CN114569586B

CN114569586B - Benzenimod enema and preparation method thereof

Info

Publication number: CN114569586B
Application number: CN202210108281.7A
Authority: CN
Inventors: 鲁奕诗; 乔伟光; 智发朝
Original assignee: Individual
Current assignee: Individual
Priority date: 2022-01-28
Filing date: 2022-01-28
Publication date: 2023-12-05
Anticipated expiration: 2042-01-28
Also published as: CN114569586A

Abstract

The invention relates to the technical field of biological medicines, in particular to a bennimod enema and a preparation method thereof. The components of the benomyl enema comprise benomyl and sodium hydroxide; every 10mL of the benomyl enema contains 0.5 mg-5mg of benomyl and 0.1mg-5mg of sodium hydroxide. The benomyl enema has the advantages of small dosage of the benomyl medicine, no organ toxicity, and good safety and curative effect.

Description

Benzenimod enema and preparation method thereof

Technical Field

The invention relates to the technical field of biological medicines, in particular to a bennimod enema and a preparation method thereof.

Background

In the clinical application and popularization process of the medicine, the problem that the medicine is difficult to be absorbed by the organism is unavoidable, and the effectiveness and the safety of the medicine can be seriously influenced. In addition, the local concentration of the medicine is larger or smaller, which can influence the concentration of the medicine penetrating into tissues, and toxic and side effects can be generated. Therefore, the medicine is prepared into a proper dosage form, so that the medicine can be fully absorbed in the body, and the uniformity of the concentration of a medicine system is ensured, so that the medicine effect is ensured, adverse reactions are reduced, and the medicine safety and the medicine quality are ensured.

The bennimod [ (E) -3, 5-dihydroxyl-4-isopropyl stilbene ] is a small molecular compound extracted from a metabolite of nematode symbiotic bacteria, has a direct anti-inflammatory effect, is a new generation of anti-inflammatory drugs, and can be used for treating various major autoimmune diseases such as psoriasis, eczema, ulcerative colitis and various allergic diseases. The anti-psoriasis drug is mainly used for anti-psoriasis treatment in clinical experiments and the treatment effect of the anti-psoriasis drug on inflammatory bowel diseases is explored in laboratories. The medicine is listed as 'national significant science and technology special project' by the science and technology department, and obtains new clinical lot parts of medicines, thereby having good phase II clinical effect. In order to ensure stable efficacy of the benomyl, it is necessary to conduct intensive studies on the dosage form and monitor the efficacy and drug toxicity.

In recent years, the application cases of the formulations of the phenylmod reported in the literature at home and abroad are fewer, some schemes use DMSO to dissolve the phenylmod to prepare external emulsion, and the clinical curative effect of the external emulsion on psoriasis is studied, but in the schemes, the medicament dosage of the phenylmod is larger and the effect is poor. Therefore, on the premise of not increasing the toxicity of organs, the preparation form capable of reducing the dosage of the benzamod medicine is provided, the benzamod medicine can be fully absorbed by organisms, the uniformity of the concentration of a medicine system is ensured, and the medicine has better curative effect and safety.

Disclosure of Invention

Based on the above, the invention provides the benomyl enema and the preparation method thereof, the dosage of the benomyl medicine is small, the benomyl enema has no organ toxicity, and the benomyl enema has better safety and curative effect.

The technical proposal is as follows:

a benomyl enema, the components of which comprise benomyl and sodium hydroxide;

every 10mL of the benomyl enema contains 0.5 mg-5mg of benomyl and 0.1mg-5mg of sodium hydroxide.

In one embodiment, the enema liquid contains 0.8 mg-3mg of benomyl and 0.2mg-3mg of sodium hydroxide per 10mL of the benomyl enema liquid.

In one embodiment, the enema solution of the present invention contains 0.8 to 1.5mg of benomyl and 0.3 to 0.5mg of sodium hydroxide per 10mL of the enema solution of the present invention.

In one embodiment, the benzolimod enema further comprises a buffer and water.

In one embodiment, the buffer is phosphate buffered saline (phosphate buffered saline, PBS).

In one embodiment, the water is used to solubilize the sodium hydroxide and the bennimodide, the remainder being the buffer.

A preparation method of a benomyl enema, which comprises the following steps:

mixing the benjamod with sodium hydroxide to prepare the benjamod enema.

In one embodiment, the process of mixing the benzamod and sodium hydroxide further comprises the step of adding a buffer and water.

