CN100493522C - A kind of medicinal composition of matrine and polysaccharide - Google Patents

Abstract

The present invention belongs to the field of medicine technology, and discloses one kind of mechanical composition, which consists of kurarione, glossy ganoderma or glossy ganoderma polysaccharide, and lentinan. It is used in treating liver cancer, lung cancer, osophagus cancer, hepatitis, etc and may be also used in the operational treatment, radiotherapy and chemotherapy of cancer. It has synergistic effect of the components, may be prepared into different forms and possesses wide application foreground.

Description

A kind of medicinal composition of matrine and polysaccharide

1. Technical field

The invention relates to a pharmaceutical composition mainly made of matrine, ganoderma lucidum or ganoderma polysaccharide and lentinan, a preparation method and application thereof, and belongs to the technical field of medicine.

2. Background technology

Cancer is a kind of disease that seriously threatens human life and health. According to statistics, about 1 million cancer patients are newly discovered in my country every year, while the world kills about 6 million people every year, and puts 10 million people on the verge of death. With the deteriorating environment of human beings, the occurrence of cancer The rate is increasing year by year. The World Health Organization predicts that cancer will become the "first killer" of human beings in the 21st century. At present, the treatment of cancer in modern medicine is mainly surgical treatment combined with radiotherapy and chemotherapy. Although surgery can remove the primary focus, it cannot fundamentally prevent the regeneration and reproduction of cancer cells; although radiotherapy and chemotherapy can kill cancer cells, they also A large number of normal tissue cells are damaged, causing gastrointestinal reactions, bone marrow suppression, liver, kidney, and heart function damage, making the patient's body weaker and difficult for further treatment. Traditional Chinese medicine has a long history of treating cancer and has formed its own unique theoretical system and treatment principles. In recent years, the work of the vast number of medical workers has proved that traditional Chinese medicine has a prominent role in the treatment of cancer, especially in the rehabilitation of cancer after surgery and in the enhancement and detoxification of radiotherapy and chemotherapy.

Matrine is a mixed base of oxymatrine and a very small amount of oxysophocarpine. It is a white powder extracted and separated from the dried root of Sophora flareescen Ait., which has been collected in State Drug Administration National Drug Standards Chemical Drugs Local Standards Increase National Standard Volume 16, Page 363 (Compiled by National Pharmacopoeia Commission), which stipulates that the content of oxymatrine (C ₁₅ H ₂₄ N ₂ O ₂ ) shall not be less than 98.0%. Matrine can reduce the content of HBsAg and HBcAg in the liver of hepatitis B virus transgenic mice, and has the effect of anti-hepatitis B virus; it can reduce the activity of liver inflammation, inhibit the synthesis of intrahepatic collagen and resist liver fibrosis; Inhibit the secretion of TNFα, block the liver toxicity of TNFα, block the apoptosis and necrosis of liver cells, etc.; it has anti-inflammatory, anti-virus, anti-liver fibrosis, liver protection and enzyme reduction, anti-cancer, white blood cell increase, anti-arrhythmia, etc. It is an adjuvant drug for tumors and liver diseases, has little toxic and side effects, and can improve immunity. It is an ideal anti-cancer and liver-protecting drug. The structural formula of matrine is as follows:

matrine

Ganoderma lucidum is the dry fruiting body of Ganoderma lucidum (Ley-ss.ex Fr.) Karst. or Ganoderma sinense Zhao, Xu et Zhang. Sweet in nature, flat, homecoming heart, lung, liver, kidney meridian. It has the effects of invigorating qi and calming the nerves, relieving cough and relieving asthma, and is mainly used for dizziness and insomnia, palpitation, shortness of breath, and cough due to fatigue. Ganoderma lucidum is a well-known traditional Chinese medicine, commonly known as "immortal grass". It was first recorded in "Shen Nong's Materia Medica". "Compendium of Materia Medica" believes that Ganoderma lucidum has the effects of "nourishing and strengthening", "prolonging life", "benefiting joints", and "curing deafness". The chemical composition of Ganoderma lucidum is complex, currently known to contain polysaccharides (peptides), triterpenoids, nucleosides, alkaloids, amino acids and trace elements. Polysaccharide compounds are one of the main chemical components contained in Ganoderma lucidum, and there are many types of polysaccharides in Ganoderma lucidum, including water-soluble polysaccharides, acidic polysaccharides and alkaline polysaccharides. It has been proven that Ganoderma lucidum polysaccharides have anti-tumor effects, liver protection and detoxification effects, immune regulation effects, blood sugar-lowering effects, blood-lipid-lowering effects, anti-oxidation effects and anti-aging effects. Clinical trials have also confirmed that Ganoderma lucidum polysaccharides can be used as an effective adjuvant drug for acute and chronic hepatitis, tumor chemotherapy and radiotherapy, and reduce the toxic and side effects caused by surgery, radiotherapy and chemotherapy. It is an ideal medicinal material for biological treatment.

Lentinan (lentinan, referred to as LNT) is currently produced by many companies in my country, and lentinan injection and lentinan for injection are listed. At present, lentinan has been used as a chemical raw material drug. The listed manufacturers include: Nanjing Kanghai Pharmaceutical Co., Ltd. ) and Nanjing Yiheng Pharmaceutical Co., Ltd. (Guoyao Zhunzi H20055744), etc. Lentinan has the functions of regulating immune function, stimulating the formation of interferon, anti-tumor and anti-virus. medicine.

At present, it has not been reported that matrine, Ganoderma lucidum or Ganoderma lucidum polysaccharide interacts with Lentinan to prepare medicines for treating cancer or hepatitis by combining prescriptions.

3. Contents of the invention

In order to meet the clinical needs, the present invention provides a new pharmaceutical composition and its preparation method. The pharmaceutical composition is mainly made of effective doses of matrine, ganoderma or ganoderma polysaccharide and lentinan for the treatment of cancer and hepatitis Aspects of the disease had unexpected effects.

