A YDT compound, its preparation method and application

Abstract

This invention relates to the pharmaceutical field, specifically to a YDT compound, its preparation method, and its applications. The invention provides a YDT compound or its pharmaceutically acceptable salt or metabolite, wherein the YDT compound has the structure of Formula 1. Formula 1. Using SAR405 as the core skeleton, this invention aims to enhance its absorption in vivo or reduce its clearance rate by modifying and replacing its ring structure and substituents on the ring, ultimately providing a novel compound with a different skeleton structure and modifying groups. This compound can serve as a host-targeted antiviral drug that blocks viral replication by inhibiting virus-host interactions, and exhibits in vivo antiviral capabilities that SAR405 lacks. Therefore, it has applications in the preparation of VPS34 autophagy inhibitors, anti-SARS-CoV-2 drugs, and drugs for treating pneumonia caused by SARS-CoV-2.

Description

Technical Field

[0001] This invention relates to the field of pharmaceuticals, specifically to a YDT compound, its preparation method, and its applications. Background Technology

[0002] COVID-19, caused by the novel coronavirus, is an acute respiratory infectious disease resulting from SARS-CoV-2, which has caused a major public health crisis worldwide. Currently, treatments for COVID-19 mainly include antiviral drugs, immunomodulators, and corticosteroids, but these still face challenges such as limited efficacy, significant side effects, and the development of drug resistance. Current treatment regimens generally employ anti-SARS-CoV-2 drugs such as nelmatvir / ritonavir combination packs, azvudine tablets, and monorapvir capsules. These three drugs target the virus's own proteins to block viral replication. Currently, there are no host-targeting antiviral drugs used clinically to treat COVID-19.

[0003] Specifically, the main mechanisms of action of anti-SARS-CoV-2 drugs can be divided into two aspects: targeting the virus itself by directly acting on the virus's own proteins or enzymes, interfering with the virus's life cycle, and thus inhibiting viral replication and spread; and targeting the host by regulating key factors or signaling pathways in the host cell, creating an intracellular environment unfavorable to viral replication, thereby indirectly inhibiting viral replication and spread. The former usually has high specificity and can precisely target the key functional proteins of SARS-CoV-2. However, as SARS-CoV-2 continues to evolve, the structure of its own replication enzymes may change accordingly, leading to a decrease in the efficacy of traditional antiviral drugs that target viral proteins. The latter directly targets the key proteins required for viral replication, exhibiting a broader spectrum of anti-SARS-CoV-2 capabilities.

[0004] VPS34 (Vacuolar Protein Sorting 34), a key protein involved in SARS-CoV-2 replication, holds promise as a novel drug target for anti-SARS-CoV-2 treatments. VPS34 is a class III phosphatidylinositol 3-kinase (PI3K-III) that plays a central role in cellular processes such as autophagy, endocytosis, and vesicle transport. Studies have shown that after SARS-CoV-2 infects host cells, it relies on VPS34's autophagy-related functions to form double-membrane vesicles (DMVs) to create replication sites. Therefore, inhibiting VPS34 may be a potential strategy against SARS-CoV-2, making host-targeted VPS34 autophagy inhibitors a hot research topic. Regarding the use of autophagy inhibitors in anti-coronavirus treatment, autophagy inhibitors such as chloroquine and hydroxychloroquine have been attempted for COVID-19 treatment. They indirectly inhibit autophagy by increasing lysosomal pH, but clinical studies have shown limited efficacy and significant side effects.

[0005] Currently, only a few studies have reported that VPS34 inhibitors VPS34 IN-1, VPS34 IN-2, SAR405, and Autophinib can inhibit SARS-CoV-2 replication in in vitro models. No studies have shown that these inhibitors have significant inhibitory activity against SARS-CoV-2 in vivo. In particular, SAR405, despite being the best existing VPS34 inhibitor, has been found to have poor anti-SARS-CoV-2 efficacy in mice. Summary of the Invention

[0006] In view of this, the technical problem to be solved by the present invention is a YDT compound, its preparation method and application. The YDT compound provided by the present invention has excellent ability to inhibit VPS34 autophagy against SARS-CoV-2 in vivo, and can be used as a host-targeted antiviral drug for the clinical treatment of COVID-19.

