CN112007074A - Oil-in-water type cinnamomum camphora oil nanoemulsion preparation and preparation method thereof - Google Patents

Oil-in-water type cinnamomum camphora oil nanoemulsion preparation and preparation method thereof Download PDF

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CN112007074A
CN112007074A CN201910458643.3A CN201910458643A CN112007074A CN 112007074 A CN112007074 A CN 112007074A CN 201910458643 A CN201910458643 A CN 201910458643A CN 112007074 A CN112007074 A CN 112007074A
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oil
percent
preparation
cinnamomum camphora
surfactant
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欧阳五庆
郑寅
魏云鹏
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Northwest A&F University
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/54—Lauraceae (Laurel family), e.g. cinnamon or sassafras
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
    • A61K47/10—Alcohols; Phenols; Salts thereof, e.g. glycerol; Polyethylene glycols [PEG]; Poloxamers; PEG/POE alkyl ethers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/26—Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharides; Derivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/44—Oils, fats or waxes according to two or more groups of A61K47/02-A61K47/42; Natural or modified natural oils, fats or waxes, e.g. castor oil, polyethoxylated castor oil, montan wax, lignite, shellac, rosin, beeswax or lanolin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0014—Skin, i.e. galenical aspects of topical compositions
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/10—Dispersions; Emulsions
    • A61K9/107—Emulsions ; Emulsion preconcentrates; Micelles
    • A61K9/1075—Microemulsions or submicron emulsions; Preconcentrates or solids thereof; Micelles, e.g. made of phospholipids or block copolymers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]

Abstract

The invention discloses an oil-in-water type camphor oil nanoemulsion preparation, which is prepared from 0.10-20.00% of camphor oil, 20.00-45.00% of surfactant, 2.00-20.00% of cosurfactant, 0-20.00% of oil phase and the balance of distilled water, wherein the sum of the mass percentages of the components is 100%. The camphor oil nanoemulsion preparation is colorless or yellow transparent in appearance, has the particle size of 1-100 nm, and is good in stability, high in safety, low in viscosity, simple in preparation method, stable in storage and good in transdermal property. Can be directly smeared on the surface of skin clinically, and is extremely convenient to use.

