Preparation method of enteric nanocrystalline composition

Abstract

The invention discloses a preparation method of an enteric nanocrystalline composition, and relates to the technical field of functional foods. The preparation method comprises the following steps: performing superfine grinding on the functional food raw materials to obtain superfine powder; adopting an anti-solvent precipitation method to prepare nanocrystals from the ultrafine powder; adding an absorption enhancer into the obtained nanocrystals, uniformly mixing, and granulating to obtain absorption-promoting composite system particles; adding a CYP3A4 enzyme inhibitor into the absorption-promoting composite system particles to prepare a nanocrystal-promoter-inhibitor compound; and coating the obtained nanocrystal-accelerant-inhibitor compound on a fluidized bed by using a pH sensitive enteric coating material to obtain the enteric nanocrystal composition. According to the invention, the bioavailability of oral functional components is remarkably improved through the synergistic effect of a superfine grinding technology, a nanocrystalline preparation technology, an enteric coating technology, an absorption enhancer technology and a CYP3A4 enzyme inhibitor technology.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of functional food, and particularly relates to a preparation method of an enteric nanocrystal composition. BACKGROUND

[0002] With the increasing attention of people to health, the functional food market has ushered in great development opportunities. However, the traditional functional food has the problems of poor bioavailability and difficult absorption.

[0003] At present, the technologies for improving the bioavailability of functional food mainly include supermicro grinding technology, nanocrystal preparation technology, enteric coating technology, absorption enhancer technology and CYP3A4 enzyme inhibitor technology.

[0004] Among them, the supermicro grinding technology: the supermicro grinding technology can produce micron-level powder (1-100 μm), sub-micron-level powder (0.1-1 μm) and nanometer-level powder (1-100 nm), and is a good material modification technology. The supermicro grinding can change the microstructure of the effective components in the functional food, destroy the original tissue structure, and make the cell wall breaking rate reach more than 95%, which is beneficial to the dissolution of the effective components. However, the effect of simply relying on the supermicro grinding technology to improve the bioavailability is limited.

[0005] Among them, the nanocrystal preparation technology: through methods such as anti-solvent precipitation, the active ingredients form uniformly dispersed nanometer-level crystals, which can significantly increase the solubility and saturation solubility. However, the anti-solvent method usually uses organic solvents, which has the risk of residual solvents, and the nanometer particles are easy to aggregate.

[0006] Among them, the enteric coating technology: the enteric coating is a barrier applied to oral drugs, which can control the absorption position in the digestive tract. The functionality of the enteric coating is largely mediated by the change of the pH value of the environment contacted by the enteric coating product. The selection of the polymer and the thickness of the coating layer are crucial for controlling the pH solubility characteristics of the enteric coating dosage form. However, when the enteric coating technology is applied alone, it is difficult to solve the contradiction with the physical stability of the nanometer particles.

[0007] Among them, the absorption enhancer technology: such as piperine, potassium glycyrrhizinate, chitosan, etc., can enhance drug absorption through mechanisms such as changing the mucosal permeability, inhibiting P-glycoprotein or delaying gastric emptying. Piperine is commonly used as a bioavailability enhancer. Bioavailabity Enhancer is a patented extract containing more than 95% piperine, which is used as a bioavailability enhancer. The technology has several functional patents, including US5,536,506 (use of piperine to improve bioavailability of nutritional compounds) and US5,744,161 (use of piperine as a bioavailability enhancer). Studies have shown that in rat experiments, administration of 20mg / kg piperine can increase the in vivo bioavailability of curcumin by about 154%. Piperine can increase the absorption surface of small intestinal mucosa by changing membrane dynamics, membrane permeability, and inducing the synthesis of proteins related to cytoskeleton function, promote drug absorption, and at the same time act on P-glycoprotein, regulate gastrointestinal secretion, and delay gastric emptying to improve drug bioavailability; however, the effect of piperine alone in improving bioavailability is limited.

[0008] Among them, the CYP3A4 enzyme inhibitor technology: such as curcumin, resveratrol, silymarin, etc. can inhibit the CYP3A4 enzyme system, and resist the first-pass metabolism of active ingredients; however, the effect of using alone is limited.

