CN102846546B - Percutaneous-absorption-promoting propranolol composite phospholipid transfersome, and prepartion method and application thereof - Google Patents
Percutaneous-absorption-promoting propranolol composite phospholipid transfersome, and prepartion method and application thereof Download PDFInfo
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Abstract
The invention provides a composite phospholipid transfersome which promotes propranolol percutaneous absorption, and a preparation method thereof. According to the invention, two phospholipid materials with different phase-change temperatures, which are dipalmitoyl phosphatidyl choline and soybean lecithin, are adopted as a composite phospholipid material. Compared with a transfersome with a single phospholipid material in prior art, the propranolol composite phospholipid transfersome prepared with the phospholipid material provided by the invention has substantially improved encapsulation efficiency, reduced leakage, improved stability in rat plasma, and substantially improved bioavailability after percutaneous administration. The propranolol composite phospholipid transfersome provided by the invention is especially suitable to be used for treating infantile hemangioma. With the transfersome, propranolol percutaneous administration can be realized, and propranolol can directly act upon a hemangioma affected part. The treatment effect is improved, toxic and side effects are reduced, and children medication compliance can be improved. Also, the invention provides a preparation method of the propranolol composite phospholipid transfersome.
Description
Technical field
The present invention relates to technical field of medicine, be specifically related to a kind of composite phospholipid carrier that can promote the absorption of Propranolol percutaneous and preparation method thereof, the application of this kind of Propranolol composite phospholipid carrier in treatment infantile hemangioma is also provided simultaneously.
Background technology
Propranolol (Propranolol), as a kind of non-selective beta-blocker of classics, is now extensively confirmed to have the precise effects for the treatment of infant hemangioma (Infantile Hemangiomas, IH).At present, Propranolol is the oral administrations that adopt while treating for IH more, but because IH treatment cycle is long, this administering mode compliance for infant is poor, and may cause the untoward reaction widely such as bradycardia, hypotension, hypoglycemia, and it is serious that Propranolol oral administration has been proved first pass effect, and bioavailability is low.If therefore can and directly act on IH affected part by Propranolol transdermal administration, when guaranteeing curative effect, can effectively reduce or remit the generation of untoward reaction, and administration is convenient, good to patient's compliance.Yet the outermost horny layer of intact skin, especially epidermis, as the main barrier of most percutaneous drug absorptions, has greatly restricted transdermal drug delivery system (Transdermal Drug Delivery System, TDDS) application clinically.
Carrier is a kind of self aggregation vesicle growing up on the basis of liposome, its composition is generally in phospholipid bilayer, to mix edge activating agent, thereby give its special flexibility, can be under external pressure, crimp is only the duct of self particle diameter 1/10 through aperture, so carrier except having the feature that conventional liposome hydrophilic is strong, also has morphotropism and the transdermal capability of height, so become extremely potential novel transdermal administration carrier system.
The physicochemical properties that carrier need to be investigated mainly comprise: morphology, particle diameter and distribution thereof, Zeta potential, envelop rate, stability etc.Wherein, in view of carrier mainly strengthens the percutaneous abilities of medicine with the form of carrier, therefore improve carrier, the envelop rate of medicine is just seemed to particularly important; And the distinctive marks that morphotropism is different from conventional liposome as carrier can be predicted the percutaneous abilities of carrier effectively, is content very important in carrier property research.
The phospholipid material adopting in carrier preparation process is similar to conventional liposome, and conventional have soybean phospholipid (SPC), HSPC (HPC), egg PC (EPC), dipalmitoyl phosphatidyl choline (DPPC), a distearoyl phosphatidylcholine (DSPC) etc.In general, the phospholipid (as DSPC, HSPC) of high phase transition temperature (more than 37 ℃), because rigidity is higher, is unfavorable for the distortion after carrier administration, therefore less employing.Conventionally adopt the phospholipid of low transformation temperature (37 ℃ following) as film material, but because this kind of its phase transition temperature of medicine carrying carrier is lower than body temperature, after administration, under the effect of body temperature, phospholipid bilayer changes liquid crystalline phase into, drug leakage is more, is unfavorable for the Transdermal absorption of medicine carrying carrier.And if in carrier preparation, only adopt a kind of phospholipid material will inevitably cause this kind of situation to occur as film material, so if phase transition temperature that can flexible phospholipid is expected to realize high morphotropism and the high stability of carrier simultaneously.
2004, Kan P etc. invented the composite phospholipid liposome technology of preparing carrier with multiple phospholipid material, and its basic feature is to use the phospholipid of different phase transition temperatures as film material, as conventional soybean phospholipid (SPC, T
mbe about 0 ℃), egg phosphatide (EPC, T
mbe about-8 ℃), distearoyl phosphatidylcholine (DSPC, T
mbe about 55 ℃), hydrogenated soya phosphatide (HSPC, T
mbe about 50 ℃) etc.Under certain condition, in composite phospholipid, two kinds of phospholipid can merge and become cenotype, and its phase transition temperature can be by changing the ratio flexible of two kinds of phospholipid.
