CN108158996B

CN108158996B - A fructus Trichosanthis and radix Polygalae saponin spray and its preparation method

Info

Publication number: CN108158996B
Application number: CN201810227312.4A
Authority: CN
Inventors: 董宁霞; 成植温; 吕文文; 高行
Original assignee: Individual
Current assignee: Individual
Priority date: 2018-03-19
Filing date: 2018-03-19
Publication date: 2020-06-02
Anticipated expiration: 2038-03-19
Also published as: CN108158996A

Abstract

The invention belongs to the technical field of pharmaceutical preparations, and particularly relates to a polygalasaponin hexa spray and a preparation method thereof. The invention relates to a polygalasaponin hexaspray, which comprises the following components: polygalasaponin F, pH regulator, preservative, absorption enhancer and distilled water, wherein the pH is 7.0-7.8. The polygalasaponin hexaspray is a brand new product. The formulation of the present invention was obtained by detailed experiments, including screening of absorption enhancers, pH modifiers and preservatives, which were performed in a number of experiments. The polygalasaponin hexa-spraying agent has a reasonable formula, and both a stability test and a cilium toxicity test meet requirements, and the polygalasaponin hexa-spraying agent creatively uses polygalasaponin for preparing a pharmaceutical composition with the effects of relieving cough and eliminating phlegm, and effect experiments prove that the polygalasaponin hexa-spraying agent has excellent effects of relieving cough and eliminating phlegm.

Description

A fructus Trichosanthis and radix Polygalae saponin spray and its preparation method

Technical Field

The invention belongs to the technical field of pharmaceutical preparations, and particularly relates to a polygalasaponin hexa spray and a preparation method thereof.

Background

According to the statistics of the national health departments, nearly 3 hundred million people in China are infected with respiratory diseases every year, wherein cough patients can reach 5 thousands or more. Meanwhile, the more economically developed the region, the higher the incidence of respiratory diseases. Therefore, the cough relieving product has huge consumer groups and wide market prospect. After decades of market cultivation, the market of cough-relieving medicines has grown into a mature market, and the number of cough-relieving products sold in large pharmacies is not less than 40, and the price is high or low. According to the estimation of related departments, 19405 million of urban and rural residents in China have cough symptoms caused by asthma, bronchitis, tuberculosis and cold. According to 12.6 hundred million population, the prevalence rate of cough of urban and rural residents in China is 15.36%. As is known, the existing scale of the cough-relieving and sputum-reducing medicine market in China is about 60 million yuan, the difference between the existing scale and the theoretical market capacity of cough-relieving medicines in China is large, and the retail market of the cough-relieving medicines has a large growth space. Therefore, the development of a new cough-relieving and phlegm-eliminating medicine has very important significance.

Polygalapanin F (PGSF) is an oleanane-type triterpene saponin extracted from polygala japonica of polygala, and has been proved to have the effects of resisting depression, calming, resisting anxiety and hypnosis and possibly developing intelligence-improving effect by enhancing synaptic plasticity. At present, no relevant report on the application of the Chinese medicinal composition in relieving cough and eliminating phlegm exists.

Disclosure of Invention

The invention aims to provide a polygalasaponin hexa spray with the effects of relieving cough and eliminating phlegm and a preparation method thereof.

The technical scheme for realizing the purpose of the invention is as follows:

a radix Polygalae Japonicae saponin caprone spray comprises the following components: polygalasaponin F, pH regulator, preservative, absorption enhancer and distilled water, wherein the pH is 7.0-7.8.

the absorption enhancer can be selected from common absorption enhancers such as sodium dodecyl sulfate, sodium deoxycholate, sodium ethylene diamine tetracetate, HP- β -cyclodextrin, lecithin, sodium salicylate, polyvidone and laurocapram, preferably polyvidone.

The pH adjuster may be selected from commonly used alkaline pH adjusters such as sodium bicarbonate, disodium hydrogen phosphate, sodium hydroxide, calcium hydroxide, potassium hydroxide and meglumine, preferably meglumine.

The preservative may be selected from conventional preservatives, such as chlorobutanol, benzoic acid, sodium benzoate, sorbic acid, potassium sorbate, sorbitol, methylparaben, ethylparaben, propylparaben, butylparaben, benzalkonium chloride and benzalkonium bromide, preferably chlorobutanol.

The content of polygalasaponin F is 100-200 mg/mL^-1。

The content of the absorption enhancer is 2-10%.

The pH is preferably 7.5.

