TW201642834A - Injecting emulsion of protopanaxadiol and preparation method thereof - Google Patents

Abstract

Provided are an injecting emulsion of protopanaxadiol (PPD) and a preparation method thereof. The injecting emulsion includes protopanaxadiol, which is a substance extracted from ginseng, fat phase, emulsifier, and secondary emulsifier. The average particle diameter of the injecting emulsion is about 120 nm or about 110 to 130 nm. Because there is no protopanaxadiol injection at the moment, in comparison with other forms of drug, the injecting emulsion of the present invention is further provided with active features of clear composition, stable quality, excellent anti-tumor effect and easy industrial production. In addition, the injecting emulsion of protopanaxadiol is characterized in being a submicron emulsion, and the submicron emulsion has an average particle diameter of about 120 nm and a Zeta voltage level of -20 to -40 mV.

Description

Original ginseng diol injection emulsion and preparation method thereof

The invention relates to an original ginseng diol injection emulsion and a preparation method thereof, in particular to a microemulsion type containing ginseng extract protopanaxadiol, an oil phase, an emulsifier and a co-emulsifier.

Protopanaxadiol (PPD) is a glycol-type ginsenoside, mainly derived from ginseng, American ginseng, Sanqi and other Araliaceae plants. It is a glycoside of diol group ginsenosides, and is also a microbial glycolysis of diol group ginsenosides. The final product afterwards. Studies have shown that PPD inhibits multiple tumor growth, antidepressant, activates (activates) chloride channels, inhibits sodium channel depolarization initiated (activated), inhibits growth of human embryonic kidney cells HEK-293 cells, and growth of Helicobacter pylori. active. At the same time, PPD has a protective effect on cognitive impairment. ,

PPD is a lipophilic compound with poor solubility in water, low oral bioavailability, and short half-life. It is necessary to study the preparation of protopanaxadiol to improve bioavailability. Mao Jingjing and other researches have prepared PPD dry suspension, which improves drug absorption and is not affected by food. The particles are distributed in the gastrointestinal tract, and the absorption is fast, and the bioavailability is improved (see Mao Jingjing, Zhang Wei, Wang Bing, etc.). 20(S)-Prescription and preliminary quality study of crude panaxadiol dry suspension, Chinese patent medicine, 2013, 34(9): 1680-1684). Jin Shengqi et al. prepared PPD liposomes for intravenous injection in order to achieve targeted delivery and exert anti-tumor effects (see Jin Shengqi, Lei Rongjian, Sun Jingwei: The content and encapsulation efficiency of 20(s)-protopanaxadiol liposome were determined by HPLC, Journal of Nanjing University of Traditional Chinese Medicine, 2009, 25(3): 197-198). Jin Xin et al. prepared the original ginseng lipid cubic liquid crystal nanoparticles of PPD. Lipid cubic liquid crystals formed a gel-like phase cubic structure with special internal structure by polar lipids in the aqueous environment, which has strong adhesion. In order to promote the transmembrane absorption of the drug; the preparation of the square liquid crystal increases the residence time of the drug in the body, and the relative oral bioavailability is 1.66 times that of the raw material drug (see Jin Xin, Zhang Zhenhai, Sun Wei, etc.: the original ginseng diol lipid cube Pharmacokinetics of liquid crystal nanoparticles in rats, Chinese Journal of Traditional Chinese Medicine, 2013, 38(2): 263-268). The emulsion as a carrier can increase the solubility and stability of the poorly soluble drug. The oil phase and the emulsifier can be degraded and absorbed in the body, and the properties are stable. After the intravenous injection, the standard osmotic pressure of the human body is not generated. The invention has been found through patent inquiries and literature searches, and no PPD has been found to be used as an injection emulsion for anti-tumor research.

The technical problem to be solved by the invention is to provide an injectable emulsion for treating tumor by using PPD as a main raw material through the prescription and process screening. The emulsion has stable and controllable quality, simple production process, low cost and broad application prospect.

Another aspect of the invention also provides a process for the preparation of such an emulsion for PPD injection.

Another aspect of the invention also provides the use of such an injectable emulsion for the treatment of tumors.

In order to solve the above technical problem, the present invention is implemented by the following technical solutions:

In one aspect of the invention, there is provided a PPD emulsion for injection, characterized in that the preparation has a PPD of 0.15-0.5%, a vegetable oil of 10-30%, an emulsifier and a co-emulsifier of 3-4%, wherein The proportion of the above raw materials is a percentage by weight.

