Application of cationic rosin and its citrullus ring complex in the preparation of drugs for preventing or treating novel coronavirus infection
By preparing cationic peach rosé and its melon ring complex, the problem of lack of Cathepsin L inhibitors in the prior art was solved, effective treatment of novel coronavirus infection was achieved, and various drug forms such as tablets, capsules and oral liquids were provided.
Patent Information
- Application Number
- CN202310381493.7
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Filing Date
- 2023-04-11
- Publication Date
- 2025-08-26
- Estimated Expiration
- 2043-04-11
AI Technical Summary
There is no application of cationic peach rosé and its melon ring complex in the prior art for the treatment of novel coronavirus infection, and there is a lack of effective drug targets such as Cathepsin L inhibitors.
Cationic peach roses and melon ring complexes are used to form a complex by reacting the cationic peach roses with the misaligned ten-melon rings with cationic peach roses, which are used to prepare drugs to prevent or treat anti-novel coronavirus infection. Specific dosage forms include tablets, capsules, granules and oral liquids.
The cationic rosé melon ring complex has a significant inhibitory effect on Cathepsin L protease activity, with an inhibition rate of 77.75% to 84.68%, and an IC50 of 438.8μM to 734.7μM, providing an effective anti-free coronavirus infection drug solution.
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Figure CN116763779B_ABST
Abstract
Description
Technical Field
[0001] The present invention relates to the field of compounds, and in particular to the use of cationic rosin and its citrullus ring complex in the preparation of drugs for preventing or treating novel coronavirus infection. Background Art
[0002] The replication cycle of the novel coronavirus (SARS-CoV-2) includes stages such as invasion, replication, assembly, and secretion. Blocking viral replication can occur at any stage of the cycle. The viral genome encodes 28 proteins, including 16 nonstructural proteins, 4 structural proteins, and 8 accessory proteins. Currently established drug targets for the novel coronavirus host include angiotensin-converting enzyme 2 (ACE2), transmembrane serine protease 2 (TMPRSS2), furin, cathepsin L, phosphatidyl inositol 3-phosphate 5-kinase (PIKfyve), protein-associated kinase 1 (AAK1), and cyclin G-associated kinase (GAK). It has been reported that the hydrolysis process of SARS-CoV S protein plays a key role in viral invasion and fusion. Cathepsin L has been shown to be a related exogenous gene protease at the key cleavage site in the enzymatic hydrolysis process of SARS-CoV S protease, which can promote the entry of the virus into host cells. At the same time, SARS-CoV-2 infection also promotes the expression and enzymatic activity of Cathepsin L, thereby enhancing viral infection. Therefore, Cathepsin L is an important therapeutic target for COVID-19. The development of Cathepsin L inhibitors can effectively inhibit viral replication and viral function, thereby interfering with the viral assembly process to make it non-infectious and ultimately prevent viral infection.
[0003] Cationic pink is a dark red, uniform powder that dissolves in water to a vibrant pink. It is a dye used for dyeing and printing acrylic and its blended fabrics. Currently, there is no literature documenting its medicinal use, nor is there any evidence that cationic pink and its cucurbitacin complex can be used to treat novel coronavirus infection. This study investigates the effects of cationic pink and its cucurbitacin complex, confirming their efficacy in treating novel coronavirus infection and expanding the application of cationic pink. Summary of the Invention
[0004] The purpose of this patent is to provide an application of cationic rosin and its cucurbitacin complex in the preparation of drugs for preventing or treating anti-new coronavirus infection, which can be used to prepare drugs for preventing or treating anti-new coronavirus infection.
[0005] The technical solution of the present invention:
[0006] Application of cationic rosin and its citrullus ring complex in the preparation of drugs for preventing or treating novel coronavirus infection.
[0007] The use of the aforementioned cationic pink and its cucurbitacin complex in the preparation of drugs for treating anti-new coronavirus infection, wherein the cationic pink has the molecular formula C 22 H 26 Cl2N2; the structural formula of the cationic pink is:
[0008]
[0009] The use of the aforementioned cationic peach red and its cucurbitacin complex in the preparation of drugs for treating anti-new coronavirus infection, wherein the cationic peach red cucurbitacin complex is obtained by reacting a dislocated ten-membered cucurbitacin ring with cationic peach red; the molecular formula of the cationic peach red complex is: C 80 H 88 N 40 O 20 Cl2N2; the structural formula of the cationic pink complex is:
[0010]
[0011] The preparation method of the aforementioned cationic peach red cucurbit ring complex is: weigh 1.5 mg of dislocated ten-membered cucurbit ring and 0.5 mg of cationic peach red, and mix them to obtain the cationic peach red cucurbit ring complex; the molar ratio of the dislocated ten-membered cucurbit ring to cationic peach red is 1:1.
