Technical field

The present invention relates to a kind of antineoplastic combined medicament, specially refer to the antineoplastic combined medicament that is used to prepare the medicine for the treatment of glioma brain tumour, hepatocarcinoma and cervical cancer, belong to the cancer therapy drug technical field with enhancing and poison-reducing character.

Background technology

Glioma also claims glioma.To gliomatous treatment based on operative treatment, but because tumor is infiltrative growth, and no obvious border between cerebral tissue, except that infantile tumour the little and person that is positioned in the suitable position, be difficult to accomplish complete resection, generally all advocate Comprehensive Treatment, promptly postoperative cooperates with radiotherapy, chemotherapy etc., can delay recurrence and prolong life cycle.Glioma is one of main type of brain tumor, because the factor of blood brain barrier, main chemotherapeutics has chemotherapeutics such as temozolomide and teniposide, and these chemotherapeutics all have fat-solubility and can be by the character of blood brain barrier.But temozolomide (English Temozolomide) has the obvious toxic-side effects that comprises carcinogenic, teratogenesis and genotoxicity; Teniposide (English Teniposide) or similarly etoposide (English Etoposide) in the treatment cerebral tumor, also exist feel sick, digestive tract side effects such as vomiting, and bone marrow depression toxicity, unsuitable long-term and heavy dose of the use.Hepatocarcinoma also claims hepatocarcinoma, is a kind of invasive malignant tumor that has, and the back of healing is relatively poor, and it is popular B-mode closely related with infection with hepatitis C virus with China that development takes place for it.Because hepatocyte had been subjected to grievous injury when hepatocarcinoma took place, and caused treatment means very limited.Still do not have so far and formally get permission effective medicine.Treatment such as existing cancer treatment drug amycin, fluorouracil effective percentage is low excessively, and does not have obvious life prolongation effect.Cervical cancer accounts for 2% of malignant tumor, belongs to the higher gynecologic malignant tumor of sickness rate.At present, because the limitation of single therapy means, the treatment of cervical cancer is to be main means with operation, radiotherapy, chemotherapy.Cisplatin class (as carboplatin) commonly used, fluorouracil (as capecitabine) have certain curative effect, but have apparent in view toxic and side effects as chemotherapeutics.The research and development newtype drug, efficacy enhancing and toxicity reducing is the long-term and arduous work of tumor chemotherapeutic drug.

Teniposide is a kind of cell toxicity medicament of period specific, its mechanism of action is a DNA topoisomerase II specific inhibitor, clinical central nervous system's malignant tumor neuroblastoma that is used for, glioma and astrocytoma and metastatic tumor, and malignant lymphoma, the treatment of acute lymphoblastic leukemia.Teniposide is a micromolecular compound, and molecular weight is 656.6, and fat-soluble extremely strong, the logarithm value log of profit partition coefficient P ₁₀P is 1.96, is its key property that is easy to see through blood brain barrier treatment brain cell tumor.

The report of artemisine compounds to the cytotoxic activity of Ehrlich ascites cells just arranged in 1993: mtt assay records the half toxic concentration IC of arteannuin ₅₀Be 29.8 μ M, the IC of chemical compounds such as dihydroarteannuin, artesunate, Artemether and arteether ₅₀Then be 12.2-19.9 μ M[Woerdenbag H.J.et al.Cytotoxicity of artemisinin-related endoperoxidesto Ehrlich ascites tumor cells.J.Nat.Prod.1993; 56:849-56].Other has bibliographical information, and dihydroarteannuin (having another name called dihydroartemisinine) has very strong inhibitory action to human breast carcinoma MCF-7 cell, utilizes srb assay to record IC ₅₀Being 0.26 μ M, suppressing one of mechanism of MCF-7 cell proliferation, is that cell is trapped in the G0+G1 phase; Arteannuin only has faint inhibitory action to the MCF-7 cell proliferation, but artesunate has significant inhibitory effect.Utilize srb assay to record the half-inhibition concentration IC of artesunate to MCF-7 cell proliferation ₅₀Be 0.31 μ M, the apoptosis that causes at 1 μ M and directly cytotoxicity obviously be better than the effect [Lin Fang of 10 μ M arteannuin, Deng. (1) dihydroartemisinine is to human breast carcinoma MCF-7 cells in vitro inhibitory action. Chinese Journal of New Drugs 2002,11:934-936; (2) arteannuin and artesunate are to the comparative study of human breast carcinoma MCF-7 cells in vitro inhibitory action. Chinese herbal medicine 2003,34:347-349].Research is also found, through artesunate or dihydroarteannuin interaction in vitro 48 hours, four kinds of growth of tumour cell such as s, uterus choriocarcinoma jar cell, embryo muscular tumor RD cell, ovarian cancer HO-8910 cell are subjected to remarkable inhibition, the half-inhibition concentration IC that records with mtt assay ₅₀Be respectively 15.4-49.7 μ M (artesunate) or 8.5-32.9 μ M (dihydroarteannuin) (Chen H.H., et al.Inhibition of human cancer cellline growth and human umbilical vein endothelial cell angiogenesis by artemisinin derivatives invitro.Pharmacol Res 2003; 48:231-236).The derivant that arteannuin is described is potential antitumor drug.

