Application of psoralen as Notch signaling pathway inhibitor

Abstract

The invention relates to an application of psoralen in preparation of a medicine for treating related diseases caused by Notch signaling pathway up-regulation. Drug screening is carried out through an in-vitro cell experiment, and it is found that psoralen can inhibit the activity of a Notch signal channel. The up-regulation of the Notch signal channel is related to the occurrence of various diseases such as prostate cancer, leukemia, breast cancer, cholangiocellular carcinoma, melanoma and glioblastoma, and cell experiments prove that the psoralen can inhibit the survival rate of prostate tumor cell strains. Therefore, the psoralen has the effects of resisting prostate cancer, leukemia, breast cancer, cholangiocellular carcinoma, melanoma, glioblastoma and other diseases. The invention can be applied to medicine and veterinary medicine, the application way comprises injection, slow release technology or compound psoralen in the stent material and other administration ways, and the compound can also be applied together with other supplementary active ingredients, reagents, drugs or hormones.

Description

Technical Field

[0001] The present invention relates to the use of psoralen as a Notch signaling pathway inhibitor in the preparation of a medicine for treating related diseases caused by up-regulation of the Notch pathway, and belongs to the field of biomedicine. Background Art

[0002] The Notch signaling pathway plays a vital role in the initiation and progression of tumors. Notch can play a carcinogenic or local inhibitory role in different cancers. Notch signaling pathway gene mutations or dysregulation can show dual biological characteristics in tumor occurrence and progression. Notch1 mutations can be seen in 50% of acute lymphocytic leukemias, and chronic lymphocytic leukemia, splenic marginal zone B-cell lymphoma, squamous cell lung cancer, and salivary adenoid cystic carcinoma are also associated with Notch gene gain-of-function mutations. In addition, many solid tumors including prostate cancer, breast cancer, melanoma, and lung cancer are associated with abnormal activation of the Notch signaling pathway.

[0003] In addition, Notch signaling can interact with other signals to promote tumorigenesis and progression. Notch signaling is involved in the occurrence of leukemia and breast cancer through its interaction with the NF-kB pathway. Notch signaling can also inhibit apoptosis of cervical cancer cells through the mTOR-Rictor pathway, leading to the occurrence of cervical cancer.

[0004] Given the key role of the Notch signaling pathway in tumors, Notch is a promising therapeutic target. GSIs are the first and most widely studied small molecule Notch inhibitors. Initially, GSIs were developed to treat Alzheimer's disease, and the mechanism used for cancer treatment is to inhibit the cleavage of γ-secretase, which can mediate the generation of NICD. Therefore, GSIs can block the transduction of Notch signals. However, studies have shown that systemic inhibition of Notch signaling by GSIs can cause excessive secretory substances to accumulate in the intestine, resulting in toxic effects "targeted" to the gastrointestinal tract. Therefore, new selective targeted inhibitors for Notch ligands and receptors have become a hot topic of research. In addition, other therapies targeting Notch are currently under study, such as natural products, viral therapies, and so on.

[0005] Currently, there are no research reports on the role of psoralen in inhibiting the activity of Notch signaling pathway and inhibiting the survival rate of tumor cells. Summary of the invention

[0006] In view of the above-mentioned deficiencies in the prior art, one of the objects of the present invention is to provide the use of psoralen as a Notch signaling pathway inhibitor; a second object of the present invention is to provide the use of psoralen in the preparation of a drug for treating related diseases caused by upregulation of the Notch signaling pathway; a third object of the present invention is to provide an anti-tumor drug, in which psoralen is combined with a pharmaceutically acceptable excipient and / or carrier suitable for medical use to prepare a drug for medical and veterinary applications; psoralen can be compounded using various injections, mucosal administration, sustained-release technologies or stent materials.

[0007] The present invention screens drugs through in vitro cell experiments and finds that psoralen can inhibit the activity of the Notch signaling pathway. At present, research in this field has shown that upregulation of the Notch signaling pathway can lead to the occurrence of various diseases such as prostate cancer, leukemia, breast cancer, cholangiocarcinoma, melanoma, glioblastoma, etc. Cell experiments have shown that psoralen can inhibit the survival rate of prostate tumor cell lines. Therefore, psoralen has the effect of resisting diseases such as prostate cancer, leukemia, breast cancer, cholangiocarcinoma, melanoma, glioblastoma, etc.

[0008] The molecular formula of psoralen is C 11 H6O3, molecular structure is shown in formula 1:

[0009]

[0010] To achieve the goal, the technical solution of the present invention is as follows:

[0011] Application of psoralen in the preparation of Notch signaling pathway inhibitors.

[0012] Furthermore, the Notch signaling pathway inhibitor is used in the preparation of drugs for related diseases caused by up-regulation of the Notch signaling pathway.

[0013] Furthermore, the related diseases caused by the upregulation of the Notch signaling pathway include one or more of prostate cancer, acute lymphocytic T-cell leukemia, chronic lymphocytic leukemia, splenic marginal zone lymphoma, diffuse large cell B lymphoma, breast cancer, cholangiocarcinoma, melanoma, glioblastoma, etc.

[0014] Furthermore, psoralen inhibits the activity of Notch signaling pathway.

[0015] Application of psoralen in the preparation of drugs for diseases related to up-regulation of Notch signaling pathway.

