WO2021023291A1

WO2021023291A1 - Use of proflavine in treatment of lung cancers

Info

Publication number: WO2021023291A1
Application number: PCT/CN2020/107683
Authority: WO
Inventors: 陈毅歆; 王国松; 洪俊平; 吴倩; 陈瑞琪; 黄鹏飞; 夏宁邵
Original assignee: 厦门大学; 养生堂有限公司
Priority date: 2019-08-08
Filing date: 2020-08-07
Publication date: 2021-02-11
Also published as: CN112336864A

Abstract

Disclosed is the use of proflavine, a stereoisomer thereof, a solvate thereof, a pharmaceutically acceptable salt thereof, a hydrate or solvate of the pharmaceutically acceptable salt thereof, or pharmaceutically acceptable modifiers thereof in the preparation of a drug for treating lung cancers. Proflavine can inhibit the proliferation and metastasis of lung cancer cells by means of various mechanisms such as the inhibition of tumor cell proliferation and the inhibition of tumor metastasis, thereby achieving the effect of inhibiting tumor growth or removing tumors.

Description

Application of Proxanthin in the Treatment of Lung Cancer

This application is based on the application with the CN application number 201910728472.1 and the filing date on August 8, 2019, and claims its priority. The disclosure of the CN application is hereby incorporated into this application as a whole.

Technical field

This application relates to the field of medicinal chemistry, specifically to the application of proxanthin in the treatment of lung cancer.

Background technique

Cancer is a serious hazard to human health around the world. According to the "2018 Global Cancer Statistics" report published by the official journal of the American Cancer Society "Clinician Cancer Journal", it is estimated that there will be approximately 18.1 million new cancer cases and 960 late cancer deaths worldwide in 2018 Among them, lung cancer is the world's largest cancer, with morbidity (11.6%) and mortality (18.4%) in the first place. Lung cancer has complex causes and pathogenesis, and tumor cells are highly complex and heterogeneous. Currently, lung cancer is divided into two major types in clinical practice, non-small cell lung cancer and small cell lung cancer. Non-small cell lung cancer can be divided into lung squamous cell carcinoma, lung adenocarcinoma, and large cell lung cancer. Therapeutic drugs for lung cancer have very important prospects, but they will also face huge challenges.

At present, therapeutic drugs for lung cancer are expensive, and patients are often unbearable. Some patients even give up treatment opportunities because they cannot afford the high prices. Therefore, it is of great significance to develop low-cost small-molecule chemicals with good therapeutic effects.

Proflavine, its chemical name is 3,6-diaminoacridine, also known as Proflavine, Proflavine, Proflavine, CAS number: 92-62-6, and the structural formula is as follows:

Proxanthin can be used as a disinfectant, bacteriostatic agent, also used to treat skin pimples, wounds and various inflammations, and used to disinfect skin and wounds. Proxanthin is used in combination with anti-tuberculosis drugs, which has a synergistic effect.

So far, the broad-spectrum therapeutic effect of proxanthin in lung cancer has not been reported.

Application content

The inventor of the present application unexpectedly discovered that the small molecule compound proxanthin sulfate has broad-spectrum therapeutic activity on lung cancer.

This application relates to proxanthin, its stereoisomers, its solvates, its pharmaceutically acceptable salts, the hydrates or solvates of its pharmaceutically acceptable salts, or its pharmaceutically acceptable various Of the modified substance in the preparation of a medicine for treating lung cancer.

The application also relates to the use of a pharmaceutical composition in the preparation of a medicine for treating lung cancer, wherein the pharmaceutical composition contains proxanthin, its stereoisomers, its solvates, its pharmaceutically acceptable salts, and A hydrate or solvate of a pharmaceutically acceptable salt, or various pharmaceutically acceptable modifications thereof, and a pharmaceutically acceptable carrier or excipient.

This application also relates to proxanthin, its stereoisomer, its solvate, its pharmaceutically acceptable salt, the hydrate or solvate of its pharmaceutically acceptable salt, or its pharmaceutically acceptable Various modifications are used in combination with the second anti-tumor drug to prepare drugs for treating lung cancer.

This application also relates to proxanthin, its stereoisomer, its solvate, its pharmaceutically acceptable salt, the hydrate or solvate of its pharmaceutically acceptable salt, or its pharmaceutically acceptable Various modifications, which are used to treat lung cancer.