[0012] Specifically, the preparation method of the aforementioned cationic peach red cucurbit ring complex is: weigh 1.5 mg of the dislocated ten-membered cucurbit ring and 0.5 mg of cationic peach red, add 0.4-2 mL of deionized water, mix and react, and obtain a cationic cucurbit ring peach red complex with a concentration of 1-5 mg / mL; the molar ratio of the dislocated ten-membered cucurbit ring to cationic peach red is 1:1.
[0013] More specifically, the preparation method of the aforementioned cationic peach red cucurbitacin complex is as follows: weigh 1.5 mg of dislocated ten-membered cucurbitacin and 0.5 mg of cationic peach red, add 2 mL of deionized water, mix and react, and obtain a cationic cucurbitacin pink complex with a concentration of 1 mg / mL; the molar ratio of the dislocated ten-membered cucurbitacin to cationic peach red is 1:1.
[0014] The use of the aforementioned cationic rhodamine and its cucurbitacin complex in the preparation of drugs for preventing or treating novel coronavirus infection, wherein the cationic rhodamine and its cucurbitacin complex are combined with acceptable excipients in the drug, processed according to conventional methods, and prepared into the corresponding drug.
[0015] The aforementioned medicines are tablets, capsules, granules, and oral solutions.
[0016] Beneficial effects of the present invention
[0017] When the aqueous solution concentration of cationic rosin and cationic rosin cucurbitacin complex (cationic rosin-ns-Q
[10] ) was 1 mg / ml, the inhibition rates on Cathepsin L protease activity were 77.75% and 84.68%, respectively, with IC50 values of 438.8 μM and 734.7 μM, respectively. They can be used to prepare drugs for anti-novel coronavirus infection.
[0018] The present invention is simple to operate, the cationic cucurbitacin complex (i.e., the cationic cucurbitacin-ns-Q
[10] ) is easy to prepare, has clear medicinal efficacy, and is easy to produce and apply on a large scale. BRIEF DESCRIPTION OF THE DRAWINGS
[0019] Figure 1 :The structural diagram of the dislocated ten-membered cucurbit ring (ns-Q
[10] );
[0020] Figure 2 : is the molecular structure diagram of cationic pink;
[0021] Figure 3 : is the NMR hydrogen spectrum of cationic pink molecule;
[0022] Figure 4 : This is the H NMR spectrum of the interaction between the dislocated ten-membered cucurbit ring and the cation;
[0023] Figure 5 : The inhibition rate of different concentrations of cationic rosin on the main protease of SARS-CoV-2;
[0024] Figure 6 :The inhibition rate of SARS-CoV-2 main protease by different concentrations of cationic pink-ns-Q
[10] ;
[0025] Figure 7 :Protease IC 50 Figure (a) shows the IC of cationic pink against the main protease of SARS-CoV-2 50 Figure; (b) shows the IC of cationic pink-ns-Q
[10] to the main protease of SARS-CoV-2 50 picture). DETAILED DESCRIPTION
[0026] The present invention will be further described below with reference to the examples, but they are not intended to limit the present invention.
[0027] Example 1:
[0028] Preparation method of cationic peach red cucurbit ring complex: weigh 1.5 mg of dislocated ten-membered cucurbit ring and 0.5 mg of cationic peach red, add 2 mL of deionized water, mix and react, and obtain a cationic peach red cucurbit ring complex with a concentration of 1 mg / mL.
[0029] Take the cationic peach red cucurbitacin complex, add appropriate amounts of conventional excipients, mix well, form granules, dry, and compress into tablets. Each tablet contains 1 mg of the cationic peach red cucurbitacin complex.
[0030] Example 2:
[0031] Preparation method of cationic peach red cucurbit ring complex: weigh 1.5 mg of dislocated ten-membered cucurbit ring and 0.5 mg of cationic peach red, add 1 mL of deionized water, mix and react to obtain a cationic peach red cucurbit ring complex with a concentration of 1 mg / mL.