The pharmaceutical composition of the present invention is prepared from the following raw materials in parts by weight: 80-2000 parts of matrine, 600-15000 parts of Ganoderma lucidum, 1-50 parts of lentinan, preferably: 200-800 parts of matrine, 1500-1500 parts of Ganoderma lucidum 6000 parts, 4-16 parts of lentinan, more preferably: 400 parts of matrine, 3000 parts of ganoderma lucidum, and 8 parts of lentinan.

Ganoderma lucidum in the above pharmaceutical composition can be prepared with suitable solvent and method to obtain Ganoderma lucidum polysaccharide, the extraction solvent is preferably water or ethanol, the extraction method can be dipping method, percolation method, decoction method, reflux extraction method or continuous extraction method, extraction The obtained Ganoderma lucidum polysaccharide is mixed with matrine, lentinan and pharmaceutically acceptable auxiliary materials to prepare any preparation. Kushensu and lentinan are chemical raw materials that have been listed in China and can be purchased directly from the market.

The invention provides a kind of preferred Ganoderma lucidum extraction process, specifically as follows:

Take Ganoderma lucidum, crush it into coarse grains, add 4 times the amount of water to moisten overnight, decoct three times the next day, add 12 times the amount of water to decoct for 3 hours for the first time, add 10 times the amount of water for 2 hours, and decoct for the second time Add 10 times the amount of water three times and decoct for 1 hour, combine the decoction, filter, concentrate the filtrate to a relative density of 1.10-1.15, filter, use the Sevage method to remove protein from the filtrate, filter, add ethanol to the filtrate until the alcohol content is 60%, stir well, refrigerate for 24 hours, filter, collect the filter cake, add appropriate amount of water to stir and dissolve, filter, add ethanol to the filtrate until the alcohol content is 85%, stir well, refrigerate for 24 hours, filter After washing, collect the filter cake, add appropriate amount of acetone and ethanol to wash repeatedly, filter with suction, add appropriate amount of water to dissolve the precipitate, ultrafilter, collect the part with a molecular weight greater than 50000 Daltons, concentrate, and dry in vacuum to obtain the product.

The yield of ganoderma lucidum extract prepared by the process is 0.5-2%, and the content of ganoderma lucidum polysaccharide is not less than 80%.

In addition to the above methods, it can also be obtained by the following methods, but not limited to the following methods:

Method 1: Take Ganoderma lucidum, crush it into coarse grains, add 3 times the amount of water to moisten overnight, add water to decoct three times the next day, add 12 times the amount of water to extract for 3 hours for the first time, add 10 times the amount of water for the second and third times to extract 2 hours, combine the decoctions, filter, concentrate the filtrate to a relative density of 1.03-1.08, filter, use the Sevage method to remove protein from the filtrate, filter, add ethanol to the filtrate to make the alcohol content 70%, stir well, and refrigerate for 24 hours , filter, collect the filter cake, add an appropriate amount of water to stir to dissolve, filter, add ethanol to the filtrate to make the alcohol content 85%, stir well, put it in refrigeration for 24 hours, filter, collect the filter cake, add appropriate amount of acetone and ethanol to wash Repeated washing several times, suction filtration, adding an appropriate amount of water to dissolve the precipitate, refrigerating for 24 hours, collecting the filtrate by filtration, concentrating, and vacuum drying to obtain the product.

The yield of the ganoderma lucidum extract prepared by the process is 2-4%, and the content of the ganoderma lucidum polysaccharide is not less than 60%.

Method 2: Take Ganoderma lucidum, crush it into coarse grains, add 3 times the amount of water to moisten overnight, decoct twice the next day, add 12 times the amount of water for the first 3 hours, add 10 times the amount of water for the second 2 hours, and combine the decoction , filtered, the filtrate was concentrated to a relative density of 1.02 to 1.06, filtered, the filtrate was deproteinized by the Sevage method, filtered, the filtrate was added with ethanol to make the alcohol content 65%, stirred evenly, refrigerated for 24 hours, filtered, collected Filter cake, add appropriate amount of water to dissolve, filter, add ethanol to the filtrate to make the alcohol content 80%, stir well, refrigerate for 24 hours, filter, collect filter cake, add appropriate amount of water to dissolve, refrigerate for 48 hours, filter After passing, the filtrate was concentrated and vacuum-dried to obtain the obtained product.

The yield of the ganoderma lucidum extract prepared by the process is 3-5%, and the content of the ganoderma lucidum polysaccharide is not less than 50%.

The pharmaceutical composition of the present invention can also be prepared by direct feeding of matrine, ganoderma lucidum polysaccharide and lentinan. According to the yield calculation of the extract relative to the medicinal materials, the weight parts of the raw material of the pharmaceutical composition of the present invention is: matrine 80-2000 3-300 parts of Ganoderma lucidum polysaccharide, 1-50 parts of Lentinan, preferably: 200-800 parts of matrine, 5-120 parts of Ganoderma polysaccharide, 4-16 parts of Lentinan, more preferably: 400 parts of matrine , Ganoderma lucidum polysaccharide 15-60 parts, mushroom polysaccharide 8 parts.

The polysaccharide content of the ganoderma lucidum polysaccharide in the above pharmaceutical composition is not less than 50%, preferably not less than 80%.

The above composition is proportioned by weight, and can be increased or decreased according to the corresponding proportion during production. For example, the unit of kilogram or ton can be used for large-scale production, and the unit of gram can also be used for small-scale production. The weight can be increased or decreased, but the weight ratio between the components remains unchanged. If the unit is gram, it can be made into 100-10000 times of preparations. If it is used as an injection, it can be made into 100-10000 pieces, and the dosage is 1-10 pieces each time. If it is used as a tablet, it can be made into 100-10000 pieces. Take 1-10 tablets each time.

The proportions of the above weight ratios are obtained through scientific screening. For special patients, the proportions of the composition can be adjusted accordingly, and the increase or decrease does not exceed 100%.

The pharmaceutical composition of the present invention can be used for the treatment of liver cancer, lung cancer, esophageal cancer, gastric cancer, nasopharyngeal cancer, rectal cancer, breast cancer, etc. Premedication for cancer surgery or combined radiotherapy and chemotherapy.

