CN109793726A - Benzethonium chloride is preparing the application in the drug for preventing and treating lung cancer - Google Patents

Benzethonium chloride is preparing the application in the drug for preventing and treating lung cancer Download PDF

Info

Publication number
CN109793726A
CN109793726A CN201910189607.1A CN201910189607A CN109793726A CN 109793726 A CN109793726 A CN 109793726A CN 201910189607 A CN201910189607 A CN 201910189607A CN 109793726 A CN109793726 A CN 109793726A
Authority
CN
China
Prior art keywords
benzethonium chloride
lung cancer
drug
gefitinib
application
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
CN201910189607.1A
Other languages
Chinese (zh)
Other versions
CN109793726B (en
Inventor
何庆瑜
李斌
黄小惠
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Jinan University
Original Assignee
Jinan University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Jinan University filed Critical Jinan University
Priority to CN201910189607.1A priority Critical patent/CN109793726B/en
Publication of CN109793726A publication Critical patent/CN109793726A/en
Application granted granted Critical
Publication of CN109793726B publication Critical patent/CN109793726B/en
Expired - Fee Related legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Landscapes

  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

The invention discloses benzethonium chlorides to prepare the application in the drug for preventing and treating lung cancer.The outer experiment in vivo discovery benzethonium chloride of combination can inhibit the growth of lung cancer, proliferation in the present invention, since benzethonium chloride poison has Small side effects, highly-safe, cheap advantage, therefore can be as a kind of drug for treating lung cancer.Furthermore, it has also been found that when benzyl rope chlorine ammonia and Gefitinib are used in combination, the growth of lung carcinoma cell is significantly reduced than exclusive use Gefitinib, illustrate that the addition of benzethonium chloride can increase lung carcinoma cell to the sensibility of Gefitinib, therefore, can be using benzyl rope chlorine ammonia as the sensitizer of Gefitinib, or by benzyl rope chlorine ammonia and Gefitinib drug combination, it is used for lung cancer therapy aspect.