The invention provides a preparation process of an injection PPD emulsion, comprising: heating and stirring an effective amount of soybean oil for injection, adding an appropriate amount of phospholipid for injection, stirring at a high speed to a clear solution, maintaining a certain temperature and stirring speed, adding an effective amount of PPD The oil phase is obtained after high-speed stirring; the appropriate amount of emulsifier and co-emulsifier are added to the water for injection, and the water phase is obtained by uniformly stirring at a certain temperature; the oil phase is slowly added dropwise to the water phase, and ultrasonic stirring is continued for a certain period of time to obtain colostrum. Adjust the pH value of colostrum to 8.0; homogenize the colostrum high-pressure loop several times, stabilize at room temperature overnight, fill, autoclave.

When the emulsion is used clinically, it is administered by intravenous drip.

In order to test the physical and chemical properties and anti-cancer effects of the microemulsion preparation for PPD injection of the present invention, we have investigated the emulsion as follows:

1. Particle size, distribution and zeta potential: After submicron emulsion is diluted 100 times with water, the particle size of the microemulsion is measured by a laser particle size potentiometer to be 110-130 nm (average particle size is about 120 nm), and (PDI) is 0.1-0.2, Zeta potential is -20~-40mV.

2. Observation of emulsion form: Dilute the appropriate amount of submicron milk to the appropriate concentration, drop the microemulsion on the copper mesh, negatively stain with 2% phosphotungstic acid for 1 min, and observe the spherical droplets uniformly dispersed by transmission electron microscopy. No adhesion.

3. Determination of drug loading and content of the emulsion: After filtering the submicron emulsion through a 0.45um filter, dilute the appropriate multiple and determine the drug content by HPLC to be 400-500mg/100ml.

4. Stability test: The effects of autoclaving, centrifugation, dilution, low temperature (4 °C) and room temperature (20 °C) on the stability of PPD microemulsion were investigated. The results showed that autoclaving, centrifugation and dilution had no effect on the stability of PPD microemulsion, and it was stable for 6 months. Accelerate at room temperature The test (30 ° C, RH 60%) test for 6 months, no significant changes in various indicators.

5. Anti-tumor effect: The killing ability of aPPD on multidrug resistant leukemia cell line P388/ACM cells and human leukemia cell THP-1 cell line was investigated. IC ₅₀ values were 13 and 37.2 uM, respectively. Strong cytotoxicity.

The present invention will be further described in detail with reference to the preferred embodiments of the present invention. It should be noted that these examples are only intended to illustrate the invention and not to limit the scope of the invention.

First Embodiment: The prescription is:

The above-mentioned first embodiment is prepared by stirring soybean oil at 80 ° C and adding 20 g of egg yolk lecithin, dispersing to a clear solution at a high speed, and then adding 5 g of protopanaxadiol at 60 ° C, and uniformly dispersing at a high speed to obtain an oil phase. In addition, 20 ml of glycerin and 15 g of F68 were added to 800 ml of water, and the mixture was stirred at 60 ° C to obtain an aqueous phase; the oil phase was slowly added dropwise to the aqueous phase under high-speed stirring at 60 ° C, and ultrasonic stirring was continued for 5 minutes to a volume of 1000 ml. The colostrum is obtained by circulating the pressure of high pressure homogenization at 700 bar for 10 times.

Second Embodiment: The prescription is:

The preparation method of the above second embodiment: the soybean oil was stirred at 80 ° C and 20 g of egg yolk lecithin was added, and dispersed at a high speed to a clear solution, and then 5 g of protopanaxadiol was added at 60 ° C, and the oil phase was obtained by high-speed dispersion. Further, 20 ml of glycerin and 20 g of F68 were added to 800 ml of water, and the mixture was stirred at 60 ° C to obtain an aqueous phase. The oil phase was slowly added dropwise to the aqueous phase under high-speed stirring at 60 ° C, and ultrasonic stirring was continued for 5 minutes to a volume of 1000 ml. The colostrum is obtained by circulating the pressure of the high pressure homogenized 1000 bar for 5 times.

