CN109453106B

CN109453106B - Trehalose suppository and preparation method and application thereof

Info

Publication number: CN109453106B
Application number: CN201910020975.3A
Authority: CN
Inventors: 宋淑亮; 曹琪; 邢茂辰; 吉爱国
Original assignee: Shandong University
Current assignee: Shandong University
Priority date: 2019-01-09
Filing date: 2019-01-09
Publication date: 2021-06-25
Anticipated expiration: 2039-01-09
Also published as: CN109453106A

Abstract

The invention provides a trehalose suppository and a preparation method and application thereof, belonging to the technical field of pharmaceutical preparations. The trehalose suppository comprises a pharmaceutical active ingredient trehalose; and pharmaceutically acceptable auxiliary materials. The invention provides the trehalose suppository for preventing or treating atherosclerosis for the first time by matching trehalose with other matrix components, the medicament of the suppository has high dissolution rate, stable property and no irritation, effectively solves the problems of medicament inactivation caused by oral administration of trehalose, poor compliance of injection administration and the like, and has good practical application value and industrial application prospect.

Description

Trehalose suppository and preparation method and application thereof

Technical Field

The invention belongs to the technical field of pharmaceutical preparations, and particularly relates to a trehalose suppository as well as a preparation method and application thereof.

Background

The information in this background section is only for enhancement of understanding of the general background of the invention and is not necessarily to be construed as an admission or any form of suggestion that this information forms the prior art that is already known to a person of ordinary skill in the art.

Atherosclerosis is a chronic inflammatory disease, is the main pathological basis of ischemic cardiovascular and cerebrovascular diseases such as coronary heart disease, cerebrovascular disease, thromboembolic disease and the like, and seriously harms human health. The pathogenesis of the traditional Chinese medicine is complex, and the traditional Chinese medicine is lack of effective prevention and treatment medicines due to the combined action of a plurality of factors.

Trehalose is a non-reducing sugar composed of two glucose molecules with 1, 1-glycosidic bonds, widely exists in many low-grade ferns, algae, fungi, yeasts and insects, and is a safe and reliable natural sugar. The prior research shows that the trehalose can reduce atherosclerotic plaques, thereby playing a role in preventing and treating atherosclerosis.

However, oral administration of trehalose is inactivated by degradation of the gastrointestinal tract by intestinal microorganisms, resulting in ineffective oral absorption and achieving a better effect of treating atherosclerosis only by injection. For people who prevent or treat atherosclerosis for a long time, long-term administration is needed, and the administration mode through injection is not only very inconvenient, but also poor in patient compliance, and seriously affects the treatment effect.

Disclosure of Invention

Aiming at the defects in the prior art, the invention provides a trehalose suppository for preventing or treating atherosclerosis and a preparation method and application thereof.

The invention aims to provide a trehalose suppository.

The invention also aims to provide a preparation method of the trehalose suppository.

The invention also aims to provide application of the trehalose suppository.

In order to achieve the purpose, the invention relates to the following technical scheme:

in a first aspect of the present invention, there is provided a trehalose suppository comprising a pharmaceutically active ingredient trehalose; and pharmaceutically acceptable auxiliary materials. The trehalose suppository can be directly acted on the cavity for administration, preferably rectum, has high drug dissolution, no irritation, large drug-loading rate, less administration times, simple and convenient use, low price, economy and practicability; can effectively avoid the degradation of intestinal microorganisms in oral administration.

The pharmaceutically acceptable auxiliary materials are selected from water-soluble matrixes and antibiotics which are not absorbed by intestinal tracts;

wherein the water-soluble matrix comprises glycerol, gelatin or polyethylene glycol; the antibiotic not absorbed by intestinal tract comprises aminoglycoside antibiotics, and is further preferably gentamicin or tobramycin;

further, the trehalose suppository is prepared from the following raw materials in parts by weight:

5-40 parts of trehalose, 4-10 parts of gelatin, 4-20 parts of glycerol and 0.05-0.1 part of tobramycin or gentamicin;

or the like, or, alternatively,

5-40 parts of trehalose, 02-5 parts of PEG 40002, 01-3 parts of PEG 600and 0.05-0.1 part of tobramycin or gentamicin;

furthermore, the trehalose suppository is prepared from the following raw materials in parts by weight:

28 parts of trehalose, 6 parts of gelatin, 6 parts of glycerol and 0.08 part of gentamicin;

or the like, or, alternatively,

20 parts of trehalose, 40003 parts of PEG, 60002 parts of PEG and 0.08 part of gentamicin.

