CN113350325B - Application of dianthrone compound in preparation of antitumor drugs - Google Patents

Abstract

The invention provides an application of dianthrone compounds in preparing anti-liver cancer tumor drugs, belonging to the technical field of biological medicines; the dianthranone compound comprises Trans-emodin dianthranone and/or Cis-emodin dianthranone. The invention provides a new application of dianthrone compounds in the aspect of preventing and/or treating liver cancer. Experiments prove that the dianthrone compound has the following significant activity in pharmacology: inhibiting the growth of liver cancer cells, inducing S-phase arrest, inducing the apoptosis of liver cancer cells, and showing significant anti-liver cancer activity in vivo.

Description

Application of dianthrone compounds in the preparation of antitumor drugs

technical field

The invention relates to the technical field of biomedicine, in particular to the application of dianthrone compounds in the preparation of antitumor drugs.

Background technique

Liver cancer is a malignant tumor of the liver, which can be divided into two categories: primary and secondary. Liver cancer is an important public health problem in the world. The treatment of liver cancer mainly includes liver transplantation, liver cancer surgery, radiotherapy and drug chemotherapy. Surgery is the main treatment for liver cancer. However, due to the insidious onset of liver cancer, most patients are diagnosed at an advanced stage and are prone to liver cirrhosis. Less than 30% of patients can be treated with surgery, and the surgical resection rate is very low. The recurrence rate is high and the prognosis is extremely poor. Liver transplantation is also an effective treatment, but the indications for liver transplantation are extremely strict, and suitable liver sources are difficult to obtain. Among the other two treatments, liver cancer is less sensitive to radiotherapy and less sensitive to drug therapy, but the 5-year survival rate of inoperable liver cancer patients is only 5%, and drug therapy is still indispensable for liver cancer.

Cytotoxic drugs in conventional drug therapy, such as fluorouracil, doxorubicin, cisplatin, etc., not only have serious side effects in the treatment of liver cancer, but usually cannot significantly alleviate the progression of liver cancer or prolong life. At present, the only drug that has been proven to prolong the survival of patients with liver cancer is the tyrosine kinase inhibitor sorafe, which has significantly reduced toxic and side effects, and has poor efficacy and drug resistance.

At present, drugs for the treatment of liver cancer are far from meeting the clinical needs, and it is of great significance to discover new drugs with anti-liver cancer activity.

Contents of the invention

The purpose of the present invention is to provide the application of the dianthrone compounds in the preparation of anti-tumor drugs, and provide the new application of the dianthrone compounds in the prevention and/or treatment of liver cancer.

In order to achieve the above-mentioned purpose of the invention, the present invention provides the following technical solutions:

The invention provides the application of dianthrone compounds in the preparation of anti-liver cancer drugs;

The dianthrone compounds include Trans-emodin dianthrone and/or Cis-emodin dianthrone.

Preferably, when the dianthrone compounds include Trans-emodin dianthrone and Cis-emodin dianthrone, the molar ratio of Trans-emodin dianthrone and Cis-emodin dianthrone is 1: (0.5~1).

Preferably, the Trans-emodin dianthrone and Cis-emodin dianthrone include racemates of Trans-emodin dianthrone and Cis-emodin dianthrone.

Preferably, the actual racemization ratio of the racemate of Trans-emodin dianthrone and Cis-emodin dianthrone is 56:38.

Preferably, the liver cancer tumor is formed by human liver cancer HepG2 cells or mouse H22 liver cancer.

Preferably, the mass percent content of dianthrone compounds in the drug is 0.1%-95%.

Preferably, the mass percent content of dianthrone compounds in the drug is 30%-90%.

Preferably, the dosage form of the drug includes oral preparations, external preparations, injections or cavity administration preparations.

The invention provides the application of dianthrone compounds in the preparation of anti-liver cancer drugs; the dianthrone compounds include Cis-emodin dianthrone and/or Trans-emodin dianthrone. The invention provides a new application of the above-mentioned dianthrone compounds in the preparation of drugs for preventing and/or treating liver cancer. Experiments have proved that dianthrone compounds have the following significant pharmacological activities: inhibiting the growth of liver cancer cells, inducing S-phase arrest, inducing apoptosis of liver cancer cells, and exhibiting significant anti-liver cancer activity in vivo.

Description of drawings

Fig. 1 is to the growth inhibitory activity of human liver cancer HepG2 cell (n=4);

The influence of Fig. 2 on human liver cancer HepG2 cell cycle;

Figure 3 is the statistical result of the number of cells in S phase of human liver cancer HepG2 cells;

Fig. 4 The effect of promoting apoptosis of human liver cancer HepG2 cells.

