CN113209007A - Asiaticoside preparation for external use and its preparation method - Google Patents

Asiaticoside preparation for external use and its preparation method Download PDF

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CN113209007A
CN113209007A CN202110532616.3A CN202110532616A CN113209007A CN 113209007 A CN113209007 A CN 113209007A CN 202110532616 A CN202110532616 A CN 202110532616A CN 113209007 A CN113209007 A CN 113209007A
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asiaticoside
external preparation
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sodium alginate
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范敏华
朱逸凡
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Zhejiang Poly Pharmaceutical Co ltd
Hainan Poly Pharmaceutical Co ltd
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Abstract

The invention belongs to the field of pharmaceutical preparations, and particularly relates to an external preparation for enhancing skin hydration degree and promoting asiaticoside transdermal absorption and a preparation method thereof. The asiaticoside external preparation comprises asiaticoside, gel matrix and other medicinal adjuvants. The asiaticoside external preparation of the invention takes potassium lactate and water-soluble azone as moisturizing penetrant, softens the cuticle of skin and enhances the transdermal effect of the asiaticoside, solves the problem that the asiaticoside is not easy to be absorbed transdermally, and is beneficial to the release of the asiaticoside.

Description

Asiaticoside preparation for external use and its preparation method
Technical Field
The invention belongs to the field of pharmaceutical preparations, and particularly relates to an external preparation for enhancing skin hydration degree and promoting asiaticoside transdermal absorption and a preparation method thereof.
Background
There are two forms of wound healing: one is complete recovery, i.e., repair by cells of the same structure as the original damaged tissue, such as scarless healing of an early wound of a fetus, or healing of a superficial wound; in addition to this, almost all wounds end up healing cicatricial at the same time as epithelialization. This scar is a product of the normal tissue repair process and is therefore called a "normal scar" (normal scar). The other is that when the wound repair process is abnormal, extracellular matrix components mainly containing collagen are deposited in a large quantity, dermal tissue is excessively proliferated, and a 'pathological scar' (abnormal scar) is generated, namely a 'hypertrophic scar' (HS) or a 'keloid' (K), and is a pathological state that extracellular matrix (ECM) components including types I and III collagen are excessively deposited in tissues and are difficultly absorbed or remolded by the body.
Asiaticoside (Asiaticoside) is a triterpenoid extracted from Centella asiatica (L.) Urban of Umbelliferae. Studies show that asiaticoside can reduce the expression of TGF-beta 1mRNA for promoting scar hyperplasia and increase the expression of TGF-beta 3mRNA for inhibiting scar hyperplasia, so that the expression of metal matrix protease inhibitor (TIMPS) is obviously reduced, and the degradation of type I collagen is promoted; can also inhibit TGF-beta pathological action, prevent fibroblast proliferation, weaken activity of synthesized and secreted protein and relieve scar hyperplasia by increasing expression of Smad7 of inhibitory R-Smads transduction signals. A large number of clinical practices have proved that the external asiaticoside preparation has definite therapeutic effects on promoting wound healing and inhibiting the formation of skin scar on human skin.
Asiaticoside is a pentacyclic triterpenoid saponin, has the characteristics of hydrophobic aglycone structure and hydrophilic sugar chain structure in the molecular structure, weak acidity, extremely slight dissolution in aqueous solution, poor lipophilicity and the like, is not beneficial to diffusion through stratum corneum and has low bioavailability, so the transdermal application of the asiaticoside is limited. How to make asiaticoside pass through the stratum corneum of skin better, act on the dermis and improve bioavailability is always a hot point of research.
There are two main aspects in the current research direction of external asiaticoside preparations. Firstly, the dosage form of the asiaticoside external preparation is changed, for example, the asiaticoside is prepared into microemulsion and liposome, so that the capability of passing through a stratum corneum barrier is improved to improve the bioavailability; or prepared into gel, and the purpose of increasing the bioavailability is achieved by utilizing the functions of strong adhesiveness and long retention time of the gel on the skin to increase the release and absorption time. On the other hand, the variety selection and the addition amount of a skin penetration enhancer or the composite use of several penetration enhancers are studied to promote the absorption of asiaticoside, for example, Wangshanping et al study the promotion effect of three penetration enhancers of azone, menthol and borneol on the transdermal penetration of asiaticoside, and the study shows that the penetration-promoting effect of menthol and borneol with the same concentration is obvious and is obviously better than that of azone with the same concentration. The above mentioned solutions all achieve certain results, but each has disadvantages, as shown in the following table:
Figure BDA0003068498450000021
therefore, how to avoid the occurrence of the disadvantages as much as possible on the premise of keeping the advantages is of great significance to the application of the asiaticoside.
The gel is a semisolid or liquid preparation with certain viscosity prepared by mixing the medicine and a proper gel matrix, has uniform and fine texture, can form a layer of film on the skin, has strong adhesiveness, long retention time and no irritation to the skin and mucous membrane, can generate the sustained-release or controlled-release medicine release effect, has good skin coating performance, convenient use and simple preparation process, and has a plurality of advantages when being used as a skin external preparation.
Sodium alginate is a by-product after extraction of iodine and mannitol from brown algae such as kelp or gulfweed. The sodium alginate contains a large amount of-COO-, shows polyanion behavior in aqueous solution, has certain adhesiveness, can be used as a drug carrier, has the effect of promoting the growth of neurovascular at wounds, and is very favorable for the healing of the wounds. When cations such as Ca2+, Sr2+ and the like exist in the sodium alginate solution, the sodium alginate solution is crosslinked to form hydrogel. The sodium alginate hydrogel used as a preparation material of the dressing for wound healing has good biocompatibility and biodegradability, does not stimulate and generate immune response to human bodies, has strong moisture retention property, can keep the surface of a wound moist, relieve pain, avoid dry scab formation and greatly shorten the time for wound healing, and is beneficial to the hydration of the surface of the skin, so that the absorption of asiaticoside is promoted, the healing is promoted and the formation of scars is inhibited. However, sodium alginate hydrogel has poor strength, is easy to break, and has poor skin coating and adhesion properties, so that the application is less. The polyvinyl alcohol is an extremely safe high-molecular organic matter, has no toxicity or side effect on a human body, has good biocompatibility, can be used for preparing a membrane material, can well improve the strength of sodium alginate under a certain use proportion, enhances the skin coating and adhesion performance of sodium alginate hydrogel, plays a role in balancing the pH value of the sodium alginate gel on the other hand, and is beneficial to the release of asiaticoside.
Traditional researches prove that the skin hydration degree has positive correlation to the promotion of drug absorption, because stratum corneum cells can absorb a certain amount of water, and self-expansion reduces the structural compactness, thereby promoting drug permeation, and particularly the increase influence of the bound water content of the stratum corneum cells is more obvious, and research data shows that when the stratum corneum content reaches 50%, the permeation and absorption capacity of the drug is enhanced by 5-10 times. Therefore, how to enhance the skin hydration degree, especially the content of the water bound in the stratum corneum cells, is also of great significance for promoting the transdermal absorption of asiaticoside and enhancing the treatment effect. The components with the functions of locking water and keeping moisture in a certain proportion can be used
In order to promote the absorption of the drug, a penetration enhancer is usually used in skin external preparations, and azone can make the skin stratum corneum interact with lipid, reduce the phase transition temperature of the drug to the lipid in the stratum corneum space, increase the fluidity, and reduce the diffusion resistance of the drug or active additives in the stratum corneum, so that the skin external preparations have strong penetration enhancing effect, are commonly used penetration enhancers, and the penetration enhancing effect is increased along with the increase of the use concentration. However, in the healing process of the wound tissue, due to the deposition of collagen, scars are formed, the collagen fibers are thinned, the stratum corneum is absent or is forming, the growth environment in the wound or the scar tissue is changed due to the use of the permeation enhancer with higher concentration, the wound is difficult to heal, or the collagen fibers are deposited disorderly, so that pathological scars are generated. Therefore, on the premise of promoting the transdermal absorption of asiaticoside, how to minimize the use of a penetration enhancer is important for promoting wound healing and inhibiting the generation of scars.
Disclosure of Invention
The invention aims to provide an asiaticoside external preparation which is prepared by combining a gel matrix and a moisturizing penetration enhancer, enhances the hydration degree of skin and promotes the transdermal absorption of asiaticoside.
