CN112494455A

CN112494455A - Colon targeting preparation, preparation method and application

Info

Publication number: CN112494455A
Application number: CN202011433375.9A
Authority: CN
Inventors: 陈振华; 周良良; 胡晓艳
Original assignee: Jiangxi Science and Technology Normal University
Current assignee: Jiangxi Science and Technology Normal University
Priority date: 2020-12-10
Filing date: 2020-12-10
Publication date: 2021-03-16

Abstract

The invention belongs to the technical field of oral colon-targeted drug delivery systems, and discloses a colon-targeted preparation, a preparation method and application thereof, wherein the colon-targeted preparation consists of a solid coating material and a drug; the mass ratio of the solid coating material to the medicine is 90: 1-2: 1. Preparing a solid coating material and a medicament according to a mass ratio of 90: 1-2: 1; grinding and mixing the solid coating material and the medicine. The invention utilizes the surface effect and the special adsorption performance of the ultrafine particles, and directly mixes and grinds the colon-targeted coating material with the average particle size of less than 10 mu m and the drug powder, so that the colon-targeted coating material is adsorbed on the surface of the drug powder, and has the colon-targeted drug release characteristic. The method is simple to operate, does not need a plasticizer, an anti-sticking agent and an organic solvent, reduces the production period and the production cost, enhances the safety in the production process, and has very wide application prospect.

Description

Colon targeting preparation, preparation method and application

Technical Field

The invention belongs to the technical field of oral colon-targeted drug delivery systems, and particularly relates to a colon-targeted preparation, a preparation method and application thereof.

Background

An oral colon targeted drug delivery system is a novel drug delivery mode developed in the later 90 s of the 20 th century, and means that the drug is not released in the upper digestive tract after being orally taken through a drug delivery technology and begins to disintegrate or erode and release after reaching the ileocecal part of a human body, so that the drug can play a local or systemic therapeutic effect in the large intestine of the human body [1 ]. Through oral colon targeted drug delivery, the drug concentration of the colon part can be increased in a targeted manner, and diseases of the colon part, such as colitis, colon cancer and the like, can be treated [2 ]. Meanwhile, the colon targeted drug delivery system can avoid the absorption of the drug in the stomach and small intestine, reduce the toxic and side effect of the drug, or reduce the dosage of the drug. Protein drugs can be safely transported through the stomach and small intestine, and the degradation of the drugs by proteases in the stomach and small intestine is avoided. Therefore, the protein polypeptide drug has great clinical value for treating colon diseases such as colitis, colon cancer and the like and protein polypeptide drugs which are easy to be destroyed in gastrointestinal tracts.

The oral colon targeted drug delivery system has several types, such as pH dependent type, time lag type, enzymatic type, pressure type, etc. [3-4 ]. Currently, pH-dependent studies are used more frequently. The preparation method comprises preparing pH dependent materials such as acrylic resin, plasticizer, antisticking agent, and solvent into coating solution, coating on the surface of granule, tablet, pill, and capsule containing medicine with spray coating equipment, and drying to obtain colon targeting preparation [5-6 ]. In practical application, the coating method has high requirements on the shape, the flowability and the like of the solid medicine, the processes of ingredients and coating are complex, the production period is long, the production cost is high, and the explosion-proof requirement is high when organic solvents such as ethanol and the like are commonly used.

[1] Treford. pharmacy [ M ]. 7 th edition. beijing: the publication of people's health, 2011, 441-.

[2]Bansal V，Malviya R，Malaviya T，et al.Novel prospective in colon specific drug delivery system[J].Polim Med，2014，44(2)：109-118.

[3] The coating formula of the Xubenliang, Chua Yun, Zhang Qi, and the like pH-time lag combined paeonol colon targeting tablet is investigated [ J ] Chinese patent medicine, 2015,37(007):1447-1451.

[4] Zhuxiafang, Yongmei, Tianli, etc. the research progress of the preparation technology of the colon-targeted Chinese medicine [ J ] Chinese patent medicine 2016,38(8):1810-1813.

[5] Yangming, Xie Xinghuang, Mahongyan, etc. preparation of pH-dependent Yuchangning colon targeting tablets and evaluation of in vitro and in vivo release [ J ] Chinese herbal medicine, 2006,37(005):680-684.

[6] Preparation and in-vitro evaluation of a Chongdong, Xuhuinan, Zhang yoga, pH dependence-slow release type mesalazine colon targeting pellet [ J ] Chinese medicine industry journal 2000,31(12): 541) and 544.