Description

Oil-in-water type cinnamomum camphora oil nanoemulsion preparation and preparation method thereof
Technical Field
The invention belongs to the field of medicines, relates to a new formulation of camphor oil, and particularly relates to an oil-in-water type camphor oil nanoemulsion preparation and a preparation method thereof.
Background
The linalool oil is colorless or yellowish to reddish brown oily liquid, is dissolved in ethanol and ether, has strong camphor smell, is a high ash substance and has strong volatility. The linaloe oil has antiinflammatory, analgesic, anti-depression, antibacterial, parasite killing, laxative, skin warming, exciting, sweat promoting, insect expelling, wound healing, mosquito and insect repelling, antibacterial, antipruritic, foot odor eliminating, tinea pedis preventing, and air purifying effects. In practical application, the cinnamomum camphora oil is usually used for resisting inflammation, easing pain, preventing mosquitoes and expelling parasites. Most of the camphor products used at present are chemical synthetic products, and the dosage forms are also limited to solid pills, tablets and block preparations, or the camphor products are matched with other medicines to be used as liniments or ointment for application. But the camphor has strong volatility, is easy to lose in the production and storage processes of the traditional preparation, has poor stability and influences the drug effect.
Disclosure of Invention
Aiming at the defects and shortcomings in the prior art, the invention aims to provide an oil-in-water type cinnamomum camphora oil nanoemulsion preparation with the effects of resisting bacteria, diminishing inflammation, repelling mosquitoes, preventing insects and refreshing brain. According to the invention, the cinnamomum camphora oil is prepared into the nanoemulsion, so that the transdermal absorption capacity of the cinnamomum camphora oil is enhanced, and the problems of volatility and poor stability of the cinnamomum camphora oil in the actual production, transportation and storage processes are solved.
The technical scheme for realizing the aim of the invention is that the oil-in-water type cinnamomum camphora oil nanoemulsion preparation has the particle size of 1-100 nm and consists of the following raw materials:
0.10 to 20.00 percent of linaloe oil, 20.00 to 45.00 percent of surfactant, 2.00 to 20.00 percent of cosurfactant, 0 to 20.00 percent of oil phase and the balance of distilled water, wherein the sum of the mass percent of the components is 100 percent.
The oil-in-water type cinnamomum camphora oil nanoemulsion preparation disclosed by the invention has the following excellent mixture ratio: 5.00 to 18.00 percent of linaloe oil, 25.00 to 40.00 percent of surfactant, 10.00 to 20.00 percent of cosurfactant, 2.00 to 10.00 percent of oil phase and the balance of distilled water, wherein the sum of the mass percentages of the components is 100 percent.
The oil-in-water type cinnamomum camphora oil nanoemulsion preparation of the invention has the following further optimal mixture ratio: 10.00 to 15.00 percent of linaloe oil, 30.00 to 40.00 percent of surfactant, 15.00 to 20.00 percent of cosurfactant, 5.00 to 8.00 percent of oil phase and the balance of distilled water, wherein the sum of the mass percentages of the components is 100 percent.
The surfactant is one or a mixture of more of castor oil polyoxyethylene ether (EL-40), hydrogenated castor oil polyoxyethylene ether (RH-40), tween-80 (tween-80) and span-80 (span-80).
The cosurfactant is any one of ethanol, 1, 2-propylene glycol, glycerol and 1, 3-butanediol.
The oil phase is one or more of isopropyl myristate (IPM), wheat germ oil, liquid paraffin, ethyl acetate, vitamin E oil, and edible soybean oil.
The used linalool oil can be used as an oil phase, and can also form a nano-emulsion with a surfactant, a cosurfactant and water in a formula without adding other oil phases.
Another object of the present invention is to provide a method for preparing the oil-in-water type linaloe oil nanoemulsion preparation, which comprises the following steps:
(1) taking linaloe oil, a surfactant, a cosurfactant, an oil phase and distilled water for later use;
(2) uniformly mixing the cosurfactant, the cinnamomum camphora oil and the oil phase to fully dissolve the cinnamomum camphora oil;
(3) adding a surfactant, and uniformly stirring to obtain a clear solution;
(4) and slowly adding distilled water, and stirring to obtain clear, transparent, low-viscosity, and fluid colorless or yellow solution.
The oil-in-water type camphor oil nanoemulsion preparation can be further diluted into stable nanoemulsion with any concentration by water with any proportion and directly smeared on the surface of skin for use.
The oil-in-water type cinnamomum camphora oil nanoemulsion preparation consists of oil, water, a surfactant and a cosurfactant, and is an isotropic thermodynamically stable colloid dispersion system formed by dispersing emulsion droplets with the particle size of 1-100 nm in another liquid. The nano emulsion has both water phase and oil phase, has excellent dissolving performance, and can dissolve both non-polar hydrophobic medicine and polar hydrophilic medicine.
The invention discloses a theoretical basis of an oil-in-water type cinnamomum camphora oil nanoemulsion preparation: the nano-emulsion can solubilize the drug into a nano-emulsion system to improve the stability of the drug, and can also improve the skin permeability at the same time, thereby improving the drug utilization rate to the maximum extent. In addition, the invention belongs to oil-in-water type nano-emulsion, and the cinnamomum camphora oil is wrapped in the water phase, so that the volatilization of the cinnamomum camphora oil can be effectively prevented. The surfactant is a substance necessary for forming the microemulsion, and mainly plays a role in reducing interfacial tension to form an interfacial film and promoting the formation of the nanoemulsion. The choice of surfactant will depend on the nature and purpose of use of the nanoemulsion formed. The invention selects non-ionic surfactant with non-toxicity and good biocompatibility. The non-ionic surfactant is stable in solution, is not easily affected by strong electrolyte and inorganic salts, is not easily affected by acid and alkali, and has good compatibility with other surfactants and small hemolytic effect.
