WO2015090139A1

WO2015090139A1 - Antitumor drug, preparation method and use thereof

Info

Publication number: WO2015090139A1
Application number: PCT/CN2014/092627
Authority: WO
Inventors: 李徐钦
Original assignee: 李徐钦; 马丹霞
Priority date: 2013-12-20
Filing date: 2014-12-01
Publication date: 2015-06-25
Also published as: CN103655822B; CN103655822A

Abstract

An antitumor drug, consisting of ferula asafoetida, toad, horse bezoar and spreading hedyotis herb by weight ratio of 30:30:50:150. The method of preparing the drug comprises the following steps: weighing the raw materials, pulverizing, screening and blending. The drug can be used for treating cancer, such as liver cancer, stomach cancer or esophagus cancer. The drug has good anti-cancer effect, and can inhibit approximately 90% of tumor cells.

Description

Antitumor drug, preparation method and use thereof

Technical field

The invention relates to the field of traditional Chinese medicine, in particular to an antitumor medicine, a preparation method and the use thereof.

Background technique

Cancer is one of the major diseases that seriously threaten human health and social development. It is also a major disease that threatens the health of our residents. Recent statistics show that the incidence of cancer in China is increasing year by year. Among them, cancers such as gastric cancer and colon cancer account for more than 10% of all cancers, which are common tumors with greater harm. At present, the commonly used radiotherapy and chemotherapy methods in the treatment of tumors, while killing tumor cells, also cause the body's bone marrow suppression and immune function is low, and the side effects are large. Therefore, searching for high-efficiency and low-toxic anti-tumor drugs or active ingredients from natural animals, plants and minerals is a hot topic in recent years. Traditional Chinese medicine is a unique treasure house of traditional medicine in China. It has been used in disease prevention and treatment in China for more than 2,000 years and provides a very rich resource for the development of anticancer drugs.

At present, the highest cancer cell inhibition rate detected by similar drugs in China is only about 50%, and the cancer suppression rate of the well-known valuable anti-cancer drug Taxus is only about 30%.

Therefore, there is an urgent need to develop more effective anti-tumor drugs.

Summary of the invention

It is an object of the present invention to provide a more effective anti-tumor Chinese medicine.

In order to achieve the above object, the present invention provides an antitumor drug characterized by being composed of the following components in a weight ratio of Awei: 蟾蜍: Mabao: Hedyotis diffusa = 30:30:50:150.

The method for preparing the antitumor drug is characterized in that each component is weighed, and each component is pulverized, sieved, and mixed. The components of the anti-tumor drug obtained by the screening have a diameter of 300-800 mesh. The specific steps can be carried out in accordance with a conventional method.

The invention also protects the use of the anti-tumor drug for anti-tumor. The method includes feeding the antitumor drug with water or rice wine.

The tumor is liver cancer, gastric cancer, and esophageal cancer.

The anti-tumor drug of the present invention, that is, "Li's glutinous food powder" is based on the experience of ancestral medical practice and absorbs various therapeutic esophagus of various generations. The theoretical essence of cancer, the formation of a unique experience of the experience side. "Lishi 膈食散" is made up of Chinese medicines such as Awei, 蟾蜍, Mabao, and Hedyotis diffusa according to the theory of traditional Chinese medicine. It has been used clinically for decades and has been observed to have significant effects on cancers of the digestive tract such as esophageal cancer and gastric cancer. It can effectively eliminate cancer lumps, alleviate the suffering of patients, and promote appetite and prolong the life of cancer patients.