Detailed ways

The invention provides the application of dianthrone compounds in the preparation of anti-liver cancer drugs;

The dianthrone compounds include Trans-emodin dianthrone and/or Cis-emodin dianthrone.

In the present invention, when the dianthrone compounds include Trans-emodin dianthrone and Cis-emodin dianthrone, the moles of the Trans-emodin dianthrone and Cis-emodin dianthrone The ratio is preferably 1:(0.5-1).

In the present invention, the chemical structural formula of the Trans-emodin dianthrone (HSW-W-25) is shown in formula I; the chemical structural formula of the Cis-emodin dianthrone (HSW-W-26) is as follows Shown in formula II;

In the present invention, there is no special limitation on the sources of Cis-emodin dianthrone and Trans-emodin dianthrone, which can be obtained from conventional commercial products or prepared by methods known in the art.

In the present invention, the Trans-emodin dianthrone and Cis-emodin dianthrone are preferably racemates comprising Trans-emodin dianthrone and Cis-emodin dianthrone, and the actual racemic ratio is preferably It is 56:38.

In the present invention, the liver cancer tumor is preferably formed by HepG2 cells or H22 cells.

In the present invention, the mass percentage of the dianthrone compound in the drug is preferably 0.1%-95%, more preferably 30%-90%, and most preferably 50%-80%.

In the present invention, the route of administration of the drug includes enteral administration or parenteral administration; the enteral administration includes oral administration or rectal administration; the parenteral administration includes: intravenous injection , intramuscular injection, subcutaneous injection, nasal cavity, oral mucosa, eyes, lungs and respiratory tract, skin or vagina.

In the present invention, the dosage form of the drug preferably includes oral preparations, external preparations, injections or cavity administration preparations.

In the present invention, the oral preparations preferably include tablets, capsules, pills, granules, oral liquids or suspensions; in the present invention, the tablets are preferably included in the present invention, so The injections preferably include aqueous injections, powder injections and transfusions.

In the present invention, the dosage form of the drug includes a gas dosage form, a liquid dosage form, a solid dosage form or a semi-solid dosage form.

In the present invention, the gas dosage forms preferably include aerosol or spray.

In the present invention, the liquid dosage forms include solutions, emulsions, suspensions, injections, eye drops, nasal drops, lotions or liniments; the solutions include true solutions or colloidal solutions; the emulsions include o/w type, w/o type revitalizing milk; the injection includes water injection, powder injection or infusion.

In the present invention, the solid dosage form preferably includes tablet, capsule, granule, powder, pellet, drop pill, suppository, film, patch or powder spray; in the present invention, the tablet Preferred include ordinary tablets, enteric-coated tablets, buccal tablets, dispersible tablets, chewable tablets, effervescent tablets or orally disintegrating tablets; in the present invention, the capsules preferably include hard capsules, soft capsules or enteric-coated capsules.

In the present invention, the semisolid dosage form preferably includes ointment, gel or paste.

In the present invention, the dosage form of the drug preferably includes common preparations, sustained-release preparations, controlled-release preparations, targeted preparations or microparticle drug delivery systems.

In the present invention, the drug preferably also includes pharmaceutically acceptable auxiliary materials. In the present invention, when the dosage form of the drug is a tablet, the auxiliary materials preferably include one or more of diluents, binders, wetting agents, disintegrants, lubricants and glidants.

In the present invention, the diluent preferably includes one of starch, dextrin, sucrose, glucose, lactose, mannitol, sorbitol, xylitol, microcrystalline cellulose, calcium sulfate, calcium hydrogen phosphate and calcium carbonate one or more; the wetting agent preferably includes one or more of water, ethanol and isopropanol; the binder preferably includes starch slurry, dextrin, syrup, honey, glucose solution, microcrystalline Cellulose, Arabic mucilage, gelatin pulp, sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, ethylcellulose, acrylic resins, carbomer, polyvinylpyrrolidone, and polyethylene glycol One or more of them; the disintegrant preferably includes dry starch, microcrystalline cellulose, low-substituted hydroxypropyl cellulose, cross-linked polyvinylpyrrolidone, cross-linked sodium carboxymethyl cellulose, carboxymethyl One or more of sodium starch, sodium bicarbonate, citric acid, polyoxyethylene sorbitol fatty acid ester and sodium dodecylsulfonate; the lubricant and glidant independently preferably include talcum powder One or more of , silicon dioxide, stearate, tartaric acid, liquid paraffin and polyethylene glycol.