[0009] In the prior art, the above-mentioned various technologies are mostly used alone or only two of them are combined, and it is difficult to effectively solve the problem of low bioavailability of functional food. Therefore, it is urgent to develop a technical solution that can significantly improve the bioavailability of functional food to solve the problem of low bioavailability of existing functional food SUMMARY

[0010] The purpose of the present application is to provide a preparation method of an enteric nanocrystal composition, which combines ultrafine grinding technology, nanocrystal preparation technology, enteric coating technology, absorption enhancer technology and CYP3A4 enzyme inhibitor technology, realizes the synergistic effect of functional food, and significantly improves its oral bioavailability, solving the problems raised by the prior art.

[0011] To solve the above technical problems, the present application is realized by the following technical scheme:

[0012] The present application is a preparation method of an enteric nanocrystal composition, comprising:

[0013] Step 1, ultrafine grinding the functional food raw materials to D90<5μm to obtain ultrafine powder;

[0014] Step 2, using the anti-solvent precipitation method to prepare nanocrystals with a particle size of 50nm-200nm from the ultrafine powder;

[0015] Step 3, adding an absorption enhancer to the obtained nanocrystals and mixing uniformly to obtain an absorption-promoting composite system granule;

[0016] Step 4, adding CYP3A4 enzyme inhibitor into the absorption promoting complex system particles to obtain nanocrystal-promoter-inhibitor complex;

[0017] Step 5, coating the obtained nanocrystal-promoter-inhibitor complex on the fluidized bed with pH sensitive enteric coating material to obtain enteric nanocrystal composition.

[0018] Further, the step 5 further comprises step 6, which comprises mixing the coated enteric nanocrystal composition with auxiliary materials uniformly, and using a rotary tablet press to obtain functional food enteric nanocrystal tablets.

[0019] Further, the absorption promoter is selected from one or more of piperine, chitosan, dipotassium glycyrrhizinate, medium-chain triglyceride, ginsenoside and β-cyclodextrin.

[0020] Further, the CYP3A4 enzyme inhibitor is selected from one or more of curcumin, silymarin, furanocoumarin, piperide, resveratrol and hesperidin.

[0021] Further, in the step 5, the coating weight gain rate is 3%-5%, and the pH sensitive enteric coating material is selected from hydroxypropyl methylcellulose phthalate, carboxymethyl ethyl cellulose or acrylic acid resin enteric polymer.

[0022] Further, the functional food raw materials in the enteric nanocrystal composition include turmeric, taurine, ginseng, dendrobium officinale, gamma-aminobutyric acid, ganoderma lucidum, pepper, chromium yeast, jujube, pueraria powder, ginkgo, oyster powder, cistanche, rehmannia glutinosa, eucommia ulmoides leaf, vitamin B1 / B2, spina date, celery, cherry, goose peptide and baizhi.

[0023] Further, in the step 2, the anti-solvent is water, and the solvent is ethanol-medium-chain triglyceride mixed solvent.

[0024] Further, the enteric nanocrystal composition is prepared into oral dosage forms including tablets, granules, pellets, capsules, enteric capsules, enteric coated tablets, enteric coated pellets or enteric coated granules.

[0025] The present application has the following beneficial effects:

[0026] The present application improves the bioavailability of oral functional ingredients by the synergistic effect of ultrafine grinding technology, nanocrystal preparation technology, enteric coating technology, absorption enhancer technology and CYP3A4 enzyme inhibitor technology, which can generally reach twice of the traditional process; at the same time, by using suitable coating materials and process parameters, the physical stability of the nanometer particles in the coating process and the subsequent storage process is significantly improved. In addition, the process of the present application can be applied to a variety of functional food active raw materials, and has wide applicability and industrialization potential.

[0027] Of course, implementing any product of the present application does not necessarily require achieving all the advantages described above at the same time. BRIEF DESCRIPTION OF DRAWINGS

[0028] In order to more clearly illustrate the technical solutions of the embodiments of the present application, the drawings needed for the description of the embodiments will be briefly introduced as follows. Obviously, the drawings in the following description are only some embodiments of the present application, and for those skilled in the art, other drawings can also be obtained without creative labor.

