CN115368300B - Compound for TLR8 inhibitor and preparation method and application thereof - Google Patents
Compound for TLR8 inhibitor and preparation method and application thereof Download PDFInfo
- Publication number
- CN115368300B CN115368300B CN202211322360.4A CN202211322360A CN115368300B CN 115368300 B CN115368300 B CN 115368300B CN 202211322360 A CN202211322360 A CN 202211322360A CN 115368300 B CN115368300 B CN 115368300B
- Authority
- CN
- China
- Prior art keywords
- compound
- tlr8
- methylphenol
- methoxyquinolin
- ints
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D215/00—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems
- C07D215/02—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen atoms or carbon atoms directly attached to the ring nitrogen atom
- C07D215/16—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen atoms or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
- C07D215/20—Oxygen atoms
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H15/00—Compounds containing hydrocarbon or substituted hydrocarbon radicals directly attached to hetero atoms of saccharide radicals
- C07H15/26—Acyclic or carbocyclic radicals, substituted by hetero rings
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Immunology (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Medicinal Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention provides a compound for a TLR8 inhibitor, and a preparation method and application thereof, and relates to the technical field of small molecule drug compounds.
Description
Technical Field
The invention relates to the technical field of small molecule drug compounds, in particular to a compound for a TLR8 inhibitor, and a preparation method and application thereof.
Background
Autoimmune diseases are a series of chronic systemic inflammatory diseases that result in a decrease in tolerance to self-antigens due to a dysregulation of the immune system. Toll-like receptors (TLRs) are a class of important protein molecules involved in nonspecific immunity (innate immunity) and are also bridges for connecting nonspecific immunity and specific immunity.
TLR8 in TLRs family is related to the etiology of some autoimmune diseases aiming at self RNA and DNA/protein complex, so that the inhibitor targeting TLR8 is very likely to become a therapeutic agent of the autoimmune diseases, and the prospect is very wide.
Although the research on the TLR8 target is greatly progressed at present, most products are agonists of TLR8, inhibitors of TLR8 are few, and meanwhile, the existing micromolecule compounds for TLR8 inhibitors are poor in solubility and not ideal in effect.
In view of the above, the present invention is particularly proposed.
Disclosure of Invention
One of the objects of the present invention is to provide a compound for a TLR8 inhibitor, which has advantages of high solubility and strong inhibitory effect on TLR 8.
The invention also aims to provide a preparation method of the compound for the TLR8 inhibitor, which has simple process and easy operation.
The invention also aims to provide application of the compound for the TLR8 inhibitor, which can prepare a medicament with good treatment effect and has outstanding application effect.
In order to achieve the above purpose of the present invention, the following technical solutions are adopted:
in a first aspect, a compound for use in a TLR8 inhibitor has a structure represented by general formula (1), or a pharmaceutically acceptable salt thereof;
wherein R is selected from sulfonic acid group or cyclic glucose group having carboxyl group.
Further, the compound is any one of the following compounds:
In a second aspect, a process for the preparation of a compound according to any one of the preceding claims, comprising the steps of:
the hydroxy hydrogen of 4- (7-methoxyquinolin-4-yl) -2-methylphenol is substituted with a sulfonic acid group or a cyclic glucose group having a carboxyl group to give the compound.
Further, the method for substituting hydroxy hydrogen of 4- (7-methoxyquinolin-4-yl) -2-methylphenol with sulfonic acid group comprises the steps of:
the compound is obtained after the 4- (7-methoxyquinoline-4-yl) -2-methylphenol reacts with pyridine trioxide.
Further, the temperature of the reaction of the 4- (7-methoxyquinoline-4-yl) -2-methylphenol and the sulfur trioxide pyridine is 70 to 90 ℃.
Further, the molar ratio of the 4- (7-methoxyquinoline-4-yl) -2-methylphenol to the sulfur trioxide pyridine is 1:2 to 1:4.
further, the method for substituting hydroxy hydrogen of 4- (7-methoxyquinolin-4-yl) -2-methylphenol with cyclic glucose group having carboxyl group comprises the steps of:
4- (7-methoxyquinoline-4-yl) -2-methylphenol hydrochloride reacts with alpha-D-glucuronic acid methyl ester to obtain a substitution product, and the substitution product is hydrolyzed by ester group and acidified to obtain the compound;
the hydrolysis conditions are alkaline conditions.
