A composition for relieving nausea and vomiting

Abstract

The present invention relates to the field of medicine and food technology, and relates to a composition for relieving nausea and vomiting, and specifically to a composition obtained by mixing, heating and beating with ginseng and ginger as main raw materials, and its application in the preparation of a drug for relieving / treating nausea and vomiting. In the composition, the mass ratio of ginseng to ginger is 1:10-1:60, and the mass ratio of the mixed raw materials of ginseng and ginger to water is 1:10-1:40. The composition comprises total ginsenosides and total gingerols, wherein the total ginsenoside content is 15-55%, the total gingerol content is 2.5-9%, and the rare ginsenoside content accounts for more than 60% of the total ginsenoside content of total ginseng. The composition is prepared by the following method: mixing ginseng with fresh ginger, soaking in water, mixing and beating at 60-100°C for 30-75min, that is. The composition of the present invention can significantly relieve nausea and vomiting, and can be used to treat nausea and vomiting caused by various reasons.

Description

Technical Field

[0001] The present invention relates to the technical field of medicine and food, and in particular to a composition for relieving nausea and vomiting, and in particular to a composition obtained by mixing, heating and beating ginseng and ginger as main raw materials, and application of the composition in preparing medicine or food for relieving / treating nausea and vomiting. Background Art

[0002] Nausea and vomiting are common symptoms. They are normal physiological reactions of the human body to reduce damage to the digestive tract. They often cause adverse reactions such as dehydration, electrolyte imbalance, and malnutrition, which affect the patient's treatment compliance. Nausea and vomiting can be divided into many types, such as chemotherapy vomiting, postoperative nausea and vomiting, and vomiting during pregnancy. Chemotherapy-related nausea and vomiting (CINV) is the most common adverse reaction, which significantly reduces the quality of life and compliance of cancer patients. In recent years, despite the continuous updating and improvement of drug antiemetic programs and non-drug interventions, 30-50% of CINV are still not effectively controlled. Motion sickness, also known as motion sickness, is a disease caused by a series of vestibular and autonomic nervous system reactions such as dizziness, vertigo, nausea, vomiting, pale complexion, and loss of appetite due to the inability of people to adapt to acceleration, visual and deep sensory stimulation. It brings great pain to people and even seriously affects their normal travel and work.

[0003] Ginger and ginseng are typical representatives of medicine and food, and have a history of medicinal use in my country for thousands of years. Ginseng is the dried root and rhizome of Panax ginseng CAMey., a plant of the Araliaceae family. It has multiple functions such as strengthening the body, tonifying the spleen and benefiting the lungs. In traditional medicine, ginseng is used to treat vomiting and fatigue caused by chemotherapy or radiotherapy. Ginger is the fresh rhizome of Zingiber officinale Rose., a plant of the Zingiberaceae family. It is pungent and slightly warm in nature. It belongs to the lung, spleen, and stomach meridians. It has the functions of dispelling cold, warming the middle and stopping vomiting, resolving phlegm and relieving cough, and detoxifying fish and crabs. It is known as the "holy medicine for vomiting". As a natural antiemetic Chinese medicine, ginger not only has a significant inhibitory effect on nausea and vomiting, but is also safe, cheap, convenient, and easy to obtain. It can avoid the adverse reactions of chemical antiemetics and has good health care effects. In recent years, modern pharmacological research has made important progress in the antiemetic effect and mechanism of ginger, confirming that ginger alone has a good antiemetic effect. At present, there are no reports on the use of ginger in combination with other drugs for antiemetics.

[0004] Food and medicine products have broad prospects and advantages of effectiveness, safety and economy in the prevention and treatment of CINV, and can reduce the economic burden of patients to a certain extent. Summary of the invention

[0005] In order to overcome the defects of the prior art, the technical problem solved by the present invention is to provide a composition for relieving nausea and vomiting, wherein the composition is prepared by mixing, heating and beating ginseng and ginger as raw materials.

[0006] To achieve the above object, the technical solution adopted by the present invention is:

[0007] The invention provides a composition for relieving nausea and vomiting. The composition is prepared by taking ginseng and ginger as raw materials and mixing and beating the raw materials after adding water.

