Application of pyropheophorbide-alpha in preparation of medicine for treating chronic pancreatitis

Abstract

The invention relates to the technical field of traditional Chinese medicines, in particular to application of a small molecular compound pyropheophorbide-alpha screened from a marine natural product library in preparation of a medicine for treating chronic pancreatitis. The application has the advantages that in-vivo and in-vitro fibrosis resistance and chronic pancreatitis treatment efficacy of pyropheophorbide-alpha are found, and the pyropheophorbide-alpha has a wide prospect in clinical transformation and is expected to bring good news to CP patients.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, specifically to the application of the marine natural product pyrophyllite-alpha in the preparation of a drug for treating chronic pancreatitis. Background Technology

[0002] Chronic pancreatitis (CP) is a progressive fibrotic disease of the pancreas. Clinical symptoms primarily include abdominal pain, steatorrhea, and other manifestations of pancreatic exocrine and endocrine dysfunction, severely impacting patients' daily lives. The pathological features of CP are complex, including pancreatic parenchymal atrophy, pancreatic duct stones, fibrosis caused by extracellular matrix deposition, and infiltration of inflammatory cells such as macrophages and lymphocytes. Currently, the global prevalence of CP is 92 per 100,000, and it is increasing annually.

[0003] For a long time, the treatment of pancreatic cystic inflammatory disease (CP) has focused on controlling symptoms and preventing complications. Current clinical treatments for CP include pain relief, pancreatic enzyme replacement therapy, endoscopic sphincterotomy, stent placement, extracorporeal shock wave lithotripsy, and surgery. While these methods can effectively alleviate symptoms, they cannot slow the progression of fibrosis in CP, thus failing to fundamentally cure the disease. The continued progression of inflammation and fibrosis not only leads to the loss of pancreatic endocrine and exocrine function but also increases the risk of pancreatic cancer by 15 times compared to the general population. Therefore, finding drugs that can slow disease progression and improve the quality of life and prognosis of CP patients has become an urgent need in CP treatment.

[0004] Marine natural products provide crucial compound information for modern innovative drug development. Currently, over 35,000 marine natural products have been discovered. Due to the unique marine environment, these products possess novel structures and unique physiological functions, with more than half exhibiting biological activity. They are a vital source of lead compounds for drugs treating major human diseases such as cancer, cardiovascular and cerebrovascular diseases, immune diseases, and nervous system diseases. However, to date, only 13 new marine drugs have been marketed. The marine biopharmaceutical industry has enormous potential, and exploring marine resources for drug discovery and strengthening the screening of marine active drugs will be an essential path for future new drug development. Summary of the Invention

[0005] The purpose of this invention is to screen a small molecule compound, pyropheophorbide-a, from a marine natural product library that can inhibit pancreatic fibrosis and treat CP. Pyropheophorbide-a can alleviate pancreatic fibrosis in CP and delay the progression of CP.

[0006] A first aspect of the present invention provides the use of pyrophyllite-alpha in the preparation of a medicament for treating chronic pancreatitis.

[0007] The pyrophyllophosphochlorophyll-alpha mentioned above is a small molecule compound screened from a marine natural product library, and its structural formula is shown in Formula I below:

[0008]

[0009] Furthermore, the medication for treating chronic pancreatitis is a drug that alleviates pancreatic fibrosis and slows the progression of chronic pancreatitis.

[0010] Furthermore, the drug comprises:

[0011] (A) an effective amount of pyrophylloxera-alpha; and

[0012] (B) Pharmaceutically or immunologically acceptable carriers or excipients.

[0013] Furthermore, in the aforementioned applications, bamboo red fungus extract is used in the preparation of drugs that improve pancreatic weight loss, reduce the degree of fibrosis, and decrease the expression level of fibrosis-related fibronectin in pancreatic tissue.

[0014] Furthermore, in the aforementioned applications, bamboo red fungus extract is used in the preparation of drugs that inhibit the synthesis of HPSC fibrosis-related proteins.

