CN107569447B - Soluble microneedle patch loaded with tuberculin pure protein derivative and preparation method thereof - Google Patents
Soluble microneedle patch loaded with tuberculin pure protein derivative and preparation method thereof Download PDFInfo
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Abstract
The invention relates to a soluble microneedle patch loaded with tuberculin pure protein derivatives and a preparation method thereof, wherein the soluble microneedle patch comprises a substrate and a needle body, wherein the substrate is prepared from a first high polymer material; the needle body is prepared from a conjugated rhzomorph pure protein derivative and a needle body forming material; the pure protein derivative of the conjugated rhzomorph accounts for 0.04 to 0.10 percent of the total weight of the needle body; the needle body shaping material comprises a second high polymer material and a small molecule material; the second high polymer material is selected from at least one of hyaluronic acid or sodium salt thereof, hydroxyethyl starch, polyvinylpyrrolidone and polyethylene glycol; the small molecular material is selected from at least one of water-soluble small molecular sugar, acidic water-soluble amino acid and acidic water-soluble salt; the mass ratio of the second high molecular material to the small molecular material is 3-8: 1. The soluble microneedle patch has good puncture strength and in-vivo dissolution speed.
Description
Technical Field
The invention relates to the technical field of pharmaceutical preparations, in particular to a soluble microneedle patch loaded with tuberculin pure protein derivatives and a preparation method thereof.
Background
The 2009 world health organization global tuberculosis survey report shows that the number of newly-added combined patients in the world is 927 thousands every year, China is one of tuberculosis high-load countries, the number of deaths caused by tuberculosis every year is 20 thousands, and the prevention and control situation is still very severe. Early diagnosis of tuberculosis is a key link and factor for controlling tuberculosis spread, so that the early simple diagnosis technology determines the discovery rate level of tuberculosis patients, and the current primary diagnosis of tuberculosis mainly depends on the skin test experiment of conjugate and subsequent technologies such as imaging, etiology, serum antibody detection and the like.
The tuberculin test is used as an auxiliary means for diagnosing initial tuberculosis, and plays an important role in clinical diagnosis and tuberculosis epidemiological investigation. With the continuous development of new medical technology, the new generation of tuberculin pure protein derivative (TB-PPD) has high purity and strong specificity, and if the tuberculin test is combined with other conditions of a subject, the tuberculin test can still help the early diagnosis of tuberculosis, help epidemiologists to discover infectious and susceptible people of tubercle bacillus in people, even discover the infectious source in specific people. Especially, the tuberculin test is developed in people under 15 years old, and the tuberculin test has important significance for guiding the bcg replanting of children and the preventive medication and treatment of infected people. Tuberculin test has been used as a conventional test means, and various methods such as a skin-scarification method (Pirquet method), an ointment paste method (Moro method), a skin-prick method (Heaf method) and an intradermal injection method (Mantoux method) have been inoculated, and the former 3 methods are rarely used because the dose is not easily standardized, and the measurement result is difficult to judge. At present, intradermal injection is mainly used, the method is accurate in dosage, test results are easy to judge, but professional technicians and technical requirements are required to be strict, the injection depth and angle determine the results of a skin test experiment, and certain misdiagnosis risk exists. Meanwhile, the group dependence of children and infants is relatively poor due to injection pain, and some adverse reactions are occasionally caused by intradermal injection, such as experimental scar suppurative infection, lymphangitis and the like which are caused by improper nursing after injection in clinic. On the other hand, whether the safety of the sterile needle is guaranteed or not, whether the hidden danger of repeated cross infection exists or not, the cost of the needle and a plastic pipe material is high, the environment pollution is serious during production and manufacturing, the factors of difficult subsequent treatment and the like become the defect of using intradermal injection for administration after use, and under the influence of various adverse factors, the skin test diagnosis method which is safe and effective and has high compliance is the most critical problem at present.
