CN111574334A - Novel phenolic compound and preparation method and application thereof - Google Patents
Novel phenolic compound and preparation method and application thereof Download PDFInfo
- Publication number
- CN111574334A CN111574334A CN202010298193.9A CN202010298193A CN111574334A CN 111574334 A CN111574334 A CN 111574334A CN 202010298193 A CN202010298193 A CN 202010298193A CN 111574334 A CN111574334 A CN 111574334A
- Authority
- CN
- China
- Prior art keywords
- compound
- reacting
- novel phenolic
- benzyl
- phenolic compound
- 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.)
- Granted
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C39/00—Compounds having at least one hydroxy or O-metal group bound to a carbon atom of a six-membered aromatic ring
- C07C39/24—Halogenated derivatives
- C07C39/367—Halogenated derivatives polycyclic non-condensed, containing only six-membered aromatic rings as cyclic parts, e.g. halogenated poly-hydroxyphenylalkanes
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N31/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic oxygen or sulfur compounds
- A01N31/08—Oxygen or sulfur directly attached to an aromatic ring system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/045—Hydroxy compounds, e.g. alcohols; Salts thereof, e.g. alcoholates
- A61K31/05—Phenols
- A61K31/055—Phenols the aromatic ring being substituted by halogen
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C37/00—Preparation of compounds having hydroxy or O-metal groups bound to a carbon atom of a six-membered aromatic ring
- C07C37/01—Preparation of compounds having hydroxy or O-metal groups bound to a carbon atom of a six-membered aromatic ring by replacing functional groups bound to a six-membered aromatic ring by hydroxy groups, e.g. by hydrolysis
- C07C37/055—Preparation of compounds having hydroxy or O-metal groups bound to a carbon atom of a six-membered aromatic ring by replacing functional groups bound to a six-membered aromatic ring by hydroxy groups, e.g. by hydrolysis the substituted group being bound to oxygen, e.g. ether group
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C37/00—Preparation of compounds having hydroxy or O-metal groups bound to a carbon atom of a six-membered aromatic ring
- C07C37/11—Preparation of compounds having hydroxy or O-metal groups bound to a carbon atom of a six-membered aromatic ring by reactions increasing the number of carbon atoms
- C07C37/20—Preparation of compounds having hydroxy or O-metal groups bound to a carbon atom of a six-membered aromatic ring by reactions increasing the number of carbon atoms using aldehydes or ketones
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C37/00—Preparation of compounds having hydroxy or O-metal groups bound to a carbon atom of a six-membered aromatic ring
- C07C37/62—Preparation of compounds having hydroxy or O-metal groups bound to a carbon atom of a six-membered aromatic ring by introduction of halogen; by substitution of halogen atoms by other halogen atoms
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C41/00—Preparation of ethers; Preparation of compounds having groups, groups or groups
- C07C41/01—Preparation of ethers
- C07C41/16—Preparation of ethers by reaction of esters of mineral or organic acids with hydroxy or O-metal groups
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C41/00—Preparation of ethers; Preparation of compounds having groups, groups or groups
- C07C41/01—Preparation of ethers
- C07C41/18—Preparation of ethers by reactions not forming ether-oxygen bonds
- C07C41/30—Preparation of ethers by reactions not forming ether-oxygen bonds by increasing the number of carbon atoms, e.g. by oligomerisation
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C67/00—Preparation of carboxylic acid esters
- C07C67/14—Preparation of carboxylic acid esters from carboxylic acid halides
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Medicinal Chemistry (AREA)
- Wood Science & Technology (AREA)
- Engineering & Computer Science (AREA)
- Pest Control & Pesticides (AREA)
- Environmental Sciences (AREA)
- Zoology (AREA)
- Epidemiology (AREA)
- Plant Pathology (AREA)
- Agronomy & Crop Science (AREA)
- Oncology (AREA)
- Communicable Diseases (AREA)
- Dentistry (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
Abstract
The invention provides a novel phenolic compound and a preparation method and application thereof, wherein the chemical general formula of the novel phenolic compound is shown as a formula (1), wherein R is Br, Cl or F. The novel phenolic compound provided by the invention can obviously inhibit the growth of gram-positive bacteria, has the minimum inhibitory concentration of 1-4 mu g/mL on staphylococcus aureus, bacillus and methicillin-resistant staphylococcus aureus, does not generate drug resistance under long-time low-dose exposure, and can be used as a gram-positive bacteriumThe antibacterial agent of the 'Shih' positive bacteria has good antibacterial effect.
Description
Technical Field
The invention relates to the technical field of preparation of phenolic compounds, in particular to a novel phenolic compound and a preparation method and application thereof.
Background
Bacterial infections are increasingly becoming a public health hazard, and the abuse of antibiotics has led to the emergence of resistance in a large number of bacteria. Among the gram-positive bacteria, the most important drug-resistant bacteria are methicillin-resistant staphylococcus aureus, which seriously affect the physical health of human beings. The antibacterial agent is a core component of the antibacterial material, and has a function of killing or inhibiting microorganisms. Patent application CN106667994A discloses the application of oligomeric polyphenol compounds in the preparation of anti-gram-positive drug-resistant bacteria products, and the oligomeric polyphenol compounds are dimer and trimer oligomeric polyphenol compounds with trans-structures, and the dimer and trimer resveratrol oligomeric polyphenol compounds are compound 1: trans-D-viniferin and compound II: and the dimer drug-resistant bacterium resistance is stronger than that of the trimer, the MIC value of the dimer trans-D-viniferin is 8 mug/mL, and the MIC value of the trimer gentian H is 16 mug/mL. However, the MIC of these two oligomeric polyphenolic compounds was still high.
