CN104208173A - Chinese herbal medicine composition, nanometer emulsion and transdermal patch, and preparation thereof and purpose for traumatic injury therapy - Google Patents
Chinese herbal medicine composition, nanometer emulsion and transdermal patch, and preparation thereof and purpose for traumatic injury therapy Download PDFInfo
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Abstract
The invention relates to a Chinese herbal medicine composition and a preparation method thereof. The composition comprises water solvent extracts and oil solvent extracts of red flower, radix dipsaci, panax notoginseng, and rheum officinale. The invention also relates to a oil-in-water type or a water-in-oil type nanometer emulsion and a preparation method thereof, the nanometer emulsion comprises an aqueous phase, an organic phase, and a surfactant, wherein the aqueous phase comprises a Chinese-herbal-medicine water solvent extract and/or the organic phase comprises a Chinese-herbal-medicine an oil solvent extract. Besides, the invention relates to a transdermal patch including the nanometer emulsion. The Chinese herbal medicine composition can effectively treat traumatic injury, especially the bone injury. The nanometer emulsion and the patch can substantially promote the transdermal absorption of effective components of Chinese herbal medicine and improve the biological utilization degree and the curative effect of the medicine.
Description
Technical field
The present invention relates to be used for the treatment of the Chinese herbal medicine formula of bone injury, and a kind of percutaneous patch that can be applicable to the nano-emulsion of described Chinese herbal medicine formula and can be applicable to described nano-emulsion.The invention still further relates to preparation and the purposes of described Chinese herbal medicine formula, nano-emulsion and percutaneous patch.
Background technology
Traumatic damage, for example musculoskeletal injuries---fracture is one of common disease of outpatient service.The global sickness rate of adult's fracture is estimated to be about in annual every 1000 people 9.0 to 22.8 people.In view of fracture is old people's common disease, can expect due to aging, bone fracture bone fracture disease number of cases will continue to increase in the near future.The patient of fracture needs one period of hospital stays growing very much before being discharged from hospitals upon recovery.Between 1997 to 1998, the average stay of New York hip fracture fixation is 5 days, and the longest is 33 days; The average stay of the masculinity and femininity fracture patient of suffering from Switzerland in 1992 is respectively 13.3 days and 19.6 days, and always length of stay is over 1,000,000 days.Fracture, when reducing social productive forces, has increased health resources utilization and social economical burden.
Although nowadays surgical operation can effectively reset and fix, hospital and clinical staff are seldom paid close attention to postoperative agglutination.After the reduction of the fracture, skeleton is fixed by foundry goods or holder, and healing after this mainly depends on the recovery of self.Patient the while in hospital except pain and inflammation treatment often in unmanned nurse state.Although a lot of scientific research personnel go to find the method that promotes bone healing, comprise that biomaterial scaffolds, somatomedin, bone morphogenetic protein and biophysics stimulate, these intervening measures are not also accepted by conventional clinical practice.
Promote that the healing of fracture is one of key area of the traditional Chinese medical science.Nearly all traditional Chinese medical science all relates to the local application of cataplasm of tcm (or ointment) to the processing of fracture, such therapeutic modality has been used thousands of years by Chinese.Yet, the too diversification and conventionally comprise for example 6 kinds of multiple Chinese herbal medicine, more frequent more than ten kind of the formula of these medical herbs.Such formula makes Chinese herb compound because the better systems proof of the science support of famine based on relevant evidence and clinical data is difficult to worldwide be accepted.Therefore there is such demand in this area: empirical tests has the simplification Chinese herbal medicine formula of good efficacy, lower toxic and side effects.
The topical application of Chinese herbal medicine formula is for example treated and is had a wide range of applications in bone injury treatment at various diseases.In localized drug delivery, the transportation of the percutaneous of Chinese herbal medicine effective ingredients is the key factor that medicine is given full play to therapeutic effect.Yet in traditional local application's agent, some Chinese herbal medicine active component is such as because the reasons such as poorly water-soluble are difficult to by skin diffusion, thereby limited the bioavailability of Chinese herbal medicine and the effectiveness of local application.Thus, this area also needs a kind of drug-supplying system of effectively dermal delivery Chinese herbal medicine active component, to improve bioavailability and the effectiveness of the Chinese herbal medicine formula of local application.
Summary of the invention
For meeting one or more in above-mentioned wilderness demand, the application provide a kind of aqueous that comprises Chinese herbal medicine and oiliness extract herbal composite, can be used for dispersing or dissolving the nano-emulsion of this herbal composite, matrix type percutaneous patch and their preparation method and the purposes in the traumatic tissue for the treatment of or skeletal injury that can be used for sending this herbal composite.
In one aspect, the present invention relates to a kind of herbal composite, the aqueous solvent extraction thing that it comprises Flos Carthami (Carthami Flos), Radix Dipsaci (Dipsaci Radix), Radix Notoginseng (Notoginseng Rhizoma) and Radix Et Rhizoma Rhei (Rhei Rhizoma) and oil-based solvent extract and optionally, pharmaceutically acceptable carrier, and wherein said herbal composite does not comprise the extract from Fructus Gardeniae (Fructus Gardeniae) and Ramulus Sambuci Williamsii (Sambucus Williamsii).
In one embodiment, compositions of the present invention is comprised of following: the aqueous solvent extraction thing of Flos Carthami, Radix Dipsaci, Radix Notoginseng and Radix Et Rhizoma Rhei and oil-based solvent extract and optionally, pharmaceutically acceptable carrier.
In one embodiment, described aqueous solvent is that water and/or described oil-based solvent are alcohol, preferred alcohol, the ethanol water such as 95%.
In one embodiment, the weight ratio of Flos Carthami, Radix Dipsaci, Radix Notoginseng and Radix Et Rhizoma Rhei is (0.8-1.2): (1-1.5): (0.8-1.2): (1-2), and preferably 1: 1: 1: 1 to 1: 1.5: 1: 2 scope.Preferably, the weight ratio of described aqueous solvent extraction thing and described oil-based solvent extract is 10: 1 to 7: 1.More preferably, the weight ratio of described aqueous solvent extraction thing and described oil-based solvent extract is counted 30%: 3% to 42%: 6% to account for the percentage by weight of described compositions, more preferably 35%: 4.3% to 39.8%: 5%.
In one embodiment, compositions of the present invention can be prepared by following steps:
A) prepare Flos Carthami, Radix Dipsaci, Radix Notoginseng and Radix Et Rhizoma Rhei, optionally by its stripping and slicing;
B) step raw material a) is heated to the sufficiently long time under immersion and/or backflow jointly or respectively in aqueous solvent, to obtain aqueous solvent extraction thing, optionally gained aqueous solvent extraction thing is selected to filtration, concentrated and cryodesiccated processing;
C) by step raw material or step b a)) the extraction residue of gained soaks and/or the sufficiently long time of reflux in oil-based solvent, to obtain oil-based solvent extract, optionally gained oil-based solvent extract is selected to filtration, concentrated and cryodesiccated processing; With
D) the described aqueous solvent extraction thing and the described oil-based solvent extract that obtain are formulated as to compositions, wherein optionally by the described aqueous solvent extraction thing obtaining and described oil-based solvent extract and the combination of at least one pharmaceutically acceptable carrier.
In one embodiment, compositions of the present invention can be prepared by following steps:
A) prepare Flos Carthami, Radix Dipsaci, Radix Notoginseng and Radix Et Rhizoma Rhei, optionally by its stripping and slicing;
B) step raw material a) is heated to the sufficiently long time under immersion and/or backflow jointly or respectively in oil-based solvent, to obtain oil-based solvent extract, optionally gained oil-based solvent extract is selected to filtration, concentrated and cryodesiccated processing;
C) by step raw material or step b a)) the extraction residue of gained soaks and/or the sufficiently long time of reflux in aqueous solvent, to obtain aqueous solvent extraction thing, optionally gained aqueous solvent extraction thing is selected to filtration, concentrated and cryodesiccated processing; With
D) combination obtains described aqueous solvent extraction thing and described oil-based solvent extract, wherein optionally by the described aqueous solvent extraction thing obtaining and described oil-based solvent extract and the combination of at least one pharmaceutically acceptable carrier.
In one embodiment, compositions of the present invention can be the form of paste, ointment, nano-emulsion or percutaneous patch.
In one aspect, the present invention relates to a kind of oil-in-water or water-in-oil type nanoemulsion agent, it comprises water, organic facies and surfactant, the oil-based solvent extract that the aqueous solvent extraction thing that wherein said water comprises one or more Chinese herbal medicine materials and/or described organic facies comprise described one or more Chinese herbal medicine materials.
In one embodiment, nano-emulsion of the present invention is oil-in-water type nano-emulsion.
In one embodiment, described water or organic facies have the drop of 10-200nm, preferred 10-50nm; And/or described water accounts for the 60-90% of described nano-emulsion, preferred 75-85% by weight; And/or described organic facies accounts for the 2-20% of described nano-emulsion, preferred 2-8% by weight; And/or described surfactant accounts for the 4-40% of described nano-emulsion, preferred 10-22% by weight.
