CN1127335C - AIDS resisting medicine and its application - Google Patents

Abstract

The present invention relates to a medicine for resisting AIDS, which comprises a carrier for a formula (I) compound trans-1 nitro 2-thiophenyl-ethylene for resisting AIDS effective dose and medicine, and/or an excipient. The present invention simultaneously provides a reverse transcriptase inhibitor which comprises a formula (I) compound trans-1 nitro 2-thiophenyl-ethylene, a medicinal carrier and/or an excipient; the present invention provides an application of the formula (I) compound trans-1 nitro 2-thiophenyl-ethylene to preparation of medicine for resisting AIDS and an application of the trans-1 nitro 2-thiophenyl-ethylene to preparation of reverse transcriptase inhibitor medicine.

Description

Anti-AIDS drug and application thereof

The invention belongs to the pharmaceutical composition technical field, particularly, relate to a kind of anti-AIDS drug and the application in preparation reverse transcriptase inhibitors, anti-AIDS drug thereof.

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 as effective ingredient at the report aspect the anti-AIDS, the also report of the application of this chemical compound in preparation reverse transcriptase inhibitors medicine not.

The object of the present invention is to provide the medicine of anti-AIDS, anti--1 nitro 2-thiophenyl of formula (I) chemical compound-ethylene that wherein contains the anti-AIDS effective dose is used and pharmaceutically suitable carrier and/or excipient, provides this chemical compound in preparation reverse transcriptase inhibitors medicine and the application in the preparation anti-AIDS drug.

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

A kind of medicine of anti-AIDS, anti--1 nitro 2-thiophenyl of formula (I) chemical compound-ethylene that wherein contains the anti-AIDS effective dose is used and pharmaceutically suitable carrier and/or excipient.

A kind of reverse transcriptase inhibitors is provided simultaneously, has wherein contained anti--1 nitro 2-thiophenyl-ethylene of the disclosed formula of claim 1 (I) chemical compound and pharmaceutically suitable carrier and/or excipient.

The present invention also provides application and formula (I) chemical compound anti--1 nitro 2-thiophenyl-ethylene the application in preparation reverse transcriptase inhibitors medicine of anti--1 nitro 2-thiophenyl-ethylene of formula (I) chemical compound in the preparation anti-AIDS drug.

When of the present inventionization contains thing 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, the pharmacological action result of the pharmaceutical composition that will form with formula of the present invention (I) chemical compound racemization-2-methyl isophthalic acid-1,2,3,4-Tetrahydrooxonaphthalene-2-nitroethylene (hereinafter to be referred as KIBL-43) and with pharmaceutical carrier or excipient illustrates essence of the present invention below, but content of the present invention is not limited thereto.

1, anti--1 nitro 2-thiophenyl-ethylene is to the inhibitory action of HIV-1 reverse transcriptase

(1) experimental technique

(Reverse Transcriptase RT) measures on-radiation (non-radioactive) test kit available from Roche company to reverse transcriptase.With compound dissolution in DMSO.Take out in the test kit and react bar, every hole drips 20 μ l HIV-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.Compound concentrations 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 l ABTS substrate reactions liquid.Room temperature reaction 15-30 minute.The Bio-TekElx800 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: (seeing Figure of description 1)

Instead-1 nitro 2-thiophenyl-ethylene is to the inhibition activity data of HIV-1RT

μg/ml 0.288 1.44 7.2 36 180 EC ₅₀

Suppression ratio %-4.8-3.6 64.9 95.9 94.4 5.0 μ g/ml

2, anti--1 nitro 2-thiophenyl-ethylene is to the toxicity test of C8166 cell

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

(2) preparation of sample: is the concentration of 1000 μ g/mL with anti--1 nitro 2-thiophenyl-ethylene sample 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) that suppresses the 50%C8166 cell.(seeing Figure of description 2).

