HK1199451B - Substituted quinary aza heterocyclic salt compound and its uses in therapeutic protein aging-related disease - Google Patents

Description

Substituted five-membered azacyclo salt compound and application thereof in treating protein aging related diseases

The divisional application is based on the original Chinese patent application with the application number of 200610002391.6, the application date of 27.1.2006 and the name of 'a new substituted pentanitrogen heterocyclic salt compound and the application thereof in treating diseases related to protein aging'.

Technical Field

The present invention relates to substituted five-membered azacyclic salt compounds, processes for their preparation, pharmaceutical compositions containing them and the use of said compounds in the prevention or treatment of diseases or conditions associated with AGEs, including (i) increasing skin elasticity or reducing skin wrinkles, (ii) treating diabetes, (iii) treating or alleviating the sequelae of diabetes, (iv) treating or alleviating kidney damage, (v) treating or alleviating vascular damage, (vi) treating or alleviating hypertension, (vii) treating or alleviating retinopathy, (viii) treating or alleviating lens protein damage, (ix) treating or alleviating cataracts, (x) treating or alleviating peripheral neuropathy, and (xi) treating or alleviating osteoarthritis.

Background

Reactions between sugars and proteins are known to occur. As early as 1912, Maillard found that glucose and other reducing sugars reacted with amino acids, undergoing a series of dehydro-rearrangements to form stable brown pigments. Further studies have found that storing and heating food also produces such pigments formed from sugars and polypeptides. The formation of such pigments reduces the biological activity of the protein and reference is made to us08/588249 for a related application. This non-enzymatic reaction of reducing sugars with free amino acids forms a stable diketo by-product known as the Amadori product. In particular, the beta side chain residue of the amino acid on the surface of heme reacts with glucose to produce heme A1 c. Such reactions also occur with other proteins in the body, such as lens, collagen and neuroproteins (Chemistry, biology and immunology for diabetes and Aging, Advances in Pharmacology, Vol.23, pp.1-34A transdermal Press 1992).

The above reaction is accelerated in the case of diabetic blood glucose levels, and also occurs in the case of normal blood glucose conditions. At the same time, the aging process is closely related to the formation of lipofuscin. Also collagen aging can be mimicked in vitro with sugars and collagen. The glucose-induced collagen products are captured by other proteins and react, thus causing a cross-linking reaction between the proteins. This glucose-induced cross-linking reaction produces Advanced Glycosylation Endproducts (AGEs). AGEs are known to be associated with complications of diabetes, and the normal aging process also causes an increase in AGEs. AGEs in vivo are not only due to their abnormal pathological chemical structure, but also because they are recognized by some specific receptors, causing complex diabetes and age-related pathological changes.

Currently, there are several treatments that prevent the accumulation of AGEs. One such approach is described in USP4758583, wherein the lead aminoguanidine and its analogs prevent the formation of AGEs by reacting with early glycosylation products thereby preventing further conversion of glycosylation products to AGEs and also preventing further crosslinking of AGEs with tissue. The effectiveness of this method was evaluated in animal models of diabetic and aged rats, as well as other indicators of such conditions as macrovascular, renal and neuropathological events. These data were summarized by Vlassara et al. (Vlassara et al, 1994 Biology of Diseases, "Pathology effects of Advanced glycation: Biochemical, biological and clinical informatics for diabetes and aging" Laboratory Investigation 70:138-

Another way to control AGEs in tissues, particularly AGEs cross-linked products that have formed and accumulated in tissues, which cross-linked products lead to clinical or subclinical pathological changes, is to reverse or cleave already formed AGEs cross-linked products. This method of cleaving AGEs has been shown to be effective by Vassan et al (Vassan et al, Nature.1996, Vol.382(18) 275-. The compounds, formulations and methods disclosed in U.S. Pat. No. 4, 5656261 and U.S. Pat. Nos. 08/588249 and 08/848776 all cleave the already formed AGEs cross-linked structures in vivo and in vitro. Studies have shown that these compounds have a good effect on cardiovascular diseases caused by aging (Wolffenbuttel et al, 1998, "openers of advanced Glycation End Products research in vitro properties", Proc. Nat. Acad. Sci. U.S. A.95: 4630-4634). In these studies, AGEs lytic agents administered to diabetic rats for 9 weeks reversed the arteriosclerosis due to diabetes for 1-3 weeks. The parameters improved were cardiac output, peripheral resistance, body artery compliance, aortic input resistance, and carotid compliance (USP 6319934).

