Application of pinene and perillyl alcohol in compound ocimum gratissimum preparation

Abstract

The invention relates to application of beta-pinene or perillyl alcohol to inhibition of excitation of intestinal smooth muscle, and preferably relates to application of beta-pinene, perillyl alcohol and eugenol to inhibition of excitation of intestinal smooth muscle.

Description

Technical Field

[0001] The present invention belongs to the field of pharmaceutical technology, and particularly relates to the application of β-pinene or perilla alcohol, preferably in combination with eugenol, in inhibiting the excitation of intestinal smooth muscle. Background Art

[0002] Diarrhea, as a widespread epidemic disease, poses a great threat to human health.

[0003] The Chinese patent ZL03114434.9 of the present inventor discloses a caryopteris oil calcium and aluminum suspension, which includes 0.1%-5% of eugenol, 2%-10% of calcium carbonate, 8%-28% of magnesium trisilicate, 0.2%-7% of aluminum oxide, 0.05%-0.2% of suspending agent, 50%-70% of sucrose, and 0.01%-0.1% of essence. Since it lacks electrolyte components such as sodium, potassium, and iron, the present inventor then replaced the inorganic salts in the original formula with terra flava usta and whitish talc in the Chinese patent ZL200510135621.1; considering that simply supplementing electrolyte solution cannot treat the cause of diarrhea, nor can it significantly shorten the course of diarrhea or reduce the frequency and amount of diarrhea, the present inventor also replaced eugenol with complex caryopteris oil to form a pure traditional Chinese medicine compound caryopteris preparation, which has better diarrhea treatment effect and is especially suitable for acute or subacute diarrhea in children and adults.

[0004] However, in the nearly 20 years since then, the research and development of the compound caryopteris preparation for diarrhea treatment has been stagnant, and even lost the direction of improvement, and there is no improved preparation. However, through long-term research, the present inventor found that compared with terra flava usta and whitish talc, the instability of the caryopteris oil raw material has a greater impact on the efficacy of the compound caryopteris preparation and is not easy to eliminate. Although the present inventor has significantly improved the standard of caryopteris oil (raising the eugenol content from the lower limit of 65% specified in the Chinese Pharmacopoeia to 85%), it is found that this impact is not positively correlated with the eugenol content. In particular, the efficacy of directly using eugenol instead of caryopteris oil is not ideal.

[0005] Through long-term and arduous research, the present inventor found that compared with eugenol, a small amount of β-pinene and perilla alcohol can significantly inhibit the excitation of intestinal smooth muscle. Therefore, it is expected that they are beneficial to improving diarrhea symptoms. Moreover, after they are compounded with eugenol, the effect is also better than or at least not worse than that of caryopteris oil. Therefore, a composition with clear components can be used to replace caryopteris oil to relieve diarrhea and abdominal pain, significantly promoting the modernization of traditional Chinese medicine in the compound caryopteris preparation. Moreover, although the components of caryopteris oil are complex, it also contains a small amount of β-pinene and perilla alcohol. Therefore, the changes to the prescription and preparation method of the (original) compound caryopteris preparation are limited, which can save a large amount of costs. Summary of the Invention

[0006] The problem to be solved by the present invention is to provide an improved compound Eugenia caryophyllata Thunb. preparation for treating diarrhea, especially suitable for acute or subacute diarrhea in children and adults.

[0007] Specifically, in the first aspect, the present invention provides a pharmaceutical composition for treating diarrhea, which contains eugenol, β-pinene, perilla alcohol, terra flava usta and whitish talc. Preferably, the pharmaceutical composition of the first aspect of the present invention consists of eugenol, β-pinene, perilla alcohol, terra flava usta and whitish talc.

[0008] The components in Eugenia caryophyllata Thunb. oil are very complex. The main components are eugenol (CAS No.: 97-53-0), caryophyllene and eucalyptol, etc. There are at least dozens of other detected trace components, and they change with the variety, planting, harvesting and extraction process of Eugenia caryophyllata Thunb., etc. β-pinene (CAS No.: 127-91-3) and perilla alcohol (CAS No.: 536-59-4) have been detected in some Eugenia caryophyllata Thunb. oils respectively, but the content is small and their efficacy in Eugenia caryophyllata Thunb. oil is unclear. Through long-term research, the present inventors have found that low doses of β-pinene and perilla alcohol can significantly inhibit the excitation of intestinal smooth muscle. Especially when they are compounded with eugenol, they can synergistically increase the inhibitory effect, which is even better than, or at least not worse than, Eugenia caryophyllata Thunb. oil.