Meanwhile, the invention also provides a preparation method of the polygalasaponin hexa spray, which comprises the following operation steps: accurately weighing herba Polygalae Japonicae saponin, antiseptic and absorption enhancer, adding distilled water, heating to 40 deg.C in water bath oscillator to dissolve completely, adding distilled water, adjusting pH to corresponding range with pH regulator, and bottling into spray bottle.

The polygalasaponin hexa spray is a brand-new product, and polygala japonica is administrated in a spray form, so that the administration is convenient, the administration amount is small, and the polygalasaponin hexa spray is absorbed through mucous membranes and takes effect quickly. The formulation of the present invention was obtained by detailed experiments, including screening of absorption enhancers, pH modifiers and preservatives, which were performed in a number of experiments. The polygalasaponin hexa-spraying agent has a reasonable formula, and both a stability test and a cilium toxicity test meet requirements, and the polygalasaponin hexa-spraying agent creatively uses polygalasaponin for preparing a pharmaceutical composition with the effects of relieving cough and eliminating phlegm, and effect experiments prove that the polygalasaponin hexa-spraying agent has excellent effects of relieving cough and eliminating phlegm.

Detailed Description

The invention will be further illustrated with reference to the following specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. The experimental procedures, in which specific conditions are not noted in the following examples, are generally carried out according to conventional conditions or according to conditions recommended by the manufacturers. All percentages, ratios, proportions, or parts are by weight unless otherwise specified.

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. In addition, any methods and materials similar or equivalent to those described herein can be used in the methods of the present invention. The preferred embodiments and materials described herein are intended to be exemplary only.

Example 1 screening of polygalasaponin hexa-spray absorption enhancer

accurately weighing appropriate amount of polygalasaponin hexa japonica, sodium dodecyl sulfate, sodium ethylene diamine tetracetate, HP- β -cyclodextrin, polyvidone and laurocapram, dissolving with appropriate amount of distilled water, and respectively preparing sodium dodecyl sulfate, sodium ethylene diamine tetracetate and H containing 5%the solution of P- β -cyclodextrin, polyvidone or laurocapram as sample, and the aqueous solution of polygalasaponin hexa without absorption enhancer as blank control, wherein the concentration of polygalasaponin hexa is 100 mg/mL^-1。

Franz diffusion cells are selected as experimental devices, wherein the upper parts of the Franz diffusion cells are dosing cells, and the lower parts of the Franz diffusion cells are receiving cells. The experimental conditions were as follows: receiving liquid: physiological saline; volume of receiving liquid: 8.0 mL; the stirring speed is as follows: 250 rpm; circulating water bath temperature: 37.5 ℃; sampling volume: 0.2 mL; effective diffusion area: 0.8156cm²(ii) a The nasal septum of a newly killed animal is cut off by long-distance surgical scissors, the nasal mucosa is carefully stripped along the nasal septum cartilage by a blade, the blood clot and the mucus are gently cleaned by normal saline, and then the nasal mucosa is put into the normal saline to be stored for later use at 4 ℃. The receiving pool is filled with physiological saline, and the nasal mucosa is fixed between the two diffusion pools, faces the supply pool and is clamped. And starting the automatic stirring and constant-temperature water bath device. After the balance of the circulating water bath at 37 ℃ for 0.5 hour, 100 mu L of the prepared test sample or blank control is dripped on the surface of the mucous membrane by a micro-syringe, and a dosing chamber is sealed by a sealing film to prevent the concentration change caused by the volatilization of the test solution. Samples were taken at 20, 40, 60, 90, 120 minutes and fluid was replenished. After centrifuging the sample at 10000rpm for 10min, taking the supernatant, measuring the content of the drug in the sample, and calculating the cumulative permeation amount per unit area. The cumulative permeation (Qn) is calculated according to formula i. The permeation-increasing Ratio (ER) is used for measuring the absorption-promoting effect of the absorption promoter on the medicament through the nasal mucosa, the calculation method is shown in formula II, and the results are shown in tables 1 and 2.

Qn＝V₀/A(Ⅰ)

ER＝J_E/J₀(Ⅱ)

In the formula: v₀Represents the volume of solution in the receiving well (mL);

a represents an effective diffusion area (cm)²)；

J_EShows the permeation rate (cm & min) of the drug after the application of the absorption enhancer^-1)；

J₀Indicates the intrinsic permeation rate (cm. min) of the drug^-1)

TABLE 1 the cumulative amount of polygalasaponin that permeated through the nasal mucosa after adding 5% of different absorption enhancers

From the results in table 1, it can be seen that the accumulated permeation amount of polygalasaponin according to the present invention through the nasal mucosa is the greatest when povidone is added after the addition of different absorption enhancers.