The pharmaceutical composition of the present invention can be added with one or more pharmaceutically acceptable carriers, and administered orally or parenterally to patients in need of such treatment. For oral administration, it can be made into conventional solid preparations, such as tablets, capsules, soft capsules, dispersible tablets, oral liquids, granules, chewable tablets, orally disintegrating tablets, dropping pills, sustained-release tablets, sustained-release capsules, Controlled-release tablets, controlled-release capsules, made into liquid preparations such as water or oil suspensions or other liquid preparations such as syrups; for parenteral administration, they can be made into injection solutions, water or oil suspensions, etc. Such as water injection, freeze-dried powder injection, sterile powder injection, infusion, etc. The preferred dosage form of the composition is oral preparation or injection, such as tablet, capsule, granule, powder injection, water injection, infusion and the like.

The pharmaceutical composition of the present invention can be produced by conventional methods in the existing pharmaceutical field, and various pharmaceutically acceptable carriers can be added when necessary. The carrier includes conventional excipients, fillers, binders, wetting agents, disintegrants, absorption promoters, surfactants, adsorption carriers, lubricants and the like in the pharmaceutical field.

When the pharmaceutical composition of the present invention is made into an oral preparation, optional fillers include: starch, powdered sugar, calcium phosphate, calcium sulfate dihydrate, dextrin, microcrystalline cellulose, lactose, pregelatinized starch, mannitol etc.; optional binders are: sodium carboxymethyl cellulose, PVP-K30, hydroxypropyl cellulose, starch slurry, methyl cellulose, ethyl cellulose, hypromellose, gelatinized starch, etc. ; Optional disintegrants are: dry starch, crospovidone, croscarmellose sodium, carboxymethyl starch sodium, low-substituted hydroxypropyl cellulose, etc.; optional lubricants are: hard Magnesium fatty acid, talc, sodium lauryl sulfate, micronized silica gel, etc.

When the pharmaceutical composition of the present invention is made into an injection, in order to increase its solubility, a solubilizer such as polysorbate 80 can be added. Isotonic regulators for adjusting osmotic pressure can be added to the infusion, for example, sodium chloride, potassium chloride, magnesium chloride, calcium chloride, sodium lactate, glucose, xylitol, sorbitol and dextran, etc., preferably sodium chloride or glucose. Excipients such as mannitol and glucose can be added to the powder for injection.

The pharmaceutical composition of the present invention has the following advantages:

(1) Provided for the first time a pharmaceutical composition that is compatible with matrine, ganoderma lucidum or ganoderma polysaccharide and lentinan for the treatment of tumors and hepatitis, which meets the urgent clinical needs;

(2) Pharmacological studies have been carried out on the interaction and compatible prescription of the pharmaceutical composition of the present invention, and it has been found that the pharmaceutical composition has a significant inhibitory effect on the growth of mouse _S180 tumors, and the tumor inhibition rate can reach more than 50%. , can significantly prolong the survival days of ascites cancer U ₁₄ mice, and the rate of life extension is also significantly increased, which can not only significantly enhance the effect of radiotherapy, but also significantly enhance the curative effect of chemotherapy, and can significantly reduce the toxicity of _chemotherapy . The mouse liver injury has obvious protective effect; and compared with matrine alone, Ganoderma lucidum or Ganoderma lucidum polysaccharide, Lentinan, it is applied to anti-tumor and liver injury protection after compatibility, and the curative effect is remarkable, and its result is common in this technical field unexpected by technicians;

(3) The preparation process of the pharmaceutical composition of the present invention is simple, the quality difference between different batches of medicines is small, and the quality of the medicines is uniform and stable;

(4) the stability experiment that carries out shows that every index of pharmaceutical composition injection of the present invention is all relatively stable, has guaranteed the safety of clinical medication;

(5) The combined medication of the composition of the present invention has definite curative effect, reduces the relative dosage, and has broad application prospects.

The beneficial effect of the pharmaceutical composition of the present invention will be further illustrated by test examples below. In the following test examples: the composition of matrine, ganoderma or ganoderma polysaccharide and lentinan is hereinafter referred to as KLX composition . The Ganoderma lucidum polysaccharides used in the following test examples are all taken from Example 1.

Test example 1 KLX composition is to mouse S ₁₈₀ tumor growth inhibitory effect

Test product: blank control group: 0.9% normal saline injection, commercially available;

Matrine group: matrine injection, specification 2ml: 0.2g;

Ganoderma lucidum polysaccharide group: Ganoderma lucidum polysaccharide injection, self-made, 2ml: 28.8mg (equivalent to 3g of Ganoderma lucidum);

Lentinan group: Lentinan injection, homemade, 2ml: 4mg;

KLX injection group: self-made (see Example 3 for the preparation method).

Test animals: 130 healthy mice, weighing 16-20 g, male and female, 10 in each group.

Tumor strain: mouse S ₁₈₀ sarcoma

Test method: Take the S ₁₈₀ tumor of the inoculated and passaged mouse, add normal saline to the homogenizer to make a homogenate of the mouse S ₁₈₀ tumor, then dilute it with normal saline 1:3, and then take 0.2ml and inject it into the left armpit of the mouse Subcutaneously, weighed for 24 hours, intraperitoneally injected into mice once a day, with the same administration volume (0.5ml/mouse), for 7 days in total. The mice were sacrificed the day after drug withdrawal, weighed and the subcutaneous tumor mass was carefully peeled off, the tumor weight was weighed on an EM50 electronic balance, and the tumor inhibition rate was calculated.

Table 1 Inhibitory effect of KLX composition injection on mouse S ₁₈₀ tumor growth (x±s, n=10)

Note: Compared with blank control group, ^* p<0.05, ^** p<0.01; compared with matrine group, ^a p<0.05; compared with Ganoderma lucidum polysaccharide group, ^b p<0.05 Compared with lentinan group , ^cp <0.05.

Test results: see Table 1.

