CN101279956B - Matrix metalloprotease inhibitors and synthetic method thereof - Google Patents

Abstract

The invention discloses matrix metalloproteinase inhibitors with general formula(I) and synthesis method thereof. The invention has the advantages that the compounds are all micro-molecular quasi-peptide compounds; the synthesis process is short, the material is easy to get, and the production cost is low.

Description

A kind of matrix metallo-proteinase inhibitor and synthetic method

Technical field

The present invention relates to a kind of matrix metallo-proteinase inhibitor and synthetic method.

Background technology

Along with the aggravation of social senilization, global tumour patient is increasing, and research and development have the social benefit and the remarkable in economical benefits of the antitumor drug of independent intellectual property right.Invasion and attack are the most significant biological characteristics of malignant tumour with transfer, are the major causes that causes cancer patients's clinically dead.Matrix metalloproteinase (MMPs) is that a class can the catalyzed degradation basilar membrane and the important meals proteolytic enzyme of extracellular matrix, and the activity that suppresses this enzyme just can be controlled the generation and the transfer of tumour.MMPs is the endopeptidase family of a zine ion dependent form, all the components that they almost can the degradation of cell epimatrix.Discover that Invasion and Metastasis of malignant cell and tumour cell and matrix thereof induce the level of the MMPs of generation to be proportionate.The catalyst structure domain of MMPs is vital for the MMPs hydrolytic activity, and it has the globular space structure, is made of five strands of β lamellas (four bursts of parallel and one antiparallels) and three long α spirals.Contain two zine ions in the catalysis region, one is structure dependent, and another zine ion then is that catalysis is necessary, and it is positioned at the active centre, and the catalysis zine ion is otherwise known as.But, still do not have ideal MMPs inhibitor now and be applied to clinical since Gross in 1962 and Lapierre in the tadpole body, find the to degrade collagenase (MMP-1) of tail.Therefore, the research and development of this type of medicine are extremely urgent.Three Dimensions Structure according to MMPs, adopt the area of computer aided medicinal design technology of new development in recent years, appropriate design and synthetic ideal MMPs inhibitor, by suppressing the matrix metal proteinase activity of tumor cell surface, reaching the purpose that suppresses tumor cell invasion and transfer, is to find new type anticancer medicine efficient strategy.

Summary of the invention

The purpose of this invention is to provide a kind of matrix metallo-proteinase inhibitor.

Another object of the present invention provides the synthetic method of above-mentioned matrix metallo-proteinase inhibitor.

The present invention is achieved like this, and the chemical general formula of this matrix metallo-proteinase inhibitor is:

R ₂=-H, the straight or branched alkane or the alkene of 1 to 5 carbon atom

R ₃＝-H，-OH，OCH ₃

R ₄＝＝-H，-F，-Cl，-Br.-I，-NO ₂，-CH ₃.-OCH ₃

X＝SO ₂

N=0, the alkane or the alkene of 1 to 3 carbon atom

Or

R ₁＝-OH，

R ₂=-H, the straight or branched alkane or the alkene of 1 complete 5 carbon atoms

R ₃＝-H，-OH，OCH ₃

X＝CO

N=0, the alkane or the alkene of 1 to 3 carbon atom.

Synthetic method step of the present invention is:

(1) at first synthetic benzene sulfonyl-D-L-Ala: the Na that benzene sulfonyl chloride is added the D-L-Ala ₂CO ₃In the solution, under high degree of agitation, 70 ℃ of reaction 0.5hr.Reaction is complete, and the ice bath cooling is transferred pH value to 2.5 with concentrated hydrochloric acid, puts refrigerator overnight, separates out white precipitate, filters, and filter cake is washed with a small amount of frozen water choosing, the distilled water recrystallization, and drying gets object;

(2) synthetic substrate inhibitors of metalloproteinase: benzene sulfonyl-D-L-Ala and N-maloyl imines are dissolved in the dioxane, and room temperature adds the dioxane solution of DCC, room temperature reaction 4hr, reaction is finished, the filtering white precipitate adds 5-phenyl-2-amino-1,3 in the filtrate, the 4-thiadiazoles, room temperature reaction 48hr, the filtering white precipitate, vacuum is melted the filtrate of contracting, excess is dissolved among the EtOAc, and uses saturated Na ₂CO ₃Solution, water, 0.1M HCl and water are respectively given a baby a bath on the third day after its birth time, collect organic phase, remove solvent under reduced pressure, and excess is dissolved among the MeOH, slowly add entry and make and separate out white precipitate, filter, and filter cake is with MeOH-acetonitrile recrystallization, object.

Advantage of the present invention is: designed compound is small molecules and intends peptides, and synthesis route is short, and starting material are easy to get, and production cost is low.