[0029] Figure 1 The in vitro dissolution rate comparison chart of the turmeric enteric nanocrystal tablet of the present application and the traditional tablet;

[0030] Figure 2 The pharmacokinetic curve of the turmeric enteric nanocrystal tablet of the present application and the traditional tablet. DETAILED DESCRIPTION

[0031] The technical solutions in the embodiments of the present application will be described clearly and completely below with reference to the drawings in the embodiments of the present application. Obviously, the described embodiments are only some embodiments of the present application, not all embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor are within the scope of protection of the present application.

[0032] In the description of the present application, it should be understood that the terms "opening", "upper", "lower", "thickness", "top", "middle", "length", "inner", "periphery" and the like indicate the orientation or positional relationship, which are only for the convenience of describing the present application and simplifying the description, and do not indicate or imply that the indicated components or elements must have a particular orientation, be constructed and operated in a particular orientation, therefore it cannot be understood as a limitation of the present application.

[0033] Example 1

[0034] The preparation of the turmeric enteric nanocrystal tablet includes:

[0035] Step 1, the turmeric plant raw material is crushed by a super micro-pulverizing vibration machine to D90<5 μm to destroy the cell wall structure, the vibration frequency of the super micro-pulverizing vibration machine is set to 60 Hz, the crushing time is 45 minutes, and the crushing is interrupted every 15 minutes for 5 minutes to prevent overheating and ensure uniform crushing effect, to obtain a super micro-pulverized powder;

[0036] Step 2, the super micro-pulverized powder is dissolved in a 5 mg / mL ethanol-medium-chain triglyceride solution to form a saturated solution, then the obtained solution is dropped into an aqueous solution containing 0.3% povidone K30 at a speed of 2 mL / min, and is treated under ultrasonic assistance for 20 minutes, and nanocrystals are precipitated by the generated supersaturation phenomenon; the average particle size of the obtained nanocrystals is 200±20 nm measured by dynamic light scattering method;

[0037] wherein the volume ratio of medium-chain triglyceride and ethanol is 1:1, the volume ratio of solvent and anti-solvent is 1:6; and the ultrasonic power is 150 W and the frequency is 40 kHz.

[0038] Step 3, 100 mg of nanocrystals, 1 mg of piperine, 3 mg of glycyrrhizic acid dipotassium, and 2 mg of β-cyclodextrin are uniformly mixed, and an appropriate amount of ethanol-water mixed solution (volume ratio 1:1) is added as a binder, and granulation is performed in a high-speed mixing granulator, the granulation conditions are: rotation speed 400 rpm, granulation time 6 minutes; the granulated particles are dried at 40°C to constant weight to obtain a promotion absorption composite system granule;

[0039] Step 4, 1 mg of curcumin, 2 mg of resveratrol, and 4 mg of hesperidin are dissolved in ethanol, and mixed with the promotion absorption composite system granule obtained in step 3 to prepare enteric microspheres by emulsification-solvent evaporation method; the mixture is added to an aqueous solution containing 2% polyvinyl alcohol, and emulsified under high-speed shearing conditions of 8000 rpm for 8 minutes, then the ethanol is evaporated under reduced pressure, and the microspheres are collected by centrifugation and dried at 40°C to constant weight to obtain an enzyme inhibition system integrated enteric microsphere;

[0040] Step 5, the enteric composite microspheres obtained in step 4 are placed in a Wurster type fluidized bed coater, the spraying solution is Eudragit L100-55 enteric coating solution (content about 10%), the inlet air temperature is 55-60°C (the temperature is controlled to protect the activity), the material temperature is 38-42°C, the spraying speed is 200-250 g / min, and the atomization pressure is 0.6-0.8 MPa. Coating to an increase rate of about 4%, to ensure that the product has a dissolution rate of <10% in simulated gastric fluid for 2 hours;

[0041] Step 6, the coated enteric microspheres are mixed with appropriate amount of excipients, and then tabletting is performed by using a rotary tablet press to obtain the curcuma enteric nanocrystal tablet. The tabletting parameters are as follows: pressure 20-30 kN, tablet weight 500 mg / tablet, hardness ≥ 50 N; the excipients include microcrystalline cellulose, sweetener, essence, etc.