The type that depends on nano-emulsion, depends on that described decentralized photo is water or organic facies, and described decentralized photo is water droplet or the oil droplet of 10-200nm, preferred 10-50nm.
In one embodiment, in nano-emulsion of the present invention, the aqueous phase solvent that forms described water is water; And/or the oil phase solvent that wherein forms described oil phase is one or more combination of Sefsol-218, Capryol-90, triglyceride, Myritol-318, limonene, liquid paraffin, ethanol or its; And/or wherein said surfactant is to have scope at the non-ionic surface active agent of 8 to 18 HLB value, for example Labrasol, castor oil hydrogenated EL (Cremophor EL), polysorbas20, polysorbate60 and Tween 80, be preferably castor oil hydrogenated EL and polysorbas20.
In one embodiment, in nano-emulsion of the present invention, described aqueous solvent extraction thing and described oil-based solvent extract are respectively aqueous solvent extraction thing as described above and oil-based solvent extract.
In one embodiment, the content of described aqueous extract in nano-emulsion by weight (W/W) be 5-10%; The content of described oiliness extract in nano-emulsion by weight (W/W) is 22-30%.
In one aspect, the present invention relates to a kind of method of preparing oil-in-water of the present invention or water-in-oil type nanoemulsion agent, it can comprise the following steps:
1) utilize aqueous solvent extraction Chinese herbal medicine material to obtain aqueous solvent extraction thing, described aqueous solvent is preferably water;
2) utilize oil-based solvent to extract Chinese herbal medicine material to obtain oil-based solvent extract, described oil-based solvent is preferably alcohol, more preferably 95% ethanol;
3) by step 1) the aqueous solvent extraction thing that obtains is dissolved in aqueous phase solvent, to form the water that comprises described aqueous solvent extraction thing, and by step 2) the oil-based solvent extract that obtains is dissolved in oil phase solvent, to form the organic facies that comprises described oil-based solvent extract;
4) described water and described organic facies are mixed with surfactant, form the front mixture of homogenize;
5), with 12,000-30, the speed of 000rpm, by mixture homogenization 10-30 minute before described homogenize, forms described nano-emulsion.
In one aspect, the present invention relates to a kind of percutaneous patch, it comprises lining, hypothallus and peel ply, and described hypothallus comprises oil-in-water type of the present invention or water-in-oil type nanoemulsion agent and binding agent, and described nano-emulsion preferably accounts for the 1-25% of described hypothallus by weight; Wherein said binding agent is preferably selected from the group being comprised of Carbopol, carbopol U20, carbopol HV-805EG, sodium carboxymethyl cellulose, gelatin, polyacrylic acid sodium salt, polyacrylic acid sodium salt NP700, polyvinylpyrrolidone 10 and PVP K90 and combination thereof, and described binding agent is the combination of polyacrylic acid sodium salt, polyacrylic acid sodium salt NP700, carbopol HV-805EG and polyvinylpyrrolidone more preferably; And described binding agent preferably accounts for the 10-15% of described hypothallus by weight.
In one embodiment, described hypothallus further comprises one or more compositions of the group that is selected from cross-linking agent, wetting agent, timbering material or filler and pH controlling agent composition.
In one embodiment, nano-emulsion, the binding agent of 10-15%, the wetting agent of the cross-linking agent of 0.01-0.8%, 15-25%, the timbering material of 1-10% or the filler that wherein said hypothallus comprises 1-25% with the weighing scale of hypothallus and the pH controlling agent that pH is controlled to 5-6.
In one aspect, the application provides herbal composite of the present invention, nano-emulsion or the percutaneous patch purposes in the medicine for the preparation for the treatment of traumatic damage.
In one embodiment, described traumatic damage is tissue injury or bone injury, and traumatic musculoskeletal injuries for example, such as soft tissue injury or fracture.
Accompanying drawing explanation
The aqueous extract that Fig. 1 a shows Flos Carthami prepared in accordance with the present invention, Radix Dipsaci, Radix Notoginseng and Radix Et Rhizoma Rhei is on impact that in the RAW264.7 cell of LPS induction, NO produces.
The oiliness extract that Fig. 1 b shows Flos Carthami prepared in accordance with the present invention, Radix Dipsaci, Radix Notoginseng and Radix Et Rhizoma Rhei is on impact that in the RAW264.7 cell of LPS induction, NO produces.
The effect of the aqueous extract that Fig. 2 a shows Flos Carthami prepared in accordance with the present invention, Radix Dipsaci, Radix Notoginseng and Radix Et Rhizoma Rhei to UMR-106 osteoblast vigor and propagation.
The effect of the oiliness extract of Flos Carthami, Radix Dipsaci, Radix Notoginseng and Radix Et Rhizoma Rhei prepared by Fig. 2 b demonstration the present invention to UMR-106 osteoblast vigor and propagation.
The osseous tissue of Fig. 3 a-3f demonstration damage is treated after 6 weeks through herbal composite according to the present invention, and yield strength, yield strength acting, ultimate strength, ultimate strength acting, breakdown strength, breakdown strength acting are able to remarkable improvement.
Fig. 4 a is presented at the comparison of the diffusion that receives the Chinese herbal medicine paste based on percentage by weight measured in compartment and nanometer patch.
Fig. 4 b is presented at the comparison of the diffusion of the Chinese herbal medicine paste based on percentage by weight measured in porcine skin and nanometer patch.
Fig. 5 is the schematic diagram of matrix type percutaneous patch prepared in accordance with the present invention.
The specific embodiment
The following specific embodiment is more fully set forth the present invention and multiple additional advantage thereof, so that the present invention and multiple additional advantage of the present invention are all more clear.
One aspect of the present invention provides a kind of herbal composite, the aqueous solvent extraction thing that it comprises Flos Carthami, Radix Dipsaci, Radix Notoginseng and Radix Et Rhizoma Rhei and oil-based solvent extract and optionally, pharmaceutically acceptable carrier, and described compositions does not comprise the extract from Fructus Gardeniae and Ramulus Sambuci Williamsii.Further, herbal composite of the present invention is comprised of following: the aqueous solvent extraction thing of Flos Carthami, Radix Dipsaci, Radix Notoginseng and Radix Et Rhizoma Rhei and oil-based solvent extract and optionally, pharmaceutically acceptable carrier.
Above-mentioned aqueous solvent and oil-based solvent can be that this area is applicable to respectively extracting hydrophilic segment contained in Flos Carthami, Radix Dipsaci, Radix Notoginseng and Radix Et Rhizoma Rhei and the acceptable any aqueous solvent of pharmacy and any oil-based solvent of hydrophobic part, make aqueous and the oiliness composition of Chinese herbal medicine, comprise particularly effective ingredient, extracted fully.
At least for said herbal medicine, i.e. Flos Carthami, Radix Dipsaci, Radix Notoginseng and Radix Et Rhizoma Rhei, preferred aqueous solvent is water, more preferably distilled water; Preferred oil-based solvent is 95% ethanol water.
Preferably, the weight ratio of Flos Carthami, Radix Dipsaci, Radix Notoginseng and Radix Et Rhizoma Rhei Chinese herbal medicine material can be (0.8-1.2): (1-1.5): (0.8-1.2): (1-2), and more preferably 1: 1: 1: 1 to 1: 1.5: 1: 2; Preferably, aqueous solvent extraction thing in described herbal composite and the weight ratio of oil-based solvent extract are 10: 1 to 7: 1, more preferably, the weight ratio of described aqueous solvent extraction thing and described oil-based solvent extract is counted 40%: 4% to 35%: 5% to account for the percentage by weight of described compositions, is preferably 35%: 4.3% to 39.8%: 5%.
Herbal composite described in can local application, with effective treatment traumatic damage.Described traumatic damage can comprise tissue (for example soft tissue) damage or bone injury, and traumatic musculoskeletal injuries for example, such as fracture.
Described treatment promotes the effect of knitting based on described herbal composite, as confirmed by embodiment below, the aqueous solvent extraction thing of wherein said compositions and the effect of oil-based solvent extract cooperative compensating, after fracture occurs, development and the osteanagenesis of Effective Regulation and coordination inflammation, promote cortical bone reparation, promote the healing of the osseous tissue of damage.Medicinal herb components used in the present invention for example, after abundant extraction (embodiment 1), and its aqueous solvent extraction thing and oil-based solvent extract comprise respectively hydrophilic active principle and the hydrophobic active component of medicinal herb components.The active component synergism of two types, significantly promotes the healing (as embodiment 2 verifies) of traumatic bone tissue injury.The application finds surprisingly, and the selected Chinese herb compound of the present invention is not only affected in knitting effect along with the simplification of formula, and has further significantly improved biomechanical properties in bones intensity, as shown in Figure 3.Meanwhile, the extract of raw material Flos Carthami, Radix Dipsaci, Radix Notoginseng and the Radix Et Rhizoma Rhei of herbal composite of the present invention for organism for example the toxic and side effects of human body greatly reduce, can, in effective treatment disease, reduce the risk that organism is caused to adverse effect.In addition, it all has clear and definite chemical index in the art the selected medicinal herb components of the present invention, and this makes to carry out quality control and transdermal research to it.
