CN102793639A - Plant extract-mediated drug transdermal introducing system and transdermal method thereof - Google Patents
Plant extract-mediated drug transdermal introducing system and transdermal method thereof Download PDFInfo
- Publication number
- CN102793639A CN102793639A CN2012103026269A CN201210302626A CN102793639A CN 102793639 A CN102793639 A CN 102793639A CN 2012103026269 A CN2012103026269 A CN 2012103026269A CN 201210302626 A CN201210302626 A CN 201210302626A CN 102793639 A CN102793639 A CN 102793639A
- Authority
- CN
- China
- Prior art keywords
- plant extract
- transdermal
- preparation
- mediation
- nucleic acid
- 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.)
- Pending
Links
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
-
- 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
- A61K47/44—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
- 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
- A61K47/46—Ingredients of undetermined constitution or reaction products thereof, e.g. skin, bone, milk, cotton fibre, eggshell, oxgall or plant extracts
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/33—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing oxygen
- A61K8/34—Alcohols
- A61K8/342—Alcohols having more than seven atoms in an unbroken chain
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/33—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing oxygen
- A61K8/39—Derivatives containing from 2 to 10 oxyalkylene groups
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/60—Sugars; Derivatives thereof
- A61K8/606—Nucleosides; Nucleotides; Nucleic acids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9783—Angiosperms [Magnoliophyta]
- A61K8/9789—Magnoliopsida [dicotyledons]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9783—Angiosperms [Magnoliophyta]
- A61K8/9794—Liliopsida [monocotyledons]
-
- 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
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Epidemiology (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Birds (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Botany (AREA)
- Biotechnology (AREA)
- Dermatology (AREA)
- Mycology (AREA)
- Microbiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Emergency Medicine (AREA)
- General Chemical & Material Sciences (AREA)
- Molecular Biology (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Biochemistry (AREA)
- Genetics & Genomics (AREA)
- Cosmetics (AREA)
- Medicinal Preparation (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The present invention discloses a plant extract-mediated drug transdermal introducing system and a transdermal method thereof. The plant extract-mediated drug transdermal introducing system comprises a transdermal introducing agent prepared from at least one plant extract, and biological macromolecules or a preparation thereof. The transdermal method includes: using the transdermal introducing agent to promote penetration, then performing transdermal penetration for the biological macromolecules or the preparation thereof; or mixing the biological macromolecules or the preparation thereof with the transdermal introducing agent to prepare a composite formula preparation for transdermal penetration. The present invention also provides the application of the plant extract-mediated drug transdermal introducing system or the transdermal method thereof in the preparation of skin health products. Preparing skin health products by the system, the biological macromolecules can penetrate into the skin to achieve a better therapeutic effect.
Description
Technical field
The present invention relates to a kind of drug transdermal import system and transdermal methods thereof of plant extract mediation.
Background technology
Because the local skin pigment anomaly that the excessive and inhomogeneous secretion of melanin is caused is calm, can form speckle or speckle in face and the back of the hand, as: cyasma, pox seal, birthmark, nevus, freckle, sun sunburn and senile plaque etc.
Before the several centuries, people just use various external-applied ointments to come skin whitening, eliminate pigmentation.In recent years; People dispel pigmentation with chemical substances such as hydroquinone (HQ), L-ascorbic acid-2-magnesium phosphorate (MAP), hydroquinone monomethyl ether, N-acetyl group-4-S-cysteamine base phenol and arbutin; But these materials can cause side effect such as local decolouring, dermatitis and even canceration and ochronosis; Force people to have to seek succedaneum (people such as Parvez S, Phytother Res.2006,20 (11): 921-34).
Percutaneous absorption enhancer refers to promote that medicine penetrates horny layer, carries out the material of epidermis diffusion.Promoter is from chemically being divided into (Pfister W R etc., Pharm Tech, 1990,14 (9): 132) such as surfactant, lipid solvent, hydrophilic solvent and macrocyclic compound.At present, the transdermal enhancer of studying and use morely comprises natural accelerator and synthesis accelerant two types of (square generation equality, Chinese Hospitals pharmaceutical journal, 2000,20 (12): 750-752).
Natural accelerator comprises: 1) terpenoid.As pennyroyal (Mentholum (and Krishnaiah YS etc., Pharm DevTech nol, 2002,7 (3): 305), menthol, Oleum menthae), 104), ledum terpenes Borneolum Syntheticum (Zhu Jianping etc., Chinese Pharmaceutical Journal, 1990,34 (2):; 2) volatile oil or quintessence oil are like eucalyptus oil (Chen Hongqing, pharmacy progress, 2000; 235), Oleum Cinnamomi (Shen Qi etc., Chinese Hospitals medicine magazine, 2001,21 (4): 197), Fructus Cnidii volatile oil, sweetgum oil (Li Bei 24 (4):; Shanghai Medical Univ's journal, 365), Oleum Terebinthinae (Li Juan etc., China Medicine University's journal 1998,25 (5):; 343), Rhizoma Alpiniae Officinarum oil (Shen Qi etc., Chinese crude drug, 2,000 1999,30 (5):; 697), Flos Caryophylli volatile oil (Zhao K etc., J Control Release, 1998,55 (2-3): 253-260), Radix Angelicae Sinensis volatile oil etc. 23 (11):; 3) alkaloid is like Bulbus Fritillariae Thunbergii alkaloid (Ma Yunshu etc., Asia-Pacific traditional medicine, 2011 (4): 15-18); 4) lactone all has (the permanent hero of difficult ripple, foreign medical science Chinese medicine fascicle, 1993,15 (1): 41) such as dermal osmosis effect like the ligustilide in the Rhizoma Chuanxiong ether extract, senkyunolide, cnidiumm lactone, cyclobutenyl furan lactone, neocindilide.
Natural penetration enhancer all is natural plant extracts, has good transdermal enhancing effect, and zest is less, and the value of development and use is arranged very much, becomes the focus of penetration enhancer research in recent years.
(RNAinterference, RNAi) technology is the naturally occurring a kind of mechanism of removing detrimental protein of cell itself, is found by U.S. scientist Andrew Fire and Craig Mello, and obtains Nobel Prize in medicine in 2006 in the RNA interference.The mechanism of RNAi is that normal cell is on the hazard or when injuring, siRNA (small interfering RNA siRNA) can trigger the gene silencing path, cell through the RNAi path with deleterious gene " silence ".In this natural cytophylaxis process; Double-stranded RNA (dsRNA) is become the siRNA of 20-25 nucleotide by the Dicer enzyme action; (RNA-induced silencing complex RISC) combines and unwinds the inductive silencing complex of siRNA and RNA, and wherein a chain combines with target gene mRNA; Make the degraded of this gene, thereby deleterious target protein can't be synthesized and reach therapeutic purposes.
Still do not have at present and use plant extract macromolecular drugs such as nucleic acid successfully to be transported to the report of dermal layer of the skin as percutaneous introduction agent.We apply to plant extract such as Radix Angelicae Sinensis volatile oil etc. in the transdermal import system of biopharmaceutical macromolecular drugs such as nucleic acid first as percutaneous introduction agent; And successfully make the nucleic acid molecules can the percutaneous natural cover for defense; Especially horny layer, thereby through skin basal layer performance therapeutical effect.
Summary of the invention
Technical problem to be solved by this invention provides the drug transdermal import system and the transdermal methods of a kind of plant extract mediation, make biomacromolecule can the percutaneous natural cover for defense (horny layer) thus the basal layer of through skin is brought into play its therapeutical effect.
In order to achieve the above object, the invention provides a kind of drug transdermal import system of plant extract mediation, it is characterized in that, comprising: the percutaneous introduction agent that 1) prepares with at least a plant extract; 2) biomacromolecule or its preparation.
Preferably, described plant extract is a plant volatile oil.
Further, described plant volatile oil is any one or a few the mixture in Radix Angelicae Sinensis volatile oil, Oleum Folium Artemisiae Argyi, eucalyptus oil, Oleum Caryophylli, Oleum Cinnamomi, Oleum Terebinthinae and the hot thin oil.