Disclosure of Invention

The object of the present invention is to find and develop a small molecule cleavage agent acting on AGEs, which cleaves already formed AGEs to prevent protein cross-linking and cleaves already cross-linked proteins to promote protein metabolism, to treat or prevent various pathological changes caused by the increase of AGEs in vivo, including increasing skin elasticity or reducing skin wrinkles, to treat diabetes or to treat or alleviate sequelae of diabetes, kidney damage, vascular damage, hypertension, retinopathy, lens protein damage, cataracts, peripheral neuropathy or osteoarthritis. Meanwhile, the glycosylated protein acted by the protein crosslinking structure cracking agent is not limited to human protein, but also comprises plant protein or animal protein in crops, so that the application of the protein crosslinking structure cracking agent in the preservation of the plant protein and the animal protein in the crops can be expanded.

The present inventors have found that the compounds of formula I as described below can be used for the treatment and/or prevention of various diseases caused by glycosylation of proteins.

The present inventors have found that the compounds of formula I have better AGEs lytic activity and lower toxicity in various models in vitro and in vivo than the preferred compound ALT-711 disclosed in USP 5656261.

Accordingly, a first aspect of the present invention relates to a compound of formula I, or a pharmaceutically acceptable salt or hydrate thereof,

wherein:

x is O or S, and X is O or S,

y is O or S, and Y is O or S,

q is O or NH, and the compound is,

R₁and R₂May be the same or different and are independently selected from hydrogen, C₁～C₄Alkyl radical, C₂～C₄Alkenyl and hydroxy C₁～C₄Alkyl, or R₁And R₂Are linked to form aromatic ring Ar₂Or a five-or six-membered aliphatic ring,

R₃is hydrogen, C₁～C₈Straight or branched alkyl, C₂～C₈Straight-chain or branched alkenyl radical, C₃～C₈Cycloalkyl, hydroxy, C₁～C₄Alkoxy radical, C₁～C₄An alkylamino group, a nitrile group, or a trifluoromethyl group,

R₄is hydrogen, C₁～C₈Straight or branched alkyl, C₂～C₈Straight-chain or branched alkenyl radical, C₃～C₈A cycloalkyl group, or a monocyclic, bicyclic or tricyclic aromatic carbocyclic or heterocyclic group, wherein each ring consists of 5 to 6 ring atoms, and the heterocyclic group contains 1 to 6 heteroatoms selected from O, S and N; the ring may be unsubstituted or substituted with 1 to 3 substituents selected from the group consisting of: halogen, nitro, hydroxy, hydroxymethyl, trifluoromethyl, trifluoromethoxy, C₁～C₆Straight or branched alkyl, C₂～C₆Straight-chain or branched alkenyl radical, C₁～C₄Alkoxy radical, C₂～C₄Alkenyloxy, phenoxy, benzyloxy, carboxyl and amino groups,

Z^-is a pharmaceutically acceptable acid radical.

According to a preferred embodiment, the present invention relates to a compound of formula I, or a pharmaceutically acceptable salt or hydrate thereof

Wherein:

x is the number of atoms of the group S,

y is O or S, and Y is O or S,

q is O or NH, and the compound is,

R₁and R₂May be the same or different and are independently selected from hydrogen, C₁～C₄Alkyl radical, C₂～C₄Alkenyl and hydroxy C₁～C₄An alkyl group; or R₁And R₂Are linked to form aromatic ring Ar₂Or a five-or six-membered aliphatic ring,

R₃is hydrogen, C₁～C₈Straight or branched alkyl, C₂～C₈Straight-chain or branched alkenyl radical, C₃～C₈Cycloalkyl, hydroxy, C₁～C₄Alkoxy radical, C₁～C₄An alkylamino group, a nitrile group, or a trifluoromethyl group,

R₄is hydrogen, C₁～C₈Straight or branched alkyl, C₂～C₈Straight-chain or branched alkenyl radical, C₃～C₈A cycloalkyl group, or a monocyclic, bicyclic or tricyclic aromatic carbocyclic or heterocyclic group, wherein each ring consists of 5 to 6 ring atoms, and the heterocyclic group contains 1 to 6 heteroatoms selected from O, S and N; the ring may be unsubstituted or substituted with 1 to 3 substituents selected from the group consisting of: halogen, nitro, hydroxy, hydroxymethyl, trifluoromethyl, trifluoromethoxy, C₁～C₆Straight or branched alkyl, C₂～C₆Straight-chain or branched alkenyl radical, C₁～C₄Alkoxy radical, C₂～C₄An alkenyloxy group, a phenoxy group,a benzyloxy group, a carboxyl group and an amino group,

Z^-is a pharmaceutically acceptable acid radical.