The present invention provides the preparation method of said herbal medicine compositions on the other hand, and it comprises:
A) prepare Flos Carthami, Radix Dipsaci, Radix Notoginseng and Radix Et Rhizoma Rhei, optionally by its stripping and slicing;
B) step raw material a) is heated to the sufficiently long time under immersion and/or backflow jointly or respectively in aqueous solvent, to obtain aqueous solvent extraction thing, optionally gained aqueous solvent extraction thing is selected to filtration, concentrated and cryodesiccated processing;
C) by step raw material or step b a)) the extraction residue of gained soaks and/or the sufficiently long time of reflux in oil-based solvent, to obtain oil-based solvent extract, optionally gained oil-based solvent extract is selected to filtration, concentrated and cryodesiccated processing; With
D) merge described aqueous solvent extraction thing and the described oil-based solvent extract obtaining, wherein optionally by the described aqueous solvent extraction thing and described oil-based solvent extract and the acceptable excipient composition of at least one pharmacy that obtain;
Step b wherein) and step c) order commutative, the oil-based solvent that can first carry out Chinese herbal medicine extracts, then carries out aqueous solvent extraction.Yet preferred step is first to carry out aqueous solvent extraction.Filtration step can be used to remove solid residue.
In solvent, soak and/or reflux under time of heating should long enough, fully to extract medicinal herb components.Above-mentioned four kinds of herbal raw materials can for example soak for example at least 1 hour in distilled water at aqueous solvent; Reflux in distilled water, for example, at least about 2 hours/time, preferably 2 times, can fully extract aqueous extract.The residue that aqueous is extracted generally can oil-based solvent for example in 95% alcoholic solution reflux for example within least 2 hours, can fully extract oiliness extract.Determine that the needed extraction conditions of different herbal raw materials is within those skilled in the art's limit of power.
The step that wherein the described aqueous solvent extraction thing obtaining and described oil-based solvent extract is formulated as to compositions can be according to the different medicament administration mode of expection.For example, the aqueous extract obtaining and oiliness extract can mix simply, or are formulated as and are applicable to the dosage form that different approaches is used with the acceptable excipient composition of any applicable pharmacy.Dosage form for local application, the described aqueous solvent extraction thing and the described oil-based solvent extract that obtain also can be mixed with local application nano-emulsion as mentioned below, to improve the dermal delivery of active constituents of medicine, thus bioavailability and the therapeutic effect of raising medicine.
In one embodiment, described extract can be distinguished after filtration, concentrates, after lyophilization, mix with 50% alcoholic solution, obtains the paste for local application.
The present invention provides a kind of nano-emulsion on the other hand, and it comprises water, organic facies and surfactant, the oil-based solvent extract that the aqueous solvent extraction thing that wherein said water comprises Chinese herbal medicine material and/or described organic facies comprise Chinese herbal medicine material.Depend on kind such as active constituents of medicine, content etc., described nano-emulsion can adopt oil-in-water type or water-in-oil type, and disperseing to connect can be water or oil phase mutually.According to determination of laser light scattering, the decentralized photo of nano-emulsion of the present invention can have 10nm to the drop of 200nm (water droplet or oil droplet depend on the type of nano-emulsion), preferably 10-50nm.
Those skilled in the art can, according to the type of example emulsion, determine the content of the composition such as water, oil phase in nano-emulsion.For example, water can account for the 60-90% of described nano-emulsion, preferred 75-85% by weight; And/or organic facies can account for the 2-20% of described nano-emulsion, preferred 2-8% by weight; And/or surfactant can account for the 4-40% of described nano-emulsion, preferred 10-22% by weight.The aqueous phase solvent that forms described water can be the acceptable aqueous phase solvent of any applicable pharmacy in this area, for example water.
The oil phase solvent that forms described oil phase can be the acceptable oil phase solvent of any applicable pharmacy in this area, for example Sefsol-218, Capryol-90, triglyceride, Myritol-318, limonene (limonene), liquid paraffin (liquid paraffin), ethanol or its combination.
Can add surfactant to reduce the interfacial tension between continuous phase and decentralized photo, thereby form Emulsion.The type (being oil-in-water type or water-in-oil type) of the nano-emulsion that can form according to expectation is selected suitable surfactant.The surfactant that is conducive to form oil-in-water emulsion is to have scope at the non-ionic surface active agent of 8 to 18 HLB value (being hydrophile-lipophile balance value).The example of this type of surfactant comprises Labrasol, castor oil hydrogenated EL (Cremophor EL), polysorbas20, polysorbate60 and Tween 80, and wherein castor oil hydrogenated EL and polysorbas20 are more preferred; Described surfactant preferably accounts for the 4-40% of described nano-emulsion, more preferably 10-22% by weight.
Importantly, the extract component of Chinese herbal medicine is dissolved in aqueous phase solvent or oil phase solvent completely, to guarantee sufficient homogenize.The aqueous solvent extraction thing of Chinese herbal medicine and oil-based solvent extract can adopt as described above and other any applicable methods extract and obtain.The consumption of described extract and solvent can according to the purposes of concrete Chinese herbal medicine, its for the type, extraction ratio, the dissolubility of extract in aqueous phase solvent and oil phase solvent etc. of disease, described extract by those skilled in the art, easily determined.
In one embodiment, when adopting oil-based solvent for example alcohol extract such as ethanol, the characteristic of the oil-based solvent based on adopted, resulting oil-based solvent extract comprises water-soluble part and oily molten part.In further embodiment, in extracting method of the present invention, Chinese herbal medicine first carries out water extraction, then carry out oil-based solvent extraction, thereby in oil-based solvent leaching process, in Chinese herbal medicine, remaining water-soluble part and whole oily molten part are extracted in the lump, and this causes an oil-based solvent extract part is hydrophilic, and a part is oil loving.Water-soluble part in the oiliness extract obtaining will be included in due to its hydrophilic in the water of nano-emulsion of final formation.In one embodiment, the content of aqueous extract in nano-emulsion is 5-10wt.%; The content of oiliness extract in nano-emulsion is 22-30wt.%.
In one embodiment, nano-emulsion of the present invention, for comprise the aqueous solvent extraction thing of said herbal medicine compositions (Flos Carthami, Radix Dipsaci, Radix Notoginseng and Radix Et Rhizoma Rhei) in its water, comprises the oil-in-water type nano-emulsion of the oil-based solvent extract of said herbal medicine compositions in its oil phase.In further embodiment, the solvent that forms described water can be water; And/or the solvent that forms described oil phase can be Capryol-90.The composition of this nano-emulsion, such as water, oil phase, aqueous solvent extraction thing and oil-based solvent extract etc., all can adopt technical scheme referred to above.
In one embodiment, nano-emulsion of the present invention comprises 74.4-84.8wt.% water, 3.8-3.9wt.% organic facies and 11.4-21.7wt.% surfactant.
In a further embodiment, nano-emulsion of the present invention comprises 74.4-84.8wt.% water, and wherein said aqueous phase solvent is water; 3.8-3.9wt.% organic facies, wherein said organic facies solvent is Capryol-90; And 11.4-21.7wt.% polysorbas20 is as surfactant.In further embodiment, the content of the aqueous extract that described water comprises in described nano-emulsion is 5-10%, and the content of the oiliness extract comprising in described oil phase in described nano-emulsion is 22-30%.Described nano-emulsion provides the special benefit of sending Chinese herbal medicine aqueous extract and oiliness extract, has significantly improved the percutaneous transmitance of Chinese herbal medicine active component.
The present invention provides a kind of method of preparing above-mentioned nano-emulsion on the other hand, and it comprises:
1) utilize aqueous solvent extraction Chinese herbal medicine material to obtain aqueous solvent extraction thing, described aqueous solvent is preferably water;
2) utilize oil-based solvent to extract Chinese herbal medicine material to obtain oil-based solvent extract, described oil-based solvent is preferably alcohol, more preferably 95% ethanol;
3) by step 1) the aqueous solvent extraction thing that obtains is dissolved in aqueous phase solvent, to form the water that comprises described aqueous solvent extraction thing, and by step 2) the oil-based solvent extract that obtains is dissolved in oil phase solvent, to form the organic facies that comprises described oil-based solvent extract;
4) described water and described organic facies are mixed with surfactant, form the front mixture of homogenize;
5), with 12,000-30, the speed of 000rpm, by mixture homogenization 10-30 minute before described homogenize, forms described nano-emulsion.
In one embodiment, the aqueous solvent extraction thing of Chinese herbal medicine material and oil-based solvent extract (being hydrophilic segment and the hydrophobic part of described Chinese herbal medicine material) difference (fully) are dissolved in to the aqueous phase solvent of the described water of formation and form in the oil phase solvent of described oil phase, to obtain the water that comprises described aqueous solvent extraction thing and the organic facies that comprises described oil-based solvent extract.