Preferably, described plant extract is a terpenoid.
Further, described terpenoid is any one or a few the mixture in Oleum menthae, Borneolum Syntheticum and the Mentholum.
Preferably, described percutaneous introduction agent is become by the Jojoba oil of 0～98 weight portion, the Radix Angelicae Sinensis volatile oil of 1～100 weight portion, the Flos Rosae Rugosae quintessence oil of 0.2～6 weight portion, the Herba Lysimachiae foenum-graeci quintessence oil of 0～5 weight portion, the Olibanum quintessence oil of 0～10 weight portion, the tea tree ethereal oil of 0～4 weight portion and the peppermint essence line of oils of 0～0.5 weight portion.
Preferably, described biomacromolecule is nucleic acid molecules, protein molecule, peptide molecule or insulin molecule.
Further, described biomacromolecule is a nucleic acid molecules, and described biomacromolecule preparation is a nucleic acid preparation.
Further, described nucleic acid preparation comprise nucleic acid molecules and can with the preparation of its compatibility.
Further, described can be lipid or alcohols with the preparation of nucleic acid molecules compatibility.
Further, described nucleic acid molecules is functional r NA and the version that contains modification or analogies and DNA and contains the version of modification or any one or a few the mixture in the analogies.
Further, described functional r NA is any one or a few the mixture in siRNA, miRNA, little part RNA (small ligand RNA is called for short sliRNA) and the RNA aptamers.
Further, described nucleic acid preparation is the cosmetic emulsion that comprises nucleic acid molecules, comprises following component by weight percentage:
Olivem?1000 2.5～4％；
Oliwax?IC 0.5～2％；
16 octadecanol 1～6%;
Polydimethylsiloxane 2～4%;
Hydrogenation Fructus Canarii albi oleic acid Octyl Nitrite 1.5～8%;
PEG-100 stearate 0.5～4%;
Stearic acid 0.5～3%;
C15-19 alkane 3～6%;
Hydrogenated polydecene 2～7%;
Butylated hydroxytoluene 0.1～0.3%;
Butanediol 5～7%;
Glycerol 8.5～12%;
Xanthan gum 0.2～0.35%;
Hyaluronic acid 0.01～0.05%;
EDTA-disodium 0.1～0.3%;
siRNA 0.001～0.02％；
Essence 0.1～0.5%;
Antiseptic 0～1%;
Deionized water complements to 100%.
Further, described nucleic acid preparation is the cosmetics cream frost that comprises nucleic acid molecules, comprises following component by weight percentage:
Olivem?1000 3～4％；
Oliwax?IC 1～2％；
Stearic acid 1～3%;
16 octadecanol 1～6%;
PEG-100 stearate 2.5～3%;
Hydrogenated polydecene 2～7%;
Caprylic/capric triglyceride 2～6%;
Butyrospermum parkii fruit fat 1～4%;
Isopropyl palmitate 2～4%;
Hydrogenation Fructus Canarii albi oleic acid Octyl Nitrite 1～7%;
Butylated hydroxytoluene 0.2～0.25%;
Butanediol 5～7%;
Glycerol 8.5～12%;
Xanthan gum 0.25～0.35%;
Hyaluronic acid 0.01～0.05%;
EDTA-disodium 0.1～0.3%;
siRNA 0.001～0.02％；
Essence 0.1～0.5%;
Antiseptic 0～1%;
Deionized water complements to 100%.
The present invention also provides a kind of drug transdermal method of plant extract mediation; It is characterized in that; Adopt the drug transdermal import system of above-mentioned plant extract mediation, concrete steps are: 1) urge with said percutaneous introduction agent earlier, then said biomacromolecule or its preparation are carried out transdermal; Or 2) with behind said biomacromolecule or its preparation and the said percutaneous introduction agent mix homogeneously, process the compound prescription preparation, carry out transdermal.
Preferably, the compound prescription preparation said method 2) comprises following component by weight percentage:
Angelica essential oil 1～20%;
Olivem?1000 2.5～4％；
Oliwax?IC 0.5～2％；
16 octadecanol 1～6%;
Polydimethylsiloxane 2～4%;
Hydrogenation Fructus Canarii albi oleic acid Octyl Nitrite 1.5～8%;
PEG-100 stearate 0.5～4%;
Stearic acid 0.5～3%;
C15-19 alkane 3～6%;
Hydrogenated polydecene 2～7%;
Butylated hydroxytoluene 0.1～0.3%;
Butanediol 5～7%;
Glycerol 8.5～12%;
Xanthan gum 0.2～0.35%;
Hyaluronic acid 0.01～0.05%;
EDTA-disodium 0.1～0.3%;
siRNA 0.001～0.02％；
Essence 0.1～0.5%;
Antiseptic 0～1%;
Deionized water complements to 100%.
Preferably, the compound prescription preparation said method 2) comprises following component by weight percentage:
Angelica essential oil 1～20%;
Olivem?1000 3～4％；
Oliwax?IC 1～2％；
Stearic acid 1～3%;
16 octadecanol 1～6%;
PEG-100 stearate 2.5～3%;
Hydrogenation gathers the last of the ten Heavenly stems 2～7%;
Caprylic/capric triglyceride 2～6%;
Butyrospermum parkii fruit fat 1～4%;
Isopropyl palmitate 2～4%;
Hydrogenation Fructus Canarii albi oleic acid Octyl Nitrite 1～7%;
Butylated hydroxytoluene 0.2～0.25%;
Butanediol 5～7%;
Glycerol 8.5～12%;
Xanthan gum 0.25～0.35%;
Hyaluronic acid 0.01～0.05%;
EDTA-disodium 0.1～0.3%;
siRNA 0.001～0.02％；
Essence 0.1～0.5%;
Antiseptic 0～1%;
Deionized water complements to 100%.
The present invention also provides the application of drug transdermal import system in preparation skin health product of above-mentioned plant extract mediation.
Preferably, described skin health product is cosmetics.
Further, described cosmetics are the cosmetics of Pear Power spot-removing function.
Further, described cosmetics are the whitening cosmetics that contains the siRNA that regulates the melanin related gene expression.
The present invention also provides the application of drug transdermal method in preparation skin health product of above-mentioned plant extract mediation.
Preferably, described skin health product is cosmetics.
Further, described cosmetics are the cosmetics of Pear Power spot-removing function.
Further, described cosmetics are the whitening cosmetics that contains the siRNA that regulates the melanin related gene expression.
The drug transdermal import system and the transdermal methods thereof of plant extract mediation provided by the invention can be applied in the skin health product, in cosmetics, so that biomacromolecule penetrates skin, reach better therapeutic effect.
Description of drawings
Fig. 1 is a mice transdermal photo;
Fig. 2 carries out the effect contrast figure of nucleic acid transdermal for different percutaneous introduction agenies;
Fig. 3 is the angelica essential oil of variable concentrations and the effect contrast figure that Jojoba oil compound prescription preparation carries out the nucleic acid transdermal;
Fig. 4 carries out the effect contrast figure of nucleic acid transdermal for the compound prescription percutaneous introduction agent cooperates nucleic acid emulsion formulations I;
Fig. 5 carries out the effect contrast figure of nucleic acid transdermal for the compound prescription percutaneous introduction agent cooperates nucleic acid cream frost formula I;
Fig. 6 carries out the effect contrast figure of nucleic acid transdermal for the compound prescription percutaneous introduction agent cooperates nucleic acid emulsion formulations II;
Fig. 7 carries out the effect contrast figure of nucleic acid transdermal for the compound prescription percutaneous introduction agent cooperates nucleic acid cream frost formula I I;
Fig. 8 carries out the effect contrast figure of nucleic acid transdermal for different percutaneous introduction agenies cooperate nucleic acid emulsion formulations I;
Fig. 9 is the effect contrast figure of compound prescription (percutaneous introduction agent+nucleic acid molecules/nucleic acid preparation) transdermal.
The specific embodiment
For making the present invention more obviously understandable, now with preferred embodiment, and conjunction with figs. elaborates as follows.