According to a more preferred embodiment, the present invention relates to a compound of formula I or a pharmaceutically acceptable salt or hydrate thereof,

wherein:

x is the number of atoms of the group S,

y is a radical of oxygen, Y is O,

q is O, and Q is O,

R₁，R₂，R₃and R₄The definition of (A) is the same as that of (B),

Z^-is a pharmaceutically acceptable acid radical, e.g. F^-、Cl^-、Br^-、I^-Methanesulfonate or p-methylsulfonate, preference being given to hydrobromide and methanesulfonate.

Compounds of the invention include, but are not limited to:

or a pharmaceutically acceptable salt or hydrate thereof.

Among them, 3-benzyloxycarbonylmethyl-4-methyl-thiazole-3-hydrobromide is more preferable.

According to the present invention, pharmaceutically acceptable salts of the compounds of the present invention include inorganic or organic acid salts thereof, including but not limited to: hydrochloride, hydrobromide, hydroiodide, nitrate, sulphate, bisulphate, phosphate, hydrogenphosphate, acetate, propionate, butyrate, oxalate, pivalate, adipate, alginate, lactate, citrate, tartrate, succinate, maleate, fumarate, picrate, aspartate, gluconate, benzoate, methanesulphonate, ethanesulphonate, benzenesulphonate, p-toluenesulphonate and pamoate.

Another aspect of the present invention relates to a process for preparing a compound of formula I, or a pharmaceutically acceptable salt or hydrate thereof, comprising:

a) reacting a thiourea or urea of the formula

With ketones of the formula II

Wherein R is₁And R₂As defined above for the compounds of formula I,

by reaction under halogen catalysis to give compounds of the formula III or by reaction under halogen catalysis according to the literature (J.Amer.chem.Soc.,1949,71,4007),

wherein R is₁，R₂And X is as defined above for compounds of formula I,

b) reacting the compound shown in the formula III with isoamyl nitrite to obtain a compound shown in the formula IV

Wherein R is₁，R₂And X is as defined above, and the process,

c) reacting a compound of formula IV with a compound of formula V

Wherein R is₃Y, Q and R₄Is as defined above for compounds of formula I, X' is a leaving group, such as F, Cl, Br, I, methanesulfonate or p-methanesulfonate,

reacting to obtain the compound of the general formula I

Wherein R is₁，R₂，R₃，X，Y，Q,R₄And Z is as defined above.

Optionally, further hydrolyzing wherein R₄A compound of formula I other than H affords a compound of formula Ia:

wherein R is₁，R₂，R₃X, Y, Q, and Z are as defined above,

wherein Q is O, optionally with C₁～C₈Straight or branched alkyl alcohol, C₂～C₈Linear or branched alkenyl alcohols, C₃～C₈The naphthenic alcohol or aromatic alcohol is subjected to condensation dehydration reaction to obtain various esters.

The resulting compound may be converted from one salt to another salt according to a method known per se, if necessary.

According to a particular embodiment, the compounds of the invention can be prepared by the following reaction scheme:

scheme I:

which comprises reacting a compound of formula IV:

wherein, X is O or S,

R₁and R₂May be the same or different and are independently selected from hydrogen, C₁～C₄Alkyl and C₂～C₄An alkenyl group; or R₁And R₂Are linked to form aromatic ring Ar₂，

With compounds of formula Va

Wherein

Y is O or S, and Y is O or S,

q is O or NH

R₃Is hydrogen, C₁～C₈Straight or branched alkyl, C₂～C₈Straight-chain or branched alkenyl radical, C₃～C₈Cycloalkyl, hydroxy, C₁～C₄Alkoxy radical, C₁～C₄An alkylamino group, a nitrile group, or a trifluoromethyl group,

