CN1358720A - Three-site substituted beta-carboline novel compound having anti-HIV and anti-cancer activity - Google Patents

Abstract

Compound of formula (I): wherein: R ₁ represents COR ₄ , CONHR ₄ , wherein R ₄ represents a hydrogen atom, alkyl, aryl, cycloalkyl, heterocycle, mono-(C ₁ -C ₆ ) alkylamino, Mono-(C ₁ -C ₆ ) alkylguanidino, di-(C ₁ -C ₆ ) alkylguanidino, R ₂ represents a hydrogen atom, an alkyl group, COR ₄ , wherein R ₄ is as defined in the description, R ₃ represents a hydrogen atom, an alkyl group or an aryl group, and Ra, Rb, Rc and Rd can be the same or different, and represent groups as defined in the description, and they and their formation with pharmaceutically acceptable acids or bases Addition salts and drugs.

Description

β-Ka Lin new compound with 3-position replacement of anti-HIV and antitumour activity

The present invention relates to the β-Ka Lin new compound that the 3-position replaces.Such new compound has keying action to HIV-1 genome TAR RNA, can suppress combining of HIV-1 Tat albumen and TAR RNA, thereby duplicating of viral interference produces antiviral activity.Suppress the outer experimental result of HIV-1 replisome and show, do not showing under the toxic dosage, such compound exhibits good antiviral activity, the experimental result of this and in-vitro transcription matches.This compounds has antitumor action and anti-OH simultaneously ^*Antioxygenation to dna break.

HIV-1 TAR RNA is a fragment gene group that is present on the HIV-1 virus mRNA, when HIV-1 Tat protein binding during in TAR RNA, has activated transcribing of virus, and transcript is prolonged, and virus is duplicated.The anti-HIV-1 medicine is primarily aimed at virus replication and the key enzyme of transcribing two stages at present: proteolytic enzyme and reversed transcriptive enzyme.But these two kinds of enzymes are because the variation and the genetic heterogeneity and very easily produce resistance of virus itself.Therefore, the investigator gets back in the fundamental research of HIV-1 virus once more, has found the proteic vital role that is combined in the viral life of TAR RNA and Tat.HIV-TAR RNA becomes the focus of Recent study thus.At present, just there are being some research institutions to be engaged in the anti-AIDS drug research that TAR RAN is a target abroad; The domestic still unmanned research of this target being carried out medicinal design except that this group.Therefore,, find a kind of medicine of new anti-HIV-1, so just can reach treatment AIDS patient's purpose as serving as the research target with TAR RNA.

A large amount of 'Beta '-carboline compounds has been described in the document.Especially for its HT _2cThe report of the strong affinity of acceptor.Japan carried out Anticancer Activities to 'Beta '-carboline compound, German Patent GP19502753, claimed 'Beta '-carboline compound with external antagonism cel l proliferation.European patent application EP 373986 claimed 'Beta '-carboline compound Fravopereirine, it has intensive antitumour activity and antiviral (comprising HIV) activity.

Except The compounds of this invention was novel, they also were proved to be anti-HIV-1, anticancer and anti-OH effectively ^*Material to the dna break effect.Therefore they can be used for the antioxidant in acquired immune deficiency syndrome (AIDS), cancer therapy and dietetic food, the disease treatment.

The present invention more properly relates to (I) compound.

Wherein:

R ₁The group of representative is selected from:

COR ₄, R wherein ₄Represent hydrogen atom, straight or branched (C ₁-C ₆) alkyl, aryl, cycloalkyl, heterocycle, list-(C ₁-C ₆) alkylamino, list-(C ₁-C ₆) alkyl guanidine radicals, two-(C ₁-C ₆) the alkyl guanidine radicals, the moieties of each group can be a straight or branched,

CONHR ₄R wherein ₄Be as defined above,

R ₂The group of representative is selected from:

Hydrogen,

Straight or branched (C ₁-C ₆) alkyl,

COR ₄R wherein ₄Be as defined above,

R ₃Represent hydrogen atom, straight or branched (C ₁-C ₆) alkyl or aryl-(C ₁-C ₆) alkyl, the latter's moieties can be a straight or branched,

Ra, Rb, Rc and Rd can be identical or different, and the group of representative is selected from hydrogen, halogen, straight or branched (C independently of one another ₁-C ₆) alkyl, hydroxyl, straight or branched (C ₁-C ₆) alkoxyl group.---wherein each moieties can be a straight or branched;---the alkyl-carbonyl oxygen base of hydroxyl, straight or branched, straight or branched (C ₁-C ₆) acyl group, aryloxy and aryl---(C ₁-C ₆) alkoxyl group;---wherein the alkoxyl group part can be a straight or branched ...