Therefore, how to prepare a product with better antibacterial effect against gram-positive drug-resistant bacteria becomes a technical problem to be solved urgently.
Disclosure of Invention
The invention aims to provide a novel phenolic compound, a preparation method and an application thereof, wherein the novel phenolic compound can obviously inhibit the growth of gram-positive bacteria, and the minimum inhibitory concentration to staphylococcus aureus, bacillus subtilis and methicillin-resistant staphylococcus aureus is 1-4 mug/mL.
In order to achieve the above objects, one of the objects of the present invention is to provide a novel phenolic compound having a chemical formula:
wherein, R ═ Br, Cl or F.
Another object of the present invention is to provide a process for producing a novel phenol compound of formula (1), wherein when R ═ Br or Cl is contained in the novel phenol compound, the process comprises:
reacting 4-hydroxydiphenylmethane with a halogen-containing compound to obtain a compound II, wherein the halogen comprises bromine or chlorine;
reacting the compound II with acetyl chloride to obtain a compound III;
reacting the compound III with a phenol solution to obtain the novel phenolic compound;
wherein the chemical general formula of the compound II is the following formula (2); the chemical general formula of the compound III is shown as a formula (3);
in the formulae (2) and (3), R1 ═ Br or Cl.
Further, the reacting 4-hydroxydiphenylmethane with a halogen-containing compound comprises:
dissolving 4-hydroxy diphenylmethane and a halogen-containing compound in an acetic acid or methanol solvent, and reacting at the temperature of 5-25 ℃.
Further, said reacting said compound II with acetyl chloride comprises:
dissolving the compound II and acetyl chloride in a dichloromethane solvent, and reacting in a catalyst AlCl3And reacting at the temperature of 5-25 ℃.
Further, the reacting the compound III with a phenol solution comprises:
and dissolving the compound III in a phenol solution, adding toluenesulfonic acid at the temperature of 25-65 ℃, and stirring for reaction.
The present invention also provides a process for producing a novel phenolic compound of formula (1), wherein when R ═ F in the novel phenolic compound, the process comprises:
reacting 4-Br-2, 6-difluorophenol with bromotoluene to obtain 5-Br-1, 3-difluoro-2-phenyl-methoxybenzene;
reacting the 5-Br-1, 3-difluoro-2-phenyl-methoxybenzene with benzyl magnesium chloride to obtain 4-benzyl-2, 6-dichloro-methoxybenzene;
carrying out reduction reaction on the 4-benzyl-2, 6-dichloro-methoxybenzene to obtain 4-benzyl-2, 6-difluorophenol;
reacting the 4-benzyl-2, 6-difluorophenol with acetyl chloride to obtain 4- (4-acetyl benzyl) -2, 6-difluorophenylacetic acid;
and (3) reacting the 4- (4-acetyl benzyl) -2, 6-difluorophenylacetic acid with a phenol solution to obtain the novel phenolic compound.
Further, the reaction of 4-Br-2, 6-difluorophenol with bromotoluene comprises:
4-Br-2, 6-difluorophenol was dissolved in DMF solution and washed with N2Adding alkaline substances into the mixture under the atmosphere, stirring the mixture, and adding bromotoluene into the mixture after stirring the mixture to react.
Further, said reacting said 5-Br-1, 3-difluoro-2-phenyl-methoxybenzene with benzylmagnesium chloride comprises:
reacting ZnBr2Mixing with THF to obtain a first mixed solution;
in N2And mixing and stirring the benzyl magnesium chloride and the first mixed solution under the atmosphere, and adding 5-Br-1, 3-difluoro-2-phenyl-methoxybenzene to react after stirring.
The fourth purpose of the invention is to provide the application of the novel phenolic compound in preparing gram-positive bacteria antibacterial agent.
Further, the gram-positive bacteria include staphylococcus aureus, bacillus subtilis and methicillin-resistant staphylococcus aureus.
One or more technical solutions in the embodiments of the present invention have at least the following technical effects or advantages:
the novel phenolic compound provided by the invention can obviously inhibit the growth of gram-positive bacteria, has the minimum inhibitory concentration of 1-4 mu g/mL on staphylococcus aureus, bacillus subtilis and methicillin-resistant staphylococcus aureus, does not generate drug resistance under long-time low-dose exposure, can be used as a gram-positive bacteria antibacterial agent, has a good antibacterial effect, and is a good antibacterial agent.
Drawings
In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the drawings needed to be used in the description of the embodiments are briefly introduced below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and it is obvious for those skilled in the art to derive other drawings without creative efforts.
FIG. 1 is a mass spectrum of a novel phenolic compound prepared in example 1 of the present invention;
FIG. 2 is a mass spectrum of a novel phenolic compound prepared in example 2 of the present invention;
FIG. 3 is a mass spectrum of the novel phenolic compound prepared in example 3 of the present invention.
Detailed Description
The present invention will be described in detail below with reference to specific embodiments and examples, and the advantages and various effects of the present invention will be more clearly apparent therefrom. It will be understood by those skilled in the art that these specific embodiments and examples are for the purpose of illustrating the invention and are not to be construed as limiting the invention.
Throughout the specification, unless otherwise specifically noted, terms used herein should be understood as having meanings as commonly used in the art. Accordingly, unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. If there is a conflict, the present specification will control.
Unless otherwise specifically stated, various raw materials, reagents, instruments, equipment and the like used in the present invention are commercially available or can be prepared by existing methods.
In order to solve the technical problems, the embodiment of the invention provides the following general ideas:
to achieve the above objectives, this example provides a novel phenolic compound, which has the chemical formula:
wherein, R ═ Br, Cl or F.
