Icaritin derivatives and their use in the treatment of liver cancer

Abstract

The application discloses an icariin derivative in the technical field of medicines, characterized by a structural formula as shown in formula (1), and the icariin derivative of formula (1) has good anti-liver cancer activity, high efficiency and low toxicity, and can be developed into a new medicine for treating liver cancer.

Description

Technical Field

[0001] This invention relates to the field of biomedicine, specifically to icariin derivatives and their application in the treatment of liver cancer. Background Technology

[0002] Hepatocellular carcinoma (HCC) is one of the most common malignant tumors worldwide, characterized by high malignancy and a high rate of recurrence and metastasis. Hepatectomy remains one of the preferred treatments for HCC. However, hepatectomy usually cannot prevent tumor metastasis, carries high surgical risks, is prone to postoperative complications, and has a poor prognosis. The 5-year recurrence rate after surgery is 40%–70%, significantly impacting long-term survival for patients.

[0003] Postoperative recurrence of liver cancer includes two modes: intrahepatic metastasis and multicentric occurrence. The former is due to the presence of micrometastases in the residual liver and usually occurs within 2 years (early recurrence). The latter is due to underlying liver disease or new tumors developing on the basis of cirrhosis and usually occurs after 2 years (late recurrence). Studies have shown that patients with late recurrence have better outcomes with surgical treatment compared to those with early recurrence. Currently, surgical treatments for recurrent liver cancer include re-hepatectomy, salvage liver transplantation, and radiofrequency ablation, each with its own advantages and disadvantages. Re-hepatectomy is one of the preferred treatments for recurrent liver cancer, but its application is limited by varying degrees of cirrhosis, insufficient residual liver volume, and abdominal adhesions. Salvage liver transplantation is also a relatively effective treatment option for recurrent liver cancer, but donor livers are scarce. Radiofrequency ablation has advantages such as repeatability and minimally invasiveness, but its local recurrence rate is high. Therefore, choosing the most appropriate surgical treatment plan for patients with recurrent liver cancer remains a controversial topic. The selection of a more appropriate surgical treatment plan needs to be guided by the tumor condition at the time of the initial hepatectomy, the characteristics of the recurrent tumor, the patient's basic condition, and the time of recurrence. Currently, there is still a lack of effective treatments to radically cure liver cancer.

[0004] Chemotherapy is a commonly used clinical treatment, but it uses cytotoxic drugs that not only adversely affect healthy organs but also cause serious side effects such as bleeding and anemia. Furthermore, these side effects increase the mortality rate of cancer patients. Therefore, the development of highly effective and low-toxicity drugs is urgently needed.

[0005] Epimedium is a traditional Chinese medicine, belonging to the genus Epimedium in the Berberidaceae family. It has a pungent and sweet taste and is warm in nature. Its dried leaves are used medicinally, primarily to treat impotence, seminal emission, kidney yang deficiency, rheumatic pain, weakness of muscles and bones, numbness, and contractures. Modern pharmacological studies have shown that the main active components of Epimedium extract include icariin, icariin derivatives, total flavonoid derivatives of Epimedium, as well as alkaloids, anthraquinones, lignans, phytosterols, anthocyanins, terpenoids, chlorogenic acid, essential fatty acids, trace elements, and other bioactive and nutrient components. It not only possesses functions such as dilating coronary arteries, inhibiting microorganisms, anti-aging, and promoting bone cell growth, but also exhibits various biological activities such as inhibiting tumors.

[0006] Icaritin (ICT) is the active flavonoid component of Epimedium, and its structure is shown below. It is formed by the hydrolysis of icariin, the main active monomer of Epimedium. Studies have shown that icariin has multiple effects, including anti-tumor, anti-liver fibrosis, anti-osteoporosis, prostate function improvement, neuroprotection, immunosuppression, anti-inflammation, antioxidant, and estrogen receptor-like effects. Furthermore, it has a broad spectrum of anti-tumor activity, inhibiting various tumors.

[0007]

[0008] However, like many natural products, icariin has the disadvantages of poor water solubility and low bioavailability, which greatly limits its clinical application.

[0009] Numerous clinical studies have demonstrated that carbohydrates have many binding receptors on cell surfaces, exhibiting cell-targeting properties and good water solubility, making them excellent carriers for lead drugs. Glycosyl compounds can effectively improve the low bioavailability, poor solubility, poor pharmacological activity, and reduce toxic side effects of certain drugs. Therefore, designing a glycosylated icariin derivative could help improve the bioavailability of icariin and increase its clinical application rate. Summary of the Invention

[0010] To address the shortcomings of existing technologies, this invention designs a glycosylated derivative of icariin.

[0011] One objective of this invention is to provide an icariin derivative, the structural formula of which is shown in formula (1).

[0012]

[0013] The second objective of this invention is to provide the application of the epimedium derivative of formula (1) in the preparation of anti-liver cancer drugs.

[0014] Furthermore, the concentration at which the compound of formula (1) exerts its effect on liver cancer cells is 2–50 μM.

[0015] Furthermore, the concentration at which the compound of formula (1) exerts its effect on liver cancer cells is 10–20 μM.

[0016] Furthermore, the concentration at which the compound of formula (1) exerts its effect on liver cancer cells is 20 μM.

[0017] Furthermore, the reaction time of the compound of formula (1) is 48h.

[0018] The working principle and beneficial effects of the present invention are as follows: 1. Through screening by the MTT method, it was found that the compound of formula (1) has a better inhibitory effect on the growth of human liver cancer cells HepG2 than icariin.