Description

Benzethonium chloride is preparing the application in the drug for preventing and treating lung cancer
Technical field
The invention belongs to pharmaceutical technology field, in particular to benzethonium chloride is preparing answering in the drug for preventing and treating lung cancer With.
Background technique
Lung cancer is one of most common malignant tumour, and one of most refractory disease in the world, in recent years disease incidence Raising trend is presented.Since lung cancer and lung benign disease identify more difficulty, and lung cancer initial symptoms are unobvious, and most of patients is just The state of an illness has developed to middle and advanced stage when examining, thus often misses its optimal treatment period, seriously affects its therapeutic effect.Lung cancer at present Treatment means include operative treatment, radiotherapy, chemotherapy, immunization therapy, traditional Chinese medical herbal treatment etc., however cancer is to putting The tolerance for the treatment of results in patient's postoperative recurrence and poor prognosis, and treatment difficulty increases, and clinically used anticancer drug is past It is past with certain toxic side effect and expensive.Therefore, the safe and efficient anti-tumor drug for inhibiting lung cancer growth is found to carve Do not allow to delay.
Benzethonium chloride (Benzethonium chloride) is food and medicine surveillance authority, the U.S. (FDA) approval for first aid Articles are a kind of disinfection sanitizer and novel preservative, have preferable surface-active, be widely used in daily use chemicals, medicine etc. Do anticorrosion and bactericidal agent in field.But for lung cancer, it is raw to lung carcinoma cell that there is no pertinent literatures and patent report benzethonium chloride With inhibiting effect.
Summary of the invention
The primary purpose of the present invention is that the shortcomings that overcoming the prior art and deficiency, provide benzethonium chloride in preparation for preventing Control the application in the drug of lung cancer.
Another object of the present invention is to provide benzethonium chlorides and Gefitinib in preparing antineoplastic combination drug Using.
The purpose of the invention is achieved by the following technical solution: benzethonium chloride is in preparing the drug for preventing and treating lung cancer Using.
The benzethonium chloride achievees the purpose that treat lung cancer by the growth and/or proliferation that inhibit lung cancer.
The effective concentration of the benzethonium chloride is 5~20 μM;Preferably 10~20 μM.
Benzethonium chloride inhibits the application in the growth of lung carcinoma cell and/or the drug of proliferation in preparation.
The effective concentration of the benzethonium chloride is 5~20 μM;Preferably 10~20 μM.
The lung carcinoma cell is preferably A549 cell or H1299 cell.
Benzethonium chloride and Gefitinib are preparing the application in antineoplastic combination drug.
The tumour is preferably lung cancer.
A kind of antineoplastic combined medicament, including benzethonium chloride and Gefitinib.
The effective concentration of benzethonium chloride is 5~20 μM (preferably 5 μM) in the antineoplastic combined medicament, Gefitinib Effective concentration be 5~20 μM (preferably 5 μM).
Benzethonium chloride increases the application in drug of the lung carcinoma cell to the sensibility of Gefitinib in preparation.
Can also pharmaceutically may be used containing one or more in the drug and antineoplastic combined medicament of the prevention and treatment lung cancer The carrier or auxiliary material of receiving;The auxiliary material is sustained release agent, excipient, filler, adhesive, wetting agent, disintegrating agent, absorbs rush Into at least one of agent, surfactant or lubricant.
The drug can be prepared into pharmaceutical preparation using the conventional method of this field;Drug system including oral administration Agent, such as capsule, pill, tablet, oral solution, granule, tincture.
Application of the benzethonium chloride in the sensitizer for preparing Gefitinib.
The present invention has the following advantages and effects with respect to the prior art:
1, benzethonium chloride is mainly used for disinfection sanitizer, but there is presently no relevant reports for its inhibiting effect to lung cancer And patent.The outer experiment in vivo of combination of the present invention, the results showed that benzethonium chloride can inhibit the growth of lung cancer, proliferation, benzyl rope chlorine Ammonium is FDA approval drug, and toxic side effect is small, highly-safe, cheap, benzethonium chloride can be dissolved with water, is administered orally, Lung cancer therapy is assisted, a kind of new medicament selection can be provided for the clinical treatment of lung cancer, the prospect of development and utilization is good.
2, for Gefitinib as lung cancer fiest-tire medication, generate drug resistance seriously affects therapeutic effect, and in the present invention It was found that benzyl rope chlorine ammonia can increase lung carcinoma cell to the sensibility of Gefitinib, therefore, benzethonium chloride and Gefitinib can be joined Medicine is shared, in terms of lung cancer therapy.
Detailed description of the invention
Fig. 1 is influence diagram of the benzethonium chloride to proliferation of lung cancer cells;Wherein, figure A is the benzethonium chloride pair of various concentration A549 Cell growth ability influences;Scheming the benzethonium chloride that B is various concentration influences the growth ability of H1299 cell.
Fig. 2 is influence diagram of the benzethonium chloride to Increase Apoptosis of Lung Cancer Cells;Wherein, figure A is the Bian rope oronain pair of various concentration The influence of A549 and H1299 Apoptosis;Scheme the statistics knot that the Bian rope oronain that B is various concentration induces A549 apoptosis Fruit;Scheme the statistical result that the Bian rope oronain that C is various concentration induces H1299 apoptosis.
Fig. 3 is influence diagram of the benzethonium chloride to the one-tenth knurl ability of lung carcinoma cell in nude mouse;Wherein, figure A is in nude mouse Subcutaneous tumor;Scheme the variation that B is gross tumor volume size;Scheme the changes of weight that C is nude mice.
Fig. 4 is that benzethonium chloride increases lung carcinoma cell to the result figure of gefitinib-sensitive;Wherein, figure A is different pharmaceutical A549 Cell growth ability is influenced;Scheming B is that different pharmaceutical influences the growth ability of H1299 cell.
Specific embodiment
Below with reference to embodiment, the present invention is described in further detail, and embodiments of the present invention are not limited thereto. Unless stated otherwise, the present invention uses reagent, method and apparatus is the art conventional reagents, method and apparatus.Unless It illustrates, agents useful for same and raw material of the present invention can pass through commercially available acquisition.
Embodiment 1WST-1 cytoactive detection