The embodiment of the invention also provides a preparation method of the novel phenolic compound, which comprises the following steps:
1. when R ═ Br or Cl in the novel phenolic compound of formula (1), the preparation method comprises:
step 1, reacting 4-hydroxy diphenylmethane with a halogen-containing compound to obtain a compound II, wherein the halogen comprises bromine or chlorine; in particular, the amount of the solvent to be used,
(1) when R is Br, 4-hydroxy diphenylmethane is dissolved in acetic acid solution, then bromine gas can be directly introduced for reaction, the halogen-containing compound is bromine gas, and the preferred range of the molar ratio of 4-hydroxy diphenylmethane to bromine gas is 15-17: 32 to 34, and can fully react within the molar ratio range;
preferably, the reaction is also carried out with the addition of an organic solvent such as chloroform, acetic acid, methanol, etc., and acetic acid, AcOH, is preferred in this example because it was experimentally found that acetic acid yield is highest for this structure.
(2) When R is Cl, the reaction efficiency is low due to the introduction of chlorine, 4-hydroxy diphenylmethane is dissolved in methanol solution, and then a mixture of NaOH, NaCl and NaClO is added as the halogen-containing compound,
the mol ratio of NaOH to NaCl to NaClO is preferably 2-3: 4.5-5: 4.5-5; the reason for this choice of the molar ratios of NaOH, NaCl and NaClO is that: the applicant first chooses a molar ratio of 1.5: 3: 3, the raw materials are found to be remained, and the reason analysis shows that the effective content of NaClO is probably insufficient, so that the screening is carried out, and the equivalent of NaClO is found to be increased to complete the reaction. Therefore, the application discovers that the molar ratio of NaOH, NaCl and NaClO is 2-3: 4.5-5: the range of 4.5 to 5 enables the reaction to be complete.
Preferably, the reaction is also fed with an organic solvent, such as chloroform, acetic acid, methanol, etc., methanol MeOH is preferred in this example because it was found experimentally that the yield is highest with methanol for this structure.
Step 2, reacting the compound II with acetyl chloride to obtain a compound III;
preferably, the reaction is carried out in a dichloromethane solvent, and a catalyst AlCl is added3The reaction temperature is 5-25 ℃;
the reaction is typically a friedel-crafts acylation reaction, for which: the solvent for the reaction is most commonly dichloromethane, dichloroethane, carbon disulfide and the like; the catalyst for the reaction is most commonly aluminum trichloride, anhydrous zinc chloride, ferric trichloride, titanium tetrachloride and the like; the reaction selects dichloromethane as solvent, AlCl3The catalyst is completely because the two substances are cheap and easy to obtain, and the yield is also high, so that the catalyst is suitable for amplification;
acetyl chloride is added firstly to protect phenolic hydroxyl; in the process, the temperature is hardly influenced between 5 and 25 ℃, but the acetyl protecting the phenolic hydroxyl can be dropped due to the overhigh temperature, so that AlCl is added later3When Friedel-crafts acylation reaction is carried out, side reaction can occur due to the strong positioning effect of phenolic hydroxyl; adding AlCl3In the process, the temperature is not required to be too high (more preferably 5-15 ℃), so that firstly, the reaction is prevented from being too violent, and secondly, side reactions caused by local too high temperature in the system are worried about; and after the addition is finished, naturally heating to room temperature.
Preferably, the molar ratio of the compound II to acetyl chloride is preferably in the range of 1-3: 4-6, the reaction can be completed.
Step 3, reacting the compound III with a phenol solution to obtain the novel phenolic compound;
preferably, the molar ratio of the compound III to the phenol is 1-3: 5-7, in order to mix the substrate and the phenol more uniformly.
(1) When R ═ Br, the compound III is dissolved in a phenol solution at 25-65 ℃ and N2Adding toluene sulfonic acid under the protection effect and stirring for reaction.
The phenol is solid at low temperature, and the phenol is added, so that when the temperature is increased to 25-65 ℃ (more preferably 60-65 ℃), the compound III and the phenol are mixed uniformly and are easy to react; the product has more phenolic hydroxyl groups, and nitrogen protection is added to prevent the product from being oxidized during heating; p-toluenesulfonic acid is a commonly used catalyst for such reactions (analogous to the synthesis of bisphenol a), among others sulfuric acid, hydrochloric acid, trifluoromethanesulfonic acid, etc.; we screened these acids in the early days and found that TsOH, toluene sulfonic acid, reacted best for this type of substrate.
(2) When R is Cl, the compound III is dissolved in a phenol solution, and toluenesulfonic acid is added at 25-65 ℃ and the reaction is stirred. In this case, N is not required to be added2And (4) protecting.
Wherein the chemical general formula of the compound II is shown as the following formula (2); the chemical general formula of the compound III is shown as a formula (3);
in the formulae (2) and (3), R1 ═ Br or Cl.
2. When R ═ F in the novel phenolic compounds of formula (1), the preparation process comprises:
step 1, reacting 4-Br-2, 6-difluorophenol with bromotoluene to obtain 5-Br-1, 3-difluoro-2-phenyl-methoxybenzene; specifically, 4-Br-2, 6-difluorophenol was dissolved in DMF solution in N2Then adding alkaline substance, stirring, and adding bromotoluene.
The nitrogen protection is added to prevent the product from being oxidized when being heated; the alkaline substance comprises NaOH and K2CO3Etc., K is selected for this embodiment2CO3Is in consideration of K2CO3Can completely meet the reaction requirement, is cheap and easy to obtain, and is convenient for post-treatment.
Preferably, the molar ratio of the 4-Br-2, 6-difluorophenol to the bromotoluene is 40-50: 50-60. Bromotoluene has strong irritation, can promote lacrimation, and is not beneficial in addition; this range of ratios is sufficient to complete the reaction.