TABLE 2 permeation rate and permeation increasing factor of different absorption enhancers for polygalasaponin F

Penetration enhancer	J(μg·cm^-2·min^-1)	ER
			Blank control	0.93	-----
Sodium dodecyl sulfate	2.12	2.23
			Ethylenediaminetetraacetic acid sodium salt	2.41	2.16
Povidone	4.21	4.02
			HP- β -cyclodextrin	1.98	1.78
Laurocapram	2.02	2.22

According to the results in table 2, the permeation rate of polygalasaponin is accelerated most obviously when povidone is added, and the permeation increasing rate is the largest.

The experimental results show that the povidone can obviously enhance the in-vitro membrane penetration speed of the polygalasaponin F and shows obviously excellent technical effects compared with other absorption enhancers.

Example 2 preparation of polygalasaponin hexa spray

Polygalasaponin I spray: precisely weighing 10g of polygalasaponin F, 1g of chlorobutanol and 5g of polyvidone, adding 50mL of distilled water, placing on a water bath oscillator, heating to 40 ℃ to completely dissolve the polygalasaponin F, supplementing the distilled water to 100mL, adjusting the pH value to 7.5 by using meglumine, and filling into a spray bottle to obtain the polygalasaponin F with the concentration of 100 mg/mL^-1The spray of (1).

And 2, a polygalasaponin hexa spray II: accurately weighing 12g of polygalasaponin F, 1.2g of chlorobutanol and 6g of polyvidone, adding 50mL of distilled water, placing on a water bath oscillator, heating to 40 ℃ to completely dissolve the polygalasaponin F, supplementing the distilled water to 100mL, adjusting the pH value to 7.5 by using meglumine, and filling into a spray bottle to obtain the polygalasaponin F with the concentration of 120 mg/mL^-1The spray of (1).

Polygalasaponin F spray III: precisely weighing 16g of polygalasaponin F, 1.6g of chlorobutanol and 8g of polyvidone, adding 50mL of distilled water, placing on a water bath oscillator, heating to 40 ℃ to completely dissolve the polygalasaponin F, supplementing the distilled water to 100mL, adjusting the pH value to 7.5 by using meglumine, and filling into a spray bottle to obtain polygalasaponin F with the concentration of 160 mg/mL^-1The spray of (1).

Polygalasaponin F spray IV: precisely weighing 20g of polygalasaponin F, 2g of chlorobutanol and 10g of polyvidone, adding 50mL of distilled water, placing on a water bath oscillator, heating to 40 ℃ to completely dissolve the polygalasaponin F, supplementing the distilled water to 100mL, adjusting the pH value to 7.5 by using meglumine, and filling into a spray bottle to obtain the polygalasaponin F with the concentration of 200 mg/mL^-1The spray of (1).

Example 3 stability study of polygalasaponin F spray

(1) Test for influencing factor

The polygalasaponin hexa-spray I in example 2 is filled and sealed in a 5mL ampoule, and is placed under the conditions of high temperature (40 ℃), illumination (4500 +/-500 Lx) and low temperature (-18 ℃), samples are taken on the 5 th day and the 10 th day respectively, and each index for stability test is detected, and the result is shown in Table 3.

TABLE 3 influence factor test results of GUAZINGJINSANHOU spray

From the above results, it is understood that the polygalasaponin hexa spray of the present invention can exist stably after being left for 10 days under high and low temperature conditions, and only after being left for 10 days under light conditions, the content is reduced and related substances are increased, so that it should be properly stored in a dark place by using a dark device.

(2) Accelerated test

The polygalasaponin hexa-spray I, the polygalasaponin hexa-spray II, the polygalasaponin hexa-spray III and the polygalasaponin hexa-spray IV in the example 2 are respectively placed at 40 ℃ and RH 75%, samples are respectively taken in 0 th, 1 th, 2 th and 3 th months, and various investigation indexes of stability tests are detected, and the results are shown in Table 4.

TABLE 4 acceleration test results of polygalasaponin hexa spray

From the above results, it is clear that the polygalasaponin hexaspray of the present invention can maintain a stable content without increasing related substances when left at 40 ℃ and RH 75% for 6 months.

(3) Long term sample retention at room temperature

The polygalasaponin hexa-spray I, the polygalasaponin hexa-spray II, the polygalasaponin hexa-spray III and the polygalasaponin hexa-spray IV in the example 2 are respectively placed at room temperature, and are sampled in 0 th month and 3 rd month respectively, and all investigation indexes of a stability test are detected. The results are shown in Table 5.

TABLE 5 Long-term room temperature sample retention test results for polygalasaponin hexa-spray

According to the results, the content of the polygalasaponin hexaspray can be kept stable and related substances are not increased after the polygalasaponin hexaspray is placed at room temperature for 6 months.