From current technology, the Propranolol of percutaneous dosing has the report of making the dosage forms such as patch, but infant hemangioma area is unfixing, and skin surface is also difficult for long-time stickup, adopts liquid or semisolid dosage form to be more suitable for undoubtedly the needs of clinical practice.But the direct administration polarity of Propranolol is large, Transdermal absorption amount is limited, and the carrier technology of applying single Lipid composition can promote the Transdermal absorption of Propranolol, but degree is limited.
Summary of the invention
Technical solution problem: in order to solve the deficiencies in the prior art, the invention provides a kind of composite phospholipid carrier that can promote that Propranolol percutaneous absorbs; Another object of the present invention is to provide a kind of method of preparing described Propranolol composite phospholipid carrier; The present invention simultaneously also provides the application of described Propranolol composite phospholipid carrier in treatment infant hemangioma.And composite phospholipid carrier provided by the present invention, the adjusting consisting of composite phospholipid due to phase transition temperature is set in 37 ℃ of left and right, therefore in transdermal drug delivery system, has maximum mechanism.
Technical scheme: for realizing above object, the invention provides a kind of Propranolol composite phospholipid carrier that can promote percutaneous drug absorption, it is characterized in that it is by two class phospholipid compositions: it is made that dipalmitoyl phosphatidyl choline and soybean phospholipid, edge activating agent and Propranolol are pressed row weight portion proportioning: 1 ~ 10 part of dipalmitoyl phosphatidyl choline (DPPC), 1 ~ 10 part of soybean phospholipid (SPC), 0.2 ~ 2 part of edge activating agent, the amount of Propranolol is 0.05 ~ 0.5 times of phospholipid total amount.
As a further improvement on the present invention, dipalmitoyl phosphatidyl choline equals 8:2 to 2:8 with the mole ratio of soybean phospholipid.
As of the present invention, further improve, dipalmitoyl phosphatidyl choline equals 6:4 with the mole ratio of soybean phospholipid.
As a kind of preferred scheme of the present invention, edge activating agent is selected from sodium cholate, NaTDC, tween and span.
As the scheme of present invention further optimization, edge activating agent is Arlacel-80.
Meanwhile, the present invention also provides a kind of method of preparing above-mentioned Propranolol composite phospholipid carrier, is divided into the blank composite phospholipid carrier of preparation and the large step of medicine carrying two, and its concrete operation step is as follows:
First step: prepare blank composite phospholipid carrier, comprising:
(1) take by weight 1 ~ 10 part of dipalmitoyl phosphatidyl choline, 1 ~ 10 part of soybean phospholipid, 0.2 ~ 2 part of edge activating agent, is dissolved in dehydrated alcohol, injects ammonium sulfate solution;
(2) by the method heating or reduce pressure, remove ethanol;
(3) supersound process or high pressure breast are even, reduce the particle diameter of composite phospholipid carrier;
(4) by dialysis, ultrafiltration or gel filtration chromatography method, remove the ammonium sulfate of not sealed by composite phospholipid carrier, form blank composite phospholipid carrier suspension.
Second largest step, medicine carrying, comprising:
(5) getting total amount is that the Propranolol of 0.05 ~ 0.5 times of phospholipid total amount adds temperature in above-mentioned blank composite phospholipid carrier suspension to incubate, and it is sealed by composite phospholipid carrier, makes Propranolol composite phospholipid carrier.
As a further improvement on the present invention, described in step (1) be dissolved in dehydrated alcohol time, with the speed of 5 ml/min, inject the ammonium sulfate solution of 0.1 ~ 0.3 mol/L of 10 ~ 20 times of volumes.
As present invention further optimization scheme, in step (4), adopt the phosphate buffer of pH7.4 to dialyse to remove the ammonium sulfate of not sealed by composite phospholipid carrier.
As another kind of preferred version of the present invention, it is 0 ~ 60 ℃ that the temperature described in step (5) is incubated temperature, and the temperature time of incubating is 5 ~ 60 min.
The present invention simultaneously also provides the application of described Propranolol composite phospholipid carrier in treatment infant hemangioma.
Beneficial effect
The present invention screens the phospholipid material of two kinds of different phase transition temperatures through lot of experiments: dipalmitoyl phosphatidyl choline and soybean phospholipid, through lot of experiments, show when the composite phospholipid material being formed by the DPPC of 1 ~ 10 part of molar fraction and the SPC of 1 ~ 10 part of molar fraction, when especially the mole ratio of DPPC and SPC equals 6:4, while being prepared into composite phospholipid carrier film, form cenotype, do not exist and be separated, there is single phase transition temperature.So just make the Propranolol composite phospholipid carrier being prepared into this composite phospholipid material, with in prior art, only adopt the carrier of single phospholipid material DPPC or SPC and compare, envelop rate is significantly improved, release in vitro presents slow release effect, stability in rat plasma improves, and after percutaneous dosing, under blood drug level and drug-time curve, area also significantly improves.Show can promote Propranolol percutaneous to absorb through the Propranolol carrier of composite phospholipid material parcel provided by the invention, bioavailability significantly improves, be particularly useful for treating the diseases such as infant hemangioma, can realize Propranolol transdermal administration and act directly on IH affected part, to realizing the collaborative performance of Propranolol part and whole body therapeutic effect, when improving curative effect, reducing toxic and side effects, improve the compliance of IH infant medication.