[0015] Furthermore, in the aforementioned drug, the dosage of bamboo red fungus A is 1.25 mg / kg.

[0016] A second aspect of the invention provides the use of pyrophyllate-alpha in the preparation of a medicament for inhibiting pancreatic fibrosis.

[0017] Experiments of this invention demonstrate that:

[0018] Pyrophyllofoate-alpha improved the decrease in pancreatic weight induced by taeniacin. Figure 1 Further investigation using HE and Masson staining revealed pathological changes in pancreatic tissue. After treatment with pyrophylloxera-alpha, acinar cell atrophy in CP mice was alleviated, the parenchymal tissue remained relatively intact, and the degree of fibrosis was reduced. Figure 2 To investigate the changes in fibrosis-related protein content in pancreatic tissue after treatment with pyrophyllin-alpha, proteins were extracted from pancreatic tissue, and the expression level of fibrosis-related fibronectin (FN) was detected by Western blot. The results showed that pyrophyllin-alpha reduced the expression level of FN in the pancreatic tissue of CP mice. Figure 3 This confirmed the anti-fibrotic effect of the drug. TGF-β1 stimulation significantly increased the expression of fibrosis-related proteins, while pyrophyllin-alpha dose-dependently decreased the content of HPSC fibrosis-related proteins. Figure 4 ).

[0019] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0020] This invention has discovered the in vitro and in vivo anti-fibrotic and therapeutic effects of pyrophyllofoetin-alpha, a small molecule compound screened from a marine natural product library, on chronic pancreatitis. It has broad prospects for clinical translation and is expected to bring good news to CP patients. Attached Figure Description

[0021] Figure 1 The ratio of mouse pancreatic tissue to body weight (mg / g). Groups: control: control group; CP: chronic pancreatitis model group; CP+Pyropheophorbide-a: chronic pancreatitis pyropheophorbide-alpha treatment group.

[0022] Figure 2 HE staining and Masson staining of mouse pancreatic tissue. Grouping was the same. Figure 1 .

[0023] Figure 3 Western blot analysis of FN expression levels in mouse pancreatic tissue. Groups were the same. Figure 1 .

[0024] Figure 4 Results of Western blot experiments using HPSC. COL3a1: Type III collagen α1 chain; Fibronectin: fibronectin; GAPDH: internal reference protein. Detailed Implementation

[0025] The specific implementation methods provided by the present invention will be described in detail below with reference to the embodiments.

[0026] Example 1:

[0027] 1. Animal experiments:

[0028] Six-week-old C57BL / 6J mice were randomly divided into three groups: a control group, a CP group, and a CP + pyrophyllin-alpha group. The CP group was induced by intraperitoneal injection of phytohemagglutinin at a dose of 50 μg / kg three times a week for four weeks. Pyrophyllin-alpha was administered via gavage after dissolving in corn oil, once daily for four weeks at a dose of 1.25 mg / kg. The control and CP groups received the same volume of corn oil via gavage. Forty-eight hours after induced fibrosis, the mice were dissected, and pancreatic tissue was weighed. The pancreatic head was paraffin-embedded and stained with hematoxylin and eosin (HE) and Masson's stain. The remaining pancreatic tissue was used for Western blot detection of fibrosis-related proteins.

[0029] 2. Cell experiments:

[0030] The anti-fibrotic effect of pyrophyllin-alpha was verified using human pancreatic stellate cells (HPSCs). PSCs were stimulated with TGF-β1 (10 ng / mL) to simulate the activation state of HPSCs during CP fibrosis. Groups were divided into: control group, TGF-β1 modeling group, and TGF-β1 modeling + pyrophyllin-alpha treatment group, with drug concentrations of 0, 0.5, 1, 2, and 4 μM in the treatment groups, respectively. The control and treatment groups were treated with the same volume of DMSO. Samples were collected and proteins extracted after 24 hours. Changes in fibrosis-related protein content were analyzed using Western blot experiments.

