WO2023040311A1

WO2023040311A1 - Application of cationic polymer in preparation of drug for removing intestinal microbial toxins and treating tumors

Info

Publication number: WO2023040311A1
Application number: PCT/CN2022/093060
Authority: WO
Inventors: 韩源平
Original assignee: 成都施桂行医药科技有限责任公司
Priority date: 2021-09-16
Filing date: 2022-05-16
Publication date: 2023-03-23
Also published as: CN114099540A; CN114099540B

Abstract

An application of a cationic polymer in the preparation of a drug for treating or preventing tumors. Oral administration of the cationic polymer can alleviate, treat and prevent various cancers, and alleviate complications thereof. Oral administration of the cationic polymer can effectively remove or eliminate metabolites produced by intestinal microorganisms, remove endotoxins produced by bacteria, reduce the entry of endotoxins into the blood, reduce the entry of bacterial CpG-DNA into the blood, and reduce the entry of bacterial flagellin into the blood. Oral administration of the cationic polymer can reduce system inflammation and tumor environment inflammation of the body, can improve the tumor immune environment, activate cell autophagy to inhibit tumor growth and metastasis, and reduce tissue fibrosis, including liver cirrhosis and pancreatic fibrosis. The cationic polymer can be used in combination with other anti-tumor therapies, including radiotherapy, chemotherapy and immunotherapy, to improve efficacy and tolerance.

Description

Application of cationic polymer in the preparation of drugs for removing intestinal microbial toxins and treating tumors

technical field

The invention relates to the field of medicine, in particular to the application of oral cationic polymers in the preparation of drugs for removing various toxins in the intestinal tract to treat or prevent tumors.

Background technique

Incidence of pancreatic cancer. Worldwide, the incidence of cancer increases by 19 million people every year, and pancreatic cancer increases by 460,000 people every year. In 2015 alone, there were approximately 90,100 new cases of pancreatic cancer and 79,400 deaths from pancreatic cancer in my country. The 5-year survival rate of patients with pancreatic cancer worldwide is approximately 6%. Pancreatic cancer ranks as the 14th most common cancer and the 7th cause of cancer death in the world. The exact cause of pancreatic cancer is unknown. Risk factors include diabetes, chronic pancreatitis, and gastric ulcers. There are no obvious symptoms in the early stage of pancreatic cancer, but weight loss and abdominal pain are common symptoms in the later stage. Pancreatic cancer is often diagnosed at an advanced stage. Only 20% of pancreatic cancer patients can be surgically removed when they develop. Among patients who were able to undergo successful surgical resection, the 5-year survival rate was 27%, compared with 6-11 months and 2-6 months for patients with locally advanced or metastatic disease, respectively.

Factors that cause pancreatic cancer. Although the cause of pancreatic cancer is not completely clear, many factors can promote the occurrence and development of pancreatic cancer. Among the various causes of pancreatic cancer, advanced age (old age) is the greatest risk factor for pancreatic cancer. Most cancers occur in old age, especially pancreatic cancer. Pancreatic cancer is typically a disease of the elderly. It is extremely rare for patients to be diagnosed before the age of 30, with 90% of newly diagnosed patients being over the age of 55, with the majority in their 70-80s. Why pancreatic cancer occurs more in the elderly, the mechanism is still unclear. Recently, through animal models, we proposed a new mechanism for the late development of pancreatic cancer. We believe that the special weak extracellular matrix (ECM) of the pancreas can confine tumor gene-transformed cancer cells to a long-term dormant state.

Sex differences in pancreatic cancer. Worldwide, the incidence of pancreatic cancer is higher in men than in women (5.1 for men and 4.0 for women). This gap appears to be larger in high development index countries. Despite significant sex differences in pancreatic cancer, a systematic review of 15 studies concluded that female reproductive factors are not associated with pancreatic cancer.

Family history and genetic predisposition to pancreatic cancer. Pancreatic cancer is considered to be familial if two or more first-degree relatives have been previously diagnosed with pancreatic cancer. Pancreatic cancer due to genetic factors accounts for 5%-10% of new cases. Patients with familial risk factors were nine times more likely to develop pancreatic cancer than those without a family history.

Diabetes and pancreatic cancer. Diabetes is a recognized risk factor for pancreatic cancer. The study found that type 1 diabetes can double the risk of pancreatic cancer. Patients with type 2 diabetes had a similar increased risk of pancreatic cancer.

Relationship between lifestyle and pancreatic cancer. Smoking is considered the most important modifiable risk factor for pancreatic cancer. Compared with never smokers, current smokers had a 74% increased risk of developing pancreatic cancer. Excessive alcohol consumption is also a major cause of chronic pancreatitis, a known risk factor for pancreatic cancer, so drinking alcohol in this setting is a risk factor for pancreatic cancer. The risk increases significantly when the patient drinks more than 30g of alcohol per day. Among people who drink heavily, there is a 15 percent increased risk of pancreatic cancer. Numerous studies have found that consumption of red and processed meat is associated with the development of pancreatic cancer. Excessive consumption of red and processed meat has been shown to have the potential to cause DNA damage and the formation of carcinogens, such as N-nitroso compounds. Other dietary factors with limited suggestive evidence for pancreatic cancer etiology include foods and beverages that contain fructose, or foods that contain saturated fatty acids. An increased risk of pancreatic cancer has been observed in patients infected with Helicobacter pylori (H. pylori) or hepatitis C virus.

Chronic pancreatitis and pancreatic cancer. This is an important risk factor for pancreatic cancer. Chronic pancreatitis is a progressive inflammatory state of the pancreas that results in pancreatic fibrosis and loss of acinar and islet cells. About 5% of patients will develop pancreatic cancer during their lifetime. These patients had a significantly 13-fold increased risk of pancreatic cancer compared with the general population or controls.

Obesity and Pancreatic Cancer: The prevalence of obesity is rising globally, with an estimated 2 billion adults and 300 million children and adolescents globally classified as overweight or obese. Association between elevated body mass index (BMI) and pancreatic cancer. Increased risk of pancreatic cancer for every 5 BMI units. Rising rates of obesity are likely to be a major factor in the rise in pancreatic cancer rates in developed countries.

Ductal adenocarcinoma: This is the most common pancreatic cancer (Pancreatic ductal adenocarcinoma, PDAC), which is an exocrine malignancy. Other pancreatic cancers include neuroendocrine tumors, which start in neuroendocrine cells. Pancreatic ductal adenocarcinoma (PDAC) accounts for 90% of all pancreatic cancers. About 60%-70% of pancreatic ductal adenocarcinomas occur in the head of the pancreas, and the rest in the body (15%) and tail (15%) of the pancreas. At the time of diagnosis, most pancreatic adenocarcinomas have spread beyond the pancreas, including lymph node metastasis. Ductal adenocarcinoma accounts for the majority of pancreatic cancers, which are mainly composed of duct-like glands with varying degrees of differentiation, accompanied by abundant fibrous stroma (fibrosis). Well-differentiated ductal adenocarcinoma mainly consists of well-differentiated duct-like structures lined with tall columnar epithelium, some with myxoid epithelium and some with abundant eosinophilic cytoplasm. Cancers vary in size, and small adenocarcinomas can be buried in the pancreatic parenchyma, but the surrounding pancreatic tissue is often hardened, and sometimes the pancreas can be deformed. Pancreatic cancer is sometimes difficult to distinguish from chronic pancreatitis, so that chronic pancreatitis is mistaken for pancreatic cancer and undergoes radical surgery. Cancer of the head of the pancreas often invades the duodenal wall. Most cancers are located in the head of the pancreas, accounting for 70% to 80%. Cancer of the head of the pancreas may have common bile duct obstruction in the early stage, and gradually aggravating jaundice clinically. Cancer of the body and tail of the pancreas is often clinically without jaundice, but often produces ascites due to invasion of the portal vein by cancer tissue, splenomegaly due to compression of the splenic vein, and deep pain due to invasion of the celiac plexus. Sometimes the levels of trypsin, leucine aminopeptidase, and amylase in the patient's serum are all increased, but because the results are not constant, they are not often used in clinical diagnosis.

Biological pathogenesis of pancreatic cancer. The biogenesis of pancreatic cancer evolves slowly over a series of decades. Normal pancreatic exocrine cells, including pancreatic ductal epithelial cells, progress through intermediate stages of specific precursor lesions to eventually form invasive malignancies. The three most typical precursors of this malignancy are pancreatic intraepithelial neoplasia (PanIN), intraductal papillary mucinous neoplasms (IPMN), and mucinous cystic neoplasms (mucinous cystic neoplasms). ,MCN). These distinct stages have specific clinical features as well as pathological and molecular features. Pancreatic intraepithelial neoplasia (PanIN) is a noninvasive microscopic lesion that occurs in small (usually less than 0.5 cm) pancreatic ducts. PanIN may be the result of localized pancreatitis, and the resulting repeated epithelial damage and repair may further lead to the occurrence of malignancy. PanIN can be further graded into grades 1–3, reflecting the degree of progressive tumor morphological changes. Pancreatic intraepithelial neoplasia (PanIN) is an abnormal growth of the pancreatic epithelium that results in "dysplasia" of the pancreatic ductal epithelium that can lead to loss of the normal tissue outcome of the pancreas.