R₄is C₁～C₈Straight or branched alkyl, C₂～C₈Straight-chain or branched alkenyl radical, C₃～C₈Cycloalkyl, or monocyclic, bicyclic or tricyclic aromatic carbocyclic or heterocyclic radicalA group wherein each ring consists of 5 to 6 ring atoms, and the heterocyclic group contains 1 to 6 heteroatoms selected from O, S and N; the ring may be unsubstituted or substituted with 1 to 3 substituents selected from the group consisting of: halogen, nitro, hydroxy, hydroxymethyl, trifluoromethyl, trifluoromethoxy, C₁～C₆Straight or branched alkyl, C₂～C₆Straight-chain or branched alkenyl radical, C₁～C₄Alkoxy radical, C₂～C₄Alkenyloxy, phenoxy, benzyloxy, carboxyl and amino groups,

to give the compound of the formula I

Wherein X, Y, Q, R₁，R₂，R₃And R₄The definition of (A) is the same as that of (B),

Z^-is Br^-。

In the above reaction scheme, the reaction of the compound of formula IV with the compound of formula Va is carried out in the presence of a solvent such as ethanol, acetonitrile or butanone or in the absence of a solvent when one of the two raw materials is liquid, at 80 ℃ to 100 ℃ for 5 to 96 hours in nitrogen.

The product obtained by the reaction can be kept stand for crystallization and then recrystallized or purified by silica gel column chromatography. The silica gel used in the method can be conventional silica gel for chromatography, the granularity is 10-40 um, and the eluent is prepared from a single or multiple solvents, preferably a mixed solvent prepared from dichloromethane and methanol according to different proportions. After purification, the compounds of formula I according to the invention are obtained.

The reaction conditions and product work-up for the reaction of the compounds of the formula IV with other compounds of the formula V are analogous to those described above.

The compounds of formula IV used in the above schemes are known or can be synthesized using a variety of methods known per se, for example scheme II.

Scheme II:

wherein R is₁、R₂And X is as defined for compounds of formula I.

Wherein the compound of formula II reacts with urea or thiourea under the action of iodine to obtain a compound of formula III, and then the compound of formula IV is obtained by removing amino groups through treatment of isoamyl nitrite in anhydrous tetrahydrofuran. The compound shown in the formula IV can be purified by adopting a high vacuum distillation or silica gel chromatography method, the silica gel used in the method can be silica gel for conventional chromatography, the granularity is 10-40 um, the eluent is prepared by a single solvent or a plurality of solvents, and preferably a mixed solvent prepared by ethyl acetate and cyclohexane according to different proportions.

The compounds of the formula Va used in scheme I are known or can be synthesized using methods known per se. For example, it can be synthesized as follows:

bromination of the α -position of the compound of formula VII with cupric bromide or bromine or NBS gives the compound of formula Va. The purification of the compound of formula Va may be carried out by high vacuum distillation or chromatography. The silica gel used in the method can be conventional silica gel for chromatography, the granularity is 10-40 um, and the eluent is prepared from a single solvent or a plurality of solvents, preferably a mixed solvent prepared from ethyl acetate and cyclohexane according to different proportions. The compound of formula Va must be purified to remove a small amount of the alpha dibromide.

Optionally, wherein R is₄Compounds of formula I other than H may be further hydrolyzed to give compounds of formula Ia:

wherein R is₁，R₂，R₃X, Y, Q, and Z are as defined above,

the hydrolysis may be carried out according to procedures conventional in the art, for example under catalysis by an acid or base.

The compounds of formula Ia in which Q is O may be further combined with various C₁～C₈Straight or branched alkyl alcohol, C₂～C₈Linear or branched alkenyl alcohols, C₃～C₈The naphthenic alcohol or aromatic alcohol is subjected to condensation dehydration reaction to obtain various esters.

When R is₃The resolution of the racemate of the compound related to the invention is preferably a crystallization method with practical value, namely, a chiral resolving agent is added into a solution of the racemate in water, an organic solvent or a mixed solvent formed by the water and the organic solvent to form diastereoisomers, and one of the diastereoisomers is preferentially separated out by utilizing the different solubility of the diastereoisomers in the solvent.

Another aspect of the present invention relates to a pharmaceutical composition comprising at least one compound of formula I, or a pharmaceutically acceptable salt or hydrate thereof, and a pharmaceutically acceptable carrier or excipient. The pharmaceutical composition may be prepared in various forms according to different administration routes. The compounds mentioned in the invention can also be prepared into various pharmaceutically acceptable salts.

The pharmaceutical compositions of the present invention comprise an effective amount of a compound of formula I of the present invention, or a pharmaceutically acceptable salt or hydrate thereof, and one or more suitable pharmaceutically acceptable carriers or excipients. Pharmaceutically acceptable carriers or excipients herein include, but are not limited to: ion exchangers, aluminum oxide, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffer substances such as phosphates, glycerol, sorbic acid, 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, polyvinylpyrrolidone, cellulosic substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, beeswax, lanolin.