In one embodiment, the preparation method of the benomyl enema comprises the following steps:

dissolving the sodium hydroxide and the bennimodide with water, adding the buffer solution to fix the volume, and adding the buffer solution to fix the volume.

In one embodiment, the steps of preparing the benomyl enema are performed under light-shielding conditions.

Compared with the traditional scheme, the invention has the following beneficial effects:

experiments show that if DMSO is used as a solvent, the DMSO is mixed with water to dissolve the benomyl, so that the benomyl is easy to separate out, the DMSO is replaced by a proper amount of sodium hydroxide, the sodium hydroxide is used for dissolving the benomyl, so that enema liquid which is not easy to separate out can be prepared, and covalent-state medicines are changed into ionic states, so that the medicine treatment effect of the benomyl can be improved. The anti-inflammatory effect of the benomyl enema is remarkable, the dosage of the benomyl is small, the benomyl has no visceral toxicity, and the detection of toxic and side effects proves that the preparation has good drug safety. The concentration uniformity of the drug system is good, the drug system can be fully absorbed by organisms, the intestinal inflammation of the experimental mice can be effectively reduced, the liver and kidney functions of the experimental mice can be improved, and the anxiety symptoms of the experimental mice can be improved.

The benomyl enema takes the benomyl, sodium hydroxide and water as raw materials, the raw materials are cheap and easy to obtain, the operation steps are simple and convenient, the application curative effect is good, the dosage of the benomyl medicine is reduced, and the benomyl enema is economical and friendly.

Drawings

FIG. 1 is a graph showing the color comparison of an as-prepared benomyl enema with an enema that is placed under a light source for a long time;

FIG. 2 is a schematic illustration of mass spectrometry identification of enema liquid of example 1;

FIG. 3 is a graph of the weight, disease Activity Index (DAI) and life cycle effects of various examples and control groups on mice;

FIG. 4 is a graph showing the effect of each control on body weight, disease Activity Index (DAI) and survival period of mice;

FIG. 5 is a graph comparing the effect of several groups of examples, controls and comparative examples on colon length, body weight, disease Activity Index (DAI) in colon inflammatory mice;

FIG. 6 is a comparative plot of colon pathology staining of several groups of mice, examples, controls, and comparative examples;

FIG. 7 is a graph showing the comparison of H & E staining of liver and kidney pathology, color Doppler ultrasound, and liver and kidney function tests for several groups of examples, control and comparative examples;

FIG. 8 is a graph comparing the results of open field tests for several groups of examples, control and comparative examples.

Detailed Description

The present invention will be described in further detail with reference to specific examples. The present invention may be embodied in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete.

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 invention belongs. The terminology used herein in the description of the invention is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention.

Terminology

Unless otherwise indicated or contradicted, terms or phrases used herein have the following meanings:

as used herein, the term "and/or," and/or, "and/or" includes any one of two or more of the listed items and also includes any and all combinations of the listed items, including any two or more of the listed items, any or all of the listed items.

In the present invention, "one or more" means any one, any two or more of the listed items. Wherein "several" means any two or more.

In the present invention, the use of "a combination thereof", "any combination thereof", and the like includes all suitable combinations of any two or more of the listed items.

In the present invention, "optional" means optional or not, that is, means any one selected from two parallel schemes of "with" or "without". If multiple "alternatives" occur in a technical solution, if no particular description exists and there is no contradiction or mutual constraint, then each "alternative" is independent.

In the present invention, "preferred" is merely to describe embodiments or examples that are more effective, and it should be understood that they are not intended to limit the scope of the present invention.

In the invention, the technical characteristics described in an open mode comprise a closed technical scheme composed of the listed characteristics and also comprise an open technical scheme comprising the listed characteristics.

In the present invention, the numerical range is referred to, and both ends of the numerical range are included unless otherwise specified.

In the present invention, the content of the components is referred to as mass percent for solid-liquid mixing and solid-solid mixing, and volume percent for liquid-liquid mixing unless otherwise specified.

In the present invention, the term "percent concentration" refers to the final concentration unless otherwise specified. The final concentration refers to the ratio of the additive component in the system after the component is added.

In the present invention, the temperature parameter is not particularly limited, and it is allowed to perform the constant temperature treatment or the treatment within a predetermined temperature range. The constant temperature process allows the temperature to fluctuate within the accuracy of the instrument control.