The derivant of arteannuin, arteannuin, dihydroarteannuin, Artemether, arteether and artesunate particularly have been developed to into medicine, no matter be oral formulations, or injection and suppository, all proved the current specific drug that is used for the treatment of malaria by pharmacology and clinical research.But this class medicine also has the characteristic of two aspects---fat-soluble and safety fully is not applied to the research and development of antineoplastic combined medicament as yet.The characteristics of first aspect are, artemisinin derivative belongs to micromolecular compound and has strong especially fat-soluble.For example, the molecular weight of arteannuin, dihydroarteannuin and Artemether is 282.3,284.3,298.4 successively, the logarithm value log of profit partition coefficient P ₁₀P is 1.68,1.78 and 2.24 successively, guarantees to make it can pass through blood brain barrier smoothly, enters the central nervous system brain cell of unifying by blood circulation, helps it to suppress the growth of brain tumor cell, becomes one of a limited number of anti-cerebral tumor medicines.The characteristics of second aspect are, no matter the use of the derivant medicine of the arteannuin that the present invention relates at cellular level, still comprises the clinical patients treatment level in whole animal; No matter be according to the administration of malaria treatment standard or according to the treatment of cancer code requirement, under employed treatment consumption, all be proved to be good drug safety: toxic and side effects is all much smaller more than toxicity and toxic and side effects that conventional anti-tumor chemotherapeutic medicine brings.Therefore, derivant at the proof arteannuin has antitumor action, and can produce under drug effect addition or the synergistic situation of drug effect during with certain conventional chemotherapy of tumors medicine coupling, illustrate that with regard to having reason the two antineoplaston that share this routine chemotherapy of tumors medicine has potentiation or Attenuation, perhaps have the more satisfactory effect of not only potentiation but also attenuation.The research document of publishing has illustrated that the derivant of arteannuin is potential antitumor drug.The derivant of the said arteannuin of the present invention, promptly arteannuin, dihydroarteannuin, Artemether, arteether and artesunate be because all have sesquiterpene parent nucleus identical with arteannuin and peroxide bridge basic structure, and antitumor action is in various degree all arranged.

Summary of the invention

The object of the present invention is to provide a kind of antineoplastic combined medicament with enhancing and poison-reducing character.This composition of medicine is made up of a kind of derivant of teniposide and arteannuin, not only have the antineoplastic of enhancing effect, and it can also reduce the murder by poisoning of medicine to human body.

The technical solution adopted in the present invention is:

A kind of antineoplastic combined medicament with enhancing and poison-reducing character, it is made up of as active component a kind of derivant of teniposide and arteannuin, a kind of derivant that contains teniposide and arteannuin in pharmaceutical preparation simultaneously is as anti-tumor active ingredient, wherein the quality of teniposide accounts for the 1%-99% of mass fraction, preferred 10%-50%; The quality of a kind of derivant of arteannuin accounts for the 1%-99% of mass fraction, preferred 50%-90%.A kind of derivant of arteannuin is a kind of in arteannuin, dihydroarteannuin, Artemether, arteether and the artesunate; The preferred dihydroarteannuin that uses.A kind of derivant of active constituents of medicine teniposide and arteannuin is made pharmaceutical preparation.