[0007] This invention provides a YDT compound or a pharmaceutically acceptable salt or metabolite thereof, wherein the YDT compound has the structure of Formula 1;

[0008] Formula 1.

[0009] The YDT compound provided by this invention is a brown powdery solid with a solubility of approximately 0.12 mg / mL in water and greater than 50 mg / mL in DMSO, exhibiting low hygroscopicity. Using SAR405 as the core framework, the inventors modified and replaced its ring structure and substituents to enhance its in vivo absorption and reduce its clearance rate, ultimately providing a novel compound named YDT. This compound can serve as a host-targeted antiviral drug that blocks viral replication by inhibiting virus-host interactions, demonstrating good antiviral activity in vivo.

[0010] This invention also provides a method for preparing a YDT compound, comprising the following steps:

[0011] The compound having the structure of formula A was reacted with 3-methylmorpholine and 1-amino-3,3-dimethylbut-2-one to give the YDT compound having the structure of formula 1.

[0012] Formula A; Formula 1.

[0013] This invention uses a compound having the structure of formula A as the main skeleton, and reacts it with 3-methylmorpholine and 1-amino-3,3-dimethylbut-2-one, wherein the structure of 3-methylmorpholine is as follows: The structure of the 1-amino-3,3-dimethylbut-2-one is as follows: By replacing the Cl group on the skeleton of SAR405 with 3-methylmorpholine, a skeleton structure similar to but different from SAR405 was constructed. By introducing a new modifying group at the amino site that originally belonged to SAR405 with 1-amino-3,3-dimethylbut-2-one, the chemical structure of that site was adjusted. By combining the optimization of the skeleton structure and the modifying group, the YDT compound described in this invention was obtained.

[0014] This invention can first synthesize the compound having the structure of formula A, and then use it to further prepare the YDT compound having the structure of formula 1, specifically including the following steps:

[0015] S1) Reacting a compound having the structure of formula A-0 with 3-oxopropionic acid yields a compound having the structure of formula A-1;

[0016] Formula A-0; Formula A-1;

[0017] S2) React the compound with the structure of formula A-1 obtained in step S1) with phosphorus oxychloride to obtain a compound with the structure of formula A;

[0018] S3) The compound with the structure of formula A obtained in step S2) is reacted with 3-methylmorpholine and 1-amino-3,3-dimethylbut-2-one to obtain the YDT compound with the structure of formula 1.

[0019] In some embodiments of the present invention, the preparation method of the YDT compound specifically includes the following steps: S1) under the action of DCC and HoBt, reacting a compound having the structure of formula A-0 and 3-oxopropionic acid in an organic solvent to obtain a compound having the structure of formula A-1; S2) under the action of triethylamine, reacting the compound having the structure of formula A-1 obtained in step S1) with phosphorus oxychloride to obtain a compound having the structure of formula A; S3) reacting the compound having the structure of formula A obtained in step S2), 3-methylmorpholine and 1-amino-3,3-dimethylbut-2-one in ethanol to obtain a YDT compound having the structure of formula 1.

[0020] In the preparation method of the YDT compound of the present invention, the reaction temperature in step S1) is room temperature, specifically 15℃~30℃; the reaction time in step S1) is 10 h~15 h, preferably 12 h. The reaction temperature in step S2) is 80℃~100℃, preferably 90℃; the reaction time in step S2) is 10 h~15 h, preferably 12 h. The reaction temperature in step S3) is room temperature, specifically 15℃~30℃, preferably 20℃~25℃; the reaction time in step S3) is 4 h~8 h, preferably 6 h.

[0021] This invention provides the use of any of the YDT compounds described above, or their pharmaceutically acceptable salts, metabolites, or YDT compounds obtained by any of the preparation methods described above, in the preparation of drugs that inhibit VPS34 autophagy. The YDT compounds provided by this invention are optimized based on SAR405, and like SAR405, they have the effect of inhibiting VPS34 autophagy. The difference is that the YDT compounds provided by this invention also have the effect of inhibiting VPS34 autophagy in vivo. Preparing the YDT compounds described in this invention into VPS34 autophagy inhibitors can resist viruses that replicate based on VPS34 autophagy.

