Ntsr1 receptor antagonists or drugs and uses thereof

Abstract

This invention relates to the field of pharmaceutical technology, and discovers that the natural products boldinine and norboldinine can be used as NTSR1 (Neurotensin receptor-1, NTSR1) receptor antagonists and their applications. The compounds are boldinine and norboldinine ((+)-Laurolitsine), which have NTSR1 receptor antagonistic activity and are NTSR1 receptor antagonists that can prevent and treat alcohol poisoning, addiction, schizophrenia, autism spectrum disorder, and mood disorders. Accordingly, this invention can provide novel NTSR1 receptor antagonist prospective compounds with clear targets for the above-mentioned related diseases.

Description

Technical Field

[0001] This invention belongs to the field of NTSR1 (Neurotensin receptor-1) receptor agonists, and relates to the discovery of targets for alkaloid compounds, specifically the targets of boridine and norboridine. The alkaloid compounds boridine and norboridine, and their derivatives, as well as their corresponding pharmaceutically acceptable salt-forming compounds; the target is the NTSR1 receptor; the applications are for alcoholism, addiction, schizophrenia, autism spectrum disorders, and mood disorders. Background Technology

[0002] Neurotensin receptor-1 (NTSR1) is a family of G protein-coupled receptors (GPCRs), a typical class A GPCR. It mediates and regulates neurotransmission through endogenous neurotensin (NTS), and is widely involved in regulating blood pressure, blood glucose, body temperature, analgesia, and neuronal damage. NTSR1 is primarily recognized and regulated by GRK2 and GRK5. Studies have reported that NTSR1 is an important drug target for the treatment of addiction, autism spectrum disorders, alcoholism, obesity, analgesia, cancer, Parkinson's disease, and schizophrenia. Therefore, the discovery and confirmation of new ligands for the NTSR1 receptor is of great significance for elucidating its pharmacological and biological functions, which will provide new directions for targeted therapy of the disease (Polak JM, Bloom S R. Ann Ny Acad Sci, 1982, 400: 75-93; Carraway R, Leeman S E. Journal of Biological Chemistry, 1973, 248(19): 6854-6861; Vincent JP, Mazella J, Kitabgi P. Trends in pharmacological sciences, 1999, 20(7): 302-309; Popp E, Schneider A, Vogel P, et al. Neuropeptides, 2007, 41(5): 349-354; St-Gelais F, Jomphe C, Trudeau L EJ Psychiatr Neurosci, 2006, 31(4): 229-245; Mustain WC, Rychahou PG, Evers B M. Curr Opin Endocrinol, 2011, 18(1): 75-82; Carraway RE, Plona A M. Peptides, 2006, 27(10): 2445-2460).

[0003] Currently, there are no reports on the effects of alkaloid compounds bortine and norbortine on diseases such as alcoholism, addiction, schizophrenia, autism spectrum disorder, and mood disorders based on the NTSR1 (Neurotensin receptor-1) receptor. Summary of the Invention

[0004] This invention relates to the discovery of the target of bordin and norbordin in alkaloid compounds and the application of such compounds. One objective is to provide information that the target of bordin and norbordin is the NTSR1 receptor. Another objective is to provide information on the clinical application of such compounds in diseases such as alcoholism, addiction, schizophrenia, autism spectrum disorder, and mood disorders.

[0005] The technical solution of this invention is as follows:

[0006] The NTSR1 receptor antagonists are the compounds Boldine and (+)-Laurolitsine.

[0007] The chemical structures of bortine base and norbortine are as follows:

[0008]

[0009] The present invention relates to the application of alkaloid compounds in the preparation of drugs for the prevention and / or treatment of diseases such as alcohol poisoning, addiction, schizophrenia, autism spectrum disorder, and mood disorders, wherein the alkaloid compounds are boridine base and norboridine.

[0010] The beneficial effects of this invention are:

[0011] In vitro cell experiments have shown that the compounds in this invention, bortine and norbortine, act on the NTSR1 receptor, which is a G protein-coupled receptor. The NTSR1 receptor is associated with diseases such as alcoholism, addiction, schizophrenia, autism spectrum disorder, and mood disorders. Based on the correlation between the target and the disease, the clinical application scope of such compounds can be broadened.