[0032] Take the cationic peach red cucurbitacin complex, add appropriate amount of conventional excipients, mix well, dry, sterilize, put into hard capsules, and package to prepare capsules. Each capsule contains 2 mg of cationic peach red.
[0033] Dosage and Administration: This preparation is taken once a day, 2 tablets each time, for the treatment of novel coronavirus infection.
[0034] Example 3:
[0035] Preparation method of cationic peach red cucurbit ring complex: weigh 1.5 mg of dislocated ten-membered cucurbit ring and 0.5 mg of cationic peach red, add 0.4 mL of deionized water, mix and react to obtain a cationic peach red cucurbit ring complex with a concentration of 5 mg / mL.
[0036] Take the cationic peach red cucurbitacin complex, add an appropriate amount of solubilizer, grind, dilute with a small amount of water, mix thoroughly, add flavoring and preservatives, mix thoroughly, add water to the prescribed amount, filter, mix thoroughly, package, and sterilize to prepare an oral solution. Each bottle of oral solution contains 5 mg of the cationic peach red cucurbitacin complex.
[0037] Dosage and Administration: Take this preparation once a day, 2 bottles each time, for the treatment of novel coronavirus infection.
[0038] Example 3: Cationic pink (purchased from the market) was added with appropriate amounts of conventional excipients, mixed, dried, sterilized, filled into hard capsules, and packaged to prepare capsules. Each capsule contained 1 mg of cationic pink.
[0039] Dosage and Administration: This preparation is taken once a day, 2 capsules each time, for the treatment of novel coronavirus infection.
[0040] Example 4: Cationic rosin (purchased from the market) was added with appropriate amounts of common auxiliary materials to form granules, which were then dried and granulated to prepare granules. The granules were 5 g / bag and contained 5 mg of cationic rosin.
[0041] Dosage and Administration: Take this preparation once a day, 2 bags each time, for the treatment of novel coronavirus infection.
[0042] Example 5:
[0043] Take cationic peach red, add an appropriate amount of solubilizer, grind, dilute with a small amount of water, mix thoroughly, add flavoring agent and preservative, mix thoroughly, add water to the prescribed amount, filter, mix thoroughly, package, and sterilize to prepare an oral solution. The content of cationic peach red cucurbitacin complex in each bottle of oral solution is 2 mg.
[0044] Dosage and Administration: Take this preparation once a day, 2 bottles each time, for the treatment of novel coronavirus infection.
[0045] The present invention has carried out a lot of experimental research, and the following is the result of the experimental research of the present invention:
[0046] 1. Experimental study on the properties of cationic pink and its cucurbitacin complex
[0047] 1.1 Instruments and reagents
[0048] Testing instrument: JEM-1400 Flash nuclear magnetic resonance instrument
[0049] Test drugs: cationic pyrofluorexamine (APFG), cationic pyrofluorexamine cucurbitacin complex (APFG-ns-Q
[10] ), and deuterated water (Aladdin).
[0050] 1.2 Experimental methods and results
[0051] 1.2.1 Test solution preparation:
[0052] Weigh 10 mg of cationic pink and add deuterated water to make 1x10 -2 mol / L solution for use, weigh 10 mg of ns-Q
[10] , add deuterated water to make 1x10 -3 mol / L solution for later use.
[0053] Pipette 25 μl of the prepared cationic pink solution and add it to the NMR tube, then add 475 μl of deuterated water to prepare test tube No. 1.
[0054] Pipette the same volume of cationic pink solution and add it to another NMR tube according to the above method to make test tube No. 2.
[0055] 1.2.2 Test methods and results:
[0056] Place test tube No. 1 directly into the nuclear magnetic resonance instrument and measure its hydrogen spectrum to obtain the following Figure 3 The positive ion red hydrogen spectrum is shown.
[0057] Put the No. 2 test tube directly into the nuclear magnetic resonance instrument and measure its hydrogen spectrum to obtain the following Figure 3 The cationic red hydrogen spectrum is shown. Then 0.2 equivalents of ns-Q
[10] deuterated aqueous solution are added dropwise each time to obtain Figure 4The hydrogen spectrum of the cationic pink complex is shown.