All compounds and at pharmaceutically acceptable acid or the formed additive salt of alkali,

Undoubtedly:

--" aryl " is understood that phenyl, naphthyl, tetralyl, dihydro naphthyl, indenyl or 2, and 3-dihydro indenyl is randomly replaced by one or more identical or different groups separately, and substituting group is selected from halogen, hydroxyl, cyano group, nitro, straight or branched (C ₁-C ₆) alkyl, straight or branched (C ₁-C ₆) alkoxyl group, amino, straight or branched (C ₁-C ₆) alkylamino;--wherein each alkoxyl group part can be a straight or branched--, straight or branched (C ₁-C ₆) acyl group, straight or branched (C ₁-C ₆) alkoxy carbonyl, straight or branched (C ₁-C ₆) alkyl amino-carbonyl and oxo.

--" heterocycle " is understood that saturated or undersaturated list-or two-cyclic group, have aromatics and non-aromatic character, have 5 to 12 annular atomses, contain one, heteroatoms that two or three are identical or different, heteroatoms is selected from oxygen, nitrogen and sulphur, heterocycle is understood that and can be randomly replaced by one or more identical or different substituting groups that substituting group is selected from halogen, hydroxyl, straight or branched (C ₁-C ₆) alkyl, straight or branched (C ₁-C ₆) alkoxyl group, nitro, oxo and amino is (arbitrarily by one or two straight or branched (C ₁-C ₆) the alkyl replacement).

In heterocycle, can be symbolic and do not add any pyridyl, thienyl, furyl, imidazolyl, 4-H-pyrans-4-ketone, pyrazinyl, pyrimidyl, isoxazolyl, tetrazyl, pyrryl, pyrazolyl, quinolyl, isoquinolyl, quinazolyl, pyrrolidyl, piperidyl, piperazinyl, 1 restrictedly mentioned, 2, the 3-thiadiazolyl group ...

In pharmaceutically acceptable acid, can not add any hydrochloric acid, cyanogen bromic acid, sulfuric acid, phosphonic acids, acetate, trifluoroacetic acid, lactic acid, pyruvic acid, propanedioic acid, succsinic acid, pentanedioic acid, fumaric acid, tartrate, toxilic acid, citric acid, xitix, oxalic acid, methylsulfonic acid, dextrocamphoric acid etc. restrictedly mentioned.

In pharmaceutically acceptable alkali, can not add any sodium hydroxide, potassium hydroxide, triethylamine, TERTIARY BUTYL AMINE etc. restrictedly mentioned.

According to a kind of favourable change example, preferably The compounds of this invention is, wherein, and R ₁Be COR ₄R wherein ₄Define suc as formula (I).According to a kind of favourable change example, preferred substituted R ₁Be CONR _4aR _4bWherein work as R _4bWhen representing hydrogen atom, R _4aRepresent strand (C ₁-C ₆) alkylamino, strand (C ₁-C ₆) the alkyl guanidine radicals; R _4aAnd R _4bWhen linking to each other, represent piperazine.

According to the present invention, preferred substituted R ₂And R ₃Be hydrogen atom and single straight chain (C ₁-C ₆) alkyl.Especially advantageously R ₂And R ₃Represent hydrogen atom and methyl.

According to the present invention, preferred compound is:

3-ethylamino-β-Ka Lin-3-methane amide

3-third amino-beta--carboline-3-methane amide

Own amino-beta--the carboline of 3--3-methane amide

9-methyl-3-third amino-beta--carboline-3-methane amide

1-methyl-3-ethylamino-β-Ka Lin-3-methane amide

1-methyl-3-third amino-beta--carboline-3-methane amide

1-methyl-3-piperazinyl-β-Ka Lin-3-methane amide

Own amino-beta--the carboline of 1-methyl-3--3-methane amide

1,9-dimethyl-3-third amino-beta--carboline-3-methane amide

3-(2-guanidine radicals)-ethyl-β-Ka Lin-3-methane amide

3-(3-guanidine radicals)-propyl group-β-Ka Lin-3-methane amide

3-(6-guanidine radicals)-hexyl-β-Ka Lin-3-methane amide

3-(12-guanidine radicals)-dodecyl-β-Ka Lin-3-methane amide

1-methyl-3-(2-guanidine radicals)-ethyl-β-Ka Lin-3-methane amide

1-methyl-3-(3-guanidine radicals)-propyl group-β-Ka Lin-3-methane amide

1-methyl-3-(6-guanidine radicals)-hexyl-β-Ka Lin-3-methane amide

Preferred compound and they and pharmaceutically acceptable acid or the formed additive salt of alkali constitute the part of complete content of the present invention.

