Method for preparing compound and use thereof as olfactory receptor OLFR110 agonist

Abstract

Provided in the present invention are a method for preparing a compound and the use thereof as an olfactory receptor OLFR110 agonist. The compound of the present invention is capable of activating OLFR110 in vitro, and binding to OLFR110. The results of animal experiments show that the compound can significantly reduce the body weight of mice and improve blood glucose homeostasis. The compound of the present invention or a pharmaceutically acceptable salt or solvate thereof can be used as a potential pharmaceutical lead compound for treating obesity and diabetes.

Description

A method for preparing a compound and its application as an olfactory receptor OLFR110 agonist. Technical Field

[0001] This invention belongs to the field of medicinal chemistry, specifically relating to a method for preparing a compound and its application as an agonist of the olfactory receptor OLFR110. Background Technology

[0002] The information disclosed in this background section is intended only to enhance understanding of the overall background of the invention and is not necessarily to be construed as an admission or in any way implying that such information constitutes prior art known to those skilled in the art.

[0003] G protein-coupled receptors (GPCRs) are the largest family of cell surface receptor proteins in eukaryotes. Olfactory receptors (ORs) are the largest subfamily of GPCRs. Besides sensing odor molecules, olfactory receptors also exhibit non-olfactory functions, including cell growth, differentiation and apoptosis, migration, and secretion. GPCRs are a very important class of drug targets. To date, the U.S. Food and Drug Administration (FDA) has approved a total of 475 drugs targeting GPCRs, accounting for 33.3% of all FDA-approved drugs. Research on GPCRs has been awarded 11 Nobel Prizes.

[0004] OLFR110 is an orphan GPCR of the class A family, and no known endogenous ligand has been found to date. OLFR110 is highly expressed in olfactory epithelium, liver, and BAT. Studies have found that OLFR110 is a potential new target for the treatment of diabetes. Therefore, the discovery of small molecule agonists of OLFR110 holds promise for a new approach to treating diabetes. Summary of the Invention

[0005] This invention provides a method for preparing a compound and its application as an agonist of the olfactory receptor OLFR110. The compound of this invention can activate the activity of OLFR110 in vitro and can bind to OLFR110. Animal experiments show that the compound can significantly reduce body weight and improve glycemic homeostasis in mice. The compound of this invention, and its pharmaceutically acceptable salts or solvates, can serve as potential drug lead compounds for the treatment of obesity and diabetes.

[0006] In a first aspect, the present invention provides the use of a compound as an agonist of the olfactory receptor OLFR110, the compound being named HOR1-C59, and having the following structural formula:

[0007] In another aspect, the present invention provides the application of HOR1-C59 in activating the Gs signaling path of OLFR110.

[0008] In another aspect, the present invention provides the use of a compound in the preparation of a medicament for the prevention and / or treatment of related glucose and lipid metabolism diseases, the structural formula of which is shown below:

[0009] Furthermore, HOR1-C59 has been used to improve ectopic lipid deposition, enhance energy metabolism, and reduce weight.

[0010] Furthermore, the application of HOR1-C59 in the treatment of obesity.

[0011] Furthermore, HOR1-C59 has been applied to improve glucose tolerance and insulin sensitivity, and enhance the body's ability to regulate blood glucose levels.

[0012] Furthermore, the application of HOR1-C59 in the treatment of diabetes.

[0013] Furthermore, the aforementioned applications include compound HOR1-C59 and its enantiomers, as well as pharmaceutically acceptable salts, solvent compounds, hydrates, or crystals.

[0014] In another aspect, this invention provides a synthetic route for compound HOR1-C59, which is simple, convenient, has a high yield, and uses readily available reactants. The synthetic route for this compound is shown below:

[0015] Furthermore, the specific synthesis method is as follows:

[0016] (-)-Dimethyl-2,3-o-isopropylidene-L-tartrate was dissolved in benzylamine and stirred overnight in an oil bath at 80°C. The reaction solution was diluted with a small amount of water, and the pH was adjusted to 2-3 with 1M HCl solution. The solution was extracted with ethyl acetate, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and the organic phase was concentrated and purified by silica gel column chromatography to give a pale yellow oil, HOR1-C59.

[0017] Compared with the prior art, the beneficial effects of the present invention are:

[0018] The compound of this invention is the first to be found to have OLFR110 agonist activity, is the first small molecule agonist of OLFR110, and is also the first small molecule compound that targets OLFR110 to improve obesity and diabetes.

