CN101785777A

CN101785777A - Application of bufotalin std. in preparing medicament for treating lung tumors

Info

Publication number: CN101785777A
Application number: CN 201010136070
Authority: CN
Inventors: 侯惠民; 于垂亮
Original assignee: Shanghai Modern Pharmaceutical Engineering Research Center Co Ltd
Current assignee: Shanghai Modern Pharmaceutical Engineering Research Center Co Ltd
Priority date: 2010-03-30
Filing date: 2010-03-30
Publication date: 2010-07-28
Anticipated expiration: 2030-03-30
Also published as: CN101785777B

Abstract

The invention discloses an application of bufotalin std. in preparing a medicament for treating lung tumors. Proved by in vitro anti-tumor experiments of the bufotalin std., the concentration of medicaments of the animal lungs is higher than the concentration of effecting medicaments, and the bufotalin std. has stronger activity inhibition effect on human non-small cell lung cancer and other lung tumor cell stains, wherein the IC50 is 15-200ng/ml, and the bufotalin std. has good dose-response relationship. Shown as experimental results, the tumor growth inhibition effect of the bufotalin std. is superior to that of medicaments clinically applied at present, and the bufotalin std. has great development and application prospects.

Description

The application of bufotalien in preparation treatment lung tumors medicine

Technical field

The present invention relates to the new purposes of bufotalien, specifically relate to the application in preparation treatment lung tumors medicine of bufotalien and compositions thereof.

Background technology

Pulmonary carcinoma is modal lung primary malignant tumor, and most pulmonary carcinoma originate from the bronchial mucosa epithelium, so also claim lung bronchogenic carcinoma.Over nearly more than 50 years, countries in the world are industrially developed country particularly, and the sickness rate of pulmonary carcinoma and case fatality rate all rise rapidly, and the andropathy philtrum pulmonary carcinoma of dying from carninomatosis ranks first.

The transfer of each different organs late period can appear in pulmonary carcinoma, can cause corresponding symptom, usually brings great misery to patient, even threatens life.The most common clinical lung cancer metastasis is to brain, bone, liver and positions such as kidney and adrenal gland.

Pulmonary carcinoma has following two kinds of fundamental types: 1) patients with lung cancer of small cell lung cancer (SCLC) nearly 20% belongs to this type; 2) nonsmall-cell lung cancer (NSCLC) class, about 80% patients with lung cancer belongs to this type.

The treatment of pulmonary carcinoma at present has chemotherapy, radiotherapy and operative treatment etc., but because the distinctive biological behaviour of SCLC, and to the chemotherapy sensitivity, so the chemotherapy whole body therapeutic is very important.Many chemotherapeutic such as cyclophosphamide (CTX), ifosfamide (IFO), vinblastine (VLB), vincristine (VCR), chlormethine (HN2), etoposide (VP16) and oral capsule thereof, anthracycline drug, carboplatin (CBP) etc., the recent effective percentage of its single medicine is more than 40%.

Nonsmall-cell lung cancer (NSCLC), very inresponsive or insensitive to radiotherapy or chemotherapy, so once making a definite diagnosis, first-selected surgical resection therapy, postoperative according to circumstances give supplement therapy again.Resection rate is low on the one hand, even excision on the other hand, the probability of postoperative recurrence, transfer is also high, so for NSCLC, need Comprehensive Treatments such as operation, radiotherapy, chemotherapy, the quality of just might making the life better improves survival rate.May excise before all arts difficulty, postoperative have potential relapse and metastasis tendency, postoperative occur relapse and metastasis, stadium lost the excision chance evening, all needs chemotherapeutic treatment.

Because the progress of chemotherapy and increasing of effective chemotherapeutics, improved the recent effective percentage of NSCLC chemotherapy, but it is few to reach the case of CR.But in recent years, take a broad view of bibliographical information, changing the situation that can not prolong life cycle gradually transitivity NSCLC chemotherapy, but and some cases appears by chemotherapy proper extension life cycle, therefore chemotherapy has become indispensable means in the Comprehensive Treatment of NSCLC.The toxicity of certainly existing chemotherapy can not be ignored, as nephrotoxicity and gastrointestinal reaction etc.