The invention still further relates to the preparation method of formula (I) compound, it is characterized in that using (II) compound as raw material:

Wherein Ra, Rb, Rc, Rd and R ₃Be as defined above.

According to conventional one-tenth ester condition in the organic synthesis, obtain formula (III) compound with methyl alcohol.

Wherein Ra, Rb, Rc, Rd and R ₃Be as defined above.

According to the Pictet-Spengler reaction conditions in the organic synthesis, make this formula (III) compound and (IV) compound reaction:

R ₂CHO (IV)

Wherein, R ₂Define suc as formula (I).

Obtain the formula V compound:

Wherein Ra, Rb, Rc, Rd, R ₂And R ₃Be as defined above.

Be subjected to the effect of oxygenant commonly used in the organic synthesis, obtain formula (VI) compound

Wherein Ra, Rb, Rc, Rd, R ₂And R ₃Be as defined above.

According to the amidation condition of routine, use formula (VII) compound treatment:

R _4aNHR _4b (VII)

R wherein _4aAnd R _4bBe as defined above.

Obtain formula (I/a) compound, i.e. a specific examples of formula (I) compound:

Wherein Ra, Rb, Rc, Rd, R ₂, R ₃, R _4aAnd R _4bBe as defined above.

This formula compound is placed under the effect of formula (VIII) compound:

[H ₂N-C(-SCH ₃)＝NH] ₂·H ₂SO ₄ (VIII)

Obtain formula (I/b) compound, i.e. a specific examples of formula (I) compound:

Wherein Ra, Rb, Rc, Rd, R ₂, R ₃And R _4aBe as defined above.

Compound (I/a) and (I/b) constitute the integral body of this compound.If necessary, these compounds carry out purifying according to the purifying process of routine, if necessary, can randomly be converted into additive salt with pharmaceutically acceptable acid or alkali.

Formula (II), (IV), (VII), (VIII) compound are commercial available compounds, or the compound that obtains according to the currently known methods of organic synthesis.

The invention still further relates to pharmaceutical composition, comprise at least a formula (I) compound or with pharmaceutically acceptable acid or the formed additive salt of alkali as activeconstituents, independent or pharmaceutically acceptable, inert, nontoxic vehicle or carrier in conjunction with one or more.

In according to drug regimen of the present invention, can mention especially and be applicable to oral, parenteral (intravenously, muscle or subcutaneous), through skin or transdermal, intranasal, rectum, through tongue, through those of eye or respiratory administration, especially tablet or drageeing, Sublingual tablet, cachet, capsule, lozenge, suppository, creme, ointment, skin gel, can be injected into drinkable preparation, aerosol, eye drops or nasal drop etc.

The compounds of this invention has anti-HIV-1 activity, antitumour activity and anti-OH ^*The anti-oxidant activity of dna breakage.Therefore the drug regimen that contains at least a formula (I) compound can be used for the treatment of HIV-1, the antioxidant in cancer therapy and disease, food, the pharmaceuticals.

As medicine, useful dosage is different because of patient age and body weight, route of administration, disease character and seriousness and any other treatment of being accepted.

The following example is set forth and is limited the present invention absolutely not:

Raw materials used and/or reagent is known product, or according to the product of known operation preparation.

The structure of compound described in embodiment and the synthesis step be according to routine spectroscopic techniques (infrared, NMR, TOF-mass spectrum ...) measure.

Embodiment 1 3-ethylamino-β-Ka Lin-3-methane amide

Steps A: hydrochloric acid tryptophan methyl ester

1.0ml sulfur oxychloride splashes under-4 ℃ in the suspension of 1.0g L-tryptophane and 20ml anhydrous methanol, finishes, and removes ice-water bath, stirring at room is after 6 hours, and 70 ℃-80 ℃ were refluxed 6 hours, and the pressure reducing and steaming solvent obtains expecting product.