(1) Compared with the blank control group, the matrine group, the ganoderma lucidum polysaccharide group and the lentinan group could significantly inhibit the growth of mouse S ₁₈₀ sarcoma (p<0.05); Growth of S ₁₈₀ sarcoma in mice (p<0.01).

(2) Compared with matrine group, each proportioning group of KLX composition can significantly inhibit the growth of mouse S ₁₈₀ sarcoma (p<0.05); compared with Ganoderma lucidum polysaccharide group, each proportioning group of KLX composition It can significantly inhibit the growth of S ₁₈₀ sarcoma in mice (p<0.05); compared with the lentinan group, each proportioning group of KLX composition can significantly inhibit the growth of S ₁₈₀ sarcoma in mice (p<0.05).

Conclusion: Compared with matrine, ganoderma polysaccharide, and lentinan alone, the combination of matrine, ganoderma polysaccharide and lentinan can significantly inhibit the growth of S ₁₈₀ sarcoma in mice, and significantly increase the tumor inhibition rate of mice. The tumor inhibition rate reached more than 50%. The results show that in the range of matrine (200mg-800mg) + Ganoderma lucidum (1.5g-6g) + lentinan (4mg-16mg), the anticancer effect of the composition is relatively satisfactory, and the ratio is matrine + Ganoderma lucidum +Leninan=400mg+3g+8mg is the best. It is suggested that matrine has a synergistic effect when combined with Ganoderma lucidum polysaccharide and Lentinan.

Test example 2 KLX composition on ascites cancer U ₁₄ Effects on the rate of life extension in mice

Test product: blank control group: 0.9% normal saline injection, commercially available;

Matrine group: matrine injection, specification 2ml: 0.2g;

Ganoderma lucidum polysaccharide group: Ganoderma lucidum polysaccharide injection, self-made, 2ml: 28.8mg (equivalent to 3g of Ganoderma lucidum);

Lentinan group: Lentinan injection, homemade, 2ml: 4mg;

KLX injection group: self-made (refer to Example 3 for prescription and preparation method), divided into high, middle and low dosage groups.

Test animals: 70 healthy mice, weighing 20-25 g, both male and female, randomly divided into 7 groups, 10 mice in each group.

Tumor strain: mouse ascites carcinoma U ₁₄

Test method: Mice were inoculated with ascites cancer U ₁₄ tumor strain bacterial suspension (suspension concentration 2×10 ⁷ /ml, inoculation volume 0.5ml/mouse). On the second day after inoculation, the mice were randomly divided into groups, weighed, and administered intraperitoneally according to Table 2, once a day for 10 consecutive days. Afterwards, the death time of the mice was observed, and the results were expressed by average survival days and life extension rate [life extension rate=(average survival days of the test group-average survival days of the control group)/average survival days of the control group×100%].

Table 2 Effect of KLX composition injection on life extension rate of ascites cancer _U14 mice (x±s, n=10)

Note: Compared with blank control group, ^** p<0.01, ^*** p<0.001; compared with matrine group, ^a p<0.05, ^aa p<0.01; compared with Ganoderma lucidum polysaccharide group, ^b p< 0.05, ^bb p<0.01; compared with lentinan group, ^c p<0.05, ^cc p<0.01.

Test results: see Table 2.

(1) Compared with the blank control group, the matrine group, the ganoderma lucidum polysaccharide group and the lentinan group can significantly prolong the survival days of ascites cancer U ₁₄ mice (p<0.01); Significantly prolong the survival days of ascites carcinoma U ₁₄ mice (p<0.001).

(2) Compared with the matrine group, the low and middle dose groups of KLX composition can significantly prolong the survival days of U ₁₄ mice with ascites cancer (p<0.05), and the high dose group of KLX composition can significantly prolong the survival time of U ₁₄ mice with ascites cancer. The survival days of mice (p<0.01); compared with the Ganoderma lucidum polysaccharide group, the KLX composition low and middle dose groups can significantly prolong the survival days of ascites cancer U ₁₄ mice (p<0.05), and the KLX composition high dose group Can significantly prolong the survival days of ascites cancer U ₁₄ mice (p<0.01); compared with the lentinan group, the KLX composition low and medium dose groups can significantly prolong the survival days of ascites cancer U ₁₄ mice (p<0.05) , the high-dose KLX composition group can significantly prolong the survival days of ascites cancer U ₁₄ mice (p<0.01).

Conclusion: Compared with matrine, ganoderma polysaccharide, and lentinan alone, the combination of matrine, ganoderma polysaccharide, and lentinan can prolong the survival days of ascites cancer U ₁₄ mice, and the life extension rate is also significantly improved; and The effect is related to the dosage, and the effect is the best when the dosage is high. It is suggested that matrine has a synergistic effect when combined with Ganoderma lucidum polysaccharide and Lentinan.

Test example 3 The synergistic effect of KLX composition on radiotherapy

Test product: blank control group: 0.9% normal saline injection, commercially available;

⁶⁰ Co irradiation group: ⁶⁰ Co irradiation;

Matrine group: matrine injection, specification 2ml: 0.2g;

KLX injection group: self-made (refer to Example 3 for prescription and preparation method), divided into high, middle and low dosage groups.

Test animals: 60 healthy mice, weighing 20-25 g, both male and female, randomly divided into 6 groups, 10 in each group.

Tumor strain: mouse S ₁₈₀ sarcoma.

Test method: each mouse was subcutaneously inoculated with S ₁₈₀ tumor cell suspension (suspension concentration 2×10 ⁷ /ml, inoculation volume 0.2ml/mouse) under the armpit of the left forelimb of each mouse, and weighed after 24 hours. On the second day after inoculation, they were randomly divided into groups. Except for the blank control group, the rest of the groups were irradiated with ⁶⁰ Co whole body on the 3rd and 6th days after inoculation, and the irradiation dose was 0.05Gy/min. On the next day of inoculation, the mice were administered intraperitoneally according to Table 3, once a day for 10 consecutive days. The body weight was weighed every day, and the body weight change of the tumor-bearing mice was observed. 24 hours after the last administration, weigh the body, kill the animal, peel off the subcutaneous tumor mass, weigh the tumor body weight, calculate the tumor inhibition rate and the increase rate [increase rate=(average tumor weight of the radiotherapy group-radiotherapy and composition injection combined treatment) group average tumor weight)/radiotherapy group average tumor weight×100%].