The bennimod [ (E) -3, 5-dihydroxyl-4-isopropyl stilbene ] is a small molecular compound extracted from a metabolite of nematode symbiotic bacteria, has a direct anti-inflammatory effect, is a new generation of anti-inflammatory drugs, and can be used for treating various major autoimmune diseases such as psoriasis, eczema, concentrated colitis and various allergic diseases. The anti-psoriasis drug is mainly used for anti-psoriasis treatment in clinical experiments and the treatment effect of the anti-psoriasis drug on inflammatory bowel diseases is explored in laboratories. The medicine is listed as 'national significant science and technology special project' by the science and technology department, and obtains new clinical lot parts of medicines, thereby having good phase II clinical effect. In order to ensure stable efficacy of the benomyl, it is necessary to conduct intensive studies on the dosage form and monitor the efficacy and drug toxicity.

In recent years, the application cases of the formulations of the benomyl reported in the literature at home and abroad are fewer, and some schemes use DMSO to dissolve the benomyl to prepare an external emulsion, so that the effectiveness of the external emulsion in clinical treatment of psoriasis is studied. However, in these cases, the amount of the drug of the benomyl is large and the effect is poor. Therefore, on the premise of not increasing the toxicity of organs, the preparation form capable of reducing the dosage of the benzamod medicine is provided, the benzamod medicine can be fully absorbed by organisms, the uniformity of the concentration of a medicine system is ensured, and the medicine has better curative effect and safety.

The technical scheme of the invention is as follows:

a benomyl enema, the components of which comprise benomyl and sodium hydroxide;

The invention takes the finished product of the benomyl as the raw material, and is assisted with a proper amount of sodium hydroxide to prepare enema liquid, so that covalent state medicine is changed into ionic state, and the medicine treatment effect of the benomyl can be improved. The preparation has the advantages of no need of adding other emulsifying agents and gelling agents for mixing, obvious anti-inflammatory effect, less use amount of the bennimodide, no visceral toxicity and good safety of the medicine, and is proved by monitoring toxic and side effects. The concentration uniformity of the drug system is good, the drug system can be fully absorbed by organisms, the intestinal inflammation of the experimental mice can be effectively reduced, the liver and kidney functions of the experimental mice can be improved, and the anxiety symptoms of the experimental mice can be improved.

Alternatively, each 10mL of the benumod enema includes, but is not limited to, 0.5mg, 0.6mg, 0.7mg, 0.8mg, 0.9mg, 1mg, 1.1mg, 1.2mg, 1.3mg, 1.4mg, 1.5mg, 1.6mg, 1.7mg, 1.8mg, 1.9mg, 2mg, 2.1mg, 2.2mg, 2.3mg, 2.4mg, 2.5mg, 2.6mg, 2.7mg, 2.8mg, 2.9mg, 3mg, 3.1mg, 3.2mg, 3.3mg, 3.4mg, 3.5mg, 3.6mg, 3.7mg, 3.8mg, 3.9mg, 4mg, 4.1mg, 4.2mg, 4.3mg, 4.4.5 mg, 4.6mg, 4.7mg, 4.8mg, 4.9mg, 5mg of benumod.

Each 10mL of the bennimod enema comprises 0.1mg, 0.2mg, 0.3mg, 0.4mg, 0.5mg, 0.6mg, 0.7mg, 0.8mg, 0.9mg, 1mg, 1.1mg, 1.2mg, 1.3mg, 1.4mg, 1.5mg, 1.6mg, 1.7mg, 1.8mg, 1.9mg, 2mg, 2.1mg, 2.2mg, 2.3mg, 2.4mg, 2.5mg, 2.6mg, 2.7mg, 2.8mg, 2.9mg, 3mg, 3.1mg, 3.2mg, 3.3mg, 3.4mg, 3.5mg, 3.6mg, 3.7mg, 3.8mg, 3.9mg, 4mg, 4.1mg, 4.2mg, 4.3mg, 4.4mg, 4.5mg, 4.6mg, 4.5mg and 4.5 mg.

Preferably, each 10mL of the benomyl enema liquid contains 0.8 mg-3mg of benomyl and 0.2mg-3mg of sodium hydroxide.

Further preferably, the solution contains 0.8 to 1.5mg of benomyl and 0.3 to 0.5mg of sodium hydroxide per 10mL of the solution.

In one embodiment, the components of the bennimod enema do not contain an emulsifier.

In one embodiment, the components of the bennimod enema do not contain a gelling agent.

In one embodiment, the components of the bennimod enema do not include polymeric gums. It is understood that polymeric gums include, but are not limited to, xanthan gum, carbomer gum, and the like.