Optionally, described aqueous solvent extraction thing and oil-based solvent extract are to have carried out filtration, the extract that concentrated and/or lyophilization is processed, optional, the powder of described extract for processing through lyophilization.
In one embodiment, the aqueous solvent extraction thing of described Chinese herbal medicine material and oil-based solvent extract are respectively aqueous solvent extraction thing and the oil-based solvent extract in herbal composite mentioned above, i.e. the aqueous solvent extraction thing of Flos Carthami, Radix Dipsaci, Radix Notoginseng and Radix Et Rhizoma Rhei and oil-based solvent extract.Described extract optionally after filtration, concentrated for example concentrating under reduced pressure and the cryodesiccated powder morphology that is treated to.
In an embodiment of preparing nano-emulsion, surfactant is first evenly mixed with obtained water, then mix the front mixture of acquisition homogenize with uniform oil phase (being organic facies).Homogenize afterwards can adopt mechanical homogenizer to carry out, and for example, with 12,000-30, the speed of 000rpm is carried out 10-30 minute, and preferably with 24,000-30, the speed of 000rpm is carried out 20-30 minute.
The nano-emulsion that homogenize obtains after processing can carry out further high pressure homogenizing process, further to improve its homogeneity.For this high pressure homogenizing process, pressure can be arranged on 200-800 bar, and homogenize period can be set to 2-10 time.
Nano-emulsion of the present invention is compared with traditional Chinese herbal medicine dosage form, as verified by the part of embodiment below, can improve significantly the percutaneous diffusion of active constituents of medicine comprehensively, thereby the bioavailability of raising medicine improves the therapeutic effect of traumatic damage.And nano-emulsion of the present invention can comprise respectively multi-medicament active component in water and organic facies.
Nano-emulsion of the present invention is applicable to any Chinese herbal medicine formula that contains hydrophilic effective active composition and/or hydrophobicity effective active composition, herbal composite especially of the present invention.
The present invention provides a kind of percutaneous patch on the other hand, and it comprises lining, hypothallus and peel ply.
Wherein, described hypothallus comprises oil-in-water type or water-in-oil type nanoemulsion agent and binding agent as described above; Wherein said nano-emulsion preferably accounts for the 1-25% of described hypothallus by weight; The group that wherein said binding agent selects free Carbopol, carbopol U20, carbopol HV-805EG, sodium carboxymethyl cellulose, gelatin, polyacrylic acid sodium salt, polyacrylic acid sodium salt NP700, polyvinylpyrrolidone 10 and PVP K90 and combination thereof to form, described binding agent is preferably the combination of polyacrylic acid sodium salt, polyacrylic acid sodium salt NP700, carbopol HV-805EG and polyvinylpyrrolidone; And described binding agent preferably accounts for the 10-15% of described hypothallus by weight.
In one embodiment, described hypothallus further comprises one or more compositions that are selected from cross-linking agent, wetting agent, timbering material or filler and pH controlling agent.
In the substrate network of described hypothallus forms, tend to use cross-linking agent.The example of cross-linking agent comprises aluminum chloride, citric acid, dihydroxyaluminum aminoacetate (dihydroxyaluminum aminoacetate) and polyvinyl alcohol.For the hypothallus that comprises polyacrylic acid sodium salt, preferably use dihydroxyaluminum aminoacetate.Described cross-linking agent preferably accounts for the 0.01-0.8% of described hypothallus by weight.
In order to support also often to use timbering material or filler by described substrate network comprehensively.The example of described timbering material or filler comprises aluminium carbonate, Kaolin, silicon dioxide and zinc oxide.Wherein, preferably use Kaolin.Preferably, described timbering material accounts for the 1-10% of described hypothallus by weight.
In addition, described hypothallus also can comprise wetting agent to keep the moistening of patch.The example of described wetting agent comprises glycerol, propylene glycol, PEG-400, D-glucitol, Tween 80 and combination thereof.The wherein preferably combination of glycerol and D-glucitol.Preferably, described wetting agent accounts for the 10-40% of described hypothallus by weight, preferably 15-25%.
Described hypothallus also can further supplement pH controlling agent, for example tartaric acid.The pH of preferred described hypothallus is controlled in 5-6.
In one embodiment, nano-emulsion, the binding agent of 10-15%, the wetting agent of the cross-linking agent of 0.01-0.8%, 15-25%, the timbering material of 1-10% or the filler that described hypothallus comprises 1-25% with the weighing scale of substrate and the pH controlling agent that pH is controlled to 5-6.
In one embodiment, the nano-emulsion that hypothallus of the present invention comprises 17.4-18.4wt%, the binding agent of 12.4-12.7wt%, the wetting agent of the cross-linking agent of 0.4wt%, 19-19.7wt%, the timbering material of 4.6wt% or filler, the pH controlling agent that pH is controlled to 5-6 of 0.2wt% and the water of surplus.
In further embodiment, the nano-emulsion that hypothallus of the present invention comprises 17.4-18.4wt%; 8.1wt% polyacrylic acid sodium salt, 2.9wt% polyvinylpyrrolidone 10 and 1.4wt% carbopol U20 as binding agent or 8.1wt% polyacrylic acid sodium salt, 0.3wt% polyacrylic acid sodium salt NP-700,2.9wt% polyvinylpyrrolidone 10 and 1.4wt% carbopol HV-805EG as binding agent; 0.4wt% aluminum chloride or 0.4wt% dihydroxyaluminum aminoacetate are as cross-linking agent; 5.8wt% glycerol and 13.9wt%D-sorbitol are as wetting agent; 4.6wt% Kaolin is as timbering material; 0.2wt% tartaric acid is controlled at the pH controlling agent of 5-6 and the water of surplus as pH.In one aspect, the nano-emulsion that can be included in hypothallus of the present invention is defined nano-emulsion in the application.The percutaneous patch that comprises hypothallus of the present invention provides the special benefit of sending Chinese herbal medicine aqueous extract and oiliness extract, has significantly improved the percutaneous transmitance of Chinese herbal medicine (for example Flos Carthami, Radix Dipsaci, Radix Notoginseng and Radix Et Rhizoma Rhei and combination thereof) active component.Above-mentioned lining is the inertia lining providing support for patch of the present invention.Described peel ply covers and protects described hypothallus.
Lining can be based on poly film or allow different layers to be combined into the laminated material based on polyester (laminate) of independent meshi fabric.The example of lining comprises CoTran9720, CoTran9722, Scotchpak1109, Scotehpak9723, Scotchpak9730 and Scotchpak9735.The film based on polyester preferably not reacting with described hypothallus in the present invention.
Described peel ply preferably has low poisture-penetrability, and compatible with described hypothallus, to protect described hypothallus.
Percutaneous patch of the present invention can be prepared by following technique: the substrate or the substrate network that form described hypothallus are coated on described lining equably, are dried 6-24 hour at 65-100 ℃ in baking box, preferably dry 10-16 hour at 70-80 ℃; After cooling, peel ply is placed on described hypothallus to protect described hypothallus, obtains percutaneous patch.
The purposes of the percutaneous patch that the present invention relates on the one hand herbal composite as described above, the nano-emulsion that comprises it or comprises it (comprising the aqueous solvent extraction thing and the oil-in-water of oil-based solvent extract or the percutaneous patch of water-in-oil type nanoemulsion agent that contain described herbal composite) in the medicine for the preparation for the treatment of traumatic damage.Described purposes promotes the effect of knitting based on herbal composite of the present invention mentioned above.
In one embodiment, described traumatic damage comprises tissue (for example soft tissue) damage or bone injury; Traumatic musculoskeletal injuries for example, such as fracture.
Herbal composite and the described nano-emulsion that comprises it described in preferred local application.
Based on nano-emulsion of the present invention, promote the effect of the percutaneous diffusion transportation of active constituents of medicine, the aqueous solvent extraction thing of herbal composite of the present invention and the oil-in-water of oil-based solvent extract or water-in-oil type nanoemulsion agent and the percutaneous patch that comprises described nano-emulsion are when local application, the percutaneous diffusion of described extract Chinese medicine active component will be promoted, can improve the bioavailability of medicine, thereby further improve the therapeutic effect of traumatic damage.
Below with reference to specific embodiment, the present invention will be described in detail.Yet those skilled in the art understand, described specific embodiment only, in order more clearly to explain the present invention, in no case should be considered as any limitation of the invention.Unless stated otherwise, all commercially available acquisition and all content is all by weight for all reagent and equipment.
embodiment
Four taste Chinese herbal medicine Flos Carthamis, Radix Dipsaci, Radix Notoginseng and the Radix Et Rhizoma Rhei using in embodiment can, according to the method for recommending in Chinese Pharmacopoeia 2010 editions, adopt thin layer chromatography to identify.Flos Carthami, Radix Dipsaci, Radix Notoginseng and the Radix Et Rhizoma Rhei below adopting be all purchased from the Chinese medicine supplier of locality, Hong Kong, and according to said method, identified.
