Preparation method of compound and application of compound as olfactory receptor OLFR110 agonist

Abstract

The invention provides a preparation method of a compound and application of the compound as an olfactory receptor OLFR110 agonist. The compound can activate the activity of OLFR110 in vitro and can be combined with the OLFR110, and animal experiment results show that the compound can significantly reduce the weight of mice and improve the blood glucose homeostasis. The compound and the pharmaceutically acceptable salt or solvate of the compound can be used as a potential medicine lead compound for treating obesity and diabetes mellitus.

Description

Technical Field

[0001] The invention belongs to the field of medicinal chemistry, and specifically relates to a preparation method of a compound and an application of the compound as an olfactory receptor OLFR110 agonist. Background Art

[0002] The information disclosed in this background technology section is only intended to enhance the understanding of the overall background of the invention, and should not necessarily be regarded as an admission or any form of suggestion that the information constitutes the prior art already known to a person 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 G protein-coupled receptors (GPCRs). In addition to 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 drugs approved by the FDA. Research on GPCRs has won 11 Nobel Prizes.

[0004] OLFR110 is an orphan GPCR of the class A family, and no known endogenous ligand has been found so far. OLFR110 is highly expressed in the 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 is expected to become a new method for the treatment of diabetes. Summary of the invention

[0005] The present invention provides a method for preparing a compound and its use as an olfactory receptor OLFR110 agonist. The compound of the present invention can activate the activity of OLFR110 in vitro and can bind to OLFR110. Animal experimental results show that the compound can significantly reduce the weight of mice and improve blood sugar homeostasis. The compound of the present invention and its pharmaceutically acceptable salt or solvate can be used as a potential drug lead compound for treating obesity and diabetes.

[0006] In the first aspect of the present invention, a compound is provided for use as an olfactory receptor OLFR110 agonist. The compound is named: HOR1-C59, and the structural formula is as follows:

[0007]

[0008] In another aspect, the present invention provides the use of HOR1-C59 in activating the Gs signaling pathway of OLFR110.

[0009] In another aspect, the present invention provides a compound for use in preparing a drug for preventing and / or treating related glucose and lipid metabolism diseases, the structural formula of which is shown below:

[0010]

[0011] Furthermore, HOR1-C59 is used to improve ectopic lipid deposition, enhance energy metabolism, and reduce body weight.

[0012] Furthermore, HOR1-C59 is used in the treatment of obesity.

[0013] Furthermore, HOR1-C59 is used to improve glucose tolerance and insulin sensitivity, and enhance the body's ability to regulate blood glucose concentration.

[0014] Furthermore, HOR1-C59 is used in the treatment of diabetes.

[0015] Furthermore, the above-mentioned application comprises compound HOR1-C59 and its enantiomers and pharmaceutically acceptable salts, solvates, hydrates or crystals.

[0016] In another aspect, the present invention provides a synthetic route for compound HOR1-C59, which has the characteristics of being simple and convenient, having a high yield, and having readily available reactants. The synthetic route for the compound is as follows:

[0017]

[0018] Further, the specific synthesis method is as follows:

[0019] Dissolve (-)-dimethyl-2,3-o-isopropylidene-L-tartrate with benzylamine, stir in an oil bath at 80°C overnight, dilute the reaction solution with a small amount of water, adjust the pH to 2-3 with 1M HCl solution, extract with ethyl acetate, wash with saturated sodium chloride solution, dry with anhydrous sodium sulfate, concentrate the organic phase and purify with silica gel column chromatography to obtain light yellow oil HOR1-C59.

[0020] Compared with the prior art, the present invention has the following beneficial effects:

[0021] The compound of the present invention is the first to be found to have agonistic activity of OLFR110, is the first small molecule agonist of OLFR110, and is also the first small molecule compound targeting OLFR110 to improve obesity and diabetes.

[0022] This work has important value in the field of lipid metabolism. Through structure-based design and screening of small molecule ligands, it provides a potential strategy to improve lipid metabolism. The related results are of great significance to the pharmacological research of olfactory GPCRs and the development of drugs to improve lipid metabolism. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] The accompanying drawings in the specification, which constitute a part of the present invention, are used to provide a further understanding of the present invention. The exemplary embodiments of the present invention and their descriptions are used to explain the present invention and do not constitute improper limitations on the present invention.

[0024] Figure 1 : Effects of compounds on OLFR110 activity;

[0025] Figure 2 :Compound HOR1-C59 monitors the weight, food intake, and total body energy expenditure of mice;

[0026] Figure 3 :Effects of compound HOR1-C59 on glucose tolerance, insulin tolerance and pyruvate tolerance in mice;

[0027] Figure 4 :Effects of compound HOR1-C59 on triglyceride levels and adipocyte area in mouse liver;

[0028] Figure 5 : H spectrum of compound HOR1-C59 structure confirmation. DETAILED DESCRIPTION

[0029] The following is a detailed description of an embodiment of the present invention. This embodiment is implemented on the premise of the technical solution of the present invention, and a detailed implementation method and a specific operation process are given, but the protection scope of the present invention is not limited to the following embodiment.

[0030] Example 1 Effects of compounds on OLFR110 activity

[0031] Experimental method: The compounds described in the present invention were purchased from suppliers in the SPECS compound database (https: / / www.specs.net / ). The Gs dissociation experiment was then used to detect the activation of the Gs protein downstream of OLFR110 under compound stimulation. HEK293 cells were transiently co-transfected with OLFR110 and G protein BRET probe. After transfection for 24 hours, 5×10 4The cells were distributed into 96-well microplates at a density of 10 cells and incubated at 37°C for 24 hours. The 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 the present invention at different concentrations. After adding coelenterazine (final concentration of 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 the emitted light at 510 nm / 400 nm (G protein dissociation experiment).