[0019] 2. Screening by the MTT assay revealed that compound (1) was less toxic to normal human liver cells L02 than icariin.

[0020] 3. Screening by CCK-8 method revealed that compound (1) had a better inhibitory effect on the growth of mouse-derived hepatocellular carcinoma cells H22 than icariin.

[0021] Through in vitro antitumor activity screening, it was found that the icariin derivative of formula (1) not only has a good inhibitory effect on liver cancer cells, but also has low toxicity to normal liver cells, avoiding adverse effects on healthy organs during treatment. It has the characteristics of high efficiency and low toxicity and is suitable for clinical application. Attached Figure Description

[0022] Figure 1 A schematic diagram illustrating the effect of icariin derivatives on the viability of HepG2 human liver cancer cells as determined by the MTT assay.

[0023] Figure 2 Schematic diagram of the effect of icariin derivatives on the viability of L02 normal hepatocytes as determined by the MTT assay;

[0024] Figure 3 This is a schematic diagram illustrating the effect of icariin derivatives on the viability of H22 hepatocellular carcinoma cells as determined by the CCK-8 assay. Detailed Implementation

[0025] The following detailed description illustrates the specific implementation method:

[0026] In the following examples, unless otherwise stated, the test methods are generally carried out under conventional conditions or conditions recommended by the manufacturer; the raw materials and reagents shown are all commercially available.

[0027] Example 1. Effect of icariin derivatives on the viability of HepG2 human hepatocellular carcinoma cells using the MTT assay.

[0028] Log-growing human hepatocellular carcinoma cells (HepG2) were added to 96-well plates, with approximately 3000 cells per well (100 μL). The aforementioned icariin derivative was then added, along with icariin as a control. Five parallel wells were prepared for each group, and the plates were incubated at 37°C for 48 hours in a CO2 incubator. Four hours before the end of the experiment, 20 μL / well of MTT (20 mg / mL) solution was added, and the plates were incubated for another 4 hours. The culture medium was then discarded, and 150 μL / well of DMSO was added. After the crystals dissolved, the OD value of each well was measured at 490 nm using a microplate reader. The results are as follows: Figure 1 As shown. From Figure 1 As can be seen, at a drug concentration of 20 μM, the icariin derivative exhibited a stronger inhibitory effect on the growth of human hepatocellular carcinoma cells HepG2 compared to the control drug icariin, and the inhibitory effect increased with increasing drug concentration. Note: * indicates p < 0.05 (data = mean + / - SD)

[0029] Example 2. MTT assay for the effect of icariin derivatives on the viability of L02 human normal hepatocytes

[0030] Log-growing human normal hepatocytes (L02) were added to 96-well plates, with 100 μL per well containing approximately 3000 cells. The aforementioned icariin derivative was then added, along with icariin as a control. Five parallel wells were prepared for each group, and the plates were incubated at 37°C for 48 hours in a CO2 incubator. Four hours before the end of the experiment, 20 μL / well of MTT (20 mg / mL) solution was added, and the plates were incubated for another 4 hours. The culture medium was then discarded, and 150 μL / well of DMSO was added. After the crystals dissolved, the OD value of each well was measured at 490 nm using a microplate reader. The results are as follows: Figure 2 As shown. From Figure 2 As can be seen, within the effective concentration range, the icariin derivative showed no toxicity to normal human L02 hepatocytes compared to the control drug icariin. Note: * indicates p < 0.05 (data = mean + / - SD)

[0031] Example 3. CCK-8 assay to detect the effect of icariin derivatives on the viability of H22 mouse hepatocellular carcinoma cells.

[0032] Logarithmically growing mouse hepatocellular carcinoma cells (H22) were added to 96-well plates, with approximately 3000 cells per well (100 μL). The aforementioned icariin derivative was then added, along with icariin as a control. Five parallel wells were prepared for each group, and the plates were incubated at 37°C for 48 hours in a CO2 incubator. Four hours before the end of the experiment, 20 μL / well of CCK-8 solution was added, and the plates were incubated for another 4 hours. The OD value of each well was measured at 450 nm using a microplate reader. The results are as follows: Figure 3 As shown. From Figure 3As can be seen, the icariin derivative has a stronger inhibitory effect on the growth of H22 cells than the control drug icariin, and the inhibitory effect increases with increasing drug concentration. Note: * indicates p < 0.05 (data = mean + / - SD).

[0033] The above descriptions are merely embodiments of the present invention, and common knowledge regarding specific structures and characteristics is not elaborated upon here. It should be noted that those skilled in the art can make various modifications and improvements without departing from the structure of the present invention, and these should also be considered within the scope of protection of the present invention. These modifications and improvements will not affect the effectiveness of the present invention or the practicality of the patent. The scope of protection claimed in this application should be determined by the content of its claims, and the specific embodiments described in the specification can be used to interpret the content of the claims.

Claims

1. A derivative of icariin, characterized in that, The structural formula is shown as formula (1), 2. The use of the icariin derivative according to claim 1 in the preparation of a liver cancer resisting drug.

3. Use according to claim 2, characterized in that: The concentration of the compound of formula (1) acting on the liver cancer cells is 2-50 μM.

4. Use according to claim 3, characterized in that: The concentration of the compound of formula (1) acting on the liver cancer cells is 10-20 μM.

5. Use according to claim 4, characterized in that: The concentration of the compound of formula (1) acting on the liver cancer cells is 20 μM.

6. Use according to claim 5, characterized in that: The acting time of the compound of formula (1) is 48 h.

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