The soluble micro-needle percutaneous immunization approach is taken as a novel percutaneous immunization method, and the time from the proposal to the continuous development is decades, and the method is characterized by convenient use, easy operation of common personnel, and simultaneously, because the length of a micro-needle body is limited, the micro-needle body can not touch skin nerves when being pierced, no pain exists, the compliance of special population is good, and the needle point of the medicine carrying just reaches the cortex part with the most immune cells under the condition of fully penetrating the skin cuticle, the success rate of the experiment is high, and the clinical diagnosis is more convincing. And especially the unique advantages of the soluble micro-needle, the problems of secondary damage, biocompatibility and the like of the traditional silicon and metal micro-needle are solved, the defects of leakage, large dosage difference and the like of the injection micro-needle are avoided, and the injection micro-needle has the advantages of convenient production and preparation, relatively large drug loading rate and the like. However, the needle bodies of most of the prior soluble microneedles have the defects of poor rigidity, low skin penetration rate and the like, and the intracutaneous delivery is difficult to realize effectively; the needle body of the existing soluble microneedle has low swelling and dissolving speed in the skin and slow drug release, can be used for carrying long-acting treatment medicaments, and can be used for observing the reaction condition of the skin after skin test medicaments such as tuberculin medicaments are required to be quickly used, so that the requirements on the dissolving speed of the microneedle penetrating into the skin and the release speed of the medicaments are higher, and the existing soluble microneedle is difficult to meet the requirements.
Disclosure of Invention
Based on the soluble microneedle patch, the invention provides the soluble microneedle patch loaded with the tuberculin pure protein derivative, and the soluble microneedle patch has good puncture strength and in-vivo dissolution speed.
The specific technical scheme is as follows:
a soluble microneedle patch carrying tuberculin pure protein derivative comprises a substrate and a needle body, wherein the substrate is prepared from a first high polymer material; the needle body is prepared from a conjugated rhzomorph pure protein derivative and a needle body shaping material, wherein the percentage of the conjugated rhzomorph pure protein derivative in the total weight of the needle body is 0.04-0.10%; the needle body shaping material comprises a second high polymer material and a small molecule material; the second high polymer material is selected from at least one of hyaluronic acid or sodium salt thereof, hydroxyethyl starch, polyvinylpyrrolidone and polyethylene glycol; the small molecular material is selected from at least one of water-soluble small molecular sugar, acidic water-soluble amino acid and acidic water-soluble salt; the mass ratio of the second high molecular material to the small molecular material is 3-8: 1.
In some of these embodiments, the water-soluble small molecule sugar is selected from at least one of glucosamine, glucose, and mannitol; the water-soluble amino acid with partial acidity is at least one of glutamic acid and aspartic acid; the water-soluble salt with meta-acidity is at least one selected from sodium dihydrogen carbonate and sodium dihydrogen phosphate.
In some embodiments, the mass ratio of the second polymer material to the small molecule material is 4-6: 1.
In some embodiments, the pure protein derivative of the conjugate is present in an amount of 0.07% to 0.085% by weight of the needle body.
In some embodiments, the needle body forming material comprises the following components in parts by weight:
in some embodiments, the needle body forming material comprises the following components in parts by weight:
in some of these embodiments, the hyaluronic acid or sodium salt thereof has an average molecular weight of less than 10000, and the hydroxyethyl starch has an average molecular weight of 15000-50000; the average molecular weight of the polyvinylpyrrolidone is 10000-70000; the average molecular weight of the polyethylene glycol is 300-800.
In some of the examples, the hyaluronic acid or its sodium salt has an average molecular weight of 4000-.
In some of these embodiments, the first polymeric material is selected from: at least two of hydroxypropyl cellulose, hydroxypropyl methylcellulose, hyaluronic acid or its sodium salt, and glycerol; the hydroxypropyl cellulose satisfies the following conditions: when the concentration of the aqueous solution of hydroxypropyl cellulose is 10 wt%, the viscosity of the aqueous solution of hydroxypropyl cellulose is 50-750 mPa.s; the hydroxypropyl methylcellulose satisfies the following conditions: when the concentration of the aqueous solution of hydroxypropyl methylcellulose is 10 wt%, the viscosity of the aqueous solution of hydroxypropyl methylcellulose is 3-150 mpa.s; the hyaluronic acid or sodium salt has an average molecular weight of not less than 200 ten thousand.
