CN101984962B

CN101984962B - Application of T-2 toxin to preparing drugs for treating brain tumors

Info

Publication number: CN101984962B
Application number: CN201010258664XA
Authority: CN
Inventors: 厉保秋
Original assignee: Shandong University
Current assignee: Shenzhen Carbon Cloud Intelligent Peptide Pharmaceutical Technology Co ltd
Priority date: 2009-03-03
Filing date: 2009-03-03
Publication date: 2011-12-07
Anticipated expiration: 2029-03-03
Also published as: CN101984962A

Abstract

The invention discloses new application of T-2 toxin, namely application of T-2 toxin to preparing drugs for treating brain tumors. The effective treatment amount of T-2 toxin is 0.1-20mg/Kg, preferably 0.5-8mg/Kg. The drugs are administrated by implantation, oral administration, intravenous injection, hypodermic injection and intramuscular injection, preferably intra-tumor injection. The inventor builds various solid tumor models and then examines the inhibiting effect of T-2 toxin on the tumors or tumor cells by carrying out intra-tumor injection or culture with cells on T-2 toxin. Experiments show that intra-tumor injection has confirmed curative effect on the tumors, especially the solid tumors hard to excised and drug administration through intra-tumor injection can further reduce the side effect range and degree of T-2 toxin, reduce the side effects of T-2 toxin and improve the inhibiting and killing effects of T-2 toxin on the tumor tissues.

Description

The application of T-2 toxin in the medicine of the preparation treatment cerebral tumor

Technical field

The present invention relates to the application of T-2 toxin in the medicine of the preparation treatment cerebral tumor.

Background technology

Tumor is one of common disease that threatens human health, be only second to cardiovascular disease, research about tumor has obtained bigger progress, developed the method such as various radiotherapies, chemotherapy of multiple treatment tumor, but all there are limitation in various medicines and therapy, be difficult to reach the effect of radical cure, and be prone to drug resistance and side effect.The tumor mortality rate still occupy first of the various diseases.The tool data show, China had 3,000,000 people to die from cancer in 2006, and tumor incidence still is in ascendant trend, and rejuvenation trend is arranged.In less than the time in 20 years, China's tumor incidence has risen 69% according to statistics, and mortality rate has risen 29.4%.Therefore, the new developing effective tumor treating medicaments of research is still the main direction of present oncotherapy.Solid tumor is generally adopted the method for excision, but excision is bigger to patient's damage, and is difficult to guarantee that excision is clean, after tumor tissues occurred soaking into adhesion, obscure boundary excision difficulty was higher, and some cerebral tissue solid tumor excision risk is very high.Radiotherapy also is one of common method in oncotherapy, but, it is comparatively serious to injury of human, often cause patient's system to descend, prognosis mala, be difficult to bear the side effect of chemotherapy for the part patient, and radiotherapeutic effect is also unsatisfactory, and cost issues also is one of reason of its application of restriction.Therefore the Drug therapy to tumor is still the method for the most generally using.

T-2 toxin, molecular weight are 466.122, and molecular formula is C ₂₄H ₃₄O ₉, structural formula is 4 β, 15-diacetoxy-8 α-(3-methoxyl group acyloxy)-3 Alpha-hydroxies-12, and the single-ended spore of 13-epoxy is mould-9-alkene, oxygen ring wherein and two keys are its active sites, the oxygen ring is opened or the reduction of two key all can make its toxicity descend.The T-2 toxin is a white, needle-shaped crystals, and fusing point is 151 ℃-152 ℃, is insoluble in water, is soluble in polar organic solvent, and it is quite stable at ambient temperature, places 6-7 or be heated to 100-120 ℃ of 1 hour toxicity not subtract.The T-2 toxin has ester group, becomes corresponding alcohol with the alkali treatment posthydrolysis.

The T-2 toxin is the strongest a kind of of the mould alkene of the single-ended born of the same parents family toxin poisoning that produces of Fusarium spp., mainly act on the vigorous histoorgan of cell division, as thymus, bone marrow, liver, spleen, lymph node, gonad and gastrointestinal mucosa etc., it is synthetic to suppress these organ cell's protein and DNA.In addition, find that also this toxin can cause dna single chain break in the lymphocyte.The T-2 toxin also can act on a plurality of positions of oxidative phosphorylation and cause that mitochondrial respiratory suppresses.And tumor tissues promptly has the splitted characteristics of cell transition, therefore, there is toxic action widely in tumor tissues.