Step B:1,2,3,4-tetrahydrochysene-β-Ka Lin-3-methyl-formiate

The product that obtains in the 1.25g steps A is dissolved in the 75ml methyl alcohol, add 37% formaldehyde 0.95ml, after vapor bath refluxed 2 hours, the pressure reducing and steaming solvent, transfer PH8 to 9 with 14% strong aqua, add the 40ml chloroform extraction, use 3 * 20ml chloroform wash water layer again, the combined chloroform layer, anhydrous sodium sulfate drying spends the night, filtering sodium sulfate, the filtrate decompression evaporate to dryness obtains expecting product.

Step C: β-Ka Lin-3-methyl-formiate

The product that obtains among the 0.5g step B is dissolved in the 10ml Glacial acetic acid, in the cooling bath, stirs adding lead tetra-acetate 2.25g down fast, finish, stirred 20 minutes, add 2.5g oxalic acid, stirred 1 hour, and filtered, precipitation is suspended in the mixing solutions of 25ml water and 50ml chloroform, transfer pH to neutral with sodium bicarbonate, it is two-layer to filter branch, collects chloroform layer, wash chloroform layer with saturated common salt, tell chloroform layer, anhydrous sodium sulfate drying spends the night, remove by filter sodium sulfate, the filtrate decompression evaporate to dryness obtains expecting product.

Step D:3-ethylamino-β-Ka Lin-3-methane amide

The product that obtains among the 0.1g step C is dissolved in 4ml chloroform and 5 methyl alcohol, this solution was splashed into the 1ml 1 that is heated to 80 ℃-85 ℃ in 3 hours, in the 2-quadrol, finish, after the reflux 6 hours, concentrating under reduced pressure, the mixed solution of adding 5ml chloroform and 3ml water in the residue fully stirs the back standing over night.Separate out solid next day, filter, wash solid respectively with less water and methyl alcohol, the collection solid is in vacuum-drying, obtains expecting product.

Fusing point: 234-237 ℃

EI-MS：255

¹H-NMR：12.26，9.08，8.92，8.87，8.45，7.69，7.61，3.63，3.03

Embodiment 2:3-third amino-beta--carboline-3-methane amide

Operation uses 1 with embodiment 1 in step D, the 3-propylene diamine is as reactant.

Fusing point: 218-222 ℃

EI-MS：269

¹H-NMR：8.917，8.889，8.835，8.395，7.673，7.591，7.297，3.420，

2.614，1.638

Own amino-beta--the carboline of embodiment 3:3--3-methane amide

Operation uses 1 with embodiment 1 in step D, the 6-hexanediamine is as reactant

Fusing point: 189-193 ℃

EI-MS：311

¹H-NMR：8.93，8.84，8.37，7.67，7.58，7.25，3.42，3.28，2.78，1.82

Embodiment 4:9-methyl-3-third amino-beta--carboline-3-methane amide

Operation uses 1-methyl-L-tryptophane as reactant in steps A with embodiment 2.

Fusing point: 116-118 ℃

EI-MS：282

Embodiment 5:1-methyl-3-ethylamino-β-Ka Lin-3-methane amide

Step e: 1-methyl isophthalic acid, 2,3,4-tetrahydrochysene-β-Ka Lin-3-methyl-formiate

Get 0.5g L-tryptophane, add the mixed solution of 1N sulfuric acid 2.5ml and 8ml water, stir adding 40% acetaldehyde 3.6ml down, 40 ℃ of heating in water bath 30 minutes, stirring at room 2 hours, reflux is 1.5 hours in vapor bath, be chilled to room temperature, with dense ammonia thermal transfer pH to 8, filter, precipitate with cold water washing, collecting precipitation, room temperature is dried, with the precipitation that obtains set by step B carry out esterification, obtain expecting product.

Step F: 1-methyl-β-Ka Lin-3-methyl-formiate

Get the product that the 1.5g step e obtains, add 4.2g sulphur, the 100ml dry xylene refluxed 4.5 hours, reaction solution is chilled to 5 ℃ spends the night, filter next day, with 40ml cold xylene and the washing precipitation successively of lower boiling sherwood oil, collecting precipitation, vacuum-drying obtains the re-set target compound.

Compound D reaction set by step with step F obtains obtains expected compound.

Fusing point: butter shape

EI-MS：268

¹H-NMR：11.15，8.72，8.34，7.64，7.58，6.8，3.1-3,67，2.83

Embodiment 6:1-methyl-3-third amino-beta--carboline-3-methane amide

Operation is with embodiment 5, and with 1, the 3-propylene diamine is as reactant in step D.