[0012] This invention discovers that the natural products bordinine and norbordin can serve as NTSR1 (Neurotensin receptor-1) receptor antagonists and their applications. Specifically, it involves the discovery of the target of alkaloid compounds, namely the NTSR1 receptor. The NTSR1 receptor antagonist is bordinine and norbordin or their derivatives, and one or more of their corresponding pharmaceutically acceptable salts as active ingredients. In vitro cell experiments demonstrate that the compounds of this invention are NTSR1 receptor antagonists. Current research indicates that the NTSR1 receptor is associated with diseases such as alcoholism, addiction, schizophrenia, autism spectrum disorder, and mood disorders. NTSR1 receptor antagonists can prevent and treat these diseases, thus providing novel NTSR1 receptor antagonist prospective compounds with clearly defined targets for the aforementioned related diseases. Attached Figure Description

[0013] Figure 1 The negative control group (HBSS buffer containing 0.1% DMSO and 20 mM HEPES), neurotensin (2 nM), bordrine (20 μM), and norbordrine (20 μM), as well as the maximum DMR response (%) of bordrine-neurotensin and norbordrine-neurotensin in HT-29 cells within 60 min;

[0014] Figure 2 The maximum DMR response-response curves of different concentrations of bortine (A) and norbortine (B) in HT-29 cells over 60 min for activation and desensitization are shown. Detailed Implementation

[0015] The present invention will now be further illustrated with examples. These examples are for illustrative purposes only and are not intended to limit the scope of the invention.

[0016] Neurotensin (brand: Yuanye Biotechnology; catalog number: S85240), bortine (brand: Yuanye Biotechnology; catalog number: B30030), and norbortine (brand: Yuanye Biotechnology; catalog number: B50112) were purchased from Shanghai Yuanye Biotechnology Co., Ltd. HT-29 cells (catalog number: TCHU103) were purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences. McCoy's 5A medium (brand: VivaCell; catalog number: C3020-0500) was purchased from Shenyang Huibai Biotechnology Co., Ltd. Fetal bovine serum (FBS) (brand: PAN; catalog number: ST30-3302) was purchased from Suzhou Qianshe Biotechnology Co., Ltd.; dimethyl sulfoxide (DMSO, catalog number: A503039-0250) was also purchased. HBSS (brand: Gibco; catalog number: 14065056; specification: (10×) 500mL) and HEPES (brand: Gibco; catalog number: 15630080; specification: (100×) 500mL) buffer solutions were purchased from Invitrogen (Shanghai) Trading Co., Ltd. Each compound was dissolved in DMSO to prepare a 100mM stock solution. The buffer solution was prepared at a volume ratio of HEPES:HBSS:ultrapure water = 1:5:44, and was used for compound dilution during the experiment. The detection platform was Corning Gorilla Glass 3. The imager detects the wavelength shift caused by dynamic mass resetting (DMR) of cells.

[0017] Example 1: Screening of NTSR1 receptor activity of compounds

[0018] HT-29 cells in the logarithmic growth phase were seeded into... In the 384-well biosensor microplate, each well was seeded with 40 μL of cell suspension (McCoy's 5A medium containing cells) and 3.0 × 10⁶ cells per well.4 Then, place the 384-well plate in a cell culture incubator (air with a volume concentration of 5% CO2, 37°C) and culture for 22-24 hours. When the cell confluence reaches about 95%, conduct the experiment.

[0019] First, 10 μL of 20 μM bortine, 10 μL of 20 μM norbortine, and 10 μL of buffer (negative control, HBSS buffer containing 0.1% DMSO (v / v) and 20 mM HEPES) were added to different wells seeded with HT-29 cells, and the cells were analyzed on an EPIC instrument for 1 hour. Then, 2 nM of neurotensin (based on the final volume per well) was added, and the cells were analyzed on an EPIC instrument for another hour. The results are as follows. Figure 1 As shown, bortine and norbortine showed virtually no DMR response signal in HT-29 cells, but they could inhibit the DMR response signal of the NTSR1 receptor agonist neurotensin, with inhibition rates of 64.4% and 52.1%, respectively, indicating that they may have NTSR1 receptor antagonistic activity.