Research to the T-2 toxin at present mainly concentrates on its toxicity research and Study on Pathogenicity to animal, and the research that the T-2 toxin is used for the treatment of solid tumors medicine still finds no report.

Summary of the invention

At above-mentioned prior art, the invention provides a kind of new purposes of T-2 toxin.

The present invention comes down to the application of T-2 toxin in preparation treatment solid tumor drugs.

Described solid tumor comprises Vipoma, lung tumor, hepatoma, renal cell carcinoma, the cerebral tumor, prostate tumor.

Effective therapeutic dose of described T-2 toxin is 0.1～20mg/Kg body weight, preferred 0.5～8mg/Kg body weight.

Described application is that the T-2 toxin is made the intratumor injection preparation

During administration, can be drug delivery implant, oral administration, intravenous administration, subcutaneous injection administration, administered intramuscular, preferred intratumor injection.

The present inventor has set up various solid tumors, mainly be the animal model or the cell model of cancer of pancreas, pulmonary carcinoma, hepatocarcinoma, renal cell carcinoma, the cerebral tumor, carcinoma of prostate, then the T-2 toxin investigated the inhibitory action of T-2 toxin to tumor or tumor cell by intratumor injection or with the mode of co-culture of cells.Experiment shows, the solid tumor that tumor especially is difficult to excise by intratumor injection has definite curative effect, and by the administration of intratumor injection mode, can further reduce the side effect scope and the degree of T-2 toxin, reduce its side effect and improve inhibition and lethal effect tumor tissues.

The specific embodiment

The present invention is further illustrated below in conjunction with test:

Embodiment 1:T-2 toxin is to the treatment of pancreatic cancer experimental study of effect

The human pancreas cancer fragment of tissue subcutaneous transplantation of diameter 2-3mm is arrived in the mice body, treat tumor length behind 6-7mm, will be dissolved in the dosage intratumor injection of the T-2 toxin of ethanol mixed liquor of normal saline according to the 2.5mg/kg body weight, matched group injecting normal saline.The the 1st, 5,9,14,19, the 24 day size with slide gauge or standard form measurement tumor measured lasting 24 days in the injection back.After treatment in 24 days, the gross tumor volume of T-2 toxin treated animal only is 52.19% of a matched group.

Embodiment 2:T-2 toxin is to the lung cancer therapy experimental study of effect

The Mice Bearing Lewis Lung Cancer cell is placed the complete culture solution that contains 10% hyclone and DMEM, at 37 ℃, 5%CO ₂Support in the case and cultivate, 2-3d changes a culture fluid.Cell is the monolayer adherence growth, and 0.5% trypsinization goes down to posterity.After total cellular score reached requirement, the trypan blue exclusion method was measured viable count greater than 95%, and regulating cell concentration is 5 * 10 ⁶/ ml, the standby C57BL/6 mouse inbred lines of cell suspension, male, 20,6～8 ages in week, body weight (18 ± 1) g.The armpit subcutaneous vaccination.Every inoculation 0.2ml, cell counting 5 * 10 ⁶Individual cell/ml.Mice is divided into 2 groups respectively at random, 10 every group.Matched group intratumor injection 2ml normal saline, experimental group will be dissolved in the dosage intratumor injection of the T-2 toxin of ethanol mixed liquor of normal saline according to the 2.0mg/kg body weight.Result of the test sees Table 1, and result of the test shows that the T-2 toxin has the obvious suppression effect to the growth of lung tumor.

Table 1 T-2 toxin is to the inhibitory action of pulmonary carcinoma

**P＜0.01

Embodiment 3:T-2 toxin is to the liver cancer treatment experimental study of effect

Adopt mice H ₂₂Hepatoma cell strain, the ICR mouse peritoneal goes down to posterity.Aseptic condition extracts the abdominal cavity guarantor down and planted eugonic H 7-9 days ₂₂Hepatoma carcinoma cell, regulating cell concentration with normal saline is 5 * 10 ⁶/ ml, 20 of cleaning agent ICR mices, in age in 6-8 week, male, 18-22g is divided into 2 groups at random, 10 every group.Equal left axil subcutaneous vaccination tumor cell suspension 0.2ml.Inoculate after 7 days, matched group intratumor injection 2ml normal saline, experimental group will be dissolved in the dosage intratumor injection of the T-2 toxin of ethanol mixed liquor of normal saline according to the 2.0mg/kg body weight.With vernier caliper measurement tumor major diameter (L) and minor axis (W), calculate gross tumor volume and inhibition rate of tumor growth next day of after the grouping administration, the results are shown in Table 2.Result of the test shows that the T-2 toxin has the obvious suppression effect to the growth of liver tumor.