The aqueous solvent extraction thing that embodiment 1 preparation comprises Flos Carthami, Radix Dipsaci, Radix Notoginseng and Radix Et Rhizoma Rhei and the herbal composite of oil-based solvent extract
Prepare four kinds of medical herbs raw materials (weight ratio of Flos Carthami, Radix Dipsaci, Radix Notoginseng and Radix Et Rhizoma Rhei is 1: 1: 1: 1) totally 250 grams, be cut into segment, in 1.0L distilled water, soak 1 hour.Reflux boils 2 times, and each 2 hours, collect water extract, by a slice absorbent carbasus, filter.Residue boils 2 hours with the further reflux of 95% ethanol.Collect ethanol extraction absorbent carbasus filtration again.The filtrate of water and ethanol is concentrating under reduced pressure at 50 ℃ respectively, and then in lyophilization system (Freezone12, Labconco, Missouri, the U.S.), lyophilizing becomes powder.
The extraction ratio of water extract [CDNR (aq)] and ethanol extraction [CDNR (e)] is respectively 39.8%w/w and 5.0%w/w.
Get 19.5g CDNR (aq) and 3.0g CDNR (e) and be mixed with the Chinese herbal medicine paste for local application with 17ml50% ethanol.
The aqueous solvent extraction thing that embodiment 2 comprises Flos Carthami, Radix Dipsaci, Radix Notoginseng and Radix Et Rhizoma Rhei and the herbal composite of oil-based solvent extract are for the effect that promotes knitting
Mouse monokaryon cell/macrophage RAW254.7 and rat osteoblast UMR-106 are all purchased from US mode culture collection center (ATCC, the U.S.).In employing embodiment 1, Flos Carthami, Radix Dipsaci, Radix Notoginseng and the Radix Et Rhizoma Rhei aqueous extract (CDNR (aq)) of preparation and oiliness extract (CDNR (e)) are respectively as testing sample.
the oil-based solvent extract of Flos Carthami, Radix Dipsaci, Radix Notoginseng and Radix Et Rhizoma Rhei suppresses the RAW254.7 of LPS induction
nitric oxide production generation in cell
Anti-inflammatory (nitric oxide that is lipopolysaccharide (LPS) induction suppresses) for external test herbal composite of the present invention, employing contains 10% hyclone (Life Technologies, USA), the DMEM in high glucose (3500mg/L of 100U/ml penicillin and 100mg/L streptomycin (Life Technologies, USA); Purchased from Life Technologies, USA) culture medium, at 37 ℃, 5%CO
2cell culture incubator in cultivate mice RAW264.7 macrophage.Between culture period, regularly go down to posterity.
According to the difference of solubility properties, adopt respectively above-mentioned DMEM culture medium or DMSO aqueous solution (1%, v/v) dissolve CDNR (aq) to 0,50,100,200 and 400 μ g/mL, dissolve CDNR (e) to 0,25,50,100 and 200 μ g/mL; In this test, 1% DMSO aqueous solution produces and detects all without impact for Growth of Cells and NO.
With 4 * 10
6cells/well is inoculated in RAW264.7 cell in 24 orifice plates, is cultured to after cell attachment, adopts respectively the CDNR (aq) of variable concentrations and CDNR (e) to process cultured cell 24h; Each concentration for the treatment of arrange three parallel.Each hole adds 1 μ g/ml LPS afterwards, continues to cultivate 24h.Collect supernatant, add the Griess reagent of same volume, place 15min.Under 540nm, measure each hole absorbance, according to absorbance, calculate nitric oxide production growing amount.
As shown in Fig. 1 a and Fig. 1 b, in the RAW264.7 cell of LPS induction, CDNR (aq) does not affect nitric oxide output, and (Fig. 1 a); By contrast, in the RAW264.7 cell of LPS induction, compare with benchmark matched group (i.e. 0 concentration group), the CDNR (e) of 100 μ g/ml and 200 μ g/ml significantly suppresses respectively 51% and 77% (p < 0.05) (Fig. 1 b) of nitric oxide output.Therefore, the oil-based solvent extract of Flos Carthami, Radix Dipsaci, Radix Notoginseng and Radix Et Rhizoma Rhei has potent effect aspect antiinflammatory.
Adopt said method, with the Chinese herbal medicine paste of embodiment 1 preparation, test the mixture of aqueous extract and oiliness extract for the inhibition of nitric oxide output, obtain similarly suppressing result.This shows that aqueous extract does not affect the anti-inflammatory effect of oil-based solvent extract.
the aqueous solvent extraction thing of Flos Carthami, Radix Dipsaci, Radix Notoginseng and Radix Et Rhizoma Rhei promotes the work of UMR-106 cell
power and propagation
Adopt the DMEM in high glucose (3500mg/L of the hyclone (Life Technologies, USA), 100U/ml penicillin and the 100mg/L streptomycin (Life Technologies, USA) that contain 10%; Life Technologies, USA) culture medium, at 37 ℃, 5%CO
2cell culture incubator in cultivate rat bone sarcoma UMR-106 cell, between culture period, regularly go down to posterity.
According to the difference of solubility properties, (1%, v/v) dissolving is dissolved to medicine to be measured (being CDNR (aq) and CDNR (e)) respectively the concentration of 0,6.25,12.5,25,50 and 100 μ g/mL to adopt respectively culture medium or DMSO.
The impact of medicine on cell proliferation to be measured adopts BrdU-ELISA test kit (Roches, USA) to measure.With 1000 cells/well, UMR-106 cell is inoculated in 96 orifice plates, is cultured to after cell attachment, adopt respectively the medicine to be measured of variable concentrations and cultured cell jointly to hatch processing 24h; Each concentration for the treatment of arrange three parallel.Every hole adds BrdU working solution (forming with 6 μ L/mL dilutions by the storage liquid of the 5mg/ml) labeled cell of 10 μ L afterwards, continues to hatch 2h.Remove afterwards supernatant and with lavation buffer solution washing, add anti-BrdU antibody, hatch 120 minutes.According to the result of the step measurements cell proliferation providing in the appended description of test kit.
The effect of the medicine on cell proliferation to be measured of variable concentrations adopts the ratio of the cell proliferation result of administration group and blank solvent matched group (i.e. 0 concentration group) to represent (the cell proliferation result of blank solvent matched group counts 100%).
As shown in Figure 2 a and 2 b, UMR-106 osteoblast is processed 24 hours through the CDNR (aq) of 6.25-100 μ g/ml (p < 0.01), can be observed significant vigor strengthens and cultivation effect, the two respectively from 11% rising to 20% (Fig. 2 a), and rise to 22% (Fig. 2 b) from 6%.Yet UMR-106 osteoblast is being processed after 24 hours through CDNR (e), significantly improving does not appear in vigor and propagation, so this CDNR (e) composition does not play a role.Visible, the aqueous solvent extraction thing of Flos Carthami, Radix Dipsaci, Radix Notoginseng and Radix Et Rhizoma Rhei is promoting skeleton to have potent effect in forming.
Adopt said method, with the Chinese herbal medicine paste test aqueous extract of embodiment 1 preparation and the mixture of oiliness extract for
the vigor of UMR-106 cell and propagationeffect, obtain similarly promoting result.This shows that oiliness extract does not affect the effect of the promotion skeleton formation of aqueous solvent extraction thing.
in body, assessment herbal composite paste is to promoting the effect of knitting
20 monthly ages are that the female Sprague-Dawley rat of 15.2 ± 1.41 (means standard deviation) is purchased from the laboratory animal service centre of Hong Kong Chinese University (CUHK).All rats are raised in temperature control (25 ℃) and light-operated (circulation at daytime/night in 12 hours) environment.
First rat is anaesthetized by the mixture of intramuscular injection ketamine and xylazine (80mg/kg ketamine and 8mg/kg xylazine).Then on the femur of left side, the stage casing that electricity consumption is drilled in femur gets out two adjacent double-side-holes (each bore dia is 2mm) by A-P mode.With dentistry milling, bore two bridges are connected into the damaged of 2mm * 4mm.On the tibia of right side, electricity consumption is drilled in PM, and by interior-outer mode, to get out diameter be that the bilateral bone of 2.4mm is damaged.All boring procedures are all through 0.9% physiological saline solution perfusion sterilization, and boring is removed residual GUSUIPIAN with a large amount of normal saline washings before cut closure.
Rat is divided into two groups by 10 every group.In matched group, on the left side of rat femur and right side tibia, cover self-adhesive plastic film and do not do any treatment; In Chinese herbal medicine paste treatment group, on the left side of rat femur and right side tibia, the paste of preparation in the embodiment 1 of local application 0.5ml, covers described dextrin with self-adhesive plastic film and comes off and become dry to prevent it.The whole treatment cycle time is 6 weeks, during within every 2 days, change all thin film and paste.At the 42nd day, rat is implemented to euthanasia.Collect the left side of rat and the Thigh bone on right side, remove unnecessary soft tissue but retain periosteum.Use Hounsfield material testing machine (KM25, Redhill, Britain) to carry out four-point bending experiment.Load the load unit of maximum 2500N, the span of upper and lower supporting member is respectively 8.0 and 20mm.By the damaged centre that is positioned over two upper supporting pieces of the boring bone that is positioned at femur stage casing, then with the constant speed of mode 5mm/min from back to front, to sample, exert pressure until rupture.Record loads surrender (load at yield) for analyzing.All data of the femur of holing (left side) are all normalized with normal femur (right side).Its result is expressed as take the normalized ratio that normal femur is benchmark.