Somatic genetic mutations in pancreatic cancer. The PanIN lesion stage was often accompanied by the presence of the KRAS oncogene and exhibited shortened telomeres, suggesting that these are early alterations in an aggressive malignancy pathway. Mutations continue to accumulate with age, and mutations in tumor suppressor genes such as P16, CDNK27, P53, and SMAD4 also appear later in PanIN. The KRAS mutation rate also increased further with increasing PanIN grades. Abnormalities in Notch signaling and the sonic-hedgehog pathway have also been implicated in pancreatic carcinogenesis. About 80 percent of these mutations are acquired sporadically. The accumulation of these multiple mutations further promotes the transformation of cells, such as epithelial to mesenchymal transition (EMT), further de-differentiation of tumor cells, detachment from the pancreatic duct, and transfer to other cells. Tissues proliferate further. The slow accumulation of tumor gene mutations is also an important reason for the late onset of pancreatic cancer in old age.

Diagnosis of pancreatic cancer. The early symptoms of pancreatic cancer are not typical, often manifested as epigastric discomfort, low back pain, indigestion or diarrhea. Patients have loss of appetite and weight loss, and most of them are in the middle and late stages when symptoms appear.

(1) Laboratory examination: CA19-9 is currently the most commonly used diagnostic marker for pancreatic cancer, and has the following clinical features: Serum CA19-9>37U/ml is used as a positive indicator, and the sensitivity and specificity of diagnosing pancreatic cancer reach 78.2% respectively. % and 82.8%. About 10% of pancreatic cancer patients are negative for Lewis antigen and CA19-9 is not elevated. At this time, other tumor markers such as CA125 and/or carcinoembryonic antigen (CEA) should be combined to assist in the diagnosis. For patients with elevated CA19-9, pancreatic cancer should be highly suspected after excluding factors such as biliary obstruction or biliary system infection. Changes in blood sugar have also been linked to pancreatic cancer onset or progression.

(2) Imaging examination: Enhanced three-dimensional dynamic CT thin-layer scan is currently the most commonly used method for diagnosing pancreatic cancer. It can clearly display the size, location, density and blood supply of the tumor, and judge the tumor and blood vessels accordingly (CT should be used if necessary). Angiography) and the adjacency relationship of adjacent organs guide the resectability of the tumor before surgery and the evaluation of the effect of neoadjuvant chemotherapy. In addition to showing the anatomical features of pancreatic tumors, MRI can also clearly show the presence or absence of metastatic lesions in the parapancreatic lymph nodes and liver; and it is superior to CT in distinguishing from edematous or chronic mass pancreatitis. The combined application of magnetic resonance cholangiopancreatography and MRI thin-slice dynamic enhancement is helpful to clarify pancreatic cystic and solid lesions. Positron emission tomography (PET)-CT examination images can reveal the metabolic activity and metabolic burden of tumors, and have obvious advantages in finding extrapancreatic metastases and evaluating systemic tumor burden. Endoscopic ultrasonography (EUS) combined with ultrasound imaging on the basis of endoscopic technology has improved the sensitivity and specificity of pancreatic cancer diagnosis.

(3) Pathological examination: Histopathological and/or cytological examination is the "gold standard" for the diagnosis of pancreatic cancer. Except for patients who are scheduled to undergo surgical resection, the rest of the patients should strive to clarify the pathological diagnosis before formulating a treatment plan. Current methods for obtaining histopathological or cytological specimens include: EUS or CT-guided needle biopsy; exfoliated cytology of ascitic fluid; and exploratory biopsy under laparoscopy or laparotomy.

Pancreatic cancer treatment. The preferred method is surgery, followed by radiotherapy and chemotherapy. As new explorations, immunotherapy and gene therapy have gradually begun to be used in the treatment of pancreatic cancer.

Surgical treatment of pancreatic cancer: In general, only 20% of patients with PDAC receive surgical treatment. There are generally four surgical options. During the treatment of the disease, patients should also prescribe the right medicine and assist with chemotherapy. The multi-center incidence of pancreatic cancer has attracted more and more attention. In addition to the main tumor in the head of the pancreas, multiple small tumors can also be found in other parts of the entire pancreas. Resection provides an important rationale.

Chemotherapy for pancreatic cancer: Chemotherapy strategies mainly include postoperative adjuvant chemotherapy, neoadjuvant chemotherapy, and palliative chemotherapy for patients with locally advanced unresectable or distant metastasis. Although chemotherapy is not the preferred treatment for pancreatic cancer, it can be used as adjuvant chemotherapy after pancreatic cancer surgery, mainly gemcitabine, combined with other drugs, can prolong survival. Chemotherapy generally has relatively strong side effects. Taking it together with ginsenoside Rh2 can relieve the discomfort caused by side effects and even promote the effect of chemotherapy drugs. The recommended adjuvant chemotherapy regimen is gemcitabine or fluorouracil drugs [including capecitabine, S-1, 5-fluorouracil (5-FU) combined with leucovorin] as monotherapy. Gemcitabine is a pyrimidine nucleotide analog, which belongs to the anti-metabolism anticancer drug dFdCTP and dCTP compete with dCTP to be incorporated into the DNA chain to extend the DNA chain. DNA polymerase cannot remove the incorporated dFdCTP, so that the extended DNA chain cannot be repaired. This inhibits DNA synthesis and eventually leads to apoptosis. It has the characteristics of broad anti-cancer spectrum, unique mechanism of action, low toxicity, no cross-resistance with other chemotherapeutic drugs, and no superposition of toxicity. The FOLFIRINOX four-drug combination regimen prolongs pancreatic cancer, including oxaliplatin, irinotecan, fluorouracil, and leucovorin. IRIN: irinotecan irinotecan, OX: Oxaliplatin (oxaliplatin). Pancreatic cancer has a high degree of malignancy, from discovery to death very quickly, so the effect of chemotherapy on prolonging survival is very limited. Chemical drugs can directly inhibit or kill cancer cells, but many chemotherapy drugs lack selectivity and cause damage to the body, causing toxic side effects such as the hematopoietic system, immune system, digestive system, and nervous system.

Immunotherapy of pancreatic cancer: In recent years, although immunotherapy has made important progress in the treatment of a variety of solid tumors, this type of new anti-cancer strategy is helpless against the "king of cancer" pancreatic cancer, whether it is PD-1/ Neither PD-L1 antibody nor CTLA-4 antibody monotherapy, nor the combination of the two drugs, has been significantly successful in improving survival in patients with pancreatic cancer. The reasons for the failure of immunotherapy for pancreatic cancer are being explored. This may be related to gut flora.

The relationship between pancreatic cancer and the gut microbiome. Microbiota, including bacteria, viruses, fungi, and archaea. The large intestine of an adult contains 1 kilogram of microorganisms, which is a collection of trillions of microorganisms. The microbes in the gut have the following physiological functions: (A) defend against foreign organisms, because of their huge numbers, gut microbes can destroy small amounts of microbes from dietary sources; (B) help digest food residues that have not been broken down in the small intestine, Produce short chain fatty acids (short chain fatty acids); (C) produce B vitamins, these B series vitamins are key coenzymes for human metabolism, and are extremely important for this metabolic balance; (D) promote the body's immune system. The intestine is an important innate immune barrier of the body; at the same time, about 40% of lymphocytes gather in the intestine. Gut microbes are extremely important to the body's immune system. The microstate of intestinal flora is determined by many factors. First of all, the intestinal flora of the fetus comes from the mother; then the dietary components and the body's innate immune system also shape the enterotype. In recent years, our work has found that vitamin D also shapes gut microbiota through the vitamin D receptor (VDR) of the small intestine.