In some of these embodiments, the hydroxypropyl cellulose satisfies the following condition: when the concentration of the aqueous solution of hydroxypropyl cellulose is 10 wt%, the viscosity of the aqueous solution of hydroxypropyl cellulose is 150-450 mPa.s; the hydroxypropyl methylcellulose satisfies the following conditions: when the concentration of the aqueous solution of hydroxypropyl methylcellulose is 10 wt%, the viscosity of the aqueous solution of hydroxypropyl methylcellulose is 5-50 mPa.s; the average molecular weight of the hyaluronic acid or the sodium salt is 200-2000 ten thousand.
In some embodiments, the first polymer material is composed of the following components in parts by weight:
5-15 parts of hydroxypropyl methyl cellulose
10-20 parts of hydroxypropyl cellulose
Hyaluronic acid or sodium salt 0.1-0.8 parts.
In some embodiments, the first polymer material is prepared from the following components in parts by weight:
7-9 parts of hydroxypropyl methyl cellulose
14-16 parts of hydroxypropyl cellulose
Hyaluronic acid or sodium salt 0.2-0.4 parts.
In some of these embodiments, the needle body of the soluble microneedle loaded with purified protein derivative of tuberculin has a height of 200-500 um.
In some embodiments, the diameter of the bottom of the needle body of the soluble microneedle loaded with the tuberculin pure protein derivative is 150-200um, and the diameter of the needle tip of the needle body is 1-40 um.
The invention also discloses a preparation method of the soluble microneedle patch loaded with the tuberculin pure protein derivative.
The specific technical scheme is as follows:
the preparation method of the soluble microneedle patch loaded with the tuberculin pure protein derivative comprises the following steps:
(1) preparing a needle body: dissolving the second high molecular material, the small molecular material and the tuberculin pure protein derivative by using water to obtain a needle body solution; injecting the needle body solution into a microneedle female die, pressurizing to enable the needle body solution to fill needle point micropores of the microneedle female die, removing redundant needle body solution, and drying;
(2) preparation of the substrate: and (2) dissolving the first high polymer material with water to serve as a substrate solution, uniformly coating the substrate solution in the microneedle female die treated in the step (1), pressurizing, drying and demolding to obtain the microneedle patch carrying the tuberculin pure protein derivative.
In some embodiments, the total concentration of the second polymeric material and the small molecule material in the needle solution is 25-32 wt%.
In some of these embodiments, the concentration of the first polymeric material in the base solution is 18 to 25 wt%.
In some of these embodiments, the pressurizing in the needle body preparing step comprises: pressurizing the whole microneedle female die in an environment with the pressure of 4.5-5.5 atm for 4-6min, wherein the drying conditions comprise: drying at 20-30 deg.C under normal pressure for 1.5-2.5 hr.
In some of these embodiments, the pressurizing in the step of preparing the substrate comprises: placing the whole microneedle female die in an environment with the pressure of 0.8-1.2 atmospheric pressure, and pressurizing for 9-11s, wherein the drying conditions comprise: drying at 20-30 deg.C under normal pressure for 5-7 hr.
The soluble microneedle patch carrying the tuberculin pure protein derivative and the preparation method thereof have the following advantages and beneficial effects:
the soluble microneedle patch loaded with the tuberculin pure protein derivative is characterized in that a proper high-molecular material and a small-molecular material are selected to be matched in a certain proportion, so that a prepared microneedle body has good hardness and skin puncture strength, the selected small-molecular material can adjust the dissolving speed of the microneedle body in alkalescent tissue liquid after entering the skin, the soluble small-molecular material and body fluid generate interaction when macromolecules of the microneedle body swell and dissolve in water, the dissolving of the microneedle in the body is accelerated, and the requirement of a skin test experiment on the dissolving speed of drugs can be met.