Step 2, reacting the 5-Br-1, 3-difluoro-2-phenyl-methoxybenzene with benzyl magnesium chloride to obtain 4-benzyl-2, 6-dichloro-methoxybenzene;
specifically, to mix ZnBr2In THF of (1) at N2Adding the benzyl magnesium chloride at the temperature of 10 ℃, stirring and mixing uniformly, and adding a THF solution of 5-Br-1, 3-difluoro-2-phenyl-methoxybenzene. The reason why the benzylmagnesium chloride is added under such conditions is that the benzylmagnesium chloride can be rapidly converted into the organic Zn reagent, and the generated Zn reagent can be stably present and cannot be deteriorated due to too strong price adjustment.
Preferably, the molar ratio of benzylmagnesium chloride to compound 7 is in the range of 1: 1-2. This range of ratios is sufficient to complete the reaction.
Step 3, carrying out reduction reaction on the 4-benzyl-2, 6-dichloro-methoxybenzene to obtain 4-benzyl-2, 6-difluorophenol; specifically, 4-benzyl-2, 6-dichloro-methoxybenzene is dissolved in methanol solution, and H is introduced under the condition of catalyst Pd/C2And carrying out reduction reaction.
Step 4, reacting the 4-benzyl-2, 6-difluorophenol with acetyl chloride to obtain 4- (4-acetyl benzyl) -2, 6-difluorophenylacetic acid; specifically, the 4-benzyl-2, 6-difluorophenol is dissolved in dichloromethane, acetyl chloride is added dropwise, and a catalyst AlCl is added simultaneously3. The solvent for the reaction is most commonly dichloromethane, dichloroethane, carbon disulfide and the like; the catalyst for the reaction is most commonly aluminum trichloride, anhydrous zinc chloride, ferric trichloride, titanium tetrachloride and the like; the reaction selects dichloromethane as solvent, AlCl3The catalyst is obtained at low cost and easily available, and the yield is also suitable for scale-up.
Preferably, the molar ratio of the 4-benzyl-2, 6-difluorophenol to the acetyl chloride is preferably in the range of 1-2: 1.1-4, can make the reaction complete.
And 5, reacting the 4- (4-acetyl benzyl) -2, 6-difluorophenylacetic acid with a phenol solution to obtain the novel phenolic compound. P-toluenesulfonic acid is a commonly used catalyst for such reactions (analogous to the synthesis of bisphenol a), among others sulfuric acid, hydrochloric acid, trifluoromethanesulfonic acid, etc., which were previously screened by the applicant and TsOH was found to react best with such substrates. Preferably, the molar ratio of the 4- (4-acetyl benzyl) -2, 6-difluorophenylacetic acid to the phenol is 1-3: 5-7, in order to mix the substrate and the phenol more uniformly and completely react.
The novel phenolic compound can obviously inhibit the growth of gram-positive bacteria, and the minimum inhibitory concentration of the novel phenolic compound to staphylococcus aureus, bacillus subtilis and methicillin-resistant staphylococcus aureus is 1-4 mu g/mL
The following will describe a novel phenolic compound of the present application, its preparation method and application in detail with reference to examples and experimental data.
Example 1
When R ═ Br in formula (1), the novel phenolic compounds in this example have the formula:
the specific preparation method comprises the following steps:
step 1, add Compound 1 (4-hydroxydiphenylmethane, 3.00g,16.3mmol,1.00eq) to AcOH (15.0mL), and drop-add Br at 10 deg.C2(5.23g,32.7mmol,1.69mL,2.01 eq). The mixture was stirred at 5-25 ℃ for 2 hours.
After completion of the reaction of 4-hydroxydiphenylmethane by TLC (petroleum ether: ethyl acetate: 5: 1, Rf: 0.59), the reaction mixture was poured into two volumes of ice water and extracted twice with the same volume of ethyl acetate, and the resulting organic phase was extracted with NaHCO3Washing twice, then washing with strong brine and anhydrous Na2SO4Drying, filtering under reduced pressure, concentrating to obtain residue, purifying by column chromatography to obtain yellowish oily solution A, to obtain compound II, and detecting the compound by nuclear magnetic resonance;
step 2, Compound 2(3.00g,8.77mmol,1.00eq) was added dropwise to dichloromethane DCM (30.0mL), acetyl chloride (1.45g,18.4mmol,1.31mL,2.10eq), 0-15 deg.CStirring for 1 hour to obtain a mixture, and adding AlCl3(1.87g,14.0mmol,767uL,1.60eq) was added proportionally to the mixture and stirred at 0-15 ℃ for 12 hours.
After detecting that compound II is completely reacted by TLC (ethyl acetate: 5: 1, Rf: 0.30), pouring the reaction mixture into ice water, extracting with dichloromethane, washing the obtained extracted organic phase with concentrated brine, drying anhydrous Na2SO4, filtering and concentrating under reduced pressure to obtain a residue, and purifying by silica gel column chromatography to obtain a white solid B, i.e. compound 3;
step 3, TsOH (2.02g,11.7mmol,2.00eq) was added to a mixture of compound 3(2.50g,5.87mmol,1.00eq), phenol (2.76g,29.3mmol,2.58mL,5.00eq) was added, and the mixture was stirred in N2The temperature is 5-25 ℃. The mixture was stirred at 65 ℃ for 12 hours. HPLC (ET28324-9-P1a1) and TLC (petroleum ether: ethyl acetate ═ 5: 1, Rf ═ 0.51) showed complete consumption of compound 3. The reaction mixture was dissolved in ethyl acetate EtOAc (15.0mL) with the same volume of NaHCO3Washing with the solution, then washing the organic phase with half volume of concentrated brine, anhydrous Na2SO4Drying, filtering under reduced pressure, concentrating to obtain residue, purifying by column chromatography to obtain yellow solid, and further purifying by reverse high performance liquid chromatography to obtain light yellow solid to obtain the novel phenolic compound.