Fusing point: 175-177 ℃

EI-MS：292

¹H-NMR：12.03，8.77，8.35，7.67，7.59，7.30，3.56，3.33，2.83，1.9

Own amino-beta--the carboline of embodiment 7:1-methyl-3--3-methane amide

Operation is with embodiment 5, and with 1, the 6-hexanediamine is as reactant in step D

Fusing point: 243 ℃ (carbonization)

EI-MS：324

Embodiment 8:1-methyl-3-piperazinyl-β-Ka Lin-3-methane amide

Operation uses piperazine as reactant in step D with embodiment 5.

Fusing point: 192-194 ℃

EI-MS：294

Embodiment 9:1,9-dimethyl-3-third amino-beta--carboline-3-methane amide

Operation is with embodiment 5, in step e with 1-methyl-L-tryptophane as reactant, in step D with 1,3 propylene diamine as reactant.

Fusing point: 149-151 ℃

EI-MS：296

Embodiment 10:3-(2-guanidine radicals)-ethyl-β-Ka Lin-3-methane amide

Step G: take by weighing S-first isothiourea vitriol 0.021g, add 0.03ml water and 0.03ml 2M sodium hydroxide, drip the compound 0.1g that obtains among the embodiment 1 under stirring fast and be suspended in 2ml H ₂The mixed solution of O finishes, stirring at room two days, and 4 ℃ of placements are spent the night, and filter, and precipitation is used a spot of water and methanol wash respectively, collecting precipitation, vacuum-drying obtains expecting compound.

Fusing point: 312-313 ℃

TOF-MS：297.0486

Embodiment 11 3-(3-guanidine radicals)-propyl group-β-Ka Lin-3-methane amide

Operation is with embodiment 10, and the compound that use embodiment 2 obtains in step G is as reactant.

Fusing point: 199-201 ℃

TOF-MS：311.0407

¹H-NMR：8.92，8.81，8.37，7.64，7.58，7.27，3.41，3.11，1.75

¹³C-NMR：165.08，157.12，141.10，139.61，137.23，132.47，128.38，

128.00，122.07，120.96，119.75，113.73，112.30，38.39，36.23，28.84

Embodiment 13 3-(12-guanidine radicals)-dodecyl-β-Ka Lin-3-methane amide

Operation is with embodiment 10, usefulness in step D, 1,12-diamino-dodecane is as reactant, this product in step G as reactant.

Fusing point: 199-201 ℃

TOF-MS：437.1623

Embodiment 14:1-methyl-3-(2-guanidine radicals)-ethyl-β-Ka Lin-3-methane amide

Operation uses compound that embodiment 5 obtains as reactant in step G with embodiment 10.

Fusing point: 242-245 ℃

TOF-MS：297.0486

Embodiment 15 1-methyl-3-(3-guanidine radicals)-propyl group-β-Ka Lin-3-methane amide

Operation is with embodiment 10, and the compound that use embodiment 6 obtains in step G is as reactant.

Fusing point: 270-272 ℃

TOF-MS：325.0608

¹H-NMR：12.03，8.66，8.28，7.64，7.53，7.23，3.36，3.12，2.82，1.69

¹³C-NMR：165.12，157.13，140.94，140.84，138.99，135.85，128.13，

127.31，122.03，121.42，119.77，112.27，111.99，36.30，29.06，20.41(2C)

Embodiment 16 1-methyl-3-(6-guanidine radicals) hexyl-β-Ka Lin-3-methane amide.

Operation is with embodiment 10, and the compound that use embodiment 7 obtains in step G is as reactant.

Fusing point: 196-198 ℃

TOF-MS：367.0860

The pharmacological research of The compounds of this invention.

Embodiment 17 anticancer shaker tests are tested with KB, Hela, HL-60, BGC and BEL-7402 tumour cell respectively, with mtt assay or srb assay.

Mtt assay: respectively with growth conditions HL-60 good, that be in logarithmic phase, Bel-7402, KB and Hela cell with 1 * 10 ⁴Individual/mL concentration is inoculated in 96 orifice plates, 37 ℃ of 5%CO ₂Cultivated 24 hours in the incubator.Abandon old liquid, renew nutrient solution, add the compound of sterilising treatment, continue to cultivate after 48 hours, discard nutrient solution, every hole adds RPMI 1640 (the containing 10% calf serum) nutrient solution that 20mL contains 5mg/mL MTT, continues to cultivate 4 hours.Centrifugal, 2500rpm, 20min, the sucking-off supernatant, room temperature is dried.Add a certain amount of dmso solution purple residue, on 570nm place microplate reader, measure absorption value.