[0009] Terra flava usta (also known as hearth soil) is mainly composed of silicic acid, aluminum oxide and iron oxide, and also contains a small amount of sodium oxide, potassium oxide, magnesium oxide, calcium oxide, etc. It has a strong effect of neutralizing gastric acid and adsorption, and can be used for chronic diarrhea due to spleen deficiency. Whitish talc (also known as white fu) has a main component of aluminum silicate, has a good effect of protecting intestinal mucosa and adsorbing intestinal toxins, and can be used for chronic diarrhea and dysentery.

[0010] Preferably, in the pharmaceutical composition of the first aspect of the present invention, the weight ratio of eugenol, β-pinene, perilla alcohol, terra flava usta and whitish talc is 0.8-0.95: 0.018-0.022: 0.09-0.11: 2.7-3.3: 7.2-8.8. In a specific embodiment of the present invention, the weight ratio of eugenol, β-pinene, perilla alcohol, terra flava usta and whitish talc is 0.85: 0.02: 0.01: 3: 8. This formula follows a high proportion of eugenol and makes less changes to the prescription of the (original) compound Eugenia caryophyllata Thunb. preparation.

[0011] In the second aspect, the present invention provides a preparation method of the pharmaceutical composition of the first aspect of the present invention, which includes the process of mixing eugenol, β-pinene, perilla alcohol, terra flava usta and whitish talc. In a specific embodiment, the preparation steps are as follows:

[0012] (1) Take terra flava usta and whitish talc and crush them respectively. Preferably, pass them through a 200-mesh sieve, and mix terra flava usta and whitish talc evenly;

[0013] (2) Add β-pinene and perillyl alcohol to eugenol, and then add the mixed oil portion to the powder obtained by mixing the above-mentioned attar of earth and whitish talc, and mix evenly.

[0014] In a third aspect, the present invention provides a pharmaceutical preparation for treating diarrhea, which comprises the pharmaceutical composition of the first aspect of the present invention and pharmaceutically acceptable excipients. The pharmaceutically acceptable excipients used herein refer to fillers, stabilizers, diluents, carriers or other pharmaceutical excipients that are non-toxic for medicinal use and do not interfere with the active ingredient in exerting its medicinal effect. In a specific embodiment, the pharmaceutically acceptable excipients include sodium carboxymethylcellulose, tragacanth gum, simple syrup and water.

[0015] Those skilled in the art can prepare the pharmaceutical composition into various dosage forms according to the treatment purpose and the need of the administration route (such as oral administration), preferably in unit dosage form, such as tablets, pills, capsules (including sustained release or delayed release forms), powders, suspensions, granules, tinctures, syrups, emulsions, suspensions, etc. The pharmaceutical preparation of the present invention can be administered by the administration methods well known to those skilled in the art, such as oral administration, rectal administration, preferably oral administration. Preferably, the pharmaceutical preparation of the third aspect of the present invention is an oral preparation, such as a suspension or a dry suspension, preferably a suspension.

[0016] In a fourth aspect, the present invention provides a method for preparing the pharmaceutical preparation of the third aspect of the present invention, which comprises the process of mixing the pharmaceutical composition of the first aspect of the present invention and pharmaceutically acceptable excipients. In a specific embodiment, the preparation steps are as follows:

[0017] (a) Take tragacanth gum and sodium carboxymethylcellulose, swell them with water, add distilled water and heat to dissolve, then add simple syrup to the tragacanth gum and sodium carboxymethylcellulose solution, boil, filter and cool, preferably cool to 20°C - 25°C;

[0018] (b) Add the pharmaceutical composition of the first aspect of the present invention to the tragacanth gum and sodium carboxymethylcellulose syrup prepared in step (a), grind it evenly with a colloid mill repeatedly, add water and essence, mix evenly, and divide into portions.

[0019] In a fifth aspect, the present invention provides the use of the pharmaceutical composition of the first aspect of the present invention or the pharmaceutical preparation of the third aspect of the present invention in the preparation of a medicament for treating diarrhea, preferably for treating acute or subacute diarrhea in children or adults. In practice, the dosage of this pharmaceutical combination can be determined according to the administration method and the physical condition of the patient, which is obvious to medical staff. Generally, the daily dosage of the oral suspension of the present invention can be 1 - 150 ml. Through clinical research, the optimal dosage of the suspension for adults is 40 ml each time, 3 times a day; for children under 6 months old, it is 3 ml each time, 3 times a day; for children aged 6 months to 1 year old, it is 5 ml each time, 3 times a day; for children aged 1 to 2 years old, it is 10 ml each time, 3 times a day. The optimal dosage of the dry suspension for adults is 5 g each time, 3 times a day; for children aged 1 - 2 years old, it is 3 g each time, and for children under 1 year old, it is 2 g each time, taken in 3 divided doses.