Accompanying drawing explanation
The HPLC figure of the blank carrier of Fig. 1 and Propranolol carrier
The impact of Fig. 2 preparation method on Propranolol carrier envelop rate, in figure, vertical coordinate EE represents entrapment efficiency
The impact of Fig. 3 edge activating agent on Propranolol carrier envelop rate, in figure, vertical coordinate EE represents entrapment efficiency
The morphology of the different Lipid composition Propranolol of Fig. 4 carrier
The impact of Fig. 5 Lipid composition on Propranolol carrier morphotropism, in figure, vertical coordinate is Δ DI=DI37-DI32, is used for describing the morphotropism situation of change that different Lipid composition Propranolol carriers cause because of temperature contrast
Fig. 6 does not add the L%-t curve (n=3, x ± s) of rat plasma, and in figure, vertical coordinate L% represents drug accumulation leakage percentage rate
Fig. 7 adds the L%-t curve (n=3, x ± s) of rat plasma, and in figure, vertical coordinate L% represents drug accumulation leakage percentage rate
Propranolol HPLC figure in Fig. 8 rat plasma
The blood drug level comparison of each test group of Fig. 9 (n=5, x ± s), in figure, the little figure blood drug level in the upper right corner is at 250 ngmL
-1when following, respectively organize the enlarged drawing of plasma concentration curve
The mensuration collection of illustrative plates of Figure 10 Propranolol composite phospholipid carrier phase transition temperature
The specific embodiment
Below in conjunction with specific embodiment, further illustrate the present invention, should understand these embodiment is only not used in and limits the scope of the invention for the present invention is described, after having read the present invention, those skilled in the art all fall within the application's claims limited range to the modification of the various equivalent form of values of the present invention.
The preparation of the blank phospholipid carrier of embodiment 1
Take 2 ~ 20 parts of phospholipid materials, its overall control is at 0.2 mmol, be dissolved in altogether appropriate dehydrated alcohol with 0.2 ~ 2 portion of edge activating agent, inject 0.2 mol/L ammonium sulfate solution of water bath with thermostatic control and magnetic agitation, by the method heating or reduce pressure, wave most ethanol, then take out and let cool, and be settled to 5 mL with ultra-pure water; 200 W Probe Ultrasonic Searching certain hours are to reduce the particle diameter of phospholipid carrier, low-speed centrifugal removes titanium valve and is placed in bag filter, with pH 7.4 phosphate buffers, dialyse to remove the ammonium sulfate that outer water is not sealed by phospholipid carrier again, obtain blank composite phospholipid carrier.
The preparation of embodiment 2 Propranolol phospholipid carriers
Under the basis of the blank phospholipid carrier of preparing at embodiment 1, given medicine/phospholipid=19.83%, gets a certain amount of Propranolol and blank phospholipid carrier in molar ratio, and water bath with thermostatic control temperature is incubated certain hour, Propranolol is sealed by phospholipid carrier, made Propranolol composite phospholipid carrier.
The mensuration of embodiment 3 Propranolol phospholipid carrier envelop rates
Concrete operation step divides three steps to carry out:
1. sample treatment: the Propranolol phospholipid carrier crude product sample that adopts the separation of Sephadex G-50 post to be prepared by embodiment 2, obtains pastille carrier and free drug, equal-volume rupture of membranes agent rupture of membranes after pastille carrier standardize solution, 12000 rmin
-1centrifugal rear standby.
2. the content of high-performance liquid chromatogram determination pastille carrier and free drug: the accurate upper high performance liquid chromatography Kromasil C18 post (250 mm * 4.6 mm, 5 μ m) of sample 1mL of drawing carries out eluting, and mobile phase is methanol-0.02 molL
-1kH
2pO
4(50:50), flow velocity 0.9 mLmin
-1, 35 ℃ of column temperatures, detect wavelength 289 nm, sampling volume 10 μ L.Under this chromatographic condition, can make pastille carrier and non-encapsulated free drug realize baseline separation, and instrument precision is good, the response rate is high, and carrier film material component is noiseless to the assay of Propranolol simultaneously, sees Fig. 1.