[0042] Example 2, the same preparation method as in Example 1 is adopted, and taurine is used to replace curcuma.

[0043] Example 3, the same preparation method as in Example 1 is adopted, and ginseng is used to replace curcuma.

[0044] Example 4, the same preparation method as in Example 1 is adopted, and ganoderma is used to replace curcuma.

[0045] Example 5, the same preparation method as in Example 1 is adopted, and dendrobium officinale is used to replace curcuma.

[0046] Example 6, the same preparation method as in Example 1 is adopted, and gamma-aminobutyric acid is used to replace curcuma.

[0047] Example 7, only the ultrafine grinding technology is used to ultrafine grind the functional food raw materials involved in the above Examples 1-6 to D90 less than 5 μm to obtain functional food tablets.

[0048] Example 8, to verify the dissolution performance of the formula of the present application, the six kinds of functional food enteric nanocrystal tablets prepared in Examples 1-6 and the traditional control tablets prepared in Example 7 are respectively subjected to in-vitro dissolution test, and the test results are shown in Table 1.

[0049] The USPI type rotating basket dissolution tester is used for the test, and the conditions are as follows: rotating basket method, rotating speed 75 rpm, dissolution medium is simulated gastric juice (pH 1.2, first 2 hours) and simulated intestinal juice (pH 6.8, 2-8 hours), dissolution medium volume is 900 mL, temperature is 37±0.5℃. The sample is taken every certain time, and the active ingredient content is determined by using high performance liquid chromatography.

[0050] Table 1: In-vitro dissolution results of enteric nanocrystal tablets and traditional control tablets.

[0051]

[0052] From the above table one can see that the dissolution rate of the preparation of the present application in simulated gastric juice is all <10% in 2 hours, which shows that the enteric coating effectively prevents the release of the active ingredient in the stomach; while in simulated intestinal fluid, the dissolution rate of the preparation of the present application is significantly higher than that of the control group, and the cumulative dissolution rate in 8 hours is all more than 90%, which is much higher than that of the control group (65%-75%). For example, in the simulated intestinal fluid, the cumulative dissolution rate of the curcumin preparation in 8 hours is 95.8%, and that of the control group is only 75.6%, which is increased by about 1.3 times; the dissolution rate of the Dendrobium officinale preparation in 8 hours is 94.1%, and that of the control group is 69.7%, which is increased by 1.4 times. In summary, the present application scheme shows significantly higher dissolution for the six kinds of functional foods.

[0053] Example 9, in order to evaluate the effect of the present application on improving the bioavailability, SD rats were used for bioavailability test.

[0054] Each kind of functional food was used with 6 SD rats (weight 200±20g), which were randomly divided into 2 groups, 3 rats in each group.

[0055] Among them, the experimental group was respectively given the enteric nanocrystal tablets prepared in examples 1-6; the control group was respectively given the functional food tablets prepared by using example 7; and the blood samples were collected at 0, 0.5, 1, 2, 4, 6, 8, 12, 24 hours after administration, and the concentration of the functional food in the plasma was determined by high performance liquid chromatography-mass spectrometry. The pharmacokinetic parameters were calculated, and the results are shown in the following table two. Figure 2

[0056] Table two: comparison of pharmacokinetic parameters of functional food enteric nanocrystal tablets and traditional tablets

[0057]

[0058]

[0059] From table two, it can be seen that the present application scheme significantly improves the oral bioavailability of the functional food.

[0060] Example 10, in order to verify the synergistic penetration effect of absorption enhancer and enzyme inhibitor in the present system, Caco-2 single layer cell model experiment was designed. First, Caco-2 cells were cultured in Transwell chamber for 21 days to differentiate into mature single layer (TEER value 500-1100Ω·cm 2 , and the single layer with TEER≥300Ω·cm 2 was selected for experiment), and curcumin was quantitatively prepared into a concentration of 50μM, and the following test groups were set:

[0061] ​Experimental group 1, CYP3A4 enzyme inhibitors are selected from curcumin, resveratrol and hesperidin in a dose ratio of 1:2:4, and no absorption enhancer is added in experimental group 1;

[0062] Experimental group 2, absorption enhancers are selected from piperine, dipotassium glycyrrhizinate and β-cyclodextrin in a dose ratio of 1:3:2, and no CYP3A4 enzyme inhibitor is added in experimental group 2;

[0063] Experimental group 3, CYP3A4 enzyme inhibitors are selected from curcumin, resveratrol and hesperidin in a dose ratio of 1:2:4, and absorption enhancers are selected from piperine, dipotassium glycyrrhizinate and β-cyclodextrin in a dose ratio of 1:3:2;

[0064] Experimental group 4, the linear additive results of experimental group 1 and experimental group 2.