Through the above analysis, the problems and defects of the prior art are as follows: the existing coating method has high requirements on the shape, the flowability and the like of solid medicines, the processes of ingredients and coating are complex, the production period is long, the production cost is high, and organic solvents such as ethanol and the like are commonly used, so that the explosion-proof requirement is high.

The difficulty in solving the above problems and defects is: the method for directly grinding and clathrating solid medicines with different properties by using different colon targeting materials selects colon targeting materials with different types and properties according to the property difference of the solid medicines to carry out clathration process research.

The significance of solving the problems and the defects is as follows: the colon-targeted preparation prepared by the method not only meets the characteristics of colon targeting, but also has simple preparation process, low equipment requirement and good preparation formability, does not use organic reagents basically, and greatly reduces the production cost.

Disclosure of Invention

Aiming at the problems in the prior art, the invention provides a colon targeting preparation, a preparation method and application.

The invention is realized by the colon targeting preparation, which consists of a solid coating material and a medicament; the mass ratio of the solid coating material to the medicine is 90: 1-2: 1.

further, the solid coating material is one or a mixture of more than two of chitosan, guar gum, Eudragit L100, acrylic resin II and acrylic resin III.

Furthermore, the medicine is used for treating pathological changes of colon parts such as ulcerative colitis or colon cancer.

Another object of the present invention is to provide a method for preparing the colon-targeted formulation, which comprises the following steps:

firstly, preparing a solid coating material and a medicament according to a mass ratio of 90: 1-2: 1;

and secondly, grinding and mixing the solid coating material and the medicine.

Further, the grinding and mixing are carried out by one or more of a mortar, an ultrafine grinder, a ball mill and a colloid mill.

Further, the medicament is solid powder or a crashed massive solid.

Further, the solid coating material may have an average particle size of 1nm to 10 μm or may be pulverized in advance to have such a particle size range.

The invention also aims to provide a medicament for treating colitis, which is the colon-targeted preparation.

The invention also aims to provide a medicine for treating colon cancer, which is the colon-targeted preparation.

Another object of the present invention is to provide a colon targeted drug delivery system using the colon targeted formulation.

By combining all the technical schemes, the invention has the advantages and positive effects that: the invention is prepared by grinding and mixing the solid coating material and the medicament, wherein the average grain diameter of the solid coating material is within the range of 1 nm-10 mu m, or the solid coating material is crushed into superfine grain diameter in advance. The coating material is mainly one or a mixture of more than two of Eudragit L100, acrylic resin II and acrylic resin III. The medicine is used for treating ulcerative colitis or colon cancer. The invention utilizes the surface effect and the special adsorption performance of the ultrafine particles, and directly mixes and grinds the colon-targeted coating material with the average particle size of less than 10 mu m and the drug powder, so that the colon-targeted coating material is adsorbed on the surface of the drug powder, and has the colon-targeted drug release characteristic.

The invention provides a colon targeting preparation method which directly uses a solid coating material to coat the surface of a medicament without preparing the coating material into a coating solution or a liquid coating material to carry out spray coating and drying.

Drawings

In order to more clearly illustrate the technical solutions of the embodiments of the present application, the drawings needed to be used in the embodiments of the present application will be briefly described below, and it is obvious that the drawings described below are only some embodiments of the present application, and it is obvious for those skilled in the art that other drawings can be obtained from the drawings without creative efforts.

FIG. 1 is a flow chart of a method for preparing a colon targeted formulation provided by an embodiment of the present invention.

FIG. 2 is a scanning electron micrograph of a colon-targeted preparation of Pulsatillae radix of example 1 of the present invention;

in fig. 2: A. the total saponins of the pulsatilla chinensis bunge extract, the guar gum and the pulsatilla chinensis bunge total saponin targeting preparation are prepared; a1, B1, C1 are the amplification results of A, B, C, respectively.

FIG. 3 is a graph of the release rate of the colon-targeted preparation of Pulsatillae radix of example 1 according to the present invention.

Fig. 4 is a graph of the release degree of the colon-targeted baicalin preparation provided by the embodiment of the invention in example 2.

Detailed Description

In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.

Aiming at the problems in the prior art, the invention provides a colon-targeted preparation, a preparation method and application thereof, and the invention is described in detail with reference to the accompanying drawings.