The stability experiment results show that the polygalasaponin hexa spray has good stability, and the appearance, pH value, content and various indexes of related substances meet the requirements.

Example 4 toxicity test of cilia of Japanese polygalasaponin hexa spray

Accurately weighing appropriate amount of polygalasaponin hexa, chlorobutanol and polyvidone, adding distilled water to dissolve completely, and preparing polygalasaponin hexa with concentration of 100 mg/mL^-1The spray of (4), wherein the pH is adjusted to 7.5 by meglumine. Preparing test samples with povidone concentrations of 1%, 2%, 5% and 10%, respectively, to 100 mg/mL without absorption enhancer^-1The aqueous solution of polygalasaponin was used as a blank control. With physiological salineAs a negative control, the results are shown in Table 6.

Fixing Bufo siccus in supine position, opening oral cavity, dripping 0.5ml of sample, blank control or negative control on palate mucosa, and soaking palate for 0.5 hr. Cutting toad head, and rapidly cutting into pieces of about 3 × 3mm²The palatal mucosa and normal saline are used for cleaning blood clots and impurities, the cilia of the mucosa face upwards and are laid on a glass slide, the glass slide is lightly covered, the cilia movement condition is observed under an optical microscope with the power of 15 multiplied by 10, and then the glass slide is placed in a chromatographic cylinder with a small amount of distilled water and is sealed, so that the water vapor is in a nearly saturated state, and the ambient temperature is 20-25 ℃. Thereafter, the observation was taken at appropriate intervals, and if the cilia continued to move, the cells were returned to the chromatographic cylinder, and the time (LTCM) from the start of administration to the cessation of ciliary movement was recorded. The results are shown in Table 6.

TABLE 6 toxicity test results of cilia of Japanese polygalasaponin hexa spray

Test article	LTCM(min)
		Physiological saline	225.21±14.43
Blank control	12.45±2.46
		1% sample	34.24±5.46
2% sample	72.46±7.43
		5% sample	110.21±4.54
10% sample	182.43±11.23

According to the results, the polygalasaponin hexaspray presents more obvious cilia toxicity when no povidone is added, a certain cilia toxicity is still presented when the addition amount of povidone is 1%, and the toxicity of the polygalasaponin hexaspray to cilia is reduced to an acceptable degree when the addition amount of povidone is more than 2%.

Example 5 cough relieving effect test of polygalasaponin hexa spray

50 healthy guinea pigs with the weight of 400-500 g are selected and randomly divided into five groups of 10 animals. Group 1: blank control group normal saline 0.2 ml/kg; group 2: positive control group ambroxol hydrochloride oral solution 0.2 ml/kg; group 3: the low dose group of the polygalasaponin hexa spray I is 0.5 mL/kg; group 4: 1mL/kg of the dosage group in the polygalasaponin hexa-spraying agent I; group 5: the high-dose group of the polygalasaponin hexa spray I is 2 mL/kg. Injecting 1g/kg of urethane into abdominal cavity of each group of guinea pigs for anesthesia, cutting skin along cervical positive line, and separating vagus nerve; the upper abdominal skin was clamped with an arterial clamp and pulled over a tension transducer to trace the contraction activity of the diaphragm. When the guinea pig coughs, the diaphragm muscle activity is strengthened, and the abdominal wall is dragged, so that the frequency and intensity of coughs can be recorded. The vagus nerve was stimulated with a square wave stimulator at a frequency of 10Hz and a wave width of 10ms, each for 5s, to find the cough threshold. The interval between two stimulations is 5min, the voltage is gradually increased from 1V test, and the minimum voltage value which just causes cough is found, namely the cough causing threshold value of the animal. The test is carried out 3 times continuously, and the average value is taken as the cough inducing threshold value before administration. Then, each test drug is given according to each group, the test is respectively carried out 30 min, 45 min, 60 min, 90 min and 120min after the drug is given, the threshold value before the drug is used for stimulation, and the stimulation voltage is gradually increased or gradually decreased according to the cough-causing condition to find out the cough-causing threshold value, and the result is shown in table 7.

TABLE 7 cough relieving effect test results of polygalasaponin F spray

According to the results, the polygalasaponin hexa spray can obviously increase the cough threshold of guinea pigs, has obvious cough relieving effect, has equivalent effect to ambroxol hydrochloride oral solution when used at medium dose, has better effect than ambroxol hydrochloride oral solution when used at high dose, and has excellent cough relieving effect.