3. envelop rate calculates:
Envelop rate=carrier Chinese medicine content/(carrier Chinese medicine content+free drug content) * 100%
The selection of embodiment 4 Propranolol phospholipid carrier preparation methoies
Choose Arlacel-80 (Span-80) as edge activating agent, by four kinds of methods such as thin film aquation method, alcohol injection, ammonium sulphate gradient and reverse evaporations, prepare Propranolol carrier, parallel 3 parts of every group of sample, concrete steps are as follows:
(1) thin film aquation method: 0.2 mmol SPC and appropriate Span-80 are placed in the eggplant-shape bottle of 50 mL specifications, add 5 mL dehydrated alcohol, after fully dissolving, reduce pressure rotary evaporation to forming uniform class membrane of lipoprotein under 55 ℃ of water-baths, vacuum drying spends the night to remove the organic solvent of trace.To precision in gained class membrane of lipoprotein, add 5 mL containing the phosphate buffer (PBS) of 11.84 mg propranolol hydrochlorides (that is: medicine/phospholipid mol ratio is 0.2), and vibration is to abundant aquation under 55 ℃ of water-baths, gained pastille carrier suspension is through 200 W Probe Ultrasonic Searching 200 times.
(2) alcohol injection: 0.2 mmol SPC and appropriate Span-80 are dissolved in 5 mL dehydrated alcohol altogether, are injected into 5 mL containing in the PBS of 11.84 mg propranolol hydrochlorides, is stirred to and waves most ethanol in 55 ℃ of water-bath lower magnetic forces, through 200 W Probe Ultrasonic Searching 200 times and get final product.
(3) ammonium sulphate gradient: 0.2 mmol SPC and appropriate Span-80 are dissolved in 5 mL dehydrated alcohol altogether, being injected into 5 mL concentration is in the ammonium sulfate of 0.2 MolL-1, in 55 ℃ of water-bath lower magnetic forces, be stirred to and wave most ethanol, after 200 W Probe Ultrasonic Searching 200 times, low-speed centrifugal removes titanium valve and is placed in bag filter, with PBS dialysis (10 times of volumes, 4 times), to remove the ammonium sulfate of outer water, obtain blank carrier again.According to medicine/phospholipid mol ratio, be 0.2, get quantitative blank carrier and propranolol hydrochloride, 55 ℃ of water-bath vibration certain hours, obtain medicine carrying carrier.
(4) reverse evaporation: 0.2 mmol SPC and appropriate Span-80 are dissolved in 5 mL chloroform altogether, and fully mix containing the PBS of 11.84 mg propranolol hydrochlorides in 2.5 mL, 400W Probe Ultrasonic Searching makes to form stable w/o type Emulsion for a moment, under 55 ℃ of water-baths, reduce pressure rotary evaporation to forming semi-solid jelly, add appropriate PBS to vibrate under 55 ℃ of water-baths complete to aquation, gained pastille carrier suspension is through 200W Probe Ultrasonic Searching 200 times.
Preparing gained carrier, by method in embodiment 3, measure envelop rate, finally record envelop rate and the results are shown in Figure 2.Result shows, adopts initiatively the ammonium sulphate gradient of medicine carrying to prepare Propranolol and transmits physical ability and realize better medicine carrying effect.
The selection of embodiment 5 edge activating agents
By method in embodiment 2, prepare four groups of Propranolol carriers, wherein phospholipid is SPC, edge activating agent is respectively sodium cholate, NaTDC, Span-80 and tween 20, its consumption is consistent, preparing gained carrier, by method in embodiment 3, measure envelop rate, parallel 3 parts of every group of sample, finally records envelop rate and the results are shown in Figure 3.Result shows: the Propranolol carrier that the Span-80 of usining is prepared as edge activating agent is having significant advantage aspect raising entrapment efficiency.
The pharmaceutical properties evaluation of embodiment 6 Propranolol composite phospholipid carriers
By method in embodiment 2, under difference prescription condition, prepare seven groups of Propranolol phospholipid carriers, 3 parts of every group of parallel preparations, the condition of specifically writing out a prescription is as follows:
| Sequence number | Group name | Edge activating | Phospholipid material | |
| 1 | SL | Do not add | SPC | |
| 2 | ST | Span-80 | |
|
| 3 | NT-28 | Span-80 | DPPC:SPC=2:8 | |
| 4 | NT-46 | Span-80 | DPPC:SPC=4:6 | |
| 5 | NT-64 | Span-80 | DPPC:SPC=6:4 | |
| 6 | NT-82 | Span-80 | DPPC:SPC=8:2 | |
| 7 | DT | Span-80 | DPPC |
(1) envelop rate evaluation
The envelop rate of measuring not on the same group time by method in embodiment 3, result is as follows:
| Sequence number | Group | Envelop rate | |
| 1 | SL | (63.05?±2.04)% | |
| 2 | ST | (68.67?±0.59)% | |
| 3 | NT-28 | (66.45?±6.00)% | |
| 4 | NT-46 | (66.87?±3.78)% | |
| 5 | NT-64 | (78.98.21?±2.94)% | |
| 6 | NT-82 | (65.53?±2.77)% | |
| 7 | DT | (74.345?±2.74)% |
Result shows, no matter which kind of Lipid composition, the envelop rate of carrier all has raising in various degree compared with SL group.Wherein the raising of NT-64 group is the most obvious, and the ST group of its more single phospholipid carrier is also significantly increased with DT group, and all the other composite phospholipid carriers fail to fully demonstrate the advantage of medicine carrying effect aspect.