The imbalance of intestinal microbiosis (dysbiosis) can lead to a variety of metabolic diseases. Gut microbes, as well as microbes present in the pancreas, can contribute to the development of pancreatic cancer. The pancreas was once considered a sterile organ, and recent research has shown that there is a microbiome in the pancreas, but its role is still uncertain. Geller and colleagues examined 113 human PDAC samples obtained during surgery and 20 normal pancreas samples from organ donors by performing quantitative polymerase chain reaction (qPCR) on bacterial 16S ribosomal DNA. They found that 76 percent of pancreatic tumor samples and 15 percent of normal pancreas contained bacteria. Further 16S rRNA gene sequencing was performed on 65 PDAC samples, and the results showed that the abundance of the bacteria γ-proteobacteria in these samples was high, which is a Gram-negative bacterium that can produce a large amount of endotoxin (endotoxin). Can activate inflammatory response and promote tumor growth. Similarly, an independent 16S rRNA gene sequencing study of 12 PDAC samples by Pushalkar et al. also demonstrated the presence of microorganisms but identified the ubiquitous presence of Proteobacteria, Bacteroidetes, and Firmicutes in all samples; moreover, they found significant differences in the microbial composition between human pancreatic tumor samples and normal human pancreases. However, Thomas and colleagues found that the intestinal flora of mice with Kras G2D/PTENlox/+ genetic background can promote the progression of pancreatic cancer; original pancreatic cancer, suggesting that the bacteria in the pancreatic cancer authors may not be important, but may be a remote role in the gut. Removing bacteria from the pancreas restores sensitivity to checkpoint blockade immunotherapy. Gut microbes can translocate to the pancreas and induce an immunosuppressive tumor microenvironment that contributes to the progression of PDAC. Therefore, using antibiotics to reduce microorganisms in tumors can improve/enhance tumor immunotherapy, such as improving the efficacy of PD-1 antibodies. Similarly, the use of antibiotics to eliminate microbes in tumors also reduces myeloid-derived suppressor cells (MDSCs), reduces macrophages from immunosuppressive M2 cells, and tumor-associated macrophages (TAMs), promoting their migration to M1-like Differentiation, while increasing the tumor suppressor function of Th1-CD4+ T cells and cytotoxic CD8+ T cells. Antibiotic treatment can induce upregulation of PD-1 expression in CD4+ and CD8+ T cells, and when PD-1 blockade is used in combination with antibiotics, it can enhance the synergistic effect, thereby significantly reducing tumor size.

Failure of chemotherapy/immunotherapy in pancreatic cancer in relation to gut microbiota. Geller and colleagues describe the ability of bacteria belonging to the Gamma-proteobacteria class to metabolize the chemotherapy drug gemcitabine (2′,2′-difluorodeoxycytidine) to its inactive form 2′,2′-difluorocytidine deoxyuridine. Using a mouse model of pancreatic cancer, they demonstrated that the presence of Gamma-proteobacteria within tumors is responsible for the development of resistance to gemcitabine. The use of antibiotics eliminated this effect. Since gemcitabine is already used in advanced pancreatic cancer, they speculated that the presence of this bacterium might lead to resistance to this drug. In fact, they detected Gamma-proteobacteria in tissue specimens from 76 percent (86/113) of human pancreatic ductal adenocarcinoma (PDAC) patients, suggesting that tumor samples contained bacteria that might modulate tumor sensitivity to gemcitabine. This study highlights the importance of the microbiome outside the gut, particularly intratumoral bacteria, in altering the natural history of this cancer.

Correlation between pancreatic carcinogenesis and intestinal bacterial endotoxin. The innate immune system consists of epidermal cell shields and a variety of non-specific immune cells. At the molecular level, the membrane receptor family of Toll-like receptors (TLRs) recognize various components of a variety of bacterial pathogens. Endotoxin (lipopolysaccharide, LPS) is specifically recognized by TLR4, thereby triggering gene expression, which can control the innate immune response and further guide the development of antigen-specific acquired immunity on the one hand; on the other hand, it can lead to non-specific Inflammation. LPS can act not only on immune cells, but also certain types of epithelial cells, including cancer cells, and promote their transformed phenotypes. Specifically, LPS can activate NF-kappaB signaling in pancreatic cancer cells, linking inflammation to cancer progression. In vitro experiments have shown that human pancreatic cancer cells Panc-1 and AsPC-1 activate the TLR4/NF-kappaB signaling pathway under LPS stimulation, leading to increased invasion ability. Studies have found that the expression of TLR4 in pancreatic cancer cells and PDAC tissues increases. After LPS treatment, the migratory ability of pancreatic cancer cells increased, while the protein level of the tumor suppressor PTEN decreased. Transgenic mice with high expression of KRas gene in the pancreas (LSL-Kras/G12D) can mimic most of the phenotypes of human pancreatic cancer. Studies have found that as the disease progresses, blood lipids decrease and lactic acid and taurine increase. A study on the flora ecology of the duodenum of patients with pancreatic cancer found that not only the plasma endotoxin, IL-6, and CRP (C-creatiine protein) levels increased, but also the intestinal mucosa was damaged, and Helicobacter pylori (H. pylori ) also increased, further indicating that intestinal damage and flora disorder can promote the development of pancreatic cancer.

Chronic pancreatitis and hyperbile acidemia (Hyperbileacidemia) are common in the later stages of pancreatic cancer. Various risk factors for pancreatic cancer, including alcohol consumption, smoking, high-fat diet, diabetes, etc., can increase the risk of pancreatitis and hyperbiliary acidemia. In addition, the proliferation of pancreatic cancer will compress the biliary tract, resulting in obstruction, resulting in bile fluid backflow and hyperbiliary acidemia. High bile acids are closely related to various cancers of the digestive tract. At the cellular level, bile acids can promote the expression of cyclooxygenase-2 expression, which in turn promotes inflammation in the pancreas.

Therefore, a high molecular weight cationic polymer and its preparation method are provided, as well as a drug for removing various acidic products of intestinal microorganisms, including endotoxins, to treat, alleviate, and prevent specific diseases or a combination of multiple diseases The application in it has important practical significance.

Contents of the invention

In view of this, the present invention provides a high-molecular cationic polymer and its preparation method, as well as its application in medicine for treating, alleviating and preventing a specific disease or a combination of multiple diseases.

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 the cationic polymer in the preparation of preparations or medicines for eliminating or neutralizing the pathogenic factors produced by intestinal microorganisms through oral administration.

In some specific embodiments of the present invention, the pathogenic factors include but are not limited to: endotoxin (lipopolysaccharide), negatively charged short-chain fatty acids (including acetic acid, propionic acid, butyric acid), hydrogen sulfide, One or more of negatively charged microbial derivatives and decomposition products, bile acids, bacterial DNA fragments (CpG-DNA), nucleotides, and flagellin produced by intestinal bacteria.

Based on the above research, the present invention also provides the application of orally administered cationic polymers in the preparation of preparations or medicines for eliminating endotoxins and other derivatives produced by intestinal microorganisms.

The present invention also provides the special application of high molecular cationic polymer in the preparation of preparations or medicines for preventing, alleviating, improving and/or treating diseases caused by endotoxin produced by intestinal microorganisms.

In some specific embodiments of the present invention, the disease includes systemic inflammation of the tumor body, local inflammation of the tumor, tissue fibrosis, and tumor immune tolerance;

The diseases near the systemic inflammation include one or more of diabetes, fatty liver and tumor.

The preparation or drug reduces the AKT/mTOR pathway to inhibit tumor growth, activates the autophagy-lysosome pathway, removes degenerated/damaged organelles in cells, decomposes YAP, and inhibits the growth of tumor cells by reducing the endotoxin content in the blood. Growth, inhibition of tumor cell migration, promotion of autophagy - increased lysosomal flux, reduction of oncogene expression and/or reduction of cellular stress state.

In some embodiments of the invention, the cationic polymers include:

(1) Multiple positive charges; and

(2) The skeleton is an organic covalent bond polymer polymer;

Preferably, the high-molecular cationic polymer also includes a hydrophobic structure.

In some embodiments of the present invention, the high-molecular cationic polymer includes a polyamine polymer; the polyamine polymer includes repeating units derived from the polymerization of amine monomers and crosslinking monomers;

Preferably, the polyamine polymer is an organic polymer containing tertiary amine or quaternary amine;

Most preferably, the amine polymer includes Choestyramine, Colestipol, and Colesevelam, which are used to remove various toxins produced by intestinal microorganisms;

The molecular weight of the high-molecular cationic polymer is greater than 4000 Da, preferably, the molecular weight of the high-molecular cationic polymer is 1×10 ⁶ ~10×10 ⁶ Da;

The polymeric properties and the water insolubility of the polymer are structurally critical for this application, with safety and tolerance characteristics.