Embodiment 1
Short the passing through of animal skin tested
1. key instrument, reagent and material
1.1ICR mice: Nantong University's Experimental Animal Center, weigh 18～20g 4～6 ages in week, male and female are at random.
1.2 the siRNA of fluorescently-labeled siRNA:Cy3 labelling (Cy3-siRNA) (Biomics Bioisystech Co., Ltd).
1.3 instrument: freezing microtome (Leica CM1900, U.S. Leica company); Just putting fluorescence microscope (X3, Japanese Olympus company) etc.
1.4 reagent: 10% (m/V) chloral hydrate (Chemical Reagent Co., Ltd., Sinopharm Group); Medical glue (the medical glue company limited of Guangzhou white clouds); Sodium chloride injection (0.9%) (the healthy and free from worry pharmaceutcal corporation, Ltd in Shandong); (0.1MPBS Biomics Bioisystech Co., Ltd); Angelica essential oil (the natural medicinal oil company limited of Jiangxi Cedrus deoclar (Roxb.) G. Don); Azone (Aladdin reagent (China) company limited).
1.5 material: 1mL disposable syringe, 5mL disposable syringe (Jiangyan City newly revive medical apparatus and instruments company limited), disposable cotton swab, 5mL centrifuge tube, eye scissors, hospital gauze etc.
2. experimental procedure
2.1 preparation percutaneous introduction agent
2.1.1 adopt the percutaneous introduction agent of angelica essential oil mediation
2.1.2 adopting PBS solution is percutaneous introduction agent, as matched group.
2.2 pretreatment:
Respectively inject 80 μ L, 10% chloral hydrate at two groups of ICR mouse peritoneals and anaesthetize, treat the mice holonarcosis after, mice is lain on the hospital gauze, use the eye scissors lagging to wipe out the about 1.5cm * 1.5cm of mouse back center hair, can not break mouse skin.At the bottom of wiping out the centrifuge tube pipe along the centrifugal mouth of pipe 1cm of 5mL place, take the centrifuge tube mouth of pipe one end, and the edge is repaired smooth.Draw the medical glue of 10uL and be coated on 5mL centrifuge tube oral area edge, then centrifuge tube is pressed on the mouse back skin, and lasting 10s, as shown in Figure 1.
Pass through 2.3 percutaneous introduction agent is short:
Dip in cotton swab and to get percutaneous introduction agent and evenly spread upon on the mouse skin in the centrifuge tube, and continue to smear 1min, repeat behind the 10min to smear once, smear altogether 3 times with cotton swab.
Smear for the third time and finish back 10min at interval, be coated with the position of percutaneous introduction agent, the remaining percutaneous introduction agent of removal skin surface with clean cotton swab wiping.5mL centrifuge tube pipe is covered, draw of the aperture injection of 200 μ L preparations, make preparation cover mouse skin from 5mL centrifuge tube pipe lid with the 1mL syringe.Mice is placed the lucifuge place, and observe once every separated half an hour, and whether firm, whether seepage is arranged if observing the stickup of centrifuge tube and mouse skin, if any seepage leakages is pasted again, supplies preparation, and record.
2.4 bark fetching is observed:
After 6 hours,, in the new centrifuge tube of packing into, frozen in-80 ℃ with residue preparation sucking-off in the centrifuge tube.The centrifuge tube of mouse back is taken off, use the normal saline flushing mouse skin, each 1mL, flushing is at least 10 times altogether.Adopt disconnected neck method to put to death mice, carefully mention mouse skin,, absorb moisture, be stored in-80 ℃ along the vestige clip mouse skin that centrifuge tube is pasted with the ophthalmology tweezer, for use.
2.5 frozen section:
Make the frozen section of skin histology according to the operational approach of U.S. Leica company freezing microtome, the capable rip cutting thickness of skin is that 5 μ m are affixed on the microscope slide.
2.6 fluorescence microscope is observed down, and takes pictures.
Embodiment 2
The animal skin transdermal experiment
1. key instrument, reagent and material
1.1ICR mice: Nantong University's Experimental Animal Center, weigh 18～20g 4～6 ages in week, male and female are at random.
1.2 the siRNA of fluorescently-labeled siRNA:Cy3 labelling (Cy3-siRNA) (Biomics Bioisystech Co., Ltd).
1.3 instrument: freezing microtome (Leica CM1900, U.S. Leica company); Just putting fluorescence microscope (X3, Japanese Olympus company) etc.
1.4 reagent: 10% (m/V) chloral hydrate (Chemical Reagent Co., Ltd., Sinopharm Group); Medical glue (the medical glue company limited of Guangzhou white clouds); Sodium chloride injection (0.9%) (the healthy and free from worry pharmaceutcal corporation, Ltd in Shandong); (0.1MPBS Biomics Bioisystech Co., Ltd); Angelica essential oil (the natural medicinal oil company limited of Jiangxi Cedrus deoclar (Roxb.) G. Don); Azone (Aladdin reagent (China) company limited).
1.5 material: 1mL disposable syringe, 5mL disposable syringe (Jiangyan City newly revive medical apparatus and instruments company limited), disposable cotton swab, 5mL centrifuge tube, eye scissors, hospital gauze etc.
2. experimental procedure
2.1 the drug transdermal import system of preparation angelica essential oil mediation
2.1.1 the transdermal enhancer method for preparing is with step 2.1 among the embodiment 1
2.1.2 4mg siRNA is dissolved in the PBS solution of 0.1M that 10mL contains 25% glycerol, and mixing is subsequent use.
2.2 transdermal
Mice is divided into 3 groups at random, every group of 2 mices:
Table 1
Operating procedure is carried out with step 2.2-2.6 among the embodiment 1.
3. experimental result
As shown in Figure 2, for different percutaneous introduction agenies carry out the effect contrast figure of nucleic acid transdermal, white portion is the distribution of siRNA in cortex of Cy3 labelling among the figure, and wherein, a, b are the PBS negative control group, and c, d are the short group of passing through of angelica essential oil, and e, f are the short group of passing through of azone.Can be known that by Fig. 2 the siRNA of PBS negative control group is deposited on skin surface, the short group of passing through of short group thoroughly of angelica essential oil and azone has obvious siRNA entering dermal layer of the skin.
Embodiment 3
The experiment of percutaneous introduction agent nucleic acid molecules transdermal concentration dependent
1. key instrument, reagent and material
1.1ICR mice: Nantong University's Experimental Animal Center, weigh 18～20g 4～6 ages in week, male and female are at random.
1.2 the siRNA of fluorescently-labeled siRNA:Cy3 labelling (Cy3-siRNA) (Biomics Bioisystech Co., Ltd).
1.3 instrument: freezing microtome (Leica CM1900, U.S. Leica company); Just putting fluorescence microscope (X3, Japanese Olympus company) etc.
1.4 reagent: 10% (m/V) chloral hydrate (Chemical Reagent Co., Ltd., Sinopharm Group); Sodium chloride injection (0.9%) (the healthy and free from worry pharmaceutcal corporation, Ltd in Shandong); 0.1M PBS (Biomics Bioisystech Co., Ltd); Angelica essential oil (the natural medicinal oil company limited of Jiangxi Cedrus deoclar (Roxb.) G. Don); Jojoba oil (the gloomy biological product company limited of Zibo woods) etc.
1.5 material: 1mL disposable syringe, 5mL disposable syringe (Jiangyan City newly revive medical apparatus and instruments company limited), disposable cotton swab, 5mL centrifuge tube, eye scissors, hospital gauze etc.
2. experimental procedure
2.1 the drug transdermal import system of preparation angelica essential oil and Jojoba oil mediation
2.1.1 the transdermal enhancer method for preparing is with step 2.1 among the embodiment 1, difference is, with the mixture replacement angelica essential oil of angelica essential oil and Jojoba oil.
2.1.2 the method for preparing of nucleic acid preparation is with step 2.1.2 among the embodiment 2
2.2 transdermal
Mice is divided into 6 groups at random, and each handles 2 mices, and as base oil, angelica essential oil concentration is 10%, 20%, 50% and 100% compound essential Oil with Jojoba oil:
Table 2
Operating procedure is with step 2.2 among the embodiment 2.
