid, the condensing agent, the activating agent, and the organic base is 1: 1.2: 1.2: 2.2: 1.2.

99. the method according to claim 87, wherein in the step (2), the acidic solution is selected from the group consisting of HCl/ethyl acetate solution, HCl/methanol solution, and trifluoroacetic acid.

100. The method according to claim 99, wherein in the step (2), the acidic solution is an HCl/ethyl acetate solution.

101. The method according to claim 87, wherein in the step (2), the amount of the acidic solution is 10-15 mL per millimole of 3-1 intermediate.

102. The method according to claim 87, wherein in the step (3), the inorganic base is selected from potassium carbonate, sodium carbonate or sodium bicarbonate.

103. The method according to claim 102, wherein in the step (3), the inorganic base is potassium carbonate.

104. The method according to claim 87, wherein in the step (3), the solvent B is a solvent/water system selected from dioxane/water, tetrahydrofuran/water and acetone/water.

105. The method according to claim 104, wherein in the step (3), the volume ratio of the solvent to the water in the solvent/water system is 1: 1 to 3.

106. The preparation method according to claim 87, wherein in the step (3), the amount of the solvent B is 8-12 mL per millimole of the intermediate 3-2.

107. The method according to claim 87, wherein in the step (3), the intermediate 3-2, the inorganic base, and R are selected from the group consisting of¹The molar ratio of substituted sulfonyl chlorides was 1: 2-4: 1 to 1.5.

108. The method according to claim 87, wherein in the step (3), the intermediate 3-2, the inorganic base, and R are selected from the group consisting of¹The molar ratio of substituted sulfonyl chlorides was 1: 3: 1.2.

109. a composition comprising a compound of any one of claims 1 to 5, or a pharmaceutically acceptable salt thereof.

110. The composition according to claim 109, wherein the composition comprises a pharmaceutically acceptable carrier, adjuvant and/or vehicle.

111. A pharmaceutical formulation or cleaning product comprising a compound according to any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier, adjuvant and/or vehicle.

112. The pharmaceutical preparation or cleaning article of claim 111 wherein the pharmaceutical preparation is a solid preparation, a topical preparation, a spray, or a liquid preparation.

113. The pharmaceutical formulation or cleaning product of claim 111, wherein the cleaning product is a toothpaste, mouthwash, or disinfectant.

114. Use of a compound according to any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof or a pharmaceutical composition comprising the same as an active ingredient for the manufacture of a medicament or cleaning product for inhibiting and/or killing planktonic cells of oral bacteria; and/or the presence of a gas in the gas,

the application of the compound of the formula (I) or the pharmaceutically acceptable salt thereof or the pharmaceutical composition taking the compound as the active ingredient in preparing the medicines or cleaning products for inhibiting the biofilm formation caused by the streptomyces mutans or in preparing the medicines or cleaning products for removing the biofilm caused by the streptomyces mutans.

115. Use of a compound according to any one of claims 1 to 5, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing the same as an active ingredient, for the manufacture of an oral mutans biofilm inhibitor, or for the manufacture of a pharmaceutical formulation against caries.

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