The endotoxin includes lipopolysaccharides (lipopolysaccharides, LPS) of bacterial cell walls;

The endotoxin is mainly present in the outer membrane of Gram-negative bacteria;

The cancer includes one or more of pancreatic cancer, liver cancer, rectal cancer, gastric cancer, esophageal cancer, bladder cancer, kidney cancer, lung cancer, breast cancer, head and neck cancer, and skin cancer;

The prevention, alleviation, improvement and/or treatment include alleviating or inhibiting one or more of the growth, transformation, deterioration and metastasis of tumor cells;

Preferably, the prevention, mitigation, improvement and/or treatment include any of the following items or any combination thereof:

(1) Relieve the progression of pancreatic cancer and inhibit the malignant transformation of pancreatic intraepithelial neoplasia (PanIN) to pancreatic ductal adenocarcinoma (PDAC); and/or

(2) Inhibit local invasion and malignant systemic metastasis of pancreatic cancer; and/or

(3) Inhibition/remission of complications of pancreatic cancer, such as jaundice; and/or

(4) Inhibit/relieve pancreatic fibrosis and liver fibrosis, inhibit liver cirrhosis in cancer patients; and/or

(5) Reduce the endotoxin content in the blood of cancer patients, reduce endotoxemia, reduce the increase of transaminase in the blood, and improve liver function; and/or

(6) Reduce the various bile acid components in the blood of cancer patients, reduce the bilirubin content in the blood, and relieve the symptoms of jaundice; and/or

(7) reduce cancer systemic inflammatory storm, reduce systemic inflammation, reduce local inflammation in lesion tissue; and/or

(8) Promote the intestinal flora balance (eubiosis) of cancer patients;

Based on the above research, the present invention also provides a drug or a pharmaceutical composition, including a high-molecular cationic polymer and a pharmaceutically acceptable adjuvant; the pharmaceutical composition also includes other arbitrary active ingredients and a pharmaceutically acceptable adjuvant.

The present invention relates to pharmaceutical and medical applications of cationic polymers. Through oral administration of the cationic high molecular polymer, the present invention can remove various acidic derivatives produced by intestinal microorganisms, so as to alleviate, treat, prevent various tumors and cancers, and relieve the complications of these tumors. The core of the present invention is a polymer compound that is not degraded or absorbed by the human body when taken orally, including polymers rich in cations, such as polystyrene quaternary ammonium salt (Cholestyramine), cholestyramine Colestipol, Colesevelam and other analogues. Through its unique molecular affinity, the application of the invention can effectively remove or eliminate metabolites produced by intestinal microorganisms, including various toxins produced by various bacteria, such as endotoxin, bacterial CpG-DNA, and bacterial flagellin. This application can effectively reduce the entry of endotoxin into the blood, reduce the entry of CpG-DNA into the blood, reduce the entry of bacterial flagellin, reduce the entry of bile acid into the blood, and reduce the entry of bilirubin into the blood. This application can effectively reduce the systemic inflammation of the body and the inflammation of the tumor environment, and can improve the tumor immune environment. This application can also activate autophagy to inhibit tumor growth and metastasis; reduce tissue fibrosis including liver cirrhosis and pancreatic fibrosis; and promote damage repair. The high-molecular cationic polymer that is not absorbed by the human body can also be used in combination with other anti-tumor therapies, including radiotherapy, chemotherapy, and immunotherapy, so as to improve the curative effect and tolerance.

Description of drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings that are required in the description of the embodiments or the prior art.

Figure 1 shows the inhibition of polycationic polymer (cholestyramine) on mouse pancreatic cancer; wherein, Figure 1A shows that KPC mice given oral cholestyramine can inhibit the growth of pancreatic cancer; 6 weeks old WT (C57BL/6J ) mice and 6-week-old pancreatic cancer KC (PDX1-KrasG12D) mice were divided into three groups: (1) WT mice were given maintenance feed (blank control group, n=10); (2) pancreatic cancer KC (PDX1-KrasG12D) mice (3) pancreatic cancer KC (KrasG12D) mice were given maintenance feed containing 3% cholestyramine (experimental group, n=6); continuous feeding 10 weeks; Figure 1 shows the statistics of pancreatic lesion status and pathological changes of mice in each group at the end of the experiment; as shown in Figure 1A, taking cholestyramine can effectively inhibit the growth of pancreatic cancer in the three time periods of "early-middle-late" State (area ratio of pancreatic cancer in the whole tissue of pancreatic tissue section); Figure 1B shows pancreatic cancer histology, and the growth of pancreatic cancer can be effectively inhibited by oral cholestene under the PDX-Kras genetic background;

Figure 2 shows that oral administration of polycationic polymers (cholestyramine) inhibits the rectal metastasis of pancreatic cancer in mice, and inhibits the progression of pancreatic cancer metastasis; 6-week-old WT (C57BL/6J) mice and 6-week-old pancreatic cancer KC (PDX1 -KrasG12D) mice were divided into three groups: (1) WT mice were given maintenance feed (blank control group, n=10); (2) pancreatic cancer KC (PDX1-KrasG12D) mice were given maintenance feed (control group, n=6); (3) pancreatic cancer KC (KrasG12D) mice were given maintenance feed containing 3% cholestyramine (experimental group, n=6); continued to feed for 10 weeks; Figure 2 shows pancreatic tumor metastasis, Under the PDX-Kras genetic background, tumor metastasis is mainly manifested in the case of tumor rectal metastasis; the pancreatic cancer metastasis shown in Figure 2 also includes the frequency of intraocular and other visceral metastasis; as shown in Figure 2, polycationic polymers can The cancer metastasis rate was reduced from 63% in the control group to 36%; as shown in Figure 2, the state of biliary stasis was also significantly improved;

Figure 3 shows that hyperbiliary acidemia/jaundice caused by biliary tract ligation can promote the development and deterioration of pancreatic cancer; biliary obstruction and jaundice promote the development of pancreatic cancer; patients with pancreatic cancer are often accompanied by complications such as biliary obstruction and jaundice; for To verify its causal relationship, we adopted bile duct ligation; 9-week-old pancreatic cancer KC (PDX1-KrasG12D) mice were divided into two groups: (1) mice were given a sham operation of common bile duct ligation (control group, n=12); 2) The mice were given common bile duct ligation, and the common bile duct obstruction of the mice was formed, and then they were sacrificed after two weeks of continuous feeding (n=15); Figure 3A is a schematic diagram of the experimental design; Figure 3B shows the histobiological state of mouse pancreatic lesions; Bile duct ligation promotes the growth and metastasis of pancreatic cancer;

Figure 4 shows that bacterial endotoxin (LPS) promotes the development of pancreatic cancer; endotoxin produced by intestinal bacteria is an important cause of systemic inflammation as well as local inflammation in the tumor environment; we determined whether endotoxin can aggravate PDX-Kras genetic background mice Pancreatic cancer; here, 9-week-old pancreatic cancer KC (PDX1-KrasG12D) mice were divided into two groups: (1) KC mice were given intraperitoneal injection of 100 microliters of normal saline (control group, n=12); (2 ) gave KC mice 4 mg/kg intraperitoneal injection of Escherichia coli LPS, once a week, and collected mice after four weeks; wherein, Figure 4A shows a schematic diagram of the experimental design; Figure 4B shows pancreatic tissue staining; HE staining shows that giving LPS can aggravate The growth of pancreatic cancer, and Masson Trichrome staining showed that LPS can aggravate the fibrosis of pancreatic cancer tissue; Figure 4C shows the immunochemical staining of pancreatic tissue; Ck19 indicates the proliferation of ductal carcinoma, K67 indicates the proliferation of tumor cells, and Collagen type-1 indicates Fibrosis of pancreatic tissue;

Figure 5 shows that the blood bile acid in pancreatic cancer mice is elevated, and oral administration of cholestene can reduce the serum total bile acid content; due to the growth of pancreatic tumors, bile duct compression and cholestasis are often caused, resulting in hyperbiliary acidemia and jaundice, the latter Can promote tumor transformation and malignancy; Figure 5A shows that the bile acid content in serum of pancreatic cancer mice is significantly higher than that of control WT; Figure 5B shows that administration of polystyrene quaternary ammonium salt can significantly improve serum total bile acid levels;

Figure 6 shows that taking cholestyramine can effectively reduce the endotoxin content in the blood of mice with pancreatic cancer; here, we measured the content of endotoxin in the serum of mice with primary pancreatic cancer; 6-week-old WT mice and Six-week-old pancreatic cancer KC (PDX1-KrasG12D) mice were divided into three groups: (1) WT mice were given maintenance diet (blank control group, n=10); (2) pancreatic cancer KC (PDX1-KrasG12D) mice were given Rats maintained diet (control group, n=6); (3) gave pancreatic cancer KC (KrasG12D) mice a maintenance diet containing 3% cholestyramine (experimental group, n=6); continued to feed for 10 weeks; Figure 6 shows that at the end of the experiment, the LPS content in the serum, taking polystyrene quaternary ammonium salt can effectively reduce the endotoxin content in the serum of pancreatic cancer mice;