Further, 4- (7-methoxyquinolin-4-yl) -2-methylphenol hydrochloride was reacted with alpha-D-glucuronic acid methyl ester at a temperature of 20 to 30 ℃.
Further, the molar ratio of 4- (7-methoxyquinolin-4-yl) -2-methylphenol hydrochloride to alpha-D-glucuronic acid methyl ester was 1:3 to 1:6.
in a third aspect, use of a compound according to any one of the preceding claims in the manufacture of a medicament for the treatment of an autoimmune disease.
Compared with the prior art, the invention has at least the following beneficial effects:
the compound for the TLR8 inhibitor provided by the invention has higher solubility and stronger inhibitory activity to TLR8 through the action of a specific substituent group, and particularly, the structures of a sulfonic group and a glucose group have good water solubility, so that the problems of better rigidity and poorer water solubility of the original substituent group with a benzene ring are solved.
The preparation method of the compound for the TLR8 inhibitor provided by the invention is simple in process and easy to operate.
The application of the compound for the TLR8 inhibitor provided by the invention can prepare a medicament with a good treatment effect, and has an outstanding application effect.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the embodiments or the prior art descriptions will be briefly described below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and other drawings can be obtained by those skilled in the art without creative efforts.
FIG. 1 is a schematic diagram of a synthetic reaction for INTS small molecule compounds according to one embodiment of the present invention;
FIG. 2 is a diagram of a reaction for synthesizing 4- (7-methoxyquinolin-4-yl) -2-methylphenol according to an embodiment of the present invention;
FIG. 3 is a diagram of a synthetic reaction for an INTA small molecule compound according to one embodiment of the present invention;
FIG. 4 is a nuclear magnetic hydrogen spectrum of an INTS small molecule compound provided in example 3 of the present invention;
FIG. 5 is a mass spectrum of an INTS small molecule compound provided in example 3 of the present invention;
FIG. 6 is a nuclear magnetic hydrogen spectrum of an INTA small molecule compound provided in example 4 of the present invention;
FIG. 7 is a mass spectrum of an INTA small molecule compound provided in example 4 of the present invention;
FIG. 8 is a graph showing the inhibitory activity of the INTS small molecule compound obtained in test example 1 of the present invention on HEK-Blue-TLR8 cells;
FIG. 9 is a graph showing the inhibitory activity of the INTA small molecule compound obtained in test example 1 of the present invention on HEK-Blue-TLR8 cells.
Detailed Description
The technical solutions of the present invention will be described clearly and completely with reference to the following embodiments, and it should be understood that the described embodiments are some, but not all, embodiments of the present invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
According to a first aspect of the present invention, there is provided a compound for use in a TLR8 inhibitor, having a structure represented by general formula (1), or a pharmaceutically acceptable salt thereof;
wherein R is selected from sulfonic acid group or cyclic glucose group having carboxyl group.
The compound for the TLR8 inhibitor is connected with a specific substituent group, and has higher solubility and stronger inhibitory activity on TLR8 under the action of the specific substituent group.
In a preferred embodiment, the compound for a TLR8 inhibitor of the invention is any one of the following compounds:
INTS Small molecules INTA Small molecules
The solubility of the INTS small molecular compound provided by the invention under neutral condition is 28.7 mu g/mL, and the inhibitory activity IC of the INTS small molecular compound on TLR8 50 6.31 + -3.11 nM; the solubility of the INTA micromolecule compound provided by the invention under neutral condition is 140 mu g/mL, and the inhibitory activity IC of the INTA micromolecule compound on TLR8 50 168.56. + -. 83.36nM.
According to a second aspect of the present invention there is provided a process for the preparation of a compound as described in any one of the preceding claims, comprising the steps of:
the hydroxy hydrogen of 4- (7-methoxyquinolin-4-yl) -2-methylphenol is substituted with a sulfonic acid group or a cyclic glucose group having a carboxyl group to give the compound.