(2) morphology evaluation
Adopt transmission electron microscope to carry out morphological observation to preparing gained SL, ST, NT-64 and DT, concrete process of the test is as follows: liposome and carrier are diluted to after suitable concentration with phosphate buffer (PBS) respectively, get and drop in right amount on microscope slide, online to drop to special-purpose copper year after 1% phosphotungstic acid negative staining, naturally volatilize afterwards with transmission electron microscope observation and take pictures, the results are shown in Figure 4.Visible all particles are the spherical vesicle of class, structure rounding, and peripheral special finger print lipid bilayer is clear and legible.
(3) particle diameter and distribution thereof
Adopt Zeta PALS type high-resolution Zeta potential and Particle Size Analyzer, take PBS as disperse medium, measure mean diameter (Particle size), half-peak breadth (Half width) and the polydispersity coefficient (Polydispersity, PI) of liposome and carrier.Before practical measurement, testing sample need be diluted to counting rate (Count Rate) with PBS and be no more than 500 Kcps to guarantee the accuracy of measurement result.Every duplicate samples parallel assay 3 times, acquired results sees the following form.
Particle diameter and the distribution (n=3, x ± s) thereof of different Lipid composition Propranolol carriers
Above result shows, the particle size distribution of the Propranolol carrier of all Lipid compositions is homogeneous (polydispersity coefficient is all less than 0.3) comparatively, compares particle diameter slightly increase with SL.The mean diameter of the NT of different Lipid compositions is basically identical, and compare with ST slightly and increase, but all than little at least 20 nm of DT.
(4) Zeta potential evaluation
Adopt Zeta PALS type high-resolution Zeta potential and Particle Size Analyzer, take ultra-pure water as disperse medium, measure the Zeta potential of liposome and carrier, every duplicate samples parallel assay 3 times, acquired results sees the following form.
The Zeta current potential (n=3, x ± s) of different Lipid composition Propranolol carriers
| Sequence number | Group | Zeta potential | |
| 1 | SL | -（2.4?±0.04） | |
| 2 | ST | -（3.1?±0.03） | |
| 3 | NT-28 | -（3.6?±0.09） | |
| 4 | NT-46 | -（2.1?±0.07） | |
| 5 | NT-64 | -（3.4?±0.12） | |
| 6 | NT-82 | -（3.9?±0.09） | |
| 7 | DT | -（4.4?±0.10） |
As seen from the above table, the Propranolol carrier of all Lipid compositions is all nearly neutrality.
(5) morphotropism evaluation
Morphotropism is the unique physicochemical properties of carrier, in classical osmotic gradient and Hydration Force theoretical explanation, the deformability of carrier is role with pharmaceutical carrier for it, carry active medicine distortion by the special lipid passage in horn cell gap, thereby promote that percutaneous drug absorption plays vital effect, therefore the important indicator that Chang Zuowei carrier character is evaluated.
Carrier penetrates horny layer and arrives in the process of deep skin tissue in distortion, and along with the variations in temperature of different depth skin, the mobility of carrier lipid bilayer may change, and directly translates into the variation of its deformability.Therefore this test adopts 32 ℃ of skin superficial temperature and 37 ℃ of temperature of extruding as carrier microporous filter membrane of deep skin tissue temperature, and gained morphotropism index is designated as respectively to DI32, DI37, to realizing, evaluate more all sidedly the morphotropism that may the exist variation of carrier in actual percutaneous absorption process.
Concrete process of the test is as follows: get carrier suspension 1 mL, with PBS, be diluted to and be injected into liposome after 10 mL and extrude in instrument, with 0.5 MPa nitrogen pressure, extrude, and extrude omnidistance with super constant temperature water tank control the temperature of extruding be respectively 32 ℃ with 37 ℃.Record carrier distortion in 5 min and, by the volume of 100 nm aperture polycarbonate membranes, measure in time the carrier particle diameter after extruding, by following formula, ask respectively the morphotropism index of calculating the carrier under different extrusion temperatures:
DI?=J?×（rv?/rp）2
In above formula, the morphotropism index that DI is carrier; J(mL) be the volume of extruding of carrier; Rv(nm) be the carrier particle diameter of microporous filter membrane after extruding; Rp(nm) be microporous filter membrane aperture.
Then ask and calculate the morphotropism situation of change that Δ DI=DI37-DI32 causes because of temperature contrast to describe different Lipid composition Propranolol carriers, parallel three parts of above-mentioned test, the results are shown in Figure 5.