In one embodiment, the components of the bennimod enema do not contain a preservative. It will be appreciated that the preservative includes, but is not limited to, sodium benzoate.

In one embodiment, the composition of the benomyl enema further comprises a buffer and water.

Preferably, the buffer is phosphate buffered saline (phosphate buffered saline, PBS).

Preferably, the water is used to dissolve the sodium hydroxide and the bennimodide, the remainder being the buffer.

Alternatively, the pH of the benomyl enema of this invention may be 7.5-8, preferably 7.8, which is close to the pH of the intestinal tract.

It will be appreciated that the above-mentioned benomyl is in the form of powder, and that the benomyl enema of the present invention is a homogeneous brown solution, as shown in fig. 1a, and is stored under a dark condition. Early experiments in this invention found that when the enema was left under a light source for a long time, the homogeneous brown solution was discolored to a colorless transparent solution, as shown in fig. 1b, which not only had no efficacy, but also aggravated intestinal inflammation in experimental mice, presumably related to the possible presence of light instability in the benomyl.

A preparation method of a benomyl enema, which comprises the following steps:

mixing the benjamod with sodium hydroxide to prepare the benjamod enema.

Optionally, the process of mixing the benzamod and sodium hydroxide further comprises the step of adding buffer and water.

The preparation method of the benomyl enema comprises the following steps:

dissolving the sodium hydroxide and the bennimodide with water, and adding the buffer solution to fix the volume.

It will be appreciated that sodium hydroxide solution may be prepared by first dissolving sodium hydroxide in a portion of water. Optionally, the mass fraction of sodium hydroxide solution includes, but is not limited to: 1mg/mL, 5mg/mL, 10mg/mL, 20mg/mL, 30mg/mL, 40mg/mL, 50mg/mL, etc. The bennimodide is then dissolved with sodium hydroxide solution and the remaining water.

Optionally, each step of preparing the benomyl enema is carried out under a dark condition.

It is understood that the method of mixing includes, but is not limited to, resuspension.

It will be appreciated that the sizing may be done in a volumetric flask, EP tube or centrifuge tube.

The following examples are further offered by way of illustration, and the materials used in the following examples are commercially available unless otherwise indicated, and the apparatus used is commercially available unless otherwise indicated, and the processes involved, unless otherwise indicated, are routine selections by those skilled in the art.

In the following specific examples and comparative examples, each step of preparing a bennimod enema was performed under a dark condition.

Example 1

The embodiment provides a benomyl enema and a preparation method thereof, comprising the following steps:

step (1), weighing 1mg of the benzamod powder.

Step (2), weighing 4mg of sodium hydroxide, and dissolving the sodium hydroxide in 100 mu L of sterile deionized water.

And (3) measuring 10 mu L of the sodium hydroxide solution in the step (2), adding the 10 mu L of the sodium hydroxide solution into the powder of the bennimod measured in the step (1), and adding 90 mu L of sterile deionized water for resuspension to enable the powder to be fully dissolved.

And (4) adding PBS (phosphate buffer solution) into the solution in the step (3) to fix the volume to 10mL, repeatedly oscillating for several times, and fully and uniformly mixing to obtain the bennimod enema, and carrying out mass spectrum identification on the enema, wherein as shown in figure 2, the peak of the anion 253.1233 is observed to be the bennimod component.

Examples 2 to 7

Examples 2 to 7 provide a benumod enema and a preparation method thereof, which are basically the same as example 1 except that: the contents of the benjammod and sodium hydroxide in 10mL of the benjammod enema were different, and are shown in Table 1:

TABLE 1

Wherein "-" means not added.

Comparative examples 1 to 6

Comparative example 1 provided 10mL of PBS control.

Comparative examples 2 to 6 provide a benumod enema and a preparation method thereof, which are basically the same as example 1 except that: the composition and amounts of 10mL of the bennimod enema were varied as shown in Table 2.

TABLE 2

Wherein "-" means not added.

Taking comparative example 3 as an example, the preparation steps are as follows:

step (1), weighing 1mg of the benzamod powder.

And (3) weighing 10 mu L of the sodium hydroxide solution in the step (2), adding the 10 mu L of the sodium hydroxide solution into the powder of the benzamod weighed in the step (1), and adding 3mg of xanthan gum, 5mg of sodium benzoate and 90 mu L of sterile deionized water, and re-suspending to fully dissolve the powder.