3. experimental result
As shown in Figure 3, be the effect contrast figure that the angelica essential oil and the Jojoba oil compound prescription preparation of variable concentrations carries out the nucleic acid transdermal, white portion is the distribution of siRNA in cortex of Cy3 labelling among the figure.Wherein, A, b are the PBS negative control group; C, d are the short matched group that passes through of 100% Jojoba oil, and e, f are the short group of passing through of 10% angelica essential oil+90% Jojoba oil, and g, h are the short group of passing through of 20% angelica essential oil+80% Jojoba oil; I, j are the short group of passing through of 50% angelica essential oil+50% Jojoba oil, and k, l are the short group of passing through of 100% angelica essential oil.Can know by Fig. 3; The siRNA of PBS negative control group is deposited on skin surface; Short matched group and the PBS negative control group indifference of passing through of 100% Jojoba oil; Along with the raising of angelica essential oil concentration, the degree of depth that siRNA gets into skin increases, and explains that penetrating of siRNA has concentration dependent to angelica essential oil simultaneously.
Embodiment 4
Compound percutaneous imported agent cooperates nucleic acid emulsion, cream frost preparation to carry out transdermal experiment
1. key instrument, reagent and material
1.1ICR mice: Nantong University's Experimental Animal Center, weigh 18～20g 4～6 ages in week, male and female are at random.
1.2 contain the fluorescently-labeled siRNA cosmetic formulations of 0.2mg/mL:
1.2.1 nucleic acid emulsion I prescription
Table 3
1.2.2 nucleic acid cream frost I prescription
Table 4
| Sequence number | Component | Weight |
| 1 | Ammonium acryloyldime-thyltaurate// VP copolymer | 10g |
| 2 | siRNA | 0.02g |
| 3 | Essence | 0.5g |
| 4 | Antiseptic | 0.2g |
| 5 | Deionized water | Complement to 100g |
1.3 compound percutaneous imported agent (%, by weight percentage)
Table 5
| Numbering | Jojoba oil | Angelica essential oil | Flos Rosae Rugosae quintessence oil | Herba Lysimachiae foenum-graeci quintessence oil | The Olibanum quintessence oil | Tea tree ethereal oil |
| BW-01 | 96.9 | 2 | 0.2 | 0.3 | 0.5 | 0.1 |
| BW-02 | 62 | 20 | 6 | 5 | 3 | 4 |
| BW-03 | 38.4 | 50 | 0.7 | 0.6 | 10 | 0.3 |
1.4 instrument: freezing microtome (Leica CM1900, U.S. Leica company); Just putting fluorescence microscope (X3, Japanese Olympus company) etc.
1.5 reagent: 10% (m/V) chloral hydrate (Chemical Reagent Co., Ltd., Sinopharm Group); Sodium chloride injection (0.9%) (the healthy and free from worry pharmaceutcal corporation, Ltd in Shandong); 0.1M PBS (Biomics Bioisystech Co., Ltd) etc.
1.6 material: 1mL disposable syringe, 5mL disposable syringe (Jiangyan City newly revive medical apparatus and instruments company limited), disposable cotton swab, 5mL centrifuge tube, eye scissors, hospital gauze etc.
2. experimental procedure
2.1 the drug transdermal import system of preparation angelica essential oil and Jojoba oil mediation
2.1.1 the transdermal enhancer method for preparing is with step 2.1 among the embodiment 1, difference is, replaces angelica essential oil with the proportioning mixture in 1.3.
2.1.2 preparation nucleic acid preparation
2.1.2.1 preparation nucleic acid emulsion I
Respectively oil phase in the table 3 and water are heated to 85 ℃, and homogenizing, insulation 30min; Be cooled to 75 ℃, under stirring, oil phase be added to aqueous phase and stir; Stir the following 75 ℃ of homogenizing 3min of vacuum fast; Be cooled to 45 ℃ of sequence numbers in the adding table 3 and be 17,18,19 component, stir 20min, be cooled to room temperature.
2.1.2.2 preparation nucleic acid cream frost I
Deionized water is heated to 85 ℃, and constant temperature 30min postcooling to 45 ℃ stirs sequence number in the adding table 4 down and is 1,2,3,4 component, stirs into paste.
2.2 transdermal
Mice is divided into 8 groups at random, and each handles 2 mices:
Table 6
| Group | Percutaneous introduction agent | Nucleic acid preparation |
| 1 | PBS solution | Nucleic acid emulsion formulations I |
| 2 | BW-01 | Nucleic acid emulsion formulations I |
| 3 | BW-02 | Nucleic acid emulsion formulations I |
| 4 | BW-03 | Nucleic acid emulsion formulations I |
| 5 | PBS solution | Nucleic acid cream frost formula I |
| 6 | BW-01 | Nucleic acid cream frost formula I |
| 7 | BW-02 | Nucleic acid cream frost formula I |
| 8 | BW-03 | Nucleic acid cream frost formula I |
Inject 80 μ L, 10% chloral hydrate at each group ICR mouse peritoneal and anaesthetize, treat the mice holonarcosis after, mice is lain on the hospital gauze, use the eye scissors lagging to wipe out the about 1.5cm * 1.5cm of mouse back center hair, can not break mouse skin.At the bottom of wiping out the centrifuge tube pipe along the centrifugal mouth of pipe 1cm of 5mL place, take the centrifuge tube mouth of pipe one end, and the edge is repaired smooth.Draw the medical glue of 10uL and be coated on 5mL centrifuge tube oral area edge, then centrifuge tube is pressed on the mouse back skin, and lasting 10s.
Pass through 2.3 percutaneous introduction agent is short:
Dip in cotton swab and to get percutaneous introduction agent and evenly spread upon on the mouse skin in the centrifuge tube, and continue to smear 1min, repeat behind the 10min to smear once, smear altogether 3 times with cotton swab.
Smear for the third time and finish back 10min at interval, be coated with the position of percutaneous introduction agent, the remaining percutaneous introduction agent of removal skin surface with clean cotton swab wiping.Be coated with clean cotton swab and contain the fluorescently-labeled siRNA emulsion of 0.2mg/ml in right amount, smear evenly in skin of back.After mice is placed the lucifuge place.
2.4 bark fetching is observed:
After 6 hours,, in the new centrifuge tube of packing into, frozen in-80 ℃ with residue preparation sucking-off in the centrifuge tube.The centrifuge tube of mouse back is taken off, use the normal saline flushing mouse skin, each 1mL, flushing is at least 10 times altogether.Adopt disconnected neck method to put to death mice, carefully mention mouse skin,, absorb moisture, be stored in-80 ℃ along the vestige clip mouse skin that centrifuge tube is pasted with the ophthalmology tweezer, for use.
2.5 frozen section:
Make the frozen section of skin histology according to the operational approach of U.S. Leica company freezing microtome, the capable rip cutting thickness of skin is that 5 μ m are affixed on the microscope slide.
2.6 fluorescence microscope is observed down, and takes pictures.
3. experimental result
Fig. 4 carries out the effect contrast figure of nucleic acid transdermal for compound percutaneous imported agent cooperates nucleic acid emulsion I, and wherein, a, b are the PBS negative control group, and c, d are the short group of passing through of BW-01, and e, f are the short group of passing through of BW-02, and g, h are the short group of passing through of BW-03.
Fig. 5 carries out the effect contrast figure of nucleic acid transdermal for compound percutaneous imported agent cooperates nucleic acid cream frost I, and wherein, a, b are the PBS negative control group, and c, d are the short group of passing through of BW-01, and e, f are the short group of passing through of BW-02, and g, h are the short group of passing through of BW-03.
Visible by Fig. 4-5, white portion is the distribution of siRNA in cortex of Cy3 labelling among the figure, and compound percutaneous imported agent BW-01, BW-02, BW-03 can both effectively promote the absorption of siRNA.Wherein, nucleic acid emulsion I promotes the degree of depth of nucleic acid transdermal to be superior to nucleic acid cream frost I.