In the existing soluble microneedle patch, the basal layer part of the soluble microneedle patch is large in brittleness, hard and brittle after being dried by high polymer materials (such as PVP, PVA and other materials), but generally the skin is an elastic tissue, and is easy to dent when being pressed by external force, so that the hard and brittle basal layer is easy to crack in the pressing process, the drug transfer effect of the microneedle is seriously influenced, and meanwhile, the sticking feeling is poor, and the soluble microneedle patch is difficult to accept by people. The invention further optimizes the high polymer material for preparing the substrate, so that the substrate layer of the prepared soluble microneedle patch has good flexibility, can be bent to a higher degree without influencing the needle body of the microneedle, avoids the breakage of the microneedle due to external force and improves the compliance attached to the skin.
The invention further optimizes the height of the microneedle body, so that the microneedles are accurately positioned in the skin, and the technical errors caused by the operation errors due to the fact that the needle inserting angle of the intradermal injection needle is emphasized are effectively avoided, so that the misdiagnosis condition of diseases caused by the false positive or false negative results is avoided, and the accuracy of the skin test experiment is improved.
The soluble microneedle patch carrying the tuberculin pure protein derivative can achieve the same diagnosis effect as the original intradermal injection when being tested on the skin of a guinea pig infected by tuberculin, is more favorable for popularization and use, has the advantages of good compliance, no cross infection and the like compared with the intradermal injection mode, and the microneedle displays the medicament in a punctate array form, disperses the action points of the medicament and avoids serious adverse reactions such as blisters, ulcers, local necrosis, lymphangitis and the like of certain special individuals due to overlarge local medicament concentration.
The preparation method of the soluble microneedle patch loaded with the tuberculin pure protein derivative has the advantage of strong operability.
Drawings
FIG. 1 is a microscopic view of a soluble microneedle patch loaded with a purified protein derivative of tuberculin; wherein a is the dissolvable microneedle patch prepared in example 1, B is the dissolvable microneedle patch prepared in example 2, and C is the dissolvable microneedle patch prepared in comparative example 1;
fig. 2 is a hardness test chart of the dissolvable microneedle patch prepared in example 1;
fig. 3 is a hardness test chart of the dissolvable microneedle patch prepared in example 2;
fig. 4 is a hardness test chart of the dissolvable microneedle patch prepared in comparative example 1;
fig. 5 is a skin penetration staining pattern of the microneedles for the puncture performance test of the microneedles in example 3; wherein a is the result of the dissolvable microneedle patch prepared in example 1, and B is the result of the dissolvable microneedle patch prepared in example 2; c is the result of the soluble microneedle patch prepared in comparative example 1;
fig. 6 is the results of the solubility test of the microneedles in example 3; wherein a is the result of the dissolvable microneedle patch prepared in example 1, and B is the result of the dissolvable microneedle patch prepared in example 2; c is the result of the soluble microneedle patch prepared in comparative example 1;
FIG. 7 is the flexibility test results of the substrate layer of example 3; wherein a is the result of the dissolvable microneedle patch prepared in example 1, and B is the result of the dissolvable microneedle patch prepared in example 2; c is the result of the soluble microneedle patch prepared in comparative example 1;
FIG. 8 is a reaction chart of erythema test of example 4; wherein A is the result of a blank control group, and B is the result of a positive control group; c is the result of the blank microneedle control group 1; d is the result of experimental group 1; e is the result of the blank microneedle control group 2; f is the result of experiment group 2.
Detailed Description
The soluble microneedle patch loaded with purified tuberculin protein derivatives and a method for manufacturing the same according to the present invention will be described in further detail with reference to the following examples.