[0058] 1.2.3 Preparation method of cationic peach red cucurbit ring complex:
[0059] 1.5 mg of dislocated ten-membered cucurbitacin and 0.5 mg of cationic peach red were weighed, 2 mL of deionized water was added, and after mixing and reaction, a cationic peach red cucurbitacin complex with a concentration of 1 mg / mL was prepared.
[0060] 2. Drug efficacy evaluation test
[0061] 2.1 Instruments and reagents
[0062] Detection instruments: TECAN multifunctional microplate reader (infinite M200 Pro), 1 / 100,000 electronic analytical balance model: MS205DU (METTLER Totoledo, Switzerland), Aiko ultrapure water machine (AKHL-III-08); Eppendorf pipette.
[0063] Reagent test kit: 520 Cathepsin L Kit *Fluorimetric* (Cat#:AS-72218, Lot#:1014, AnaSpec, San Jose, USA).
[0064] Test drugs: cationic fluorene (APFG), cationic fluorene-ns-Q
[10] (APFG-ns-Q
[10] ), methanol, acetonitrile and formic acid (chromatographic grade, Fisher, USA), and ultrapure water.
[0065] Positive control: Cathpsin L inhibitor (Cat#: AS-72218, Lot#: 171-112, AnaSpec, San Jose, USA).
[0066] 2.2 Cathepsin L protease experimental methods and results:
[0067] 2.2.1 Preparation of test solution:
[0068] Cationic Peach Red-Cucurbitacin Complex Solution: Take 2 mg of cationic Peach Red-Cucurbitacin complex and add 200 μL of deionized water to prepare a 10 mg / mL cationic Peach Red-Cucurbitacin complex aqueous solution. Sequentially dilute the 10 mg / mL cationic Peach Red-Cucurbitacin complex aqueous solution to prepare 5 mg / mL, 1 mg / mL, 0.2 mg / mL, 0.04 mg / mL, and 0.008 mg / mL aqueous solutions.
[0069] Cationic Peach Red Solution: Weigh 2 mg of cationic Peach Red and add 200 μL of deionized water to prepare a 10 mg / mL cationic Peach Red solution. Dilute the 10 mg / mL cationic Peach Red solution to prepare aqueous solutions of 5 mg / mL, 1 mg / mL, 0.2 mg / mL, 0.04 mg / mL, and 0.008 mg / mL, respectively.
[0070] Buffer solution: Mix 2X assay buffer (Component D) and 1M DDT (Component F) at a ratio of 124:1 and keep on ice until ready to use.
[0071] Dissolve Cathepsin L protease substrate: Dilute the substrate (Component A) 100-fold with 2X assay buffer and keep on ice until ready to use.
[0072] Cathepsin L protease solution: Dilute the enzyme (Component C) 200-fold with 2X assay buffer. Prepare fresh and keep on ice until ready for use.
[0073] Cathepsin L inhibitor: Dilute 100 μM Cathepsin L inhibitor (Component E) 100-fold with 2X assay buffer.
[0074] 2.2.2 Determination method:
[0075] To a 384-well black plate (Lot# 26821021, Corning, USA), substrate blank solution (SC), vehicle blank solution (VC), positive control (PC), inhibitor control (IC), and test compound control (TCC) were first added. Cathepsin L protease was then added, followed by substrate (5 mg / mL, 1 mg / mL, 0.2 mg / mL, 0.04 mg / mL, and 0.008 mg / mL of cationic rosin). Separately, substrate (5 mg / mL, 1 mg / mL, 0.2 mg / mL, 0.04 mg / mL, and 0.008 mg / mL of cationic rosin cucurbitacin complex) was prepared and added according to the grouping method described above. Each concentration was plated in triplicate wells, and after oscillation mixing, the cells were incubated at 37°C for 30 minutes. The fluorescence intensity was measured using a TECAN multifunctional microplate reader at an excitation wavelength of 485 nm and an emission wavelength of 525 nm.
[0076] Data processing and analysis: The data were expressed as mean ± standard deviation (X±S), and statistical processing was completed using Microsoft Windows 2021 Excel software. The inhibition rate and half-maximal inhibitory concentration (IC50) were calculated using the STDEV-t test for the quantitative data.