Example 6 test of expectorant effect of polygalasaponin F spray

50 ICR mice with the weight of 20-22 g are selected, the male and female are half of the ICR mice, and the ICR mice are randomly divided into five groups, and each group comprises 10 ICR mice. Group 1: blank control group normal saline 0.2 ml/kg; group 2: positive control group ambroxol hydrochloride oral solution 0.2 ml/kg; group 3: the low dose group of the polygalasaponin hexa spray I is 0.5 mL/kg; group 4: 1mL/kg of the dosage group in the polygalasaponin hexa-spraying agent I; group 5: the high-dose group of the polygalasaponin hexa spray I is 2 mL/kg. The administration is carried out once a day for 7 days continuously, after 30 minutes of the last administration, the mice are intraperitoneally injected with 500mg/KG phenol red (2.5 percent, 0.2mL/10gbw), after 30 minutes of the injection, the cervical vertebrae are taken off to kill the mice, the mice are fixed on an operation plate in an upward direction, the trachea is stripped, the trachea is cut from the lower part of the thyroid cartilage to the first branch, the mice are placed in a test tube containing 2.0mL of physiological saline, and 0.1mL of 1.0mol/L NaOH is added to stabilize the pH value. After 30 minutes, the OD value was measured at 546nm with a UV spectrophotometer, and the results are shown in Table 8.

TABLE 8 test results of phlegm-resolving effect of radix Polygalae Japonicae saponin and radix Aconiti Kusnezoffii Preparata spray

Group of	Animal number (only)	OD value
			Blank control group	10	0.036±0.01
Positive control group	10	0.083±0.02
			Low dose group	10	0.064±0.01
Middle dose group	10	0.090±0.02
			High dose group	10	0.121±0.02

With the increase of the secretion of phenol red, the sputum output of rats also increases. The results show that the effect of the polygalasaponin hexa spray in dosage is equivalent to that of an ambroxol hydrochloride oral solution in increasing the phenol red secretion amount, and the effect of the polygalasaponin hexa spray in dosage is superior to that of the ambroxol hydrochloride oral solution in dosage, so that the effect of the polygalasaponin hexa spray in dosage is prompted to have the effect of reducing phlegm.

The foregoing is merely a preferred embodiment of the invention and is not intended to limit the scope of the invention, which is defined by the claims appended hereto, and any other technical entity or method that is encompassed by the claims as broadly defined herein, or equivalent variations thereof, is contemplated as being encompassed by the claims.

Claims

1. A radix Polygalae Japonicae saponin Heji spray is characterized in that 10g of radix Polygalae Japonicae saponin Heji, 1g of chlorobutanol and 5g of polyvidone are precisely weighed, 50mL of distilled water is added and placed on a water bath oscillator to be heated to 40 ℃ to be completely dissolved, the distilled water is replenished to 100mL, the pH value is adjusted to 7.5 by meglumine, and the mixture is filled into a spray bottle to obtain the radix Polygalae Japonicae saponin Heji with the concentration of 100 mg.mL^-1The spray of (1).

2. A radix Polygalae Japonicae saponin hexane spray is characterized in that 12g of radix Polygalae Japonicae saponin hexane, 1.2g of chlorobutanol and 6g of polyvidone are precisely weighed, 50mL of distilled water is added and placed on a water bath oscillator to be heated to 40 ℃ to be completely dissolved, the distilled water is supplemented to 100mL, the pH value is adjusted to 7.5 by using meglumine, and the mixture is filled into a spray bottle to obtain the radix Polygalae Japonicae saponin hexane with the concentration of 120 mg.mL^-1The spray of (1).

3. A radix Polygalae Japonicae saponin hexane spray is characterized in that 16g of radix Polygalae Japonicae saponin hexane, 1.6g of chlorobutanol and 8g of polyvidone are precisely weighed, 50mL of distilled water is added and placed on a water bath oscillator to be heated to 40 ℃ to be completely dissolved, the distilled water is supplemented to 100mL, the pH value is adjusted to 7.5 by using meglumine, and the mixture is filled into a spray bottle to obtain the radix Polygalae Japonicae saponin hexane with the concentration of 160 mg.mL^-1The spray of (1).

4. A radix Polygalae Japonicae saponin hexane spray is characterized in that 20g of radix Polygalae Japonicae saponin hexane, 2g of chlorobutanol and 10g of polyvidone are precisely weighed, 50mL of distilled water is added and placed on a water bath oscillator to be heated to 40 ℃ to be completely dissolved, the distilled water is replenished to 100mL, the pH value is adjusted to 7.5 by meglumine, and the mixture is filled into a spray bottle to obtain the radix Polygalae Japonicae saponin hexane with the concentration of 200 mg/mL^-1The spray of (1).