Use of a lariat peptide for the preparation of a medicament for alleviating doxorubicin cardiotoxicity

Abstract

The application discloses a use of a larazotide polypeptide in preparation of a drug for relieving adriamycin cardiotoxicity. In-vivo experimental results show that the larazotide can effectively relieve the toxic effect of a chemotherapeutic drug adriamycin on the heart, relieve myocardial enzyme spectrum increase, enhance left ventricular diastolic capacity and cardiac ejection fraction by improving body and heart immune cell homeostasis and reducing myocardial tissue cell apoptosis, thereby reducing the toxic side effect of the chemotherapeutic drug adriamycin, and improving intestinal barrier function damage caused by adriamycin chemotherapy. The application overcomes the defects of current clinical drug limitations of Dox and existing chemotherapeutic heart protection drugs, and solves the prevention and treatment problems of heart toxicity and intestinal side effects induced in Dox chemotherapy.

Description

Technical Field

[0001] This invention relates to a novel use of a larizole polypeptide, specifically to the use of a larizole polypeptide in a drug for alleviating doxorubicin cardiotoxicity. Background Technology

[0002] Doxorubicin (Dox) is a broad-spectrum, highly effective anthracycline anticancer drug commonly used in clinical practice, widely applied in the treatment of leukemia, malignant lymphoma, breast cancer, ovarian cancer, and osteosarcoma. Although anthracyclines, represented by Dox, have increased the long-term survival rate of cancer patients, their dose-dependent cardiotoxicity and the potential for irreversible myocardial damage and congestive heart failure significantly limit their clinical application.

[0003] Currently, cardioprotective strategies for preventing anthracycline toxicity mainly include using cardioprotective agents or reducing the cumulative dose of chemotherapy drugs to mitigate cardiotoxic effects. Dextromethorphan is currently the only approved cardioprotective agent against anthracycline toxicity. Although dextromethorphan, when used in combination with anthracycline chemotherapy drugs, can prevent a decrease in resting left ventricular ejection fraction and reduce the incidence of heart failure, it can exacerbate chemotherapy-induced myelosuppression, affecting antitumor efficacy. Furthermore, clinical studies have shown that using the FAC regimen (fluorouracil, doxorubicin, cyclophosphamide) plus dextromethorphan may result in patients receiving a higher cumulative dose of doxorubicin (compared to the group without dextromethorphan), but it does not eliminate anthracycline-induced cardiotoxicity. In addition, treatment with large doses of reactive oxygen species (ROS) scavengers and iron chelators is not effective in preventing Dox-induced cardiomyopathy in cancer patients or has significant clinical application limitations. Due to these problems in current clinical drug use, the development of novel drugs and interventions to treat anthracycline chemotherapy cardiotoxicity is urgently needed. In clinical practice, Dox chemotherapy can irritate the gastric mucosa, causing symptoms such as nausea, vomiting, abdominal distension, abdominal pain, or constipation. While probiotic supplementation can alleviate these symptoms and aid digestion, it does not alleviate Dox-induced cardiotoxicity. Dox treatment can also damage intestinal epithelial function, Paneth cells, and mucus, leading to intestinal barrier mucosal damage. The intestinal barrier maintains intestinal homeostasis and prevents the translocation and release of pathogens and toxins into extraluminal tissues and organs. Damage to the intestinal barrier allows for the absorption and migration of large amounts of bacteria and endotoxins into the bloodstream and lymphatic system, potentially causing systemic multi-organ dysfunction, including sepsis, vascular and cardiac remodeling. Studies have found that activated intestinal immune cells during intestinal barrier damage migrate and cause inflammation in distant organs. We hypothesize that in addition to the direct effects of Dox on the heart via circulation, the resulting intestinal barrier damage and subsequent imbalance of immune cell homeostasis in the body and heart may be a significant factor exacerbating Dox-induced cardiotoxicity.

[0004] Based on the above viewpoints, this invention uses the biomolecule Larazotide, which has intestinal barrier protection function, to intervene in advance in the early stage of Dox treatment, thereby maintaining the integrity of the intestinal barrier during Dox chemotherapy, and further improving the imbalance of the body's and heart's immune cell homeostasis, reducing the cardiotoxicity risk of Dox chemotherapy.

[0005] Larazotide is a biomacromolecule with potential intestinal barrier protection.