A further aspect of the present invention relates to the use of at least one compound of formula I, or a pharmaceutically acceptable salt or hydrate thereof, for the preparation of a medicament for the prevention and/or treatment of various diseases caused by protein glycosylation.

The present invention also relates to a method for preventing and/or treating various diseases caused by protein glycosylation, which comprises administering a prophylactically and/or therapeutically effective amount of at least one compound of formula I, or a pharmaceutically acceptable salt or hydrate thereof, to a patient in need of the prophylaxis and/or treatment.

The compounds of the present invention are potent cross-linked protein cleavage agents, and as compared to ALT-711, the compounds of the present invention have comparable or better capacity to cleave glycosylated proteins and are therefore useful in, but not limited to, (i) increasing skin elasticity or reducing skin wrinkles, (ii) treating diabetes, (iii) treating or alleviating the sequelae of diabetes, (iv) treating or alleviating kidney damage, (v) treating or alleviating vascular damage, (vi) treating or alleviating hypertension, (vii) treating or alleviating retinopathy, (viii) treating or alleviating lens protein damage, (ix) treating or alleviating cataracts, (x) treating or alleviating peripheral neuropathy, (xi) treating or alleviating osteoarthritis.

The glycosylated protein on which the compound disclosed by the invention can act is not limited to human protein, but also comprises plant protein in crops or animal organ protein, so that the compound or the pharmaceutical composition disclosed by the invention can be used for fresh-keeping.

The compounds of the present invention may also be extended to prevent or reverse staining of teeth due to non-enzymatic glycosylation reactions in the oral cavity. The dosage regimen containing the compounds of the present invention may vary depending on the use involved.

Non-enzymatic glycosylation reactions occurring in the oral cavity can lead to staining of teeth. The anti-caries agents currently used accelerate this glycosylation reaction which further leads to staining of the teeth. Recently, a class of cationic antiseptic agents having anti-caries function has been used for conventional oral cleansing. Among these cationic antibacterial agents are alexidine, cetyl pyridinium chlorate, and the like. These agents, in turn, accelerate the Maillard reaction, a critical step in glycosylation reactions, and thus tooth staining (Nordbo, j. dent. res.,58:1429 (1979)). And it has been reported that chlorhexidine and benzalkonium chloride are observed in vitro to catalyze the glycosylation reaction (browning reaction). The addition of chlorhexidine to the mixture of sugar and amino acid accelerates the formation of pigment due to the Maillard reaction.

For the above reasons, the compounds and pharmaceutical compositions thereof according to the present invention can be used in the oral cavity. In particular as an additive in mouth rinses and toothpastes.

In the above uses relating to the compounds of the present invention, suitable forms of non-toxic and pharmaceutically acceptable carriers may be employed in oral cleaning solutions and toothpastes.

Pharmaceutical compositions comprising a compound of the invention may be administered in any of the following ways: oral, aerosol inhalation, rectal, nasal, buccal, topical, parenteral such as subcutaneous, intravenous, intramuscular, intraperitoneal, intrathecal, intraventricular, intrasternal and intracranial injection or infusion, or via an external reservoir. Of these, oral, intraperitoneal or intravenous or topical administration is preferred.

When administered orally, the compounds of the present invention may be formulated in any orally acceptable dosage form, including but not limited to tablets, capsules, aqueous solutions or suspensions. Among these, carriers for tablets generally include lactose and corn starch, and additionally, lubricating agents such as magnesium stearate may be added. Diluents used in capsule formulations generally include lactose and dried corn starch. Aqueous suspension formulations are generally prepared by mixing the active ingredient with suitable emulsifying and suspending agents. If desired, sweetening, flavoring or coloring agents may be added to the above oral dosage forms.

When the compound is used locally, particularly for treating affected surfaces or organs which are easy to reach by local external application, such as eyes, skin or lower intestinal tracts, the compound can be prepared into different forms of local medicinal preparations according to different affected surfaces or organs, and the specific description is as follows:

when administered topically to the eye, the compounds of the present invention may be formulated as a micronized suspension or solution in sterile saline at a pH that is isotonic, with or without the addition of preservatives such as benzylalkenoxides. For ophthalmic use, the compounds may also be formulated in the form of ointments such as vaseline.

When applied topically to the skin, the compounds of the present invention may be formulated in a suitable ointment, lotion, or cream formulation wherein the active ingredient is suspended or dissolved in one or more carriers. Carriers that may be used in the ointment formulation include, but are not limited to, mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyethylene oxide, polypropylene oxide, emulsifying wax and water; carriers that may be used in the lotion or cream include, but are not limited to, mineral oil, sorbitan monostearate, Tween 60, cetyl esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water.