[0031] 3. Results and Analysis

[0032] (1) Pyromethes-phosphochlorophyll-alpha alleviates pancreatic tissue fibrosis in CP mice

[0033] To investigate the anti-fibrotic effect of pyrophyllin-alpha in a CP mouse model, the pancreas of mice was dissected 4 weeks after modeling. The results showed that pyrophyllin-alpha improved the decrease in pancreatic weight induced by taurine. Figure 1 ).

[0034] Further observation of pancreatic tissue pathological changes using HE and Masson staining revealed extensive inflammatory cell infiltration in the CP model group, predominantly lymphocytes and plasma cells, indicating pancreatic structural destruction, acinar cell atrophy, and significant fibrosis around the acini and between lobules, with a marked increase in interlobular spacing. After treatment with pyrophyllofoetin-alpha, acinar cell atrophy was alleviated, the parenchymal tissue remained relatively intact, and the degree of fibrosis decreased. Figure 2 ).

[0035] To investigate the changes in fibrosis-related protein content in pancreatic tissue after treatment with pyrophyllin-alpha, proteins were extracted from pancreatic tissue, and the expression level of fibrosis-related fibronectin (FN) was detected by Western blot. The results showed that pyrophyllin-alpha reduced the expression level of FN in the pancreatic tissue of CP mice. Figure 3 This confirmed the drug's anti-fibrotic effect.

[0036] (2) Pyrophyllofop-alpha inhibits the synthesis of HPSC fibrosis-related proteins in vitro.

[0037] Activation of the hypothalamic-pituitary-sac (HPSC) and the secretion of fibrosis-related proteins are key to the development of pancreatic fibrosis in CP (proliferative pancreatic disease). Therefore, we investigated the effect of pyrophyllin-alpha on HPSC in vitro. PSCs were stimulated for 24 hours with the common HPSC activator TGF-β1 (10 ng / mL), and simultaneously treated with different concentrations of pyrophyllin-alpha (0, 0.5, 1, 2, 4 μM). The results showed that TGF-β1 stimulation significantly increased the expression of fibrosis-related proteins, while pyrophyllin-alpha dose-dependently decreased the content of HPSC fibrosis-related proteins. Figure 4 ).

[0038] The preferred embodiments of the present invention have been described in detail above, but the present invention is not limited to the embodiments described. Those skilled in the art can make various equivalent modifications or substitutions without departing from the spirit of the present invention, and these equivalent modifications or substitutions are all included within the scope defined by the claims of this application.

Claims

1. Application of pyrophyllite-alpha in the preparation of drugs for treating chronic pancreatitis.

2. The application of pyrophyllite-alpha according to claim 1 in the preparation of a medicament for treating chronic pancreatitis, characterized in that, The medication mentioned for treating chronic pancreatitis is designed to alleviate pancreatic fibrosis and slow the progression of chronic pancreatitis.

3. The application of pyrophyllite-alpha according to claim 1 in the preparation of a medicament for treating chronic pancreatitis, characterized in that, The drug comprises: (A) an effective amount of pyrophylloxera-alpha; and (B) Pharmaceutically or immunologically acceptable carriers or excipients.

4. The application of pyrophyllite-alpha according to claim 1 in the preparation of a medicament for treating chronic pancreatitis, characterized in that, Application of bamboo red fungus A in the preparation of drugs that improve pancreatic weight loss, reduce fibrosis, and decrease the expression of fibrosis-related fibronectin in pancreatic tissue.

5. The application of pyrophyllite-alpha according to claim 1 in the preparation of a medicament for treating chronic pancreatitis, characterized in that, Application of bamboo red fungus A in the preparation of drugs that inhibit the synthesis of HPSC fibrosis-related proteins.

6. The use of pyrophyllite-alpha according to claim 1 in the preparation of a medicament for treating chronic pancreatitis, characterized in that, In the aforementioned drug, the dosage of bamboo red fungus A is 1.25 mg / kg.

7. Application of pyrophyllite-alpha in the preparation of drugs that inhibit pancreatic fibrosis.

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