In the present invention, 4- (7-methoxyquinolin-4-yl) -2-methylphenol (CAS: 2165340-32-7) has the chemical structure:
。
the preparation method of the compound for the TLR8 inhibitor provided by the invention is simple in process and easy to operate.
In a preferred embodiment, the method for substituting the hydroxy hydrogen of 4- (7-methoxyquinolin-4-yl) -2-methylphenol with a sulfonic acid group comprises the steps of:
reacting 4- (7-methoxyquinoline-4-yl) -2-methylphenol with sulfur trioxide pyridine to obtain the compound.
Wherein the chemical structure of sulfur trioxide pyridine (CAS: 26412-87-3) is as follows:
。
in a preferred embodiment, the temperature for the reaction of 4- (7-methoxyquinolin-4-yl) -2-methylphenol and pyridine trioxide is from 70 ℃ to 90 ℃, for example, 70 ℃, 75 ℃, 80 ℃, 85 ℃ and 90 ℃, but not limited thereto, and preferably 80 ℃, which is more favorable for the reaction and improves the reaction effect.
In a preferred embodiment, the molar ratio of 4- (7-methoxyquinolin-4-yl) -2-methylphenol to sulfur trioxide pyridine is 1: 3. 1:4, but not limited thereto, may preferably be 1:3, the reaction is more favorably carried out, and the reaction effect is improved.
A typical preparation method of an INTS small molecule compound, see fig. 1, comprises the following steps:
4- (7-Methoxyquinolin-4-yl) -2-methylphenol (7.9g, 30 mmol) was dissolved in 100 mL of DMF to prepare a solution, and then pyridine trioxide (14.3g, 90 mmol) was added to obtain a mixture;
the resulting mixture is then stirred overnight at 80 ℃ (overnight) and upon completion the resulting product precipitates and can be filtered and washed with DMF to give a pure yellow solid (containing DMF) which can be completely removed after drying in vacuo at 130 ℃ for about 30h to give the dried INTS small molecule compound (about 6.0g, about 60% yield).
In the present invention, 4- (7-methoxyquinolin-4-yl) -2-methylphenol may be obtained by commercially available methods; alternatively, it can be prepared by reacting 4- (7-methoxyquinolin-4-yl) -2-methylphenolate, according to the reaction scheme shown in FIG. 2, comprising the steps of:
to 150 mL of DMF was added 4- (7-methoxyquinolin-4-yl) -2-methylphenolate (15.1g, 50 mmol) and stirred at room temperature to form a suspension, giving a mixture;
then K is put 2 CO 3 (7g, 50 mmol) of 50 mL of an aqueous solution was slowly added to the mixture and stirred at room temperature (rt) for 4 hours, after completion of the reaction, the mixture was extracted with EA (ethyl acetate) 3 times with 200 mL of EA to give an organic layer containing the product, which was then further extracted with H 2 O washing, then adding Na to the organic layer 2 SO 4 And dried, and the solvent was removed in vacuo to give 4- (7-methoxyquinolin-4-yl) -2-methylphenol (about 12g, yield about 91%).
The preparation method of the INTS micromolecule compound provided by the invention is simple and efficient in process and easy to operate.
In a preferred embodiment, the method of the present invention for substituting the hydroxy hydrogen of 4- (7-methoxyquinolin-4-yl) -2-methylphenol with a cyclic glucose group having a carboxyl group comprises the steps of:
reacting 4- (7-methoxyquinoline-4-yl) -2-methylphenol hydrochloride with alpha-D-glucuronic acid methyl ester to obtain a substitution product, hydrolyzing an ester group, and acidifying to obtain a compound;
wherein, the condition of hydrolysis can be alkaline condition.
The chemical structure of alpha-D-glucuronic acid methyl ester (CAS: 21085-72-3) of the invention is:
。
in a preferred embodiment, the temperature for reacting 4- (7-methoxyquinolin-4-yl) -2-methylphenol hydrochloride with a-D-glucuronic acid methyl ester is 20 to 30 ℃, for example, 20 ℃, 25 ℃ or 30 ℃, but not limited thereto, and preferably 25 ℃, which is more favorable for the reaction and improves the reaction effect.