[0020] In a sixth aspect, the present invention provides the use of β-pinene in the preparation of a medicament for inhibiting intestinal smooth muscle excitation; correspondingly, the present invention also provides a method for inhibiting intestinal smooth muscle excitation in vitro, including applying β-pinene to an in vitro specimen of the intestinal smooth muscle.

[0021] In a seventh aspect, the present invention provides the use of perillyl alcohol in the preparation of a medicament for inhibiting intestinal smooth muscle excitation; correspondingly, the present invention also provides a method for inhibiting intestinal smooth muscle excitation in vitro, including applying perillyl alcohol to an in vitro specimen of the intestinal smooth muscle.

[0022] β-pinene or perillyl alcohol can be used alone as an active ingredient to inhibit intestinal smooth muscle excitation, or can be used in combination to inhibit intestinal smooth muscle excitation. Therefore, in an eighth aspect, the present invention provides the use of β-pinene, perillyl alcohol and eugenol in the preparation of a medicament for inhibiting intestinal smooth muscle excitation; correspondingly, the present invention also provides a method for inhibiting intestinal smooth muscle excitation in vitro, including applying β-pinene, perillyl alcohol and eugenol to an in vitro specimen of the intestinal smooth muscle. Preferably, the weight ratio of eugenol, β-pinene and perillyl alcohol is 0.8 - 0.95:0.018 - 0.022:0.09 - 0.11, such as 0.85:0.02:0.1.

[0023] Preferably, in the sixth, seventh or eighth aspect, the intestinal smooth muscle excitation is induced by acetylcholine (such as acetylcholine chloride).

[0024] The beneficial effects of the present invention are as follows. The improved compound Ocimum gratissimum preparation of the present invention maintains and even enhances the significant anti-diarrhea effect of the (original) compound Ocimum gratissimum preparation, helps diarrhea patients recover to a healthy state, is highly effective, has a rapid onset, requires a small dose, has few toxic and side effects, and is convenient for taking, carrying and transporting. It can be stored for more than 5 years without growing bacteria even without adding preservatives. The improved compound Ocimum gratissimum preparation of the present invention significantly promotes the modernization of traditional Chinese medicine of the compound Ocimum gratissimum preparation, with clearer ingredients, which is more convenient for production and quality control. The improved compound Ocimum gratissimum preparation of the present invention also has limited changes to the prescription and preparation method of the (original) compound Ocimum gratissimum preparation, and a large number of original equipment and processes can be used, saving a large amount of costs.

[0025] For the convenience of understanding, the present invention will be described below through specific examples. It should be particularly noted that these descriptions are merely exemplary descriptions and do not constitute a limitation on the scope of the present invention. Based on the discussion in this specification, many modifications of the present invention will be obvious to those skilled in the art. In addition, all references cited in this application are incorporated herein by reference. Detailed implementation manners

[0026] The following examples will specifically describe the present invention. For any details not elaborated, reference can be made to the relevant guidelines of the SFDA, etc., or according to the instructions or manuals provided by the manufacturers of the reagents and instruments specifically used in the experiments.

[0027] Example 1 Research on the effects of Ocimum gratissimum and its components on isolated intestinal smooth muscle

[0028] The small intestine from the duodenum to the upper half of the ileum of a healthy rabbit was excised, the mesentery was removed and cut into intestinal segments 2-3 cm long. After being rinsed clean with Tyrode's solution, it was stored in Tyrode's solution at 4°C to prepare a rabbit intestinal smooth muscle specimen. The constant-temperature smooth muscle bath was filled with Tyrode's solution, the temperature was controlled at 37±0.5°C and a mixture of 95% O2 and 5% CO2 was introduced. Then the two ends of the specimen were fixed to the ventilation hook and the tension transducer respectively, with an initial load of 1 g, and the tension change curve was recorded. After equilibration for about half an hour until the spontaneous contraction and relaxation were stable, the experiment was started.

[0029] During the experiment, except for the blank group, acetylcholine chloride was added to 10 μM in the other groups, and the average tension (F0) within 1 minute after stabilization was recorded. Then, eugenol oil (eugenol content 85.7%), eugenol, β-pinene, perilla alcohol and their combinations were added to each drug group, and the average tension (F1) within 1 minute after stabilization was recorded. The inhibition rate of each drug on the excitation of rabbit intestinal smooth muscle induced by acetylcholine was calculated using the formula (F0 - F1) / F0; the blank group and the acetylcholine chloride group were replaced with an equal volume of Tyrode's solution accordingly; each group was repeated 5 times.

