CN110507651A

CN110507651A - A kind of drug and application thereof with anti-non-small cell lung cancer effect

Info

Publication number: CN110507651A
Application number: CN201910620036.2A
Authority: CN
Inventors: 陈美姿; 周融融; 胡洪波; 彭咏波
Original assignee: CHENZHOU FIRST PEOPLE'S HOSPITAL
Current assignee: CHENZHOU FIRST PEOPLE'S HOSPITAL
Priority date: 2019-07-10
Filing date: 2019-07-10
Publication date: 2019-11-29
Anticipated expiration: 2039-07-10
Also published as: CN110507651B

Abstract

The invention belongs to drug therapy fields, disclose a kind of drug and application thereof with anti-non-small cell lung cancer effect.The active constituent of the drug is Febustat.The method for being used to treat non-small cell lung cancer present invention firstly provides Febustat and its application in preparation treatment non-small cell lung cancer drug, it will be a very promising use of approved drugs for nonapproved uses clinical strategy that Febustat, which is applied to treatment non-small cell lung cancer, benefit more Patients with Non-small-cell Lung.

Description

A kind of drug and application thereof with anti-non-small cell lung cancer effect

Technical field

The invention belongs to drug therapy field, in particular to a kind of drug with anti-non-small cell lung cancer effect and its Purposes.

Background technique

Cancer is to seriously endanger a big chronic disease of human health, has become the second largest killer for being only second to cardiovascular disease. Lung cancer is first of the highest malignant tumour of morbidity and mortality, wherein non-small cell lung cancer (Non-small cell lung Cancer, abbreviation NSCLC) account for the 80% of lung cancer.Currently, the primary treatments of non-small cell lung cancer are still chemotherapy.For There are EGFR^L858RThere are the patient of different deletion mutations (common Asia, women, non-suctions for mutation or EGFR19 exon Cigarette, Patients with Non-small-cell Lung) apply Gefitinib (Gefitinib, Gef；Also known as Iressa) targeted therapy effect is preferable.

The biggish puzzlement of the generally existing side effect of chemotherapy of tumors, because it is while killing tumour cell also to normal Cell has stronger lethal effect, and many tumor patients can not be resistant to the chemotherapy of larger dose and cause treatment that can not continue And it is dead.Although targeted drug Gefitinib has certain therapeutic effect to the part EGFR patient that there is mutation, in clinic On also achieve significant curative effect, but using after several courses for the treatment of, can also generate certain drug resistance and side effect, patient is general It can occur to recur in 1~2 year or shift, or generate new mutation and drug resistance is generated to Gefitinib, so, even if making It still cannot effectively be improved 5 years of Patients with Non-small-cell Lung with Gefitinib (tyrosine kinase inhibitor) treatment Overall survival.Some researches show that certain Chinese traditional medicine molecules can reduce the drug resistance that anticancer drug generates in use and secondary work With so that improving it treats effectiveness.

Febustat (Febuxostat, abbreviation FBS), Fei Buzuosita, entitled the 2- [(3- cyano -4- isobutyl of chemistry Oxygroup) phenyl] -4- methyl-5-thiazole carboxylic acid is xanthine oxidase (XO) inhibitor, suitable for the height with gout symptom The long-term treatment of uricacidemia, clinical application are significantly safe.

Summary of the invention

In order to overcome shortcoming and defect existing in the prior art, the primary purpose of the present invention is that providing one kind has The drug of anti-non-small cell lung cancer effect.

Another object of the present invention is to provide a kind of purposes of said medicine.

The purpose of the invention is achieved by the following technical solution:

A kind of drug with anti-non-small cell lung cancer effect, the active constituent of the drug are Febustat.

The drug also contains pharmaceutically acceptable carrier.

The drug is prepared by conventional fabrication process.

Above-mentioned drug is preparing the purposes in anti-non-small cell lung cancer drug.

The drug can be used for the drug therapy of mammal including people.

The present invention have the advantages that compared with prior art as follows protrude and the utility model has the advantages that

Inventor is respectively to three kinds of different types of non-small cell lung cancer cell strain NCI-H1975 (EGFR^L858R/T790M), HCC827(EGFR^E746-A750del) and A549 (EGFR^WT) it is that model carries out external MTT screening, find it with very strong anti- Proliferation activity and be in good timeliness and dose-effect relationship, has good clinical application development prospect again.Meanwhile etc. effects Dose evaluation shows that combination drug not yet increases the toxicity of normal liver cell L-O2.

Detailed description of the invention

Fig. 1 is figure compared with Febustat is acted on altogether with Gefitinib to the degradation of EGFR protein expression.

Fig. 2 is that Febustat and Gefitinib are applied alone and are combined and inhibit situation map to H1975 mdr cell transplantable tumor.

Fig. 3 is that Febustat and Gefitinib are applied alone and are combined to H1975 cell transplantation tumor nude mice weight change figure.

Specific embodiment

Present invention will now be described in further detail with reference to the embodiments and the accompanying drawings, but embodiments of the present invention are not It is limited to this.

Embodiment 1:NCI-H1975 (EGFR^L858R/T790M), HCC827 (EGFR^E746-A750del) and A549 (EGFR^WT) Cell screening optimizes Gefitinib Gef and Febustat FBS composition

The cell of logarithmic growth phase is inoculated with 3 × 10 respectively⁴A cells/well is on 96 orifice plates, after 6 hours to be grown, Centrifugation abandon supernatant, then by following grouping administration: tumour cell sets not dosing group and dosing group, wherein dosing group set Gef with The mono- medicine group of FBS, Gef and FBS drug combination different mol ratio example group, every group sets 4-6 multiple holes, cultivates 24 hours, abandons supernatant, MTT (tetrazolium) serum-free medium of the 100 μ l containing 0.5mg/ml is added culture 4 hours, 100 μ l DMSO (diformazans are added Sulfoxide), it is placed on micro-oscillating instrument and vibrates 10min, then be placed in microplate reader detection OD value at 570nm.As a result according to following Inhibiting rate formula calculates the inhibiting rate of growth of tumour cell in each case, and concrete outcome is shown in Table 1.