The compounds of the present invention may also be administered in the form of sterile injectable preparations, including sterile injectable aqueous or oleaginous suspensions or solutions. Among the carriers and solvents that may be employed are water, ringer's solution and isotonic sodium chloride solution. In addition, the sterilized fixed oil may also be employed as a solvent or suspending medium, such as a monoglyceride or diglyceride.

It is further noted that the dosage and method of administration of the compounds of the present invention will depend upon a variety of factors including the age, weight, sex, physical condition, nutritional status, activity of the compound employed, time of administration, metabolic rate, severity of the condition, and the subjective judgment of the treating physician. The preferable dosage is 0.01-100 mg/kg body weight/day, wherein the most preferable dosage is in the range of 20-30mg/kg body weight/day.

Examples

The following examples further illustrate the invention without limiting it in any way.

The melting point of the compound was determined by a model SRY-1 melting point apparatus, the temperature being uncorrected.¹H-NMR spectra were determined by a Bruker ARX400 or US Varian Unity Inova600 model nuclear magnetic instrument, FAB mass spectra were determined by a Zabsselect high resolution mass spectrometer.

The general method comprises the following steps: general method for reaction of thiazole mother nucleus and alpha-bromo-ester

On the reaction route, the reaction of the thiazole mother nucleus and various alpha-bromo ester compounds is carried out in the presence of a solvent such as ethanol, acetonitrile or butanone or in the absence of a solvent when one of the two raw materials is liquid, at a temperature of between 80 and 100 ℃ for 5 to 96 hours in nitrogen.

The product obtained by the reaction can be kept stand for crystallization and then recrystallized or purified by silica gel column chromatography. The silica gel used is silica gel for conventional chromatography, the granularity is 10-40 mu m, and the eluent is prepared from a single solvent or a plurality of solvents, such as a mixed solvent prepared from dichloromethane and methanol according to different proportions. And purifying to obtain the target compound.

Example 1: 3-benzyloxycarbonylmethyl-4, 5-dimethyl-thiazole-3-hydrobromide

Following the general procedure described above, using benzyl bromoacetate and 4, 5-dimethylthiazole, the title compound was obtained (0.4g, 26%, oil).

MS[M⁺]＝262.1m/e；¹H-NMR(400MHz,DMSO)2.349(s3H)；2.510(s3H)；5.273(s2H)；5.728(s2H)；7.416-7.427(m5H)；10.104(s1H)。

Example 2: 3-benzyloxycarbonylmethyl-5- (2-hydroxy-ethyl) -4-methyl-thiazole-3-hydrobromide salt

The title compound (0.5g, 28%, oil) was obtained by the general method using benzyl bromoacetate and 4-methyl-5-hydroxyethyl thiazole as starting materials.

MS[M⁺]＝292.1m/e；¹H-NMR(400MHz,DMSO)2.322(s3H)；3.016(t J＝5.2Hz2H)；3.769(t J＝5.2Hz2H)；5.213(s2H)；5.730(s2H)；7.337-7.356(m5H)；10.521(d J＝5.5Hz1H)。

Example 3: 3-benzyloxycarbonylmethyl-4-methyl-thiazole-3-hydrobromide salt

Benzyl bromoacetate and 4-methylthiazole were used according to the general procedure to give the title compound (1.2g, 30%, oil).

MS[M⁺]＝313.9m/e；¹H-NMR(400MHz,CD₃OD)2.512(s3H)；5.293(s2H)；5.600(s2H)；7.356-7.399(m5H)；7.991(s1H)。

Example 4: 3-benzyloxycarbonylmethyl-4, 5,6, 7-tetrahydro-benzothiazole-3-hydrobromide

According to the general procedure, using benzyl bromoacetate and 4,5,6, 7-tetrahydro-benzothiazole,the title compound was obtained (1.3g, 31%, light yellow solid, m.p.160-166 ℃ C.). MS [ M ]⁺]＝288.0m/e；¹H-NMR(400MHz,DMSO)1.798(m4H)；2.681(m2H)；2.903(t J＝4.3Hz2H)；5.269(s2H)；5.702(s2H)；7.312-7.423(m5H)；10.172(s1H)。

Example 5: 3-carboxymethyl-4-methyl-thiazole-3-hydrobromide salt

1g of 3-benzyloxycarbonylmethyl-4-methyl-thiazole-3-hydrobromide salt obtained in example 3 was dissolved in 1N K₂CO₃After completion of the reaction, chloroform 10ml × 3 was extracted, the aqueous layer was taken out, 1N HBr aqueous solution was added dropwise under ice bath to make the solution acidic at pH 2, the aqueous solution was evaporated to dryness, 20 anhydrous ethanol was added to wash the residue, insoluble materials were removed by filtration, and the resulting ethanol solution was evaporated to dryness to give a tan solid, which was the title compound (700mg, yield 50%, m.p.223 ℃ -230 ℃).

MS[M⁺]＝158.2m/e；¹H-NMR(400MHz,DMSO)2.3606(d J＝0.8Hz3H)；5.0936(s2H)；7.7131(d J＝1.7Hz1H)；9.7258(d J＝2.8Hz1H)。

Example 6: ELISA screening test for cleavage of AGE-BSA-collagen cross-linked structures

AGE-BSA and rat tail collagen coated on a 96-well enzyme label plate are crosslinked, and an AGEs crosslinking structure is prepared in vitro. The cleavage of AGEs cross-links by compounds was evaluated by ELISA.

Preparing a tail collagen-coated 96-well enzyme label plate:

normal Wister rats (body weight 200 + -20 g) were sacrificed acutely, the tails removed, and the following tail collagen preparation process was performed at 4 ℃. Firstly, extracting the raw silk of the caudal tendon glue, washing with normal saline and removing non-glue raw silk tissues, then washing with double distilled water for 3 times, shearing, soaking in 0.1% glacial acetic acid at 4 ℃ for 1 week, and shaking at any time in the period. And finally, centrifuging at 8000g for 30min, collecting a centrifuged supernatant collagen solution, and diluting to determine the protein content. Coating a 96-well enzyme label plate (Costar) with 70 mu g of collagen per well, carrying out air drying under aseptic conditions, coating by a preservative film, and storing at 4 ℃ for later use, wherein the Costar is coated on the 96-well enzyme label plate at 4 ℃ for 24 hours.

AGE-BSA preparation:

bovine serum albumin BSA (V) (Roch)50mg/ml and 0.5M glucose were incubated in 0.2MPBS (pH7.4) at 37 ℃ under sterile conditions, protected from light, for 3-4 months, to form glycosylated BSA, i.e., BSA-AGEs. Meanwhile, aglycosylated BSA was prepared with glucose-free BSA. Then, the resulting mixture was dialyzed against 0.01M PBS (pH7.4) to remove unreacted glucose, and BSA-AGEs formation was identified by fluorescence scanning (Exi/Em (395/460nm)) and SDS-PAGE, and protein quantification was carried out by the Lowery method.

The flow of the analysis and determination method comprises the following steps:

coating a 96-well plate with tail collagen, and fully neutralizing acidic collagen for 1h by using pH7.4PBS; SuperBlock (PIERCE) at 37 deg.C, blocking for 1 h; PBST (PBS-Tween) plate 3 times, each oscillation for 1 minutes; AGE-BSA was diluted with PBS to obtain 100. mu.l of AGE-BSA at the maximum crosslinking concentration, which was added to the A, B, C, D row wells of a 96-well plate, BSA at the same concentration was added to the E, F, G, H row wells, PBS in the first 3 rows of 1 as system and reagent blank, and crosslinked with collagen at 37 ℃ for 4 h; PBST washing plate for 4 times, and oscillating for 1min at intervals; diluting a tested compound by adopting pH7.4PBS, adding 100 mu l/hole to 4 holes of AGE-BSA crosslinking and BSA hole respectively, adding 100 mu l/hole of PBS in the same way as a non-lysis control, and incubating for 16h at 37 ℃; PBST washing plate for 4 times, and oscillating for 1min at intervals; adding 80 μ l/well rabbit anti-BSA antibody (1: 500) at 37 deg.C for 50 min; PBST washing plate for 4 times, and oscillating for 1min at intervals; adding 80 μ l/hole horse radish peroxidase labeled goat anti-rabbit IgG (1: 1000) at 37 deg.C for 50 min; PBST plate washing for 3 times, and interval oscillation for 1 min; adding 100 μ l/well of substrate solution TMB (3, 3 ', 5, 5' -tetramethyl benzidine), and stopping light for 20 min; the reaction was stopped with 2mol/L H2SO 4; within 10min, the plate blank wells were zeroed to read OD values at 450nm with a BOBRAD Model550 plate reader.