[0065] That is, experimental group 1 is prepared by the preparation method of example 1 without step 3, experimental group 2 is prepared by the preparation method of example 1 without step 4, experimental group 3 is prepared by the preparation method of example 1, and the following control group is prepared by the preparation method of example 1 without step 3 and step 4.

[0066] After 0.5, 1, 2 and 4 hours of penetration, samples were taken from the receptor pool, the concentration of curcumin in the penetrated liquid was determined, and the apparent permeability coefficient (P_app) was calculated; the results are shown in Table III: the P_app of the control group is about 1.24×10 -6 cm / s; the P_app of group 1 is about 2.86×10 - 6 cm / s; the P_app of group 2 is about 3.12×10 -6 cm / s; the P_app of group 3 is about 7.85×10 -6 cm / s; the theoretical expectation is about 5.98×10 -6 cm / s. The P_app of group 3 (enzyme inhibitor + enhancer combination) is significantly higher than the linear additive effect of group 1 and group 2 (7.85 vs 5.98), indicating that there is a significant synergistic penetration effect (1+1>2 effect).

[0067] Table III, the effect of each group on the penetration rate of curcumin

[0068] Experimental group Apparent permeability coefficient P_app (x 10 -6 cm / s) Relative control group fold Control group 1.24±0.18 1.0 Group 1 (enzyme inhibitor) 2.86±0.31 2.3 fold Group 2 (absorption enhancer) 3.12±0.42 2.5 fold Group 3 (combination) 7.85±0.58 6.3 fold Theoretical expectation 5.98±0.45 4.8 fold

[0069] In addition, the same experiment was conducted on the formulations of the other five functional ingredients (taurine, ginseng, ganoderma, dendrobium officinale, and gamma-aminobutyric acid), and the synergistic effect of the combination of absorption enhancers and CYP3A4 enzyme inhibitors in the other five functional food enteric nanocrystal tablets was tested by the same method, and the results are shown in Table IV.

[0070] Table IV, synergistic effect test results

[0071]

[0072]

[0073] From the analysis of Table 4, the P_app of the combination group is significantly higher than the sum of the effects of the single groups, which verifies the universal synergistic effect of the absorption enhancer and the enzyme inhibitor system in the food system. In summary, the above results prove that the present application realizes the absorption synergistic effect of "1+1>2" by optimizing the formula.

[0074] Example 11, to verify that the present application solves the physical stability contradiction between enteric coating and nanoparticles, a comparative stability test is carried out under accelerated conditions.

[0075] Group A: enteric nanocrystal composition prepared by the technical scheme of the present application; Group B: enteric nanocrystal composition prepared by the traditional process (first prepare nanocrystals, then directly enteric coating);

[0076] The test conditions are 40℃±2℃ and 75%RH±5%RH; the detection time points are 0, 1 and 3 months, and the detection indexes include particle size distribution, aggregation rate, Zeta potential and content, and the results are shown in Table 5.

[0077] Table 5, comparison results of physical stability

[0078] Sample group 3-month aggregation rate (%) Average particle size change (nm) Turmeric inventive technology 4.8 205±15 Taurine inventive technology 4.2 205±15 Ginseng inventive technology 5.1 208±18 Ganoderma inventive technology 4.8 207±16 Dendrobium officinale inventive technology 4.6 206±17 Gamma-aminobutyric acid inventive technology 5.3 209±19 Turmeric traditional technology 22.6 285±35 Taurine traditional technology 21.8 285±35 Ginseng traditional technology 24.3 298±42 Ganoderma traditional technology 22.1 291±38 Dendrobium officinale traditional technology 23.5 287±40 Gamma-aminobutyric acid traditional technology 25.7 302±45

[0079] The results show that the aggregation rate of the nanocrystals of the various functional food enteric nanocrystal tablets prepared by the technical scheme of the present application is less than 5.5% after 3 months of storage, and the average particle size does not increase significantly, while the aggregation rate of the samples prepared by the traditional process is as high as 21.8-25.7%, and the average particle size increases significantly, which proves that the present application effectively solves the physical stability contradiction between enteric coating and nanoparticles.