In the present invention, the tablet is preferably made into a coated tablet, and the coated tablet preferably includes a sugar-coated tablet, a film-coated tablet or an enteric-coated tablet; in the present invention, the tablet Preferable ones include single-layer sheets or multi-layer sheets.

In the present invention, when the dosage form of the medicine is a capsule, the auxiliary materials preferably include one or more of diluents, glidants, wetting agents, binders and disintegrants.

In the present invention, when the dosage form of the drug is an injection, one or more of the water, ethanol, isopropanol and propylene glycol of the injection; the auxiliary material preferably includes a solubilizer, a cosolvent, a pH regulator and one or more of the osmotic pressure regulator; the solubilizer and co-solvent independently preferably include one or more of poloxamer, lecithin and hydroxypropyl-β-cyclodextrin; the The pH regulator preferably includes one or more of phosphate, acetate, hydrochloric acid and sodium hydroxide; the osmotic pressure regulator preferably includes sodium chloride, mannitol, glucose, phosphate and acetic acid One or more types of salt.

In the present invention, when the dosage form of the drug is preferably freeze-dried powder injection, the auxiliary material preferably includes a proppant; the proppant preferably includes mannitol and/or glucose.

In the present invention, the auxiliary materials of the drug preferably also include one or more of colorants, preservatives, spices and flavoring agents.

The present invention has no special limitation on the administration method of the drug, and any administration method known in the art can be adopted.

In the present invention, the daily dosage of the drug is preferably 0.001-150 mg/kg body weight, more preferably 0.1-100 mg/kg body weight, further preferably 1-60 mg/kg body weight, most preferably 2-30 mg/kg weight. The above-mentioned dosage can be administered in one dosage unit or divided into several dosage units, depending on the clinical experience of the doctor and the dosage regimen including the use of other therapeutic means.

The medicine of the present invention can be taken alone, or used in combination with other therapeutic medicines or symptomatic medicines. When the drug of the present invention has a synergistic effect with other therapeutic drugs, its dosage should be adjusted according to the actual situation.

The technical solutions in the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Apparently, the described embodiments are only some of the embodiments of the present invention, but not all of them. Based on the embodiments of the present invention, all other embodiments obtained by persons of ordinary skill in the art without making creative efforts belong to the protection scope of the present invention.

The following examples and pharmacological activity experiments are used to further illustrate the present invention, but this does not imply any limitation to the present invention. For the experimental methods that do not specify specific conditions in the following examples, select according to conventional methods and conditions, or according to the product instructions.

In the following examples, the full names or corresponding Chinese names of some substances are as follows:

DHS: Emodin

MTT: thiazole blue

DMEM: a medium containing various amino acids and glucose

DMSO: dimethyl sulfoxide

PI: Propidium Iodide, propidium iodide

AnnexinV/FITC: Apoptosis Detection Reagent

The room temperature described in the following examples is as conventionally described in the art, and generally refers to 15-25°C.

Experimental example 1 Test of growth inhibitory activity of HSW-W-25 and HSW-W-26 on human liver cancer HepG2 cells

experimental method:

Take the human liver cancer HepG2 cells in the logarithmic growth phase, inoculate ⁵ ×104 cells/ml in a 96-well plate, 100 μl per well, culture for 24 hours, add HSW-W-25 (10 μM, 20 μM), HSW-W -26 (10 μM, 20 μM), positive control drug DHS (25 μg/ml), act on the cells for 48 hours. After removing the supernatant, add 100 μl MTT (0.5 mg/ml, prepared using serum-free DMEM culture solution) to each well, put it in a cell culture incubator and cultivate it for 4 hours, add 150 μl DMSO to each well, and measure the absorbance at 570 nm in a microplate reader. Calculate cell viability.

Cell survival rate (%)=100*(OD value of the blank control group-OD value of the administration group)/OD value of the blank control group.

The experimental results are as follows:

HSW-W-25 (10 μM, 20 μM) and HSW-W-26 (10 μM, 20 μM) acted on human liver cancer HepG2 cells for 48 hours, and showed significant inhibitory activity on the growth of liver cancer cells. See Figure 1.

Experimental example 2 HSW-W-25 and HSW-W-26 induce human liver cancer HepG2 cell cycle arrest test

experimental method:

Take the human liver cancer HepG2 cells in the logarithmic growth phase, inoculate 1.5×10 ⁵ cells/ml in a six-well plate, 1 ml per well, and after culturing for 24 hours, add HSW-W-25 (10 μM, 20 μM), HSW-W -26 (10 μM, 20 μM), DHS (25 μg/ml), continue to act on the cells for 48 hours. Centrifuge at 1000rpm for 3min to collect the cells, wash the cells once with 4°C pre-cooled PBS, add 1mL DNA staining solution and 10μl permeabilization solution, vortex for 5-10 seconds to mix, incubate at room temperature for 30min in the dark, flow cytometry (FACS ) for cell cycle analysis.