[0008] The mass ratio of ginseng to ginger is 1:10-1:60, preferably 1:30-1:60;

[0009] The mass ratio of the mixed raw material of ginseng and ginger to water is 1:10-1:40.

[0010] The composition of the invention comprises total ginsenosides and total gingerols, wherein the total ginsenosides are the sum of prototype ginsenosides and rare ginsenosides.

[0011] The total ginsenoside content is 15-55%, the total gingerol content is 2.5-9%, and the rare ginsenoside content accounts for more than 60% of the total ginsenoside content.

[0012] Furthermore, the total ginsenoside content is 30-55%, the total gingerol content is 3.5-9%, and the rare ginsenoside content accounts for more than 70% of the total ginsenoside content.

[0013] Furthermore, the total ginsenoside content is 30-55%, the total gingerol content is 5-9%, and the rare ginsenoside content accounts for more than 75% of the total ginsenoside content.

[0014] in,

[0015] The total ginsenoside content is the percentage of total ginsenosides to the total mass of ginseng.

[0016] The total gingerol content is the percentage of total gingerol to the total weight of ginger.

[0017] The total ginsenosides contain the following components: ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, ginsenoside Rc, ginsenoside Rg2, ginsenoside Rb2, ginsenoside Rh1, ginsenoside Rd, ginsenoside Rg6, ginsenoside F4, ginsenoside Rk3, ginsenoside Rh4, ginsenoside S-Rg3, ginsenoside R-Rg3, ginsenoside Rk1, ginsenoside Rg5;

[0018] The weight percentage composition of each component is as follows: ginsenoside Rg1 0.51-1.99%, ginsenoside Re 0.78-3.63%, ginsenoside Rb1 2.16-3.56%, ginsenoside Rc 0.61-2.70%, ginsenoside Rg2 0.73-1.87%, ginsenoside Rb2 0.34-1.68%, ginsenoside Rh1 0.01-0.29%, ginsenoside Rd 0.21-1.63%, ginsenoside Rg6 0.01-2.11%, ginsenoside F4 0.05-2.27%, ginsenoside Rk3 0.01-1.19%, ginsenoside Rh4 5.54-33.25%, ginsenoside S-Rg3 0.07-1.75%, ginsenoside R-Rg3 0.01-0.19%, ginsenoside Rk1 0.08-0.69%, ginsenoside Rg5 0.02-2.12%.

[0019] Preferably,

[0020] The weight percentage composition of each component is as follows: ginsenoside Rg1 0.51-1.99%, ginsenoside Re 0.78-2.72%, ginsenoside Rb1 2.24-3.56%, ginsenoside Rc 0.61-2.70%, ginsenoside Rg2 0.73-1.43%, ginsenoside Rb2 0.34-1.68%, ginsenoside Rh1 0.01-0.29%, ginsenoside Rd 0.21-1.63%, ginsenoside Rg6 0.01-1.13%, ginsenoside F4 0.05-1.12%, ginsenoside Rk3 0.38-1.19%, ginsenoside Rh4 19.53-33.25%, ginsenoside S-Rg3 0.11-1.75%, ginsenoside R-Rg3 0.01-0.04%, ginsenoside Rk1 0.23-0.69%, ginsenoside Rg5 0.17-2.12%.

[0021] More preferably,

[0022] The weight percentage composition of each component is as follows: ginsenoside Rg1 1.89-1.99%, ginsenoside Re 1.71-1.81%, ginsenoside Rb1 3.53-3.56%, ginsenoside Rc 2.10-2.70%, ginsenoside Rg2 1.37-1.39%, ginsenoside Rb2 1.38-1.68%, ginsenoside Rh1 0.09-0.29%, ginsenoside Rd 0.83-1.00%, ginsenoside Rg6 0.78-0.98%, ginsenoside F4 0.05-0.25%, ginsenoside Rk3 1.17-1.19%, ginsenoside Rh430.25-33.25%, ginsenoside S-Rg3 1.65-1.75%, ginsenoside R-Rg3 0.01-0.03%, ginsenoside Rk1 0.59-0.69%, ginsenoside Rg5 0.17-0.27%.