Table 2 T-2 toxin is to the inhibitory action of pulmonary carcinoma

**P＜0.01

Embodiment 4:T-2 toxin is to the renal cell carcinoma treatment experimental study of effect

Adopt pregnant clear cell cancerous cell line RCC-949, adjusting cell concentration behind the In vitro culture is 1 * 10 ⁷/ ml, it is subcutaneous that 0.2ml injects the right lower limb dorsal part of nude mice, inoculates after 7 days, takes out tumor tissue and be cut into 1mm ³Fritter carries out going down to posterity between Mus, and getting the 26th generation cell adjustment cell concentration is 1 * 10 ⁷/ ml, 1% pentobarbital sodium intraperitoneal injection of anesthesia (50 μ g/10g) nude mice, left side lower back longitudinal incision, cut off Musclar layer, expose left kidney, carry out injection under the kidney peplos, 0.1ml/ only, inoculate after 7 days, matched group intratumor injection 2ml normal saline, experimental group will be dissolved in the dosage intratumor injection of the T-2 toxin of ethanol mixed liquor of normal saline according to the 2.0mg/kg body weight.With vernier caliper measurement tumor major diameter (L) and minor axis (W), calculate gross tumor volume and inhibition rate of tumor growth next day of after the grouping administration.Result of the test shows that the T-2 toxin reaches 48.35% to the growth inhibition ratio of renal cell carcinoma.

Embodiment 5:T-2 toxin is to the experimentation of cerebral tumor therapeutical effect

The trophophase human brain tumour SF763 cell of taking the logarithm, it is 1 * 10 that normal saline is regulated concentration ⁶/ ml inoculates the BALB/cA mice, and the inoculation volume is 0.025ml, and the inoculation point is the fixed point of an equilateral triangle, and the base is the line of auris dextra and right eye, and height is about 5mm, and the inoculation degree of depth is about 2mm.With mice with 4.35% trichlorine chloral hydrate anesthesia after, alcohol disinfecting is inoculated with syringe.Inoculate after 7 days, mice is divided into 2 groups respectively at random, 10 every group.Matched group intratumor injection 2ml normal saline, experimental group will be dissolved in the dosage intratumor injection of the T-2 toxin of ethanol mixed liquor of normal saline according to the 2.0mg/kg body weight.Observe and the record death time of animal, compute The average survival time natural law and increase in life span, increase in life span the results are shown in Table 3 with the percent calculating divided by matched group The average survival time natural law gained of the difference of experimental group The average survival time natural law and matched group The average survival time natural law.Result of the test shows that the T-2 toxin has the obvious suppression effect to the growth of the cerebral tumor.

Table 3 T-2 toxin is to the therapeutical effect of the cerebral tumor

**P＜0.01

Embodiment 6:T-2 toxin is to the prostate cancer therapy experimental study of effect

Carcinoma of prostate PC-3 cell culture adds 100uml in the RPM11640 culture medium that contains 10% hyclone ^-1Penicillin and 100mgL ^-1Streptomycin, the conventional cultivation got exponential phase of growth about 5 * 10 under 37 ℃ of conditions that contain 5% carbon dioxide ⁶/ ml passage and being inoculated in 24 orifice plates adds the T-2 toxin that the people is dissolved in the ethanol mixed liquor of normal saline after cultivating 12h.Significant growth inhibited effect appears in drug effect after 48 hours.Suppression ratio is 51.79%.

The preparation of embodiment 7:T-2 toxin intratumor injection preparation

Get T-2 toxin 60mg, be dissolved in the NaOH solution of 17ml1mol/L, add the 50ml propylene glycol, add water for injection at last to 100ml.

Claims

1.T-2 the application of toxin in the medicine of the preparation treatment cerebral tumor, it is characterized in that: effective therapeutic dose of described T-2 toxin is the 2.0mg/Kg body weight.

2. application according to claim 1 is characterized in that: be that the T-2 toxin is made the intratumor injection preparation.