Figure 7 shows that oral administration of cholestyramine can reduce pancreatic fibrosis and ductal hyperplasia in mice with pancreatic cancer; tissue fibrosis is an important reason for the deterioration of pancreatic cancer and an important basis for the tumor microenvironment; while continuous tissue damage and chronic inflammation are An important cause of tissue fibrosis; therefore, eliminating fibrosis and clearing the tumor environment is a strategy for tumor therapy; we speculate that clearing intestinal endotoxins may reduce persistent inflammation in cancer tissues, thereby eliminating fibrosis in tumor tissues and inhibiting tumors hyperplasia; for this reason, 6-week-old WT mice and 6-week-old KPC mice were divided into three groups: (1) WT mice were given maintenance feed (blank control group n=10); (2) pancreatic cancer KC mice were given Maintenance diet (control group n=30); (3) feeding pancreatic cancer KC (PDX1-KrasG12D) mice with maintenance diet containing 3% cholestyramine (experimental group n=28) for 10 weeks; wherein, Fig. 7A Fig. 7B shows the semi-quantitative scan data, showing that taking polystyrene quaternary ammonium salt (cholestyramine powder) can reduce pancreatic Pancreatic fibrosis and pancreatic ductal malignant hyperplasia in cancerous mice;

Figure 8 shows that oral administration of polyaniline can reduce the activity of mTor pathway in pancreatic tissue, and inhibit the expression of Yap and Twist through autophagy. Figure 8 shows the results of Western blot analysis, including the expression of p21, Yap, P-Ttor, Twist, Cyclin D1, and actin in pancreatic tissue;

Figure 9 shows that oral polystyrylamine can reduce the mTor pathway in pancreatic tissue, resulting in the activation of autophagic flux. Histoimmunochemical staining experiments showed that LC3 accumulation and p62 accumulation in pancreatic tissue of pancreatic cancer mice increased autophagy-lysosome stress; on the contrary, cholestyramine can promote autophagy-lysosome stress in cancer tissue Flow, reduce LC3 and p62;

Figure 10 shows that oral administration of cholestyramine can reduce the expression of inflammatory factors in pancreatic tissue; RT-qPCR analysis of the expression of various inflammatory factors in mouse pancreatic cancer tissue; 6. The expression of TNF-alpha and Arg-1 suggests that oral administration of cationic polymers can reduce inflammation in cancer tissues by reducing bacterial toxins entering the blood, thereby inhibiting tumor growth and metastasis.

Detailed ways

The invention discloses the application of polycationic polymers in the preparation of medicines for treating or preventing tumors. Those skilled in the art can learn from the content of this article and appropriately improve the process parameters to realize it. In particular, it should be pointed out that all similar replacements and modifications are obvious to those skilled in the art, and they are all considered to be included in the present invention. The method and application of the present invention have been described through preferred embodiments, and the relevant personnel can obviously make changes or appropriate changes and combinations to the method and application described herein without departing from the content, spirit and scope of the present invention to realize and Apply the technology of the present invention.

In the present invention, the bacterial flagellin includes a variety of flagellin (flagellin) rich in acidic amino acids. Flagellin is a globular protein arranged in a hollow cylinder to form filaments in bacterial flagella . It has a mass of about 30,000 to 60,000 Daltons. Flagellin is the main component of bacterial flagella. After entering the blood, the immune system and inflammatory response of the body are activated by the pattern recognition receptor Toll-like receptor 5 (TLR5).

In the present invention, the CpG oligonucleotide (CpG oligodeoxynuleotide) is a short single-stranded DNA molecule, comprising a "cytosine triphosphate (cytosine)" and then a guanine triphosphate deoxynucleoside Acid (guanine). "P" refers to the phosphodiester chain between consecutive nucleotides, although some oligodeoxynucleotides have a modified phosphorothioate backbone. When these CpG motifs are unmethylated, they are pathogen-associated molecular patterns that, upon entering the bloodstream, stimulate the immune system by binding to the pattern recognition receptor Toll-like receptor 9 recognition (TLR9), leading to inflammation.

The research of the present invention shows that by taking orally administered large molecular weight and cationic polymers (large molecular weight and cationic polymers) that are not degraded or absorbed by the human body, by binding/neutralizing the endotoxin and other derivatives produced by intestinal microorganisms, It can eliminate intestinal bacterial toxins through stool, so as to reduce the acidic products of intestinal microorganisms into the blood. The invention can be applied to prevent, alleviate and improve tumors and cancers caused by intestinal bacterial toxins. The method can be used for preventing, mitigating, treating hyperbile acidemia and hyperbilirubinemia, and inhibiting tumors and their complications. The types of cancer mentioned here include but are not limited to: tumors and cancers of the digestive system, such as pancreatic cancer, liver cancer, rectal cancer, gastric cancer, and esophageal cancer. The application also includes the prevention, mitigation, and treatment of other tumors and cancers, such as bladder cancer, kidney cancer, lung cancer, breast cancer, head and neck cancer, skin cancer, and the like.

A large number of studies have shown that gut dysbiosis can promote the occurrence, development, and malignant metastasis of various tumors. The important consequence of intestinal flora disorder is the massive death of intestinal symbiotic bacteria; the large amount of endotoxin produced by them can enter the blood through the damaged intestine. In addition, gut microbes can also directly enter tumor tissue (intratumor microbes). Endotoxin entering the blood can cause systemic inflammation of the body, local tumor inflammation, tissue fibrosis, and tumor immune tolerance; this mechanism can damage and resist the immune monitoring system, resulting in the failure of tumor cells to be eliminated. Therefore, the invention can reduce intestinal toxins by taking cationic polymers that are not absorbed by the human body, such as Choestyramine, Colestipol, Colesevelam, etc. blood, thereby inhibiting systemic inflammation, alleviating the deterioration of various tumors, inhibiting tumor growth, and inhibiting tumor metastasis.

In the present invention, we provide a new application for the preparation of a drug for alleviating or treating pancreatic cancer. The application involves (1) alleviating the progression of pancreatic cancer and inhibiting the malignant transformation of pancreatic intraepithelial neoplasia (PanIN) to pancreatic ductal adenocarcinoma (PDAC); (2) inhibiting the local invasion and malignant systemic metastasis of pancreatic cancer; ( 3) Inhibit/relieve the complications of pancreatic cancer, such as jaundice; (4) Inhibit/relieve pancreatic fibrosis and liver fibrosis in cancer patients, and inhibit liver cirrhosis; (5) Reduce the content of endotoxin in the blood of cancer patients, reduce Toxemia, reduce elevated transaminases in the blood, improve liver function; (6) reduce various bile acid components in the blood of cancer patients, reduce bilirubin content in the blood, and relieve jaundice symptoms; (7) reduce cancer systemic inflammation storm, reduce systemic inflammation, reduce local inflammation in lesion tissue; (8) promote intestinal flora balance (eubiosis) in cancer patients. The above-mentioned treatment is to give patients a certain dose of high-molecular polycation compound in the form of oral administration, so as to improve any or all of the above-mentioned symptoms.

Cationic polymers, such as polystyrene quaternary ammonium salts or other cationic-rich organic polymers, which are not degraded by the intestinal tract, can effectively eliminate a variety of substances in the intestinal tract through their unique affinity including charge interaction. The endotoxin produced by negatively charged bacteria makes it excreted from the stool, thereby reducing the endotoxin entering the blood and reducing the endotoxin content in the blood. This application can relieve endotoxemia. A large number of studies have shown that a variety of endotoxins produced by intestinal microorganisms are an important source of inflammation in the body system. Chronic systemic inflammation may contribute to a variety of diseases, including diabetes, fatty liver, tumors, and cancer. Therefore, this application can alleviate the systemic inflammation caused by endotoxin and the local inflammation of the tumor environment, and improve the immune status of the tumor environment. One of the mechanisms applied by the invention is to reduce the endotoxin content in the blood, thereby reducing the AKT/mTOR pathway and inhibiting tumor growth. At the same time, the application of the invention can promote the increase of cell autophagy-lysosome flow, reduce the expression of oncogenes such as YAP, and reduce the state of cell stress. Taking this as the working mechanism and principle, the application of the invention can alleviate or inhibit the growth, transformation, deterioration and metastasis of tumor cells. In addition, the application of this right includes reducing endotoxin in the blood, thereby improving systemic inflammation, improving the pathological-physiological overall state of cancer patients, improving the mental state of patients, improving their quality of life, and possibly prolonging life. Because of its polymer properties and the property of not being degraded by the intestinal tract, it is not absorbed by the human body and can be excreted from the stool, with a high degree of safety and tolerance.