In a preferred embodiment, the molar ratio of 4- (7-methoxyquinolin-4-yl) -2-methylphenol hydrochloride to a-D-glucuronic acid methyl ester is 1:3 to 1:6, for example, may be 1: 3. 1:4. 1: 5. 1:6, may preferably be 1:5, but not limited to, the method is more favorable for the reaction and improves the reaction effect.
A typical preparation method of an INTA small molecule compound, see fig. 3, comprises the following steps:
4- (7-Methoxyquinolin-4-yl) -2-methylphenolate hydrochloride (40.0 g,136 mmol, 1eq) was degassed in 200 mL of quinoline (quinoline) and N was used to degas 2 Washing 3 times, then N 2 Stirring at 25 ℃ for 25 minutes under an atmosphere, and then adding Ag 2 O (184g, 795 mmol, 6.00eq) and molecular sieve 4A (200g, 199 mmol, 1.50eq) and stirred at 0 ℃ for 20 minutes to give a mixture;
then methyl alpha-D-glucuronide (263g, 663 mmol, 5.00eq) was added to the above mixture at N 2 After completion of the reaction, the product was obtained by filtration through celite under stirring at 25 ℃ for 2 hours, and the solvent was removed in vacuo, and then purified by column chromatography using a mixture of petroleum ether and ethyl acetate as a developing solvent (the volume ratio of the two may be 1, but is not limited thereto) to obtain an intermediate (about 35.5g,61.0mmol, yield about 46.0%) as a brown solid;
the intermediate (30.0 g,51.6 mmol, 1eq) was dissolved in 300 mL of methanol (MeOH) to form a solution, and lithium hydroxide monohydrate (LiOH. H) was added 2 O) (21.7 g,516 mmol,10.0 eq) and water (10.0 mL) to give a mixture, which is stirred at 20-25 ℃ for 1 hour, then HCOOH is added to adjust the pH to 5-6 to give the crude product, which is purified by reverse phase HPLC (0.1% FA conditions) to give a fraction with MS of 442.2, which is frozen to give a crude product containing INTA, which is then purified by reverse phase HPLC (0.1% FA conditions) and concentrated to give the finished INTA product (7.82g, 17.5 mmol, yield about 21.7% as an off-white solid.
The preparation method of the INTA micromolecule compound provided by the invention has the advantages of simple and efficient process and easiness in operation.
According to a third aspect of the present invention there is provided the use of a compound as described in any one of the preceding aspects in the manufacture of a medicament for the treatment of an autoimmune disease.
The application of the compound for the TLR8 inhibitor provided by the invention can prepare a medicament with a good treatment effect, and has an outstanding application effect.
The invention is further illustrated by the following examples. The materials in the examples were prepared according to the existing methods or directly commercially available, unless otherwise specified.
Example 1
A compound useful as a TLR8 inhibitor (denoted as INTS) having the structure:
example 2
A compound for use in a TLR8 inhibitor (denoted as INTA) having the structure:
example 3
This example is a process for the preparation of the INTS small molecule compound of example 1, comprising the steps of:
s1: to 150 mL of DMF was added 4- (7-methoxyquinolin-4-yl) -2-methylphenolate (15.1g, 50 mmol) and stirred at room temperature to form a suspension, giving a mixture;
then K is added 2 CO 3 (7g, 50 mmol) of 50 mL of an aqueous solution was slowly added to the above mixture and stirred at room temperature for 4 hours, after completion of the reaction, the mixture was extracted with EA (ethyl acetate) 3 times with 200 mL each to give an organic layer containing the product, which was further extracted with H 2 O washing, then adding Na to the organic layer 2 SO 4 And dried, and the solvent removed in vacuo to give 4- (7-methoxyquinolin-4-yl) -2-methylphenol (about 12g, yield about 91%);