Embodiment 5
Compound percutaneous imported agent cooperates nucleic acid emulsion, cream frost preparation to carry out transdermal experiment
1. key instrument, reagent and material
1.1ICR mice: Nantong University's Experimental Animal Center, weigh 18～20g 4～6 ages in week, male and female are at random.
1.2 contain the fluorescently-labeled siRNA cosmetic formulations of 0.2mg/mL:
1.2.1 nucleic acid emulsion I prescription is with the table 3 among the embodiment 4
1.2.2 nucleic acid cream frost II prescription
Table 7
1.3 compound percutaneous imported agent (%, by weight percentage)
Table 8
| Numbering | Jojoba oil | Angelica essential oil | Flos Rosae Rugosae quintessence oil | Herba Lysimachiae foenum-graeci quintessence oil | The Olibanum quintessence oil | Tea tree ethereal oil | The Herba Menthae quintessence oil |
| BW-04 | 96.3 | 2 | 0.2 | 0.3 | 0.5 | 0.1 | 0.5 |
| BW-05 | 98 | 1.5 | - | - | - | - | 0.5 |
| BW-06 | 98 | 1 | 0.24 | - | 0.2 | 0.3 | 0.3 |
1.4 instrument: freezing microtome (Leica CM1900, U.S. Leica company); Just putting fluorescence microscope (X3, Japanese Olympus company) etc.
1.5 reagent: 10% (m/V) chloral hydrate (Chemical Reagent Co., Ltd., Sinopharm Group); Sodium chloride injection (0.9%) (the healthy and free from worry pharmaceutcal corporation, Ltd in Shandong); 0.1M PBS (Biomics Bioisystech Co., Ltd).
1.6 material: 1mL disposable syringe, 5mL disposable syringe (Jiangyan City newly revive medical apparatus and instruments company limited), disposable cotton swab, 5mL centrifuge tube, eye scissors, hospital gauze etc.
2. experimental procedure
2.1 the drug transdermal import system of preparation angelica essential oil and Jojoba oil mediation
2.1.1 the transdermal enhancer method for preparing is with step 2.1 among the embodiment 1, difference is, replaces angelica essential oil with the proportioning mixture in 1.3.
2.1.2 preparation nucleic acid preparation
2.1.2.1 preparation nucleic acid emulsion I
Method for preparing is with step 2.1.2.1 among the embodiment 4
2.1.2.2 preparation nucleic acid cream frost II
Oil phase in the table 7 and water are not heated to 85 ℃, and homogenizing, insulation 30min; Be cooled to 75 ℃, under stirring, oil phase be added to aqueous phase and stir; Stir the following 75 ℃ of homogenizing 3min of vacuum fast; Be cooled to 45 ℃ of sequence numbers in the adding table 7 and be 18,19,20 component, stir 20min, be cooled to room temperature.
2.2 transdermal
Mice is divided into 8 groups at random, handles 2 mices for every group:
Table 9
| Group | Percutaneous introduction agent | Nucleic acid preparation |
| 1 | PBS solution | Nucleic acid emulsion I |
| 2 | BW-04 | Nucleic acid emulsion I |
| 3 | BW-05 | Nucleic acid emulsion I |
| 4 | BW-06 | Nucleic acid emulsion I |
| 5 | PBS | Nucleic acid cream frost II |
| 6 | BW-04 | Nucleic acid cream frost II |
| 7 | BW-05 | Nucleic acid cream frost II |
| 8 | BW-06 | Nucleic acid cream frost II |
Operating procedure is with step 2.2 among the embodiment 2
3. experimental result
Fig. 6 carries out the effect contrast figure of nucleic acid transdermal for compound percutaneous imported agent cooperates nucleic acid emulsion I, and wherein, a, b are the PBS negative control group, and c, d are the short group of passing through of BW-04, and e, f are the short group of passing through of BW-05, and g, h are the short group of passing through of BW-06.
Fig. 7 carries out the effect contrast figure of nucleic acid transdermal for compound percutaneous imported agent cooperates nucleic acid cream frost II, and wherein, a, b are the PBS negative control group, and c, d are the short group of passing through of BW-04, and e, f are the short group of passing through of BW-05, and g, h are the short group of passing through of BW-06.
Visible by Fig. 6-7, white portion is the distribution of siRNA in cortex of Cy3 labelling among the figure, and compound percutaneous imported agent BW-04, BW-05, BW-06 can both effectively promote the absorption of siRNA.Wherein, nucleic acid emulsion formulations I promotes the degree of depth of nucleic acid transdermal to be superior to nucleic acid cream frost formula I I.
Embodiment 6
The animal transdermal experiment of different percutaneous introduction agenies
1. key instrument, reagent and material
1.1ICR mice: Nantong University's Experimental Animal Center, weigh 18～20g 4～6 ages in week, male and female are at random.
1.2 contain the fluorescently-labeled siRNA emulsion of 0.2mg/mL: nucleic acid emulsion formulations I (with embodiment 11).
1.3 instrument: freezing microtome (Leica CM1900, U.S. Leica company); Just putting fluorescence microscope (X3, Japanese Olympus company) etc.
1.4 reagent: 10% (m/V) chloral hydrate (Chemical Reagent Co., Ltd., Sinopharm Group); Sodium chloride injection (0.9%) (the healthy and free from worry pharmaceutcal corporation, Ltd in Shandong); 0.1M PBS (Biomics Bioisystech Co., Ltd) etc.
1.5 percutaneous introduction agent: Oleum Folium Artemisiae Argyi, eucalyptus oil, Oleum Caryophylli, Oleum Cinnamomi, Oleum Terebinthinae, Oleum menthae, hot thin oil, angelica essential oil, Borneolum Syntheticum (5% Borneolum Syntheticum is dissolved in 75% ethanol), Mentholum (5% Mentholum is dissolved in 75% ethanol) (the natural medicinal oil company limited of Jiangxi Cedrus deoclar (Roxb.) G. Don).
1.6 material: 1mL disposable syringe, 5mL disposable syringe (Jiangyan City newly revive medical apparatus and instruments company limited), disposable cotton swab, 5mL centrifuge tube, eye scissors, hospital gauze etc.
2. experimental procedure
2.1 pretreatment:
The ICR mouse peritoneal is injected 80 μ L, 10% chloral hydrate and is anaesthetized, treat the mice holonarcosis after, mice is lain on the hospital gauze, use the eye scissors lagging to wipe out the about 1.5cm * 1.5cm of mouse back center hair, can not break mouse skin.
Pass through 2.2 percutaneous introduction agent is short:
Dip in cotton swab and to get imported agent (and contrast PBS solution) and evenly spread upon on the mouse skin in the centrifuge tube, and continue to smear 1min, repeat behind the 10min to smear once, smear altogether 3 times with cotton swab.
2.3 transdermal:
Smear for the third time and finish back 10min at interval, be coated with the position of imported agent (and contrast PBS solution), remove the remaining imported agent (and contrasting PBS solution) of skin surface with clean cotton swab wiping.Be coated with clean cotton swab and contain the fluorescently-labeled siRNA emulsion of 0.2mg/ml in right amount, smear evenly in skin of back.After mice is placed the lucifuge place.
2.4 bark fetching is observed:
Behind the transdermal 6 hours,, in the new centrifuge tube of packing into, frozen in-80 ℃ with residual nucleus acid supplement sucking-off in the centrifuge tube.The centrifuge tube of mouse back is taken off, use the normal saline flushing mouse skin, each 1mL, flushing is at least 10 times altogether.Adopt disconnected neck method to put to death mice, carefully mention mouse skin,, absorb moisture, be stored in-80 ℃ along the vestige clip mouse skin that centrifuge tube is pasted with the ophthalmology tweezer, for use.
2.5 frozen section:
Make the frozen section of skin histology according to the operational approach of U.S. Leica company freezing microtome, the capable rip cutting thickness of skin is that 5 μ m are affixed on the microscope slide.
2.6 fluorescence microscope is observed down, and takes pictures.