Example 2
When R ═ Cl in formula (1), the novel phenolic compounds of this example have the formula:
the specific preparation method comprises the following steps:
step 1, add compound 1(3.00g,16.3mmol,1eq) in MeOH (50.0mL), add NaOH (977mg,24.4mmol,1.5eq) and NaCl (2.85g,48.9mmol,3eq) at 5-15 ℃ to give a mixture. NaClO (45.5g,48.9mmol,37.6mL, 8% purity, 3eq) was added dropwise to the mixture and stirred at 5-15 ℃ for 13 h.
TLC (petroleum ether: ethyl acetate ═ 5: 1) indicated that reactant 1 had been completely consumed. The mixture was concentrated under reduced pressure to remove most of the MeOH. Diluted with water (15.0mL) and extracted with EtOAc (20.0mL × 2). The combined organic layers were washed with brine (15.0mL) over anhydrous Na2SO4Dried, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether/ethyl acetate 100/1-10/1). Compound 4(2.50g,9.88mmol, 60.7% yield) was identified by HNMR (ET25545-13-P1A) as a pale yellow solid.
Step 2, add compound 4(500mg,1.98mmol,1eq) to DCM (3mL), add acetyl chloride (325mg,4.15mmol,296uL,2.1eq) dropwise at 5 ℃, stir at 0-15 ℃ for 1 hour to give a mixture, add AlCl3(579mg,4.35mmol,237uL,2.2eq) was added to the mixture in proportion and stirred at 0-15 ℃ for 2 hours.
TLC (petroleum ether: ethyl acetate 5: 1) indicated complete consumption of reaction 4, the reaction mixture was poured into ice water (5ml) at 10 ℃ and extracted with dichloromethane DCM (2ml × 2). The combined organic layers were washed with brine (2ml) over anhydrous Na2SO4Dried, filtered and concentrated under reduced pressure to give a residue. The residue was purified by silica gel column chromatography to give Compound 5(400g,1.19mmol, 60.1% yield) which was identified as a pale yellow solid by HNMR (ET 25545-16-P1B).
Step 3, compound 5(400mg,1.19mmol,1eq) as a solution in phenol (558mg,5.93mmol,522uL,5eq) was added to TsOH (409mg,2.37mmol,2 eq). The mixture was stirred at 60 ℃ for 12 hours. LCMS (ET25545-19-P1A1) and HPLC (ET25545-19-P1A) showed complete consumption of Compound 5 and a major peak was detected. The reaction mixture was dissolved in ethyl acetate EtOAc (5.00mL) with the same volume of NaHCO3Washing with the solution, then washing the organic phase with half volume of concentrated brine, anhydrous Na2SO4Drying, filtering and concentrating under reduced pressure to obtain residue. High performance liquid chromatography (column: Welch xc 18250 mm 10 um; mobile phase: [ water ](10mm-NH4HCO3)-ACN](ii) a B%: 55% -75% for 10min) to obtain a white solid, and then obtaining the novel phenolic compound.
Example 3
When R ═ F in formula (1), the novel phenolic compounds of this example have the formula:
the specific preparation method comprises the following steps:
step 1, to Compound 6(10g,47.85mmol,1eq) in DMF (100mL) in N2Adding K at 15 deg.C2CO3(9.92g,71.77mmol,1.5eq), after stirring at 15 ℃ for 0.5 h, bromotoluene (9.82g,57.42mmol,6.82mL,1.2eq) was added and stirring at 15 ℃ for 12 h gave a mixture.
TLC (petroleum ether: ethyl acetate 3: 1) indicated complete consumption of reaction 6, the mixture was poured into 300 ml of ice water and extracted with EtOAc (100ml × 2). The combined organic layers were washed with brine (50mL x 3) and washed with anhydrous Na2SO4Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO)2Petroleum ether/ethyl acetate 100/1-20/1) to purify the residue. Compound 7(10g,33.43mmol, yield 69.87%) was a pale yellow oil as determined by HNMR (ET 25545-5-P1A).
Step 2, mixing ZnBr2(902.60mg,4.01mmol,200.58uL,2.4eq) in THF (10mL) in N2Benzylmagnesium chloride (1M,3.34mL,2.0eq) was added dropwise at 10 ℃ and stirred at 15 ℃ for 0.5 h to give a mixture. Compound 7(0.5g,1.67mmol,1eq) and 1, 3-bis (2, 6-diisopropylphenyl) -2 h-imidazole, 3-chloropyridine, dichloropalladium (113.81mg,167.00umol,0.1eq) were dissolved in THF (5mL) in N2Added dropwise to the mixture at 15 ℃ and stirred at 15 ℃ for 12.5 hours.
LCMS (ET25545-10-P1A) shows combinationObject 7 is completely consumed and an ideal m/z peak is detected. The mixture was poured into ice water (20ml) and extracted with EtOAc (10ml x 2). The combined organic extracts were washed with brine (10ml), dried over MgSO4, filtered and concentrated in vacuo to give a residue. By column chromatography (SiO)2Petroleum ether/ethyl acetate 50/1-5/1) to purify the residue. Compound 8(0.3g, crude) was a yellow oil.
Step 3, Add Compound 8(0.3g,966.69umol,1eq) to MeOH (3mL), add Pd/C (0.3g, 10% purity) at 15 deg.C, in H2(15Psi), stirring at 15 ℃ for 12 hours. LCMS (ET25545-33-P1A) indicated consumption of Compound 8 and an ideal MS peak was detected. The reaction mixture was filtered and concentrated under lower pressure to give a residue, and compound 9(0.2g, crude oil) was obtained as a tan oil.