s2:4- (7-Methoxyquinolin-4-yl) -2-methylphenol (7.9g, 30 mmol) was dissolved in 100 mL of DMF to prepare a solution, and then pyridine trioxide (14.3g, 90 mmol) was added to obtain a mixture;
the resulting mixture is then stirred at 80 ℃ overnight (overlap), after completion of the reaction the product obtained is a precipitate which can be filtered and washed with DMF to give a pure yellow solid (containing DMF) which can be completely removed after drying in vacuo at 130 ℃ for about 30h to give the dried INTS small molecule compound (about 6.0g, yield about 60%);
the NMR spectrum of the INTS small molecule compound obtained in this example is shown in FIG. 4, and the test conditions are 400 MHz, DMSO-d 6 ， 1 H NMR (400 MHz, DMSO-d 6 ) :δ 9.17 (d, J = 6.0 Hz, 1H, ArH-1), 8.13 (d, J =10.0 Hz, 1H, ArH-6), 7.82 (d, J = 6.0 Hz, 1H, ArH-2), 7.64 (d, J = 8.4 Hz, 1H, ArH-3), 7.58-7.55 (m, 2H, ArH-7/8), 7.48 (d, J = 2.4 Hz, 1H, ArH-5), 7.44 (dd, J =2.4, 8.4 Hz, 1H, ArH-4), 4.04 (s, 3H, OCH 3 -9), 2.31 (s, 3H, CH 3 -10); the mass spectrum of the INTS small molecule compound is shown in figure 5, and the mass spectrum testing conditions are as follows: TOF-MS-ESI +, solvent: meCN/H 2 O; the nuclear magnetic hydrogen spectrum and the mass spectrum jointly prove the correct structure of the INTS small molecular compound.
Example 4
This example is a process for the preparation of the INTA small molecule compound of example 2, comprising the steps of:
s1: 4- (7-Methoxyquinolin-4-yl) -2-methylphenolate hydrochloride (40.0 g,136 mmol, 1eq) was degassed in 200 mL of quinoline (quinoline) and N was used to degas 2 Washing 3 times, then N 2 Stirring at 25 ℃ for 25 minutes under an atmosphere, and then adding Ag 2 O (184g, 795 mmol, 6.00eq) and molecular sieve 4A (200g, 199 mmol, 1.50eq) and stirred at 0 ℃ for 20 minutes to give a mixture;
then alpha-D-glucuronic acid methyl ester (263g, 663 mmol, 5.00eq) was added to the above mixture, at N 2 The product was obtained by filtration through celite after completion of the reaction under stirring at 25 ℃ for 2 hours under an atmosphere, and the solvent was removed in vacuo, followed by purification by column chromatography using a mixture of petroleum ether and ethyl acetate as a developing solvent (the volume ratio of the two may be 1).
The intermediate (30.0 g,51.6 mmol, 1eq) was dissolved in 300 mL of methanol (MeOH) to form a solution, and lithium hydroxide monohydrate (LiOH. H) was added 2 O) (21.7 g,516 mmol,10.0 eq) and water (10.0 mL) to give a mixture, which is stirred at 20-25 ℃ for 1 hour, then HCOOH is added to adjust the pH to 5-6 to give the crude product, which is purified by reverse phase HPLC (0.1% FA conditions) to give a fraction with MS of 442.2, which is frozen to give a crude product containing INTA, which is then purified by reverse phase HPLC (0.1% FA conditions) and concentrated to give the finished INTA product (7.82g, 17.5 mmol, yield about 21.7% as an off-white solid;
the nuclear magnetic hydrogen spectrum of the INTA small molecular compound obtained in the example is shown in FIG. 6, and the test conditions are 400 MHz and DMSO-d 6 ， 1 H NMR:(400 MHz, DMSO-d 6 ) ：δ 8.82 (d, J = 5.2 Hz, 1H, ArH-1), 7.81 (d, J =9.2 Hz, 1H, ArH-6), 7.46 (d, J = 5.2 Hz, 1H, ArH-2), 7.34-7.30 (m, 2H, ArH-3/7), 7.27-7.20 (m, 3H, ArH-4/5/8), 5.11 (d, J = 7.2 Hz, 1H, H-1'), 3.48-3.33 (m, 4H, H-2'/3'/4'/5'), 3.93 (s, 3H, OCH 3 -9), 2.21 (s, 3H, CH 3 -10); the mass spectrum of the INTA small molecule compound is shown in figure 7, and the mass spectrum testing conditions are as follows: ion mode ESI, solvent MeCN (0.01% TFA); the nuclear magnetic hydrogen spectrum and the mass spectrum jointly prove the correct structure of the INTA small molecular compound in the embodiment.