[0019] This work is of great value in the field of glucose and lipid metabolism. Through structure-based design and screening of small molecule ligands, it provides a potential strategy to improve glucose and lipid metabolism. The relevant results are of great significance to the pharmacological research of olfactory GPCRs and the development of drugs to improve glucose and lipid metabolism. Attached Figure Description

[0020] The accompanying drawings, which form part of this invention, are used to provide a further understanding of the invention. The illustrative embodiments of the invention and their descriptions are used to explain the invention and do not constitute an improper limitation of the invention.

[0021] Figure 1: Effect of compounds on the activity of OLFR110;

[0022] Figure 2: Monitoring of mouse body weight, food intake, and total energy expenditure by compound HOR1-C59;

[0023] Figure 3: Effects of compound HOR1-C59 on glucose tolerance, insulin tolerance and pyruvate tolerance in mice;

[0024] Figure 4: Effects of compound HOR1-C59 on triglyceride levels and adipocyte area in mouse liver;

[0025] Figure 5: H spectrum confirming the structure of compound HOR1-C59. Detailed Implementation

[0026] The embodiments of the present invention are described in detail below. These embodiments are implemented based on the technical solution of the present invention, and provide detailed implementation methods and specific operation processes. However, the scope of protection of the present invention is not limited to the following embodiments.

[0027] Effect of Compound 1 on OLFR110 Activity

[0028] Experimental Methods: The compounds described in this invention were obtained from a vendor in the SPECS compound database (https: / / www.specs.net / ). The activation of the downstream Gs protein of OLFR110 under compound stimulation was then detected using a Gs dissociation assay. HEK293 cells were transiently co-transfected with OLFR110 and the G protein BRET probe. Twenty-four hours after transfection, cells were seeded at 5 × 10⁶ cells per well. 4Cells were aliquoted into 96-well microplates at a density of 1 / 2 cells and incubated at 37°C for 24 hours. Cells were washed twice with Tyrode's buffer (140 mM NaCl, 2.7 mM KCl, 1 mM CaCl2, 12 mM NaHCO3, 5.6 mM D-glucose, 0.5 mM MgCl2, 0.37 mM NaH2PO4, and 25 mM HEPES, pH 7.4) and stimulated with the compounds of this invention at different concentrations. After adding coelentrin (final concentration 5 μM), the BRET signal was measured using a Mithras LB940 microplate reader equipped with a BRET filter set. The BRET signal was calculated as the ratio of emitted light at 510 nm to 400 nm (G protein dissociation assay).

[0029] Experimental results: Further activity tests were conducted on the small molecules obtained through screening, as shown in Figure 1. Through G protein signaling pathway detection and analysis, the EC50 of HOR1-C59 was 7.1±1.6 nM, confirming that the small molecule HOR1-C59 exhibited a strong Gs agonist effect on OLFR110.

[0030] Example 2: Experimental method for monitoring mouse body weight, food intake, and total energy expenditure using compound HOR1-C59: WT and Olfr110... - / - Male mice were intraperitoneally administered either the control solvent or HOR1-C59 (50 mg). -1 Food intake, weight change, and total energy expenditure of 15-week-old male mice were monitored per day (ip) using the CLAMS-16 laboratory animal monitoring system (Columbus Instruments, Columbus, OH, USA).

[0031] Experimental results: As shown in Figure 2, using HOR1-C59 (50ngg) -1 Following per-day (ip) administration, in both the WT group and Olfr110- / - mice, without affecting food intake, the WT mice showed significant weight loss and a significant increase in energy expenditure at night (the gray box in the figure); Olfr110 - / - No improvement in body weight or energy expenditure was observed in mice after administration. This indicates that HOR1-C59 improves systemic metabolism in mice by acting on the Olfr110 receptor.

[0032] Example 3: Effects of compound HOR1-C59 on glucose tolerance, insulin tolerance, and pyruvate tolerance in mice.