This application also relates to a pharmaceutical composition for the treatment of lung cancer, wherein the pharmaceutical composition contains proxanthin, its stereoisomers, its solvates, its pharmaceutically acceptable salts, and its pharmaceutically acceptable Salt hydrates or solvates, or various pharmaceutically acceptable modifications thereof, and pharmaceutically acceptable carriers or excipients.

The application also relates to a hydrate or solvate containing proxanthin, its stereoisomers, its solvates, its pharmaceutically acceptable salts, its pharmaceutically acceptable salts, or its pharmaceutically acceptable A combination product of the various modifications of and the second anti-tumor drug, which is used to treat lung cancer.

This application also relates to a method for treating lung cancer, which comprises: administering to a subject in need a therapeutically effective amount of proxanthin, its stereoisomers, its solvates, its pharmaceutically acceptable salts, A hydrate or solvate of its pharmaceutically acceptable salt, or various pharmaceutically acceptable modifications thereof.

This application also relates to another method for treating lung cancer, which comprises: administering a therapeutically effective amount of the pharmaceutical composition described in this application to a subject in need thereof.

This application also relates to another method for treating lung cancer, which comprises: administering a therapeutically effective amount of proxanthin, its stereoisomers, its solvates, and its pharmaceutically acceptable salts to a subject in need , The hydrate or solvate of its pharmaceutically acceptable salt, or its various pharmaceutically acceptable modifications, and the second anti-tumor drug with therapeutic sales.

According to certain embodiments of the present application, the lung cancer described in the present application is selected from: lung squamous cell carcinoma, lung adenocarcinoma, large cell lung cancer, small cell lung cancer, or any combination thereof.

According to certain embodiments of the present application, the pharmaceutical composition described in the present application further contains a second anti-tumor drug.

According to certain embodiments of the present application, the pharmaceutically acceptable salt described in the present application is Proflavine hemisulfate or 3,6-Acridinediamine hydrochloride (Proflavine hydrochloride; Proflavine monohydrochloride) ,

According to certain embodiments of the application, the hydrate of the pharmaceutically acceptable salt described in the application is proflavine hemisulfate hydrate (3,6-Acridinediamine sulfate hydrate (2:1:1); Proflavine hemisulfate hydrate) or Proflavine hydrochloride hemihydrate (3,6-Acridinediamine hydrochloride hydrate (2:2:1); Proflavine hydrochloride hemihydrate; Proflavine monohydrochloride hemihydrate).

According to some embodiments of the present application, the second anti-tumor drug described in the present application is an immune checkpoint inhibitor, a traditional chemotherapeutic drug, or a small molecule inhibitor.

According to certain embodiments of the present application, the immune checkpoint inhibitors described in the present application include but are not limited to: PD-1 inhibitors, PD-L1 inhibitors, CTLA-4 inhibitors, and the like.

According to certain embodiments of the present application, the traditional chemotherapeutic drugs described in the present application include but are not limited to: paclitaxel, cisplatin, 5-fluorouracil and the like.

According to certain embodiments of the present application, the small molecule inhibitors described in the present application include but are not limited to EGFR inhibitors, HER2 inhibitors, ALK inhibitors and the like.

According to certain embodiments of the application, the PD-1 inhibitors described in the application include but are not limited to: Nivolumab, Pembrolizumab, IBI-308, Camrelizumab (SHR-1210), Tislelizumab, Cemiplimab, BCD-100, TSR-042 , PDR-001, JNJ-63723283) etc.

According to certain embodiments of the present application, the PD-L1 inhibitors described in the present application include but are not limited to: Atezolizumab, Avelumab, Durvalumab, BMS-936559, M-7824, CX-072) and the like.

According to certain embodiments of the present application, the CTLA4 inhibitors described in the present application include but are not limited to: Ipilimumab, Tremelimumab, AGEN-1884, AK-104) and the like.

According to certain embodiments of the present application, the EGFR inhibitors described in the present application include but are not limited to: Cetuximab, Erlotinib, Gefitinib, Lapatinib Ditosylate, Neratinib, Theliatinib, Cyasterone, Osimertinib, Avitinib, Afatinib, Gefitinib, Canertinib, Lapatinib, AG -490, Pelitinib, Dacomitinib, etc.

According to certain embodiments of this application, the HER2 inhibitors described in this application include but are not limited to: Poziotinib, Trastuzumab, Pertuzumab, Irbinitinib and the like.