[0020] Example 2: Verification of the NTSR1 receptor antagonistic activity of the compound

[0021] HT-29 cells in the logarithmic growth phase were seeded into... In the 384-well biosensor microplate, each well was seeded with 40 μL of cell suspension (McCoy's 5A medium containing cells) and 3.0 × 10⁶ cells per well. 4 Then, place the 384-well plate in a cell culture incubator (air with a volume concentration of 5% CO2, 37°C) and culture for 22-24 hours. When the cell confluence reaches about 95%, conduct the experiment.

[0022] DMR antagonism assay: Bordeaux mixture and norbordine at different final concentrations (100 μM, 33.33 μM, 11.11 μM, 3.70 μM, 1.23 μM, 0.41 μM, 0.14 μM, containing 0.1% DMSO (volume ratio)) were pre-added to different wells inoculated with HT-29 cells, and their DMR response signals were monitored in real-time on an EPIC instrument for 1 h. Then, the NTSR1 receptor agonist neurotensin (2 nM, calculated based on the final volume per well) was added, and the monitoring was performed in real-time on an EPIC instrument for 1 h. The results are as follows: Figure 2 As shown, bortine and norbortine showed virtually no DMR response signal in HT-29 cells, but inhibited the DMR response signal of the NTSR1 receptor agonist neurotensin in a concentration-dependent manner, indicating that they possess NTSR1 receptor antagonistic activity and are NTSR1 receptor antagonists. Their antagonistic IC50 value is [not specified in the original text]. 50 The values ​​were 5.818±0.931 μM and 38.930±7.835 μM.

[0023] Pharmacological DMR experiments have shown that the NTSR1 receptor is the target of the compounds bortine and norbortine. Current research indicates that the NTSR1 receptor is associated with diseases such as alcoholism, addiction, schizophrenia, autism spectrum disorder, and mood disorders. Based on the correlation between the target and these diseases, the clinical applications of these compounds can be broadened.

Claims

1. The use of boridine base and / or norboridine compounds as NTSR1 receptor antagonists or in the preparation of drugs for NTSR1 receptor-related diseases.

2. The application according to claim 1, characterized in that: The boridine base and norboridine compound include one or more of the following: boridine base, norboridine, boridine base derivatives, norboridine derivatives, or pharmaceutically acceptable salts of boridine base and pharmaceutically acceptable salts of norboridine. The NTSR1 receptor antagonist or the prepared drug contains one or more of the following as active ingredients: boridine base, norboridine, boridine base derivatives, norboridine derivatives, or pharmaceutically acceptable salts of boridine base and norboridine.

3. The application according to claim 1 or 2, characterized in that: The structural formulas of the compounds boldinine and norboldinine ((+)-Laurolitsine) are as follows:

4. The application according to claim 1, characterized in that: The prepared drug is a drug for the treatment and / or prevention of one or more diseases including alcoholism, addiction, schizophrenia, autism spectrum disorder, and mood disorders.

5. The application according to claim 1, 2 or 4, characterized in that: The NTSR1 receptor antagonist or drug also includes a pharmaceutically acceptable carrier and / or excipient.

6. A drug or NTSR1 receptor antagonist, characterized in that: The active ingredient is one or more of the following: boridine base, norboridine, boridine base derivatives, norboridine derivatives, pharmaceutically acceptable salts of boridine base, and pharmaceutically acceptable salts of norboridine.

7. The drug or NTSR1 receptor antagonist according to claim 6, characterized in that: The chemical structures of boldinine and (+)-laurolitsine are shown below:

8. The drug according to claim 6 or 7, characterized in that: The drug or NTSR1 receptor antagonist also includes a pharmaceutically acceptable carrier and / or excipient.

9. The drug or NTSR1 receptor antagonist according to claim 6 or 7, characterized in that: The drug is for the treatment and / or prevention of one or more of the following diseases: alcoholism, addiction, schizophrenia, autism spectrum disorder, and mood disorders.

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