In order to achieve the purpose of the invention, the invention adopts the following technical scheme:
an asiaticoside external preparation comprises asiaticoside, gel matrix and other medicinal adjuvants.
The asiaticoside external preparation is characterized in that the matrix material is one or the combination of more of carbomer, sodium alginate and polyvinyl alcohol.
Preferably, the asiaticoside external preparation and other pharmaceutical excipients comprise a moisturizing penetration enhancer.
Preferably, the matrix material is sodium alginate and polyvinyl alcohol.
Preferably, the weight percentage of the sodium alginate to the polyvinyl alcohol is 10:0.5-10: 3.
Preferably, the moisture-retention penetration enhancer is water-soluble azone and potassium lactate.
Preferably, the weight percentage of the potassium lactate to the water-soluble azone is 2:1-20: 1.
Preferably, the weight percentage of the potassium lactate and the water-soluble azone is 12: 1.
preferably, the other pharmaceutical excipients also include solvents and PH regulators.
The asiaticoside external preparation of the invention takes potassium lactate and water-soluble azone as moisturizing penetrant, softens the cuticle of skin and enhances the transdermal effect of the asiaticoside, solves the problem that the asiaticoside is not easy to be absorbed transdermally, simultaneously has the water locking function, particularly the water binding of the cells of the horny layer, enhances the penetrating and absorbing capacity of the medicine, has the characteristics of good moisturizing effect and high skin coating performance, adopts sodium alginate and polyvinyl alcohol as gel matrixes to obtain gel with good biocompatibility and biodegradability, has strong moisture retention, is beneficial to the hydration of the skin surface so as to promote the absorption of asiaticoside, meanwhile, the polyvinyl alcohol is adopted to improve the strength of the sodium alginate, enhance the skin coating and adhesion performance of the sodium alginate hydrogel, and play a role in balancing the pH value of the sodium alginate gel, thereby being beneficial to the release of the asiaticoside.
Drawings
FIG. 1: a asiaticoside solution standard curve chart;
FIG. 2: a graph comparing the transdermal steady state release rates;
FIG. 3: example 1 asiaticoside gel intradermal retention assay liquid phase diagram;
FIG. 4: a liquid phase diagram of a commercially available asiaticoside cream ointment retention test in the skin is shown.
Detailed Description
Example 1: 1000g of asiaticoside gel was prepared using the following materials:
25g of asiaticoside, 50g of glycerol, 40g of propylene glycol, 15g of sodium alginate, 1.5g of polyvinyl alcohol, 12g of potassium lactate, 1g of water-soluble azone, 4g of triethanolamine and 851.5g of water.
The preparation method comprises the following steps:
1. mixing asiaticoside, glycerol and propylene glycol according to the prescription amount by vortex, and heating to 80 ℃ to obtain an asiaticoside solution;
2. dissolving potassium lactate 12g and water soluble azone 1g in water to obtain potassium lactate azone solution, adding the asiaticoside solution into the potassium lactate azone solution, and stirring.
3. Taking the rest water, dispersing sodium alginate and PVA in the water to completely swell, adding the solution obtained in the step 2 into the water, and uniformly stirring at a low speed.
4. And (3) adding triethanolamine into the solution obtained in the step (3), and uniformly stirring at a low speed.
The asiaticoside sodium alginate gel obtained in the embodiment has a simple preparation process, adopts sodium alginate and polyvinyl alcohol as gel matrixes, and is added with potassium lactate and azone for compound use, so that the addition of a penetration enhancer azone is reduced by 0.1%, and the asiaticoside sodium alginate gel has good biocompatibility, and has the characteristics of no adverse reaction and high safety when used in early stages of healing of fresh wounds and wounds. The asiaticoside sodium alginate gel obtained by the embodiment has strong water-locking and moisture-keeping properties, improves the skin hydration degree, greatly improves the transdermal absorption rate of the asiaticoside, and improves the bioavailability.
Comparative experiment
The test sample was a sodium alginate asiaticoside gel prepared according to example 1.
The control sample adopts a commercially available asiaticoside cream ointment, the manufacturer is Shanghai modern pharmaceutical Co., Ltd, the content is 2.5%, and the specification is as follows: 30 g/piece.
The analysis method comprises the following steps: HPLC method
Octadecylsilane chemically bonded silica is used as a filling agent; acetonitrile-2 mmol/L beta-cyclodextrin solution (24:76) is taken as a mobile phase; the sample amount is 10 mul; the detection wavelength was 205 nm. The theoretical plate number is not less than 5000 calculated according to asiaticoside peak.
Taking appropriate amount of asiaticoside reference substance, precisely weighing, and adding methanol to obtain solution containing asiaticoside 0.2mg, asiaticoside 0.4mg, asiaticoside 0.8mg, asiaticoside 1.0mg and asiaticoside 1.2mg in each 1ml, with linear range of 0.1-5 ug. The standard curve equation is prepared as A ═ 2225.9m +4.8933, R20.9997 (where m is the amount of asiaticoside taken in and a is the peak area). See fig. 1.