Comparative example 1
A small molecule compound inhibitor of TLR8, having the structure:
the Small Molecule compound inhibitors of this comparative example are derived from the literature of the prior art (Hu, Z., et al., small-molecular TLR8 antibodies via Structure-Based Rational design. Cell Chem Biol, 2018.25 (10): 1286-1291 e 1283), IC, for their inhibitory activity against TLR8 50 The concentration was 15.17. + -. 5.38uM.
Test example 1
The activity of the INTS small molecule compound provided in example 1 in HEK-Blue-TLR8 cells was tested by measuring the activity of the INTS small molecule compound in HEK-Blue-TLR8 cells using the SEAP Assay method;
the plate paving mode is as follows:
| 0.0152 | 0.0457 | 0.1371 | 0.4115 | 1.2345 | 3.7037 | 11.1111 | 33.3333 | 100 C ell only |
| 0 | 0 | 6.96917 E- 06 | 2.09075 E-05 | 6.27225E-05 | 0.00019 | 0.00056 | 0.0017 | 0.0051 DMEM only |
wherein, the Cell only wells contain TLR8 cells and FBS-free DMEM medium, but do not contain INTS drugs and R848 activator; DMEM only wells contained DMEM medium without FBS and no TLR8 cells, INTS drug and R848 activator; the rest holes are DMEM culture medium which is simultaneously added with TLR8 cells, INTS medicines, R848 activator and FBS-free, and the concentration unit of the INTS medicines is mu m/mL; e represents scientific counting method;
1. preparing INTS medicine in a new 96-well plate by DMEM without FBS in a 3-fold dilution gradient, so that the concentration of the medicine in the corresponding well is 3 times of the target concentration;
2. absorbing the original culture medium in the HEK Blue hTLR8 cell culture dish, blowing and beating the culture medium by using DMEM without FBS, and counting;
3. transferring the prepared INTS drug to a new 96-well plate, wherein each well is 50 mu L, and then adding 50 mu L of cells with or without R848, wherein the concentration of R848 is 2 mu g/mL, so that the final concentration of the activator in the plate is 1 mu g/mL, and the number of cells in each well is about 2 ten thousand per well;
4. after culturing in an incubator for 24 h, taking 50 mu L of supernatant to a new 96-well plate, adding 50 mu L of Quanti-Blue, and testing the absorbance at 620 nm after the negative control well (the cell well only added with R848) turns Blue;
5. data were obtained and analyzed, see fig. 8.
The result is that the INTS small molecular compound has activity IC on TLR8 cells 50 6.31. + -. 3.11nM.
Note: the TLR8 is a Toll-like receptor 8, is an important pathogenic mode recognition receptor of an immune system, can recognize ssRNA of virus so as to activate the immune system of an organism, and the HEK-Blue-TLR8 cell overexpresses TLR8 in a HEK293T cell and is added with an SEAP report system, so that the change of downstream signals of the TLR8 can be read by using an instrument, and the regulation and control effect of an added INTS small molecular compound on the TLR8 can be further represented.
The small INTA compounds provided in example 2 were tested in the same manner as described above, and the data were obtained and analyzed, as shown in FIG. 9, with the result that IC, the activity of small INTA compounds on TLR8 cells, was IC 50 168.56. + -. 83.36nM.
Thus, the inhibitory activity IC of the INTS small molecule compound provided by the invention on TLR8 50 6.31 +/-3.11nM, and the inhibitory activity IC of INTA small molecular compound on TLR8 50 The activity of the compound is 168.56 +/-83.36 nM, which is obviously superior to the activity of the small molecule compound reported in the literature in comparative example 1 on the inhibition of TLR8, and the small molecule compound provided by the application has strong inhibition effect on TLR 8.