And (3) data analysis:

the mean OD values were 4-well mean values.

Corrected OD-average of AGE-BSA wells-OD average of BSA wells

The cracking rate is expressed as a percentage of the reduction in OD:

[ (OD mean of PBS well-OD mean of test drug well)/OD mean of PBS well ] ×% ]

According to the above procedure, the test compounds were tested for cleavage at concentrations of 0.1, 0.3, 1mmol/L or less, and the results at concentrations of 0.1 and 1mmol/L are shown in Table 1 (the results are all averages of 3 or more screens):

table 1: ELISA assay for the cleavage rate of Compounds on AGE-BSA-collagen Cross-linking

Claims (13)

1. A compound of the general formula I,

wherein:

x is the number of atoms of the group S,

y is a radical of oxygen, Y is O,

q is O, and Q is O,

R₁is methyl, or with R₂Are connected to form a six-membered fat ring,

R₂is a methyl group, and the compound is,or with R₁Are connected to form a six-membered fat ring,

R₃is a hydrogen atom, and is,

R₄is a benzyl group, and the benzyl group,

Z^-is a pharmaceutically acceptable acid radical.

2. A compound according to claim 1, which is a pharmaceutically acceptable salt thereof,

wherein:

Z^-is F^-、Cl^-、Br^-、I^-Or methanesulfonate, p-toluenesulfonate.

3. A compound selected from:

3-benzyloxycarbonylmethyl-4, 5-dimethyl-thiazole-3-hydrobromide,

3-benzyloxycarbonylmethyl-4-methyl-thiazole-3-hydrobromide,

3-benzyloxycarbonylmethyl-4, 5,6, 7-tetrahydro-benzothiazole-3-hydrobromide,

and a pharmaceutically acceptable salt obtained by replacing a hydrobromide group in the above compound with another pharmaceutically acceptable acid group.

4. A pharmaceutical composition comprising a compound according to any one of claims 1 to 3, and at least one pharmaceutically acceptable carrier.

5. A pharmaceutical composition comprising a compound according to any one of claims 1 to 3, and at least one pharmaceutically acceptable excipient.

6. A process for the preparation of a compound according to any one of claims 1 to 2 and a compound according to claim 3,

3-benzyloxycarbonylmethyl-4, 5-dimethyl-thiazole-3-hydrobromide,

3-benzyloxycarbonylmethyl-4, 5,6, 7-tetrahydro-benzothiazole-3-hydrobromide,

and a pharmaceutically acceptable salt obtained by replacing a hydrobromide group in the above compound with another pharmaceutically acceptable acid group,

the method comprises the following steps:

reacting a compound of formula IV

Wherein R is₁，R₂And X is as defined in claim 1,

with compounds of the formula V

Wherein

R₃Y, Q and R₄As defined in claim 1, X' is a leaving group,

to give the compound of the formula I

Wherein R is₁，R₂，R₃，X，Y，Q,R₄And Z is as defined in claim 1.

7. A process for preparing a compound of formula Ia, said process comprising:

hydrolyzing a compound of formula I according to claim 1 to obtain a compound of formula Ia:

wherein R is₁，R₂，R₃X, Y, Q, and Z are as defined in claim 1.

8. The method of claim 6, wherein X' is F, Cl, Br, I, methanesulfonate, or p-methylbenzenesulfonate.

9. Use of a compound according to any one of claims 1 to 3 in the manufacture of a medicament for the treatment or prevention of a disease or condition associated with advanced glycation end products.

10. Use of a compound according to any one of claims 1 to 3 in the manufacture of a medicament for: (i) increasing skin elasticity or reducing skin wrinkles, (ii) treating diabetes, (iii) treating or ameliorating the sequelae of diabetes, (iv) treating or ameliorating kidney damage, (v) treating or ameliorating vascular damage, (vi) treating or ameliorating hypertension, (vii) treating or ameliorating retinopathy, (viii) treating or ameliorating lens protein damage, (ix) treating or ameliorating cataracts, (x) treating or ameliorating peripheral neuropathy, (xi) treating or ameliorating osteoarthritis.

11. Use of a compound according to any one of claims 1 to 3 for the preparation of a reversal agent for tooth staining or other oral preparations for the prevention and reversal of tooth staining in animals.

12. Use of a compound according to any one of claims 1 to 3 in the manufacture of a medicament for preventing or reversing staining of teeth.

13. Use of a compound according to any one of claims 1 to 3 for the preparation of a vegetable protein or animal protein preservative in crops.