[0030] Table 1 Effects of Drugs on the Excitation of Rabbit Intestinal Smooth Muscle Specimens Induced by Acetylcholine

[0031] Group Final drug concentration (μg / mL) Inhibition rate (%) Blank / 1.32±1.57 Acetylcholine chloride / -0.15±3.72 Ocimum gratissimum oil 50.0 18.90±9.48** Eugenol 42.5 5.14±3.31* β-Pinene 1.0 11.03±9.80* Perilla alcohol 0.5 9.56±2.06** Eugenol + β-Pinene + Perilla alcohol 42.5+1.0+0.5 25.92±12.33**

[0032] Compared with the acetylcholine chloride group, *: p < 0.05; **: p < 0.01.

[0033] As shown in Table 1, eugenol, caryophyllene, β-pinene, and perilla alcohol all had significant inhibitory effects on the excitation of rabbit intestinal smooth muscle induced by acetylcholine. In particular, β-pinene and perilla alcohol could inhibit very significantly at doses much lower than eugenol; although eugenol had a significant inhibitory effect on the excitation of rabbit intestinal smooth muscle, its inhibitory effect was significantly worse than that of eugenol, etc.; the combination of eugenol, β-pinene, and perilla alcohol could not only synergistically inhibit, but also was on average better than eugenol, although there was no significance between the two mixtures, but the former had clear components.

[0034] Example 2 Improved Compound Eugenia aromatica Oral Suspension

[0035] I. Prescription

[0036]

[0037] Add water to make up 100 mL.

[0038] II. Preparation Method

[0039] (1) Weigh 3 g of terra flava usta and 8 g of whitish talc, crush them through a 200-mesh sieve respectively, and mix evenly.

[0040] (2) Drop 0.02 g of β-pinene and 0.01 g of perilla alcohol into 0.85 g of eugenol, gently shake, and then add them to the fine powder of the above-mentioned mixture of terra flava usta and whitish talc, stir and grind evenly to make the liquid fully absorbed.

[0041] (3) Take 0.15 g of tragacanth gum and 1.0 g of sodium carboxymethylcellulose, swell them with 20 ml of water, and then dissolve them by heating with distilled water. Add 45 g of simple syrup to the above-mentioned tragacanth gum and sodium carboxymethylcellulose solution, boil, filter, and cool to obtain.

[0042] (4) Add the powder prepared in step (2) to the tragacanth gum and sodium carboxymethylcellulose syrup prepared in step (3), add 0.05 ml of orange essence, add water to 100 ml, stir with a homogenizer, mix evenly, and immediately subpackage under continuous stirring to obtain the compound eugenol oral suspension.

[0043] III. Efficacy

[0044] The Second Affiliated Hospital of Xi'an Jiaotong University School of Medicine was commissioned to conduct a clinical study on the improved compound eugenol basil suspension and compound eugenol basil suspension (see Chinese Patent ZL200510135621.1) of the present invention for the treatment of acute diarrhea in adults and children. The results are shown in Tables 2 and 3, indicating that the improved compound eugenol basil oral suspension of the present invention has a definite curative effect on the treatment of adult acute diarrhea, and its effect is better than that of the compound eugenol basil suspension, and it can be promoted for use in pediatrics and internal medicine.

[0045] Table 2 Comparison of treatment effects in adults

[0046]

[0047] Table 3 Comparison of treatment effects in children

[0048]

Claims

1. Use of β-pinene in the preparation of a drug for inhibiting intestinal smooth muscle excitation.

2. Use of perillyl alcohol in the preparation of a drug for inhibiting intestinal smooth muscle excitation.

3. Use according to claim 1 or 2, wherein, The said use is the use of β-pinene or perillyl alcohol alone as an active ingredient.

4. Use according to claim 1 or 2, wherein, The said use is the combined use of β-pinene or perillyl alcohol.

5. The application of claim 4, wherein, The said use is the use of β-pinene, perillyl alcohol and eugenol.

6. The application of claim 5, wherein, The weight ratio of eugenol, β-pinene and perillyl alcohol is 0.8 - 0.95:0.018 - 0.022:0.09 - 0.

11.

7. The application of claim 6, wherein, The weight ratio of eugenol, β-pinene and perillyl alcohol is 0.85:0.02:0.

01.

8. Method for inhibiting intestinal smooth muscle excitation in vitro, comprising applying β-pinene, perillyl alcohol and eugenol to an in vitro specimen of the said intestinal smooth muscle.

9. The method of claim 8, wherein, The weight ratio of eugenol, β-pinene and perillyl alcohol is 0.8 - 0.95:0.018 - 0.022:0.09 - 0.

11.

10. The method of claim 9, wherein, The weight ratio of eugenol, β-pinene and perillyl alcohol is 0.85:0.02:0.01.

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