The experimental results showed that when HSW-W-25 (10 μM, 20 μM) and HSW-W-26 (10 μM, 20 μM) were applied to human liver cancer HepG2 cells for 48 hours, the cell cycle dynamics changed significantly, and the proportion of cells in S phase increased significantly (Figure 2 , Figure 3), indicating that HSW-W-25 and HSW-W-26 inhibit the proliferation of liver cancer cells and induce S-phase arrest.

Experimental example 3 HSW-W-25 and HSW-W-26 induce apoptosis of human liver cancer HepG2 cells

experimental method:

Take human liver cancer HepG2 cells in the logarithmic growth phase, inoculate 1.5×10 ⁵ cells/ml in a six-well plate, 1 ml per well, and after culturing for 24 hours, add HSW-W-25 (10 μM, 20 μM), HSW-W -26 (10 μM, 20 μM), DHS (25 μg/ml) for 48 hours, centrifuge at 1000 rpm for 3 minutes, collect the cells, wash the cells twice with 4°C pre-cooled PBS, resuspend the cells in 500 μl binding buffer, and then add 5 μl AnnexinV/FITC solution and 10 μl PI solution, mixed well, and incubated at room temperature in the dark for 5 minutes, and analyzed cell apoptosis by flow cytometry (FACS).

The experimental results showed that HSW-W-25 (10 μM, 20 μM) and HSW-W-26 (10 μM, 20 μM) treated human liver cancer HepG2 cells for 48 hours, could significantly induce apoptosis of liver cancer cells, and the ratio of apoptotic cells was significantly higher than that of the blank control group. High (Figure 4).

Experimental example 4 Test of tumor inhibitory activity of HSW-W-25 and HSW-W-26 on mice bearing H22 liver cancer

experimental method:

Kunming mice were inoculated with 0.2ml H22 liver tumor liquid in the anterior armpit, and randomly divided into groups the next day, positive control drug 5-FU 40mg/kg group, HSW-25/26 20mg/kg group and blank control group, 10 mice in each group. HSW-25/26 was administered by intragastric administration for ten consecutive days, and 5-FU was administered by intraperitoneal injection once every three days, a total of three times. The mice were sacrificed 24 hours after the last administration, the final body weight and tumor weight were recorded, and the tumor inhibition rate was calculated. Tumor inhibition rate (%)=100*(tumor weight of blank control group-tumor weight of administration group)/tumor weight of blank control group.

The experimental results are shown in Table 1:

HSW-W-25, the racemic form of HSW-W-26 HSW-W-25/26, 20 mg/kg for 10 consecutive days, has significant antitumor activity against H22 liver cancer, and the tumor weight is significantly lower than that of the blank control group , while the body weight of the animals did not decrease significantly, indicating that the animals tolerated HSW-W-25/26 well (Table 1).

Table 1. Tumor inhibitory activity to mice bearing H22 liver cancer (n=10)

The above is only a preferred embodiment of the present invention, it should be pointed out that, for those of ordinary skill in the art, without departing from the principle of the present invention, some improvements and modifications can also be made, and these improvements and modifications can also be made. It should be regarded as the protection scope of the present invention.

Claims (6)

1. The application of dianthrone compounds in preparing anti-liver cancer and anti-tumor drugs;

the dianthranone compounds are Trans-emodin dianthranone and Cis-emodin dianthranone; the molar ratio of the Trans-emodin dianthrone to the Cis-emodin dianthrone is 1:1; the Trans-emodin dianthrone and the Cis-emodin dianthrone are racemates of the Trans-emodin dianthrone and the Cis-emodin dianthrone.

2. The use according to claim 1, wherein the actual racemization ratio of racemates of Trans-emodin dianthrone and Cis-emodin dianthrone is 56:38.

3. the use of claim 1 or 2, wherein the liver cancer tumor is formed by human liver cancer HepG2 cells or mouse H22 liver cancer.

4. The use according to claim 1 or 2, wherein the content of dianthrone compounds in the medicament is 0.1-95% by mass.

5. The use according to claim 4, wherein the content of dianthrone compounds in the medicament is 30-90% by mass.

6. The use according to any one of claims 1, 2 and 5, wherein the medicament is in a dosage form selected from the group consisting of oral, topical, injectable and transluminal.

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