In the pharmaceutical preparation of antitumor combination medicine, a kind of derivant that contains teniposide and arteannuin at least simultaneously that is to say as anti-tumor active ingredient, can also contain other anti-tumor active ingredients.

The present invention also discloses this anti-tumor compositions, and to be used to prepare malignant tumor be cancer, particularly the purposes of glioma brain tumour, hepatocarcinoma and cervical cancer medicine.

As active component, can make pharmaceutically acceptable dosage form with antineoplastic combined medicament disclosed by the invention, comprise injection, tablet, capsule, granule, slow releasing preparation, controlled release preparation and nanometer formulation, be applied to clinical.

The derivant that the present inventor has confirmed arteannuin has remarkable inhibitory action to the growth of kinds of tumor cells such as cervical cancer, glioma brain tumour, hepatocarcinoma, pulmonary carcinoma and breast carcinoma, and this is an important prerequisite condition of the present invention.

The derivant and the antineoplastic agent teniposide of arteannuin are united use, can on some tumor cells, produce the effect of inhibitory action " addition " and " working in coordination with ".

Different types of tumors, the generation development mechanism and the influence factor of its tumor cell there are differences, and the sensitivity of medicine also be there are differences.The invention discloses the antineoplastic combined medicament of forming by the derivant such as the dihydroarteannuin of teniposide and arteannuin, preferred development becomes the particularly pharmaceutical preparation of glioma and liver cell tumor of treatment cervical cancer, the cerebral tumor, is suitable for showing in treatment the efficacy enhancing and toxicity reducing effect most.Artemether, arteether and artesunate all pass through metabolism in the humans and animals body and biotransformation forms dihydroarteannuin, therefore, the said antineoplastic combined medicament of the present invention is meant the medicine that any one derivant in teniposide and arteannuin, dihydroarteannuin, Artemether, arteether and the artesunate is formed.

The present invention has following advantage with respect to prior art: the antineoplastic combined medicament of being made up of the derivant of teniposide and arteannuin has the efficacy enhancing and toxicity reducing effect when suppressing growth of tumour cell; Can be used to make malignant tumor is cancer, particularly the medicine of glioma brain tumour, hepatocarcinoma and cervical cancer.

The specific embodiment

Below in conjunction with embodiment the present invention is done further to describe in detail, it should be understood that these embodiment only are used for illustrating the present invention, do not limit the scope of the invention.

Embodiment 1 tumor cell kind and cell culture processes thereof

Cell strain and cell culture: test used cell and comprise s, glioma brain tumour SWO-38 cell, hepatocarcinoma HepG2 cell, pulmonary carcinoma LAC cell and breast carcinoma MCF-7 cell.The condition of culture of all cells is as follows: (37 ℃, saturated humidity, CO in carbon dioxide cell incubator ₂Content 5%), grow in and contain in 10% hyclone and the antibiotic RPMI-1640 culture fluid of 100U/mL.Per 3 to 4 days passages once.

Embodiment 2 dihydroarteannuins and teniposide are to the inhibitory action of growth of tumour cell

Drug effect: collect the cell that is in exponential phase, in culture fluid, be dispersed into every milliliter and contain 5 * 10 ⁴The individual cells suspension of cell.Cell suspension is inoculated in 96 porocyte culture plates by every hole 100 microlitres, cultivates 24 hours in carbon dioxide cell incubator.Discard original culture fluid, add culture fluid 100 microlitres that contain medicine to be tested, continue to cultivate 72 hours, observe the effect of medicine cell growth by per 3 Kong Weiyi detectable concentrations.Establish culture fluid blank and the not cell contrast of dosing processing in the experiment in addition.Measure the half-inhibition concentration of medicine cell growth with mtt assay.

Used mtt assay basic operation: after 68 hours, it is MTT solution 10 microlitres of 5 mg/ml that every hole adds concentration to the cell in 96 orifice plates through drug treating, and reaction is 4 hours in carbon dioxide cell incubator.Take out 96 porocyte culture plates, careful sucking-off culture fluid discards, in the hole residual pigementation with 150 microlitre dmso solutions after, measure optical density value in 490 nanometers with microplate reader.This moment, the optical density value in every hole was directly proportional with living cells quantity.