In Fig. 5, experimental result shows, the Δ DI value of all kinds composite phospholipid carrier is compared with single phospholipid carrier, all there is remarkable increase, wherein obvious with the amplification of NT-64 again, infer that NT rises to from 32 ℃ in temperature the process of 37 ℃, in lipid bilayer may there is certain variation in the conformation of phospholipid and the mobility of lipid film, and as the mobility increase of the conformation upset of phospholipid and lipid film, and this variation is directly cashed as the increase of carrier morphotropism.Otherwise ST, DT are in 32 ℃ ~ 37 ℃ temperature fluctuation ranges, in lipid bilayer, the conformation of phospholipid and the mobility variations of lipid film are not remarkable.
In summary, compare with single phospholipid carrier, composite phospholipid carrier embodies the feature that morphotropism increases in 32 ℃ ~ 37 ℃ these temperature uphill process, may make it when actual administration, shows the difference of pharmacokinetic property and pharmacological activity aspect.
(6) estimation of stability (leak test)
According to the result of aforementioned pharmaceutical properties comparison, choose SL and ST, these 3 kinds of carriers of NT-64, DT are carried out leak test, and investigated respectively under 37 ℃ of water bath condition, the leakage situation of Propranolol when adding rat plasma and not adding rat plasma, thus probe into Lipid composition for the impact of medicine carrying carrier stability.
(1) Propranolol carrier 0.5 mL is got in the leakage that does not add rat plasma, sephadex column is removed free drug and is collected pastille part 5 mL, all be placed in bag filter, after tightening, using the PBS of 50 mL volumes as accepting medium, 500 rmin under 37 ℃ of water bath condition
-1magnetic agitation, and respectively at accurate acceptable solution 0.5 mL, 12000 rmin of drawing of 0,15,30,60,120,180,240,360,480,720 min
-1after centrifugal 10 min, HPLC measures drug level, supplies in time the fresh accepting medium of equal-volume after every sub-sampling.
By following formula, asked the accumulation leakage rate L(μ g that calculates different time points medicine).
L?=V?×Cn?+?V0∑Ci
In formula, V is acceptable solution volume (mL); V0 is sample volume (mL); Cn is acceptable solution Chinese medicine concentration (the μ gmL that n sampling time point records
-1); Ci is acceptable solution Chinese medicine concentration (the μ gmL that the individual sampling time point of i (i≤n-1) records
-1);
Propranolol total amount in leak test front transfer body is counted to 100%, by following formula, ask the drug accumulation of calculating each time point to leak percentage rate L%(%).
L%?=L/LT?×100%
In above formula, LT(μ g) for leaking the total amount of Propranolol in front bag filter.
More than test parallel 3 parts, with L% to time t(min) mapping, obtain L%-t curve and see Fig. 6.
Result of the test shows as shown in Figure 6, now the leakage of ST is the most serious, the L% of 720 min is the more than 1.5 times of SL, adding of prompting edge activating agent, when improving the mobility of carrier lipid film and giving its special deformability, also produced the problem that membrane permeability increase, entrapped drug easily leak.And this problem for need to be with the functional form of carrier medicament-carried angle of penetration cell plastid gap lipid passage and promote for the carrier of percutaneous drug absorption, be totally unfavorable.The leakage of medicine probably has a strong impact on the performance of carrier function vector in transdermal administration process, thereby cannot promote to embody advantage aspect percutaneous drug absorption.
Now, the stability of DT is best, the L% less than 20% of 720 min; Yet when rat plasma exists, its leak rate before 240 min has been compared obvious increase when not adding rat plasma, and the leak rate of front 120 min even will be higher than ST.Prompting: the composition in rat plasma may interaction (lipid exchange or fusion etc.) occur with DT lipid film, the mobility and the permeability that have become its lipid film have been changed, cause drug leakage to be accelerated, but this interaction does not fundamentally destroy the basic structure (after 240 min, its leakage tends to be steady) of DT lipid bilayer.
(2) the leak test method that adds rat plasma is substantially the same, detailed process is as follows: get Propranolol carrier 1 mL, sephadex column is removed free drug and is collected pastille part 5 mL, getting 2.5 mL and equal-volume rat plasma mixes and is placed in bag filter, after tightening, using the PBS of 50 mL volumes as accepting medium, 500 rmin under 37 ℃ of water bath condition
-1magnetic agitation, and respectively at accurate acceptable solution 0.5 mL, 12000 rmin of drawing of 0,15,30,60,120,180,240,360,480,720 min
-1after centrifugal 10 min, HPLC measures drug level, supplies in time the fresh accepting medium of equal-volume after every sub-sampling.Correlation method for data processing process is constant, and acquired results is shown in Fig. 7.
In Fig. 7, result shows, NT-64 all shows good stability in above two kinds of leak tests, and especially when rat plasma exists, its leakage is minimum, may be relevant with the special nature of its composite phospholipid.Splendid deformability in conjunction with it 37 ℃ time, while inferring 37 ℃ of left and right, may there is near the critical state (being that phase transition temperature is 37 ℃) changing mutually in NT-64, and this critical state may be given the special pharmaceutical properties of NT-64.