The Content (CX) in the permeated solution was calculated from the peak area of asiaticoside in mg/ml.
Test method 1 transdermal diffusion test
The pretreated mouse abdomen depilatory skin is fixed between a dosing pool and a receiving pool of a Frangz diffusion pool, and 1g of asiaticoside cream ointment serving as a control sample and 1g of sodium alginate gel serving as a test sample are respectively and uniformly coated on the mouse skin. The receiving solution was a 20% ethanol physiological saline solution, and the dermal side was brought into complete contact with the receiving solution, and the contact surface was carefully cleaned of air bubbles. The volume of the receiving solution is 6.5ml, and the effective diffusion area is 2.87cm2The test temperature was 37. + -. 1 ℃ and the stirring speed was 800 rpm. Sampling 1ml at 2, 4, 6, 8 and 24h (simultaneously supplementing the same-temperature equal-quantity receiving solution), filtering with 0.22um filter membrane, and determining asiaticoside content according to the above analysis method. Each sample was tested in parallel 3 times, and the cumulative permeation amount per unit area Q was calculated according to the following formula (1)1/ug·cm-2And (2) calculating the percutaneous steady-state permeation rate Js/ug cm-2/h-1
Formula (1): q1(content (CX) × 6.5 × 10)3)/2.87
Formula (2): js ═ Q1T and t are respectively 2, 4, 6, 8 and 24 h.
Plotting t against Q1, the results are shown in FIG. 2:
the steady-state release rate of the asiaticoside gel prepared in the specific example 1 is obviously superior to that of the asiaticoside cream ointment of a commercial control sample; the 24-hour total penetration of the asiaticoside gel was 1.75 times the 24-hour total penetration of the commercial control asiaticoside cream ointment (76.41%, 43.73%, respectively). The excellent bioavailability of the asiaticoside gel is reflected.
The test method 2: determination of Release Capacity
After the transdermal test is finished, the in vitro skin is immediately taken down, a cotton swab is used for removing a tested sample, the skin is washed clean by PBS, filter paper absorbs water, the skin is cut into pieces and then placed into a 5ml centrifuge tube, 1ml of methanol is added for soaking overnight, the mixture is filtered through a 0.22um filter membrane, and the content of asiaticoside is measured according to the analysis method. The intradermal retention (Q) of the sample was calculated2) Total permeability Q calculated according to the following formula (3) in combination with test method 1)tThe release levels of the different samples were compared.
Formula (3) Qt＝Q1+Q2
The results are shown in FIGS. 3 and 4:
the asiaticoside cream ointment retains an asiaticoside integral peak area 451.24 in the skin, and the retention amount is calculated to be 20.05ug according to a standard curve equation; the total release amount is 10952.55ug calculated according to the above formula 3
The asiaticoside gel retained the integrated asiaticoside peak area in the skin of 345.96, and the retention was calculated to be 15.32ug according to the standard curve equation. The total release was calculated to be 194117.82ug according to equation 3 above.
The relationship between the 24-hour release amount and the retention amount is shown in Table 1 below.
Sample (I) Test sample amount (g) Released amount (ug) Release ratio Retention of intradermal Retention ratio
Ointment
1 10952.55 43.81％ 20.05 0.08％
Gel
1 19117.82 76.47％ 15.32 0.06％
The test result shows that the asiaticoside gel has obvious release performance. The asiaticoside gel is slightly better than commercially available asiatic pennywort ointment in terms of permeability.

Claims (9)

1. An asiaticoside external preparation comprises asiaticoside, gel matrix and other medicinal adjuvants.
2. The external asiaticoside preparation according to claim 1, wherein said matrix material is one or more selected from carbomer, sodium alginate and polyvinyl alcohol.
3. The asiaticoside external preparation according to claim 2, wherein the other pharmaceutical excipients comprise a moisture-retaining penetration enhancer.
4. The asiaticoside external preparation according to claim 3, wherein said base material is sodium alginate or polyvinyl alcohol.
5. The asiaticoside external preparation according to claim 4, wherein the weight percentage of sodium alginate to polyvinyl alcohol is 10:0.5-10: 3.
6. An asiaticoside external preparation according to claim 3, wherein said moisture-retaining penetration enhancer is water-soluble azone and potassium lactate.
7. The asiaticoside external preparation according to claim 6, wherein the weight percentage of said potassium lactate and said water-soluble azone is 2:1 to 20: 1.
8. Asiaticoside external preparation according to claim 6, wherein said potassium lactate and water-soluble azone are present in a weight ratio of 12: 1.
9. the asiaticoside external preparation according to claim 1, wherein the other pharmaceutical excipients further comprise a solvent and a PH regulator.
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