1. plating cells: collecting A549 and H1299 cell, (purchase is in American Type Culture Collecti (AmericanType Culture Collection, ATCC) cell bank), cell count is carried out, is laid on 96 orifice plates according to the amount of every 3000 cells in hole In, overnight incubation.
2. drug-treated: diluting benzethonium chloride to concentration for the treatment of (0,5,10,15,20 μM), so with DMEM complete medium After replace original culture medium in 96 orifice plates, respectively handle 24,48 and 72h.
3.WST-1 detection: according to the dilution ratio of 1:10 (w/v), with DMEM complete medium dilution WST-1, (WST-1 is thin Born of the same parents are proliferated detection kit), obtain the culture medium containing WST-1;Then 96 orifice plate Central Plains are replaced with the culture medium containing WST-1 In addition culture medium containing WST-1 is added in the hole without culture cell and is used as blank control, every 100 μ of hole by some culture mediums L.96 orifice plates are put in again after being incubated for 1h in 37 DEG C of constant incubators, detect (450nm) with microplate reader read plate.Whole process is protected from light behaviour Make.
4. result: result is as shown in Figure 1A and 1B, shows in figure, shows at the drug of concentration gradient and time gradient Reason, the growth of lung carcinoma cell receive apparent inhibition, i.e. the benzethonium chloride proliferation that can be used for inhibiting lung carcinoma cell.
2 Apoptosis of embodiment
1. drug-treated: collect A549 and H1299 cell be laid in 6 orifice plates, making cell density is about 40%, respectively plus Enter the DMEM medium treatment cell containing 0,10,15 and 20 μM of benzethonium chloride.
2. collecting cell: after drug-treated 48h, the cell that apoptosis occurs in supernatant is first collected, after 1100rpm is centrifuged 3min It abandons supernatant to be collected in same centrifuge tube then 0.25% trypsin digestion of adherent cell, 1100rpm centrifugation Supernatant is abandoned after 3min.PBS buffer solution cleaning is dyed afterwards twice.
3. dyeing: cell is resuspended with the Binding Buffer (cell apoptosis detection kit) of 500 μ L first, then often The Annexin V-FITC that 5 μ L are added in pipe adds 5 μ L Propidium Iodide after mixing is mixed, and it is anti-to be then protected from light room temperature Answer 15min.
4. flow cytometer detection: using C6 flow cytomery after dyeing, before upper machine, sample is filtered off with 400 mesh filter screens Except particulate matter.Wherein Annexin V-FITC is green fluorescence, selects FL1 Air conduct measurement, and Propidium Iodide is red Fluorescence selects FL2 FL3 Air conduct measurement.
5. result: result as indicated with 2, Fig. 2 show benzethonium chloride processing after, it will be apparent that induction of the apoptosis of cell.
3 nude mice by subcutaneous tumor formation of embodiment experiment
1. the zoopery of project strictly observes the guideline of animal care mechanism, and obtains Ji'nan University animal The approval of the Experimental Ethical committee.
2. experimental animal used in project is that (Balb/c-nu is bought big in southern medical courses in general 6~8 week old female nude mices Learn Experimental Animal Center), totally 15, wherein control group 5 and experimental group 10 construct nude mice by subcutaneous tumor formation model:
(1) cell is collected: being collected cell and is carried out cell count, with every nude mouse 1 × 106The amount of a A549 cell calculates Total cell number, cell (are bought in Guangzhou Si Er Biotechnology Co., Ltd, coring) after being resuspended with PBS buffer solution with matrigel 1:1 is mixed by volume.
(2) it is inoculated with: mouse being anaesthetized and (by painless and have a pain stimulation and assess anesthesia level, determines that nude mice is in Narcosis) after, take the above-mentioned and mixed resuspension cell of matrigel to carry out subcutaneously nude mice with 1mL syringe (25G syringe needle) Injection.
3. cell infusion after a week, is observed the tumor formation situation of nude mouse and measured, when tumour reaches 5 × 5 size Afterwards, start by be injected intraperitoneally and stomach-filling in a manner of be administered.3 benzethonium chloride concentration gradients are arranged altogether and (first use DMSO for this experiment Benzethonium chloride mother liquor 50mg/mL is prepared, then is diluted to sterile water using concentration, 100 μ L are administered in every mouse every time), two kinds Administration mode.Respectively 0mg/kg, 5mg/kg (intraperitoneal injection) and 10mg/kg (stomach-filling), are administered once for every two days by each 5.6. In administration process, the size of nude mice weight and transplantable tumor was measured in every two days.After two weeks, nude mice is euthanized and by tumour It takes out.
4. result: result is as shown in Figure 3.Fig. 3 A and 3B show the subendothelial tumor formation experimental result of nude mouse, the growth of tumour Curve shows obvious inhibition of the subendothelial tumor formation of nude mouse by benzethonium chloride.Show that benzethonium chloride can inhibit the tumour of lung cancer Growth.And Fig. 3 C is the weight statistical result of three groups of mouse after administration, shows that weight has no significant difference between three groups of mouse, Prove the safety that uses of the benzethonium chloride as anti-tumor drug.
4 benzyl rope chlorine ammonia of embodiment increases lung carcinoma cell to the sensibility of Gefitinib
1. plating cells: collecting A549 and H1299 cell, carry out cell count, spread according to the amount of 3000, every hole cell In 96 orifice plates, overnight incubation.
2. drug-treated: then replacing 96 to 5 μM with DMEM complete medium dilution benzethonium chloride and Gefitinib respectively Original culture medium in orifice plate handles 72h.
3.WST-1 detection: according to the dilution ratio of 1:10 (w/v), with DMEM complete medium dilution WST-1, (WST-1 is thin Born of the same parents are proliferated detection kit), obtain the culture medium containing WST-1;Then 96 orifice plate Central Plains are replaced with the culture medium containing WST-1 In addition culture medium containing WST-1 is added in the hole without culture cell and is used as blank control, every 100 μ of hole by some culture mediums L.96 orifice plates are put in again after being incubated for 1h in 37 DEG C of constant incubators, detect (450nm) with microplate reader read plate.Whole process is protected from light behaviour Make.
4. result: as shown in figure 4, Gefitinib group is used alone compared with group is used in combination, it will be apparent that inhibit lung cancer The growth of cell illustrates that the addition of benzethonium chloride can increase lung carcinoma cell to the sensitivity of Gefitinib there are significant difference Property.
The above embodiment is a preferred embodiment of the present invention, but embodiments of the present invention are not by above-described embodiment Limitation, other any changes, modifications, substitutions, combinations, simplifications made without departing from the spirit and principles of the present invention, It should be equivalent substitute mode, be included within the scope of the present invention.