Example 1
The soluble microneedle patch loaded with the purified protein derivative of tuberculin provided by the embodiment is prepared by the following method:
(1) preparing a needle body solution: precisely weighing 0.25g of sodium hyaluronate with average molecular weight of 8000, 0.3g of hydroxyethyl starch with average molecular weight of 40000, 0.1g of polyvinylpyrrolidone with average molecular weight of 12000, 0.1g of glucosamine, 0.03g of glutamic acid, 10mg of sodium dihydrogen phosphate and 2.0ml of 300ug/ml of water solution for injection of the tuberculin pure protein derivative, and mixing, dissolving and swelling.
(2) Preparing a flexible substrate layer solution: accurately weighing 8.0g of hydroxypropyl methylcellulose (the viscosity of 10 wt% aqueous solution is 5mPa.s), 15.0g of hydroxypropyl cellulose (the viscosity of 10 wt% aqueous solution is 300mPa.s), 0.30g of sodium hyaluronate with average molecular weight of 400 ten thousand and 80g of water for injection, and carrying out vortex mixing, uniform mixing and dissolution.
(3) Preparing a pure protein derivative soluble microneedle patch carrying the conjugated rhzomorph:
a) injecting the prepared needle body solution into a PDMS (polydimethylsiloxane) microneedle female die with the height of a microneedle body of 400um, the diameter of the bottom of the microneedle body of 200um and the diameter of a needle point of 15-30um, putting the whole die into an environment with 5 atmospheres, pressurizing for 5min, filling the needle body solution into the needle point micropores, removing the redundant needle body solution, and then placing the whole die into a dryer (25 ℃, normal pressure) for drying for 2 h.
b) Uniformly coating the flexible basal layer solution on the surface of the microneedle female die treated in the step a), quickly placing the microneedle female die in an environment with 1 atmospheric pressure, instantly pressurizing for 10s, taking out, drying for 6 hours, and demolding to obtain the soluble microneedle patch carrying the tuberculin pure protein derivative. The microscopic view is shown in fig. 1, panel a.
Example 2
The soluble microneedle patch loaded with the purified protein derivative of tuberculin provided by the embodiment is prepared by the following method:
(1) preparing a needle body solution: precisely weighing 0.3g of sodium hyaluronate with average molecular weight of 8000, 0.15g of hydroxyethyl starch with average molecular weight of 20000, 0.15g of polyvinylpyrrolidone with average molecular weight of 30000, 0.15g of glutamic acid, 8mg of sodium dihydrogen phosphate and 2.0ml of water solution for injection of 600ug/ml tuberculin pure protein derivative, mixing, dissolving and swelling.
(2) Preparation of flexible substrate layer solution: 20.0g of hydroxypropyl methyl cellulose (the viscosity of a 10 wt% aqueous solution is 5mPa.s), 1.0g of sodium hyaluronate with the average molecular weight of 200 ten thousand and 80g of water for injection are precisely weighed, vortexed, uniformly mixed and dissolved.
(3) Preparing a pure protein derivative soluble microneedle patch carrying the conjugated rhzomorph: the same as in example 1. The microscopic view is shown in fig. 1, panel B.
Comparative example 1
The comparative example provides a preparation method of a dextran soluble microneedle patch loaded with tuberculin pure protein derivatives, and the preparation method comprises the following steps:
(1) preparation of needle body solution
0.75g dextran (molecular weight 70000) and 2.0ml of 300ug/ml tuberculin pure protein derivative injection water solution are weighed, mixed, dissolved and swelled to obtain the needle body solution with the excipient being the dextran 70000.
(2) Preparation of base layer solution
Weighing 25g of polyvinylpyrrolidone K90, adding into 85g of absolute ethyl alcohol, and performing vortex swelling and dissolution to obtain a polyvinylpyrrolidone K90 basal layer solution.
(3) Preparation of soluble microneedles
Injecting the solution of the needle body into a female mold of a microneedle with the height of the microneedle body of 400um, centrifuging at 4000rpm for 10min, scraping the redundant solution of the needle point, then adding the solution of the base layer, centrifuging at 4000rpm for 5min again, placing the female mold of the microneedle into a normal-temperature drying box, drying at normal temperature for 12 hours, and taking the microneedle out of the female mold of the microneedle to obtain the dextran soluble microneedle. The microscopic view is shown in fig. 1, panel C.