[0077] Inhibition rate calculation formula: Inhibition rate % = 100 × (F vehicle -F sample ) / (F vehicle -F Substrate )
[0078] Where: F vehicle is the fluorescence value of the solvent blank solution;
[0079] F sample is the fluorescence value of the test compound solution;
[0080] F Substrate is the fluorescence value of the substrate blank solution.
[0081] IC50 is the concentration at which B / B0 = 50%, which can be calculated using GraphPad Prism 9.3.0 software by nonlinear fitting. Where: B is the fluorescence intensity of the wells containing the inhibitory substance, and B0 is the fluorescence intensity of the control wells without the inhibitory substance.
[0082] 2.2.3 Results of Cationic Pink Anti-Cathepsin L Protease Activity Screening
[0083] Inhibition rate of cationic rosin on Cathepsin L protease (n=3)
[0084] Table 1 Inhibition rate of cathepsin L by different concentrations of cationic pink
[0085]
[0086]
[0087] Table 2 Inhibition rate of cathepsin L by cathepsin-cucurbitacin complex at different concentrations
[0088]
[0089] The test results are shown in Figure 5 And Table 1: When the aqueous solution concentration of cationic rosin and cationic rosin-ns-Q
[10] is 1 mg / ml, the inhibition rates on Cathepsin L are 77.75% and 84.68%, respectively, and the half inhibition rates IC50 are 438.8 μM and 734.7 μM, respectively, which can be used to prepare drugs against the new coronavirus.
[0090] The above description is only a preferred specific embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any technician familiar with the technical field, within the technical scope disclosed by the present invention, can make equivalent replacements or changes based on the technical solutions and inventive concepts of the present invention, which should be covered by the scope of protection of the present invention.
Claims
1. The use of cationic rosin and its cucurbitacin complex in the preparation of drugs for preventing or treating novel coronavirus infection, characterized in that: The molecular formula of the cationic pink is C 22 H 26 Cl2N2; the structural formula of the cationic pink is: The cationic peach red cucurbit ring complex is obtained by reacting a dislocated ten-membered cucurbit ring with cationic peach red; the molecular formula of the cationic peach red complex is: C 80 H 88 N 40 O 20 Cl2N2; the structural formula of the cationic pink complex is:
2. The use of the cationic pink and its complex according to claim 1 in the preparation of a drug for treating anti-new coronavirus infection, characterized in that: The preparation method of the cationic peach red cucurbit ring complex is as follows: 1.5 mg of dislocated ten-membered cucurbit ring and 0.5 mg of cationic peach red are weighed, and the mixture is mixed and reacted to obtain the cationic peach red cucurbit ring complex; the molar ratio of the dislocated ten-membered cucurbit ring to the cationic peach red is 1:
1.
3. The use of the cationic peach red and its cucurbitacin complex according to claim 2 in the preparation of a drug for treating anti-new coronavirus infection, characterized in that: The preparation method of the cationic peach red cucurbitacin complex is as follows: 1.5 mg of the dislocated ten-membered cucurbitacin and 0.5 mg of cationic peach red are weighed, 0.4-2 mL of deionized water are added, and after mixing and reaction, a cationic cucurbitacin pink complex with a concentration of 1-5 mg / mL is obtained; the molar ratio of the dislocated ten-membered cucurbitacin to the cationic peach red is 1:
1.
4. The use of the cationic pink and its cucurbitacin complex according to claim 2 in the preparation of a drug for treating anti-new coronavirus infection, characterized in that: The preparation method of the cationic peach red cucurbitacin complex is as follows: 1.5 mg of the dislocated ten-membered cucurbitacin and 0.5 mg of cationic peach red are weighed, 2 mL of deionized water are added, and after mixing and reaction, a cationic cucurbitacin pink complex with a concentration of 1 mg / mL is obtained; the molar ratio of the dislocated ten-membered cucurbitacin to the cationic peach red is 1:
1.
5. The use of the cationic peach red and the cucurbitacin complex thereof according to any one of claims 1 to 4 in the preparation of a drug for preventing or treating infection with a novel coronavirus, characterized in that: The cationic rosmarin and the cucurbitacin ring complex are combined with acceptable excipients in medicines and processed according to conventional methods to prepare corresponding medicines.
6. The use of the cationic pink and its complex according to claim 5 in the preparation of a drug for preventing or treating infection with the new coronavirus, characterized in that: The medicine is in the form of tablets, capsules, granules and oral liquid.