In a second aspect of the present invention, there is provided a method for preparing the trehalose suppository, comprising:

s1, adding water into the water-soluble matrix according to the prescription amount, and heating the water-soluble matrix in a water bath until the water-soluble matrix is completely melted;

s2, adding the pretreated trehalose into the step S1; after being stirred uniformly, the antibiotics which are not absorbed by the intestinal tract in the prescription amount are added, and the mixture is continuously stirred to be uniform;

s3, filling the mould, standing the mould in a low-temperature environment, and opening the mould to obtain the trehalose suppository.

Wherein, in the step S1,

taking a water-soluble matrix according to the prescription amount, adding 0.4-2 times of water, and heating the water-soluble matrix in a water bath until the water-soluble matrix is completely melted at 65-75 ℃ (preferably 70 ℃).

In the step S2, in the above step,

the trehalose pretreatment method comprises the following steps: trehalose is crushed and sieved by a 100-mesh sieve, and the particle size of the trehalose not only influences the dissolution rate of the finally prepared suppository, but also obviously influences the stability of the suppository.

In the step S3, the temperature is controlled to be 0-4 ℃ (preferably 4 ℃), and the standing time is 0.2-1 h (preferably 0.5 h). The uniform mixture of each component of the suppository can be completely solidified by controlling the standing temperature and the standing time, and the suppository is easy to demould.

In a third aspect of the invention, the trehalose suppository is used for preparing a medicament for preventing or treating atherosclerosis.

The invention has the beneficial effects that:

the trehalose suppository for preventing or treating atherosclerosis provided by the invention is reasonable in compatibility, convenient to apply, fast in effect and short in treatment course, and trehalose can be absorbed by the body; by optimizing the selection and the proportion of the raw materials, the stability and the dissolution rate of the medicine are effectively improved, the bioavailability of the suppository is further improved, the problems of medicine inactivation caused by oral trehalose and poor compliance of injection administration and the like are effectively avoided, and the suppository has good practical application value and industrial application prospect.

Detailed Description

It should be noted that the following detailed description is exemplary and is intended to provide further explanation of the disclosure. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs.

It is noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of example embodiments according to the present application. As used herein, the singular forms "a", "an" and "the" are intended to include the plural forms as well, and it should be understood that when the terms "comprises" and/or "comprising" are used in this specification, they specify the presence of stated features, steps, operations, devices, components, and/or combinations thereof, unless the context clearly indicates otherwise.

As mentioned above, trehalose can reduce atherosclerotic plaques, thereby acting as a prophylactic and therapeutic agent for atherosclerosis. However, oral administration of trehalose is inactivated by degradation by intestinal microorganisms, resulting in ineffective oral absorption and achieving a good therapeutic effect on atherosclerosis only by injection. For people who prevent or treat atherosclerosis for a long time, long-term administration is needed, and the administration mode through injection is not only very inconvenient, but also poor in patient compliance, and seriously affects the treatment effect.

In view of the above, in one embodiment of the present invention, there is provided a trehalose suppository comprising a pharmaceutically active ingredient trehalose; and pharmaceutically acceptable auxiliary materials.

wherein the water-soluble matrix comprises glycerol, gelatin or polyethylene glycol; by reasonably optimizing and selecting the water-soluble matrix material, the finally prepared suppository has strong toughness, elasticity and difficult breakage, and can be slowly dissolved in body fluid in a cavity to release the medicine. It can be mixed with trehalose to be used as sustained release suppository for slowly releasing trehalose.

The antibiotic not absorbed by intestinal tract comprises aminoglycoside antibiotics, and is further preferably gentamicin or tobramycin; in order to avoid the loss of activity of trehalose due to degradation by intestinal microorganisms, the antibiotic which is not absorbed by the intestinal tract is preferably selected and added by comprehensively considering the stability, the dissolution rate and the like of the suppository, so that the activity of the intestinal microorganisms can be inhibited, and the trehalose is prevented from being decomposed into monosaccharide by the intestinal microorganisms to lose the drug effect; meanwhile, the suppository finally prepared keeps better stability and dissolution rate.

In another embodiment of the invention, the trehalose suppository is prepared from the following raw materials in parts by weight:

In another embodiment of the present invention, the tobramycin and gentamicin are both 4 ten thousand units/mL.

In another embodiment of the present invention, there is provided a method for preparing the trehalose suppository, comprising:

s1, taking the water-soluble matrix according to the prescription amount, adding 0.4-2 times of water, and heating in a water bath until the water-soluble matrix is completely melted at 65-75 ℃ (preferably 70 ℃);

s3, filling a mold, placing the mold in a low-temperature environment of 0-4 ℃ for 0.2-1 h, and opening the mold to obtain the trehalose suppository.