Step 4, add compound 9(0.2g,908.21umol,1eq) to DCM (3ml), add acetyl chloride (149.71mg,1.91mmol,136.10uL,2.1eq) dropwise at 5 deg.C, stir at 5 deg.C for 1 hour, add AlCl3(266.42mg,2.00mmol,109.19uL,2.2eq) and stirred at 15 ℃ for 2h to give a reaction mixture. TLC (petroleum ether: ethyl acetate ═ 5: 1) and LCMS (ET25545-40-P1A) indicated complete consumption of compound 9. The reaction mixture was poured into ice water (5ml) at 10 ℃ and then extracted with DCM (2ml × 2). The combined organic layers were washed with brine (2ml) over anhydrous Na2SO4Dried, filtered and concentrated under reduced pressure to give a residue. With pretlc (SiO)2Petroleum ether/ethyl acetate 5/1) to give compound 10(0.3g, crude) as a pale yellow solid.
Step 5, TsOH (226.36mg,1.31mmol,2.0eq) was added to a solution of compound 10(0.2g,657.27umol,1eq) dissolved in phenol (309.28mg,3.29mmol,289.05uL,5.0eq) and the mixture was stirred at 60 ℃ for 12 hours to give a reaction mixture. LCMS (ET25545-46-P1B) and HPLC (ET25545-46-P1A) showed complete consumption of Compound 10 and detected one major peak. The reaction mixture was dissolved in EtOAc (5mL) with the same volume of NaHCO3Washing with the solution, then washing the organic phase with half volume of concentrated brine, anhydrous Na2SO4Drying, filtering and concentrating under reduced pressure to obtain residue. Subjecting the crude product to reversed phase high performance liquid chromatography (0.1% NH)4HCO3) Purification gave a pale yellow solid, which was detected by LCMS (ET25545-46-P1B1) and HPLC (ET 25545-46-P1B). The resulting solid was subjected to prep-HPLC (column: Waters Xbridge BEH C18100 30mM 10 um; mobile phase: [ water (10mM NH. sub.H.)4HCO3)-ACN](ii) a B%: 30% -80%, 10min) was given as a white solid. HNMR determination shows that the compound (0.12g,274.68umol, yield 41.79% and purity 98.99%) is white solid, i.e. the novel phenolic compound.
Comparative example 1
The comparative example is an unsubstituted phenolic compound prepared by a conventional method and having the following structural formula:
test examples
The novel phenolic compounds of examples 1 to 3 and the unsubstituted phenolic compound of comparative example 1 were subjected to Minimum Inhibition Concentration (MIC) test, and the MIC measurement results are shown in table 1.
The specific experimental steps are as follows:
inoculating a little strain from the plate into a conical flask filled with LB culture medium (Luria-Bertani culture medium), and culturing for 8-10h at 37 ℃ by shaking; wherein, the fungus comprises: gram-positive and gram-negative bacteria; the gram-positive bacteria comprise staphylococcus aureus, bacillus and methicillin-resistant staphylococcus aureus, and the bacillus subtilis is bacillus subtilis 168; the Staphylococcus aureus is Staphylococcus aureus 25923; the methicillin-resistant staphylococcus aureus is abbreviated as MRSA; the gram-negative bacterium used was Escherichia coli DH5 α.
The formulation of LB medium was as follows: 10g/L of Tryptone (Tryptophan), 5g/L of Yeast extract (Yeast extract) and 10g/L of sodium oxide (NaCl).
The inoculum was aspirated from the flask in the morning on day 2 at a volume ratio of 1% and transferred to a flask containing 20mL LB medium, and the flask was further incubated on a shaker at 37 ℃ for 5h to 6h until the OD600(OD600 represents the absorbance of the inoculum at 600 nm) was 0.6 to 0.8, and the specific OD (optical density) of the inoculum was recorded.
The cultured bacterial liquid was diluted 1000-fold with LB medium to a bacterial liquid concentration of 105CFU/ml, and the diluted bacterial liquid was aspirated and added to a 96-well plate at 150. mu.l per well.
Prepare the phenol compound solution at a concentration of 128. mu.g/ml, pipette 150. mu.l of the triphenol compound solution into well 1 of a 96-well plate, then pipette 150. mu.l into well 2, and so on. After the addition, the 96-well plate was placed in a shaker and incubated overnight at 37 ℃.
TABLE 1 MIC value results
As can be seen from table 1, the novel phenolic compounds of the present invention have a 1-fold higher inhibitory effect against gram-positive bacteria than comparative example 1. The novel phenolic compounds provided by the embodiments 1-3 of the invention can obviously inhibit the growth of gram-positive bacteria, the minimum inhibition concentration of the novel phenolic compounds to staphylococcus aureus, bacillus subtilis and methicillin-resistant staphylococcus aureus is 1-2 mu g/ml, and the trisphenol compounds have good antibacterial effect and can be used as good antibacterial agents of gram-positive bacteria.
Finally, it should also be noted that the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus.
While preferred embodiments of the present invention have been described, additional variations and modifications in those embodiments may occur to those skilled in the art once they learn of the basic inventive concepts. Therefore, it is intended that the appended claims be interpreted as including preferred embodiments and all such alterations and modifications as fall within the scope of the invention. It will be apparent to those skilled in the art that various changes and modifications may be made in the present invention without departing from the spirit and scope of the invention. Thus, if such 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 such modifications and variations.
Claims (10)
1. A novel phenolic compound having the chemical formula:
wherein, R ═ Br, Cl or F.