The oil-in-water type cinnamomum camphora oil nanoemulsion preparation is detected by a transmission electron microscope, the diameter of liquid drops is distributed between 1nm and 100nm, and the appearance is colorless or yellow transparent liquid.
The invention has good thermodynamic stability and storage stability, can not be layered after long-term storage, and has no layering phenomenon even if centrifugation is carried out at 10000rpm for 10 min.
The oil-in-water type cinnamomum camphora oil nanoemulsion preparation has the following main effects: has effects in relieving inflammation and pain, expelling mosquito and insect, refreshing brain, and treating skin inflammation, swelling and pain, and mosquito and insect bite. The application method of the invention is that a proper amount of the ointment is directly applied to the surface of skin which is inflamed or bitten by mosquitoes.
Compared with the prior art, the oil-in-water type cinnamomum camphora oil nanoemulsion preparation has the following advantages:
(1) the oil-in-water type cinnamomum camphora oil nanoemulsion preparation belongs to nano-scale emulsion, has better skin permeability, and can quickly enter the skin to play the drug effect.
(2) The camphor oil used in the oil-in-water type camphor oil nanoemulsion preparation is extracted from camphor fat, belongs to botanical drugs, and is green, safe and effective.
(3) The oil-in-water type cinnamomum camphora oil nanoemulsion preparation is colorless or yellow transparent liquid, has good light transmission, is easy to find any nonuniformity or precipitate, and improves the sensory quality;
(4) the oil-in-water type cinnamomum camphora oil nanoemulsion preparation wraps cinnamomum camphora oil in a water phase, improves the stability of the cinnamomum camphora oil, and can effectively prevent the loss of medicines in the production, storage and transportation processes caused by volatilization of the cinnamomum camphora oil.
(5) The oil-in-water type cinnamomum camphora oil nanoemulsion preparation is safe, low in viscosity, small and uniform in particle size, improves the dispersion degree of the contained cinnamomum camphora oil, and promotes the absorption of the medicine in the body;
(6) the oil-in-water type cinnamomum camphora oil nanoemulsion preparation can be directly smeared on the surface of skin, and is extremely convenient for clinical use;
(7) the preparation method of the oil-in-water type cinnamomum camphora oil nanoemulsion preparation is simple in process and suitable for large-scale industrial production.
Drawings
FIG. 1 is a transmission electron microscope image of the oil-in-water type linalool oil nanoemulsion preparation of the present invention.
Detailed Description
The beneficial effects of the oil-in-water type cinnamomum camphora oil nanoemulsion preparation are further explained by combining the curative effect test, the contrast test and the mosquito repelling effect test of the oil-in-water type cinnamomum camphora oil nanoemulsion preparation provided by the inventor.
Test example 1: curative effect test and comparative test of oil-in-water type cinnamomum camphora oil nanoemulsion preparation
First, test method
70 healthy mice were selected and randomly divided into 7 groups of 10 mice each, each half of which was male and female. The 7 groups of mice are respectively a high-dose test group, a medium-dose test group, a low-dose test group, a blank control group, a positive control group, a blank nanoemulsion (compared with the oil-in-water type camphor oil nanoemulsion preparation group, except that the camphor oil is not contained, the other components are the same) group and a common camphor oil group. The high, medium and low dosage test components respectively administer 0.3ml/cm of the oil-in-water type cinnamomum camphora oil nanoemulsion preparation of the invention2(Camphor content about 45 mg), 0.2ml/cm2(Camphor content about 30mg), 0.1ml/cm2(camphor content about 15 mg); the blank control group is given with proper amount of normal saline; yang (Yang)The sexual control group is given 1g/cm2The essential balm (Nantong Chinese medicine industry Co., Ltd., national medicine standard Z32020631, camphor content about 30 mg); the dosage of blank nanoemulsion is 0.2ml/cm2(ii) a The common linaloe oil is given by 0.2ml/cm2Linalool oil (Hubei xing Yinhe chemical Co., Ltd., camphor content is about 30 mg).
The medicines are respectively and uniformly coated on the front and back surfaces of the left ear and the right ear of a mouse, and the medicine is administered once for 1 hour and continuously for 8 hours. Xylene was used as an inflammatory agent, and after 1 hour of the last administration, the mice were anesthetized with ether, 0.02ml of the inflammatory agent was applied to the front and rear surfaces of the right ear of each mouse, and the left ear was used as a control. After 15min, the mice were sacrificed, ears were cut off, round lugs were punched at the same positions by a punch with a diameter of 9mm, and the mice were weighed, and the swelling inhibition rate was calculated by taking the weight difference between the two lugs as the swelling degree, and the inhibition rate (%) = (swelling degree of blank control group-swelling degree of experimental group)/swelling degree of blank control group × 100%. Statistical analysis was performed using SPSS11.0 analysis software.
Second, test results
The anti-inflammatory effect of the oil-in-water type cinnamomum camphora oil nanoemulsion preparation on mice is shown in table 1.
TABLE 1 anti-inflammatory action of oil-in-water type Cinnamomum camphora oil nanoemulsion formulation of the present invention on mice