The application of the present invention also includes administering orally high-molecular cationic polymers to patients to remove other toxins produced by intestinal bacteria, such as CpG-DNA and bacterial flagellin. What they all have in common is anion-rich molecules. The endotoxin includes a variety of components, such as lipopolysaccharides (LPS) of bacterial cell walls, macromolecules composed of lipids and polysaccharides, including O antigens (O antigen), outer core and inner core. Endotoxins are mainly found in the outer membrane of Gram-negative bacteria. The term oligosaccharides refers to low molecular weight forms of bacterial lipopolysaccharides. Gram-negative bacteria in the gut come mainly from two main gates (phyla), Bacteroidetes and Proteobacteria. In addition, Enterobacter, Clostridium Botulinum, E.coli; Salmonella, Haemophilus Influenza, Vibrio, Klebsiella (Klebsiella) etc. are also an important source of intestinal toxins. In the intestinal environment, high-molecular cationic polymers can combine with endotoxins to make them excreted from the body.

The application of the present invention also includes giving patients oral gathering cationic resins to relieve jaundice in patients with pancreatic cancer and other complications by removing various bile acids (bile acids) and bilirubin (bilirubin) in the intestinal tract. disease. As an important complication in patients with pancreatic cancer, jaundice and bile acidemia can promote the growth of cancer cells. Oral administration of polystyrene quaternary ammonium salt (polyaniline, cholestyramine) or other pharmaceutical cationic resins with similar functions can reduce bile acids entering the blood and reduce the concentration of bile acids in tumor tissues. In this way, the AKT/mTOR signaling pathway in cancer cells can be inhibited to inhibit tumor growth. At the same time, this application can activate the autophagy-lysosome pathway, remove degenerated/damaged organelles in cells, decompose YAP, inhibit the growth of tumor cells, and inhibit the migration of tumor cells. In addition, through the application of this patent, the content of bile acids in the blood can be reduced, thereby improving systemic inflammation, improving the pathological-physiological overall state of cancer patients, improving the mental state of patients, improving their quality of life, and possibly prolonging their lives.

In the present invention, the medicine is a medicament prepared by adding an effective dose of polycationic resin as an active ingredient and adding pharmaceutically acceptable adjuvants or auxiliary ingredients. The preparation is an oral preparation.

The polycationic polymer is a polyamine polymer, and the polyamine polymer includes repeating units derived from the polymerization of amine monomers and crosslinking monomers; wherein, the preferred grade is the amine polymer It is an organic polymer containing tertiary amines and quaternary amines; the more preferred level is that the amine polymers are polystyrene quaternary ammonium salts, cholestyramine (cholestyramine, polyaniline, CAS: 11041-12-6) , Modified polyallylamine, copolyethyleneamine, polyallylamine, polylysine, etc. The most preferred level is that the said amine polymer is polystyrene quaternary ammonium salt, its chemical formula is C ₂₇ H ₄₇ N, which is a high molecular weight quaternary amine anion exchange polymer. The rights also include other polycationic polymers, such as Colestipol (Colestipol, CA: 37296-80-3), whose molecular formula is C ₈ H ₂₄ ClN ₅ , and Colesevelam, whose molecular formula is C ₃₁ H ₆₇ Cl ₃ N ₄ O, 2-(chloromethyl)oxirane; prop-2-en-1-amine; N-prop-2-enyldecan-1-amine; trimethyl-[6-(prop-2-enylamino )hexyl] azanium; chloride; hydrochloride. The original clinical indication application of these drugs is to clear intestinal bile acids, thereby lowering blood cholesterol levels. These macromolecular polymers are neither absorbed nor degraded in the intestinal tract, and their high safety performance is an important basis for the application of the invention.

The molecular weight of the high-molecular cationic polymer is greater than 4000 Da, preferably, the molecular weight of the high-molecular cationic resin is 1-10×10 ⁶ Da. The polymeric properties and the water insolubility of the polymer are structurally critical for this application, with safety and tolerance characteristics. The macromolecular polymer resin is not absorbed by the human body, and is excreted through the stool, taking away bacterial endotoxin, bile acid and other components at the same time.

Described macromolecule cationic polymer has:

(1) With multiple positive charges;

(2) The skeleton is an organic covalent bond polymer polymer;

(3) The cationic polymer is not decomposed by the enzymes of the digestive tract, can be excreted through the digestive tract, and will not be decomposed and absorbed by the body.

The high-molecular cationic polymer provided by the present invention removes or neutralizes the application of pathogenic factors produced by intestinal microorganisms, and the intestinal pathogenic factors include but not limited to: endotoxin (lipopolysaccharide), negatively charged long Chain/short chain fatty acids (including acetic acid, propionic acid, butyric acid), hydrogen sulfide, negatively charged microbial derivatives and decomposition products, bile acids, and bacterial DNA fragments (CpG DNA) and nucleotides ,etc.

Preferably, the positively charged high molecular weight polymer is administered orally to neutralize intestinal microbial endotoxins. On the contrary, our animal experiments showed that the uncharged polymer hollow resin has no therapeutic effect. Therefore, we prove one of the structural importance of the positive charge in its structure; at the same time, the hydrophobic structure of its polymer is also the key point of the patented invention. The high molecular weight amine polymer is polystyrene quaternary ammonium salt, cholestyramine (cholestyramine, polyaniline), colestipol (Colestipol, Colestipol), colesevelam (Colesevelam) or similar cation-rich High molecular polymer, used to alleviate, prevent and treat a variety of diseases caused by endotoxin entering the blood, including metabolic fatty liver disease, alcoholic fatty liver disease (alcoholic fatty liver diseases), non-alcoholic fatty liver disease (non-alcoholic fatty liver disease) liver diseases, NAFLD), type 2 diabetes.

In the application of the high-molecular cationic polymer provided by the present invention in the preparation of drugs for treating or preventing tumors, the raw materials and reagents used can be purchased from the market.

Below in conjunction with embodiment, further set forth the present invention:

Example 1

The preparation and clinical application of the high-molecular cationic polymer of the present invention include treating, alleviating and preventing pancreatic cancer and other cancers, including their various complications. As an example of implementation, it includes amine polymer; its preferred practical application is polystyrene quaternary ammonium salt, cholestyramine (cholestyramine, referred to as "polyaniline"). Repeating units derived from the polymerization of monomers, such as polystyrene quaternary ammonium salt ([4-[3-(4-ethylphenyl)butyl]phenyl]-trimethylazanium), its molecular formula is C ₂₁ H ₃₀ N+, including its similar Various derivative structures, such as modified polyallylamine, copolymer of diethylenetriamine, (C ₄ H ₁₀ N ₃ ) _m (C ₃ H ₆ O) _n ). Its molecular mass (molecular mass) exceeds 1x10 ⁶ g/mol. The general formula of its molecular structure is as follows:

The cationic resin can be taken orally alone, or added to food, and taken in the form of meals. Dosage, for adults, can be 1-8 grams/time, 1-3 times a day, the maximum daily dose is 30 grams. The specific dosage and duration of treatment should be determined according to the Pharmacopoeia and the prescriptions of medical personnel, within a reasonable range, according to the severity of the disease, and the guidelines of drug regulatory agencies.

Example 2

The preparation and clinical application of the high-molecular cationic polymer of the present invention include treating, alleviating and preventing pancreatic cancer and other cancers, including the complications of these cancers. As an example of implementation, amine polymers are included. As the preferred application, Colestipol (Colestipol, hydrochloride, Epichlorhydrin-tetraethylenepentamine polymer, CAS No.37296-80-3; SDS CAS No:37296-80-3; Molecular Weight: (225.765) n; Molecular Formula: C ₈ H ₂₄ CLN5), or similar cation-rich polymers.

Its molecular mass (molecular mass) exceeds 1x10 ⁶ g/mol. The general formula of its molecular structure is as follows:

Colestipol molecular formula: (C ₄ H ₁₀ N ₃ ) _m (C ₃ H ₆ O) _n .

Its molecular structure general formula is as follows:

Therefore, the high-molecular-weight cationic polymer can be used for the treatment, prevention of said diseases, and the alleviation of the related symptoms produced by these diseases. For example, polymer cationic resin can be used for the prevention and treatment of liver fibrosis caused by non-alcoholic steatohepatitis, liver cirrhosis.

The high-molecular cationic polymer can be taken orally alone, or added to food, and taken in the form of meals. Dosage, for adults, can be 1-8 grams/time, 1-3 times a day, the maximum daily dose is 30 grams. The specific dosage and duration of treatment should be determined according to the Pharmacopoeia and the prescriptions of medical personnel, within a reasonable range, according to the severity of the disease, and the guidelines of drug regulatory agencies.