Table 3 The synergistic effect of KLX composition on radiotherapy (x±s, n=10)

Note: Compared ^with blank control group, ^* p<0.05, ^** p<0.01, ^*** p<0.001; compared with ⁶⁰ Co irradiation group, ^a p<0.05, ^aa p<0.01; Compared with the plain group, ^b p<0.05.

Test results: see Table 3.

(1) Compared with the blank control group, ^{the 60} Co irradiation group had a significant inhibitory effect on the mouse S ₁₈₀ sarcoma (p<0.05); the combined treatment of ⁶⁰ Co irradiation and matrine had a significant inhibitory effect on the mouse S ₁₈₀ sarcoma Effect (p<0.01); The combined treatment of ⁶⁰ Co irradiation and each dose group of KLX composition has a very significant inhibitory effect on mouse S ₁₈₀ sarcoma (p<0.001).

(2) Compared with the ⁶⁰ Co irradiation group, the combined treatment of ⁶⁰ Co irradiation and matrine group had significantly enhanced inhibitory effect on mouse S ₁₈₀ sarcoma (p<0.05), and the tumor inhibition rate was significantly increased (p<0.05); ⁶⁰ The combined treatment of Co irradiation and each dose group of KLX composition significantly enhanced the inhibitory effect on mouse S ₁₈₀ sarcoma (p<0.01), and the tumor inhibition rate was significantly increased (p<0.01).

(3) Compared with the ⁶⁰ Co irradiation + matrine group, the combined treatment of ⁶⁰ Co irradiation and KLX composition dosage groups significantly enhanced the inhibitory effect on mouse S ₁₈₀ sarcoma (p<0.05), and the tumor inhibition rate was significantly increased ( p<0.05).

Conclusion: Compared with ⁶⁰ Co irradiation alone and matrine combined application group, each dose group of KLX composition can significantly enhance the curative effect of radiotherapy, and the combination of matrine, Ganoderma lucidum polysaccharide and lentinan combined with ⁶⁰ Co irradiation combined treatment of cancer has a significant effect. A satisfactory therapeutic effect has been achieved.

Experimental example 4 The synergistic and attenuating effect of KLX composition on chemotherapy

Test product: blank control group: 0.9% normal saline injection, commercially available;

CTX control group: cyclophosphamide injection, self-made;

Matrine group: matrine injection, specification 2ml: 0.2g;

KLX injection group: self-made (refer to Example 3 for prescription and preparation method), divided into high, middle and low dosage groups.

Test animals: 60 healthy mice, weighing 20-25 g, both male and female, randomly divided into 6 groups, 10 in each group.

Tumor strain: mouse S ₁₈₀ sarcoma.

Test method: each mouse was subcutaneously inoculated with S ₁₈₀ tumor cell suspension (suspension concentration 2×10 ⁷ /ml, inoculation volume 0.2ml/mouse) under the armpit of the left forelimb. On the second day after inoculation, the mice were randomly divided into groups, weighed, and administered intraperitoneally according to the dosage in Table 4, once every other day, for 10 consecutive days. The body weight was weighed every day, and the body weight change of the tumor-bearing mice was observed. 24 hours after the last administration, the body weight was weighed, the animals were sacrificed, the subcutaneous tumor mass was peeled off, the tumor body weight was taken, and the tumor inhibition rate and enhancement rate were calculated [increase rate=(average tumor weight of chemotherapy group-chemotherapy and KLX composition combined drug group) (average tumor weight)/average tumor weight in chemotherapy group×100%]; dissect thymus, spleen and target organs, weigh and calculate organ index.

Table 4 The synergistic and attenuating effects of KLX composition on chemotherapy (x±s, n=10)

Note: Compared with blank control group, ^* p<0.05, ^** p<0.001, ^*** p<0.01; compared with CTX group, ^a p<0.05, ^aa p<0.01; compared with CTX+matrine group Ratio, ^b p<0.05, ^bb p<0.01.

Test results: see Table 4.

(1) Synergistic effect on radiotherapy:

Compared with the blank control group, the CTX group had a significant inhibitory effect on mouse S ₁₈₀ sarcoma (p<0.05); the combination of CTX and matrine had a significant inhibitory effect on mouse S ₁₈₀ sarcoma (p<0.05); The combined administration of CTX and KLX in each dose group has a very significant inhibitory effect on mouse S ₁₈₀ sarcoma (p<0.001).

Compared with the CTX control group, the combination of CTX and matrine had a significant inhibitory effect on mouse S ₁₈₀ sarcoma (p<0.05), and the tumor inhibition rate was significantly increased (p<0.05); the combination of CTX and KLX in each dose group It has a significant inhibitory effect on mouse S ₁₈₀ sarcoma (p<0.01), and the tumor inhibition rate is also significantly increased (p<0.01).

Compared with the CTX+matrine group, the combination of CTX and KLX low and medium dose groups had a significant inhibitory effect on mouse S ₁₈₀ sarcoma (p<0.05), and the tumor inhibition rate was also significantly increased (p<0.05); The combination of KLX high-dose group had a significant inhibitory effect on mouse S ₁₈₀ sarcoma (p<0.01), and the tumor inhibition rate was significantly increased (p<0.01).

(2) Attenuating effect on chemotherapy:

Compared with the blank control group, when cyclophosphamide was administered alone, the number of peripheral leukocytes, thymus index, and spleen index of the mice were all significantly decreased (p<0.001); compared with the CTX control group, the combination of CTX and matrine It can significantly inhibit the reduction of peripheral leukocyte count, thymus index, and spleen index in mice; the combination of CTX and KLX dosage groups can significantly inhibit the decrease of peripheral leukocyte count, thymus index, and spleen index in mice; In comparison, the combination of CTX and KLX medium dose group can significantly inhibit the decrease of peripheral leukocyte count, thymus index and spleen index of mice, and the combination of CTX and KLX high dose group can significantly inhibit the peripheral leukocyte count, thymus index, spleen index of mice. Decreased spleen index.