Embodiment

Embodiment:

Synthesizing of benzene sulfonyl-D-L-Ala:

The Na that 2.56ml (20mmol) benzene sulfonyl chloride is added 1.96g (22mmol) D-L-Ala ₂CO ₃Solution (4.45gNa ₂CO ₃Be dissolved in 30ml water) in, under high degree of agitation, 70 ℃ of reaction 0.5hr.Reaction is finished, and the ice bath cooling is transferred pH value to 2.5 with concentrated hydrochloric acid, put refrigerator overnight, separate out white precipitate, filter, filter cake is washed distilled water recrystallization, drying with a small amount of frozen water choosing, get the 1.58g object, productive rate: 34.5%, mp:126-128 ℃, [α]=+ 32.2 ° (0.025g/ml methanol solution, room temperature) (R)-and 5-phenyl-N-((2-benzsulfamide)-1-oxygen-propyl group)-1,3,4-thiadiazoles-2-amine synthetic

1.37g (6mmol) benzene sulfonyl-D-L-Ala and 0.69g (6mmol) N-maloyl imines are dissolved in the dioxane, and room temperature adds the dioxane solution of DCC, room temperature reaction 4hr, reaction is finished, the filtering white precipitate adds 1.0g (5.6mmol) 5-phenyl-2-amino-1,3 in the filtrate, the 4-thiadiazoles, room temperature reaction 48hr, the filtering white precipitate, vacuum is melted the filtrate of contracting, excess is dissolved among the EtOAc, and uses saturated Na ₂CO ₃Solution, water, 0.1M HCl and water are respectively given a baby a bath on the third day after its birth inferior, collect organic phase, remove solvent under reduced pressure, and excess is dissolved among the MeOH, slowly add entry and make and separate out white precipitate, filter, filter cake is with MeOH-acetonitrile recrystallization, get the 0.56g object, productive rate: 25.6%, mp:248-249 ℃

¹H-NMR(d ₆-DMSO)δppm：12.72(s，1H)，8.42-8.40(d，1H)，7.94-7.92(m，2H)，7.82-7.79(dd，2H)，7.56-7.51(m，6H)，4.23-4.16(m，1H)，1.25-1.24(d，1H).MS(ESI)，m/z389(MH ⁺)

The external enzymic activity that presses down

Principle: succinyl gelatin be proved to be can by gelatinase (comprise MMP-2, MMP-9) hydrolysis, the height of the free primary amino concentration that peptide bond hydrolysis produces is proportionate with enzymic activity.Free amine group in the intoxicated protection gelatin of succsinic acid; the primary amine that exposes after the hydrolysis and 2; 4; 6-trinitro-benzene-sulfonic acid (TNBSA) reaction solution; determine amino content by the optical density that detects 450nm wavelength place; thereby determine the activity of gelatinase, reflect the inhibition situation of compound indirectly gelatinase.

Method: the compound 40 μ l that add different concns (1280 μ g/mL, 3200 μ g/mL, 80 μ g/mL, 20 μ g/mL, 5 μ g/mL, 1.25 μ g/mL) in 96 orifice plates respectively, gelatinase solution 10 μ l, succinyl gelatin solution 50 μ l, 50mmol/L sodium borate buffer liquid (pH8.5) is supplied 200 μ l; 100% group does not contain inhibitor, and blank group does not contain gelatinase, all supplies 200 μ l with sodium borate buffer liquid, adds 0.03%TNBS solution 50 μ l again, hatches 60min down for 37 ℃, measures optical density in 450nm wavelength place.Calculate inhibiting rate according to following formula:

Inhibiting rate=(100% optical density, one compound optical density)/(100% optical density, one blank optical density) according to compound concentrations and corresponding inhibition ratio, calculates the IC of each compound ₅₀

Table 1. compound press down enzymic activity

R	IC 50/μM
		-H	373.2
-F	＞1000
		-Cl	96.4
-Br	107.3
		-NO 2	167.8
-CH 3	337.8
		-OCH 3	48.8

Claims (1)

1. the synthetic method of a matrix metallo-proteinase inhibitor, the structural formula of matrix metallo-proteinase inhibitor is:

It is characterized in that synthetic method is:

(1) benzene sulfonyl-D-L-Ala is synthetic:

The Na that benzene sulfonyl chloride is added the D-L-Ala ₂CO ₃In the solution, under high degree of agitation, 70 ℃ of reaction 0.5hr react and finish, and the ice bath cooling with concentrated hydrochloric acid adjust pH to 2.5, is put refrigerator overnight, separates out white precipitate, filters, and filter cake washs with a small amount of frozen water, the distilled water recrystallization, and drying gets benzene sulfonyl-D-L-Ala;

(2) (R)-and 5-phenyl-N-((2-benzsulfamide)-1-oxygen-propyl group)-1,3,4-thiadiazoles-2-amine synthetic:

Benzene sulfonyl-D-L-Ala and N-maloyl imines are dissolved in the dioxane dioxane solution when room temperature adds DCC, room temperature reaction 4hr, reaction is finished, the filtering white precipitate adds 5-phenyl-2-amino-1,3 in the filtrate, the 4-thiadiazoles, room temperature reaction 48hr, filtering white precipitate, vacuum concentrated filtrate, excess is dissolved among the EtOAc, and uses saturated Na ₂CO ₃Solution, water, 0.1M HCl and water are respectively given a baby a bath on the third day after its birth time, collect organic phase, remove solvent under reduced pressure, and excess is dissolved among the MeOH, slowly add entry and make and separate out white precipitate, filter, and filter cake is with MeOH-acetonitrile recrystallization, object.