Srb assay: the same with mtt assay.

Respectively with growth conditions HL-60 good, that be in logarithmic phase, Bel-7402, KB and Hela cell with 1 * 10 ⁴Individual/mL concentration is inoculated in 96 orifice plates, 37 ℃ of 5%CO ₂Cultivated 24 hours in the incubator.Abandon old liquid, renew nutrient solution, add the compound of sterilising treatment, continue to cultivate after 48 hours, get supernatant, add 100 μ l 10%TCA (Tricholroacetic Acid) in each aperture, leave standstill 5 minutes after, in 4 ℃ place 1 hour fixing.Outwell stationary liquid then, each aperture washes with water 5 times.After the drying at room temperature, every hole adds 0.4%SRB100 μ l, and room temperature was placed after 10 minutes, washes 5 times with 1%HOAc (acetic acid), and dry air adds the every hole of 10mMTris 200 μ l/, the vibration dissolving, and the place measures the OD value in the single wavelength of 540nm

Embodiment 18: anxious poison test

Medication preparation is become hydrochloride, standby with physiological saline solution.At first carry out trial test, find out 0% and 100% and estimate lethal quantity; According to this result, set five dosage groups then, the dosage spacing is 1: 0.8.(body weight 20 ± 2g) is divided into five groups to Kunming mouse at random, and 10 every group, male and female half and half are observed animal toxicity response situation and death condition in 14 days continuously after the administration.Add up dead animal number in 14 days, the input computer calculates its LD with Bliss method statistics program ₅₀Value and 95% average fiducial limit.Record compound 3-third amino-beta--carboline-3-methane amide to the medium lethal dose (LD of mouse according to aforesaid method through intraperitoneal administration ₅₀) be 170mg/Kg.

Embodiment 19: the combination of transcriptional level antagonism HIV-1 Tat/TAR RNA

We have successfully made up two plasmids.One is Tat gene expression plasmid (plasmid I), and another is to be promotor with HIV-1 LTR fragment, contains the plasmid (plasmid II) of E.C. 2.3.1.28 (CAT) reporter gene.Behind the transformed into escherichia coli, cut evaluation through enzyme and obtain positive colony.Utilize calcium phosphate to mediate this two kinds of plasmid cotransfection 293 cells, add 3 kinds of samples after 24 hours, concentration is 30 μ mol/L.Collect 48 hours culture supernatant, under the 405/490nm wavelength, detect CAT activity in the cell conditioned medium liquid with CAT ELISA test kit.Investigate sample to the interactional influence of Tat-TAR.

With the plasmid I that does not add sample, the CAT activity of II cotransfection system is 100%, represents the CAT activity of application of sample cotransfection system with relative reactivity.

Embodiment 20: suppress HIV-1 and duplicate in vitro tests.

Adopt MT ₄Cell and HIV IIIB strain are tested, and the virus quantity of use is divided into 100TCID ₅₀And 1000TCID ₅₀(tissue cultured infection dose).MT ₄Medicine under cell and HIV IIIB strain and the various dose was cultivated after 5 days, examined under a microscope CPE (cytopathy---cavity, swelling, fusion etc.).Set up five groups of experiments:

Contrast: medicine+MT ₄Cell

Blank: MT ₄Cell+HIV IIIB

Negative control: water+MT ₄Cell+HIV IIIB

Positive control: AZT+MT ₄Cell+HIV IIIB

Experimental group: medicine+MT ₄Cell+HIV IIIB

Embodiment 21: anti-OH ^*To the anti-oxidant activity of dna break, adopt plasmid pBR322 to test, with the reaction induced OH of Fenton ^*Generation.

This test is widely used in estimating medicine antagonism OH ^*Protective capability to dna break.

Steps A:

With compound (5/0.5mM) 2 μ l and tetraacethyl disodium (3mm), potassium phosphate buffer (50mm, pH7.4), hydrogen peroxide (3mm), ferrous sulfate (1.6mm) and each 2ul of pBR322 plasmid DNA, be mixed in the 1.5ml Eppendorf pipe, the ratio that guarantees ferrous sulfate/tetraacethyl disodium is 0.53, and cumulative volume is 12ul (insufficient water replenishes), 37 ℃ hatch 30 minutes after, every pipe adds 2ul bromine Fen indigo plant, agarose gel electrophoresis (0.8% agar powder, 1XTAE, pH7.4,600mv, about 1 hour) observations under the ultraviolet transilluminator.