[0032] Experimental results: The small molecules obtained by screening were further tested for their activity, such as Figure 1 As shown, through G protein signaling pathway detection analysis, the EC50 of HOR1-C59 was 7.1±1.6nM, confirming that the HOR1-C59 small molecule exhibited a strong Gs agonist effect on OLFR110.

[0033] Example 2 Monitoring of the Effect of Compound HOR1-C59 on Mouse Body Weight, Mouse Food Intake, and Whole Body Energy Expenditure Experimental Method: WT and Olfr110 - / - Male mice were intraperitoneally administered with a control solvent or HOR1-C59 (50 ng g -1 The food intake, body weight changes, and whole-body energy expenditure of 15-week-old male mice were monitored using the Laboratory Animal Comprehensive Monitoring System (CLAMS-16, Columbus Instruments, Columbus, OH, USA).

[0034] Experimental results: Figure 2 As shown, HOR1-C59 (50 ng g -1 After administration of ip per day, the weight of WT mice was significantly reduced, and the energy consumption was significantly increased at night (gray box in the figure) while the food intake of Olfr110- / - mice was not affected. - / - The body weight and energy consumption of mice were not improved after administration, indicating that HOR1-C59 improved the systemic metabolism of mice by acting on the Olfr110 receptor.

[0035] Example 3 Effects of Compound HOR1-C59 on Glucose Tolerance, Insulin Tolerance and Pyruvate Tolerance in Mice Experimental Methods: Male mice aged 8 weeks were treated with HOR1-C59 (50 ng / mL) -1After being treated with 1% paraformaldehyde (2% dapoxetine per day, ip) or control solvent for 2 months, the mice were starved for 16 hours before glucose tolerance test, and the plasma glucose levels in the tail vein blood of the mice were measured with a FreeStyle Lite blood glucose meter (Roche) before (baseline) and 15, 30, 60, 90, and 120 minutes after intraperitoneal injection of 2 mg / g glucose (prepared in normal saline); the mice were starved for 6 hours before insulin tolerance test, and the plasma glucose levels in the tail vein blood of the mice were measured with a FreeStyle Lite blood glucose meter (Roche) before (baseline) and 15, 30, 60, and 120 minutes after intraperitoneal injection of 0.75 mU / g insulin (prepared in normal saline); the mice were starved for 16 hours before pyruvate tolerance test, and the plasma glucose levels in the tail vein blood of the mice were measured with a FreeStyle Lite blood glucose meter (Roche) before (baseline) and 15, 30, 60, and 120 minutes after intraperitoneal injection of 2.5 mg / g sodium pyruvate (prepared in normal saline).

[0036] Experimental results: Figure 3 As shown, HOR1-C59 (50ng -1 per day, ip) treatment, glucose tolerance (GTT), insulin tolerance (ITT), and pyruvate tolerance (PTT) of WT mice were significantly improved, while 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 enhance insulin sensitivity in mice through Olfr110.

[0037] Example 4 Effect of compound HOR1-C59 on triglyceride level and adipocyte area in mouse liver Experimental method: HOR1-C59 (50ng -1 per day, ip) or control solvent were intraperitoneally administered for 2 months to WT mice and OLFR110 - / - Mouse liver frozen sections (10 μM) and white adipose tissue paraffin sections (10 μM) were stained with ORO (Beyotime) and HE (Solarbio), respectively. High-definition pathological section panoramic scanning and image analysis system was used for image capture and processing.

[0038] Experimental results: Figure 4 As shown, the compound HOR1-C59 (50ng -1 per day, ip) treatment, the lipid droplets in liver sections of WT mice were significantly reduced compared with those in the control group, and the area of ​​white adipocytes was significantly reduced; Olfr110 - / -Compared with the control group, the lipid droplets in liver sections did not decrease, and the area of ​​white adipocytes did not decrease, indicating that HOR1-C59 can improve lipid metabolism in liver and fat through Olfr110.

[0039] Example 5 Preparation of Compound HOR1-C59

[0040]

[0041] Experimental method: (-)-dimethyl-2,3-o-isopropylidene-L-tartrate was dissolved with benzylamine, stirred in an oil bath at 80°C overnight, a small amount of water was added to dilute the reaction solution, the pH was adjusted to 2-3 with 1M HCl solution, 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 obtain a light yellow oil with a yield of 94.1%. 1 HNMR (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).

[0042] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include these modifications and variations.

Claims

1. A compound is used as an agonist of olfactory receptor OLFR110. The compound is named: HOR1-C59, and the structural formula is as follows:

2. The use according to claim 1, characterized in that: Application of compounds in activating the Gs signaling pathway of OLFR110.

3. Use of a compound in the preparation of a drug for preventing and / or treating related glucose and lipid metabolism diseases, the compound structural formula is as follows:

4. The use according to claim 3, characterized in that: Use of the compound in improving ectopic lipid deposition, or enhancing energy metabolism, or reducing body weight.

5. The use according to claim 3, characterized in that: Use of compounds in the treatment of obesity.

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

7. The use according to claim 3, characterized in that: Use of the compound in the treatment of diabetes.

8. The use according to claim 1 or 3, characterized in that: The compound includes compound HOR1-C59 and its enantiomers and pharmaceutically acceptable salts, solvates, hydrates or crystals.

9. A synthetic route of compound HOR1-C59, characterized in that: The synthetic route of the compound is as follows:

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