The "Li's glutinous food" compound is mainly composed of traditional Chinese medicines such as A., A., and Hedyotis diffusa. A Wei is a commonly used traditional Chinese medicine, as recorded in the "New Revision of the Materia Medica". Awei has a special odor, tastes bitter and pungent, has the effect of reducing qi, promoting blood circulation, sputum and excitatory nerves. Zhao Bei et al. preliminarily proved that the ferulic medicinal liquor has certain anti-tumor effects through the three levels of whole body, cell and molecule, mainly by preventing the infiltration and metastasis of the tumor and improving the immunity of the body. The dagger was contained in the "Medical Doctors", and the "Shen Nong's Herbal Classic" contained the sexual taste and indications of "Shrimp." Ancient and modern prescriptions are widely used in the treatment of phlegm, phlegm and phlegm, detoxification, swelling, pain, and diuretic. It has been used in the clinical anti-tumor, analgesic, diuretic swelling and other preparations such as Qishen Zhentong Ointment, Zhishen Injection, Huachansu Injection and Fufang Suede Capsule. Hedyotis diffusa is sweet, light, cool, and belongs to the stomach, large intestine, small intestine, has the effect of clearing away heat and detoxification, reducing swelling and analgesia, and attending appendicitis, bronchitis, urinary tract infection, snake bite, tumor, intestinal wind and blood. Wait. In vitro experiments showed that ethanol extract of Hedyotis diffusa on human poorly differentiated gastric adenocarcinoma cells (BGC), human oral epidermoid carcinoma cells (KB), human hepatoma cells (SMMC), human cervical cancer cells (HELA), human lung cancer cells ( A549) has a certain inhibitory effect. In vivo, ethanol extract of Hedyotis diffusa has a certain inhibitory effect on transplanted S180 solid tumor in mice. The clinical preparation of the liver preparation in the middle and late stage of treatment of advanced liver cancer with the clinical preparation of Hedyotis diffusa as the medicinal herb can significantly improve the quality of life and prolong the survival time of patients with stage III primary liver cancer. The main medicinal taste of "Li's glutinous food" has good anti-tumor effect both in clinical trials and experimental studies, indicating that the "Li's glutinous food" compound compatibility has the basis of theoretical and practical application.

"Li's glutinous food" compound has both animal and botanical drugs. The main active ingredients of each drug are different. The main active ingredient of cockroach anti-tumor is scorpion lactone, which is a cardiac glycoside, including Bufalin, resibufogenin, scorpion venom (also known as cinobufagin); the main active components of ferula are volatile oil, coumarin and phenylpropanoid, white flower snake tongue The main active ingredients of grass are flavonoids, triterpenoids, organic acids and polysaccharides. Mabao can detoxify and remove sores. These different active ingredients play a role in different tumor targets, fully reflecting the multi-component and multi-target of traditional Chinese medicine. The characteristics of the action, from the overall adjustment of the body function to achieve the purpose of removing disease.

The "Li's glutinous food" clinical trial uses a powder, which is to smash, sift and mix the medicinal materials, which is used for the treatment of patients with digestive tract cancer such as gastric cancer and esophageal cancer. The patient's taking method is usually served with rice wine. Since the medicinal material powder cannot enter the cell during in vitro cell experiments, the medicinal material must be extracted. The invention provides two methods for water extraction and alcohol extraction of "Li's glutinous food powder", and the anti-tumor activity of both extracts is studied in vitro. The results showed that the two extracts had anti-tumor effects on liver cancer cells (HpeG-2), gastric cancer cells (MKN-45) and esophageal cancer cells (EC-109), and the effects of ethanol extract were obtained in vitro. better. The maximum inhibition rate of ethanol extracts on three tumor cell lines was about 90%. The results of the present invention preliminarily indicate that the "Li's glutinous food" fat-soluble component may have better anti-tumor activity. It also indirectly proves that there is a scientific basis for the use of rice wine in clinical trials.

The anti-tumor drug of the invention has been subjected to an in vitro anti-tumor test research report, and the test data shows that the "Li's glutinous powder" water extract and the ethanol extract are for liver cancer cells (HpeG-2), gastric cancer cells (MKN-45), Three digestive system cell lines of esophageal cancer cells (EC-109) have obvious anti-tumor effects in vitro. The ethanol extract was more effective, with a maximum inhibition rate of nearly 90% for the three cell lines. The maximum inhibition rate of liver cancer cells was 88.40%, the maximum inhibition rate to esophageal cancer cells was 91.88%, and the maximum inhibition rate to gastric cancer cells was 93.98%. The highest cancer cell inhibition rate detected by similar drugs in China is only about 50%, and the cancer suppression rate of the well-known valuable anti-cancer drug Taxus is only about 30%. The maximum inhibition rate of the cancer cells of the present invention is much higher than this.