According to certain embodiments of this application, the ALK inhibitors described in this application include but are not limited to: Crizotinib, TAE684 (NVP-TAE684), Alectinib, AZD3463, ASP3026, Brigatinib (AP26113), Ensartinib (X-396), Alectinib Wait.

According to certain embodiments of the present application, the pharmaceutical composition described in the present application may be a tablet, capsule, pill, oral liquid preparation, granule, powder or injection.

According to certain embodiments of the present application, the pharmaceutical composition described in the present application can be administered by oral, injection, implantation, topical application, spraying or inhalation.

According to certain embodiments of the application, the proxanthin of the application, its stereoisomers, its solvates, its pharmaceutically acceptable salts, the hydrates or solvates of its pharmaceutically acceptable salts, or Various pharmaceutically acceptable modifications thereof are used as cytotoxic drugs, tumor growth inhibitors, anti-tumor metastasis drugs and/or anti-tumor invasion drugs when used in the treatment of lung cancer.

According to certain embodiments of the present application, proxanthin, its stereoisomers, its solvates, its pharmaceutically acceptable salts, the hydrates or solvates of its pharmaceutically acceptable salts, or the When various pharmaceutically acceptable modifications are used in combination with the second anti-tumor drug, proxanthin, its stereoisomers, its solvates, its pharmaceutically acceptable salts, and its pharmaceutically acceptable salts Hydrates or solvates, or various pharmaceutically acceptable modifications thereof, are in the same preparation unit as the second anti-tumor drug, or proxanthin, its stereoisomers, its solvates, or its pharmaceutical The above-acceptable salt, the hydrate or solvate of the pharmaceutically acceptable salt, or various pharmaceutically acceptable modifications thereof are combined with the second anti-tumor drug in different preparation units.

According to certain embodiments of the present application, proxanthin, its stereoisomers, its solvates, its pharmaceutically acceptable salts, the hydrates or solvates of its pharmaceutically acceptable salts, or the When various pharmaceutically acceptable modifications are used in combination with the second anti-tumor drug, proxanthin, its stereoisomers, its solvates, its pharmaceutically acceptable salts, and its pharmaceutically acceptable salts Hydrates or solvates, or various pharmaceutically acceptable modifications thereof and the second anti-tumor drug can be administered to the individual in need of treatment at the same time or separately; or proxanthin, its stereoisomers, Its solvate, its pharmaceutically acceptable salt, its pharmaceutically acceptable salt hydrate or solvate, or its various pharmaceutically acceptable modifications, and the second antibody is administered after a certain interval of time Tumor drugs; or administer the second antitumor drug first, and then administer proxanthin, its stereoisomers, its solvates, its pharmaceutically acceptable salts, and the hydration of its pharmaceutically acceptable salts after a certain interval Or solvates, or various pharmaceutically acceptable modifications thereof.

The pharmaceutically acceptable salts of proxanthin described in this application include salts formed by proxanthin and pharmaceutically acceptable inorganic or organic acids. Examples of suitable pharmaceutically acceptable salts include hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, perchloric acid, fumaric acid, acetic acid, propionic acid, succinic acid, glycolic acid, formic acid, lactic acid, maleic acid , Tartaric acid, citric acid, hexanoic acid, malonic acid, hydroxymaleic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, fumaric acid, toluenesulfonic acid, methanesulfonic acid, naphthalene-2-sulfonic acid , Benzenesulfonic acid, hydroxynaphthoic acid, hydroiodic acid, malic acid, tannic acid and other salts.

For example, proflavine sulfate formed by proflavine and sulfuric acid has the English name: Proflavine hemisulfate, CAS number: 1811-28-5, and its molecular structure:

For example, the proflavine hydrochloride formed by proflavine and hydrochloric acid has the English name: Proflavine hydrochloride or Proflavine monohydrochloride, CAS number: 952-23-8, and its molecular structure is:

The hydrate of the pharmaceutically acceptable salt of proxanthin described in this application refers to the hydrate formed by the pharmaceutically acceptable salt of proxanthin described in this application and water. Examples of suitable pharmaceutically acceptable salt hydrates include proflavine hemisulfate hydrate (3,6-Acridinediamine sulfate hydrate (2:1:1); Proflavine hemisulfate hydrate) or proflavine hemisulfate hydrate (3,6-Acridinediamine sulfate hydrate (2:1:1); 6-Acridinediamine hydrochloride hydrate(2:2:1); Proflavine hydrochloride hemihydrate; Proflavine monohydrochloride hemihydrate).