Instead-1 nitro 2-thiophenyl-ethylene is to toxicity test μ g/ml 15.6 31.3 62.5 125 250 500 CC50 suppression ratio %-17.3-10.3-6.5 30.7 78.3 104.2 155 μ g/ml of C8166 cell

3, anti--1 nitro 2-thiophenyl-ethylene to the therapeutic index of HIV (Selective Index, SI)

SI＝CC50÷EC50＝155μg/ml÷5.0μg/ml＝31

Can confirm from above-mentioned experimental result: anti--1 nitro 2-thiophenyl-ethylene of the present invention is inhibited to hiv reverse transcriptase, and it is low to the toxicity of C8166 cell, and the selection index height can be 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 Ono N et al, and J.Org.Chem., 1986,51:2139 synthesizes instead-1

Nitro 2-thiophenyl-ethylene:

(1) preparation of 1-nitro-2-thiophenyl-ethane is with 1-ethyoxyl-2-nitroethane (532mg, 4mmoL), (440nmg is dissolved in the acetonitrile 4mmoL) that (10mL adds triethylamine (410mg in 0 ℃ to thiophenol, 4mmoL), in stirring at room 30 minutes, in this solution impouring 1M hydrochloric acid, then with ethyl acetate extraction, after reclaiming solvent, get 1-nitro-2-thiophenyl-ethane 700mg (yield 95%).NMR(CDCl ₃)2.43(t，J＝8Hz，2H)，4.56(t，J＝8Hz，2H)，7.58(m，5H)。

(2) preparation of KIBL-43

Nitro-2-thiophenyl-ethane (630mg, 3.4mmoL) be dissolved in the 5mL dichloromethane, add thionyl chloride (0.290mL in 0 ℃, 3.6mmoL) under room temperature, stirred 5 minutes, remove volatile material and get residue, this residue is dissolved in the dichloromethane, under room temperature, add triethylamine (350mg, 3.6mmoL) stirred 30 minutes in 0 ℃, in this mixed liquor impouring water, with 1M salt pickling organic layer, boil off the organic layer solvent, get chemical compound 1-nitro-2-thiophenyl-ethane 500mg (yield 83%) IR 1505cm through the preparation Thin-layer separation ^-1NMR (CDCl ₃) 6.67 (d, J=12Hz, 1H), 7.24-7.56 (m, 5H), 8.19 (d, J=12Hz, 12Hz, 1H).

Structure: as shown at right

Molecular formula: C ₃H ₇NO ₂S

Character: yellow little needle

With synthetic anti--1 nitro 2-thiophenyl-ethylene and excipient weight ratio be 9: 1 ratio adding excipient, make powder.

Embodiment 2:

Method by embodiment 1 makes instead-1 nitro 2-thiophenyl-ethylene earlier, is that 5: 1 ratio adds excipient in itself and excipient weight ratio, makes powder.

Embodiment 3:

Method by embodiment 1 makes instead-1 nitro 2-thiophenyl-ethylene earlier, is that 5: 1 ratio adds excipient, pelletizing press sheet in itself and excipient weight ratio.

Embodiment 4:

Method by embodiment 1 makes instead-1 nitro 2-thiophenyl-ethylene earlier, and it is dissolved in the sterile water for injection, and stirring makes molten, filters with aseptic suction funnel, and aseptic again fine straining is sub-packed in the 2ml ampoule, and aseptic sealing by fusing gets injectable powder behind the frozen drying.

Embodiment 5:

Method by embodiment 1 makes instead-1 nitro 2-thiophenyl-ethylene earlier, and the oral liquid method for making is made oral liquid routinely.

Embodiment 6;

Method by embodiment 1 makes instead-1 nitro 2-thiophenyl-ethylene earlier, and the injection method for making adds the injection water routinely, fine straining, and injection is made in the embedding sterilization.

Embodiment 7:

Method by embodiment 1 makes instead-1 nitro 2-thiophenyl-ethylene earlier, is that 5: 1 ratio adds excipient in itself and excipient weight ratio, makes capsule.

Embodiment 8:

Method by embodiment 1 makes instead-1 nitro 2-thiophenyl-ethylene earlier, is that 3: 1 ratio adds excipient in itself and excipient weight ratio, makes capsule.

Claims (2)

1, a kind of medicine of anti-AIDS wherein contains anti--1 nitro 2-thiophenyl-ethylene of formula (I) chemical compound and the pharmaceutically suitable carrier and/or the excipient of anti-AIDS effective dose.

2, the application of anti--1 nitro 2-thiophenyl-ethylene of formula (I) chemical compound in the preparation anti-AIDS drug.