Example 3
The soluble microneedles prepared in examples 1-2 and comparative example 1 were compared in terms of hardness, puncture ability, and solubility, and the specific evaluation and comparison method was performed as follows:
(1) hardness testing of microneedles
The prepared microneedle patch is cut into a 10 multiplied by 10 array of patches (100 needles), the needle point is upwards placed on a horizontal testing platform of a TMS-PRO type food physical property instrument of the American FTC company, an axial vertical force is applied through a P/6 type flat-head stainless steel cylindrical probe at the stable speed of 0.1mm/sec and the excitation force of 0.05N, the measurement parameters are set, the descending speed of the probe is 10mm/min, the compression speed of the probe, the lifting speed of the probe, the compression amount of 90 percent and the acquisition rate of 200 pps/s. The analyzer records the mechanical change during the time the probe contacts the needle tip until a preset amount of compression is reached (microneedle height 400 μm). After the test is finished, taking out the tested microneedle sample on the carrying platform, observing the local morphological change of the microneedle after the action of the probe of the texture analyzer by using a microscope, and comparing the hardness of the microneedle prepared by different materials. The hardness test charts are shown in fig. 2 to 4, respectively.
(2) Puncture performance of microneedles
Removing hair on two sides of a spinal column of a Uratan anesthetized mouse, slightly lifting back skin, placing on an objective table of a sclerometer, attaching a microneedle patch of a 10 × 10 array on a lower compression column of the sclerometer, quickly pressing down the microneedle patch to penetrate into the skin, stopping pressing down when a pressure gauge of the sclerometer reaches 3kg (30N), keeping the lower compression column on the surface of the skin for 2min, slowly lifting the lower compression column of the sclerometer after the time reaches 2min, peeling the microneedle patch from the surface layer of the skin, then placing the back skin of the mouse under a micro microscope to observe under a full light lamp, collecting images before and after the skin surface layer is penetrated, rapidly and uniformly smearing the back skin of the mouse treated by the microneedle by using 0.4% trypan blue staining solution after the collection is finished, placing for 2-3 min, washing off the trypan epidermal layer by clear water, placing the skin under the micro microscope to observe, before and after the skin surface layer was punctured, staining of living cells with trypan blue was observed, and an image was collected (as shown in fig. 5), and the number of stained spots was counted to calculate the percentage of skin puncture.
The puncture percentage is the number of punctured micropores/the number of the whole microneedle multiplied by 100%
(3) Solubility of microneedles
Unhairing two sides of the spinal column of the back of a urethane anesthetized mouse, slightly lifting the skin of the back, sticking a microneedle patch with an adhesive tape on the unhaired area by hand, pressing for 5min, sticking the adhesive tape on the surface of the skin, fixing the microneedle on the skin, peeling off the microneedle after 20min, cutting the microneedle into a row arrangement by using a hard blade, measuring the height of the residual microneedle by using a micro microscope, and calculating the solubility of different microneedles in the skin at 20min (the dissolution condition is shown in figure 6).
Percent dissolution in skin ═ total microneedle height-actual height measured)/total microneedle height × 100%
(4) Flexibility of the substrate layer
The soluble microneedle patch was gripped with forceps, bent by force at 90 °, and then the degree of disintegration of the basal layer was observed (the result is shown in fig. 7).
And (3) testing results:
example 4
Tuberculin test examination was conducted on the soluble microneedle patches loaded with tuberculin pure protein derivatives prepared in examples 1 and 2 using guinea pigs as the subjects.
Considering that the preparation requirement of the infectivity combined animal test model is strict and the infection risk exists, the common inactivated standard BCG vaccine is adopted as the experimental medicine for molding, and the guinea pig is adopted as the molding object.