Claims (10)

1. benzethonium chloride is preparing the application in the drug for preventing and treating lung cancer.
2. benzethonium chloride according to claim 1 is preparing the application in the drug for preventing and treating lung cancer, it is characterised in that: The benzethonium chloride achievees the purpose that treat lung cancer by the growth and/or proliferation that inhibit lung cancer.
3. benzethonium chloride according to claim 1 is preparing the application in the drug for preventing and treating lung cancer, it is characterised in that: The effective concentration of the benzethonium chloride is 5~20 μM.
4. benzethonium chloride inhibits the application in the growth of lung carcinoma cell and/or the drug of proliferation in preparation.
5. benzethonium chloride and Gefitinib are preparing the application in antineoplastic combination drug.
6. benzethonium chloride according to claim 5 and Gefitinib are preparing the application in antineoplastic combination drug, Be characterized in that: the tumour is lung cancer.
7. a kind of antineoplastic combined medicament, it is characterised in that: including benzethonium chloride and Gefitinib.
8. antineoplastic combined medicament according to claim 7, it is characterised in that: benzyl rope in the antineoplastic combined medicament The effective concentration of oronain is 5~20 μM, and the effective concentration of Gefitinib is 5~20 μM.
9. benzethonium chloride increases the application in drug of the lung carcinoma cell to the sensibility of Gefitinib in preparation.
10. application of the benzethonium chloride in the sensitizer for preparing Gefitinib.
CN201910189607.1A 2019-03-13 2019-03-13 Application of benzethonium chloride in preparation of medicine for preventing and treating lung cancer Expired - Fee Related CN109793726B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201910189607.1A CN109793726B (en) 2019-03-13 2019-03-13 Application of benzethonium chloride in preparation of medicine for preventing and treating lung cancer