[0016] Furthermore, the drug can be prepared into any pharmaceutically acceptable dosage form.

[0017] Furthermore, the dosage form of the drug includes one of injection, liposome, gel preparation, tablet, capsule, granule, powder, oral liquid, pill, spray and the like.

[0018] Furthermore, the drug can be combined with pharmaceutically acceptable excipients and / or carriers suitable for medical use to prepare drugs for medical and veterinary applications.

[0019] Furthermore, the drug intervention methods include various injections, mucosal administration, sustained-release technology or compound drugs in stent materials. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 The results show that different concentrations of psoralen (10 μM, 1 μM, 0.1 μM) in Example 1 intervened in 293T cells and had an inhibitory effect on the activity of the Notch signaling pathway.

[0021] Figure 2 The results show that different concentrations of psoralen (10 μM, 1 μM, 0.1 μM, 0.01 μM) in Example 3 intervened in DU145 cells and had an inhibitory effect on cell survival rate. DETAILED DESCRIPTION

[0022] The present invention will be further described below in conjunction with the accompanying drawings and specific embodiments. These embodiments should be understood to be only used to illustrate the present invention and not to limit the scope of protection of the present invention. After reading the contents of the present invention, those skilled in the art may make various changes or modifications to the present invention, and these equivalent changes and modifications also fall within the scope defined by the claims of the present invention.

[0023] Example 1 - Study on the inhibition of Notch signaling pathway activity by psoralen

[0024] 1. Cell line: 293T cell line.

[0025] 2. Cell passaging and plating: When the cells grow to a confluence of 90%, they are passaged and plated in 24-well plates, with 50,000 cells per well.

[0026] 3. Drug intervention: When the cells grew to a confluence of 70-80%, 0, 0.1 μM, 1 μM, and 10 μM psoralen were used to intervene in 293T cells.

[0027] 4. Notch signaling pathway reporter gene transfection: 2 hours after drug intervention, the Notch signaling pathway reporter gene was transfected with Lipo3000 reagent (L3000008, Invitrogen TM ) The Notch signaling pathway reporter gene plasmid (RBP-Jk Luc luciferase reporter gene plasmid, Shanghai Yisheng) was transfected into 293T cells, and a negative control group was set up.

[0028] 5. Index detection: After 24 hours, the fluorescence intensity of each group of cells was detected using a dual luciferase reporter gene detection kit (Shanghai Yisheng).

[0029] 6. Experimental results: Figure 1 As shown in the figure, compared with the negative control group (Ctrl group), the fluorescence intensity of the cells increased significantly after transfection of the RBP-Jk Luc luciferase reporter gene plasmid (##, P<0.01), indicating that the endogenous Notch signaling pathway was activated in the cells. After intervention with 0.1μM, 1μM, and 10μM psoralen for 24 hours, the fluorescence intensity of each group decreased significantly compared with the RBP-Jk group (*, P<0.05; **, P<0.01), indicating that the activity of the endogenous Notch signaling pathway was significantly inhibited.

[0030] Example 2 - Study on the inhibition of prostate tumor cell survival rate by psoralen

[0031] 1. Cell line: Prostate cancer cell line DU145.

[0032] 2. Cell passaging and plating: When the cells grow to a confluence of 90%, they are passaged and plated in 96-well plates.

[0033] 3. Drug intervention: When the cells grew to a confluence of 50%, DU145 cells were intervened with 0, 0.01 μM, 0.1 μM, 1 μM, and 10 μM psoralen for 72 hours.

[0034] 4. Index detection: After discarding the culture medium, add complete culture medium containing CCK-8 reagent and continue culturing for 1 hour. Use an enzyme reader to detect the absorbance at 450nm and calculate the cell survival rate.

[0035] 5. Experimental results: Figure 2 As shown, compared with the control group (Ctrl group) and the solvent group (DMSO group), the survival rate of DU145 cells in each concentration of psoralen groups was significantly reduced, and the inhibitory effect became more significant with the decrease of the dose (compared with the Ctrl group, ##, P<0.01; compared with the DMSO group, **, P<0.01), indicating that psoralen can inhibit the survival rate of tumor cells.

Claims

1. Use of psoralen as an inhibitor of Notch signaling pathway.

2. The application of psoralen in the preparation of drugs for treating related diseases caused by upregulation of Notch signaling pathway.

3. The use according to claim 2, characterized in that: Diseases related to upregulation of the Notch signaling pathway include one or more of prostate cancer, acute lymphocytic T-cell leukemia, chronic lymphocytic leukemia, splenic marginal zone lymphoma, diffuse large cell B-cell lymphoma, breast cancer, cholangiocarcinoma, melanoma, glioblastoma, etc.

4. The use according to any one of claims 1 to 3, characterized in that: Psoralen exerts its therapeutic effect by inhibiting the activity of the Notch signaling pathway and suppressing the survival rate of tumor cells.

5. The use according to any one of claims 1 to 3, characterized in that: Psoralen is combined with pharmaceutically acceptable excipients and / or carriers suitable for pharmaceutical use.

6. The use according to any one of claims 1 to 3, characterized in that: Psoralen can be prepared into any pharmaceutically acceptable dosage form.

7. The use according to any one of claims 1 to 3, characterized in that: Psoralen intervention methods include various injections, mucosal administration, sustained-release technology, or psoralen complexed in stent materials.

Images