[0022] This invention also provides the application of any of the YDT compounds described above, or their pharmaceutically acceptable salts, metabolites, or YDT compounds obtained by any of the preparation methods described above, in the preparation of drugs against SARS-CoV-2 virus. Specifically, after SARS-CoV-2 infects host cells, it relies on VPS34 autophagy-related functions to form double-membrane vesicle structures to create replication sites. The YDT compounds of this invention can inhibit VPS34 autophagy, thereby exhibiting excellent anti-SARS-CoV-2 virus activity, and are therefore very suitable for use in the preparation of drugs against SARS-CoV-2 virus.

[0023] This invention also provides the use of any of the YDT compounds described above, or their pharmaceutically acceptable salts, metabolites, or YDT compounds obtained by any of the preparation methods described above, in the preparation of medicaments for treating pneumonia caused by SARS-CoV-2 virus. Further investigation of the pharmacokinetics of SAR405 revealed that SAR405 exhibits poor absorption in vivo and poor distribution in the lungs, the target organ of SARS-CoV-2, which may be the main reason for its lack of antiviral effect in vivo. The YDT compounds obtained by optimizing the structure of SAR405 enhance in vivo absorption and reduce in vivo clearance rates, resulting in superior therapeutic effects against pneumonia caused by SARS-CoV-2 virus, making them highly suitable for use in the preparation of medicaments for treating pneumonia caused by SARS-CoV-2 virus.

[0024] This invention also provides a pharmaceutical formulation comprising an active compound and excipients; the active compound is selected from any of the YDT compounds described above or their pharmaceutically acceptable salts, metabolites, or YDT compounds obtained by any of the preparation methods described above. This invention does not specifically limit the excipients, and any excipient acceptable for pharmaceutical use is acceptable. The dosage form of the pharmaceutical formulation of this invention is an oral formulation or an injectable formulation.

[0025] This invention provides a YDT compound, its preparation method, and its application. Specifically, this invention provides a YDT compound or a pharmaceutically acceptable salt or metabolite thereof, wherein the YDT compound has the structure of Formula 1; Formula 1. This invention uses SAR405 as the core skeleton, aiming to enhance its absorption in vivo or reduce its clearance rate. It modifies and replaces its ring structure and substituents on the ring, ultimately providing a novel compound with a different skeleton structure and modified groups, named YDT compound. This compound can serve as a host-targeted antiviral drug that blocks viral replication by inhibiting virus-host interactions, exhibiting in vivo antiviral capabilities that SAR405 lacks. Therefore, it has applications in the preparation of VPS34 autophagy inhibitors and in the preparation of drugs against SARS-CoV-2 virus, and can treat pneumonia caused by SARS-CoV-2 virus, thus having applications in the preparation of drugs for treating pneumonia caused by SARS-CoV-2 virus. Attached Figure Description

[0026] Figure 1 The graph shows the cellular half-maximal toxicity concentration (IC50) of SAR405.

[0027] Figure 2 The graph shows the cytotoxicity concentration (IC50) of YDT.

[0028] Figure 3 A graph comparing the in vivo antiviral activity and survival rate of SAR405 and YDT;

[0029] Figure 4 A graph comparing the antiviral capabilities and viral load of SAR405 and YDT in vivo;

[0030] Figure 5 A comparison of the N gene copy number in vivo to assess the antiviral capabilities of SAR405 and YDT. Detailed Implementation

[0031] This invention discloses a YDT compound, its preparation method, and its applications. Those skilled in the art can refer to this document and appropriately modify the process parameters to achieve the desired results. It should be particularly noted that all similar substitutions and modifications are obvious to those skilled in the art and are considered to be included in this invention. The methods and applications of this invention have been described through preferred embodiments. Those skilled in the art can clearly modify or appropriately change and combine the methods and applications described herein without departing from the content, spirit, and scope of this invention to realize and apply the technology of this invention.