Conclusion: Compared with the single application group of CTX and matrine, the combination of CTX and KLX can significantly enhance the efficacy of chemotherapy, and can significantly reduce the toxicity of chemotherapy. It is suggested that matrine, ganoderma lucidum polysaccharide and lentinan can be used as an adjuvant drug for chemotherapy, which can increase efficacy and reduce toxicity, and the therapeutic effect is satisfactory.

Test example 5 KLX composition is to carbon tetrachloride (CCl ₄ )-induced acute liver injury in mice

Test product: chemically pure CCl ₄ , Chengdu United Chemical Reagent Research Institute;

Blank control group: 0.9% normal saline, self-made;

Matrine group: matrine injection, specification 2ml: 0.2g;

KLX injection group: self-made (refer to Example 3 for prescription and preparation method), divided into high, middle and low dosage groups.

Experimental animals: ICR mice, weighing 22-27 g, half male and half male.

Experimental method: 60 mice were taken and randomly divided into 6 groups, namely blank control group, model group, matrine group, and KLX composition dosage groups, with 10 mice in each group. The blank control group and the model group were injected with normal saline 20ml/kg daily, once a day, for 7 consecutive days; the administration group was given tail vein injection, once a day, with the dosage shown in the table below, for 7 consecutive days. The blank control group was intraperitoneally injected with peanut oil 10ml/kg 2 hours after the last tail vein injection, and the animals in the other groups were intraperitoneally injected with 0.12% CCl ₄ peanut oil solution 10ml/kg. After 16 hours, the animals were killed by decapitation, and the serum was taken to test the values of ALT and AST. After decapitation, the liver and spleen were immediately taken out by laparotomy, the blood was sucked, the fat and mesangium were cut off, the weight of the liver and spleen was accurately weighed, and the liver index and spleen index were calculated. The results are shown in Table 5.

Table 5 Effect of KLX composition on liver and spleen index and serum ALT and AST levels in mice with acute liver injury caused by _CCl4

Note: Compared with blank control group, ^s p<0.05, ^ss p<0.01, ^sss p<0.001; compared with model group, ^* p<0.05, ^** p<0.01; compared with matrine group, ^# p<0.05 .

Experimental results: see Table 5.

(1) Compared with the blank control group, the liver index of the mice in the CCl ₄ model group was significantly increased (p<0.01), the spleen index was significantly increased (p<0.05), and the serum ALT and AST values were significantly increased (p <0.001), indicating that mice were injected with CCl ₄ peanut oil solution for liver injury, and the modeling was successful.

(2) Compared with the model group, the liver index of the mice in the matrine group was significantly lower (p<0.05), the spleen index had no statistical significance, and the serum ALT and AST values were significantly lower (p<0.05). The mouse liver index, serum ALT, AST value of each dose group of KLX composition significantly reduces (p<0.01), and the mouse spleen index significantly reduces (p<0.05), illustrates that each administration group is to CCl ₄ induced acute liver injury. Has a protective effect.

(3) Compared with the matrine group, the liver index, spleen index, ALT, and AST values of the mice in each dose group of the KLX composition were significantly decreased (p<0.05).

Conclusion: Compared with matrine alone, the combination of matrine, ganoderma lucidum polysaccharide and lentinan can significantly inhibit the increase of liver index, spleen index, serum ALT and AST values of mice induced by CCl ₄ . The effect is good. It is suggested that matrine is compatible with Ganoderma lucidum polysaccharide and Lentinan, synergistically.

Test example 6 KLX composition stability experiment

Test product: KLX injection group: self-made (prescription and preparation method refer to the preparation of embodiment 3 water injection)

Inspection items: traits, pH value, clarity, related substances, content.

Long-term stability test methods and results: each composition of this product was placed under the conditions of temperature 25°C±2°C and relative humidity 60%±10% for 6 months and 12 months, and there was no significant change in each index. It shows that the long-term storage of the pharmaceutical composition of the present invention is basically stable.

4. Specific implementation

The above-mentioned content of the present invention will be further described in detail through specific implementation in the form of examples below. However, it should not be construed that the scope of the above-mentioned subject matter of the present invention is limited to the following examples. All technologies realized based on the above contents of the present invention belong to the scope of the present invention. The auxiliary materials of each dosage form in the following examples can be replaced with pharmaceutically acceptable auxiliary materials, or decreased or increased. The Ganoderma lucidum polysaccharides in the following Examples 2-9 are taken from Example 1.

Embodiment 1 Preparation of Ganoderma lucidum polysaccharide

Take Ganoderma lucidum, crush it into coarse grains, add 4 times the amount of water to moisten overnight, decoct three times the next day, add 12 times the amount of water to decoct for 3 hours for the first time, add 10 times the amount of water for 2 hours, and decoct for the second time Add 10 times the amount of water three times and decoct for 1 hour, combine the decoction, filter, concentrate the filtrate to a relative density of 1.10-1.15, filter, use the Sevage method to remove protein from the filtrate, filter, add ethanol to the filtrate until the alcohol content is 60%, stir well, refrigerate for 24 hours, filter, collect the filter cake, add appropriate amount of water to stir and dissolve, filter, add ethanol to the filtrate until the alcohol content is 85%, stir well, refrigerate for 24 hours, filter After washing, collect the filter cake, add appropriate amount of acetone and ethanol to wash repeatedly, filter with suction, add appropriate amount of water to dissolve the precipitate, ultrafilter, collect the part with a molecular weight greater than 50000 Daltons, concentrate, and dry in vacuum to obtain the product.

Identification of Ganoderma lucidum polysaccharide

Take 50 mg of this product, add 25 ml of ethanol, heat to reflux for 30 minutes, filter, evaporate the filtrate to dryness, add 2 ml of methanol to the residue to dissolve, and use it as the test solution. Another 2 g of Ganoderma lucidum reference medicinal material was taken, and the reference medicinal material solution was prepared in the same way. According to the thin-layer chromatography test, draw 5ul of each of the above two solutions, and place them on the same silica gel G plate respectively. Developing agent, develop, take out, dry, and inspect under ultraviolet light (365nm). In the chromatogram of the test product, at the position corresponding to the chromatogram of the control medicinal material, there are fluorescent spots of the same color.