2. A process for producing a novel phenolic compound according to claim 1, wherein when R ═ Br or Cl is contained in the novel phenolic compound of formula (1), the process comprises:
reacting 4-hydroxydiphenylmethane with a halogen-containing compound to obtain a compound II, wherein the halogen comprises bromine or chlorine;
reacting the compound II with acetyl chloride to obtain a compound III;
reacting the compound III with a phenol solution to obtain the novel phenolic compound;
wherein the chemical general formula of the compound II is a formula (2); the chemical general formula of the compound III is shown as a formula (3);
in the formulae (2) and (3), R1 ═ Br or Cl.
3. The method of claim 2, wherein reacting 4-hydroxydiphenylmethane with a halogen-containing compound comprises:
dissolving 4-hydroxy diphenylmethane and a halogen-containing compound in an acetic acid or methanol solvent, and reacting at the temperature of 5-25 ℃.
4. The method of claim 2, wherein reacting the compound II with acetyl chloride comprises:
dissolving the compound II and acetyl chloride in a dichloromethane solvent, and reacting in a catalyst AlCl3And reacting at the temperature of 5-25 ℃.
5. The method of claim 2, wherein reacting the compound III with a phenol solution comprises:
and dissolving the compound III in a phenol solution, adding toluenesulfonic acid at the temperature of 25-65 ℃, and stirring for reaction.
6. A process for producing the novel phenolic compound of claim 1, wherein when R ═ F in the novel phenolic compound of formula (1), the process comprises:
reacting 4-Br-2, 6-difluorophenol with bromotoluene to obtain 5-Br-1, 3-difluoro-2-phenyl-methoxybenzene;
reacting the 5-Br-1, 3-difluoro-2-phenyl-methoxybenzene with benzyl magnesium chloride to obtain 4-benzyl-2, 6-dichloro-methoxybenzene;
carrying out reduction reaction on the 4-benzyl-2, 6-dichloro-methoxybenzene to obtain 4-benzyl-2, 6-difluorophenol;
reacting the 4-benzyl-2, 6-difluorophenol with acetyl chloride to obtain 4- (4-acetyl benzyl) -2, 6-difluorophenylacetic acid;
and (3) reacting the 4- (4-acetyl benzyl) -2, 6-difluorophenylacetic acid with a phenol solution to obtain the novel phenolic compound.
7. The method of claim 6, wherein reacting 4-Br-2, 6-difluorophenol with bromotoluene comprises:
4-Br-2, 6-difluorophenol was dissolved in DMF solution and washed with N2Adding K under atmosphere2CO3Stirring, and adding bromotoluene for reaction.
8. The method of claim 6, wherein reacting the 5-Br-1, 3-difluoro-2-phenyl-methoxybenzene with benzylmagnesium chloride comprises:
reacting ZnBr2Mixing with THF to obtain a first mixed solution;
in N2And mixing and stirring the benzyl magnesium chloride and the first mixed solution under the atmosphere, and adding 5-Br-1, 3-difluoro-2-phenyl-methoxybenzene to react after stirring.
9. Use of the novel phenolic compound of claim 1 for the preparation of gram-positive antibacterial agents.
10. The use of claim 9, wherein the gram positive bacteria comprise staphylococcus aureus, bacillus, methicillin-resistant staphylococcus aureus.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010298193.9A CN111574334B (en) | 2020-04-16 | 2020-04-16 | Novel phenolic compound and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010298193.9A CN111574334B (en) | 2020-04-16 | 2020-04-16 | Novel phenolic compound and preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN111574334A true CN111574334A (en) | 2020-08-25 |
| CN111574334B CN111574334B (en) | 2023-02-10 |
Family
ID=72089884
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010298193.9A Active CN111574334B (en) | 2020-04-16 | 2020-04-16 | Novel phenolic compound and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111574334B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113004144A (en) * | 2021-03-09 | 2021-06-22 | 西安瑞联新材料股份有限公司 | Synthesis method of perfluorobisphenol A |
| CN115215734A (en) * | 2022-08-17 | 2022-10-21 | 辽宁靖帆新材料有限公司 | Preparation method of 1- [4- (4-hydroxyphenyl) phenyl ] -1, 1-bis (4-hydroxyphenyl) ethane |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20030229079A1 (en) * | 1999-06-25 | 2003-12-11 | Payne Linda S. | 1-(Aromatic- or heteroaromatic-substituted)-3-(heteroaromatic substituted)-1,3-propanediones and uses thereof |
| WO2010011584A2 (en) * | 2008-07-25 | 2010-01-28 | Merck & Co., Inc. | Process for making a renin inhibitor |
| CN102596873A (en) * | 2009-09-09 | 2012-07-18 | 本州化学工业株式会社 | Novel trisphenol compound |
| CN102942454A (en) * | 2012-11-29 | 2013-02-27 | 太仓市茜泾化工有限公司 | Preparation method of 1,1,1-tri(4-hydroxyphenyl)ethane |
| CN107912434A (en) * | 2017-12-08 | 2018-04-17 | 江汉大学 | The application of three phenolic compounds and anti-biotic material |
| CN108640845A (en) * | 2018-03-29 | 2018-10-12 | 大连九信精细化工有限公司 | A method of preparing 2- [4- (2- ethoxyethyl groups) phenoxy group] ethylamine |
| CN110092722A (en) * | 2019-06-03 | 2019-08-06 | 常州市天华制药有限公司 | A method of catalyzing and synthesizing 4- acyl group aralkylphenol derivative |