[0032] The compounds used in this invention 3-O-propionic acid, DMF (N,N-dimethylformamide), DCC (dicyclohexylcarbodiimide), HoBt (1-hydroxybenzotriazole), phosphorus oxychloride, triethylamine, ethanol, and 1-amino-3,3-dimethylbut-2-one are all commercially available reagents.

[0033] The present invention will be further described below with reference to the embodiments:

[0034] Example 1

[0035] The YDT compound was synthesized according to the following reaction formula:

[0036]

[0037] The specific process is as follows:

[0038] Step 1: Put Dissolve 100 mM (18.56 g) and 3-oxopropionic acid (120 mM, 10.56 g) in 800 mL of DMF, add DCC (120 mM, 24.72 g) and HoBt (120 mM, 16.2 g), and stir at room temperature (20–25 °C) for 12 hours to obtain 20.48 g of pale yellow solid intermediate B; yield 80.95%; purity 94%.

[0039] Step 2: Intermediate B (20 mmol, 5.08 g) was dissolved in phosphorus oxychloride (200 mmol, 30.37 g), and triethylamine (24 mmol, 2.43 g) was added dropwise to the mixture. The mixture was stirred at 90 °C for 12 h. The reaction solution was evaporated to dryness and recrystallized from dichloromethane to obtain 3.78 g of yellow intermediate C; yield 51.20%, purity 95%.

[0040] Step 3: Mix intermediate C (5 mmol, 1.36 g) and (5 mmol, 0.51 g) was dissolved in 100 mL of anhydrous ethanol, stirred at room temperature (20–25 °C) for 6 hours, and then 1-amino-3,3-dimethylbut-2-one (6 mmol, 0.69 g) was added. The mixture was stirred for another 6 hours. After the reaction was completed, the solvent was removed under reduced pressure, and 0.8 g of the white product was obtained by column chromatography. That is, the YDT compound described in this invention has a yield of 41.67% and a purity of 98%.

[0041] The cytotoxicity and in vivo antiviral activity of the YDT compound prepared above were compared with SAR405, as detailed below:

[0042] 1. Comparison of YDT compound and SAR405 cytotoxicity.

[0043] Vero E6 cells were seeded into 96-well plates, with 1 × 10⁶ cells per well. 4After overnight culture, the original culture medium in the 96-well plates was discarded. Different concentrations of YDT and SAR405 were prepared using serum- and antibiotic-free DMDM, and 100 μL of each solution was transferred to each well. Then, 100 μL of DMEM containing 10% FBS was added to each well. The plates were incubated at 37°C with 5% CO2 for 48 hours. Afterward, the culture medium was discarded, and diluted CCK8 assay reagent was added to each well. The plates were incubated at 37°C for 2 hours, and cell viability was calculated using a microplate reader. Data were processed using Graphpad Prism software to determine the corresponding half-maximal cytotoxicity concentration (MCC). 50 ), the result is as follows Figure 1 and Figure 2 As shown, Figure 1 This is a graph showing the cellular half-maximal toxicity (IC50) of SAR405. Figure 2 The graph shows the cytotoxicity concentration (IC50) of YDT. It can be seen that in Vero E6 cells, the drug toxicity of YDT is approximately four times lower than that of SAR405.

[0044] 2. Comparison of the antiviral activity of compound YDT with that of SAR405 mice in vivo.

[0045] (1) Comparison of survival rates

[0046] Eight-month-old Balb / c mice were randomly divided into four groups (n=6 / group): (1) Mock group: PBS was administered intranasally; (2) SARS-CoV-2+Vechicle group (solvent control group): SARS-CoV-2 was administered intranasally, and blank solvent was administered intraperitoneally daily; (3) SARS-CoV-2+SAR405 group: SARS-CoV-2 was administered intranasally, and SAR405 was administered intraperitoneally at 100 mg / kg daily; (4) SARS-CoV-2+YDT group: SARS-CoV-2 was administered intranasally, and YDT was administered intraperitoneally at 100 mg / kg daily. All groups were observed for seven consecutive days after viral infection, and survival curves were calculated. The results are as follows: Figure 3 As shown, Figure 3 The graph shows a comparison of the in vivo antiviral activity and survival rate of SAR405 and YDT. It can be seen that on day 5 post-infection, all mice in the solvent control group and the SAR405 group died, while the survival rate of mice in the YDT-treated group was 33%.