For example, the structural formula of proxanthin sulfate hydrate is as follows:

For example, the structural formula of proxanthin hydrochloride hydrate is as follows:

The carrier described in this application includes, but is not limited to: ion exchangers, alumina, aluminum stearate, lecithin, serum proteins such as human albumin, buffer substances such as phosphate, glycerin, sorbic acid, potassium sorbate, saturated Partial glyceride mixture of plant fatty acids, water, salt or electrolyte, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salt, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, Cellulosic substances, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylate, beeswax, lanolin.

The excipients mentioned in this application refer to additions other than the main drug in the pharmaceutical preparation. It is stable in nature, has no compatibility with the main drug, does not produce side effects, does not affect the curative effect, is not easy to deform, dry, crack, mold, moth, is harmless to the human body, has no physiological effect, and does not produce chemical or physical effects with the main drug It does not affect the content determination of the main drug. Such as binders, fillers, disintegrants, lubricants in tablets; preservatives, antioxidants, flavors, fragrances, cosolvents, emulsifiers, solubilizers, and osmotic pressure regulators in oral liquid preparations , Coloring agents, etc. can all be called excipients, and so on.

The term "subject" as used in this application includes mammals and humans, preferably humans.

The term "effective amount" used in this application refers to an amount sufficient to obtain or at least partially obtain the desired effect. For example, a therapeutically effective amount refers to an amount sufficient to cure or at least partially prevent the disease and its complications in patients who have already suffered from the disease. It is completely within the abilities of those skilled in the art to determine such an effective amount. For example, the effective amount for therapeutic use will depend on the severity of the disease to be treated, the overall state of the patient’s own immune system, the patient’s general conditions such as age, weight and sex, the way the drug is administered, and other treatments that are administered simultaneously and many more.

Proxanthin, its stereoisomer, its solvate, its pharmaceutically acceptable salt, the hydrate or solvate of its pharmaceutically acceptable salt, or its pharmaceutically acceptable The amount of various modifications or second anti-tumor drugs received depends on the type and severity of the disease or condition and the characteristics of the subject, such as general health, age, sex, weight, and tolerance to the drug It also depends on factors such as the type of preparation and the way the drug is administered, as well as the administration cycle or time interval. Those skilled in the art can determine the appropriate dosage based on these and other factors. Generally speaking, proxanthin, its stereoisomer, its solvate, its pharmaceutically acceptable salt, the hydrate or solvate of its pharmaceutically acceptable salt, or its pharmaceutically acceptable The daily dosage of various modifications or second anti-tumor drugs for treatment can be about 0.0001-1000 mg/kg body weight/day, and the daily dosage can be administered once or in multiples as appropriate.

The beneficial technical effects of this application

The antitumor drug proxanthin provided in this application, such as proxanthin sulfate, has broad-spectrum therapeutic activity on lung cancer. The advantage of proxanthin is that it is easy to accept by patients, and it is cheap and easy to obtain. It is foreseeable that proxanthin will become a clinical drug that can be taken for a long time and can effectively inhibit the metastasis, invasion and recurrence of lung cancer.

Description of the drawings

Figure 1 shows the inhibitory effect of proxanthin sulfate on the proliferation of human lung adenocarcinoma cell lines;

Figure 2 shows the proliferation inhibitory effect of proxanthin sulfate on human lung squamous cell line;

Figure 3 shows the inhibitory effect of proxanthin sulfate on the proliferation of human large cell lung cancer cell lines;

Figure 4 shows the inhibitory effect of proxanthin sulfate on the proliferation of human small cell lung cancer cell lines;

Figure 5 shows that proxanthin sulfate can inhibit lung cancer cell migration;

Figure 6 shows that proxanthin sulfate can inhibit the growth of lung cancer cell lines in the SCID mouse model.

detailed description

The technical solutions in the embodiments of the present application will be clearly and completely described below in conjunction with the drawings in the embodiments of the present application. Obviously, the described embodiments are only a part of the embodiments of the present application, rather than all of the embodiments. Based on the embodiments in this application, all other embodiments obtained by a person of ordinary skill in the art fall within the protection scope of this application.