Example 3

Colesevelam is another high-molecular cationic polymer, and its bile acid binding capacity is 7 times that of cholestyramine. Based on the similarity of its mechanism, we speculate that oral administration of colesevelam can also reduce the endotoxin produced by intestinal cells and the endotoxin in the blood, thereby reducing the systemic inflammation of the body, restoring insulin sensitivity, reducing fatty liver, and promoting Self-ablation of NASH and cirrhosis. The daily dosage can be viewed as 3.75g, which can be administered in powder or tablet form. The molecular formula of colesevelam is C ₃₁ H ₆₇ Cl ₃ N ₄ O ; the International Union of Pure and Applied Chemistry (IUPAC) named it: 2-(chloromethyl)oxirane; prop-2-en-1-amine; N-prop -2-enyldecan-1-amine; trimethyl-[6-(prop-2-enylamino)hexyl]azanium; chloride; hydrochloride.

Example 4

The application core of the invention is to prevent the deterioration of tumors, relieve the growth of tumors and reduce the migration of tumors by taking cationic polymers. Taking cationic polymers, such as polystyrene quaternary ammonium salt (cholestyramine, Cholestyramine), can reduce endotoxin/bile acid entering the blood and promote tumor growth by clearing out endotoxin produced by intestinal microorganisms and intestinal bile acid. Autophagy of cells, relief of lysosomal stress, removal of damaged/degenerative organelles in cells, etc., to inhibit the growth and metastasis of tumor cells. At the same time, taking cationic polymers can also reduce the overall systemic inflammation of the body, alleviate the degree of inflammation in the tumor environment, improve the tumor immune status, and inhibit tumor metastasis. As an example of a specific application, the amount of polystyrene quaternary ammonium salt (cholestyramine powder) can be used in a certain range, such as 1-8 grams per time, 1-3 times a day, and the maximum daily dose is below 30 grams. As an application example, it also includes taking colesevelam at a dose of 1-8 grams per day; or colestipol at a dose of 1-15 grams per day. The specific dosage and duration of treatment should be determined according to the Pharmacopoeia and the prescriptions of medical personnel, within a reasonable range, according to the severity of the disease, and the guidelines of drug regulatory agencies.

Example 5

The application of the invention also includes the combined application of taking cationic polymers and other treatment methods, including combined application with tumor removal surgery, anti-tumor chemotherapy and anti-tumor immunotherapy. Its specific scope of use covers the reasonable range of application of the original therapy.

Example 6 Administering high-molecular cationic polymers can inhibit the growth of pancreatic cancer.

6-week-old WT (C57BL/6J) mice and 6-week-old pancreatic cancer KC (PDX1-KrasG12D) mice were divided into three groups: (1) WT mice were given maintenance diet (blank control group n=10); (2) ) pancreatic cancer KC mice were given maintenance feed (control group n=6); (3) pancreatic cancer KC mice were given maintenance feed containing 3% cholestene (experimental group n=6); continuous feeding for 10 weeks. As shown in Figure 1, the pancreas lesions and pathological changes of mice in each group were counted at the end of the experiment. As shown in Figure 1A, taking a certain amount of polystyrene quaternary ammonium salt can effectively inhibit the growth of pancreatic cancer in the three time periods of "early-middle-late". Figure 1B shows the histology of pancreatic cancer. Under the action of PDX-Kras, pancreatic cancer and pancreatic ductal carcinoma were born with a large number of hyperplasia, and adding a certain dose of polystyrene quaternary ammonium salt to the feed can effectively inhibit the proliferation of tumors in the pancreas .

Example 7 Administering high-molecular cationic polymers can inhibit the progression of pancreatic cancer metastasis.

6-week-old WT (C57BL/6J) mice and 6-week-old pancreatic cancer KC (PDX1-KrasG12D) mice were divided into three groups: (1) WT mice maintained diet (blank control group n=10); (2) Pancreatic cancer KC mice were given maintenance diet (control group n=6); (3) pancreatic cancer KC mice were given maintenance diet containing 3% cholestyramine (experimental group n=6); feeding continued for 10 weeks. Figure 2 shows the metastases of pancreatic tumors. In the PDX-Kras genetic background, tumor metastasis is mainly manifested in tumor rectal metastasis; the pancreatic cancer metastasis in the illustration also includes intraocular and other visceral metastasis. High-molecular cationic polymer (cholestyramine) can reduce the rate of cancer metastasis from 63% of the control to 36%. As shown in the figure, the state of biliary congestion was also significantly improved.

Example 8 Biliary obstruction and jaundice promote the development of pancreatic cancer

Patients with pancreatic cancer are often accompanied by complications such as biliary obstruction and jaundice. To verify its causality, we used bile duct ligation. Nine-week-old pancreatic cancer KC (PDX1-KrasG12D) mice were divided into two groups: (1) mice were given sham operation of common bile duct ligation (control group n=12); (2) mice were given common bile duct ligation, mice Common bile duct obstruction was formed, and they were sacrificed after two weeks of continuous feeding (n=15). Figure 3A is a schematic diagram of the experimental design. Figure 3B shows the histobiological status of mouse pancreatic lesions. As shown, bile duct ligation promotes pancreatic cancer growth and metastasis.

Example 9 Bacterial endotoxin (LPS) can promote the development of pancreatic cancer

Endotoxins produced by gut bacteria are an important contributor to systemic inflammation as well as local inflammation in the tumor environment. Here, we determined whether endotoxin could aggravate pancreatic cancer in mice with a PDX-Kras genetic background. Here, 9-week-old pancreatic cancer KC (PDX1-KrasG12D) mice were divided into two groups: (1) mice were given intraperitoneal injection of 100 microliters of normal saline (control group n=12); (2) mice were given 4 mg/ kg Escherichia coli LPS was injected intraperitoneally once a week for four weeks and the mice were harvested. Figure 4A is a schematic diagram of the experimental design. Figure 4B is staining of pancreatic tissue. HE staining showed that administration of LPS could aggravate the growth of pancreatic cancer, while Masson Trichrome staining showed that LPS could aggravate the fibrosis of pancreatic cancer tissue. Figure 4C is the immunochemical staining of pancreatic tissue; Ck19 indicates the proliferation of ductal carcinoma, K67 indicates the proliferation of tumor cells, and Collagen type-1 indicates tissue fibrosis.

Example 9 The increase of blood bile acid in mice with pancreatic cancer can be alleviated by taking high-molecular-weight cationic polymers

Due to the growth of pancreatic tumors, bile duct compression and cholestasis are often caused, resulting in hyperbiliary acidemia and jaundice, which can promote tumor transformation and malignancy. To confirm this speculation, we used 6-week-old C57BL6J mice and pancreatic cancer mouse KC (PDX1-KrasG12D) mice of the same age, and divided them into the following three groups: (1) WT (C57BL/6J) mice were given to maintain feed (blank control group n=6) feeding; (2) giving pancreatic cancer KC (PDX1-KrasG12D) mice maintenance feed feeding (control group n=13); (3) giving pancreatic cancer KC (PDX1-KrasG12D) mice The maintenance diet containing 3% cholestene was fed (experimental group n=13) for 10 weeks. As shown in Figure 5A, the content of bile acid in the serum of mice with pancreatic cancer was significantly higher than that of the control WT; at the same time, as shown in Figure 5B, administration of polystyrene quaternary ammonium salt could significantly improve the level of serum total bile acid.

Example 10 Taking high-molecular cationic polymers can effectively reduce the endotoxin content in the blood of mice

The key point of the invention is to alleviate, prevent and reduce pancreatic cancer by using cationic aggregate polymer. Second, we speculate that administration of cationic resins can effectively remove endotoxins in the gut, which is an important cause of many chronic diseases, including cancer. 6-week-old WT (C57BL/6J) mice and 6-week-old pancreatic cancer KC (PDX1-KrasG12D) mice were divided into three groups: (1) WT mice were given maintenance diet (blank control group n=10); (2 ) KC mice with pancreatic cancer were given maintenance feed (control group n=6); (3) KC mice with pancreatic cancer were given maintenance feed containing 3% cholestyramine (experimental group n=6); feeding continued for 10 weeks. As shown in Figure 6, at the end point of the experiment, the level of LPS in the serum, administration of cholestene quaternary ammonium salt can effectively reduce the level of endotoxin in the serum of mice with pancreatic cancer, suggesting that endotoxin can be reduced by excreting intestinal endotoxin. .