Group of Average inflammatory swelling degree/mg Inhibition ratio (%)
High dose test group 1.7±0.21 89.94
Middle dose test group 1.9±0.18 88.76
Low dose test group 2.4±0.11 85.80
Blank control group 16.9±0.32 0
Positive control group 8.1±0.24 52.07
Blank nanoemulsion group 16.2±0.41 4.14
Common linaloe oil 7.5±0.15 55.62
Note: p <0.01 compared to the positive control group; compared with common linalool oil, P is less than 0.01
As can be seen from Table 1, the common camphor oil and the high, medium and low dose oil-in-water type camphor oil nanoemulsion preparation of the invention can inhibit mouse ear swelling caused by xylene, the inhibition rates are 55.62% and 89.94%, 88.76% and 85.80% respectively, and the blank nanoemulsion has no obvious anti-inflammatory effect. Compared with the common camphor oil and the positive control group, the oil-in-water type camphor oil nanoemulsion preparation has very obvious difference (P is less than 0.01), and the anti-inflammatory effect of the oil-in-water type camphor oil nanoemulsion preparation is better than that of the common camphor oil and the cooling oil.
Test example 2: mosquito repellent effect test of oil-in-water type cinnamomum camphora oil nanoemulsion preparation
First, test method
Attack force test 300 test insects were placed in a mosquito cage of 40 cm x 30 cm, with the back of the hand exposed to 4 cm x 4 cm of skin and the rest covered tightly. The hands are put into the cage for 2 min, more than 30 test insects which are stopped before come are qualified in the attacking capacity, and the test insects in the person and the cage can be used for repelling experiments.
The mosquito repellent effect test shows that 15 persons with qualified attack force are randomly divided into three groups, one group is an oil-in-water type cinnamomum camphora oil nanoemulsion preparation test group, two groups are an deet amine control group, and three groups are blank nanoemulsion control groups. The skin area of 5 cm × 5 cm is drawn on the back of each hand of the human subject, wherein one hand is a blank control and the other hand is 1.5 mg/cm2Ointment, 1.5. mu.l/cm2Uniformly coating the dose of the repellent to be tested on the liquid (aerosol is prepared into a liquid preparation) repellent, exposing the repellent for 2 min, observing whether mosquitoes suck blood or not, testing once every hour, judging that the repellent fails as long as 1 mosquito sucks blood or not, and recording the effective protection time (h) of the repellent.
Second, test results
TABLE 2 mosquito repelling effect test results
Group of Number of human subjects (human) Effective guard time (h)
The invention relates to an oil-in-water type cinnamomum camphora oil nanoemulsion preparation test group 5 6.5*#
Deet control group 5 4.0
Blank nanoemulsion control group 5 0
Note: p <0.01 compared to the diethyltoluamide control group; p <0.01 compared to blank nanoemulsion control.
The mosquito repellent effect test result shows that the effective protection time of the oil-in-water type cinnamomum camphora oil nanoemulsion test group is 6.5h, and the difference is very obvious compared with 3.9h of an diethyltoluamide control group and 0h of a blank nanoemulsion control group, so that the oil-in-water type cinnamomum camphora oil nanoemulsion preparation has a remarkable mosquito repellent effect.
Experiment 3: microscopic morphology observation of oil-in-water type cinnamomum camphora oil nanoemulsion preparation
Taking 10mL of the oil-in-water type cinnamomum camphora oil nanoemulsion preparation, diluting the preparation with distilled water by 5 times, and observing the microscopic morphology of the nanoemulsion by adopting a transmission electron microscope. The transmission electron microscope picture of the oil-in-water type cinnamomum camphora oil nanoemulsion preparation of the invention is shown in figure 1: the transmission electron microscope results show that the nano-emulsion has spherical liquid drops, uniform liquid drop size and good dispersibility.
The following are specific examples given by the inventors, but the oil-in-water type linalool oil nanoemulsion formulation of the present invention is not limited to this range.
Example 1
2.00g of absolute ethyl alcohol is uniformly mixed with 0.10g of camphor wood oil and 0.10g of wheat germ oil, so that the camphor wood oil is fully dissolved; adding RH-4020.00 g, and stirring uniformly to obtain a clear solution; and slowly adding 77.80g of distilled water, and stirring until a clear and transparent colorless or yellow solution with small viscosity and fluidity is formed.
Example 2
Linaloe oil 5.00g, liquid paraffin 1.00g, EL-4025.00 g, 1, 2-propylene glycol 5.00g, distilled water 64.00g
The specific procedure was the same as in example 1.
Example 3
10.00g of linaloe oil, 5.00g of IPM, Tween-8030.00 g, 10.00g of glycerol and 45.00g of distilled water
The specific procedure was the same as in example 1.
Example 4
15.00g of linaloe oil, 2.80g of ethyl acetate, 2.20g of liquid paraffin, 10.00g of Tween-8040.00 g of glycerol and 30.00g of distilled water
The specific procedure was the same as in example 1.
Example 5
4.00g of linaloe oil, 2.00g of edible soybean oil, RH-4035.00 g, 20.00g of glycerol and 39.00g of distilled water
The specific procedure was the same as in example 1.
Example 6
20.00g of linaloe oil, Tween-8040.00 g, 15.00g of absolute ethyl alcohol and 25.00g of distilled water
The specific procedure was the same as in example 1.
Example 7
6.00g of cinnamomum camphora oil, 4.00g of vitamin E oil, Tween-8030.00 g, Span-8010.00 g, 15.00g of 1, 3-butanediol and 35.00g of distilled water
The specific procedure was the same as in example 1.
Example 8
3.00g of cinnamomum camphora oil, 1.00g of vitamin E oil, 1.00g of Tween-8030.00 g, Span-8010.00 g, 15.00g of 1, 3-butanediol and 36.00g of distilled water
The specific procedure was the same as in example 1.
Example 9
0.10g of linaloe oil, RH-4020.00 g, 2.00g of 1, 3-butanediol and 77.90g of distilled water
The specific procedure was the same as in example 1.