3. experimental result
As shown in Figure 8, for cooperating nucleic acid emulsion formulations I, different percutaneous introduction agenies carry out the effect contrast figure of nucleic acid transdermal, and white portion is the distribution of siRNA in cortex of Cy3 labelling among the figure; Wherein, a, b are the short group of passing through of Oleum Folium Artemisiae Argyi, and c, d are the short group of passing through of eucalyptus oil; E, f are the short group of passing through of Oleum Caryophylli, and g, h are the short group of passing through of Oleum Cinnamomi, and i, j are the short group of passing through of Oleum Terebinthinae; K, l are the short group of passing through of Oleum menthae, and m, n are the short group of passing through of hot thin oil, and o, p are the short group of passing through of angelica essential oil; Q, r are the short group of passing through of Borneolum Syntheticum, and s, t are the short group of passing through of Mentholum, and u, v are the PBS negative control group.Can be known that by Fig. 8 compound percutaneous imported agent can both effectively promote the absorption of siRNA, these ten kinds of percutaneous introduction agenies all have transdermal effect, wherein angelica essential oil, eucalyptus oil, and the short effect of passing through of Oleum menthae is better.
Embodiment 7
Compound prescription (percutaneous introduction agent+nucleic acid molecules/nucleic acid preparation) transdermal experiment
1. key instrument, reagent and material
1.1ICR mice: Nantong University's Experimental Animal Center, weigh 18～20g 4～6 ages in week, male and female are at random.
1.2 contain the fluorescently-labeled siRNA cosmetic formulations of 0.2mg/mL:
1.2.1 nucleic acid emulsion compound prescription
Table 10
1.2.2 nucleic acid cream frost compound prescription
Table 11
1.4 instrument: freezing microtome (Leica CM1900, U.S. Leica company); Just putting fluorescence microscope (X3, Japanese Olympus company) etc.
1.5 reagent: 10% (m/V) chloral hydrate (Chemical Reagent Co., Ltd., Sinopharm Group); Sodium chloride injection (0.9%) (the healthy and free from worry pharmaceutcal corporation, Ltd in Shandong); 0.1M PBS (Biomics Bioisystech Co., Ltd).
1.6 material: 1mL disposable syringe, 5mL disposable syringe (Jiangyan City newly revive medical apparatus and instruments company limited), disposable cotton swab, 5mL centrifuge tube, eye scissors, hospital gauze etc.
2. experimental procedure
Mice is divided into 4 groups at random, every group of 3 mices:
Table 12
| Group | Nucleic acid preparation |
| 1 | Compound recipe nucleic acid emulsion |
| 2 | Compound recipe nucleic acid cream frost |
| 3 | Nucleic acid emulsion I |
| 4 | Nucleic acid emulsion II |
2.1 preparation compound prescription transdermal import system
2.1.1 preparation compound recipe nucleic acid emulsion
Respectively oil phase in the table 10 and water are heated to 85 ℃, and homogenizing, insulation 30min; Be cooled to 75 ℃, under stirring, oil phase be added to aqueous phase and stir; Stir the following 75 ℃ of homogenizing 3min of vacuum fast; Be cooled to 45 ℃ of sequence numbers in the adding table 10 and be 18,19,20 component, stir 20min, be cooled to room temperature.
2.1.2 preparation compound recipe nucleic acid cream frost
Respectively oil phase in the table 11 and water are heated to 85 ℃, and homogenizing, insulation 30min; Be cooled to 75 ℃, under stirring, oil phase be added to aqueous phase and stir; Stir the following 75 ℃ of homogenizing 3min of vacuum fast; Be cooled to 45 ℃ of sequence numbers in the adding table 11 and be 18,19,20 component, stir 20min, be cooled to room temperature.
2.1.3 preparation nucleic acid emulsion I
Method for preparing is with step 2.1.2.1 among the embodiment 4
2.1.4 preparation nucleic acid emulsion II
Adopt the nucleic acid emulsion II of preparation among the embodiment 5
2.2 pretreatment:
The ICR mouse peritoneal is injected 80 μ L, 10% chloral hydrate and is anaesthetized, treat the mice holonarcosis after, mice is lain on the hospital gauze, use the eye scissors lagging to wipe out the about 1.5cm * 1.5cm of mouse back center hair, can not break mouse skin.
2.3 transdermal:
Be coated with clean cotton swab and contain nucleic acid emulsion compound prescription and nucleic acid cream frost compound prescription in right amount in skin of back; Smear evenly; Use nucleic acid emulsion formulations I (with embodiment 11) and nucleic acid emulsion formulations II (with embodiment 12) as negative control simultaneously, press the preceding method operation.After the processing mice is placed the lucifuge place.
2.4 bark fetching is observed:
Behind the transdermal 6 hours,, in the new centrifuge tube of packing into, frozen in-80 ℃ with residue preparation sucking-off in the centrifuge tube.The centrifuge tube of mouse back is taken off, use the normal saline flushing mouse skin, each 1mL, flushing is at least 10 times altogether.Adopt disconnected neck method to put to death mice, carefully mention mouse skin,, absorb moisture, be stored in-80 ℃ along the vestige clip mouse skin that centrifuge tube is pasted with the ophthalmology tweezer, for use.
2.5 frozen section:
Make the frozen section of skin histology according to the operational approach of U.S. Leica company freezing microtome, the capable rip cutting thickness of skin is that 5 μ m are affixed on the microscope slide.
2.6 fluorescence microscope is observed down, and takes pictures.
3. experimental result
As shown in Figure 9; Be the effect contrast figure of compound prescription (percutaneous introduction agent+nucleic acid molecules/nucleic acid preparation) transdermal, white portion is the distribution of siRNA in cortex of Cy3 labelling among the figure, wherein; A, b are nucleic acid emulsion compound prescription group; C, d are nucleic acid cream frost compound prescription group, and e, f are nucleic acid emulsion formulations I matched group, and g, h are nucleic acid emulsion formulations II matched group.Visible by Fig. 9, nucleic acid emulsion compound prescription and nucleic acid cream frost compound prescription can both be effectively with the siRNA transdermals.
Claims (25)
1. the drug transdermal import system of a plant extract mediation is characterized in that, comprising: the percutaneous introduction agent that 1) prepares with at least a plant extract; 2) biomacromolecule or its preparation.
2. the drug transdermal import system of plant extract mediation as claimed in claim 1 is characterized in that described plant extract is a plant volatile oil.
3. the drug transdermal import system of plant extract mediation as claimed in claim 2; It is characterized in that described plant volatile oil is any one or a few the mixture in Radix Angelicae Sinensis volatile oil, Oleum Folium Artemisiae Argyi, eucalyptus oil, Oleum Caryophylli, Oleum Cinnamomi, Oleum Terebinthinae and the hot thin oil.
4. the drug transdermal import system of plant extract mediation as claimed in claim 1 is characterized in that described plant extract is a terpenoid.
5. the drug transdermal import system of plant extract as claimed in claim 4 mediation is characterized in that, described terpenoid is any one or a few the mixture in Oleum menthae, Borneolum Syntheticum and the Mentholum.
6. the drug transdermal import system of plant extract mediation as claimed in claim 1; It is characterized in that described percutaneous introduction agent is become by the Jojoba oil of 0～98 weight portion, the Radix Angelicae Sinensis volatile oil of 1～100 weight portion, the Flos Rosae Rugosae quintessence oil of 0.2～6 weight portion, the Herba Lysimachiae foenum-graeci quintessence oil of 0～5 weight portion, the Olibanum quintessence oil of 0～10 weight portion, the tea tree ethereal oil of 0～4 weight portion and the peppermint essence line of oils of 0～0.5 weight portion.
7. the drug transdermal import system of plant extract mediation as claimed in claim 1 is characterized in that described biomacromolecule is nucleic acid molecules, protein molecule, peptide molecule or insulin molecule.
8. the drug transdermal import system of plant extract mediation as claimed in claim 7 is characterized in that described biomacromolecule is a nucleic acid molecules, and described biomacromolecule preparation is a nucleic acid preparation.
9. the drug transdermal import system of plant extract as claimed in claim 8 mediation is characterized in that, described nucleic acid preparation comprise nucleic acid molecules and can with the preparation of its compatibility.
