CN1122510C - Medicinal composition for treating AIDS and its application - Google Patents

Abstract

The present invention relates to a medical composition for treating AIDS, which comprises a formula (I) compound (dl-2-methyl-1-tetrahydronaphthalenone-2-nitroethylene) of effective dose for treating AIDS, a medicinal carrier and/or an excipient. The present invention also relates to an application of the medicine compound for preparing revertase inhibitor medicine and an application of the medicine compound for preparing medicine for resisting AIDS.

Description

A kind of medical composition and its use for the treatment of acquired immune deficiency syndrome (AIDS)

The invention belongs to technical field of pharmaceuticals, specifically, relate to a kind of pharmaceutical composition for the treatment of acquired immune deficiency syndrome (AIDS) and in preparation reverse transcriptase inhibitors medicine and the application in the preparation anti-AIDS drug.

Acquired immune deficiency syndrome (AIDS) (acquired immunodeficiency syndrome is called for short acquired immune deficiency syndrome (AIDS) for Acquired Immuno-deficiency Syndrome, AIDS) is that the world today mainly threatens one of the deadly disease that can not effect a radical cure of human health.The pathogen that causes acquired immune deficiency syndrome (AIDS) is that (Human Immunodeficiency Virus, HIV), nineteen eighty-three obtains to separate conclusive evidence human immune deficiency virus, two kinds of hypotype HIV-1 and HIV-2 is arranged, some variants.The HIV reproductive process roughly can be divided nine steps: adsorb, penetrate, shelling, early protein are synthetic, the duplicating of virus gene genome nucleic acid, late protein is synthetic, nucleocapsid assembling, virion maturation, release etc.In theory, each step in the said process can both be as the target spot of screening inverase.And each step has all been found many corresponding inhibitor, and dissimilar chemical compounds is studied in succession and is used for AIDS virus resisting.Yet wherein virus gene genome nucleic acid duplicate with proteinic synthetic be the step of HIV breeding most critical, and need the participation of some enzyme-specific, so the focus of the development of inverase mainly concentrates on the inhibitor aspect of seeking these enzyme-specifics, comprise reverse transcriptase (RT) inhibitor (it suppresses HIV from the mRNA transcription DNA), the protein synthesis enzyme inhibitor, reverse transcription starts the factor (TAT) inhibitor etc.The research of anti-HIV has been carried out to about 20000 chemical compounds in the whole world, has successfully developed 14 inverases of reverse transcriptase inhibitors and protease inhibitor at present.As reverse transcriptase inhibitors (AZT, DDC, DDI, D4T, 3TC, Nevirapine, Delavirdine, Efavinavir) and protease inhibitor medicine (Sequanavir, Ritonavir, Indinavir, Nelfinavir, Amprenavir) though be approved for clinically, objectively prolonged the life of HIV sufferers, simultaneously again with serious toxic and side effects such as bone marrow depression and appearance forgetful etc. and that cause disease-resistant strain.Thereby be badly in need of developing the novel drugs of newtype anti HIV-1 virus clinically with different chemical structures and mechanism of action.From Chinese herbal medicine, find natural HIV (human immunodeficiency virus)-resistant activity chemical compound or guide's thing---be importance and the very active field of studying both at home and abroad at present.So far find that more than 100 kind of native compound has the HIV activity, belong to sesquiterpene, diterpene, triterpene, steroidal, flavone, coumarin, peptide class, alkaloid, polysaccharide, trichosanthin etc., wherein active stronger as glycyrrhizin, hypericin, curcumin, soyasaponins, camptothecine, castanospermine, lentinan, trichosanthin etc.

So far, do not contain in the prior art formula (I) chemical compound racemization-2-methyl isophthalic acid-1,2,3,4-Tetrahydrooxonaphthalene-2-nitroethylene as effective ingredient at the report of treatment aspect the acquired immune deficiency syndrome (AIDS), the also not application of this chemical compound in preparation reverse transcriptase inhibitors medicine and the report of the application in the preparation anti-AIDS drug.

The object of the present invention is to provide the pharmaceutical composition that is used for the treatment of acquired immune deficiency syndrome (AIDS), formula (I) chemical compound racemization-2-methyl isophthalic acid-1,2,3,4-Tetrahydrooxonaphthalene-2-nitroethylene and pharmaceutical carrier and/or the excipient that wherein contains treatment acquired immune deficiency syndrome (AIDS) effective dose provides this pharmaceutical composition in preparation reverse transcriptase inhibitors medicine and the application in the preparation anti-AIDS drug.

In order to realize above-mentioned purpose of the present invention, the invention provides following technical scheme:

The pharmaceutical composition that is used for the treatment of acquired immune deficiency syndrome (AIDS) wherein contains formula (I) chemical compound racemization-2-methyl isophthalic acid-1,2,3,4-Tetrahydrooxonaphthalene--2-nitroethylene and pharmaceutically suitable carrier and/or the excipient for the treatment of the acquired immune deficiency syndrome (AIDS) effective dose.

The present invention also provides application and aforementioned pharmaceutical compositions the application in preparation anti-AIDS drug of aforementioned pharmaceutical compositions in preparation reverse transcriptase inhibitors medicine simultaneously.