Test example 2
The solubility of the INTS small molecule compound provided in example 1, the INTA small molecule compound provided in example 2 and the small molecule compound provided in comparative example 1 were tested as follows:
physiological saline is used as a buffer solution;
the experimental steps are as follows:
(1) Adding the powder of the substance to be tested into normal saline according to the proportion of 1;
(2) Fully oscillating for 4 hours;
(3) Centrifuging (centrifugal force 15700 g, centrifuging time 20 min), taking the supernatant, diluting 2000 times with acetonitrile, mixing uniformly, injecting and analyzing, and obtaining the following results:
note: NA means pH is neutral
Finally, it should be noted that: the above embodiments are only used to illustrate the technical solution of the present invention, and not to limit the same; while the invention has been described in detail and with reference to the foregoing embodiments, it will be understood by those skilled in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some or all of the technical features may be equivalently replaced; and these modifications or substitutions do not depart from the spirit of the corresponding technical solutions of the embodiments of the present invention.
Claims (7)
1. A compound for use as a TLR8 inhibitor, having a structure represented by general formula (1), or a pharmaceutically acceptable salt thereof;
wherein R is selected from cyclic glucose groups having a carboxyl group.
2. The compound of claim 1, wherein the compound is the following:
3. a process for the preparation of a compound according to claim 1 or 2, comprising the steps of:
the compound is obtained after the hydroxy hydrogen of 4- (7-methoxyquinolin-4-yl) -2-methylphenol is substituted with a cyclic glucose group having a carboxyl group.
4. The production method according to claim 3, wherein the method of substituting hydroxy hydrogen of 4- (7-methoxyquinolin-4-yl) -2-methylphenol with a cyclic glucose group having a carboxyl group comprises the steps of:
4- (7-methoxyquinoline-4-yl) -2-methylphenol hydrochloride reacts with alpha-D-glucuronic acid methyl ester to obtain a substitution product, and the substitution product is hydrolyzed by ester group and acidified to obtain the compound;
the hydrolysis conditions are alkaline conditions.
5. The process according to claim 4, wherein 4- (7-methoxyquinolin-4-yl) -2-methylphenol hydrochloride is reacted with A-D-glucuronic acid methyl ester at a temperature of 20 to 30 ℃.
6. The process according to claim 4, wherein the molar ratio of 4- (7-methoxyquinolin-4-yl) -2-methylphenol hydrochloride to A-D-glucuronic acid methyl ester is 1:3 to 1:6.
7. use of a compound according to claim 1 or 2 in the manufacture of a medicament for the treatment of an autoimmune disease.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211322360.4A CN115368300B (en) | 2022-10-27 | 2022-10-27 | Compound for TLR8 inhibitor and preparation method and application thereof |
| PCT/CN2023/104858 WO2024087720A1 (en) | 2022-10-27 | 2023-06-30 | Compound for tlr8 inhibitor, and preparation method therefor and use thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211322360.4A CN115368300B (en) | 2022-10-27 | 2022-10-27 | Compound for TLR8 inhibitor and preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN115368300A CN115368300A (en) | 2022-11-22 |
| CN115368300B true CN115368300B (en) | 2023-03-24 |
Family
ID=84072982
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202211322360.4A Active CN115368300B (en) | 2022-10-27 | 2022-10-27 | Compound for TLR8 inhibitor and preparation method and application thereof |
Country Status (2)
| Country | Link |
|---|---|
| CN (1) | CN115368300B (en) |
| WO (1) | WO2024087720A1 (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115368300B (en) * | 2022-10-27 | 2023-03-24 | 北京拓领博泰生物科技有限公司 | Compound for TLR8 inhibitor and preparation method and application thereof |
Family Cites Families (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1223115A (en) * | 1992-04-24 | 1999-07-21 | 武田药品工业株式会社 | Pharmaceutical composition containing quinoline and quinazoline derivatives and novel compounds therefor |
| JP2007524615A (en) * | 2003-06-20 | 2007-08-30 | コーリー ファーマシューティカル ゲーエムベーハー | Low molecular weight Toll-like receptor (TLR) antagonist |