The colon targeting preparation provided by the invention consists of a solid coating material and a medicament; the mass ratio of the solid coating material to the medicine is 90: 1-2: 1.

the solid coating material is one or a mixture of more than two of chitosan, guar gum, Eudragit L100, acrylic resin II and acrylic resin III.

The medicine is used for treating pathological changes of colon parts such as ulcerative colitis or colon cancer.

As shown in fig. 1, the preparation method of the colon targeting preparation provided by the invention comprises the following steps:

s101: according to the mass ratio of 90: 1-2: 1 preparing a solid coating material and a medicament;

s102: grinding and mixing the solid coating material and the medicine.

In the invention, one or more of a mortar, an ultrafine grinder, a ball mill and a colloid mill are adopted for grinding and mixing.

In the present invention, the drug is a solid powder or a pulverized massive solid.

In the present invention, the solid coating material has an average particle diameter in the range of 1nm to 10 μm, or is pulverized in advance to the particle diameter range.

The method for preparing the colon-targeted preparation provided by the invention can be implemented by other steps by those skilled in the art, and the method for preparing the colon-targeted preparation provided by the invention in fig. 1 is only one specific example.

The technical effects of the present invention will be described in detail with reference to specific embodiments.

Example 1:

micronizing guar gum to average particle diameter of about 6 μm, mixing with radix Pulsatillae extract powder at a ratio of 5:1, grinding to uniform color, and observing and recording appearance properties including color, odor, and particle uniformity of the prepared preferable composite micropowder. And (2) carrying out gold spraying on a small amount of samples in an ion sputtering device, observing appearance and appearance characteristics by using a Scanning Electron Microscope (SEM), and filling the rest samples in hollow hard capsules to obtain the colon-targeted preparation.

The capsule is subjected to in vitro release degree test, the test conditions refer to relevant regulations in appendix XA and appendix XD of Chinese pharmacopoeia 10 edition two, and according to a third method (small cup method), the rotating speed is 50r/min, the water temperature is 37 ℃, and dissolution media are respectively as follows: artificial gastric juice (0.1mol/L hydrochloric acid solution) is added in 150mL for 2 h; adding artificial small intestine solution (50 mL of 0.2mol/L sodium phosphate solution at 37 deg.C) into the artificial gastric juice for 4 hr; adding appropriate amount of 2mol/L sodium hydroxide solution and galactosidase into the artificial intestinal juice, and adjusting pH to 7.8 for 4 hr. The release rate results show that the preparation has no release in 2h of artificial gastric juice with pH1.0, less than 10% in 4h of artificial small intestinal juice with pH6.8 and about 90% in 1h of artificial colon juice with pH 7.8. FIG. 2 is a scanning electron micrograph of a colon-targeted preparation of Pulsatillae radix of example 1 of the present invention; in fig. 2: A. the total saponins of the pulsatilla chinensis bunge extract, the guar gum and the pulsatilla chinensis bunge total saponin targeting preparation are prepared; a1, B1, C1 are the amplification results of A, B, C, respectively. FIG. 3 is a graph of the release profile of the colon targeted formulation of Pulsatillae radix of example 1.

Example 2:

the chitosan is subjected to superfine grinding until the average grain diameter is about 100nm, mixed with baicalin powder according to the ratio of 6:1, ground until the color is uniform, and filled into hollow hard capsules to obtain the colon-targeted preparation. Fig. 4 is a graph of the release profile of the baicalin colon targeted formulation of example 2.

The capsules were tested for in vitro release, the test conditions referring to the method in example 1. The release rate results show that the preparation has no release in the artificial gastric juice with the pH value of 1.0 for 2h, the release rate in the artificial small intestinal juice with the pH value of 6.8 for 4h is less than 10 percent, and the release rate in the artificial colon juice with the pH value of 7.8 for 1h is about 95 percent.

Pharmacodynamic experiments below demonstrate the in vivo targeting to the colon and its efficacy of example 1.

(1) Animal grouping and dosing

The SD rats were set up with a normal group, a model group, a bulk drug group of pulsatilla extract, a low dose group, a medium dose group, a high dose group of pulsatilla extract colon targeting agent in example 1, and a positive drug (sulfasalazine) group, each group containing 10 drugs.

Test drug dose: the dosage of the active pharmaceutical ingredient of the Chinese pulsatilla root extract is 200mg kg^-1The low, medium and high dosages of the colon targeting preparation group are respectively 50, 100 and 200 mg.kg^-1(all calculated by the pulsatilla chinensis extract).

Positive drug (sulfasalazine) dose: according to the instruction of medicine use, the clinical initial dose is 0.5g/d, the dosage is increased to 2g/d day by day, and the dosage is 0.5/60X 6-0.05 g kg^-1The daily increment is 0.1 g/kg^-1、0.2g·kg^-1。

(2) Model building and drug delivery

The control group and the model group were allowed to freely feed and drink water, and were fasted for 12h before the model was made without water prohibition. The sulfasalazine dosage group is administered by converting into mouse dosage according to instruction, and the radix Pulsatillae extract raw drug group is administered at 200 mg/kg per day^-1The colon-targeted preparation of the pulsatilla chinensis extract is prepared into a low-dose group, a medium-dose group and a high-dose group, the low-dose group and the medium-dose group are respectively administrated once a day, and 50 mg/kg of the colon-targeted preparation is administrated respectively (measured by the total saponin extract of the pulsatilla chinensis)^-1、100mg·kg^-1、200mg·kg^-1Each group was 20 mL/kg^-1The dosage is administered by gavage, and the blank control group and the model group are filled with equal amount of physiological saline for 7 days. Animals were fasted for 24h without water deprivation 1 day before the end of the experiment, and sacrificed after eye ball removal and blood collection.

(3) Detection index and method

Firstly, macroscopic observation and scoring of colon lesions

Colon tissue was taken after sacrifice, visually observed and scored. The scoring criteria were as follows:

0 minute: the colon is not damaged; 1 minute: mild hyperemia and edema of the colon; and 2, dividing: the colon is obviously edematous, and the intestinal mucosa is rough and granular; and 3, dividing: colonic mucosa ulcer is formed, and the longest diameter of the ulcer is less than or equal to 1 cm; and 4, dividing: forming colonic mucosa ulcer, wherein the longest diameter of the ulcer is between 1 and 2 cm; and 5, dividing: colonic mucosa ulcer is formed, and the longest diameter of the ulcer is more than or equal to 2 cm; 6 min: necrosis of the intestinal wall of the entire colon or death of the animal.

The results of macroscopic scoring of colon lesions in each group of rats are shown in table 1. The results show that the macroscopic grading of the colon of each administration group of rats is reduced compared with that of the model group, wherein the reduction of the low-dose groups of the positive drug (sulfasalazine), the Chinese pulsatilla root extract bulk drug and the colon targeting preparation is obvious.

Table 1 results of macroscopic colon scoring (n ═ 10) for each group of rats

Note: in comparison with the set of models,^**P<0.01

② histopathological observation and scoring

Fixing colon of lesion part with 10% paraformaldehyde, dehydrating with 80%, 95%, and 100% ethanol step by step, clearing xylene, embedding in paraffin, slicing, staining by conventional HE, observing intestinal mucosa injury under light microscope, and grading. The scoring criteria were as follows:

"epithelial cell" score: 0 minute: normal morphology; 1 minute: there is goblet cell loss; and 2, dividing: large loss of goblet cells; and 3, dividing: loss of crypt cells; and 4, dividing: crypt cells are largely lost.

"inflammatory cell infiltration" score: 0 minute: no infiltration; 1 minute: soaking in the basal layer of the recess; and 2, dividing: infiltrating to reach the mucous membrane muscular layer; and 3, dividing: infiltrating into the mucous membrane muscular layer with thickening and obvious edema; and 4, dividing: infiltration reaches the submucosa.

Each rat histopathology total score is the "epithelial cell" score plus the "inflammatory cell infiltration" score.

The results of pathological colon scoring for each group of rats are shown in Table 2. The results show that the pathological score of the colon tissues of rats in the positive drug (sulfasalazine), the pulsatilla root extract crude drug group, the colon targeting preparation middle dose group and the low dose group is remarkably reduced compared with the model group, wherein the reduction degree of the colon targeting preparation low dose group is closer to that of the extract crude drug group.

Table 2 pathological scoring results for rat colon tissue in each group (n ═ 10)

Note: in comparison with the set of models,^*P<0.05, comparing with the model group,^**P<0.01

③ detecting the MPO of the colon tissue

A portion of the colon tissue was taken and weighed accurately, a 10% saline homogenate was prepared using a tissue homogenizer, and a portion was diluted to a 5% homogenate in which the MPO level was measured according to the kit instructions.