Cell survival rate (%)=(the blank group of experimental port optical density value-cultivation optical density value)/(the blank group of cell matched group optical density value-culture fluid optical density value) * 100%

The half-inhibition concentration IC of medicine cell growth ₅₀Be that cell survival rate is 50% o'clock a drug level.

The result is as shown in Table 1 and Table 2:

Table 1 dihydroarteannuin is to the inhibitory action of growth of tumour cell ^*

 

^*Cell growth inhibition test is triplicate at least, and the result is with meansigma methods (x) ± standard deviation (SD) expression; With solvent and culture fluid as the medicine blank.IC ₅₀: half-inhibition concentration (μ M)

Table 2 teniposide is to the inhibitory action of growth of tumour cell ^*

^*Cell growth inhibition test is triplicate at least, and the result is with meansigma methods (x) ± standard deviation (SD) expression; With solvent and culture fluid as the medicine blank.IC ₅₀: half-inhibition concentration (μ M)

Synergism or potentiation that embodiment 3 dihydroarteannuin coupling teniposides have when suppressing some growth of tumour cell

Experimentation is the same.The results are shown in Table 3:

Table 3 dihydroarteannuin (A) suppresses working in coordination with or potentiation of growth of tumour cell with teniposide (B) ^*

^*Cell growth inhibition test is triplicate at least, and the result is with meansigma methods (x) ± standard deviation (SD) expression; With solvent and culture fluid as the medicine blank.IC ₅₀: half-inhibition concentration (μ M)

Experimental result shows: the first, and with accounting for IC ₅₀9.4% dihydroarteannuin of quantity adds to account for IC ₅₀30% teniposide of quantity can produce half and suppress effect (curative effect addition or synergism) to the growth of s; With accounting for IC ₅₀3% dihydroarteannuin of quantity adds to account for IC ₅₀35% teniposide of quantity can produce half and suppress effect (curative effect synergism) to the growth of glioma brain tumour SWO-38 cell; With accounting for IC ₅₀15.7% dihydroarteannuin of quantity adds to account for IC ₅₀4.7% teniposide of quantity can produce half and suppress effect (curative effect synergism) to the growth of hepatocarcinoma HepG2 cell.The second, in the chemotherapy practice, when using artemisinin derivative, the dose that suitably reduces teniposide also can obtain needed antitumous effect, has so also just reduced the toxic and side effects that is caused by teniposide.

 

1. an antineoplastic combined medicament is characterized in that: the antineoplastic combined medicament that contains a kind of derivant of teniposide and arteannuin.

2. antineoplastic combined medicament according to claim 1 is characterized in that: a kind of derivant that contains teniposide and arteannuin is as anti-tumor active ingredient.

3. antineoplastic combined medicament according to claim 1 and 2 is characterized in that: a kind of derivant of described active component arteannuin is meant a kind of in arteannuin, dihydroarteannuin, Artemether, arteether and the artesunate preferably uses dihydroarteannuin.

4. according to any described antineoplastic combined medicament of claim 1-3, it is characterized in that: in described active component, the quality of teniposide accounts for the 1%-99% of mass fraction, preferred 10%-50%; The quality of a kind of derivant of arteannuin accounts for the 1%-99% of mass fraction, preferred 50%-90%.

5. according to any described antineoplastic combined medicament of claim 1-4, it is characterized in that: a kind of derivant of active constituents of medicine teniposide and arteannuin is made pharmaceutical preparation.

6. antineoplastic combined medicament according to claim 5 is characterized in that: in pharmaceutical preparation, a kind of derivant that contains teniposide and arteannuin at least simultaneously is as anti-tumor active ingredient.

7. according to any described antineoplastic combined medicament of claim 1-6, it is characterized in that: being used to prepare malignant tumor is cancer, particularly glioma brain tumour, hepatocarcinoma and cervical cancer medicine.

8. according to any described antineoplastic combined medicament of claim 1-6, it is characterized in that: described medicine is made into pharmaceutically acceptable dosage form, comprises injection, tablet, capsule, granule, slow releasing preparation, controlled release preparation and nanometer formulation.