In the following examples, the test materials used and their sources include:

Protoxanthin sulfate, purchased from MCE; high glucose DMEM medium, RIPM1640 medium, fetal bovine serum FBS (Gibco), CCK-8 kit purchased from Biyuntian Biotechnology Co., Ltd.; Annexin V-FITC cell apoptosis detection reagent The box was purchased from Biyuntian Biotechnology Co., Ltd.; the cell cycle detection kit was purchased from Biyuntian Biotechnology Co., Ltd.; 96-well plates (Nunc brand) were purchased from Thermo Company; SCID mice were purchased from Shanghai Slack Laboratory Animal Co., Ltd.

In the following Examples 1-2, proxanthin sulfate was diluted with a culture medium to a mother liquor with a concentration of 10 mmol/L, and then diluted with a culture medium to use concentrations of different gradients during use. The proxanthin sulfate injection used in Example 3 was prepared using phosphate buffered saline (PBS) with a concentration of 200 μg/mL.

The cell line selected in the following examples is: human lung adenocarcinoma cell line A549 (medium: DMEM high glucose medium plus 10% FBS, purchased from

CCL-185); human lung adenocarcinoma cell H1299 (medium: DMEM high glucose medium plus 10% FBS, purchased from

CRL-5803); human lung adenocarcinoma cell H358 (medium: DMEM high glucose medium plus 10% FBS, purchased from

CRL-5807); human lung squamous cell carcinoma SK-MES-1 (medium: DMEM high glucose medium plus 10% FBS, purchased from

HTB-58); human lung squamous cell carcinoma cell Ebc-1 (medium: RIPM1640 plus 10% FBS, purchased from Thermo fisher scientific: 11875101); human lung squamous cell carcinoma cell H520 (medium: DMEM high glucose medium plus 10% FBS, purchased from

HTB-182); human lung squamous cell carcinoma cell H2170 (medium: DMEM high glucose medium plus 10% FBS, purchased from

CRL-5928); human lung squamous cell carcinoma cell H1703 (medium: DMEM high glucose medium plus 10% FBS, purchased from

CRL-5889); human large cell lung cancer cell H661 (medium: DMEM high glucose medium plus 10% FBS, purchased from

HTB-183); human small cell lung cancer cell H510A (medium: DMEM high glucose medium plus 10% FBS, purchased from

HTB-184); human small cell lung cancer cell H1417 (medium: DMEM high glucose medium plus 10% FBS, purchased from

CRL-5869); human small cell lung cancer cell DMS 114 (medium: DMEM high glucose medium plus 10% FBS, purchased from

CRL-2066).

Example 1 Experiment on the killing effect of proxanthin sulfate on different tumor cells of lung cancer

The experimental method is to determine the viability of CCK-8 cells. The CCK-8 reagent contains WST-8 [chemical name: 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfonic acid) Acid benzene)-2H-tetrazole monosodium salt], it is activated by the dehydrogenase in the cell under the action of the electron carrier 1-methoxy-5-methylphenazinium dimethyl sulfate (1-Methoxy PMS) It is reduced to a highly water-soluble yellow formazan product (Formazan dye). The amount of formazan produced is proportional to the number of living cells. Therefore, this feature can be used to directly analyze cell proliferation and toxicity.

The specific experimental procedure is: after the cells are digested with trypsin, they are dispersed into individual cells and suspended in the corresponding DMEM or RIPM1640 medium. The cells were seeded in a 96-well culture plate, 5000 cells/well. Place the cells in a 37°C/carbon dioxide (5%) incubator for 8 hours to allow the cells to adhere to the wall. On the second day, the culture solution was discarded, the cell culture solution of proxanthin sulfate was added (the specific concentration is mentioned in the results of each cell), and a control well without drug added was set. Place in a 37°C/carbon dioxide (5%) incubator for 48 hours, aspirate the supernatant, add 100μL of serum-free DMEM medium and 10μL of CCK-8 detection reagent to each well, 37°C/carbon dioxide (5%) Incubate for 2-4h. Measure the absorbance at 450nm/620nm with a microplate reader (Multiscan FC, product of Thermo), and calculate the cell survival rate by the following formula.

Cell survival rate = (experimental group OD/control group OD)*100%

Experimental results 1 Proxanthin sulfate inhibits proliferation of lung adenocarcinoma cell lines

The inhibitory effects of proxanthin sulfate on the proliferation of lung adenocarcinoma cell lines A549, H1299, and H358 are shown in Figure 1. The concentration of proxanthin sulfate used was 5 μmol/L. It can be seen from Figure 1 that the proliferation inhibitory effect of proxanthin sulfate on the three lung adenocarcinoma cells can reach more than 90%, confirming that proxanthin sulfate has a good proliferation inhibitory effect on lung adenocarcinoma cells.