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201910189607.1A CN109793726B (en) 2019-03-13 2019-03-13 Application of benzethonium chloride in preparation of medicine for preventing and treating lung cancer

Publications (2)

Publication Number Publication Date
CN109793726A true CN109793726A (en) 2019-05-24
CN109793726B CN109793726B (en) 2021-02-05

Family

ID=66563544

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201910189607.1A Expired - Fee Related CN109793726B (en) 2019-03-13 2019-03-13 Application of benzethonium chloride in preparation of medicine for preventing and treating lung cancer

Country Status (1)

Country Link
CN (1) CN109793726B (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115634219A (en) * 2022-09-13 2023-01-24 武汉科技大学 Application of benzethonium chloride in preparation of medicine for preventing and treating triple negative breast cancer

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2008122038A1 (en) * 2007-04-02 2008-10-09 President And Fellows Of Harvard College Regulating autophagy

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2008122038A1 (en) * 2007-04-02 2008-10-09 President And Fellows Of Harvard College Regulating autophagy

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115634219A (en) * 2022-09-13 2023-01-24 武汉科技大学 Application of benzethonium chloride in preparation of medicine for preventing and treating triple negative breast cancer

Also Published As

Publication number Publication date
CN109793726B (en) 2021-02-05

Similar Documents

Publication Publication Date Title
CN109793726A (en) Benzethonium chloride is preparing the application in the drug for preventing and treating lung cancer
CN110755431B (en) Application of desloratadine in preparation of anti-liver cancer drugs
CN104707097B (en) Pharmaceutical composition Mongolian oak stilbene dissipates and its application in the drug for blocking hepatic precancerosis, treatment liver cancer or virus hepatitis is prepared
CN101125140B (en) Application of dihydroartemisinin in reinforcing chemotherapy medicine antitumor curative effect
CN109793738A (en) Daurisoline is preparing the application in the drug for preventing and treating lung cancer
CN102727495A (en) Novel use of sanguinarine chloride
CN108853114B (en) Application of nifurolimus in preparation of medicine for treating cancer-derived brain metastasis tumor
CN102058854B (en) Chinese medicine for treating malignant lymphoma
CN110179859A (en) The application of Radix Stephaniae Tetrandrae extract and fangchinoline in pharmacy
CN113425721A (en) Application of lomitapide in preparation of antitumor drugs
CN100546621C (en) A kind of new purposes of known drug
CN100502843C (en) Chinese medicine for treating mammary gland cystoma, mammitis and preparation method of its film coated tablet
CN103251876A (en) Traditional Chinese medicine composition used for treating chronic liver diseases
CN110354121A (en) Application of the Ciclopirox Olamine in preparation tumor and composition of medicine and purposes comprising Ciclopirox Olamine
CN110339195A (en) For treating the quinoline of cholangiocarcinoma
CN110051734A (en) A kind of pharmaceutical composition and its application of resistive connection enteritis correlation colon cancer
CN114224989B (en) Traditional Chinese medicine composition for resisting helicobacter pylori
CN115252613B (en) Pharmaceutical composition and application thereof in reversing drug resistance of lenvatinib
CN116270489B (en) Ginger stomach-activating granule and preparation method thereof
CN116407600B (en) Traditional Chinese medicine compound for treating lung cancer and application of traditional Chinese medicine compound for treating lung nodule
CN115634219A (en) Application of benzethonium chloride in preparation of medicine for preventing and treating triple negative breast cancer
CN108498570A (en) A kind of Chinese medicine compound prescription and preparation method thereof for anti-idiopathic pulmonary fibrosis
CN101797242A (en) Application of cysteamine in preparing medicine for treating cancer
CN101708271B (en) Chinese medicament for treating esophageal cancer
CN102068588B (en) Chinese medicinal preparation for treating hepatopathy and preparation method thereof

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant
CF01 Termination of patent right due to non-payment of annual fee
CF01 Termination of patent right due to non-payment of annual fee

Granted publication date: 20210205