| CN110475799A (en) * | 2017-04-14 | 2019-11-19 | 沙特基础工业全球技术有限公司 | The synthesis of phosphorous-containing monomers and polycarbonate is incorporated by interfacial polymerization |
-
2020
- 2020-04-16 CN CN202010298193.9A patent/CN111574334B/en active Active
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20030229079A1 (en) * | 1999-06-25 | 2003-12-11 | Payne Linda S. | 1-(Aromatic- or heteroaromatic-substituted)-3-(heteroaromatic substituted)-1,3-propanediones and uses thereof |
| WO2010011584A2 (en) * | 2008-07-25 | 2010-01-28 | Merck & Co., Inc. | Process for making a renin inhibitor |
| CN102596873A (en) * | 2009-09-09 | 2012-07-18 | 本州化学工业株式会社 | Novel trisphenol compound |
| CN102942454A (en) * | 2012-11-29 | 2013-02-27 | 太仓市茜泾化工有限公司 | Preparation method of 1,1,1-tri(4-hydroxyphenyl)ethane |
| CN110475799A (en) * | 2017-04-14 | 2019-11-19 | 沙特基础工业全球技术有限公司 | The synthesis of phosphorous-containing monomers and polycarbonate is incorporated by interfacial polymerization |
| CN107912434A (en) * | 2017-12-08 | 2018-04-17 | 江汉大学 | The application of three phenolic compounds and anti-biotic material |
| CN108640845A (en) * | 2018-03-29 | 2018-10-12 | 大连九信精细化工有限公司 | A method of preparing 2- [4- (2- ethoxyethyl groups) phenoxy group] ethylamine |
| CN110092722A (en) * | 2019-06-03 | 2019-08-06 | 常州市天华制药有限公司 | A method of catalyzing and synthesizing 4- acyl group aralkylphenol derivative |
Non-Patent Citations (2)
| Title |
|---|
| LASSALAS, PIERRIK ET AL: "Structure Property Relationships of Carboxylic Acid Isosteres", 《JOURNAL OF MEDICINAL CHEMISTRY》 * |
| 张文福主编: "《医学消毒学》", 31 July 2002, 军事医学科学出版社 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113004144A (en) * | 2021-03-09 | 2021-06-22 | 西安瑞联新材料股份有限公司 | Synthesis method of perfluorobisphenol A |
| CN113004144B (en) * | 2021-03-09 | 2022-09-30 | 西安瑞联新材料股份有限公司 | Synthesis method of perfluorobisphenol A |
| CN115215734A (en) * | 2022-08-17 | 2022-10-21 | 辽宁靖帆新材料有限公司 | Preparation method of 1- [4- (4-hydroxyphenyl) phenyl ] -1, 1-bis (4-hydroxyphenyl) ethane |
| CN115215734B (en) * | 2022-08-17 | 2024-01-05 | 辽宁靖帆新材料有限公司 | Preparation method of 1- [4- (4-hydroxyphenyl) phenyl ] -1, 1-bis (4-hydroxyphenyl) ethane |
Also Published As
| Publication number | Publication date |
|---|---|
| CN111574334B (en) | 2023-02-10 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111574334B (en) | Novel phenolic compound and preparation method and application thereof | |
| CN1819986B (en) | Preparation of 1,3-diphenylprop-2-en-1-one derivatives | |
| CN114380675B (en) | Method for synthesizing aryl phenol by reacting halogenated aromatic hydrocarbon and phenol compound under induction of visible light | |
| CN104558076A (en) | Preparation method of tildipirosin and intermediate compounds of tildipirosin | |
| Nagawade et al. | Zirconium (IV) chloride-catalysed reaction of indoles: An expeditious synthesis of Bis (indolyl) methanes | |
| CN1258274A (en) | Method for preparation of carbamates | |
| CN114524795A (en) | Improved preparation method of Rhodomyrtone | |
| CN115260050B (en) | Method for preparing 3-bromo-N-aryl propionamide by using NBS | |
| CN1185192C (en) | Preparation method of oxoisophorone | |
| CN108707067B (en) | Preparation method of 1-aryl-4, 4, 4-trifluoro-1-butanone compound | |
| CN113651829A (en) | Spiro-2, 3-dihydroquinoline-4-ketone-3, 4-dihydrocoumarin bioactive skeleton and synthesis method and application thereof | |
| CN113024517A (en) | Method for preparing erdastinib | |
| CN114560832B (en) | Method for synthesizing dibenzofuran compound | |
| AU2005257483B2 (en) | Method for producing (Z)-1-phenyl-1-diethylaminocarbonyl-2-hydroxymethyl cyclopropane | |
| Nagasawa et al. | A novel and efficient synthesis of 5-substituted and 5, 6-disubstituted 4-methylcoumarins using organocerium (III) reagents | |
| JP4879030B2 (en) | Method for producing naphthopyran compound | |
| CN108892604A (en) | A method of preparing halogenated benzo [b] Fluorenone series compound | |
| CN114736183A (en) | Preparation method of 3-methylflavone-8-carboxylic acid | |
| CN110483361B (en) | beta-C (sp)3) Synthesis of (E) -H functionalized and aromatized pyrroles | |
| CN118359569A (en) | Linderane sesquiterpene intermediate, linderane sesquiterpene polymer prepared from intermediate and preparation method | |
| Kubo et al. | Synthesis of cyclic poly (methyl methacrylate) by the intramolecular cyclization of α-amino, ω-carboxyl heterodifunctional poly (methyl methacrylate) | |
| Villemin et al. | Optimisation of solvent free parallel synthesis under microwave irradiation: synthesis of new arylacrylonitriles | |
| CN112047885B (en) | Gamma- (9-acridine) diazo acetoacetate, gamma- (9-acridine methylene) -beta-keto ester and preparation method thereof | |
| CN112358460A (en) | Long-afterglow light-storing organic luminescent material and preparation method thereof | |
| CN1185240C (en) | Process for one-stage synthesizing substituted diaryl organic metal phosphate |
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 |