CN105250303B - Fluoxastrobin is to the intervention effect of carcinoma of mouth and precancerous lesion - Google Patents

Abstract

Preparation of the present invention on a kind of new application of compound Fluoxastrobin (E) [2 [6 (2 cyano-benzene oxygen) pyrimidine, 4 base oxygen] phenyl] 3 methoxy-methyl acrylates (methyl (E) 2 { 2 [6 (2 cyanophenoxy) pyrimidin, 4 yloxy] phenyl } 3 methoxyacrylate) and for significantly inhibiting cancer cell of oral cavity belongs to antitumor drug research and development field.Fluoxastrobin acquisition methods are easy, chemical constitution is clear and definite, it is detected using the proliferation activity of 8 method oral cancer cell SCC15 and CAL27 of CCK and oral precancerous lesion DOK cells, experimental result shows apparent inhibited proliferation, the present invention provides experimental basis for the research and development of clinical treatment carcinoma of mouth new drug, can be applied to inhibit in the preparation that cancer cell of oral cavity is grown.

Description

Fluoxastrobin is to the intervention effect of carcinoma of mouth and precancerous lesion

Technical field

The present invention relates to a kind of new opplications of compound, particularly relate to a kind of new intervening oral cancer cell life Long compound.

Background technology

Carcinoma of mouth is the common malignant tumour of incidence, accounts for the 3% of whole body malignant tumour, occupies the 6th.Carcinoma of mouth Incidence it is worldwide in rising trend, age of onset rejuvenation and women increasing trend estimate the annual new hair in the whole world Case 27.4 ten thousand.Oral cavity is the important anatomical site of human body, carcinoma of mouth patient using appearance damage, eating difficulties and pain as There is a series of clinical manifestations such as difficulty in opening mouth, saliva reduction, Wry Neck in cardinal symptom, late period.At present, for carcinoma of mouth master Operative treatment, radiotherapy, the Compositive sequence in treatment based on chemotherapy are used, but therapeutic effect is often bad, 5 years Survival rate is less than 50%.Preferable anticancer drug should possess low toxicity, safely, effectively, the conditions such as inexpensive, therefore there is an urgent need for research and development.

A kind of new drug both expensive is developed, the cycle is long, is to find having for new anti-cancer drug object using existing known compound Effect means.Fluoxastrobin (alias Amici reaches, English name Azoxystrobin) is a kind of to be separated from mushroom Strobilurin class natural insecticides have the characteristics that wide spectrum, efficient, low toxicity, protection, treatment.Fluoxastrobin is a kind of mitochondria Respiration inhibitor, molecular formula C₂₂H₁₇N₃O₅, molecular weight 403.3875, CAS 131860-3-8, synthesis patent is earliest by Britain Supreme Being Chemical industrial company of state applies.It is shifted by inhibiting Cytochrome B cl to the electronics of cromoci, so as to inhibit mitochondria Breathing.Fluoxastrobin toxicity is relatively low, and the acute oral, percutaneous toxicity to mouse female, great and mighty or powerful belong to low toxicity, mouse LD50 of passing through mouth female, great and mighty or powerful> 5000mg/kg, acute percutaneous LD50>5000mg/kg, Acute inhalation LC50 (2h)>2474mg/m³, belong to hypotoxicity, priming experiments Draw to guinea pig skin sensitization rate to be 0%.Teratology testing the results show Fluoxastrobin active compound does not cause people periphery to learn lymphocyte dye Colour solid aberration rate increases, not yet reported to have mutagenesis, teratogenesis and carcinogenesis.It is special in the in vivo Drug Pharmacokinetics of rat Point is that 1h blood concentrations reach peak value after Fluoxastrobin administration, have a very wide distribution, but do not have accumulation to make in tested tissue organ With excretion is fast, is mainly directly discharged after 4h with the prototype without metabolism with excrement.

Prior art background defect：Chemotherapy is one of carcinoma of mouth clinical treatment common method, is played in carcinoma of mouth treatment Important role, but there is chemotherapeutics drug resistance phenomenon in some cases body after chemotherapy, chemotherapy is caused to fail, and it is secondary It acts on larger.It is of great significance so finding the efficient medicine of novel low-toxicity to carcinoma of mouth complex treatment.Institute of the present invention The compound Fluoxastrobin being related to is common commercially available wide-spectrum bactericide, is mainly used for the prevention and control of plant diseases, pest control of crops, preparation side Method is simple, good disinsection effect.We have discovered that the growth of Fluoxastrobin oral cancer cell has significant inhibitory action, be expected to for The new drug development of Clinical mouth cancer treatment provides scientific basis.So far, both at home and abroad it is not yet found that pass Fluoxastrobin is related to cancer Journal of Sex Research.

The content of the invention

The defects of the purpose of the present invention is being directed to current carcinoma of mouth complex treatment, provides a kind of preparing as antineoplastic The compound applied in object provides new support for the clinical drug therapy of carcinoma of mouth.

To achieve the above object, the technical solution adopted by the present invention is as follows.

Specifically include following steps：

1. Fluoxastrobin is the compound of structural formula shown in formula (1), molecular formula C₂₂H₁₇N₃O₅, chemical name, (E)-[2- [6- (2- cyano-benzene oxygens) pyrimidine-4-yl oxygen] phenyl] -3- methoxy-methyl acrylates, CAS numbers are 131860-33-8；Appearance For white powder, molecular weight 403.34.This product also dissolves in other organic solvents such as methanol, toluene, acetone, but not as It limits, Fluoxastrobin active compound is dissolved in DMSO in the present invention, be configured to 50mg/ml storage liquid, be stored in 4 DEG C of refrigerators.