[0047] (2) Comparison of viral load and N gene copy number

[0048] The model and survival curve treatments were consistent (n=3 / group). Mice were sacrificed on day 3 of viral infection, and tissue samples were collected for testing to detect viral replication in lung tissue. The results are as follows: Figure 4 and Figure 5 As shown, Figure 4This is a comparison of the antiviral capabilities and viral load of SAR405 and YDT in vivo. Figure 5 This is a comparison of the N gene copy number in the in vivo antiviral capabilities of SAR405 and YDT mice. It is evident that the viral load in the YDT-treated group was significantly lower than that in the control group, and the corresponding N gene copy number was also reduced.

[0049] The above description is only a preferred embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any equivalent substitutions or modifications made by those skilled in the art within the scope of the technology disclosed in the present invention, based on the technical solution and inventive concept of the present invention, should be covered within the scope of protection of the present invention.

Claims

1. A YDT compound or a pharmaceutically acceptable salt thereof, characterized in that, The YDT compound has a structure of Formula 1. Formula 1.

2. A method of preparing a YDT compound, characterized by, The method comprises the following steps: reacting a compound having a structure of Formula A, 3-methylmorpholine and 1-amino-3,3-dimethylbutan-2-one to obtain a YDT compound having a structure of Formula 1; Formula A; Formula 1.

3. The preparation method according to claim 2, characterized in that, The method comprises the following steps: S1) reacting a compound having a structure of Formula A-0 and 3-oxopropionic acid to obtain a compound having a structure of Formula A-1; Formula A-0; Formula A-1; S2) reacting the compound having a structure of Formula A-1 obtained in step S1) and phosphorus oxychloride under the action of triethylamine to obtain a compound having a structure of Formula A; S3) reacting the compound having a structure of Formula A obtained in step S2), 3-methylmorpholine and 1-amino-3,3-dimethylbutan-2-one in ethanol to obtain a YDT compound having a structure of Formula 1.

4. The production method according to claim 3, characterized by, The method comprises the following steps: S1) reacting a compound having a structure of Formula A-0 and 3-oxopropionic acid in an organic solvent under the action of DCC and HOBt to obtain a compound having a structure of Formula A-1; S2) reacting the compound having a structure of Formula A-1 obtained in step S1) and phosphorus oxychloride under the action of triethylamine to obtain a compound having a structure of Formula A; S3) reacting the compound having a structure of Formula A obtained in step S2), 3-methylmorpholine and 1-amino-3,3-dimethylbutan-2-one in ethanol to obtain a YDT compound having a structure of Formula 1.

5. The production method according to claim 3 or 4, characterized by, The temperature of the reaction in step S1) is 15-30°C, and the time of the reaction in step S1) is 10-15 h; The temperature of the reaction in step S2) is 80-100°C, and the time of the reaction in step S2) is 10-15 h; The temperature of the reaction in step S3) is 15-30°C, and the time of the reaction in step S3) is 4-8 h.

6. Use of the YDT compound of claim 1 or a pharmaceutically acceptable salt thereof or the YDT compound obtained by the preparation method of any one of claims 2-5 in the preparation of a VPS34 autophagy inhibitor.

7. Use of the YDT compound of claim 1 or a pharmaceutically acceptable salt thereof or the YDT compound obtained by the preparation method of any one of claims 2-5 in the preparation of an anti-SARS-CoV-2 virus drug.

8. Use of the YDT compound of claim 1 or a pharmaceutically acceptable salt thereof or the YDT compound obtained by the preparation method of any one of claims 2-5 in the preparation of a drug for treating pneumonia caused by the SARS-CoV-2 virus.

9. A pharmaceutical preparation, characterized in that, The active compound is selected from the YDT compound of claim 1 or a pharmaceutically acceptable salt thereof or the YDT compound obtained by the preparation method of any one of claims 2-5. The dosage form is an oral preparation or an injection preparation.

10. The pharmaceutical preparation according to claim 9, characterized in that, ​

Images