Experimental animals and groups: guinea pigs and females weighing 320g ± 30g and 18 in total were divided into 6 groups of 3, and the groups were sequentially divided into a tuberculin pure protein derivative positive injection group (designated as a positive control group), a saline blank injection group (designated as a blank control group), a blank microneedle group prepared by the method of example 1 (containing no bindin pure protein derivative and designated as a blank microneedle control group 1), a blank microneedle group prepared by the method of example 2 (containing no bindin pure protein derivative and designated as a blank microneedle control group 2), a microneedle patch group prepared in example 1 (designated as an experimental group 1), and a microneedle patch group prepared in example 2 (designated as an experimental group 2).
Experimental materials:
bacillus Calmette-Guerin (BCG) injected into skin with specification of 0.5mg, Chengdu biological products research institute, Inc., viable count of Bacillus Calmette-Guerin per 1mg should not be less than 1.0 × 106CFU。
The experimental method comprises the following steps:
(1) guinea pig bacillus calmette-guerin vaccination: taking healthy guinea pig which has been adaptively fed for more than 3 days and is at least 3 months before tuberculin test, injecting BCG vaccine each (BCG)5.0 × 10 into thigh inner skin4And (5) CFU, carrying out artificial experimental modeling.
(2) Pure protein derivative of bindin skin test: in all experimental groups, body hair is removed from two sides of the back spine of the guinea pig through an electric hair remover, 0.1ml of water for injection and a pure protein derivative aqueous solution for conjugate are respectively injected into a blank saline injection group and a positive control group, the other groups respectively press the administration microneedle patch on the back skin of the guinea pig for 5min by hands, then the microneedle patch is fixedly attached to the back skin by using sub-sensitive adhesive cloth, and the patch and the pressure-sensitive adhesive are peeled off in 4 hours.
(3) The mean in-group mean values were calculated by measuring the vertical and horizontal diameters of erythema induration of the skin at 48 and 72 hours, respectively, beginning the time after application (erythema test response after 72 hours is shown in figure 8).
And (4) judging a result: the longitudinal and transverse diameters of the erythema are less than or equal to 5mm and are negative (-), the diameter of 5mm-9mm is general positive (+), and the diameter of 10mm-19mm is moderate positive (+ +).
Results of the experiment
The results show that the soluble microneedle skin test patch loaded with the tuberculin-count pure protein derivative can completely meet the experimental requirements of tuberculin, is simple and easy to operate, is more convenient to popularize, and has better safety and compliance compared with an intradermal injection mode.
The technical features of the embodiments described above may be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the embodiments described above are not described, but should be considered as being within the scope of the present specification as long as there is no contradiction between the combinations of the technical features.
The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.
Claims (10)
1. The soluble microneedle patch loaded with the tuberculin pure protein derivative is characterized by comprising a substrate and a needle body, wherein the substrate is prepared from a first high polymer material; the needle body is prepared from a conjugated rhzomorph pure protein derivative and a needle body forming material; the pure protein derivative of the conjugated rhzomorph accounts for 0.04 to 0.10 percent of the total weight of the needle body;
the needle body forming material comprises the following components in parts by weight:
the average molecular weight of the hyaluronic acid or the sodium salt thereof is less than 10000, and the average molecular weight of the hydroxyethyl starch is 15000-50000; the average molecular weight of the polyvinylpyrrolidone is 10000-70000.
2. The soluble microneedle patch loaded with tuberculin pure protein derivative according to claim 1, wherein the needle body forming material comprises the following components in parts by weight:
3. the soluble microneedle patch carrying a purified protein derivative of tuberculin according to any one of claims 1 to 2, wherein the first polymeric material is selected from the group consisting of: at least two of hydroxypropyl cellulose, hydroxypropyl methylcellulose, hyaluronic acid or its sodium salt, and glycerol; the hydroxypropyl cellulose satisfies the following conditions: when the concentration of the aqueous solution of hydroxypropyl cellulose is 10 wt%, the viscosity of the aqueous solution of hydroxypropyl cellulose is 50-750 mPa.s; the hydroxypropyl methylcellulose satisfies the following conditions: when the concentration of the aqueous solution of hydroxypropyl methylcellulose is 10 wt%, the viscosity of the aqueous solution of hydroxypropyl methylcellulose is 3-150 mpa.s; the hyaluronic acid or sodium salt thereof has an average molecular weight of not less than 200 ten thousand.