Determination of Ganoderma lucidum polysaccharide content

Preparation of reference substance solution Accurately weigh an appropriate amount of glucose reference substance dried at 105°C to constant weight, add water to make a solution containing 0.1mg per 1ml, and obtain it.

Preparation of the standard curve Accurately draw the reference substance solution 0.2ml, 0.4ml, 0.6ml, 0.8ml, 1.0ml, 1.2ml respectively, put it in a 10ml stoppered test tube, add water to 2.0ml, add anthrone sulfate solution precisely (accurately weighed Anthrone 0.1g, add 100ml of 80% sulfuric acid solution to dissolve, shake well) 6ml, shake well, heat in water bath for 15 minutes, take out, put in water bath and cool for 15 minutes, use corresponding solvent as blank, measure at 625nm Absorbance, with absorbance as the ordinate and concentration as the abscissa, draw a standard curve.

Preparation of the test solution Take 20mg of this product, put it in a 50ml measuring bottle, add water to dissolve, dilute to the mark, and shake well.

Determination method Precisely draw 2ml of the test solution, put it in a 10ml test tube, measure the absorbance according to the method under the standard curve, read the weight of the glucose in the test solution from the standard curve, and calculate it.

Three batches of Ganoderma lucidum polysaccharides were prepared by this process, and the content and yield are shown in the table below. It can be seen from the results that the yield of Ganoderma lucidum polysaccharide prepared by this process is 0.5-2%, and the polysaccharide content is not less than 80%.

Table 6 Determination results and yields of polysaccharide content of three batches of Ganoderma lucidum polysaccharide

The preparation of embodiment 2 KLX composition powder injection

1. Prescription:

2. Specific steps:

1) Firstly, aseptically process the containers used for liquid preparation, antibiotic glass bottles, rubber stoppers, etc.

2) Weigh the raw materials and auxiliary materials according to the prescription quantity.

3) Add appropriate amount of water for injection to matrine and mannitol, heat and stir to dissolve, take 70% sterile water for injection, add polysorbate 80, heat and stir to dissolve, then add the prescribed amount of Ganoderma lucidum polysaccharide and shiitake mushroom Polysaccharide, stir to dissolve completely. Combine the above solutions and add sterile water for injection to 5000ml.

4) Add activated carbon for needles with a dosing volume of 0.1%, heat and stir for 15 minutes.

5) Decarbonize by filtering through a sand filter rod, measure and adjust the pH value of the solution.

6) Fine filtration through a 0.22 μm microporous membrane.

7) Check the clarity of the solution and test the semi-finished product.

8) Divide into antibiotic glass vials and stopper halfway. Place the samples in a lyophilizer for freeze drying. Pre-freeze at -45°C for 6 hours, vacuum dry at low temperature at -45°C to 0°C for 24 hours, then heat up to 30°C and dry in vacuum for 5 hours.

9) After the freeze-drying is completed, press the stopper and roll the cap.

10) Full inspection of finished products, packaging and storage.

The preparation of embodiment 3 KLX composition aqueous injection

1. Prescription:

2. Specific steps:

1) Dispose of the pipes and containers used for liquid preparation one day in advance, and rinse them with fresh water for injection before use.

2) Add an appropriate amount of water for injection of matrine, heat and stir to dissolve, take 70% sterile water for injection, add polysorbate 80, heat and stir to dissolve, then add the prescribed amount of ganoderma lucidum polysaccharide and lentinan, and stir Dissolve completely. Combine the above solutions and add sterile water for injection to 5000ml.

3) Add activated carbon for needles with a dosing volume of 0.1%, heat and stir for 15 minutes.

4) Decarbonize by filtering through a sand filter rod, measure and adjust the pH value of the solution.

5) Fine filtration through a 0.45 μm microporous membrane.

6) Check the clarity of the solution and test the semi-finished product.

7) Seal the solution in a glass ampoule.

8) Sterilize with circulating steam at 100°C for 30 minutes.

9) Put the sample into 0.01% methylene blue solution to check for leaks while it is hot.

10) Light inspection, full inspection of finished products, packaging and storage.

The preparation of embodiment 4 KLX composition sodium chloride infusion

1. Prescription:

2. Specific steps:

1) Dispose of the pipes and containers used for liquid preparation one day in advance, and rinse them with fresh water for injection before use.

2) Add an appropriate amount of water for injection of matrine, heat and stir to dissolve, take 70% sterile water for injection, add polysorbate 80, heat and stir to dissolve, then add the prescribed amount of ganoderma lucidum polysaccharide and lentinan, and stir Dissolve completely. Dissolve sodium chloride completely with an appropriate amount of water for injection, combine the above solutions, and add water for injection to the full amount.

3) Add activated carbon for needles with a dosing volume of 0.1%, heat and stir for 15 minutes.

4) Decarburization by sand filter rod filtration. Measure and adjust the pH of the solution.

5) Fine filtration through a 0.45 μm microporous membrane.

6) Check the clarity of the solution and test the semi-finished product.

7) Fill in a 100ml infusion bottle.

8) Autoclave at 115°C for 30 minutes.

9) Light inspection, full inspection of finished products, packaging and storage.

The preparation of embodiment 5 KLX composition glucose infusion

1. Prescription:

2. Specific steps:

1) Dispose of the pipes and containers used for liquid preparation one day in advance, and rinse them with fresh water for injection before use.

2) Add an appropriate amount of water for injection of matrine, heat and stir to dissolve, take 70% sterile water for injection, add polysorbate 80, heat and stir to dissolve, then add the prescribed amount of ganoderma lucidum polysaccharide and lentinan, and stir Dissolve completely. Dissolve the glucose completely with an appropriate amount of water for injection, heat and boil for 15 minutes. Combine the above solutions and add water for injection to the full amount.