When The compounds of this invention is used as medicine, can directly use, perhaps use with the form of pharmaceutical composition.This pharmaceutical composition contains 0.1-99%, is preferably the The compounds of this invention of 0.5-90%, and all the other are acceptable on the materia medica, to nontoxic and inert pharmaceutically suitable carrier of humans and animals and/or excipient.

Described pharmaceutical carrier or excipient are that one or more select solid, semisolid and liquid diluent, filler and pharmaceutical preparation adjuvant.Pharmaceutical composition of the present invention is used with the form of per weight dose.But two kinds of form administrations of medicine oral administration of the present invention and injection (quiet notes, intramuscular injection).

In order to understand essence of the present invention better, below will be with formula of the present invention (I) chemical compound racemization-2-methyl isophthalic acid-1,2,3,4-Tetrahydrooxonaphthalene--the pharmacological action result of 2-nitroethylene (hereinafter to be referred as KIBL-45) and the pharmaceutical composition formed with pharmaceutical carrier or excipient thereof illustrates essence of the present invention, but content of the present invention is not limited thereto.

1, racemization-2-methyl isophthalic acid-1,2,3,4-Tetrahydrooxonaphthalene--the 2-nitroethylene is to the inhibitory action of HIV-1 reverse transcriptase

(1) experimental technique

(Reverse Transcriptase RT) measures reverse transcriptase, and on-radiation (non-radioactive) test kit is available from Roche company.With compound dissolution in DMSO.Take out in the test kit and react bar, every hole drips 20 μ lHIV-1RT solution, 13 μ l lysis buffers, and 7 μ l are dissolved in the chemical compound of DMSO, and 20 μ l contain the reaction buffer of template and nucleotide.The whole Nong degree of chemical compound is 210 μ g/ml.Foscarnet positive control hole is set and does not contain the negative control hole of chemical compound.Put 37 ℃ 1 hour.After washing 5 times, every hole adds the anti-DIG-POD antibody of 200 μ l working solution.37 ℃ of incubations 1 hour wash 5 times.Every hole drips 200 μ lABTS substrate reactions liquid.Room temperature reaction 15-30 minute.Bio-Tek Elx800 microplate reader is measured OD value (405/490nm).Be judged to be the primary dcreening operation positive with the chemical compound that suppresses RT activity＞50%.The primary dcreening operation positive compound is carried out doubling dilution with lysis buffer, and each concentration is established two multiple holes.Measure it as stated above to the active inhibition of RT.Calculate the compound concentration (EC50,50%effective concentration) that suppresses HIV-1RT activity 50%.

(2) result's (seeing Figure of description 1)

Racemization-2-methyl isophthalic acid-1,2,3,4-Tetrahydrooxonaphthalene--2-nitroethylene is to the inhibition activity data of HIV-1RT

μg/ml 0.288 1.44 7.2 36 180 IC50

Suppression ratio %-6.3-3.4 58.9 99.0 103.3 6.3 μ g/ml

2, racemization-2-methyl isophthalic acid-1,2,3,4-Tetrahydrooxonaphthalene--the 2-nitroethylene is to the toxicity test of C8166 cell

(1) assay method: adopt the mensuration of mtt assay to sample CC50

(2) preparation of sample: racemization-2-methyl isophthalic acid-1,2,3,4-Tetrahydrooxonaphthalene--2-nitroethylene sample is the concentration of 1000 μ g/mL with the RPMI-1640 dilution, with 0.22 μ M membrane filtration.

(3) experimental implementation

Get the every hole of plate+100 μ L culture medium and carry out 2 times of gradient dilutions+100 μ L C8166 cells (3 * 10 ⁵/ mL), cultivate 72 hours (37 ℃ of CO ₂5%), sucking-off supernatant 100 μ L+20 μ L MTT continue 4 hours (37 ℃ of CO of incubation ₂5%) μ L+100,10%SDS is put in the incubator spend the night (18 hours).

(4) OD pH-value determination pH

Measure OD value wavelength 490nM with the Elx-800 microplate reader.

(5) experimental result; The inhibition % that calculates variable concentrations leads, calculates the toxicity value (CC50,50%cytoxic concentration) (seeing Figure of description 2) that suppresses the 50%C8166 cell.

Racemization-2-methyl isophthalic acid-1,2,3,4-Tetrahydrooxonaphthalene--2-nitroethylene is to the toxicity test μ g/ml 15.6 31.3 62.5 125 250 500 CC50 suppression ratio %-4.1 15.5 38.9 55.4 71.3 90.0 101 μ g/ml of C8166 cell

3, racemization-2-methyl isophthalic acid-1,2,3,4-Tetrahydrooxonaphthalene--2-nitroethylene to the therapeutic index of HIV (Selective Index,

SI)SI＝CC50÷EC50＝101μg/ml÷6.3μg/ml＝16

Can confirm from above-mentioned experimental result: racemization of the present invention-2-methyl isophthalic acid-1,2,3,4-Tetrahydrooxonaphthalene--the 2-nitroethylene is inhibited to hiv reverse transcriptase, it is low to the toxicity of C8166 cell, the selection index height, it is used for and reverse transcriptase diseases associated such as acquired immune deficiency syndrome (AIDS) etc.