Experimental results 2 Proliferation inhibitory effect of proxanthin sulfate on lung squamous cell carcinoma cell lines

The proliferation inhibitory effects of proxanthin sulfate on the lung squamous cell lines SK-MES-1, Ebc-1, H520, H2170, and H1703 are shown in Figure 2. The concentration of proxanthin sulfate used was 5 μmol/L. It can be seen from Figure 2 that the proliferation inhibitory effect of proxanthin sulfate on the five lung squamous cell carcinoma cells can reach more than 90%, confirming that proxanthin sulfate has a good proliferation inhibitory effect on lung squamous cell carcinoma cells.

Experimental Result 3 The inhibitory effect of proxanthin sulfate on the proliferation of large cell lung cancer cell lines

The inhibitory effect of proxanthin sulfate on the proliferation of large cell lung cancer cell line H661 is shown in Figure 3. The concentration of proxanthin sulfate used was 5 μmol/L. It can be seen from Figure 3 that the proliferation inhibitory effect of proxanthin sulfate on large cell lung cancer cells can reach more than 90%, confirming that proxanthin sulfate has a good proliferation inhibitory effect on large cell lung cancer cells.

Experimental results 4 Proxanthin sulfate inhibits the proliferation of small cell lung cancer cell lines

The proliferation inhibitory effect of proxanthin sulfate on small cell lung cancer cell lines H510A, H1417, DMS 114 is shown in Figure 4. The concentration of proxanthin sulfate used was 5 μmol/L. It can be seen from Fig. 4 that the inhibitory effect of proxanthin sulfate on the proliferation of small cell lung cancer cells can reach more than 90%, confirming that proxanthin sulfate has a good proliferation inhibitory effect on small cell lung cancer cells.

Example 2 Experiment of Protoxanthin Sulfate Inhibiting Tumor Cell Migration

The experimental method is a cell scratch test. The cell scratch test is the simplest and most economical in vitro test method for studying cell migration. The principle of this method is that when the cells grow to a monolayer state, a blank area is artificially created on the fused monolayer cells, called "scratches." The cells on the edge of the scratch will gradually enter the blank area to make the "scratch" heal. The stronger the cell migration ability, the shorter the time to heal the scratch.

The specific experimental procedure is as follows: digest the cells with trypsin for 3 minutes, and add culture medium to terminate the digestion reaction. Then mix the cells by pipetting gently, and evenly seed the cells in a six-well plate. The cells were incubated in a 37°C incubator containing 5% CO ₂ for 8-24 hours. The number of cells varies from cell to cell, and it should be fully covered overnight. On the next day, use the sterile tip of a 1mL pipette to compare with the straightedge, try to be perpendicular to the straightedge to make the scratch, and the tip should be vertical and not tilted. The most important thing for cell scratches is to draw the line as straight as possible, and the width of the cells in the experimental group and the control group are similar to ensure that the experimental group and the control group are comparable. Wash the cells gently with PBS 3 times to remove the marked cells. The experimental group was added with different concentrations of proxanthin sulfate serum-free culture solution, and the control group was added with the same volume of serum-free culture solution. Put it into a 37°C/5% CO ₂ incubator and cultivate. Sampling and taking pictures at 0, 12, and 24 hours. The direction of the "scratch" when taking pictures is best horizontal or vertical in the field of view.

Experimental results 5 Proxanthin sulfate can significantly inhibit the migration of lung cancer cells

The lung cancer cell line A549 can significantly inhibit the migration ability of tumor cells after being treated with cell culture solutions of different concentrations of proxanthin sulfate. The results are shown in Figure 5. The drug concentration of proxanthin sulfate is 1μmol/L, 2.5μmol/L, 5μmol/L. It can be seen from Figure 5 that compared to the control group, when the concentration of proxanthin sulfate is as low as 1 μmol/L, proxanthin sulfate can still significantly inhibit tumor cell migration.

Example 3 Experiment of Proxanthin Sulfate Inhibiting Proliferation of Lung Cancer Cells in SCID Mouse Model

Experimental animals: 6-8 week old female C.B17SCID mice, which are from Shanghai Slack Experimental Animal Co., Ltd.; according to the protocol approved by the Experimental Animal Center of Xiamen University and the Ethics Committee, the mice were under SPF conditions Rearing.