Because Chinese medicine is very particular about the accuracy of the dose of the drug, the theory of compatibility of Junchen Zuo, etc., when the dose of the drug is changed, it will have a corresponding impact on the effect of the entire prescription. It can be seen from the effect verification of Example 2 that when any component in the formulation changes, the effect is greatly affected, and the anticancer effect is greatly reduced. Therefore, it is further proved that the formulation of the present invention is obtained after years of experience and creative work of the applicant. Has a very good anti-cancer effect.

detailed description

The embodiments of the present invention are described in detail below, and the examples of the embodiments are intended to explain the present invention and are not to be construed as limiting. Where specific techniques or conditions are not indicated in the examples, they are carried out according to the techniques or conditions described in the literature in the art or in accordance with the product specifications. The reagents or instruments used are not indicated by the manufacturer, and are conventional products that can be obtained commercially.

Commercially available materials used in the examples:

Cells: human hepatoma cells (HepG-2), gastric cancer cells (MKN-45), and esophageal cancer cells (EC-109) were purchased from the Cell Resource Center of the Institute of Basic Medical Sciences of the Chinese Academy of Medical Sciences.

Reagents: 95% ethanol (analytical grade, Beijing Chemical Plant), DMSO (analytical grade, Beijing Chemical Plant), RPMI 1640 medium (Solarbio, China), trypsin (Solarbio, China), fetal bovine serum (Gibco) , USA), phosphate buffer (Thermo, Germany), MTT reagent (Gibco, USA);

Instruments: pure water meter (Millipore, USA), 96-well cell culture plate (Thermo, Germany), inverted microscope (OLYMPUS, Japan), carbon dioxide incubator (Thermo, Germany), continuous wavelength microplate reader (BIO) -TEK, USA).

Example 1: Preparation method of antitumor drug (Lee's glutinous food) of the present invention

Ingredients: Awei 30g 蟾蜍 30g Ma Bao 50g white flower Hedyotis 150g.

Step: Weigh the medicinal materials, crush and sieve to obtain 300-800 mesh powder, and mix well.

1) Preparation method of Lishi 膈 food and water extract

Li's granules were granulated with powder 10g, added with 100ml of water and refluxed three times for 30min each time, filtered, and the filtrate was combined and dried in vacuo to obtain a water extract of Lishi glutinosa, the yield was 16.8%.

2) Preparation method of alcohol extract of Lee's cockroach

10 g of Li's foraging powder was weighed and added with 100 ml of 95% ethanol solution for three times, each time for 30 min, filtered, and the filtrate was combined and dried in vacuo to obtain a Li's glutinous food ethanol extract with a yield of 10.5%.

Example 2: Antitumor effect experiment

Hepatoma cell line (HepG-2), gastric cancer cell line (MKN-45) and esophageal cancer cell line (EC-109) in logarithmic growth phase were digested and counted in 96-well culture plates at a rate of approximately 8,000 per well. -10000 cells, incubate at 37 ° C for 12 hours, discard the culture solution, add 100 μL of different concentrations of drug-containing medium (configured with RPMI 1640 medium), continue to incubate at 37 ° C for 24 hours, aspirate the culture solution, and add 150 μL per well. Phosphate buffer solution and 100 μL of 0.5g·L ^-1 MTT solution, continue to incubate at 37 ° C for 4 hours, then discard the MTT solution and phosphate buffer solution, add 150 μL of DMSO to each well, and shake the plate for 15 min. The absorbance value (OD) of each well was measured by the wavelength of 570 nm and the reference wavelength of 450 nm, and the inhibition rate was calculated: [inhibition rate (%) = (1 - experimental group OD value / control group OD group) × 100%] The experimental results of the half-inhibitory concentration (IC ₅₀ ) of each concentration were expressed as x±s, and were processed by SPSS 17.0 statistical software and analyzed by t test and one-way analysis of variance. P<0.05 was considered statistically significant.

The formula of the experimental group is the formula in the first embodiment,

The formula of Control 1 is ferul 20g 蟾蜍25g Mabao 40g Hedyotis diffusa 140g;

The formula of Control 2 is Awei 40g 蟾蜍35g Mabao 60g Hedyotis diffusa 160g;

The extraction method is referred to in Example 1.

For the blank control, only the medium is added.

The experimental results of the Lee's glutinous water extract (the formulation of Example 1) are shown in Tables 1, 2 and 3.