In the body of embodiment 7 Propranolol composite phospholipid carriers, blood drug level and blood drug level-area under curve (AUC) are evaluated
(1) specific experiment step is as follows:
1. test grouping: pharmacokinetics test grouping and code thereof see the following form:
2. before administration, prepare: transdermal administration group rat need be in anesthesia the previous day on-test, the depilation of abdominal part machinery, can carry out pharmacokinetics test after recovering to spend the night.Intravenous injection group is processed without depilation.In addition, before the administration of all experimental group rats, equal fasting 12 h, can't help water.
3. administration and blood sampling: the dosage of all experimental grouies is 8 mgkg
-1, the administration area of transdermal administration group is 4 * 4 cm
2.The concrete grammar of transdermal administration is as follows: rat anesthesia layback position is fixed on Mus plate, marks 4 * 4 cm
2administration area, test sample is evenly distributed on administration skin with pipettor, rat is positioned over separately in mouse cage to predetermined point of time blood sampling, 12000 rmin under-4 ℃ of conditions after volatilizing
-1centrifugal 1 min, divides and gets blood plasma, and-20 ℃ frozen to be measured.
4. plasma sample is processed: the accurate rat plasma 100 μ L that draw are to clean tool plug scale test tube, and (mass concentration is 1000 ngmL to add the carvedilol methanol solution of 1/10 blood plasma volume
-1) and the saturated sodium carbonate solution of 1/5 blood plasma volume, after vortex mixed 30 s, then add 1 mL normal hexane-dichloromethane-isopropyl alcohol (65:30:5), after vortex mixed 3 min, 4000 rmin
-1centrifugal 10 min, draw the supernatant.Residue repeats above alkalization-extracting operation once, and after combining extraction liquid, under 50 ℃ of water-baths, nitrogen current dries up, and residue obtained use 100 μ L mobile phases are redissolved, 12000 rmin under-4 ℃ of conditions
-1centrifugal 10 min, get the analysis of supernatant sample introduction.
5. high-efficient liquid phase chromatogram technique analysis plasma sample: the sample after above-mentioned processing is analyzed according to high-efficient liquid phase chromatogram condition sample introduction described in embodiment 3.
The plasma sample of processing method Treatment Analysis has good stability thus, and completely separated, the endogenous impurity in blood plasma does not disturb the assay of Propranolol, sees Fig. 8.
(2) blood drug level and blood drug level-area under curve (AUC) are evaluated
Plasma sample room temperature is processed and sample introduction according to said method after thawing, and determines according to actual needs and according to corresponding standard curve, calculates sampling volume Propranolol blood drug level, and draw blood drug level-time graph as shown in Figure 9.By DAS 2.0 software processes for the data obtained, calculate pharmacokinetic parameter (statistical moment parameter) and see the following form again.
The pharmacokinetic parameter comparison of each test group (n=5, x ± s)
Note: in table, alphabetical implication is AUC: blood drug level-area under curve; MRT: average residence time in body;
T
1/2: biological half-life; Vz: apparent volume of distribution; CLz: clearance rate;
T
max: peak reaching time of blood concentration; C
max: blood drug level peak concentration
According to above result of the test, draw to draw a conclusion:
NT-64 shows obvious percutaneous and absorbs advantage, its AUC (0-∞) is nearly 2.5 times of S-i.v. group, nearly 5 times of S-t.d. group, MRT (0-∞) is the longest in all test group, Cmax compares and all has utmost point significant difference (P is all less than 0.01) with other transdermal administration groups, but its peak time (4 h) is compared prolongation to some extent with other two kinds single phospholipid carriers (3 h).
S-t.d. the AUC (0-∞) of group approaches 50% of S-i.v. group, during prompting Propranolol solution transdermal administration, can bring into play drug depot effect.
ST percutaneous absorbs not good, and Cmax, AUC (0-∞) are minimum in all experimental grouies, and this may leak in transdermal process the poor stability of drug encapsulation, medicine with it, thereby while hindering its transdermal administration, the performance of function vector is relevant.
The percutaneous absorption performance of DT is good compared with ST, and its AUC (0-∞) is nearly 3 times of ST, but the evaluation result contradiction of this result and morphotropism.
SL also shows the effect that good promotion Propranolol percutaneous absorbs, and its peak time is only that 2 h show that liposome promotes the mechanism of water soluble drug percutaneous absorption to compare difference to some extent with carrier.
The percutaneous absorption advantage of NT-64 is related to the special nature that NT-64 shows in pharmaceutics test, infers: composite phospholipid carrier has formed certain cenotype when DPPC and SPC mol ratio are 6:4, and the phase transition temperature of this cenotype is probably near 37 ℃.Exactly because and have the critical state changing mutually 37 ℃ time, thereby make NT-64 show unique pharmaceutical properties and pharmacokinetic characteristic.Its main manifestations is for improving Propranolol envelop rate, strengthening the stability that Propranolol is sealed, and in the time of 37 ℃ special deformability, and the pharmacokinetic property while therefore causing its transdermal administration improves, bioavailability improves.