Example 11 Administering high-molecular cationic polymers can reduce pancreatic fibrosis and malignant hyperplasia of pancreatic ducts in mice with pancreatic cancer

Tissue fibrosis is an important cause of pancreatic cancer progression and an important basis for the tumor microenvironment. Sustained tissue damage and chronic inflammation are important causes of tissue fibrosis. Therefore, eliminating fibrosis and clearing the tumor environment is a strategy for tumor therapy. We speculate that clearing intestinal endotoxins may reduce persistent inflammation in cancer tissues, thereby eliminating fibrosis of tumor tissues and inhibiting tumor proliferation. For this reason, 6-week-old WT (C57BL/6J) mice and 6-week-old KPC (PDX1-KrasG12D) mice were divided into three groups: (1) WT mice were given maintenance feed (blank control group n=10); ( 2) Pancreatic cancer KC mice were given maintenance diet (control group n=30); (3) Pancreatic cancer KC mice were given maintenance diet containing 3% cholestyramine (experimental group n=28) for 10 weeks. Figure 7A shows fibrosis (Masson staining) and ductal carcinoma growth status (CK-19) in pancreatic tissue. Figure 7B shows semi-quantitative scanning data, showing that taking polystyrene quaternary ammonium salt (cholestyramine powder) can reduce pancreatic fibrosis and pancreatic ductal malignant hyperplasia in mice with pancreatic cancer

Example 12 Taking high-molecular cationic polymers can reduce the mTor pathway in pancreatic tissue, promote cell autophagy to inhibit the expression of Yap and Twist

A large number of studies have shown that growth factors can promote cell growth metabolism and multiple anabolism by activating the mTOR pathway of cells. Similarly, activation of the mTOR pathway can inhibit cell autophagy (autophagy), thereby reducing the cell's self-clearance ability, resulting in the accumulation of damaged organelles, lysosome stress, so that YAP cannot be effectively cleared, and YAP is a important tumor factor. Figure 8 shows the results of Western Blot analysis of the pancreatic tissues of each group of mice at the end of the experiment. As shown in the figure, administration of polystyrene quaternary ammonium salt (cholestyramine powder) can increase the expression of p21 in pancreatic cancer tissue, suggesting that its mechanism of tumor suppression is activated; at the same time, administration of polystyrene quaternary ammonium salt can Inhibition of mTor, promotion of autophagy (Atg7 and reduction of p62), resulting in a decrease in Yap. As shown in Figure 9, at the level of pancreatic tissue, the administration of polystyrene quaternary ammonium salt can reduce the expression of p62 in tumor tissue, suggesting that its autophagy is accelerated, and Yap is degraded through lysosome.

Example 13 Oral administration of high-molecular cationic polymers can reduce the expression of inflammatory factors in pancreatic cancer tissue

The disturbance of intestinal flora, the production of endotoxin and the damage of intestinal tight junction can lead to the entry of endotoxin into the blood, thereby generating systemic inflammation and local inflammation in tumor tissue. Persistent inflammation can promote cellular oxidative stress (ROS, reactive oxidative species), which can produce cell damage, gene mutation, cell transdifferentiation, cell aging, and promote cancer at the genetic and epigenetic levels. At the same time, inflammatory factors can also promote tissue damage, trigger tissue proliferation, and fibrosis. Here, we determined the expression status of pancreatic inflammatory factors in pancreatic cancer KC (PDX1-KrasG12D) mice given cholestyramine treatment and control mice. As shown in FIG. 10 , administering polystyrene quaternary ammonium salt (cholestyramine) to mice with pancreatic cancer can significantly inhibit the expression of multiple inflammatory factors in pancreatic cancer tissue. For example, taking polystyrene quaternary ammonium salt (cholestyramine powder) can inhibit the expression of interleukin-6 (Interleukin-6) and tumor necrosis factor-alpha (TNF-alpha) in pancreatic cancer tissue. At the same time, the expression of Arginase-2 is a sign of deep transformation of inflammation, indicating the transformation of Th2 and M2, and is an inflammatory indicator of long-term tumor development. Here, we found that the administration of polystyrene quaternary ammonium salt (cholestyramine powder) can inhibit the expression of Arginase-2 in pancreatic cancer tissues. Therefore, the inhibition of pancreatic cancer by taking polystyrene quaternary ammonium salt (cholestyramine powder) is accomplished to some extent by improving the local inflammatory state of the tumor.

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

The application of the cationic polymer in the preparation of preparations or medicines for eliminating pathogenic factors produced by intestinal microorganisms.
The application according to claim 1, wherein the pathogenic factors include but are not limited to: intestinal endotoxin (endotoxin, lipopolysaccharide), short-chain fatty acids (including acetic acid, propionic acid, butyric acid), hydrogen sulfide, Negatively charged microbial derivatives and decomposition products, bile acids, intestinal microbial DNA fragments (CpG-DNA), enterovirus DNA and RNA, various nucleotides (nucleotides), bacterial flagellin ( flagellin).
Application of the oral cationic polymer in the preparation of preparations or medicines for removing various acidic derivatives produced by intestinal microorganisms.
Application of the oral cationic polymer in the preparation of preparations or medicines for preventing, alleviating, improving and/or treating diseases caused by derivatives produced by intestinal microorganisms and various bacterial toxins.
The application according to claim 4, wherein the diseases include tumors, systemic inflammation of the body, local inflammation of tumors, tissue fibrosis, and tumor immune tolerance;

Diseases caused by systemic inflammation also include diabetes and fatty liver.

Oral administration of the preparation or drug can reduce the endotoxin content in the blood, thereby reducing the cellular AKT/mTOR pathway, activating the cellular autophagy-lysosome pathway, to remove degenerated/damaged organelles in cells, decomposing YAP, and inhibiting tumor cells Growth of tumor cells, inhibition of tumor cell migration, promotion of autophagy-lysosomal flux, reduction of oncogene expression and/or reduction of cell stress state.
The application according to any one of claims 1 to 5, the cationic polymer comprises the following features:

(1) Multiple positive charges; and

(2) The skeleton is an organic covalent bond polymer polymer;

Preferably, the high-molecular cationic polymer also includes a hydrophobic structure.
The application according to claim 6, characterized in that, the high-molecular cationic polymer comprises a polyamine polymer; the polyamine polymer comprises repeating units derived from the polymerization of amine monomers and crosslinking monomers ;

The polyamine polymer is an organic polymer containing tertiary amine or quaternary amine.
The application according to claim 7, wherein the cationic polymer comprises polystyrene quaternary ammonium salt, cholestyramine (cholestyramine, polyaniline, CAS: 11041-12-6), modified polypropylene Polyvinylamine, polyvinylamine polyallylamine, DEAE-cellulose, polymyxin B-crosslinked polystyrene or cellulose or dextran resin (polymyxin B-conjugated polystyrene/fibers/agarose), polyethylene Amines, cationic starch or cellulose, chitosan and its derivatives, etc., polypeptides or proteins rich in lysine/arginine; most preferably, the amine polymer includes Choestyramine , Colestipol (Colestipol), Colesevelam (Colesevelam); the molecular weight of the high-molecular cationic polymer is greater than 6000Da to avoid entering the body, can be completely discharged from the stool, and bring out a variety of toxins; the The high molecular weight cationic polymer has a molecular weight of 1×10 6 to 10×10 6 Da.
The application according to claim 7, characterized in that, by oral administration of cholestyramine or colestipol or colesevelam, to remove endotoxins produced by intestinal bacteria; endotoxins include lipopolysaccharides (lipopolysaccharides) of bacterial cell walls , LPS), or other toxins including bacterial CpG-DNA, bacterial flagella. To prevent, alleviate and treat various cancers, including one or more of pancreatic cancer, liver cancer, rectal cancer, gastric cancer, esophageal cancer, bladder cancer, kidney cancer, lung cancer, breast cancer, head and neck cancer, and skin cancer;
The application according to claim 9, wherein the prevention, alleviation, improvement and/or treatment of pancreatic cancer is to alleviate the following symptoms:

(1) Relieve the progression of pancreatic cancer and inhibit the malignant transformation of pancreatic intraepithelial neoplasia (PanIN) to pancreatic ductal adenocarcinoma (PDAC); and/or

(2) Inhibit local invasion and malignant systemic metastasis of pancreatic cancer; and/or

(3) Inhibition/remission of complications of pancreatic cancer, such as jaundice; and/or

(4) Inhibit/relieve pancreatic fibrosis and liver fibrosis, inhibit liver cirrhosis in cancer patients; and/or

(5) Reduce the endotoxin content in the blood of cancer patients, reduce endotoxemia, reduce the increase of transaminase in the blood, and improve liver function; and/or

(6) Reduce the various bile acid components in the blood of cancer patients, reduce the bilirubin content in the blood, and relieve the symptoms of jaundice; and/or

(7) reduce cancer systemic inflammatory storm, reduce systemic inflammation, reduce local inflammation in lesion tissue; and/or

(8) Promote the intestinal flora balance (eubiosis) of cancer patients.