Specific experimental procedure: The tumor cells used for subcutaneous tumor formation in SCID mice (lung adenocarcinoma cell line A549) were digested with 0.01% trypsin, and then resuspended into single cells in a cell culture medium containing 10% fetal bovine serum Suspension; count the cell density of the suspension, centrifuge at 1000g for 3min to pellet the cells, and then resuspend the cells with an appropriate volume of PBS to reach about 10 ⁶ -10 ⁷ cells/100μL PBS; follow 10 ⁶ -10 ⁷ cells/ 100μL PBS/point was used to inoculate tumor cells subcutaneously on the back of SCID mice with a syringe. After about 14-21 days, when tumor cells formed a tumor mass of about 100mm ^{3 under} the skin of SCID mice, the tumor-bearing SCID mice were randomly divided into sulfuric acid Proxanthin treatment group and negative control group, 4 mice in each group, mice in the proflavin sulfate treatment group were injected intraperitoneally with proflavin sulfate injection at a dose of 20 mg/kg, and the negative control group was injected with the same volume without The PBS solution of proxanthin sulfate was injected once every two days for a total of 5 injections. Use vernier calipers to measure and record tumor size changes every two days. The calculation method of tumor size is:

Tumor size (mm ³ ) = tumor length value x (tumor width value) ² /2.

Experimental results 6 Proxanthin sulfate treatment can effectively inhibit the proliferation of tumor cells in the SCID mouse model

The growth and changes of tumor volume in each mouse in the proxanthin sulfate treatment group and the negative control group are shown in Figure 6 respectively. The results showed that the tumor growth rate of the SCID mice in the proxanthin sulfate treatment group was slower than that in the SCID mice in the control group, and the tumor size in the treatment group was much smaller than the control group. The above results indicate that proxanthin sulfate has good lung cancer therapeutic activity.

Through the above experiments, we can see the broad spectrum and effectiveness of proxanthin sulfate as a therapeutic drug for lung cancer. At the same time, the proxanthin sulfate of the present application has incomparable advantages over other drugs, and it has good safety, low price, and easy access. In addition, proxanthin sulfate will become a clinical drug that can be taken for a long time and can effectively inhibit tumor metastasis, invasion and recurrence due to its good safety and effectiveness.

The above descriptions are only the preferred embodiments of this application, and are not intended to limit this application. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of this application shall be included in this application. Within the scope of protection.

Claims

Proxanthin, its stereoisomers, its solvates, its pharmaceutically acceptable salts, the hydrates or solvates of its pharmaceutically acceptable salts, or its various pharmaceutically acceptable modifications Use in the preparation of medicines for treating lung cancer.
Use of a pharmaceutical composition in the preparation of a medicament for the treatment of lung cancer, wherein the pharmaceutical composition contains proxanthin, its stereoisomers, its solvates, its pharmaceutically acceptable salts, and its pharmaceutically acceptable The hydrate or solvate of the salt, or various pharmaceutically acceptable modifications thereof, and a pharmaceutically acceptable carrier or excipient.
Proxanthin, its stereoisomers, its solvates, its pharmaceutically acceptable salts, the hydrates or solvates of its pharmaceutically acceptable salts, or its various pharmaceutically acceptable modifications , Which is used to treat lung cancer.
A pharmaceutical composition for the treatment of lung cancer, wherein the pharmaceutical composition contains proxanthin, its stereoisomers, its solvates, its pharmaceutically acceptable salts, and the hydrates of its pharmaceutically acceptable salts Or solvates, or various pharmaceutically acceptable modifications thereof, and pharmaceutically acceptable carriers or excipients.
Contains proxanthin, its stereoisomers, its solvates, its pharmaceutically acceptable salts, the hydrates or solvates of its pharmaceutically acceptable salts, or various pharmaceutically acceptable modifications thereof A combination product of a drug and a second antitumor drug, which is used to treat lung cancer.
A method for treating lung cancer, comprising: administering a therapeutically effective amount of proxanthin, its stereoisomers, its solvates, its pharmaceutically acceptable salts, and its pharmaceutically acceptable amounts to a subject in need Accepted salt hydrates or solvates, or various pharmaceutically acceptable modifications thereof.
A method for treating lung cancer, comprising: administering a therapeutically effective amount of a pharmaceutical composition to a subject in need thereof, wherein the pharmaceutical composition contains proxanthin, its stereoisomers, its solvates, and A pharmaceutically acceptable salt, a hydrate or solvate of a pharmaceutically acceptable salt thereof, or various pharmaceutically acceptable modifications thereof, and a pharmaceutically acceptable carrier or excipient.
A method for treating lung cancer, comprising: administering a therapeutically effective amount of proxanthin, its stereoisomers, its solvates, its pharmaceutically acceptable salts, and its pharmaceutically acceptable amounts to a subject in need Accepted salt hydrates or solvates, or various pharmaceutically acceptable modifications thereof, and a second anti-tumor drug with therapeutic sales.
The use of claim 2, the pharmaceutical composition of claim 4, or the method of claim 7, wherein the pharmaceutical composition further contains a second anti-tumor drug.
The use of claim 2, the pharmaceutical composition of claim 4, or the method of claim 7, wherein the pharmaceutical composition is a tablet, capsule, pill, oral liquid preparation, granule, powder or injection.
The use of claim 2, the pharmaceutical composition of claim 4, or the method of claim 7, wherein the pharmaceutical composition is administered by oral, injection, implant, topical, spray or inhalation.
Proxanthin, its stereoisomers, its solvates, its pharmaceutically acceptable salts, the hydrates or solvates of its pharmaceutically acceptable salts, or its various pharmaceutically acceptable modifications It is used in combination with a second anti-tumor drug to prepare a drug for treating lung cancer.
The use of any one of claims 1 to 12, proxanthin, its stereoisomers, its solvates, its pharmaceutically acceptable salts, the hydrates or solvates of its pharmaceutically acceptable salts, or Various pharmaceutically acceptable modifications, pharmaceutical compositions, combined products or methods thereof, wherein:

For example, the pharmaceutically acceptable salt is proxanthin sulfate or proxanthin hydrochloride,

For example, the hydrate of the pharmaceutically acceptable salt is proxanthin sulfate hydrate or proxanthin hydrochloride hydrate.
The use of claim 3, 9 or 12, the pharmaceutical composition of claim 9, the combination product of claim 5, the method of claim 8 or 9, wherein the second anti-tumor drug is an immune checkpoint inhibitor such as PD -1 inhibitors (including Nivolumab, Pembrolizumab, IBI-308, Camrelizumab (SHR-1210), Tislelizumab, Cemiplimab, BCD-100, TSR-042, PDR-001, JNJ-63723283), PD-L1 inhibitors (including Atezolizumab , Avelumab, Durvalumab, BMS-936559, M-7824, CX-072), CTLA-4 inhibitors (including Ipilimumab, Tremelimumab, AGEN-1884, AK-104), etc. Traditional chemotherapy drugs such as Paclitaxel, cisplatin, 5-fluorouracil, etc., or small molecule inhibitors such as EGFR inhibitors (including Cetuximab, Erlotinib, Gefitinib, Lapatinib Ditosylate, Neratinib, Theliatinib, Cyasterone, Osimertinib, Avitinib, Afatinib, Gefitinib, Canertinib, Lapatinib 490, Pelitinib, Dacomitinib), HER2 inhibitors (including Poziotinib, Trastuzumab, Pertuzumab, Irbinitinib), ALK inhibitors (including Crizotinib, TAE684 (NVP-TAE684), Alectinib, AZD3463, ASP3026, Brigatinib (AP26113), XEnsartinib 396), Alectinib), etc.
The use of any one of claims 1 to 14, proxanthin, its stereoisomers, its solvates, its pharmaceutically acceptable salts, the hydrates or solvates of its pharmaceutically acceptable salts, or Various pharmaceutically acceptable modifications, pharmaceutical compositions, combined products or methods thereof, wherein the lung cancer is selected from: lung squamous cell carcinoma, lung adenocarcinoma, large cell lung cancer, small cell lung cancer or any combination thereof.
The use of any one of claims 1 to 14, proxanthin, its stereoisomers, its solvates, its pharmaceutically acceptable salts, the hydrates or solvates of its pharmaceutically acceptable salts, or Various pharmaceutically acceptable modifications, pharmaceutical compositions, joint products or methods thereof, wherein proxanthin, its stereoisomers, its solvates, its pharmaceutically acceptable salts, and its pharmaceutically acceptable The hydrate or solvate of the salt of, or various pharmaceutically acceptable modifications thereof are used as cytotoxic drugs, tumor growth inhibitors, anti-tumor metastasis drugs and/or anti-tumor invasion drugs.