[0080] In the description of the present specification, the description of the terms "one embodiment", "example", "specific example" and the like means that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present application. In the present specification, the illustrative description of the above terms does not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described can be combined in any one or more embodiments or examples in a suitable manner.

[0081] The preferred embodiments of the application disclosed above are only to facilitate the elucidation of the application. The preferred embodiments do not describe all the details of the application and limit the application to the specific embodiments described. Obviously, many modifications and variations can be made in light of the teachings above. The description is chosen and described in order to best explain the principles of the application and its practical application to thereby enable others skilled in the art to best utilize the application and get the best results from the application. The application is only limited by the claims and their full scope and equivalents.

Claims

1. A method of preparing an enteric nanocrystal composition, characterized by, The preparation method comprises the following steps: Step 1: super-micro grinding the functional food raw material to D90<5 μm to obtain superfine powder; Step 2: preparing nano-crystal with a particle size of 50 nm-200 nm from the superfine powder by using anti-solvent precipitation method; Step 3: adding absorption promoter into the obtained nano-crystal and mixing uniformly to obtain absorption-promoting composite system granules; Step 4: adding CYP3A4 enzyme inhibitor into the absorption-promoting composite system granules to obtain nano-crystal-promoter-inhibitor complex; Step 5: coating the obtained nano-crystal-promoter-inhibitor complex on a fluidized bed with pH-sensitive enteric coating material to obtain enteric nano-crystal composition.

2. A process for the preparation of an enteric nanocrystal composition as claimed in claim 1, wherein, In the step 5, step 6 is further included, which comprises mixing the coated enteric nano-crystal composition with auxiliary materials uniformly, and using a rotary tablet press to obtain functional food enteric nano-crystal tablets.

3. A process for the preparation of an enteric nanocrystal composition as claimed in claim 1, wherein, The absorption promoter is selected from one or more of piperine, chitosan, dipotassium glycyrrhizinate, medium-chain triglyceride, ginsenoside and β-cyclodextrin.

4. A process for the preparation of an enteric nanocrystal composition as claimed in claim 1, wherein, The CYP3A4 enzyme inhibitor is selected from one or more of curcumin, silymarin, furanocoumarin, piperide, resveratrol and hesperidin.

5. A process for the preparation of an enteric nanocrystal composition as claimed in claim 1, wherein, In the step 5, the coating weight gain rate is 3%-5%, and the pH-sensitive enteric coating material is selected from hydroxypropyl methylcellulose phthalate, carboxymethyl ethyl cellulose or acrylic acid resin enteric polymer.

6. A process for the preparation of an enteric nanocrystal composition as claimed in claim 1, wherein, The functional food raw material comprises turmeric, taurine, ginseng, dendrobium officinale, γ-aminobutyric acid, ganoderma lucidum, pepper, chromium yeast, jujube seed, pueraria powder, ginkgo, oyster powder, cistanche, rehmannia glutinosa, eucommia ulmoides leaf, vitamin B1 / B2, spina date seed, celery, cherry, goose peptide and baizhi.

7. A process for the preparation of an enteric nanocrystal composition as claimed in claim 1, wherein, In the step 2, the anti-solvent is water, and the solvent is ethanol-medium-chain triglyceride mixed solvent.

8. A process for the preparation of an enteric nanocrystal composition as claimed in claim 1, wherein, The enteric nano-crystal composition is prepared into oral dosage forms including tablets, granules, pellets, capsules, enteric capsules, enteric coated tablets, enteric coated pellets or enteric coated granules.

9. The preparation method of the enteric nano-crystal composition according to claim 8, wherein the enteric nano-crystal composition is prepared into tablets.

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