Blank: plasmid DNA

Negative control: do not add any anti-oxidation active substance

Positive control: V _E(5mM)

Claims (21)

1. The compound of formula (I): wherein: The group represented by R ₁ is selected from: COR ₄ , wherein R ₄ represents a hydrogen atom, linear or branched (C ₁ -C ₆ ) alkyl, aryl, cycloalkyl, heterocyclic, mono-(C ₁ -C ₆ )alkylamino, mono-(C ₁ -C ₆ )alkylguanidino, di-(C ₁ -C ₆ )alkylguanidino, the alkyl part of each group can be straight or branched of. CONHR ₄ wherein R ₄ is as defined above. The group represented by _R is selected from: hydrogen, Straight chain or branched (C ₁ -C ₆ ) alkyl, R ₄ in COR ₄ is as defined above, R ₃ represents a hydrogen atom, a linear or branched (C ₁ -C ₆ )alkyl group or an aryl-(C ₁ -C ₆ )alkyl group, and the alkyl portion of the latter may be linear or branched. Ra, Rb, Rc and Rd can be the same or different, and the groups independently represented by each other are selected from hydrogen, halogen, linear or branched (C ₁ -C ₆ ) alkyl, hydroxyl, linear or branched (C ₁ -C ₆ )alkoxy. - where each alkyl moiety may be straight-chain or branched; - hydroxyl, straight-chain or branched alkylcarbonyloxy, straight-chain or branched (C ₁ -C ₆ ) acyl, aryloxy and Aryl - (C ₁ -C ₆ )alkoxy; - wherein the alkoxy moiety may be straight chain or branched... . All compounds and their pharmaceutically acceptable addition salts with acids or bases, no doubt: - "aryl" is understood as phenyl, naphthyl, tetrahydronaphthyl, dihydronaphthyl, indenyl or 2,3-dihydroindenyl, each optionally replaced by one or more identical or different radicals Group substitution, the substituent is selected from halogen, hydroxyl, cyano, nitro, straight or branched (C ₁ -C ₆ ) alkyl, straight or branched (C ₁ -C ₆ ) alkoxy, amino, Straight-chain or branched (C ₁ -C ₆ )alkylamino;—wherein each alkoxy moiety may be straight-chain or branched—, straight-chain or branched (C ₁ -C ₆ )acyl, straight-chain Chain or branched (C ₁ -C ₆ )alkoxycarbonyl, straight or branched (C ₁ -C ₆ )alkylaminocarbonyl and oxo. - "heterocycle" is understood as a saturated or unsaturated mono- or bi-cyclic group, of aromatic and non-aromatic character, having from 5 to 12 ring atoms, containing one, two or three identical or different Heteroatoms, heteroatoms are selected from oxygen, nitrogen and sulfur, heterocycles are understood to be optionally substituted by one or more identical or different substituents selected from halogen, hydroxyl, straight or branched (C ₁ -C ₆ ) alkyl, straight or branched (C ₁ -C ₆ ) alkoxy, nitro, oxo and amino (optionally replaced by one or two straight or branched (C ₁ -C ₆ ) Alkyl substitution. In heterocycles, pyridyl, thienyl, furyl, imidazolyl, 4-H-pyran-4-one, pyrazinyl, pyrimidinyl, Isoxazolyl, tetrazolyl, pyrrolyl, pyrazolyl, quinolinyl, isoquinolyl, quinazolinyl, pyrrolidinyl, piperidinyl, piperazinyl, 1,2,3-thiadi Azolyl.... 2. Compounds of formula (I) according to claim 1, characterized in that R ₁ represents a COR ₄ radical, wherein R ₄ is as defined for formula (I). They and their addition salts with pharmaceutically acceptable acids or bases. 3. The compound of formula (I) according to claim 1 or 2, characterized in that R ₁ represents CONHR _4a , R _4a represents single chain (C ₁ -C ₆ ) alkylamino, single chain (C ₁ -C ₆ ) alkane Guanidino. They and their addition salts with pharmaceutically acceptable acids or bases. 4. The compound of formula (I) according to claim 1, characterized in that the group represented by R ₂ is selected from: hydrogen, straight or branched chain (C ₁ -C ₆ ) alkyl, COR ₄ , wherein R ₄ is as defined above. They and their addition salts with pharmaceutically acceptable acids or bases. 5. Compounds of formula (I) according to claim 1 or 4, characterized in that _R2 represents hydrogen or methyl. They and their addition salts with pharmaceutically acceptable acids or bases. 6. Compounds of formula (I) according to claim 1, characterized in that R ₃ represents a hydrogen atom, a linear or branched (C ₁ -C ₆ )alkyl group or an aryl (C ₁ -C ₆ )alkyl group, The latter alkyl moiety may be linear or branched. They and their addition salts with pharmaceutically acceptable acids or bases. 7. Compounds of formula (I) according to claim 1 or 6, characterized in that _R3 represents a hydrogen atom or a methyl group. They and their addition salts with pharmaceutically acceptable acids or bases. 8. The compound of formula (I) according to claim 1, which is 3-propylamino-β-carboline-3-carboxamide. It and its addition salts with pharmaceutically acceptable acids or bases. 9. The compound of formula (I) according to claim 1, which is 1-methyl-3-ethylamino-β-carboline-3-carboxamide. It and its addition salts with pharmaceutically acceptable acids or bases. 10. The compound of formula (I) according to claim 1, which is 1-methyl-3-propylamino-β-carboline-3-carboxamide. It and its addition salts with pharmaceutically acceptable acids or bases. 11. The compound of formula (I) according to claim 1, which is 3-(2-guanidino)-ethyl-β-carboline-3-carboxamide. It and its addition salts with pharmaceutically acceptable acids or bases. 12. The compound of formula (I) according to claim 1, which is 3-(3-guanidino)-propyl-β-carboline-3-carboxamide. It and its addition salts with pharmaceutically acceptable acids or bases. 13. The compound of formula (I) according to claim 1, which is 3-(6-guanidino)-hexyl-β-carboline-3-carboxamide. It and its addition salts with pharmaceutically acceptable acids or bases. 14. The compound of formula (I) according to claim 1, which is 3-(12-guanidino)-dodecyl-β-carboline-3-carboxamide. It and its addition salts with pharmaceutically acceptable acids or bases. 15. The compound of formula (I) according to claim 1, which is 1-methyl-3-(2-guanidino)-ethyl-β-carboline-3-carboxamide. It and its addition salts with pharmaceutically acceptable acids or bases. 16. The compound of formula (I) according to claim 1, which is 1-methyl-3-(3-guanidino)-propyl-β-carboline-3-carboxamide. It and its addition salts with pharmaceutically acceptable acids or bases. 17. The compound of formula (I) according to claim 1, which is 1-methyl-3-(6-guanidino)-hexyl-β-carboline-3-carboxamide. It and its addition salts with pharmaceutically acceptable acids or bases. 18. The preparation method of the compound of formula (I), characterized in that the compound of (II) is used as a raw material: wherein Ra, Rb, Rc, Rd and _R3 are as defined above. According to the conventional ester-forming conditions in organic synthesis, the compound of formula (III) can be obtained with methanol. According to the Pictet-Spengler reaction condition in organic synthesis, make this formula (III) compound react with (IV) compound: R ₂ CHO (IV) wherein R ₂ is as defined in formula (I). The compound of formula (V) is obtained: wherein Ra, Rb, Rc, Rd, _R2 and _R3 are as defined above. Be subjected to the effect of oxidizing agent commonly used in organic synthesis, obtain formula (VI) compound