Fig. 1 is the inhibition activity data figure of anti--1 nitro 2-thiophenyl-ethylene to HIV-1RT;

Fig. 2 is the toxicity test figure of anti--1 nitro 2-thiophenyl-ethylene to the C8166 cell.

Further flesh and blood of the present invention is described below in conjunction with embodiment, but content of the present invention is not limited thereto.

Embodiment 1:

According to document Node M.et al., Synthesis, 1987,8:729 carries out synthesising racemation-2-methyl isophthalic acid-1,2,3,4-Tetrahydrooxonaphthalene--2-nitroethylene:

2-methyl isophthalic acid-1,2,3,4-Tetrahydrooxonaphthalene 32mg (0.2mmoL) is dissolved in the 10mL oxolane, under nitrogen protection, adds diisopropylamine 34.46 μ L (0.2mmoL), drip n-BuLi 185 μ L (1.62mmoL/mL) at 0 ℃.In 0 ℃ of stirring 30mm, add Nitroenamine 47.4mg (being dissolved in the 5mL oxolane), in 1.5 hours, rise to room temperature, solution impouring NH from 0 ℃ gradually ₄In the Cl saturated aqueous solution, after the ethyl acetate base extraction, this reactant gets racemization-2-methyl isophthalic acid-1,2,3,4-Tetrahydrooxonaphthalene-2-nitroethylene 43mg (yield 93%) through the preparation Thin-layer separation.IR：1685，1505，1530，1460，1350.UV：207(24000)，250(16000). ¹H-NMR(CDCl ₃)：1.44(s，3H)；2.23(m，2H)；3.04(t，2H，J＝6.9)；6.94，7.53(Abq，2H，J＝14.0)；7.08-7.60(m，3H)；8.04(d，1H，J＝7.0)Node?M.et?al.，Synthesis，1987，8：729

Racemization-2-methyl isophthalic acid-1,2,3,4-Tetrahydrooxonaphthalene--2-nitroethylene structured data

Structure: as shown at right

Molecular formula: C ₁₃H ₁₃NO ₃

With synthetic racemization-2-methyl isophthalic acid-1,2,3,4-Tetrahydrooxonaphthalene--2-nitroethylene and excipient weight ratio are that 9: 1 ratio adds excipient, make powder.

Embodiment 2:

Method by embodiment 1 makes racemization-2-methyl isophthalic acid-1,2,3,4-Tetrahydrooxonaphthalene--2-nitroethylene earlier, is that 5: 1 ratio adds excipient in itself and excipient weight ratio, makes powder.

Embodiment 3:

Method by embodiment 1 makes racemization-2-methyl isophthalic acid-1,2,3,4-Tetrahydrooxonaphthalene--2-nitroethylene earlier, is that 5: 1 ratio adds excipient, pelletizing press sheet in itself and excipient weight ratio.

Embodiment 4:

Method by embodiment 1 makes racemization-2-methyl isophthalic acid-1,2,3,4-Tetrahydrooxonaphthalene--2-nitroethylene earlier, and it is dissolved in the sterile water for injection, and stirring makes molten, filter with aseptic suction funnel, aseptic again fine straining is sub-packed in 2 ampoules, and aseptic sealing by fusing gets injectable powder behind the frozen drying.

Embodiment 5:

Method by embodiment 1 makes racemization-2-methyl isophthalic acid-1,2,3,4-Tetrahydrooxonaphthalene--2-nitroethylene earlier, and the oral liquid method for making is made oral liquid routinely.

Embodiment 6:

Method by embodiment 1 makes racemization-2-methyl isophthalic acid-1,2,3,4-Tetrahydrooxonaphthalene--2-nitroethylene earlier, adds the injection water routinely, fine straining, and injection is made in the embedding sterilization.

Embodiment 7:

Method by embodiment 1 makes racemization-2-methyl isophthalic acid-1,2,3,4-Tetrahydrooxonaphthalene--2-nitroethylene earlier, is that 5: 1 ratio adds excipient in itself and excipient weight ratio, makes capsule.

Embodiment 8:

Method by embodiment 1 makes racemization-2-methyl isophthalic acid-1,2,3,4-Tetrahydrooxonaphthalene--2-nitroethylene earlier, is that 3: 1 ratio adds excipient in itself and excipient weight ratio, makes capsule.

Claims (3)

1, the pharmaceutical composition that is used for the treatment of acquired immune deficiency syndrome (AIDS) wherein contains formula (I) chemical compound racemization-2-methyl isophthalic acid-1,2,3,4-Tetrahydrooxonaphthalene--2-nitroethylene and pharmaceutically suitable carrier and/or the excipient for the treatment of the acquired immune deficiency syndrome (AIDS) effective dose.

2, the application of the described pharmaceutical composition of claim 1 in preparation reverse transcriptase inhibitors medicine.

3, the application of the described pharmaceutical composition of claim 1 in the preparation anti-AIDS drug.

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