As shown in Fig. 3 a-3f, Chinese herbal medicine paste composition of the present invention has significantly promoted the biomechanics characteristic of bone in knitting process.In the cortex femur of boring, compare with matched group, after treatment in 42 days, paste treatment group shows to be increased by 15% normalized yield strength and increases by 13% surrender merit (P < 0.05) (Fig. 3 a and 3b).Paste treatment group has also shown beneficial effect in ultimate strength (Fig. 3 c and 3d) and breakdown strength (Fig. 3 e and 3f).Four-point bending is tested and is shown, herbal composite of the present invention can significantly improve the damaged biomechanical property (bone strength) of boring bone in healing.
According to the above results, visible, the nitric oxide that the herbal composite of the present invention of the ethanol extraction that comprises Flos Carthami, Radix Dipsaci, Radix Notoginseng and Radix Et Rhizoma Rhei can significantly suppress LPS induction produces, thus inflammation-inhibiting." antiinflammatory " is a key Therapeutic Principle for the treatment of fracture in theory of Chinese medical science, and it is intended to control and alleviate swelling and the pain of fracture site and surrounding soft tissue.On the other hand, the herbal composite of the present invention of the water extract that comprises Flos Carthami, Radix Dipsaci, Radix Notoginseng and Radix Et Rhizoma Rhei can significantly promote vigor and the propagation of UMR-106 cell, thereby promotion osteoblast regeneration, and osteoblast regeneration is the important mechanisms of bone repair process.This is at union of fracture repairing phase, when endochondral ossification occurs and osteoblast starts when cartilage callus forms new stratiform bone especially effective.
The biomechanics test result of above-mentioned in vivo test provides local application herbal composite of the present invention to promoting the strong evidence of knitting.Compared with the control, the counter-bending ability of higher femur that herbal composite of the present invention produces shows, applies its treatment and can improve before bone strength to bear more multiple pressure power in generation permanent deformation (may be caused by micro-fracture).Above-mentioned the results show, the aqueous solvent extraction thing that the present invention comprises Flos Carthami, Radix Dipsaci, Radix Notoginseng and Radix Et Rhizoma Rhei and the herbal composite of oil-based solvent can be by external approach Effective Regulation inflammatory and osteanagenesis, and in body, experiment has also confirmed that it is in the effect promoting aspect cortical bone reparation.The paste of directly using described the present embodiment has obtained similar effect on the skin of fracture time.
Adopt other ratio (0.8: 1: 0.8: 1, 1.2: 1.5: 1.2: 2 and 1: 1.5: 1: Flos Carthami 2), Radix Dipsaci, the Chinese herbal medicine paste of the aqueous solvent extraction thing of Radix Notoginseng and Radix Et Rhizoma Rhei compositions and the preparation of oil-based solvent extract, the percentage by weight that wherein aqueous solvent extraction thing and oil-based solvent extract account for respectively Chinese herbal medicine paste changed between 30%: 3% to 42%: 6%, comprise 35%: 4.3% to 39.8%: 5%), repeat above-mentioned test, obtain similarly regulation and control inflammatory and osteanagenesis, promote the significant effects (data are not shown) that promote traumatic bone healing and organization healing such as cortical bone reparation.
Toxicity test shows, Flos Carthami of the present invention, Radix Dipsaci, Radix Notoginseng and Radix Et Rhizoma Rhei Chinese herbal medicine formula also reduce (data are not shown) greatly to the toxicity of organism.
The preparation of the oil-in-water type nano-emulsion that embodiment 3.1 comprises Chinese herbal medicine aqueous extract and oiliness extract
According to the method for embodiment 1, prepare respectively aqueous extract powder and the oiliness extract powder of Chinese herbal medicine material Flos Carthami, Radix Dipsaci, Radix Notoginseng and Radix Et Rhizoma Rhei.
Take respectively the obtained about 1g of aqueous extract powder, the about 4g of oiliness extract powder, the about 1g of Capryol-90 and the about 7g of water; Mix homogeneously; Supersound process is so that whole extract powders fully dissolves.Under room temperature, standing making is separated; Collect upper strata and obtain organic facies, bottom is water.
Get 1.71g surfactant polysorbas20 and first mix with 12.73g water, gentle agitation under magnetic stirring apparatus, mix homogeneously.Mix with 0.56g organic facies afterwards, supersound process 5min, obtains pre-homogenize mixture.
Adopt mechanical homogenizer with the rotary speed of 24000rpm by described pre-homogenize mixture homogenization 20min, form nano-emulsion of the present invention.
Through laser light scattering (90Plus/BI-MAS, Brooker Hai Wen (the 90Plus/BI-MAS of instrument company, Brookhaven Instruments Corporation), the U.S.) measure, in formed nanoemulsions, the average diameter of contained oil droplet is 200nm, and it contains 3.8wt.% organic facies, 84.8wt.% water and 11.4wt.% surfactant.
The preparation of the oil-in-water type nano-emulsion that embodiment 3.2 comprises Chinese herbal medicine aqueous extract and oiliness extract
According to the method for embodiment 3.1, obtain and comprise respectively the aqueous extract of Flos Carthami, Radix Dipsaci, Radix Notoginseng and Radix Et Rhizoma Rhei and the water of oiliness extract and oil phase.
First polysorbas20 3.48g is mixed to gentle agitation under magnetic stirring apparatus, mix homogeneously with 11.9g water.Mix with 0.62g organic facies afterwards, supersound process 5min, obtains pre-homogenize mixture.
Adopt mechanical homogenizer with the rotary speed of 30000rpm by described pre-homogenize mixture homogenization 30min, form nano-emulsion of the present invention.
Through determination of laser light scattering, in formed nanoemulsions, the average diameter of contained oil droplet is 40nm, and it contains 3.9wt.% organic facies, 74.4wt.% water and 21.7wt.% surfactant (polysorbas20).
The preparation of the percutaneous patch that embodiment 4.1 comprises nano-emulsion of the present invention.
Mix 7g polyacrylic acid sodium salt, 4g Kaolin, 0.35g aluminum chloride, 5g glycerol and 12g D-glucitol and obtain A phase.0.15g tartaric acid is added and in 15g water, obtains 0.1wt.% tartaric acid solution; Then 2.5g polyvinylpyrrolidone 10 is dissolved in and in this tartaric acid solution, obtains B phase.Under the high-speed stirred of magnetic stir bar, by 1.2g carbopol U20 swelling in 24g water, obtain colloidal liquid, i.e. C phase.
First B phase is mixed mutually with C, under 1400rpm speed, stir 3min, mix mutually with A afterwards.The mixture solution (being the mixture of A phase, B phase and C phase) obtaining is sieved to filtration by 0.5mm, and the nano-emulsion obtaining with the embodiment 3.1 of 15g mixes, and under 800rpm, stirs 10min, obtains mixture, is used to form the substrate of hypothallus.
The water that the substrate obtaining contains 8.1wt% polyacrylic acid sodium salt, 4.6wt% Kaolin, 0.4wt% aluminum chloride, 5.8wt% glycerol, 13.9wt%D-sorbitol, 0.2wt% tartaric acid, 2.9wt% polyvinylpyrrolidone 10,1.4wt% carbopol U20,17.4wt% nano-emulsion and surplus.
Obtained substrate is coated on Scotchpak9735 lining equably with any applicable operation in this area, at 75 ℃, toasts 12 hours.After cooling, Scotchpak1022 peel ply is placed on to obtained sticking, is placed on hypothallus to protect this hypothallus, finally obtain patch.
The thickness of this patch is 1.1mm, and bonding adhesive attraction is 0.33N.
The preparation of the percutaneous patch that embodiment 4.2 comprises nano-emulsion of the present invention.
Mix 7g polyacrylic acid sodium salt, 0.25g polyacrylic acid sodium salt NP-700,4g Kaolin, 0.35g dihydroxyaluminum aminoacetate and 12g D-glucitol and obtain A phase.0.15g tartaric acid is added and in 15g water, obtains 0.1wt.% tartaric acid solution; Then 2.5g polyvinylpyrrolidone 10 is dissolved in and in this tartaric acid solution, obtains B phase.Under the high-speed stirred of magnetic stir bar, by 1.2g carbopol HV-805EG swelling in 24g water, obtain colloidal liquid, i.e. C phase.
First B phase is mixed mutually with C, under 1400rpm speed, stir 3min, mix mutually with A afterwards.The mixture solution (being the mixture of A phase, B phase and C phase) obtaining is sieved to filtration by 0.5mm, the nano-emulsion obtaining with the embodiment 3.2 of 4.5g glycerol and 16g mixes, under 800rpm, stir 10min, obtain mixture, be used to form the substrate of hypothallus.