4. The soluble microneedle patch loaded with tuberculin pure protein derivative according to claim 3, wherein the first polymer material comprises the following components in parts by weight:
5-15 parts of hydroxypropyl methyl cellulose
10-20 parts of hydroxypropyl cellulose
Hyaluronic acid or its sodium salt 0.1-0.8 parts.
5. The soluble microneedle patch carrying a purified tuberculin protein derivative as set forth in any one of claims 1 to 2, wherein the height of the needle body of the soluble microneedle carrying a purified tuberculin protein derivative is 200-500 μm.
6. A method for preparing the soluble microneedle patch loaded with the purified tuberculin protein derivative according to any one of claims 1 to 5, comprising the steps of:
(1) preparing a needle body: dissolving the needle body excipient and the tuberculin pure protein derivative with water to obtain a needle body solution; injecting the needle body solution into a microneedle female die, pressurizing to enable the needle body solution to fill needle point micropores of the microneedle female die, removing redundant needle body solution, and drying;
(2) preparation of the substrate: and (2) dissolving the first high polymer material with water to serve as a substrate solution, uniformly coating the substrate solution in the microneedle female die treated in the step (1), pressurizing, drying and demolding to obtain the microneedle patch carrying the tuberculin pure protein derivative.
7. The method for preparing a soluble microneedle patch loaded with tuberculin purified protein derivative according to claim 6, wherein the total concentration of the needle excipient in the needle solution is 25-32 wt%.
8. The method for preparing a soluble microneedle patch carrying a purified tuberculin protein derivative according to claim 6, wherein the concentration of the first polymer material in the base solution is 18 to 25 wt%.
9. The method for preparing a soluble microneedle patch carrying a purified tuberculin protein derivative according to any one of claims 6 to 8, wherein the pressurizing in the step of preparing the needle body comprises: pressurizing the whole microneedle female die in an environment with the pressure of 4.5-5.5 atm for 4-6min, wherein the drying conditions comprise: drying at 20-30 deg.C under normal pressure for 1.5-2.5 hr.
10. The method for preparing a soluble microneedle patch carrying a purified tuberculin protein derivative according to any one of claims 6 to 8, wherein the pressurizing in the step of preparing the base comprises: placing the whole microneedle female die in an environment with the pressure of 0.8-1.2 atmospheric pressure, and pressurizing for 9-11s, wherein the drying conditions comprise: drying at 20-30 deg.C under normal pressure for 5-7 hr.
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| CN108888577B (en) * | 2018-09-11 | 2021-09-14 | 无锡元旭生物技术有限公司 | Soluble hyaluronic acid beauty microneedle patch and preparation method thereof |
| CN111053715A (en) * | 2019-10-22 | 2020-04-24 | 广东雅姿精化有限公司 | High-molecular soluble microneedle and production method thereof |
| CN110947088A (en) * | 2019-12-06 | 2020-04-03 | 广州新济薇娜生物科技有限公司 | Eye-protecting combined patch and preparation method thereof |
| CN112023033B (en) * | 2020-04-29 | 2023-08-25 | 中山大学·深圳 | Two-section micro-needle array patch for simultaneously realizing BCG vaccine inoculation and diagnosis and preparation method thereof |
| CN112274633B (en) * | 2020-09-16 | 2023-11-07 | 广州新济薇娜生物科技有限公司 | Cord Ma Lutai blood glucose-reducing weight-reducing microneedle patch and preparation method and application thereof |
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| CN105581975A (en) * | 2014-11-10 | 2016-05-18 | （株）设模磊 | Micro-needle and micro-needle patch |
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