And (4) adding PBS (phosphate buffer solution) into the solution obtained in the step (3) to fix the volume to 10mL, repeatedly oscillating for several times, and fully and uniformly mixing to obtain the bennimod enema.

Other comparative examples were prepared with reference to comparative example 3.

And (3) constructing an enteritis model: male mice of 6-8 week old C57/BL strain were prepared and randomly divided into 14 groups, with 5 mice per group satisfied. The 3% dextran sodium sulfate (dextran sulphate sodium, DSS) was free-drinked, and the above-described phenylmod enema solutions of examples 1 to 7 and comparative examples 2 to 6 and the PBS control solution of comparative example 1 were simultaneously enemaed on days 3,5, and 7, and a control group was set without using the control solution or the phenylmod enema solution. The specific method for clysis comprises the following steps: the mice were anesthetized with pentobarbital sodium intraperitoneal injection, and after 15min, 100uL enema was infused, and the mice were placed in a head-to-foot position until they were awakened, and the effects on Body Weight (Body Weight%) and Disease Activity Index (DAI) of the mice were recorded daily (a., b.), as well as exhibiting survival (c.) and mortality (d.). The results are shown in fig. 3 and 4.

Wherein, disease activity index (disease active index, DAI) scoring criteria are shown in table 3:

TABLE 3 Table 3

As can be seen from FIG. 3, the enema solutions of examples 1 to 7 were filled, but the addition of the sodium hydroxide and the styrene mod did not directly cause death of the mice, but the addition of sodium hydroxide decreased, for example, in examples 2 and 4, the death of the mice occurred at a later stage, and in example 7, the death of the mice did not occur by adding more sodium hydroxide.

As can be seen from FIG. 4, when a polymer gel such as xanthan gum is added to the phenylmod enema or a preservative such as sodium benzoate is added, the prepared enema can lead to death of mice at a later stage.

In summary, the dosage forms of examples 1 and 7 as the preferred formulation were compared with comparative examples 7 to 8 below, and the effect of the solvent on the efficacy of the above-mentioned bennimod enema was examined.

Comparative examples 7 to 8

Comparative example 7 provides a benomyl enema and its preparation method, which is basically the same as that of example 1 except that: sodium hydroxide was replaced with Olive Oil (OO) as shown in table 4.

Comparative example 8 provides a benomyl enema and its preparation method, which is basically the same as that of example 7 except that: sodium hydroxide was replaced with Olive Oil (OO) as shown in table 4.

TABLE 4 Table 4

	Benzenemomod	Sodium hydroxide	OO
				Example 1	1mg	0.4mg	-
Example 7	1 mg	2mg	-
				Comparative example 7	1mg	-	0.4mg
Comparative example 8	1mg	-	2mg

And (3) constructing an enteritis model: male mice of 6-8 week old C57/BL strain were prepared and randomly divided into 5 groups, satisfying 10 mice per group. The phenylmod enema solutions of examples 1, 7 and comparative examples 7, 8 were simultaneously enemaed with 3% dextran sodium sulfate (dextran sulphate sodium, DSS) free drinking water on days 3,5, and 7, and a control group not using the phenylmod enema solution was set. The specific method for clysis comprises the following steps: the mice were anesthetized with pentobarbital sodium intraperitoneal injection, and after 15min, 100uL enema was filled, and the mice were placed in the head-to-foot position until they were awakened, and the effect on Body Weight (Body Weight%) and Disease Activity Index (DAI) of the mice was recorded daily. On day 8, colon tissue was taken from the mice and colon length was measured, and the results are shown in fig. 5, wherein fig. 5a and 5b show colon length of the colon inflammatory mice; FIG. 5c is a change in mouse body weight; fig. 5d shows the change in Disease Activity Index (DAI). Comparing colon pathology staining of each group of mice to form tissue sections H & E staining, observing inflammatory cell infiltration degree, and observing mucus staining degree by PAS staining, wherein the result is shown in figure 6, and the result of the immunohistochemical staining Ki67 is shown in figure 6a as H & E staining result; FIG. 6b is H & E staining results; FIG. 6c shows PAS staining results. And carrying out detection on liver and kidney functions, heart color Doppler ultrasound and liver and kidney functions of partial mice, wherein the results are shown in figure 7, and figures 7a and 7E are H & E dyeing conditions and heart color illumination detection results; fig. 7b, 7c and 7d are all liver and kidney function test results.