Table 1 "Li's glutinous food powder" (formulation of Example 1) water extract inhibited hepatoma cell HepG-2 (x ± s, n = 3)

Table 2 "Li's glutinous food" (formulation of Example 1) water extracts inhibited gastric cancer cells MKN-45 (x ± s, n = 3)

Table 3 "Li's glutinous food" (formulation of Example 1) aqueous extracts inhibited esophageal cancer cells EC-109 (x ± s, n = 3)

From the above results, it can be seen that the extract of Lee's glutinous rice water in the range of 15.62-1000 μg/ml for liver cancer cell lines (HepG-2), gastric cancer cell lines (MKN-45) and esophageal cancer cell lines (EC- 109) has a certain inhibitory effect. In contrast, the inhibitory effect on Hepatoma cell line (HepG-2) was slightly stronger, and the half-inhibitory concentration was 143.59 μg/ml. The inhibition rate of gastric cancer cell line (MKN-45) and esophageal cancer cell line (EC-109) in this concentration range was not more than 50%.

The experimental results of the alcohol extract of Lee's cockroach (Examples 1, Control 1 and Control 2) are shown in Table 4-12, respectively.

Table 4 "Li's glutinous food" ethanol extract (the formulation of Example 1) inhibited hepatoma cell HepG-2 (x ± s, n = 3)

Table 5 "Li's glutinous food" ethanol extract (the formulation of Example 1) inhibited gastric cancer cells MKN-45 (x ± s, n = 3)

Table 6 "Li's glutinous food" ethanol extract (the formulation of Example 1) inhibited esophageal cancer cells EC-109 (x ± s, n = 3)

Table 7: Inhibition of hepatoma cell HepG-2 by ethanol extract of Lee's Dietary Prescription 1 (prescription 1) (x±s, n=3)

Table 8: Inhibition of gastric cancer cell MKN-45 by ethanol extract of Lee's Dietary Prescription 1 (prescription 1) (x±s, n=3)

Table 9: Inhibition of esophageal cancer cell line EC-109 by ethanol extract of Lee's Dietary Prescription 1 (prescription 1) (x±s, n=3)

Table 10: Inhibition of hepatocellular carcinoma cell line HepG-2 by ethanol extract of Lee's Dietary Prescription 2 (prescription 2) (x±s, n=3)

Table 11: Inhibition of gastric cancer cell MKN-45 by ethanol extract of Lee's Dietary Prescription 2 (Formulation of Control 2) (x±s, n=3)

Table 12: Inhibition of esophageal cancer cell line EC-109 by ethanol extract of Lee's Dietary Prescription 2 (Formulation of Control 2) (x±s, n=3)

From the above results, it can be seen that the extract of Li Shizhen food powder in the range of 23.44-1500 μg/ml against hepatoma cell line (HepG-2), gastric cancer cell line (MKN-45) and esophageal cancer cell line (EC) -109) had a good inhibitory effect, the highest inhibition rate was about 90%, and the half-inhibitory concentrations of HepG-2, MKN-45 and EC-109 were: 622.89, 173.41 and 210.17 μg/mL, respectively. .

It also shows that when the amount of each component is slightly changed, the effect is also greatly affected, and the anticancer effect is greatly reduced (see Table 7-12). Up to 50%. The formulation of Example 1 was up to 90% compared to this, and the formulation of Example 1 was effective.

Although the embodiments of the present invention have been shown and described, it is understood that the foregoing embodiments are illustrative and not restrictive Variations, modifications, alterations and variations of the above-described embodiments are possible within the scope of the invention.

Claims

An antitumor drug characterized by being composed of the following components in a weight ratio, which is: Awei: 蟾蜍: Mabao: Hedyotis diffusa = 30:30:50:150.
The method for producing an antitumor drug according to claim 1, wherein each component in claim 1 is weighed, and each component is pulverized, sieved, and mixed.
The method for producing an antitumor drug according to claim 2, wherein each of the components of the antitumor drug obtained by the screening has a diameter of 300 to 800 mesh.
The use of the anti-tumor drug of claim 1 for anti-tumor use.
The use of the antitumor drug according to claim 4 for antitumor, characterized in that the tumor is liver cancer, gastric cancer, esophageal cancer.
The use of the antitumor drug according to claim 4 for antitumor use, comprising the step of delivering the antitumor drug to water or rice wine.