In another embodiment of the present invention, in step S2,

In the step S3, the temperature is controlled to be 4 ℃, and the standing time is 0.5 h. The uniform mixture of the suppository components can be completely solidified by adopting the temperature and the time, and the suppository is easy to be demoulded.

In another embodiment of the present invention, there is provided a use of the trehalose suppository as described above for the preparation of a medicament for the prevention or treatment of atherosclerosis.

To further illustrate the present invention, the following detailed description of the invention is given by way of specific examples, which are intended to be purely exemplary and are not to be construed as limiting the invention. Wherein, the tobramycin and gentamicin used in each example have the specification of 4 ten thousand units/mL.

Example 1

The formula is as follows:

taking 4g of gelatin, adding 1 time of water by weight, and soaking until the gelatin is completely swelled; putting the gelatin in a water bath kettle, heating at a constant temperature of 70 ℃ to melt; adding 14g of glycerol, and uniformly stirring to uniformly mix the glycerol and the gelatin; weighing 10g of trehalose, crushing, sieving with a 100-mesh sieve, adding the trehalose into a glycerol gelatin solution, and uniformly stirring; adding 0.08g of tobramycin, and uniformly stirring; and (3) stopping heating after the water is evaporated to the residual 2g, filling the mould, standing at the constant temperature of 4 ℃ for 30 minutes, and opening the mould to obtain the trehalose suppository.

Example 2

The formula is as follows:

adding 2g of water into 40003 g of PEG and 60002 g of PEG, placing in a water bath kettle, heating at the constant temperature of 70 ℃, and stirring and uniformly mixing; weighing 20g of trehalose, crushing, sieving with a 100-mesh sieve, dissolving in water, adding the matrix, and uniformly stirring; adding 0.08g of gentamicin, and uniformly stirring; and (3) stopping heating after the water is evaporated to the residual 2g, filling the mould, standing at the constant temperature of 4 ℃ for 30 minutes, and opening the mould to obtain the trehalose suppository.

Example 3

The formula is as follows:

taking 6g of gelatin, adding 2 times of water by weight, and soaking until the gelatin is completely swelled; putting the gelatin in a water bath kettle, heating at a constant temperature of 70 ℃ to melt; adding 6g of glycerol, and uniformly stirring to uniformly mix the glycerol and the gelatin; weighing 28g of trehalose, crushing, sieving with a 100-mesh sieve, dissolving in water, adding the dissolved trehalose into a glycerol gelatin solution, and uniformly stirring; adding 0.08g of gentamicin, and uniformly stirring; and (3) stopping heating after the water is evaporated to the residual 4g, filling the mould, standing at the constant temperature of 4 ℃ for 30 minutes, and opening the mould to obtain the trehalose suppository.

Example 4

In vitro release experiments were performed on the trehalose suppositories prepared in example 3, and the trehalose content was measured by the phenol-sulfuric acid method. Weighing 0.14g of trehalose, 0.03g of gelatin and 0.03g of glycerol, and metering to a volumetric flask of 1000 mL. 0, 0.2mL, 0.4mL, 0.6mL, 0.8mL, and 1.0mL of the standard solutions were pipetted into a 10mL stoppered tube, and the volume was adjusted to 1.0mL with distilled water. To the sample solution, 1.0mL of 5% phenol solution was added, followed by rapid addition of 5.0mL of concentrated sulfuric acid. Mix quickly and then place the tube in a 30 ℃ water bath for 20 min. An appropriate amount of the reaction solution was taken and absorbance was measured at 490 nm. And drawing a standard curve y which is 9.7837x +0.0185 by taking the mass concentration of the trehalose as an abscissa and the light absorption value as an ordinate.

Taking 6 suppositories, putting the suppositories into a dissolution cup, taking 1000mL of double distilled water as a dissolution medium, timing sampling 0.2mL (simultaneously immediately supplementing an equivalent isothermal fresh medium) at 37 ℃, adding distilled water to supplement 1mL, adding 5% phenol and concentrated sulfuric acid in sequence, developing, measuring a light absorption value, and calculating the total sugar content by a standard linear equation in the same step of preparing a standard curve. As can be seen from Table 1, the trehalose suppositories were fully thawed and completely released within 35 minutes.