3) Add activated carbon for needles with a dosing volume of 0.1%, heat and stir for 15 minutes.

4) Decarburization by sand filter rod filtration. Measure and adjust the pH of the solution.

5) Fine filtration through a 0.45 μm microporous membrane.

6) Check the clarity of the solution and test the semi-finished product.

7) Fill in a 100ml infusion bottle.

8) Autoclave at 115°C for 30 minutes.

9) Light inspection, full inspection of finished products, packaging and storage.

The preparation of embodiment 6 KLX composition tablet

1. Prescription:

2. Specific steps:

1) Grind matrine, ganoderma polysaccharide, and lentinan, and pass through a 100-mesh sieve for later use.

2) Weigh the raw materials and auxiliary materials according to the prescription quantity.

3) Dissolving hypromellose in water to make a 2% aqueous solution for later use.

4) Mix matrine, ganoderma polysaccharide, lentinan, pregelatinized starch, and microcrystalline cellulose evenly, add an appropriate amount of 2% HPMC aqueous solution, and stir evenly to make a suitable soft material.

5) Pass through a 20-mesh sieve to make granules.

6) The particles are dried at 60°C.

7) Add magnesium stearate and sodium starch glycolate to the dried granules, pass through a 18-mesh sieve for granulation, and mix well.

8) Sampling and testing of semi-finished products.

9) Compress the tablet according to the tablet weight determined by the assay.

10) Full inspection of finished products, packaging and storage.

The preparation of embodiment 7 KLX composition capsules

1. Prescription:

2. Specific steps:

1) Grind matrine, ganoderma polysaccharide, and lentinan through a 100-mesh sieve for later use.

2) Weigh the raw materials and auxiliary materials according to the prescription quantity.

3) Dissolving hypromellose in water to make a 2% aqueous solution for later use.

4) Mix matrine, ganoderma polysaccharide, lentinan, pregelatinized starch, and microcrystalline cellulose evenly, add an appropriate amount of 2% HPMC aqueous solution, and stir evenly to make a suitable soft material.

5) Pass through a 20-mesh sieve to make granules.

6) The particles are dried at 60°C.

7) The dried granules are added with magnesium stearate, passed through a 18-mesh sieve for granulation, and mixed evenly.

8) Sampling and testing of semi-finished products.

9) Pack the capsules according to the loading determined by the assay.

10) Full inspection of finished products, packaging and storage.

The preparation of embodiment 8 KLX composition granules

1. Prescription:

2. Specific steps:

1) Grind matrine, ganoderma lucidum polysaccharide, lentinan, and sucrose through a 100-mesh sieve for later use.

2) Weigh the raw materials and auxiliary materials according to the prescription quantity.

3) Mix matrine, ganoderma lucidum polysaccharide, lentinan, sugar powder, cyclamate, citric acid, and solid essence in equal increments, add an appropriate amount of 2% HPMC50% ethanol solution, and stir evenly to make a suitable soft drink. material.

4) 20-mesh sieve to make granules.

5) The particles are dried at 60°C.

6) The particles are passed through a 18-mesh sieve for granulation.

7) Take samples, test the content of the main drug in the semi-finished product, and determine the loading.

8) Packaging, full inspection of finished products, packaging and storage.

The preparation of embodiment 9 KLX composition oral liquid

1. Prescription:

2. Specific steps:

1) Add an appropriate amount of water for injection to matrine, stir to dissolve, add an appropriate amount of ganoderma lucidum polysaccharide and lentinan to water for injection, heat and stir to dissolve completely, and combine the above solutions.

2) Sodium benzoate and stevioside are completely dissolved in water with a dosing volume of 20%.

3) Combine the above solutions and add water to the full amount.

4) Filter through a 0.8um microporous membrane.

5) Semi-finished product testing.

6) Filling.

7) Full inspection of finished products, packaging and storage.

Claims (10)

1. a pharmaceutical composition for the treatment of cancer is characterized in that, this pharmaceutical composition is made by following bulk drugs: 80～2000 parts of kurarinones, 600～15000 parts of Ganodermas, 1～50 part of lentinan.

2. pharmaceutical composition as claimed in claim 1 is characterized in that, the parts by weight of this pharmaceutical composition are: 200～800 parts of kurarinones, 1500～6000 parts of Ganodermas, 4～16 parts of lentinan.

3. pharmaceutical composition as claimed in claim 2 is characterized in that, the parts by weight of this pharmaceutical composition are: 400 parts of kurarinones, 3000 parts of Ganodermas, 8 parts of lentinan.

4. as each described preparation of drug combination method of claim 1～3, it is characterized in that, described Ganoderma prepares ganoderan with The suitable solvent and method, and ganoderan is made arbitrary preparation with kurarinone and lentinan and mixing acceptable accessories again; The content of polysaccharide is not less than 50% in the gained ganoderan.

5. pharmaceutical composition as claimed in claim 1, it is characterized in that, this pharmaceutical composition is made by following bulk drugs: kurarinone, ganoderan and lentinan, its parts by weight are: 80～2000 parts of kurarinones, 3～300 parts of ganoderans, 1～50 part of lentinan.

6. pharmaceutical composition as claimed in claim 5 is characterized in that, the parts by weight of this pharmaceutical composition crude drug are: 200～800 parts of kurarinones, 5～120 parts of ganoderans, 4～16 parts of lentinan.

7. pharmaceutical composition as claimed in claim 6 is characterized in that, the parts by weight of this pharmaceutical composition crude drug are: 400 parts of kurarinones, 15～60 parts of ganoderans, 8 parts of lentinan.

8. as each described pharmaceutical composition of claim 5～7, it is characterized in that the content of polysaccharide is not less than 50% in the described ganoderan.

9. as claim 1～3,5～7 each described pharmaceutical compositions, it is characterized in that this pharmaceutical composition and mixing acceptable accessories are made clinically any or pharmaceutically acceptable dosage form.

10. pharmaceutical composition as claimed in claim 9 is characterized in that this pharmaceutical composition is made oral formulations or injection.