The mensuration of embodiment 8 Propranolol composite phospholipid carrier phase transition temperatures
By method described in embodiment 2, prepare Propranolol composite phospholipid carrier, wherein consumption is DPPC:SPC=6:4, tests its phase transition temperature, and test condition is as follows:
Test instrunment: Diamond DSC differential scanning calorimeter, Pyris Series-Diamond DSC data analysis software; Crucible type: standard stainless steel crucible, seal cover; Amount of samples: 50 mg; DSC heating schedule: 1 ℃ of min
-1; Atmosphere: high pure nitrogen; Flow velocity: 50 cm
3min
-1;
Test result is shown in Figure 10, result from figure, and the composite phospholipid in carrier merges and becomes a cenotype at about 37 ℃.
Claims (8)
1. the Propranolol composite phospholipid carrier that can promote percutaneous drug absorption, it is characterized in that it is that by two class phospholipid composition dipalmitoyl phosphatidyl choline and soybean phospholipid, edge activating agent and Propranolol, to press row weight portion proportioning made: 1 ~ 10 part of dipalmitoyl phosphatidyl choline, 1 ~ 10 part of soybean phospholipid, 0.2 ~ 2 part of edge activating agent, the amount of Propranolol is 0.05 ~ 0.5 times of phospholipid total amount; Wherein edge activating agent is selected from sodium cholate, NaTDC, sorbester p17 and polysorbas20.
2. composite phospholipid carrier according to claim 1, is characterized in that: described dipalmitoyl phosphatidyl choline equals 8:2 to 2:8 with the mole ratio of soybean phospholipid.
3. composite phospholipid carrier according to claim 1, is characterized in that: described dipalmitoyl phosphatidyl choline equals 6:4 with the mole ratio of soybean phospholipid.
4. composite phospholipid carrier according to claim 1, is characterized in that: described edge activating agent is Arlacel-80.
5. a method of preparing the Propranolol composite phospholipid carrier described in any one in claim 1 ~ 4, is divided into the blank composite phospholipid carrier of preparation and the large step of medicine carrying two, and its concrete operation step is as follows:
(1) take by weight 1 ~ 10 part of dipalmitoyl phosphatidyl choline, 1 ~ 10 part of soybean phospholipid, 0.2 ~ 2 part of edge activating agent, is dissolved in dehydrated alcohol, injects ammonium sulfate solution;
(2) by the method heating or reduce pressure, remove ethanol;
(3) supersound process or high pressure breast are even, reduce the particle diameter of composite phospholipid carrier;
(4) by dialysis, ultrafiltration or gel filtration chromatography method, remove the ammonium sulfate of not sealed by composite phospholipid carrier, form blank composite phospholipid carrier suspension;
(5) getting total amount is that the Propranolol of 0.05 ~ 0.5 times of phospholipid total amount adds temperature in above-mentioned blank composite phospholipid carrier suspension to incubate, and it is sealed by composite phospholipid carrier, makes Propranolol composite phospholipid carrier.
6. preparation method according to claim 5, is characterized in that: described in step (1) be dissolved in dehydrated alcohol time, with the speed of 5 ml/min, inject the ammonium sulfate solution of 0.1 ~ 0.3 mol/L of 10 ~ 20 times of volumes.
7. according to the preparation method described in claim 5, it is characterized in that: in step (4), adopt the phosphate buffer of pH7.4 to dialyse to remove the ammonium sulfate of not sealed by composite phospholipid carrier.
8. preparation method according to claim 5, is characterized in that: it is 0 ~ 60 ℃ that the temperature described in step (5) is incubated temperature, and the temperature time of incubating is 5 ~ 60 min.
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Non-Patent Citations (6)
| Title |
|---|
| Pandey Shivanand等.Transferosomes: A Novel Approach for Transdermal Drug Delivery.《Der Pharmacia Lettre》.2009,第1卷(第2期),144-150. |
| Transferosomes: A Novel Approach for Transdermal Drug Delivery;Pandey Shivanand等;《Der Pharmacia Lettre》;20091231;第1卷(第2期);144-150 * |
| 吴佳君等.普萘洛尔治疗婴幼儿血管瘤的新进展.《中国美容医学》.2011,第20卷(第1期),163-165. |
| 布洛芬传递体的制备及体外透皮研究;郭伟英;《中国医药工业杂志》;20101231;第41卷(第3期);191-193 * |
| 普萘洛尔治疗婴幼儿血管瘤的新进展;吴佳君等;《中国美容医学》;20110131;第20卷(第1期);163-165 * |
| 郭伟英.布洛芬传递体的制备及体外透皮研究.《中国医药工业杂志》.2010,第41卷(第3期),191-193. |
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