[0033] Experimental methods: 8-week-old male mice were subjected to HOR1-C59 (50mg) -1After two months of treatment with either per day (ip) or control solvent, mice were starved for 16 hours before the glucose tolerance test. The plasma glucose levels in the tail vein were measured using a FreeStyle Lite blood glucose meter (Roche) before (baseline) and at 15, 30, 60, 90, and 120 minutes after intraperitoneal injection of 2 mg / g glucose (prepared with physiological saline). For the insulin tolerance test, mice were starved for 6 hours before the insulin tolerance test. The plasma glucose levels in the tail vein were measured using a FreeStyle Lite blood glucose meter (Roche) before (baseline) and at 15, 30, 60, and 120 minutes after intraperitoneal injection of 0.75 mU / g insulin (prepared with physiological saline). For the pyruvate tolerance test, mice were starved for 16 hours before the pyruvate tolerance test. The plasma glucose levels in the tail vein were measured using a FreeStyle Lite blood glucose meter (Roche) before (baseline) and at 15, 30, 60, and 120 minutes after intraperitoneal injection of 2.5 mg / g sodium pyruvate (prepared with physiological saline).

[0034] Experimental results: As shown in Figure 3, after HOR1-C59 (50ngg) -1 After treatment with per day, ip, glucose tolerance (GTT), insulin tolerance (ITT), and pyruvate tolerance (PTT) were significantly improved in WT mice. Olfr110 - / - The glucose tolerance (GTT), insulin tolerance (ITT), and pyruvate tolerance (PTT) of mice were not improved, indicating that compound HOR1-C59 can improve blood glucose homeostasis and increase insulin sensitivity in mice through Olfr110.

[0035] Example 4: Effects of compound HOR1-C59 on triglyceride levels and adipocyte area in mouse liver

[0036] Experimental method: HOR1-C59 (50ngg) -1 After 2 months of treatment with intraperitoneal administration of either per day (ip) or control solvent, WT mice and OLFR110 mice were tested. - / - Frozen sections of mouse liver (10 μM) and paraffin sections of white adipose tissue (10 μM) were stained with ORO (Beyotime) and HE (Solarbio), respectively. Images were captured and processed using a high-resolution panoramic scanning and image analysis system for pathological sections.

[0037] Experimental results: As shown in Figure 4, after treatment with compound HOR1-C59 (50mg / L), -1 After treatment with per day (ip), compared with the control group, WT mice showed a significant reduction in lipid droplets and a significant decrease in the area of ​​white adipocytes in liver slices; Olfr110 - / Compared with the control group, the number of lipid droplets in liver slices was not reduced, and the area of ​​white adipocytes was not decreased. This indicates that HOR1-C59 can effectively improve lipid metabolism in the liver and fat through Olfr110.

[0038] Example 5: Preparation method of compound HOR1-C59

[0039] Experimental method: (-)-dimethyl-2,3-o-isopropylidene-L-tartrate was dissolved in benzylamine, stirred overnight in an oil bath at 80°C, diluted with a small amount of water, and the pH was adjusted to 2-3 with 1M HCl solution. The mixture was extracted with ethyl acetate, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, concentrated the organic phase, and purified by silica gel column chromatography to obtain a pale yellow oil with a yield of 94.1%. 1 H NMR (600MHz, DMSO-d6) δ8.69,8.68,8.67(s,3H),7.33-7.22(m,10H),4.59(s,1H),4.35-4.29(m,4H),1.40(s,6H).

[0040] Obviously, those skilled in the art can make various modifications and variations to this invention without departing from its spirit and scope. Therefore, if these modifications and variations fall within the scope of the claims of this invention and their equivalents, this invention also intends to include these modifications and variations.

Claims

1. The application of a compound as an agonist of the olfactory receptor OLFR110, named HOR1-C59, with the following structural formula:

2. The application as described in claim 1, characterized in that, Application of the compound in activating the Gs signaling pathway of OLFR110.

3. The use of a compound in the preparation of a medicament for the prevention and / or treatment of related glucose and lipid metabolism diseases, the compound having the following structural formula:

4. The application as described in claim 3, characterized in that, Applications of the compound in improving ectopic lipid deposition, enhancing energy metabolism, or reducing weight.

5. The application as described in claim 3, characterized in that, Application of compounds in the treatment of obesity.

6. The application as described in claim 3, characterized in that, Applications of the compound in improving glucose tolerance and insulin sensitivity, and enhancing the body's ability to regulate blood glucose concentration.

7. The application as described in claim 3, characterized in that, Application of compounds in the treatment of diabetes.

8. The application as described in claim 1 or 3, characterized in that, The compounds include compound HOR1-C59 and its enantiomers, as well as pharmaceutically acceptable salts, solvent compounds, hydrates, or crystals.

9. A synthetic route for a compound HOR1-C59, characterized in that, The synthetic route of the compound is shown below:

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