The results of determination of MPO content in colon tissue of each group of rats are shown in Table 3. The results show that the MPO content in the colon tissue of the rat in the model group is obviously higher than that in the normal group (P < 0.05). Except for the colon-targeted preparation high-dose group, the MPO content of other groups is reduced compared with that of the model group, wherein the reduction of the pulsatilla chinensis extract bulk drug group and the colon-targeted preparation low-dose group is more obvious.

TABLE 3 MPO content in colon tissue of rats in each group (n ═ 10)

Note: in comparison with the set of models,^*P<0.05

serum TNF-alpha and SOD detection

Blood is taken from the retrobulbar venous plexus of the animal at 3000 r.min^-1Serum is taken out by centrifugation, and TNF-alpha and SOD detection are respectively carried out according to the kit operation instructions.

The results of the TNF-alpha assay in the serum of each group of rats are shown in Table 4. The result shows that the serum TNF-alpha content of the rats in the model group is obviously higher than that in the normal group. The positive drug (sulfasalazine), the Chinese pulsatilla root extract raw drug and the colon targeting preparation have the effect of reducing the content of serum TNF-alpha, wherein the colon targeting preparation low dose and the positive drug (sulfasalazine) are reduced more obviously.

Table 4 serum TNF- α content (n ═ 10) in each group of rats

The results of measuring SOD content in the serum of each group of rats are shown in Table 5. The results show that the SOD content in the serum of the rat in the model group is lower than that in the normal group, the SOD content in the serum of each group except the colon targeting preparation high-dose group in each administration group has a rising trend compared with that in the model group, wherein the rising effect of the positive drug (sulfasalazine), the Chinese pulsatilla root extract raw drug and the colon targeting preparation low-dose is obvious.

Table 5 serum SOD content of each group of rats (n ═ 10)

(5) Results

The pharmacological effects of the colon-targeted preparation of the pulsatilla chinensis extract prepared in the example 1 on rat ulcerative colitis of different doses and the bulk drug of the pulsatilla chinensis extract are compared by using the model through the indexes of animal general condition observation, macroscopic grading of colon lesions, colon histopathology grading, the MPO content of colon tissues, the TNF-alpha content of serum, the SOD content and the like. The results of multiple index tests show that the low dose of the Chinese pulsatilla root extract colon-targeted preparation has the same anti-ulcerative colitis effect as the original Chinese pulsatilla root extract drug with 4 times of the dose, and the colon-targeted preparation prepared in the example 1 can reduce the dosage of the drug and achieve the same pharmacodynamic effect.

The above description is only for the purpose of illustrating the present invention and the appended claims are not to be construed as limiting the scope of the invention, which is intended to cover all modifications, equivalents and improvements that are within the spirit and scope of the invention as defined by the appended claims.

Claims

1. A colon-targeted formulation, which is characterized in that the colon-targeted formulation consists of a solid coating material and a medicament; the mass ratio of the solid coating material to the medicine is 90: 1-2: 1.

2. the colon targeted formulation of claim 1, wherein the solid coating material is one or a mixture of two or more of chitosan, guar gum, Eudragit L100, acrylic resin ii and acrylic resin iii.

3. The colon-targeted formulation of claim 1, wherein the drug is a drug for the treatment of lesions in the colon, such as ulcerative colitis or colon cancer.

4. A method of preparing a colon targeting formulation as claimed in claim 1, wherein the method of preparing the colon targeting formulation comprises the steps of:

step one, according to the mass ratio of 90: 1-2: 1 preparing a solid coating material and a medicament;

and secondly, grinding and mixing the solid coating material and the medicine.

5. The method for preparing a colon targeted drug according to claim 4, wherein the grinding and mixing are performed by one or more of a mortar, a micronizer, a ball mill and a colloid mill.

6. The method of preparing a colon targeted formulation according to claim 4, wherein the drug is a solid powder or a pulverized massive solid.

7. The method for preparing a colon targeted pharmaceutical preparation according to claim 4, wherein the solid coating material has an average particle size of 1nm to 10 μm or is pulverized in advance to the particle size range.

8. A medicine for treating colitis, characterized in that the medicine for treating colitis is the colon-targeted preparation of any one of claims 1 to 3.

9. A medicine for treating colon cancer, which is the colon-targeted preparation of any one of claims 1 to 3.

10. A colon targeted drug delivery system, which uses the colon targeted formulation according to any one of claims 1 to 3.