| EP2890808A4 (en) * | 2012-08-29 | 2016-09-28 | California Inst Of Techn | Diagnosis and treatment of autism spectrum disorder |
| CN104490617B (en) * | 2014-11-24 | 2017-06-16 | 佛山市金骏康健康科技有限公司 | Dihydrochalcone glucosides is used as whitening, the purposes of the dark and gloomy skin quality active ingredient of improvement |
| CN117659025A (en) * | 2017-10-30 | 2024-03-08 | 科罗拉多州立大学董事会法人团体 | TOLL-like receptor 8 (TLR 8) specific antagonists, methods of making and uses thereof |
| US20220363665A1 (en) * | 2019-09-10 | 2022-11-17 | Hoffmann-La Roche Inc. | Quinoline compounds for the treatment of autoimmune disease |
| CN114478374B (en) * | 2022-04-07 | 2022-07-15 | 北京拓领博泰生物科技有限公司 | Toll-like receptor 8 specific inhibitor hydrochloride and preparation method and application thereof |
| CN114426518B (en) * | 2022-04-07 | 2022-07-15 | 北京拓领博泰生物科技有限公司 | Phosphate salts of TOLL-like receptor inhibiting compounds and methods for preparing same |
| CN114591235A (en) * | 2022-04-07 | 2022-06-07 | 北京拓领博泰生物科技有限公司 | Pharmaceutically acceptable salt of 4- (7-methoxyquinoline-4-yl) -2-methylphenol and preparation method thereof |
| CN115368300B (en) * | 2022-10-27 | 2023-03-24 | 北京拓领博泰生物科技有限公司 | Compound for TLR8 inhibitor and preparation method and application thereof |
-
2022
- 2022-10-27 CN CN202211322360.4A patent/CN115368300B/en active Active
-
2023
- 2023-06-30 WO PCT/CN2023/104858 patent/WO2024087720A1/en unknown
Also Published As
| Publication number | Publication date |
|---|---|
| CN115368300A (en) | 2022-11-22 |
| WO2024087720A1 (en) | 2024-05-02 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111961057A (en) | Alpha-configuration nucleoside and application thereof in treating feline coronavirus infection | |
| CN110092745B (en) | Compound containing aromatic ring and application thereof | |
| CN115368300B (en) | Compound for TLR8 inhibitor and preparation method and application thereof | |
| CN113278041B (en) | Nucleoside phosphate and its synthesis method and application in preparing medicine for anti hepatitis virus | |
| WO2022174525A1 (en) | Compound, preparation method therefor and use thereof | |
| CN112939824A (en) | Compound and medical application thereof in colorectal cancer | |
| CN111196801B (en) | Aporphine alkaloid derivative and preparation method and application thereof | |
| CN114315933B (en) | Preparation method of potential anti-new coronavirus drug monatin | |
| CN115353508A (en) | 5-pyridine-1H-indazole compound, pharmaceutical composition and application | |
| CN109970679B (en) | Paeonol thiazole derivative and preparation method and application thereof | |
| CN113416189B (en) | Beta-carbopol Lin Dangai derivative, preparation method and anti-tumor application | |
| CN113149942A (en) | Rockmilanol phenolic hydroxyl derivative, preparation method and application thereof | |
| CN115057831B (en) | Thiadiazole compound and application thereof in preparation of KKBC 1 protein inhibitor | |
| CN105646546A (en) | Acid-sensitive camptothecin-20-position ester derivative and antineoplastic application thereof | |
| CN110563795B (en) | Preparation method and application of diosgenin derivative containing 1,3,4 oxadiazole or 1,3,4 thiadiazole fragment | |
| CN111848629B (en) | mTOR/HDAC dual inhibitor and application thereof | |
| CN114560845B (en) | Crystal form alpha of quinoline compound, and preparation method and application thereof | |
| CN114890928B (en) | Isothiocyanate derivative and preparation method and application thereof | |
| CN113563330B (en) | 3-position derivative of beta-carbopol as well as preparation method and application thereof | |
| CN115340502B (en) | BCL-XL inhibitor and preparation method and application thereof | |
| CN109369642B (en) | Related substance of a kind of Eliquis and its preparation method and application | |
| CN117551082A (en) | 2-arylthiomethyl-6-bromoindole compound and synthesis and application thereof | |
| CN107698511B (en) | 1, 3-diaryl-5-alkoxy pyrazole compound and preparation method and application thereof | |
| CN106831925B (en) | A kind of cholesterol-Biotin molecular probe and its preparation method and application | |
| CN117343069A (en) | Novel compound, pharmaceutical composition and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |