阅读说明：本技术 一种代谢检查点fbp1激动剂及在抗肿瘤方面的应用 (Metabolism check point FBP1 agonist and application thereof in anti-tumor aspect ) 是由 李砺锋 张毅 赵杰 李春伟 沈志博 薛文华 于 2019-11-28 设计创作，主要内容包括：本发明公开了一种代谢检查点FBP1激动剂及在抗肿瘤方面的应用。FBP1是葡萄糖异生的关键酶之一,其通过催化1,6-二磷酸果糖水解在糖降解过程中发挥抑制作用。已知FBP1在多种肿瘤细胞中低表达,与肿瘤细胞的生长增殖相关,激活FBP1可以抑制肿瘤细胞的生长增殖。本发明提供了一种代谢检查点FBP1激活剂,该激动剂为一种天然植物多糖,CCK-8试验证明其可以显著抑制人肺腺癌、肝癌和乳腺癌细胞增殖,且呈现明显的剂量依赖性；Western blotting试验证明该天然植物多糖可以有效激活上述肿瘤细胞中FBP1蛋白表达,这可能是其发挥抗人肺腺癌、肝癌和乳腺癌的作用机制。(The invention discloses a metabolic checkpoint FBP1 agonist and application thereof in the aspect of tumor resistance. FBP1 is one of the key enzymes of gluconeogenesis, which plays an inhibitory role in the sugar degradation process by catalyzing the hydrolysis of fructose-1, 6-diphosphate. FBP1 is known to be low expressed in various tumor cells, and is associated with the growth and proliferation of tumor cells, and the growth and proliferation of tumor cells can be inhibited by activating FBP 1. The invention provides an FBP1 activator as a metabolic checkpoint, wherein the activator is a natural plant polysaccharide, and CCK-8 tests prove that the activator can obviously inhibit the proliferation of human lung adenocarcinoma, liver cancer and breast cancer cells and presents obvious dose dependence; western blotting test proves that the natural plant polysaccharide can effectively activate the expression of FBP1 protein in the tumor cells, which is probably an action mechanism for resisting human lung adenocarcinoma, liver cancer and breast cancer.)

1. A metabolic checkpoint FBP1 agonist characterized by: the polysaccharide of the golden sargentgloryvine stem is prepared by the following method:

pulverizing and sieving the lygodium japonicum, taking powder, adding deionized water, carrying out hot reflux extraction, filtering an extracting solution, carrying out reduced pressure concentration, centrifuging, collecting supernatant, adding 3 times of volume of absolute ethyl alcohol, uniformly stirring, standing for alcohol precipitation, collecting precipitate, redissolving with deionized water, centrifuging, collecting supernatant, adding 3 times of volume of absolute ethyl alcohol, uniformly stirring, standing for alcohol precipitation, collecting precipitate, washing, redissolving with deionized water, deproteinizing by a Sevag method, and freeze-drying.

2. The medical use of the sargentgloryvine stem polysaccharide of claim 1 in preparing an antitumor drug.

3. Use according to claim 2, characterized in that: the tumor is lung adenocarcinoma, liver cancer or breast cancer.

4. A pharmaceutical preparation for treating lung adenocarcinoma, which is characterized in that: the active component is a polysaccharide of the golden sargentgloryvine stem, and the golden sargentgloryvine stem is prepared into a pharmaceutically acceptable dosage form through pharmaceutically acceptable auxiliary materials; the golden sargentgloryvine stem polysaccharide is prepared by the following method:

pulverizing and sieving the lygodium japonicum, taking powder, adding deionized water, carrying out hot reflux extraction, filtering an extracting solution, carrying out reduced pressure concentration, centrifuging, collecting supernatant, adding 3 times of volume of absolute ethyl alcohol, uniformly stirring, standing for alcohol precipitation, collecting precipitate, redissolving with deionized water, centrifuging, collecting supernatant, adding 3 times of volume of absolute ethyl alcohol, uniformly stirring, standing for alcohol precipitation, collecting precipitate, washing, redissolving with deionized water, deproteinizing by a Sevag method, and freeze-drying.

5. The pharmaceutical formulation of claim 4, wherein: the auxiliary material is solid, liquid or semisolid auxiliary material.

6. The pharmaceutical formulation of claim 4, wherein: the dosage forms comprise tablets, capsules and injections.

Technical Field

The invention belongs to the field of medicines, relates to development and application of a metabolism check point medicine, and particularly relates to a metabolism check point FBP1 agonist and application thereof in the aspect of tumor resistance.

Background

Glycolysis and oxidative phosphorylation are the major pathways for energy metabolism by cells. For normal cells, glucose is completely oxidized when oxygen is sufficient to produce 32 molecules of Adenosine Triphosphate (ATP); 2 molecules of ATP are produced only by glycolysis when oxygen supply is insufficient. Thus, normal cells use glycolysis, a low efficiency energy-producing pathway, only in the absence of oxygen. Tumor cells, even when they are sufficiently oxygenated, are still supplied with energy by glycolysis, which is a difference in sugar metabolism between tumor cells and normal cells, and is called the "Warburg (Warburg) effect", and since then, tumors are also considered to be a "metabolic disease". Based on this, the goal of tumor treatment by using drugs to regulate and control the carbohydrate metabolism of tumor cells has become a research hotspot in the fields of tumor cell biology and metabolism (carbohydrate metabolism regulation and tumor treatment of tumor cells, journal of biomedical engineering, 2019).

Since tumors are also a "metabolic disease," regulation of various metabolic functions of tumor cells has become an important research direction for tumor therapy. Recently, the research on the metabolism of tumor cells is very active, and the activation, differentiation, migration and the like of the tumor cells have key metabolic pathways and corresponding check points, so that the 'metabolic check points' refer to some important enzymes or receptors in the metabolic pathways, and the activity of the 'metabolic check points' directly influences the functions of the tumor cells and can regulate the proliferation, migration and invasion of the tumor cells. Then, by regulating the "metabolic checkpoint" of tumor cells to change their metabolic state, reduce the proliferation, migration and invasion activities of tumor cells, or reduce the tolerance of tumor cells to chemotherapeutic drugs, or reduce their immune escape capacity to immune cells, it is likely to be the next new key breakthrough point for improving the efficacy of cancer.

Fructose-1,6-bisphosphatase 1 (FBP 1), one of the key enzymes of gluconeogenesis, exerts an inhibitory effect during sugar degradation by catalyzing the hydrolysis of fructose-1, 6-diphosphate (upregulation of the inhibitory effect of FBP1 expression on gastric cancer cell growth, journal of gastroenterology and hepatology, 2018). FBP1 is low-expressed in various tumor cells, and is related to the growth and proliferation of tumor cells, and the growth and proliferation of tumor cells can be inhibited by activating FBP 1. Zhangieyi et al have studied the effect and mechanism of up-regulating FBP1 expression on gastric cancer cell proliferation and apoptosis, and found that FBP1 has low expression in gastric cancer cells, and up-regulating expression can inhibit cell proliferation and induce apoptosis, and the action mechanism may be related to up-regulating clear caspase-3 protein and down-regulating Cyclin D1 and c-Myc protein expression (up-regulating FBP1 expression has inhibition effect on gastric cancer cell growth, J gastroenterology and hepatology, 2018). The Hongzhengdong et al discuss the expression of FBP1 in human Renal Clear Cell Carcinoma (RCCC) and tissues beside the RCC, and the role and clinical significance in the occurrence, development and prognosis of renal cancer, and as a result, the FBP1 and the protein thereof are found to be low expressed in the human renal cancer tissue, are related to the occurrence and development of renal cancer, and may become one of candidate markers for renal cancer prognosis (the expression change and significance of FBP1 in the renal clear cell carcinoma tissue, Chongqing medicine, 2018). Liu Jia et al have discussed the expression level of FBP1 gene in colorectal adenocarcinoma and the relation between the expression level and clinical pathological features, and as a result, found that the low expression of FBP1 gene and protein may play an important role in the occurrence and development of colorectal adenocarcinoma, and is a potential therapeutic target (immunohistochemistry and qPCR technology for detecting the expression and clinical significance of FBP1 in colorectal adenocarcinoma, reported by Chuanbei medical college, 2017). It was found that FBP1 was low expressed in lung cancer tissues and cells, and that restoration of FBP1 expression not only inhibited glucose uptake and lactate production, but also inhibited lung cancer cell proliferation and invasion under hypoxic conditions in vitro, and inhibited lung cancer growth in vivo (Down-regulation of FBP1 by ZEB1-mediated expression controls to growth and invasion in lung cancer cells [ J ]. Molecular and Cellular Biochemistry, 2016).

Unfortunately, there are very few reports of FBP1 activators as the metabolic checkpoint and the studies are clearly deficient.