[0006] Larazotide is an octameric peptide and its acetate derivative, larazotide acetate, that can inhibit early mucosal events leading to barrier dysfunction and immune activation, and has therapeutic value for celiac diseases. Larazotide inhibits gliadin-induced macrophage accumulation in the intestine and maintains the normal structure of tight junction proteins (TJs), promoting the recovery of intestinal barrier damage.

[0007] Although larazotide has shown potential biological functions in improving gastrointestinal diseases, its role and protective mechanism in alleviating cardiotoxicity caused by Dox chemotherapy for tumors have not been reported.

[0008] This invention provides a potential drug and intervention method to reduce the cardiotoxicity of doxorubicin chemotherapy, while also improving the intestinal function damage caused by doxorubicin. Furthermore, due to the convenience of the above-mentioned intervention method, this invention can effectively reduce the pain, psychological and economic burden of chemotherapy for cancer patients. Summary of the Invention

[0009] The purpose of this invention is to overcome the shortcomings of existing drugs and technologies and provide a use of larazotide in alleviating doxorubicin-induced cardiotoxicity. This invention is the first to propose the prevention and treatment of doxorubicin (Dox) chemotherapy-induced cardiotoxicity through oral administration of larazotide, overcoming current restrictions on the clinical use of Dox and the drawbacks of existing chemotherapy-protective drugs, while simultaneously solving the problem of preventing and treating Dox-induced cardiotoxicity.

[0010] The objective of this invention is achieved through the following technical solution:

[0011] This invention relates to the use of a larizoletine polypeptide or a derivative thereof in the preparation of a drug to alleviate the cardiotoxicity of doxorubicin chemotherapy.

[0012] The larazotide polypeptide (CAS: 258818-34-7) has the molecular formula C 32 H 55 N9O 10The amino acid sequence is: glycylglycyl-L-valine-L-leucyl-L-valine-L-glutamine-L-prolylglycine.

[0013] As one embodiment of the present invention, the larizolide polypeptide derivative includes larizolide polypeptide acid salt derivative.

[0014] As one embodiment of the present invention, the larizolide peptide is used as follows: once a day starting one week before Dox treatment, and once every other day after Dox treatment.

[0015] As one embodiment of the present invention, the dosage of larazotide for mice is as follows: prepare a stock solution of larazotide with a concentration of 1 mg / mL, and administer 0.2 mg / day per mouse by gavage.

[0016] As one embodiment of the present invention, the dosage of larizole polypeptide for adults is: 0.0385g to 0.154g / day of oral administration per 70kg body weight.

[0017] Compared with the prior art, the present invention has the following beneficial effects:

[0018] 1. This invention, by administering larazotide orally via gavage one week prior to chemotherapy, can effectively prevent and treat cardiotoxicity caused by doxorubicin (Dox) chemotherapy, including significantly improving left ventricular systolic and diastolic function, increasing ejection fraction, reducing circulating myocardial enzyme levels, and lowering cardiac fibrosis and apoptosis levels; it also has the effect of protecting against chemotherapy-induced intestinal barrier damage.

[0019] 2. Larazotide is a phase III clinical trial drug, mainly indicated for intestinal diseases such as enteritis and celiac disease. From a clinical application perspective, it is safer than other drugs for preventing and treating Dox chemotherapy cardiotoxicity.

[0020] 3. The mechanism by which Larazotide alleviates Dox chemotherapy-induced cardiotoxicity is as follows:

[0021] 3.1 Declaration can regulate the structure and expression level of tight junction protein (TJ) in the intestinal epithelial barrier, and promote the barrier recovery of the intestinal barrier under acute and chronic Dox injury.

[0022] 3.2 Larazotide restores the integrity of the intestinal barrier and can effectively reduce endotoxemia or bacterial translocation caused by intestinal barrier damage. It is important for protecting against the entry of Dox-induced endotoxins into the circulation and their impact on cardiac function.