2. the influence of Fluoxastrobin oral cancer cell multiplication

Influence of the Fluoxastrobin to tumor cell proliferation is detected by CCK-8 methods, principle is existing for electronics coupled agent In the case of, the orange-yellow formazan product for generating high water soluble can be reduced by Intramitochondrial dehydrogenase.The depth of color with The multiplication of cell is directly proportional, OD values is measured at 450nM wavelength using microplate reader, to reflect living cells quantity indirectly.

The Fluoxastrobin of 1.5 μ g/ml, 3 μ g/ml, 6 μ g/ml, 12 μ g/ml various concentrations is respectively acting on cancer cell of oral cavity After SCC15, CAL2724h, 48h, 72h, cell Proliferation is inhibited be subject to different degrees of.With the rise of drug concentration, SCC15 And CAL27 cell proliferation inhibition rates are also with rise.Fluoxastrobin acts on SCC15 cells for 24 hours, 48h, the half inhibiting rate of 72h IC50 is respectively 7.82 μ g/ml, 7.51 μ g/ml and 8.02 μ g/ml；For 24 hours, the IC50 of 48h, 72h are respectively 4.4 μ to CAL27 cells G/ml, 6.5 μ g/ml and 5.9 μ g/ml.Prompt Fluoxastrobin oral cancer cell multiplication inhibited.

3. the influence that Fluoxastrobin is proliferated oral cavity intraepithelial neoplasia cells

The Fluoxastrobin of 1.5 μ g/ml, 3 μ g/ml, 6 μ g/ml, 12 μ g/ml various concentrations is respectively acting on oral precancerous lesions After becoming cell DOK24h, 48h, 72h, cell Proliferation is significantly inhibited.The inhibiting rate of 48h, 72h all 90%-100% it Between.Prompt Fluoxastrobin especially notable to the inhibited proliferation of oral cavity intraepithelial neoplasia cells.

In conclusion the present invention has expanded the new application approach of Fluoxastrobin, to be applied to Fluoxastrobin to inhibit tumour cell In growth, the therapeutic effect of carcinoma of mouth can be improved further using being included in the medical components for the treatment of carcinoma of mouth.This hair The scope of the bright application to the compound includes the growth inhibition for cancer cell of oral cavity, and then inhibits the hyperplasia of tumour, so as to Delay the deterioration of tumour.Meanwhile Fluoxastrobin can be also used in the medical component for inhibiting cancer cell of oral cavity growth in the present invention Ingredient in, may also include the carrier of pharmaceutical acceptable.Carrier can be excipient (such as water), filler (such as sucrose or shallow lake Powder), binder (such as cellulose derivative), diluent, disintegrant, sorbefacient or sweetener, but be not only limitted to this.This Invention pharmaceutical preparation can be manufactured according to the preparation method of general known pharmacy, and Fluoxastrobin and more than one carrier are mutually mixed It closes, this dosage form may include pastille, pulvis, granula, capsule or other liquid preparations, but not as limit.

Below in conjunction with the embodiment that further illustrates the present invention of diagram,

Description of the drawings

Fig. 1 Fluoxastrobin chemical structural formulas；

Influence of Fig. 2 Fluoxastrobins to SCC15 cell Proliferations；

Influence of Fig. 3 Fluoxastrobins to CAL27 cell Proliferations；

Influence of Fig. 4 Fluoxastrobins to DOK cell Proliferations；

Specific embodiment

In DMEM-F12 culture mediums, CAL27 cells and DOK cell culture are cultivated in DMEM high sugar for SCC15 cell culture In base, l0%Gibco hyclones, 100U/ml penicillin, 100U/ml streptomysins are placed in 5%CO₂, 95% air, 37 DEG C Saturated humidity incubator in cultivate.Cell attachment is grown, and once, cell is in exponential phase needed for experiment for passage in 2-3 days.

Cell Proliferation is detected using CCK-8 methods.Experimental setup experimental group, Fluoxastrobin concentration are respectively 1.5 μ g/ml, 3 μ g/ ml、6μg/ml、12μg/ml；Solvent control group, with experimental group with the DMSO of concentration.It takes the logarithm growth period cell, with containing 0.25% EDTA pancreatin digests, and supernatant is abandoned in centrifugation.It is 2.5 × 10 with the culture medium adjustment cell concentration containing 10% serum⁴A/ml.Often Hole is inoculated with 200 μ l cell suspensions and enters in 96 well culture plates, every group of 4 multiple holes.It is adherent overnight after, suction abandon supernatant, contain by a group addition There is the culture solution (serum content 10%) of various concentration Radix Rhapontici seu Radix Echinopsis EA, and set solvent control group and continue to cultivate different time (for 24 hours, 48h、72h).Add 190 μ l culture mediums and 10 μ l CCK8 reagents per hole, be placed in 37 DEG C, the interior culture of 5%CO2 saturated humidities incubator Culture is terminated after 2.5h.Each hole absorbance (A values) is measured in microplate reader, selects 450nm wavelength, record is as a result, and by following Formula calculates proliferation inhibition rate：Proliferation inhibition rate (%)=(A_{Experimental group}-A_{Blank group})/(A_{Control group}-A_{Blank group}) × 100%.

Claims (3)

1. the application of Fluoxastrobin or its preparation in the drug for preparing prevention carcinoma of mouth, it is characterised in that：The preparation includes Imitate the Fluoxastrobin of dosage and the carrier of pharmaceutical acceptable.

2. application according to claim 1, it is characterised in that：The carrier is diluent.

3. application according to claim 1 or 2, it is characterised in that：The preparation is tablet, dragee, capsule, injection Agent, solution, lotion, ointment, emulsifiable paste, inhalant, aerosol or suppository.