As is clear from fig. 5, 6 and 7, the effect of improving the enema liquid of example 1 and example 7 was good regardless of the colon length, the body weight and the disease activity index. And colon pathology H & E staining showed minimal inflammatory cell infiltration in mice perfused with the enema of example 1 and example 7; PAS staining showed increased colonic mucus secretion in mice infused with the enema of example 1 and example 7; ki67 showed an increased degree of colonic epithelial proliferation in mice infused with the enema of example 1 and example 7, significant anti-inflammatory effect, and no visceral toxicity.

The effect on anxiety behavior of mice was examined by comparison with example 1, comparative example 1 and comparative example 9 below.

Comparative example 9

Comparative example 9 provides a benomyl enema and its preparation method, which is basically the same as that of example 1 except that: sodium hydroxide was not added.

And (3) constructing an enteritis model: male mice of 6-8 week old C57/BL strain were prepared and randomly divided into 5 groups, with 5 mice per group satisfied. The 3% dextran sodium sulfate (dextran sulphate sodium, DSS) was free-drinked, and the above-described phenylmod enema solutions of examples 1, 7 and comparative example 9 and the PBS control solution of comparative example 1 were simultaneously enemaed on days 3,5, and 7, and a control group not using the phenylmod enema solution was set. The specific method for clysis comprises the following steps: the mice were anesthetized by intraperitoneal injection of pentobarbital sodium, and after 15min, 100uL enema was infused, and the mice were placed in the head-low, high position until they were awakened. Open Field Test (OFT) was performed, specifically: the experiment was performed in a quiet environment. On day 8, animals are brought to the experimental place to adapt to the environment for more than 1h, and the mice are placed in the center of the bottom surface in the box, and shooting and timing are performed at the same time. After 5min of observation, the imaging was stopped. The total distance each mouse was subjected to and the residence time in the central area was counted to evaluate anxiety or depression of the mouse emotion. The results are shown in FIG. 8.

As can be seen from fig. 8, the anxiety behavior of the mice of example 1 was improved.

The technical features of the above-described embodiments may be arbitrarily combined, and all possible combinations of the technical features in the above-described embodiments are not described for brevity of description, however, as long as there is no contradiction between the combinations of the technical features, they should be considered as the scope of the description.

The above examples illustrate only a few embodiments of the invention, which are described in detail and are not to be construed as limiting the scope of the invention. It should be noted that it will be apparent to those skilled in the art that several variations and modifications can be made without departing from the spirit of the invention, which are all within the scope of the invention. Accordingly, the scope of protection of the present invention is to be determined by the appended claims.

Claims

1. The benomyl enema for treating inflammatory bowel disease is characterized by comprising the components of benomyl, sodium hydroxide, phosphate buffer solution and water;

each 10mL of the benomyl enema contains 1 mg-2.5 mg of benomyl and 0.4mg-2mg of sodium hydroxide.

2. The benjammod enema for treating inflammatory bowel disease according to claim 1, wherein per 10mL of said benjammod enema contains 1mg of benjammod and 0.4mg of sodium hydroxide.

3. The benjammod enema for treating inflammatory bowel disease according to claim 1, wherein per 10mL of said benjammod enema contains 1mg of benjammod and 1mg of sodium hydroxide.

4. The benjammod enema for treating inflammatory bowel disease according to claim 1, wherein per 10mL of said benjammod enema contains 1mg of benjammod and 2mg of sodium hydroxide.

5. The benjammod enema for treating inflammatory bowel disease according to claim 1, wherein 2.5mg of benjammod and 0.4mg of sodium hydroxide are contained per 10mL of the benjammod enema.

6. The benjammod enema for treating inflammatory bowel disease according to any one of claims 1 to 5, wherein said water is used to dissolve said sodium hydroxide and benjammod.

7. The benjamod enema for treating inflammatory bowel disease according to any one of claims 1 to 5, wherein the pH value of the benjamod enema is 7.5 to 8.

8. A method of preparing a benomyl enema for treating inflammatory bowel disease according to any one of claims 1 to 7, comprising the steps of:

mixing the benomyl, sodium hydroxide, phosphate buffer solution and water to prepare the benomyl enema.

9. A method of preparing a benomyl enema for treating inflammatory bowel disease according to claim 8, comprising the steps of:

dissolving the sodium hydroxide and the benomyl in the water, and adding the phosphate buffer solution to fix the volume.

10. The method for preparing a benjamod enema for treating inflammatory bowel disease according to any one of claims 8 to 9, wherein each step of preparing the benjamod enema is carried out under a dark condition.