The water that the substrate obtaining contains 8.1wt% polyacrylic acid sodium salt, 0.3wt% polyacrylic acid sodium salt NP-700,4.6wt% Kaolin, 0.4wt% dihydroxyaluminum aminoacetate, 13.8wt%D-sorbitol, 0.2wt% tartaric acid, 2.9wt% polyvinylpyrrolidone 10,1.4wt% carbopol HV-805EG, 5.2wt% glycerol, 18.4wt% nano-emulsion and surplus.
Obtained substrate is coated on Scotchpak9735 lining equably with any applicable operation in this area, at 75 ℃, toasts 12 hours.After cooling, Scotchpak1022 peel ply is placed on to obtained sticking, is placed on hypothallus to protect this hypothallus, finally obtain patch.
The thickness of this patch is 1.2mm, and bonding adhesive attraction is 0.62N.
Embodiment 5 nano-emulsion of the present invention promotes the Transdermal absorption of active constituents of medicine
Use rectilinear Franz diffusion cell (Hanson Research, the U.S.) to carry out vitro skin diffusion research.Described diffusion cell, with the reception tank of 7mL, has 3.8cm
2diffusion area.The porcine skin sample that about 1.1mm is thick is cut into 4.5-5cm
2sheet, be clipped between two compartments of diffusion cell, wherein stratum corneum side is to donor compartment, corium is towards receptor compartment.Receive and formed by normal saline solution (0.9%NaCl).In donor compartment, fill testing sample and corresponding control sample: the transdermal patch that testing sample adopts the present invention of embodiment 4.2 preparations above to comprise the nano-emulsion that contains Chinese herbal medicine aqueous and oiliness extract, its surface area is controlled in about 4cm
2; Control sample adopts above the prepared Chinese herbal medicine paste that contains Chinese herbal medicine aqueous and oiliness extract in embodiment 1.By external water circulation device, whole diffusion cell is maintained to 37 ℃ with the body temperature of simulation organism, and maintain the stirring of 600rpm.When 24h, to collect from the sample that receives compartment and porcine skin sample, the content that receives solution (reception phase) and porcine skin internal labeling thing by detection is analyzed to be measured and through performance control sample.Above-mentioned experiment in triplicate.
The representative active component of Flos Carthami, Radix Dipsaci, Radix Notoginseng and Radix Et Rhizoma Rhei (being chemical labeling) is listed in the table below:
Adopt HPLC-ESI-MS to detect label and the control sample seeing through.Use Agilent1290Infinity LC system (purchased from Aglient Technologies), its configuration vacuum degassing machine, binary liquid phase pump, automatic sampler, the triple level Four bars of Aglient6410 (Triple Quad) LC/MS, and be connected with Agilent MassHunter terminal (Workstation) software.Use Acquity UPLC HSS T31.8 μ m (2.1mm * 150mm) post, and make it remain on the temperature of 40 ℃.
The gradient that employing is comprised of water (A) and acetonitrile (B) flows to be separated and detects 9 chemical markers in sample.Gradient condition is 0-3min, 10-27%B; 3-5min, 27-33%B; 5-12min, 33-33%B; 12-13min, 33-80%B; 13-16min, 80-90%B; 16-20min, 90-90%B.Flow speed control is at 0.5mL/min, and volume injected is 20uL.
The triple level Four bar of the Aglient6410 LC/MS that MS analysis and utilization disposes ESI source carries out, it is monitored under negative ion mode and multiple reaction monitoring pattern, for the object ion of Hydroxy Carthamus yellow (HYA) application m/z611.2-> 325.0; For ginsenoside Rg1 (Rg1) application m/z799.5-> 637.4; For asperosaponin VI (ASP6) application m/z927.5-> 603.3; For ginsenoside Rb1 (Rb1) application m/z1107.6-> 119.0; For kaempferol (Kae) application m/z285.0-> 117.0; For emodin (Emo) application m/z269.0-> 241.0; For chrysophanic acid (Rhe) application m/z283.0-> 239.0; For oleanolic acid (OA) application m/z455.3-> 407.4.
testing result
Following table 1 has shown that testing sample (transdermal patch of nano-emulsion) and control sample (Chinese herbal medicine paste) see through skin diffusion to the percentage by weight of the label of reception tank
Table 1. diffuses to the percentage by weight of the label of reception tank
Following table 2 has shown that testing sample (transdermal patch of nano-emulsion) and control sample (Chinese herbal medicine paste) diffuse in skin or the percentage by weight of the label of dermis of skin side
Table 2. label diffuses to the percentage by weight of skin
According to the result of table 1 and table 2, draw Fig. 4 a and Fig. 4 b respectively; Wherein abscissa represents label, vertical coordinate represents to utilize the transdermal test data of Chinese herbal medicine paste as radix, the increase rate of label transdermal diffusion in the Chinese herbal medicine paste calculate obtaining and nano-emulsion/patch, thereby the diffusion of label in Chinese herbal medicine paste and nano-emulsion/patch relatively easily.
From Fig. 4 a, in above-mentioned permeability test, oleanolic acid (OA) is because its low aqueous solubility does not have to detect in receiving solution.For other seven kinds of labellings, the diffusion of the Hydroxy Carthamus yellow in nanometer patch (HYA) be enhanced 8 times, 14 times of ginsenoside Rg1s, 472 times of ginsenoside Rb1s, asperosaponin VI32 doubly, 11 times of 203 times of emodins, 21 times of chrysophanic acids and kaempferols.Adopt independent Flos Carthami, Radix Dipsaci, Radix Notoginseng and Radix Et Rhizoma Rhei extract to carry out permeability, obtain similar result (data are not shown).
By mensuration, be retained in the markd diffusion on porcine skin, from Fig. 4 b, the diffusion of the HYA in nanometer patch be enhanced 41 times, 21 times of Rg1,17 times of Rb1,33 times of ASP6, Emo5 doubly, Rhe11 doubly, OA47 doubly and Kae12 doubly.Adopt independent Flos Carthami, Radix Dipsaci, Radix Notoginseng and Radix Et Rhizoma Rhei extract to carry out permeability, obtain similar result (data are not shown).Visible, to compare with Chinese herbal medicine paste, nano-emulsion of the present invention and the patch that comprises it have extensively improved the transdermal diffusion of active constituents of medicine and have absorbed, and for some active component, this raising is remarkable especially.
Further experiment shows, when the prepared patch of embodiment 4.2 is applied to skin, promote Flos Carthami, Radix Dipsaci, Radix Notoginseng and the aqueous solvent extraction thing of Radix Et Rhizoma Rhei and the percutaneous of oil-based solvent extract to absorb, strengthened the therapeutic effect (data are not shown) for fracture.
As previously mentioned, although the present invention has described one or more embodiment herein, those of ordinary skills will appreciate that, can modify and not deviate from spirit of the present invention and essence the present invention.
Claims (17)
1. a herbal composite, the aqueous solvent extraction thing that it comprises Flos Carthami, Radix Dipsaci, Radix Notoginseng and Radix Et Rhizoma Rhei and oil-based solvent extract and optionally, pharmaceutically acceptable carrier, and wherein said herbal composite does not comprise the extract from Fructus Gardeniae and Ramulus Sambuci Williamsii.
2. according to the compositions of claim 1, it is comprised of following: the aqueous solvent extraction thing of Flos Carthami, Radix Dipsaci, Radix Notoginseng and Radix Et Rhizoma Rhei and oil-based solvent extract and optionally, pharmaceutically acceptable carrier.
3. according to the compositions of claim 1 or 2, wherein said aqueous solvent is that water and/or described oil-based solvent are ethanol, preferably 95% ethanol water.
4. according to the compositions of aforementioned claim any one, wherein the weight ratio of Flos Carthami, Radix Dipsaci, Radix Notoginseng and Radix Et Rhizoma Rhei is (0.8-1.2): (1-1.5): (0.8-1.2): (1-2), and preferably 1: 1: 1: 1 to 1: 1.5: 1: 2 scope; Preferably, the weight ratio of described aqueous solvent extraction thing and described oil-based solvent extract is 10: 1 to 7: 1, more preferably, the weight ratio of described aqueous solvent extraction thing and described oil-based solvent extract is counted 30%: 3% to 42%: 6% to account for the percentage by weight of described compositions, preferably 35%: 4.3% to 39.8%: 5%.
5. according to the compositions of aforementioned claim any one, it is prepared by following steps:
A) prepare Flos Carthami, Radix Dipsaci, Radix Notoginseng and Radix Et Rhizoma Rhei, optionally by its stripping and slicing;
B) step raw material a) is heated to the sufficiently long time under immersion and/or backflow jointly or respectively in aqueous solvent, to obtain aqueous solvent extraction thing, optionally gained aqueous solvent extraction thing is selected to filtration, concentrated and cryodesiccated processing;
C) by step raw material or step b a)) the extraction residue of gained soaks and/or the sufficiently long time of reflux in oil-based solvent, to obtain oil-based solvent extract, optionally gained oil-based solvent extract is selected to filtration, concentrated and cryodesiccated processing; With
D) the described aqueous solvent extraction thing and the described oil-based solvent extract that obtain are formulated as to compositions, wherein optionally by the described aqueous solvent extraction thing obtaining and described oil-based solvent extract and the combination of at least one pharmaceutically acceptable carrier.