wherein Ra, Rb, Rc, Rd, _R2 and _R3 are as defined above. According to conventional amidation conditions, process with formula (VII) compound: R _4a NHR _4b (VII) wherein R _4a and R _4b are as defined above. A specific example of a compound of formula (I/a) is obtained, i.e. a compound of formula (I):

wherein Ra, Rb, Rc, Rd, _R2 , _R3 , _R4a and _R4b are as defined above. The compound of formula is placed under the action of the compound of formula (VIII): [H ₂ NC(SCH ₃ )=NH] ₂ ·H ₂ SO ₄ (VIII) yields a compound of formula (I/b), a specific example of a compound of formula (I): wherein Ra, Rb, Rc, Rd, _R2 , _R3 and _R4b are as defined above. 19. Pharmaceutical composition comprising at least one compound of formula (I) according to any one of claims 1 to 17 as active ingredient, alone or in combination with one or several pharmaceutically acceptable, inert, nontoxic excipients form or carrier. 20. The pharmaceutical composition according to claim 19, comprising at least one active ingredient according to any one of claims 1 to 18, for anti-AIDS, anti-cancer treatment. 21. Pharmaceutical composition according to claim 19, comprising at least one active ingredient according to any one of claims 1 to 18, for use as an antioxidant in foods or pharmaceuticals and in antioxidant therapy in diseases.