10. the drug transdermal import system of plant extract mediation as claimed in claim 9 is characterized in that described can be lipid or alcohols with the preparation of nucleic acid molecules compatibility.
11. drug transdermal import system like any described plant extract mediation among the claim 7-10; It is characterized in that described nucleic acid molecules is functional r NA and the version that contains modification or analogies and DNA and contains the version of modification or any one or a few the mixture in the analogies.
12. the drug transdermal import system of plant extract as claimed in claim 11 mediation is characterized in that, described functional r NA is any one or a few the mixture in siRNA, miRNA, little part RNA and the RNA aptamers.
13. the drug transdermal import system of plant extract mediation as claimed in claim 8 is characterized in that described nucleic acid preparation is the cosmetic emulsion that comprises nucleic acid molecules, comprises following component by weight percentage:
Olivem?1000 2.5～4％；
Oliwax?IC 0.5～2％；
16 octadecanol 1～6%;
Polydimethylsiloxane 2～4%;
Hydrogenation Fructus Canarii albi oleic acid Octyl Nitrite 1.5～8%;
PEG-100 stearate 0.5～4%;
Stearic acid 0.5～3%;
C15-19 alkane 3～6%;
Hydrogenated polydecene 2～7%;
Butylated hydroxytoluene 0.1～0.3%;
Butanediol 5～7%;
Glycerol 8.5～12%;
Xanthan gum 0.2～0.35%;
Hyaluronic acid 0.01～0.05%;
EDTA-disodium 0.1～0.3%;
siRNA 0.001～0.02％；
Essence 0.1～0.5%;
Antiseptic 0～1%;
Deionized water complements to 100%.
14. the drug transdermal import system of plant extract mediation as claimed in claim 8 is characterized in that, described nucleic acid preparation is the cosmetics cream frost that comprises nucleic acid molecules, comprises following component by weight percentage:
Olivem?1000 3～4％；
Oliwax?IC 1～2％；
Stearic acid 1～3%;
16 octadecanol 1～6%;
PEG-100 stearate 2.5～3%;
Hydrogenated polydecene 2～7%;
Caprylic/capric triglyceride 2～6%;
Butyrospermum parkii fruit fat 1～4%;
Isopropyl palmitate 2～4%;
Hydrogenation Fructus Canarii albi oleic acid Octyl Nitrite 1～7%;
Butylated hydroxytoluene 0.2～0.25%;
Butanediol 5～7%;
Glycerol 8.5～12%;
Xanthan gum 0.25～0.35%;
Hyaluronic acid 0.01～0.05%;
EDTA-disodium 0.1～0.3%;
siRNA 0.001～0.02％；
Essence 0.1～0.5%;
Antiseptic 0～1%;
Deionized water complements to 100%.
15. the drug transdermal method of a plant extract mediation; It is characterized in that; Adopt the drug transdermal import system of any described plant extract mediation among the claim 1-14; Concrete steps are: 1) urge with said percutaneous introduction agent earlier, then said biomacromolecule or its preparation are carried out transdermal; Or 2) with behind said biomacromolecule or its preparation and the said percutaneous introduction agent mix homogeneously, process the compound prescription preparation, carry out transdermal.
16. the drug transdermal method of plant extract as claimed in claim 15 mediation is characterized in that said method 2) in the compound prescription preparation comprise following component by weight percentage:
Angelica essential oil 1～20%;
Olivem?1000 2.5～4％；
Oliwax?IC 0.5～2％；
16 octadecanol 1～6%;
Polydimethylsiloxane 2～4%;
Hydrogenation Fructus Canarii albi oleic acid Octyl Nitrite 1.5～8%;
PEG-100 stearate 0.5～4%;
Stearic acid 0.5～3%;
C15-19 alkane 3～6%;
Hydrogenated polydecene 2～7%;
Butylated hydroxytoluene 0.1～0.3%;
Butanediol 5～7%;
Glycerol 8.5～12%;
Xanthan gum 0.2～0.35%;
Hyaluronic acid 0.01～0.05%;
EDTA-disodium 0.1～0.3%;
siRNA 0.001～0.02％；
Essence 0.1～0.5%;
Antiseptic 0～1%;
Deionized water complements to 100%.
17. the drug transdermal method of plant extract as claimed in claim 15 mediation is characterized in that said method 2) in the compound prescription preparation comprise following component by weight percentage:
Angelica essential oil 1～20%;
Olivem?1000 3～4％；
Oliwax?IC 1～2％；
Stearic acid 1～3%;
16 octadecanol 1～6%;
PEG-100 stearate 2.5～3%;
Hydrogenated polydecene 2～7%;
Caprylic/capric triglyceride 2～6%;
Butyrospermum parkii fruit fat 1～4%;
Isopropyl palmitate 2～4%;
Hydrogenation Fructus Canarii albi oleic acid Octyl Nitrite 1～7%;
Butylated hydroxytoluene 0.2～0.25%;
Butanediol 5～7%;
Glycerol 8.5～12%;
Xanthan gum 0.25～0.35%;
Hyaluronic acid 0.01～0.05%;
EDTA-disodium 0.1～0.3%;
siRNA 0.001～0.02％；
Essence 0.1～0.5%;
Antiseptic 0～1%;
Deionized water complements to 100%.
18. the application of the drug transdermal import system of any described plant extract mediation in preparation skin health product among the claim 1-14.
19. the application of the drug transdermal import system of plant extract mediation as claimed in claim 18 in preparation skin health product is characterized in that described skin health product is cosmetics.
20. the application of the drug transdermal import system of plant extract mediation as claimed in claim 19 in preparation skin health product is characterized in that described cosmetics are the cosmetics of Pear Power spot-removing function.
21. the application of the drug transdermal import system of plant extract mediation as claimed in claim 20 in preparation skin health product is characterized in that described cosmetics are the whitening cosmetics that contains the siRNA that regulates the melanin related gene expression.
22. the application of the drug transdermal method of any described plant extract mediation in preparation skin health product in the claim 15～17.
23. the application of the drug transdermal method of plant extract mediation as claimed in claim 22 in preparation skin health product is characterized in that described skin health product is cosmetics.
24. the application of the drug transdermal method of plant extract mediation as claimed in claim 23 in preparation skin health product is characterized in that described cosmetics are the cosmetics of Pear Power spot-removing function.