[0023] 3.3 This invention found that in Dox mice treated with Larazotide, there were significant differences in the Th1 and Th2 responses. IFN-γ (Th1) levels decreased significantly, while IL-4 (Th2) levels remained unchanged. Flow cytometry analysis showed a significant decrease in the proportion of cardiac T cells. Larazotide alleviates Dox cardiotoxicity by improving intestinal barrier function, reducing the accumulation of circulating immune cells, maintaining immune cell homeostasis in the body and heart, and mitigating the excessive response of T cells to myocardial tissue, thereby inhibiting cardiomyocyte apoptosis. Attached Figure Description

[0024] Other features, objects, and advantages of the present invention will become more apparent from the following detailed description of non-limiting embodiments with reference to the accompanying drawings:

[0025] Figure 1 The results of left ventricular echocardiography in mice after Larazotide intervention;

[0026] Figure 2 Serum myocardial enzyme levels in mice after Larazotide intervention;

[0027] Figure 3 Pathological tissue sections were analyzed from mouse hearts (Masson and TUNEL) and intestines (H&E);

[0028] Figure 4 The data represent circulating IFN-γ and IL-4 levels and the composition of immune cells in the heart; where A represents plasma IFN-γ and IL-4 levels, and B represents the analysis of cardiac immune cell composition. Detailed Implementation

[0029] The present invention will be described in detail below with reference to embodiments. These embodiments will help those skilled in the art to further understand the present invention, but do not limit the invention in any way. It should be noted that those skilled in the art can make several adjustments and improvements without departing from the concept of the present invention. These all fall within the protection scope of the present invention.

[0030] Example 1

[0031] 1. Laboratory animals and grouping

[0032] SPF-grade male C57BL / 6 mice, aged 6-8 weeks and weighing approximately 18-20g, were purchased from Beijing Spaford Laboratory Animal Co., Ltd. and housed at the Experimental Animal Center of Zhengzhou University.

[0033] The experimental groups included: oral PBS group (PBS group), oral PBS + Dox injection group (Dox group), oral larazotide group (LA group), and oral larazotide + Dox injection group (LA+Dox group).

[0034] 2. Handling methods

[0035] PBS group and Dox group: The experiment was conducted one week before the first Dox injection. Both groups were given 200 μl of PBS by gavage daily. After the first Dox injection, the gavage was continued every other day until the end of the experiment.

[0036] The LA group and the LA+Dox group: The experiment was conducted one week before the first Dox injection, with Larazotide 200μl administered by gavage daily. After the first Dox injection, the gavage was continued every other day until the end of the experiment.

[0037] After three Dox injections are completed, observe for 3-4 weeks before proceeding with subsequent experimental procedures.

[0038] 3. Modeling methods

[0039] Dox-induced chronic cardiotoxicity mouse model: Mice were injected intravenously with Dox at a final concentration of 15 mg / kg in three separate doses, 5 mg / kg each time, with an interval of 7 days. After establishing the model, the mice's food intake, mental state, and activity response were closely observed. The mice were weighed the day after model establishment.

[0040] 4. Small animal ultrasound

[0041] Left ventricular cardiac function changes in mice were monitored at regular intervals before Dox injection and every week after injection. When significant cardiac function changes were observed, the mice were euthanized and intestinal, blood, and heart samples were collected.

[0042] 5. Pathological examination of intestinal and cardiac tissues

[0043] A 2cm segment of the distal ileum was harvested, rapidly rinsed with cold saline, and the intestinal contents were removed. The segment was then fixed in a tissue fixative for 24-48 hours. After H&E staining, the morphological changes of the intestinal tissue were observed under a light microscope, and a pathological score was performed.

[0044] Mouse heart tissue was collected, and after removing blood from inside and outside the heart cavity, it was fixed in tissue fixative for 24-48 hours. H&E, Masson, and TUNEL staining were used to analyze changes in heart tissue structure, the degree of myocardial fibrosis, and the level of cardiomyocyte apoptosis.

[0045] 6. Monitoring of serum myocardial enzyme profile and intestinal barrier integrity

[0046] The expression levels of myocardial enzymes (CKMB, cTnT, NTpro-BNP) in each group of mice were analyzed using ELISA to confirm the differences in cardiac function among the groups. The morphology of intestinal tissue in each group of mice was analyzed to assess the improvement of the intestinal barrier.

[0047] 7. Results

[0048] 7.1 Growth status:

[0049] PBS group: Mice had normal food intake and defecation, good activity levels, increased body weight during the experiment, and no deaths occurred;

[0050] Dox group: Within 3 days after the first two Dox injections, mice showed varying degrees of decreased activity, slow movement, curled-up body, dull and bristly fur, and significant weight loss. After 3 days, their weight gradually recovered and their activity level slowly recovered. However, after the third and fourth Dox injections, their activity level decreased significantly, and their weight remained or decreased.