Third Embodiment: The prescription is:

The preparation method of the above third embodiment: the soybean oil was stirred at 80 ° C and 20 g of egg yolk lecithin was added, and dispersed at a high speed to a clear solution, and then 5 g of protopanaxadiol was added at 60 ° C, and the oil phase was obtained by high-speed dispersion. In addition, 20 ml of glycerin and 10 g of F68 were added to 800 ml of water, and the mixture was uniformly stirred at 60 ° C. Get the water phase. The oil phase was slowly added dropwise to the aqueous phase under high-speed stirring at 60 ° C, and ultrasonic stirring was continued for 5 minutes to a volume of 1000 ml. The colostrum was obtained by high pressure homogenization at 700 bar pressure for 15 times.

Fourth Embodiment: The prescription is:

The preparation method of the above fourth embodiment: the soybean oil was stirred at 80 ° C and 20 g of egg yolk lecithin was added, and dispersed at a high speed to a clear solution, and then 5 g of protopanaxadiol was added at 60 ° C, and the oil phase was obtained by high-speed dispersion. Further, 20 ml of glycerin and 10 g of F68 were added to 800 ml of water, and the mixture was stirred at 60 ° C to obtain an aqueous phase. The oil phase was slowly added dropwise to the aqueous phase under high-speed stirring at 60 ° C, and ultrasonic stirring was continued for 5 minutes to a volume of 1000 ml. The colostrum is obtained by circulating the pressure of 1000 bar under high pressure for 10 times.

Fifth Embodiment: The prescription is:

The preparation method of the above fifth embodiment: stirring soybean oil at 80 ° C and adding 20 g of egg The yellow lecithin was dispersed at a high speed into a clear solution, and then 5 g of protopanaxadiol was added at 60 ° C, and dispersed at a high speed to obtain an oil phase. Further, 20 ml of glycerin and 10 g of F68 were added to 800 ml of water, and the mixture was stirred at 60 ° C to obtain an aqueous phase. The oil phase was slowly added dropwise to the aqueous phase under high-speed stirring at 60 ° C, and ultrasonic stirring was continued for 5 minutes to a volume of 1000 ml. The colostrum was obtained by circulating the pressure of high pressure homogenization at 700 bar for 10 times.

The above is only the preferred embodiments of the present invention, and is intended to be illustrative, and not restrictive, and it is understood by those of ordinary skill in the art that Many changes, modifications, and even equivalents may be made without departing from the scope of the invention.

Claims (5)

An original ginseng diol injection emulsion containing ginseng extract, oil phase, emulsifier and co-emulsifier, characterized in that the original ginseng diol injection emulsion is a submicroemulsion, and the average particle size of the microemulsion It is about 120 nm and the zeta potential is -20--40 mV. The raw ginseng diol injection emulsion according to claim 1, wherein the ginseng extract has a content of proto-glycol diol of 65-85%, and the oil phase is selected from the group consisting of soybean oil and peanut oil, and the emulsifier is selected from the group consisting of injection. With egg yolk lecithin and soy lecithin, the co-emulsifier is selected from the group consisting of F68 for injection and solutol. The ginseng diol injection emulsion according to claim 1 or 2, wherein the ginseng extract emulsion for injection comprises 0.1-0.5% by weight of raw ginseng diol (PPD) and 10-30% by weight of oil phase. Emulsifier and co-emulsifier 3-4%. A method for preparing an original ginseng diol injection emulsion, which is used for preparing an original ginseng diol injection emulsion according to any one of claims 1 to 3, which comprises the following steps: (1) according to the request item 1 to the original ginseng diol injection emulsion according to any one of claim 3, adding an appropriate amount of the drug to the oil phase, heating and dissolving by ultrasonic stirring, adding lecithin for injection, dispersing at a high speed to obtain a clear solution, and then adding effective The original ginseng diol (PPD) is dispersed at high speed to obtain the oil phase; (2) the emulsifier and the co-emulsifier are dissolved in an appropriate amount of water, and ultrasonically heated to form an aqueous phase; (3) under a certain temperature and rotation speed, The aqueous phase is slowly added to the oil phase to prepare a priming emulsion; (4) the colostrum in the above (3) is taken, and the water for injection is added to the prescribed amount, and subjected to high-pressure homogenization cycle emulsification until the particle diameter is 110-130 nm ( The average particle diameter is about 120 nm); (5) The emulsion of the above (4) is obtained by filling and sterilizing. The preparation method according to claim 4, wherein the colostrum is prepared at a temperature of 20-80 ° C, a stirring speed of 100-2000 rpm, an ultrasonic time of 10-60 min, a high-pressure homogenization pressure of 500-2000 bar, and a high-pressure circulation number of 5 -20 times.