6. according to the compositions of claim 1-4 any one, it is prepared by following steps:
A) prepare Flos Carthami, Radix Dipsaci, Radix Notoginseng and Radix Et Rhizoma Rhei, optionally by its stripping and slicing;
B) step raw material a) is heated to the sufficiently long time under immersion and/or backflow jointly or respectively in oil-based solvent, to obtain oil-based solvent extract, optionally gained oil-based solvent extract is selected to filtration, concentrated and cryodesiccated processing;
C) by step raw material or step b a)) the extraction residue of gained soaks and/or the sufficiently long time of reflux in aqueous solvent, to obtain aqueous solvent extraction thing, optionally gained aqueous solvent extraction thing is selected to filtration, concentrated and cryodesiccated processing; With
D) combination obtains described aqueous solvent extraction thing and described oil-based solvent extract, wherein optionally by the described aqueous solvent extraction thing obtaining and described oil-based solvent extract and the combination of at least one pharmaceutically acceptable carrier.
7. according to the compositions of aforementioned claim any one, it is the form of paste, ointment, nano-emulsion or percutaneous patch.
8. an oil-in-water or water-in-oil type nanoemulsion agent, it comprises water, organic facies and surfactant, the oil-based solvent extract that the aqueous solvent extraction thing that wherein said water comprises one or more Chinese herbal medicine materials and/or described organic facies comprise described one or more Chinese herbal medicine materials.
9. nano-emulsion according to claim 8, wherein said water or organic facies have 10-200nm, the preferred drop of 10-50nm; And/or described water accounts for the 60-90% of described nano-emulsion, preferred 75-85% by weight; And/or described organic facies accounts for the 2-20% of described nano-emulsion, preferred 2-8% by weight; And/or described surfactant accounts for the 4-40% of described nano-emulsion, preferred 10-22% by weight.
10. nano-emulsion according to claim 8 or claim 9, the aqueous phase solvent that wherein forms described water is water; And/or the oil phase solvent that wherein forms described oil phase is one or more combination of Sefsol-218, Capryol-90, triglyceride, Myritol-318, limonene, liquid paraffin, ethanol or its; And/or wherein said surfactant is to have scope at the non-ionic surface active agent of 8 to 18 HLB value, for example Labrasol, castor oil hydrogenated EL (Cremophor EL), polysorbas20, polysorbate60 and Tween 80, be preferably castor oil hydrogenated EL and polysorbas20.
Nano-emulsion described in 11. according to Claim 8-10 any one, wherein said aqueous solvent extraction thing and described oil-based solvent extract are respectively aqueous solvent extraction thing and the oil-based solvent extract limiting according to claim 1-6 any one.
Prepare oil-in-water described in claim 8-11 any one or the method for water-in-oil type nanoemulsion agent for 12. 1 kinds, it comprises the following steps:
1) utilize aqueous solvent extraction Chinese herbal medicine material to obtain aqueous solvent extraction thing, described aqueous solvent is preferably water;
2) utilize oil-based solvent to extract Chinese herbal medicine material to obtain oil-based solvent extract, described oil-based solvent is preferably alcohol, more preferably 95% ethanol;
3) by step 1) the aqueous solvent extraction thing that obtains is dissolved in aqueous phase solvent, to form the water that comprises described aqueous solvent extraction thing, and by step 2) the oil-based solvent extract that obtains is dissolved in oil phase solvent, to form the organic facies that comprises described oil-based solvent extract;
4) described water and described organic facies are mixed with surfactant, form the front mixture of homogenize;
5), with 12,000-30, the speed of 000rpm, by mixture homogenization 10-30 minute before described homogenize, forms described nano-emulsion.
13. 1 kinds of percutaneous patches, it comprises lining, hypothallus and peel ply, described hypothallus comprises oil-in-water type or water-in-oil type nanoemulsion agent and the binding agent described in claim 8-11 any one, and described nano-emulsion preferably accounts for the 1-25% of described hypothallus by weight; Wherein said binding agent is preferably selected from the group being comprised of Carbopol, carbopol U20, carbopol HV-805EG, sodium carboxymethyl cellulose, gelatin, polyacrylic acid sodium salt, polyacrylic acid sodium salt NP700, polyvinylpyrrolidone 10 and PVP K90 and combination thereof, and described binding agent is the combination of polyacrylic acid sodium salt, polyacrylic acid sodium salt NP700, carbopol HV-805EG and polyvinylpyrrolidone more preferably; And described binding agent preferably accounts for the 10-15% of described hypothallus by weight.
14. percutaneous patches according to claim 13, wherein said hypothallus further comprises one or more compositions of the group that is selected from cross-linking agent, wetting agent, timbering material or filler and pH controlling agent composition.
15. percutaneous patches according to claim 14, nano-emulsion, the binding agent of 10-15%, the wetting agent of the cross-linking agent of 0.01-0.8%, 15-25%, the timbering material of 1-10% or the filler that wherein said hypothallus comprises 1-25% with the weighing scale of substrate and the pH controlling agent that pH is controlled to 5-6.
The purposes of percutaneous patch described in compositions described in 16. claim 1-7 any one, the nano-emulsion described in claim 8-11 any one or claim 13-15 any one in the medicine for the preparation for the treatment of traumatic damage.
17. according to the purposes of claim 16, and wherein said traumatic damage is tissue injury or bone injury, and traumatic musculoskeletal injuries for example, such as soft tissue injury or fracture.
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|---|---|---|---|
| US14/290,962 US20140358094A1 (en) | 2013-05-29 | 2014-05-29 | Composition, the nano-emulsion and transdermal patch, methods of preparation and use thereof for treating traumatic injuries |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US201361855966P | 2013-05-29 | 2013-05-29 | |
| US61/855,966 | 2013-05-29 |
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| CN201410232730.4A Pending CN104208173A (en) | 2013-05-29 | 2014-05-29 | Chinese herbal medicine composition, nanometer emulsion and transdermal patch, and preparation thereof and purpose for traumatic injury therapy |
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| HK (1) | HK1199707A1 (en) |
Cited By (5)
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| CN105288016A (en) * | 2015-12-03 | 2016-02-03 | 广东省第二中医院 | Externally-applied traditional Chinese medicine emulsion for prevention and treatment of knee osteoarthritis and preparation method thereof |
| CN105395767A (en) * | 2015-12-02 | 2016-03-16 | 赵剑波 | Plaster for department of orthopaedics and preparation method of plaster |
| CN107982466A (en) * | 2017-12-27 | 2018-05-04 | 高永腾 | A kind of Chinese medicine composition for treating fracture and preparation method thereof |
| CN110161135A (en) * | 2018-02-13 | 2019-08-23 | 国药集团同济堂（贵州）制药有限公司 | The preparation method and its detection method of teasel root standard decoction |
| CN117482099A (en) * | 2024-01-03 | 2024-02-02 | 中日友好医院(中日友好临床医学研究所) | Application of akebia stem saponin D in preparation of medicines for resisting skeletal muscle atrophy and myopathy |
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| CN102579634A (en) * | 2012-02-23 | 2012-07-18 | 熊桂华 | Externally applied traditional Chinese medicine composition for treating knee osteoarthritis (OA) |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105395767A (en) * | 2015-12-02 | 2016-03-16 | 赵剑波 | Plaster for department of orthopaedics and preparation method of plaster |
| CN105288016A (en) * | 2015-12-03 | 2016-02-03 | 广东省第二中医院 | Externally-applied traditional Chinese medicine emulsion for prevention and treatment of knee osteoarthritis and preparation method thereof |
| CN107982466A (en) * | 2017-12-27 | 2018-05-04 | 高永腾 | A kind of Chinese medicine composition for treating fracture and preparation method thereof |
| CN110161135A (en) * | 2018-02-13 | 2019-08-23 | 国药集团同济堂（贵州）制药有限公司 | The preparation method and its detection method of teasel root standard decoction |
| CN110161135B (en) * | 2018-02-13 | 2022-03-11 | 国药集团同济堂（贵州）制药有限公司 | Preparation method and detection method of teasel root standard decoction |
| CN117482099A (en) * | 2024-01-03 | 2024-02-02 | 中日友好医院(中日友好临床医学研究所) | Application of akebia stem saponin D in preparation of medicines for resisting skeletal muscle atrophy and myopathy |
| CN117482099B (en) * | 2024-01-03 | 2024-04-09 | 中日友好医院(中日友好临床医学研究所) | Application of akebia stem saponin D in preparation of medicines for resisting skeletal muscle atrophy and myopathy |
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| HK1199707A1 (en) | 2015-07-17 |
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