[0051] LA group: Rats had normal food intake and defecation, good activity levels, increased body weight during the experiment, and no deaths occurred;

[0052] LA+Dox group: Mice experienced a slight decrease in body weight within 3 days after Dox injection, followed by recovery and maintenance of body weight; activity level did not decrease significantly.

[0053] 7.2 Improvement in cardiac function ( Figure 1 , 2 ):

[0054] PBS group: The left ventricular function of mice was normal during the experimental stage, myocardial enzyme levels were normal, myocardial tissue showed no obvious fibrosis, and myocardial cells showed no obvious apoptotic changes.

[0055] Dox group: After mice were injected with three doses of Dox, with a cumulative dose of 15mg, left ventricular function began to decline, serum myocardial enzyme expression increased significantly, myocardial fibrosis worsened, and myocardial cell apoptosis was obvious.

[0056] LA group: The left ventricular function of mice was normal during the experimental stage, myocardial enzyme levels were normal, myocardial tissue showed no obvious fibrosis, and myocardial cells showed no obvious apoptotic changes.

[0057] LA+Dox group: After mice were injected with three doses of Dox, with a cumulative dose of 15mg, there was no significant decrease in left ventricular function, the expression of serum myocardial enzymes was significantly lower than that in the Dox group, there was no obvious myocardial fibrosis, and myocardial cell apoptosis was not significant.

[0058] Figure 4 Composed of circulating IFN-γ and IL-4 levels and immune cells in the heart; Figure 4It was found that there were significant differences between the Th1 and Th2 responses in mice after Larazotide intervention. The Th1 response decreased significantly and returned to normal levels, while the Th2 response remained unchanged. Simultaneously, the composition of immune cells, especially T cells, in the cardiac tissue was significantly restored compared to the Dox group, indicating that Larazotide can reduce Dox-induced T cell damage to the myocardium, thereby alleviating the cardiotoxic effects of Dox.

[0059] 7.3 Improvement of the intestinal barrier ( Figure 3 ):

[0060] PBS group: The model endpoint showed that the mouse intestinal tissue morphology was normal, the ileum inflammatory cell infiltration was not obvious, the villi were normally stretched, and the levels of LPS and Zonulin were normal.

[0061] Dox group: Model endpoint, mouse intestinal tissue was more brittle, inflammatory cell infiltration in the ileum was obvious, villi were broken, LPS and Zonulin levels were significantly increased, and the expression of intestinal epithelial tight junction proteins ZO-1, Occludin, and Cludin-1 was significantly decreased.

[0062] LA group: Model endpoint, normal morphology of mouse intestinal tissue, no obvious inflammatory cell infiltration in the ileum, normal villus spreading, and normal levels of LPS and Zonulin;

[0063] LA+Dox group: The model endpoint showed that the intestinal tissue morphology of mice was normal, the inflammatory cell infiltration in the ileum was not obvious, the villi were normally stretched, and the levels of LPS and Zonulin returned to normal, which were significantly lower than those in the Dox group. The expression of intestinal epithelial tight junction proteins ZO-1, Occludin, and Cludin-1 was significantly increased.

[0064] The specific embodiments of the present invention have been described above. It should be understood that the present invention is not limited to the specific embodiments described above, and those skilled in the art can make various modifications or variations within the scope of the claims, which do not affect the essence of the present invention.

Claims

1. The use of a larazotide polypeptide or a derivative thereof as the sole active ingredient in the preparation of a drug to alleviate the cardiotoxicity of doxorubicin chemotherapy, wherein the larazotide polypeptide has the molecular formula C 32 H 55 N9O 10 The CAS number is 258818-34-7, and the amino acid sequence is: glycylglycyl-L-valine-L-leucyl-L-valine-L-glutamine-L-prolylglycine.

2. The use according to claim 1, characterized in that, Larizolide peptide derivatives include larizolide peptide acetate derivatives.

3. The use according to claim 1, characterized in that, The dosage of larizole polypeptide is as follows: once daily, starting one week before doxorubicin treatment, and once every other day after doxorubicin treatment.

4. The use according to claim 3, characterized in that, The adult dosage of larizole peptide is 0.0385g to 0.154g / day per 70kg body weight.

Images