Therapeutic effect, toxicity and the patient of existing chemotherapeutics all express very big expectation to the appearance of new chemotherapeutics to the toleration of chemotherapeutics.

Summary of the invention

Technical problem to be solved by this invention is to disclose the application of consistent bufotalien in preparation treatment lung tumors medicine, to satisfy the needs of clinical practice.

Bufotalien is a kind of chemical compound that extraction separation obtains from Chinese medicine Venenum Bufonis or Cutis Bufonis and Bufo siccus secretions, and its structural formula is:

Described bufotalien can adopt the method for bibliographical information be prepared (Yang Xiuwei etc., the people intestinal bacterium metabolism of cinobufacin and hydroxyl cinobufacin research [J]. Peking University's journal (medicine), 2001,33 (3): 199-204.; Zhang Ying etc. the chemical constituent of Bufo siccus skin [J]. Shenyang Pharmaceutical University's journal, 2007,24 (8): 484-487.), perhaps can adopt business-like product, as J﹠amp; K CHEMICAL LTD. company provides and is the product of 471-95-4 CAS number.

The inventor finds, described bufotalien has the function of excellent inhibition tumor growth, especially suppresses the function of lung tumors growth, can be used for the treatment of lung tumors.

Can be with the formation of compositions, by oral, intravenous injection or transdermal penetration mode, put on the patient who needs treatment, dosage is generally 0.05～3mg/ days/Kg, specifically can be determined by the doctor according to patient's the state of an illness, age etc.;

Described compositions comprises the bufotalien for the treatment of effective dose and pharmaceutically acceptable carrier, and described carrier comprises diluent, excipient such as water etc., filler is as starch, sucrose etc., binding agent, as cellulose derivative gelatin, polyvinylpyrrolidone etc., lubricant is as Pulvis Talci etc.

Contain in the compositions of bufotalien, the weight content of bufotalien is 0.01%～99.9%; Preferred weight content is 0.1%～99.0%.

Can adopt method well known in the art, will contain the compositions of bufotalien, preparation becomes injectable powder, granule, capsule, unguentum, cream, transdermal patch or other oral agents.

Through the bufotalien anticancer experiment in vitro, the applicant finds that the animal lung drug level all is higher than the onset drug level, and bufotalien has stronger inhibition activity, IC to people's nonsmall-cell lung cancer and other lung tumors cell strain ₅₀At 15～200ng/ml, and has dose-effect relationship preferably.Experimental result shows that the inhibition tumor growth effect of bufotalien is better than the medicine of clinical present application, has very great development application prospect.

The specific embodiment

Embodiment 1

The separation and purification of chemical compound bufotalien

Utilize pulverizer to break into fine particle after the Venenum Bufonis medical material broken into pieces, take by weighing 300g and place the 5L round-bottomed flask, add the 3L volumetric concentration and be 95% ethanol, heating in water bath backflow 6h, extracting liquid filtering; It is the same operation of 95% ethanol that medicinal residues add the 5L volumetric concentration again, extracts altogether 3 times, merges whole filtrates, when 55 ℃ of water-baths rotation is concentrated into 250ml, be transferred in the big pears type bottle and add the 500ml pure water again, add the 500ml chloroform, capping plug jolting and ultrasonic 2h place the layering of spending the night, and divide and get chloroform layer; The residue water layer adds the same operation of chloroform again, and coextraction repeatedly.Merge whole chloroform layers be evaporated to dried, solid.

Get chloroform extract, after the chloroform dissolving, add tlc silica gel G and mix sample, use the silica gel H upper prop, separate with the ethyl acetate gradient elution chromatography, obtain a plurality of streams parts with petroleum ether.The stream part that to contain bufotalien is that 45% methanol is that mobile phase is carried out the separated and collected respective components and is concentrated into driedly with volumetric concentration, can obtain the pure product of bufotalien.