The traditional Chinese medicine LygodiiHerba is dry aerial parts of lygodium japonicum (Thunb.) Sw., lygodium japonicum L.microphyllum (Cav.) R.Br. or lygodium japonicum L.flexuosum (L.) Sw. of lygodium japonicum of the family lygodium japonicum, has the functions of clearing heat and removing toxicity, and inducing diuresis for treating stranguria, and contains various chemical components such as polysaccharide, flavone, terpenoid and the like.

At present, no report on the activity of the cissampelos aurantiaca polysaccharide for a metabolism check point FBP1 activator and an anti-tumor effect exists.

Disclosure of Invention

The invention aims to overcome the defects of the prior art and provides a metabolic checkpoint FBP1 agonist and application thereof in resisting tumors, particularly lung adenocarcinoma.

The above purpose of the invention is realized by the following technical scheme:

a FBP1 agonist as a metabolism check point is a polysaccharide of lygodium japonicum thunb, and is prepared by the following steps:

pulverizing and sieving the lygodium japonicum, taking powder, adding deionized water, carrying out hot reflux extraction, filtering an extracting solution, carrying out reduced pressure concentration, centrifuging, collecting supernatant, adding 3 times of volume of absolute ethyl alcohol, uniformly stirring, standing for alcohol precipitation, collecting precipitate, redissolving with deionized water, centrifuging, collecting supernatant, adding 3 times of volume of absolute ethyl alcohol, uniformly stirring, standing for alcohol precipitation, collecting precipitate, washing, redissolving with deionized water, deproteinizing by a Sevag method, and freeze-drying.

The medicinal application of the sargentgloryvine stem polysaccharide in preparing antitumor medicaments.

Further, the tumor is lung adenocarcinoma, liver cancer or breast cancer.

A pharmaceutical preparation for treating lung adenocarcinoma comprises a polysaccharide of radix Stephaniae Sinicae as active ingredient, and pharmaceutically acceptable adjuvants to make into pharmaceutically acceptable dosage forms; the golden sargentgloryvine stem polysaccharide is prepared by the following method:

pulverizing and sieving the lygodium japonicum, taking powder, adding deionized water, carrying out hot reflux extraction, filtering an extracting solution, carrying out reduced pressure concentration, centrifuging, collecting supernatant, adding 3 times of volume of absolute ethyl alcohol, uniformly stirring, standing for alcohol precipitation, collecting precipitate, redissolving with deionized water, centrifuging, collecting supernatant, adding 3 times of volume of absolute ethyl alcohol, uniformly stirring, standing for alcohol precipitation, collecting precipitate, washing, redissolving with deionized water, deproteinizing by a Sevag method, and freeze-drying.

Further, the auxiliary material is solid, liquid or semisolid auxiliary material.

Further, the dosage forms comprise tablets, capsules and injections.

Has the advantages that:

the invention provides an FBP1 activator as a metabolic checkpoint, wherein the activator is a natural plant polysaccharide, and CCK-8 tests prove that the activator can obviously inhibit the proliferation of human lung adenocarcinoma, liver cancer and breast cancer cells and presents obvious dose dependence; western blotting experiments prove that the sargentgloryvine stem polysaccharide can effectively activate the expression of FBP1 protein in the tumor cells, which is probably an action mechanism of the sargentgloryvine stem polysaccharide for resisting human lung adenocarcinoma, liver cancer and breast cancer.

Drawings

FIG. 1 shows the proliferation inhibition rate of Cissampelos segetum polysaccharide on human lung adenocarcinoma A549 cells at different concentrations;

FIG. 2 is a graph showing the effect of different concentrations of sargentgloryvine stem polysaccharide on the expression of FBP1 protein in human lung adenocarcinoma A549 cells;

FIG. 3 shows the proliferation inhibition rate of Cissampelos segetum polysaccharide in different concentrations on human liver cancer HepG2 cells;

FIG. 4 shows the effect of different concentrations of sargentgloryvine stem polysaccharide on the expression of FBP1 protein in human liver cancer HepG2 cells;

FIG. 5 shows the proliferation inhibition rate of Cissampelos segetum polysaccharide in different concentrations on human breast cancer MDA-MB-231 cells;

FIG. 6 shows the effect of different concentrations of sargentgloryvine stem polysaccharide on the expression of FBP1 protein in human breast cancer MDA-MB-231 cells.

Detailed Description

The following detailed description of the present invention is provided in connection with the accompanying drawings and examples, but not intended to limit the scope of the invention.