Claims (4)

1. The oil-in-water type cinnamomum camphora oil nanoemulsion preparation is characterized in that the particle size of the preparation is 1 nm-100 nm, and the preparation consists of the following raw materials:
0.10 to 20.00 percent of linaloe oil
20.00 to 45.00 percent of surfactant
2.00 to 20.00 percent of cosurfactant
0 to 20.00 percent of oil phase
The balance of distilled water
The sum of the mass percentages of the components is 100 percent;
the surfactant is any one or a mixture of more of castor oil polyoxyethylene ether, hydrogenated castor oil polyoxyethylene ether, tween-80 and span-80;
the cosurfactant is any one of ethanol, 1, 2-propylene glycol, glycerol and 1, 3-butanediol;
the oil phase is any one or a mixture of more of IPM, wheat germ oil, liquid paraffin, ethyl acetate, vitamin E oil and edible soybean oil.
2. The oil-in-water linalool oil nanoemulsion formulation according to claim 1, characterized by consisting of the following raw materials:
5.00 to 18.00 percent of linaloe oil
25.00 to 40.00 percent of surfactant
Cosurfactant 10.00-20.00%
2.00 to 10.00 percent of oil phase
The balance of distilled water
The sum of the mass percentages of the components is 100 percent.
3. The oil-in-water linalool oil nanoemulsion formulation according to claim 1, characterized by consisting of the following raw materials:
linaloe oil 10.00-15.00%
30.00 to 40.00 percent of surfactant
Cosurfactant 15.00-20.00%
Oil phase 5.00-8.00%
The balance of distilled water
The sum of the mass percentages of the components is 100 percent.
4. The method of preparing the oil-in-water linalool oil nanoemulsion formulation of claim 1, comprising the following steps:
(1) weighing linalool oil, a surfactant, a cosurfactant, an oil phase and distilled water for later use;
(2) uniformly mixing the cosurfactant, the cinnamomum camphora oil and the oil phase to fully dissolve the cinnamomum camphora oil;
(3) adding a surfactant, and uniformly stirring to obtain a clear solution;
(4) and slowly adding distilled water, and stirring to obtain clear, transparent, low-viscosity, and fluid colorless or yellow solution.
CN201910458643.3A 2019-05-29 2019-05-29 Oil-in-water type cinnamomum camphora oil nanoemulsion preparation and preparation method thereof Withdrawn CN112007074A (en)

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WW01 Invention patent application withdrawn after publication
WW01 Invention patent application withdrawn after publication

Application publication date: 20201201