Inhibiting rate=(1- dosing group OD value/control group OD value)

Table 1 is that FBS acts on IC of three kinds of different lung carcinoma cells after 72 hours₅₀Inhibit situation.We can be found that with dense The increase of degree, inhibiting rate are in significant concentration dependent, wherein discovery Febustat cell H1975 drug resistant to Gefitinib It is same sensitive, further FBS drug is prompted to all have significant response to treatment to three groups of non-small cell lung cancer cell strains.

1 FBS of table acts on 72 hours IC₅₀(μM)

The expression of embodiment 2:EGFR and Western blot detection

Immune Western blot antibody, buys in the C-Caspase-3 and anti-EGFR of Abcam company (ab52894)(Abcam).After three kinds of cells apply Gefitinib Gef or Febustat FBS to handle 48 hours respectively, 5 are collected ×10⁶Cell applies RIPA (50mM Tris pH 8.0, the 150mM NaCl, 0.1%SDS, 0.5%sodium of 200 μ l Deoxycholate, 1%NP-40) addition protease inhibitors crack 30 minutes on ice after, 12000rpm be centrifuged 10 minutes, receive Collect supernatant, BCA method measures protein concentration, and 2 × SDS loading buffer is added and boils in 100 DEG C 10 minutes.Total protein (100 μ g) loading according to the molecular size range application 8-15%SDS-PAGE gel electrophoresis of albumen is then transferred on pvdf membrane (GE Healthcare), corresponding antibody incubation is applied respectively, and (Millipore is public by chemiluminescence zymolyte HRP Substrate Department), using LAS-4000 imaging system images (Fuji).Western blot experimental result as shown in Figure 1, we in albumen Level has detected the expression of EGFR albumen.To three kinds of different non-small cell lung cancer cells, FBS drug ratio Gef group is all The EGFR and C-Caspase-3 of cracking can be more preferably induced to degrade.

The tumour of embodiment 3:FBS Drug inhibition non-small cell lung cancer tumor-bearing mice increases

In order to detect FBS to the curative effect of living body non-small cell lung cancer, we are built in nude mice using NCI-H1975 cell Non-small cell lung cancer mouse tumor model is found.Including control group (Control) (physiological saline)；(300mg/kg/d is filled FBS Stomach) and Gef (25mg/kg/d, stomach-filling) group and with the Gef/FBS combination group that is applied alone dosage to be equal, each group 10, nude mice by subcutaneous After inoculated tumour, when subcutaneous tumor volumes are greater than 100mm³When mouse is randomly divided into four groups, start gastric infusion after the 7th day, Daily gastric infusion is primary, weigh in every other day variation and gross tumor volume, prepares suspension with normal saline dilution.Daily with trip The major diameter and minor axis of calliper to measure tumour are marked, and presses long × wide × wide/2 calculating subcutaneous volumes of its tumour, practical gross tumor volume Change curve such as Fig. 2 (p < 0.01).The tumour initial volume of each group is close.After 13 days, the tumour growth rate of control group is obvious Faster than other three groups, by the 16th day (gastric infusion 9 days), the gross tumor volume of Gef group and FBS group was smaller than control group, and FBS The decline of group is particularly significant (Fig. 2, wherein p < 0.01)；Changes of weight tracking display, FBS medicine group nude mice weight loss are smaller (Fig. 3, wherein ##, p < 0.01) prompts FBS pharmaceutical composition to have extraordinary therapeutic effect and less toxic side effect.

Embodiment 4

Prescription:

Preparation process:

Drug and auxiliary material are crossed into 80 meshes respectively, by 200 grams of Febustat and 48 grams of microcrystalline celluloses and 12 grams of carboxymethyls Sodium starch is sufficiently mixed, 10% starch slurry softwood, the granulation of 18 meshes, dry at 60 DEG C, obtains particle 1.By Febustat 175 It gram is sufficiently mixed with 24 grams of microcrystalline celluloses, 15 grams of starch and 8 grams of sodium carboxymethyl starches, 10% starch slurry softwood, 18 meshes It pelletizes, it is dry at 60 DEG C, obtain particle 2.By equal increments principle, particle 1 and particle 2 are sufficiently mixed, 16 mesh sieves add Enter magnesium stearate, mixes, tabletting, slice weight 500mg.

The embodiments described above only express several embodiments of the present invention, and the description thereof is more specific and detailed, but simultaneously Limitations on the scope of the patent of the present invention therefore cannot be interpreted as.It should be pointed out that for those of ordinary skill in the art For, without departing from the inventive concept of the premise, various modifications and improvements can be made, these belong to of the invention Protection scope.Therefore, the scope of protection of the patent of the invention shall be subject to the appended claims.

Claims

1. a kind of drug with anti-non-small cell lung cancer effect, it is characterised in that: the active constituent of the drug is Febustat.

2. a kind of drug with anti-non-small cell lung cancer effect according to claim 1, it is characterised in that: the drug Also contain pharmaceutically acceptable carrier.

3. drug according to claim 1 is preparing the purposes in anti-non-small cell lung cancer drug.