TABLE 1 in vitro drug Release percentage

Example 5

In vivo absorption experiments were performed on the trehalose suppositories prepared according to the present invention (example 3) using SD male rats as subjects. Healthy SD rats were randomly grouped into groups of 5 rats each weighing 200 + -20 g, and fasted for 24h before administration, during which time water was freely available. Trehalose suppositories were rectally administered at a dose of 100mg/kg, anal clamped in a suitable manner to prevent leakage, and rats were fixed for 2-4h after administration and fasted for 6 h. 0.3mL of blood is respectively taken from orbital veins of rats at 0min, 15min, 30min, 60min, 90min, 120min, 180min and 240min, and added into an EP tube containing EDTA anticoagulant. Centrifuging at 3000r/min for 15min, collecting supernatant to obtain plasma sample, and storing the plasma sample in a refrigerator at-40 deg.C for detection.

Enzyme-linked immunoassay is adopted, and a trehalose kit is used for detection, so that the content of trehalose in blood is calculated. As can be seen from Table 2, trehalose clearly peaked at 58.4. mu.g/mL at 60 minutes after administration of the trehalose suppository.

TABLE 2 plasma drug concentrations at various times after dosing in rats

Experimental example 6

The preparation process was the same as in examples 1-3, except that trehalose was crushed and sieved through a 60 mesh sieve to obtain trehalose suppositories, which were used as comparative examples 1-3 for stability effect experiments, respectively.

The trehalose suppositories of examples 1 to 3 and the control group were placed in a constant temperature environment (25. + -. 2 ℃) for 6 months at a relative humidity of 65. + -. 5%, and samples were taken at the end of the test period of 1, 3 and 6 months, and the properties of the suppositories, the amount of trehalose (indicated amount%) and the melting time were measured as specified in the Chinese pharmacopoeia 2015, and the test results are shown in Table 3.

TABLE 3 suppository accelerated test results

As can be seen from table 3, the trehalose suppository prepared in example 3 is placed in a constant temperature and high humidity environment for 6 months, the color of the suppository is unchanged, the texture is uniform and fine, the surface of the suppository is smooth and has no cracking phenomenon, the amount of trehalose is not obviously changed, the melting time limit is not obviously prolonged, and the stability is good, while the adverse observation of the control groups 1-3 shows that the surface of the suppository is rough and has serious cracking phenomenon after being placed for 3 months, so that the test cannot be continued, and the melting time limit is obviously prolonged by changing the particle size of trehalose from the viewpoint of the melting time limit; the trehalose suppository of the invention has better stability.

It should be noted that the above examples are only used to illustrate the technical solutions of the present invention and not to limit them. Although the present invention has been described in detail with reference to the examples given, those skilled in the art can modify the technical solution of the present invention as needed or equivalent substitutions without departing from the spirit and scope of the technical solution of the present invention.

Claims

1. A trehalose suppository for preventing or treating atherosclerosis, which is characterized by comprising the following raw materials:

5-40 parts of trehalose, 4-10 parts of gelatin, 4-20 parts of glycerol and 0.05-0.1 part of tobramycin or gentamicin; or the like, or, alternatively,

the preparation method of the trehalose suppository comprises the following steps:

s1, adding water into the water-soluble matrix according to the prescription amount, and heating the water-soluble matrix in a water bath until the water-soluble matrix is completely dissolved;

s3, filling a mold, standing in a low-temperature environment, and opening the mold to obtain the trehalose suppository;

in the step S1, in the above step,

controlling the adding amount of water to be 0.4-2 times of the amount of the water-soluble matrix, and heating the water-soluble matrix in a water bath to 65-75 ℃;

in the step S2, in the above step,

the trehalose pretreatment method comprises the following steps: pulverizing trehalose, and sieving with 100 mesh sieve;

in the step S3, in the above step,

controlling the temperature to be 0-4 ℃; standing for 0.2-1 h;

wherein the water-soluble matrix is glycerol and gelatin, or PEG 4000 and PEG 6000; the antibiotic not absorbed by the intestinal tract is tobramycin or gentamicin.

2. The trehalose suppository of claim 1, wherein the trehalose suppository comprises the following raw materials:

28 parts of trehalose, 6 parts of gelatin, 6 parts of glycerol and 0.08 part of gentamicin.

3. The trehalose suppository of claim 1, wherein the trehalose suppository comprises the following raw materials:

4. The trehalose suppository of claim 1, wherein in step S1, the suppository is heated to 70 ℃ in a water bath.

5. The trehalose suppository of claim 1, wherein, in step S3, the temperature is controlled to 4 ℃; the standing time is 0.5 h.