Embodiment 2

Preparation bufotalien injection:

Prescription: (in part by weight)

Bufotalien 1-10 part,

5～100 parts of pharmaceutical grade ethanol

5～100 parts of Cremophor EL.

0.001～2 part of vitamin C.

Adopt known method, preparation becomes injection.

Embodiment 3

Tablet: (in part by weight)

Bufotalien 1-10 part

Starch 5-100 part

Hypromellose 0.1-10 part

Micropowder silica gel 0.01-0.5 part

Magnesium stearate 0.01-0.5 part

Adopt known method, preparation becomes tablet.

Embodiment 4

Bufotalien is to the inhibitory action of people's nonsmall-cell lung cancer A549

(Huang Yinjiu, wait " srb assay and the research of mtt assay screening anti-tumor medicine results relevance. " magazine biology (04): 13-16.) the identical srb assay of report, A549 carries out the vitro inhibition active testing to people's nonsmall-cell lung cancer to adopt document.

People's nonsmall-cell lung cancer A549 is provided by Chinese Academy of Sciences's Shanghai cell bank.

Trial drug: bufotalien is dissolved in the solution that is mixed with variable concentrations among methanol or the DMSO, and wherein, bufotalien is by J﹠amp; K CHEMICAL LTD provides.With the medicine of variable concentrations respectively application of sample survey its inhibition activity to the growth of people's nonsmall-cell lung cancer A549 cell, the application of sample amount is no more than 1%.Result of the test sees Table 1.

1, to the suppression ratio % of people's nonsmall-cell lung cancer A549 cell growth

Experimental result shows that bufotalien has stronger inhibitory action to people's nonsmall-cell lung cancer A549, has presented dose-effect relationship preferably, its IC ₅₀At 0.018 μ g/ml.

Embodiment 5

Bufotalien is determination of drug concentration in lung

Adopt the HPLC analytical method, adopt C ₁₈5 μ m, 250 * 4.6mm chromatographic column is with 0.5%KH ₂PO ₄-acetonitrile (with phosphoric acid adjust pH to 3.0) (60: 40) is as mobile phase, and flow velocity 1.0ml/min detects wavelength: UV296nm, column temperature: 40 ℃.

Bufotalien is mixed with the injection of 1mg/ml, gets mice, after accurately weighing, carry out the injection of mouse tail vein constant speed with the dosage of per 28 minutes input 4mg/kg.

After injection finished, mice was put to death in dislocation rapidly, takes out lung tissue, pure water blots after washing, and puts into the also accurate title of 10ml tool plug glass centrifuge tube after shredding with shears and decides weight, adds ethyl acetate 3.0ml, vortex vibration 2min, ultrasonic 20min, the centrifugal 10min of 3000R/min, (wherein blood plasma is put into-20 ℃ of refrigerators to go out the upper strata just, treat to go out the upper strata more just after freeze to solidify the bottom) dry up after, each adds 100 μ l dissolve with methanol, leaves standstill, and gets supernatant 20 μ l and carries out the HPLC analysis.

Analysis result sees Table 2.

Bufotalien distribution results (unit: μ g/g) in table 2 constant speed gasing injection mice (4mg/kg28min) lung tissue

By table 2 as seen, mouse lung inner tissue drug level can reach 0.872 ± 0.3165 μ g/g, is higher than the valid density of drug effect far away, has shown that bufotalien can well be distributed to lung tissue.

Bufotalien and the compositions that contains bufotalien can well be distributed to lung tissue, and can be issued on the active drug concentration in the stable state situation, and have very high growth inhibitory activity to the lung tumors tissue, therefore have the very big prospect that is developed to treatment lung tumors medicine.

Claims

1. the application of bufotalien in preparation treatment lung tumors medicine.

2. application according to claim 1 is characterized in that, described medicine comprises the bufotalien for the treatment of effective dose and pharmaceutically acceptable carrier.

3. application according to claim 2 is characterized in that, described medicine is injection, injectable powder, granule, capsule, unguentum, cream, transdermal patch or other oral agents.

4. according to claim 1,2 or 3 described application, it is characterized in that described lung tumors behaviour nonsmall-cell lung cancer.