[0023] The total gingerol contains the following components: 6-shogaol, 6-gingerol, 8-gingerol, and 10-gingerol;

[0024] The weight percentage of each component is as follows: 0.98-3.66% of 6-shogaol, 0.98-2.17% of 6-gingerol, 0.06-1.53% of 8-gingerol and 0.16-1.54% of 10-gingerol.

[0025] Preferably,

[0026] The weight percentage of each component is as follows: 1.23-3.66% of 6-shogaol, 1.25-2.17% of 6-gingerol, 0.72-1.32% of 8-gingerol, and 0.34-1.54% of 10-gingerol.

[0027] Preferably,

[0028] The weight percentage of each component is as follows: 1.66-3.66% of 6-shogaol, 2.07-2.17% of 6-gingerol, 0.81-1.03% of 8-gingerol, and 0.53-1.54% of 10-gingerol.

[0029] The present invention provides a method for preparing the above composition, comprising the following steps:

[0030] The ginseng and fresh ginger are mixed, soaked in water, mixed and beaten at 60-100° C. for 30-75 minutes to obtain a pharmaceutical composition using ginseng and ginger as raw materials.

[0031] Preferably,

[0032] The ginseng and fresh ginger are mixed, soaked in water, mixed and beaten at 90-100° C. for 45-60 minutes to obtain a pharmaceutical composition using ginseng and ginger as raw materials.

[0033] The mass ratio of the mixed raw material of ginseng and ginger to water is 1:10-1:40.

[0034] The invention can effectively increase the content of total ginsenosides, especially rare ginsenosides, in ginseng by mixing ginseng with ginger and then heating and beating the mixture, and simultaneously increase the content of total gingerols in ginger, thereby improving the effect of treating nausea and vomiting.

[0035] The rare ginsenosides include: Rg2, Rb2, Rh1, Rg6, F4, Rk3, Rh4, S-Rg3, R-Rg3, Rk1, and Rg5.

[0036] Experiments have shown that the content of ginsenoside Rh4 in the composition is significantly increased, and ginsenoside Rh4 accounts for 20-70% of the total ginsenosides, preferably 50-70%, and more preferably 60-70%.

[0037] The present invention provides a good vomiting relief effect of the above-mentioned composition. The composition has a relieving effect on a rat pica model caused by cisplatin for a certain period of time. Studies have shown that the composition has a significant relieving effect on the pica reaction caused by cisplatin, and its effect is better than that of ginseng or ginger alone or the addition of the two.

[0038] The composition of the present invention can be prepared into clinically acceptable preparations with pharmaceutically acceptable carriers or excipients.

[0039] The present invention provides application of the composition or its preparation in preparing a medicine for relieving / treating nausea and vomiting.

[0040] The nausea and vomiting include nausea and vomiting caused by chemotherapy drugs, nausea and vomiting caused by radiotherapy, postoperative vomiting, vomiting during pregnancy, motion sickness, etc.

[0041] The chemotherapy drug is one or more of cisplatin, paclitaxel, etoposide, teniposide, daunorubicin, doxorubicin, idarubicin, cyclophosphamide, mitoxantrone, aclarubicin, homoharringtonine, methotrexate, vincristine, and vindesine.

[0042] The nausea and vomiting described in the present invention may also refer to vomiting caused by gastrointestinal damage, inflammatory diseases, and anorexia.

[0043] Furthermore, the composition or preparation thereof of the present invention can be used to relieve nausea and vomiting caused by cisplatin.

[0044] The present invention provides application of the composition or its preparation in preparing food for assisting in protecting gastric mucosa.

[0045] The present invention further provides application of ginger in increasing the content of rare saponin in ginseng.

[0046] The invention also provides application of ginseng in increasing the total gingerol content of ginger.

[0047] The food includes health food.