25. the application of the drug transdermal method of plant extract mediation as claimed in claim 24 in preparation skin health product is characterized in that described cosmetics are the whitening cosmetics that contains the siRNA that regulates the melanin related gene expression.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012103026269A CN102793639A (en) | 2011-12-16 | 2012-08-23 | Plant extract-mediated drug transdermal introducing system and transdermal method thereof |
| PCT/CN2012/081306 WO2014029143A1 (en) | 2012-08-23 | 2012-09-12 | Plant extract-mediated drug transdermal introducing system and transdermal method thereof |
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201110423002.8 | 2011-12-16 | ||
| CN201110423002 | 2011-12-16 | ||
| CN2012103026269A CN102793639A (en) | 2011-12-16 | 2012-08-23 | Plant extract-mediated drug transdermal introducing system and transdermal method thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102793639A true CN102793639A (en) | 2012-11-28 |
Family
ID=50150850
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2012103026269A Pending CN102793639A (en) | 2011-12-16 | 2012-08-23 | Plant extract-mediated drug transdermal introducing system and transdermal method thereof |
Country Status (2)
| Country | Link |
|---|---|
| CN (1) | CN102793639A (en) |
| WO (1) | WO2014029143A1 (en) |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105287265A (en) * | 2015-12-01 | 2016-02-03 | 广州丹奇日用化工厂有限公司 | Traditional Chinese medicine volatile oil and application thereof in cosmetics |
| CN105560077A (en) * | 2015-12-31 | 2016-05-11 | 上海宇昂新材料科技有限公司 | Penetration enhancer for cosmetics |
| CN107823637A (en) * | 2017-11-29 | 2018-03-23 | 安徽中医药大学 | A kind of leech peptide albumen cream and its preparation method and application |
| CN109223666A (en) * | 2018-12-04 | 2019-01-18 | 范承功 | A kind of plant extracts with transdermal enhancing effect and its application in skin care item |
| WO2020093393A1 (en) | 2018-11-06 | 2020-05-14 | 万华化学集团股份有限公司 | Aqueous polyurethane functional mask substrate and application thereof |
| CN111743852A (en) * | 2019-03-29 | 2020-10-09 | 傅柏舜 | Composition for making macromolecular medicine pass through skin |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110464662A (en) * | 2019-09-04 | 2019-11-19 | 朱梅年 | A kind of freckle-removing beauty cream and preparation method thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101087583A (en) * | 2004-12-22 | 2007-12-12 | 吉莱特公司 | Reduction of hair growth with survivin inhibitors |
| WO2009039743A1 (en) * | 2007-09-20 | 2009-04-02 | Suzhou Ribo Life Science Co., Ltd | Sirnas useful for inhibiting expression of tyrosinase gene, the compositions comprising the sirnas and their uses |
| CN102178662A (en) * | 2011-04-27 | 2011-09-14 | 王义明 | Insulin carrier transdermal drug delivery preparation and preparation method thereof |
| CN102451463A (en) * | 2010-10-23 | 2012-05-16 | 广州军区广州总医院 | Naja naja atra analgesic peptide transdermal preparation |
-
2012
- 2012-08-23 CN CN2012103026269A patent/CN102793639A/en active Pending
- 2012-09-12 WO PCT/CN2012/081306 patent/WO2014029143A1/en active Application Filing
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101087583A (en) * | 2004-12-22 | 2007-12-12 | 吉莱特公司 | Reduction of hair growth with survivin inhibitors |
| WO2009039743A1 (en) * | 2007-09-20 | 2009-04-02 | Suzhou Ribo Life Science Co., Ltd | Sirnas useful for inhibiting expression of tyrosinase gene, the compositions comprising the sirnas and their uses |
| CN102451463A (en) * | 2010-10-23 | 2012-05-16 | 广州军区广州总医院 | Naja naja atra analgesic peptide transdermal preparation |
| CN102178662A (en) * | 2011-04-27 | 2011-09-14 | 王义明 | Insulin carrier transdermal drug delivery preparation and preparation method thereof |
Non-Patent Citations (2)
| Title |
|---|
| N. KANIKKANNAN等: "Transdermal iontophoretic delivery of bovine insulin and monomeric human insulin analogue", 《JOURNAL OF CONTROLLED RELEASE》 * |
| 李莉等: "中草药挥发油类透皮吸收促进作用研究进展", 《中国新药杂志》 * |
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105287265A (en) * | 2015-12-01 | 2016-02-03 | 广州丹奇日用化工厂有限公司 | Traditional Chinese medicine volatile oil and application thereof in cosmetics |
| CN105560077A (en) * | 2015-12-31 | 2016-05-11 | 上海宇昂新材料科技有限公司 | Penetration enhancer for cosmetics |
| CN105560077B (en) * | 2015-12-31 | 2018-12-18 | 上海宇昂水性新材料科技股份有限公司 | Transdermal enhancer used for cosmetic |
| CN107823637A (en) * | 2017-11-29 | 2018-03-23 | 安徽中医药大学 | A kind of leech peptide albumen cream and its preparation method and application |
| WO2020093393A1 (en) | 2018-11-06 | 2020-05-14 | 万华化学集团股份有限公司 | Aqueous polyurethane functional mask substrate and application thereof |
| US11964032B2 (en) | 2018-11-06 | 2024-04-23 | Wanhua Chemical Group Co., Ltd. | Aqueous polyurethane functional mask substrate and application thereof |
| CN109223666A (en) * | 2018-12-04 | 2019-01-18 | 范承功 | A kind of plant extracts with transdermal enhancing effect and its application in skin care item |
| CN109223666B (en) * | 2018-12-04 | 2021-05-25 | 施莱新研(杭州)生物科技有限公司 | Plant extract composition and application thereof in skin care products |
| CN111743852A (en) * | 2019-03-29 | 2020-10-09 | 傅柏舜 | Composition for making macromolecular medicine pass through skin |
| CN111743852B (en) * | 2019-03-29 | 2022-10-28 | 傅柏舜 | Composition for making macromolecular medicine pass through skin |
Also Published As
| Publication number | Publication date |
|---|---|
| WO2014029143A1 (en) | 2014-02-27 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102793639A (en) | Plant extract-mediated drug transdermal introducing system and transdermal method thereof | |
| KR101894229B1 (en) | Functional composition comprising deer antlers derived stem cell culture medium | |
| KR101885501B1 (en) | Functional composition comprising deer antlers derived stem cell culture medium | |
| CN108969429A (en) | A kind of anti-acne essence | |
| CN104523473B (en) | A kind of anti-acne microemulsion system and preparation method thereof | |
| CN104288129A (en) | Stem cell micro-needle patch for resisting wrinkles and removing freckles and preparation method thereof | |
| CN112587593B (en) | Composition for treating acne and preparation method thereof | |
| CN104800269B (en) | A kind of Chinese medicine external emulsifiable paste for treating psoriasis and preparation method thereof | |
| CN113230317A (en) | Transdermal drug delivery preparation for treating knee osteoarthritis and preparation method thereof | |
| CN104491003B (en) | Add taste Yupingfeng gelling agent and its preparation method and application | |
| CN110974861B (en) | Blumea balsamifera oil liposome | |
| US20150297505A1 (en) | Anti-aging cosmetic composition including lipid-soluble fraction of bee venom and methods for preparing the same | |
| JP5543329B2 (en) | Use of Vernonia extract | |
| CN106729444A (en) | A kind of gel breast of repairing skin wound surfaces and preparation method thereof | |
| CN115531473B (en) | Compound essential oil for treating skin diseases and pharmaceutical composition containing same | |
| CN103845280B (en) | 20(R)-ginsenoside Rg3 preparation for external use, preparing method thereof and applications thereof | |
| CN103372052B (en) | Traditional Chinese medicine compound preparation for treating leucoderma and preparation method thereof | |
| CN106265792B (en) | The medicine and its detection method of a kind of anti-skin photoage | |
| KR102368646B1 (en) | Atopy treatment lotion utilizing room temperature emulsifying process | |
| CN108096315A (en) | A kind of preparation method and preparation of a new generation's anti-acne compound medicine | |
| CN111298075B (en) | Traditional Chinese medicine composition and preparation method and application thereof | |
| CN109528917B (en) | Korean medicinal composition for treating rheumatoid arthritis and preparation method of cream thereof | |
| CN112315879A (en) | Biological factor eyelash nourishing liquid and preparation method thereof | |
| CN111557956A (en) | Application of ganoderma spore oil in treating skin burn and scald | |
| CN102000176B (en) | Medicine composition for treating burns and scalds and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| ASS | Succession or assignment of patent right |
Owner name: BAIAO KECHUANG INSTITUTE OF MOLECULAR MEDICINE (NA Free format text: FORMER OWNER: BIOMICS BIOTECHNOLOGIES CO., LTD. Effective date: 20140926 |
|
| C41 | Transfer of patent application or patent right or utility model | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20140926 Address after: 226016 Jiangsu city of Nantong Province Economic and Technological Development Zone No. 76 Chang Xing Lu Applicant after: Biomics Branch Institute of molecular medicine (Nantong) Co., Ltd. Address before: 226016 Jiangsu city of Nantong Province Economic and Technological Development Zone No. 76 Chang Xing Lu Applicant before: Biomics Biotechnologies Co., Ltd. |
|
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| CB02 | Change of applicant information |
Address after: 226016 Jiangsu city of Nantong Province Economic and Technological Development Zone No. 76 Chang Xing Lu Applicant after: Study on molecular medicine 100 Kechuang source (Nantong) Co., Ltd. Address before: 226016 Jiangsu city of Nantong Province Economic and Technological Development Zone No. 76 Chang Xing Lu Applicant before: Biomics Branch Institute of molecular medicine (Nantong) Co., Ltd. |
|
| COR | Change of bibliographic data | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20121128 |
|
| RJ01 | Rejection of invention patent application after publication |