[0048] Compared with the prior art, the present invention has the following beneficial effects:

[0049] The invention prepares the composition by mixing ginseng and ginger and beating them to increase the content of effective components, wherein the content of rare ginsenosides is increased by more than 40%, the content of Rh4 is increased by more than 70%, and the content of total gingerol is increased by more than 400%.

[0050] The invention can effectively improve the efficiency of converting the prototype saponins in ginseng into rare saponins by mixing ginseng with ginger and then heating and beating. The content of rare saponins in ginseng is increased, and the content of gingerol in ginger is also increased. That is, ginseng and ginger are mixed and then beaten, and under the interaction of the two, not only the content of rare saponins in ginseng is increased, but also the content of gingerol in ginger is increased, so that the combination of the two plays a better antiemetic effect.

[0051] After a lot of research and experiments, the present invention finally obtained the composition with the best drug ratio and preparation conditions of the present invention, which is an innovative composition that matches the ratio of multiple receptor targets, and has been proven to have excellent therapeutic effects on side effects such as vomiting caused by cancer chemotherapy. The composition is not only highly effective and low in toxicity, but also economical, easy to take, and easy to apply and promote. BRIEF DESCRIPTION OF THE DRAWINGS

[0052] Ginseng and ginger combination (GG), normal group (Control), model group (Model), ginseng group (GY), ginger group (JY), ginseng and ginger combination low-dose group (GGL) and ginseng and ginger combination high-dose group (GGH).

[0053] Figure 1 The effects of GG on body weight, kaolin and normal feed intake of pica rats in Example 6.

[0054] A is the effect of GG on the body weight of pica rats;

[0055] B is the effect of GG on kaolin intake in pica rats;

[0056] C is the effect of the composition on the normal feed intake of pica rats.

[0057] Figure 2 This is the effect of GG in Example 7 on 5-HT in the serum and medulla oblongata of pica rats.

[0058] A is the medullary content in serum;

[0059] B is the 5-HT content in the medulla oblongata.

[0060] Figure 3 This is the effect of GG in Example 8 on the expression of 5-HT, MAOA, TPH2 and TPH1 proteins in the ileum and medulla oblongata of pica rats.

[0061] A is the protein band expression diagram; B is 5-HT; C is TPH2; D is MAO-A; and E is TPH1.

[0062] Figure 4 This is the histopathological observation of GG on gastrointestinal damage caused by cisplatin in Example 9 and the relief of gastric antrum and intestinal damage.

[0063] Figure 5 This is the effect of GG in Example 10 on gastric emptying and small intestinal motility in mice chemotherapeutic with cisplatin.

[0064] A is the gastric emptying rate; B is the small intestinal propulsion rate.

[0065] Figure 6 This is the effect of ginseng puree and the composition in Example 11 on IEC-6 cell viability.

[0066] Figure 7 This is the effect of ginger puree and the composition in Example 12 on IEC-6 cell viability.

[0067] Figure 8 The effect of gingerol and shogaol in GG in Example 14 on the viability of IEC-6 cells.

[0068] Fig. 9 The pharmacodynamic effect of GG on motion sickness was verified by detecting behavioral changes in rats in each group through the heterophilic kaolin experiment in Example 15.

[0069] All data in the above figures are expressed as mean±SD, and “#” indicates a significant difference compared with the normal group (P<0.05). # P<0.05, ## P<0.01, ### P<0.001; "*" indicates significant difference compared with the model group (P<0.05). * P<0.05, ** P<0.01, *** P<0.001. DETAILED DESCRIPTION

[0070] The present invention will be further described below by specific examples. These examples should be understood to be only used to illustrate the present invention and not to limit the scope of protection of the present invention. Those skilled in the art can make various changes or modifications to the present invention based on the content of the present invention, and these equivalent changes and modifications also fall within the scope defined by the claims of the present invention.

[0071] Unless otherwise specified, other materials, reagents, etc. used in the following embodiments and effect examples can be obtained from commercial sources.

[0072] In the following examples, the types and contents of ginsenosides, gingerols and shogaols in the ginseng and ginger composition GG were detected using HPLC technology, and the specific operation was as follows:

[0073] Carry out the test under the following chromatographic conditions, obtain the peak area of ​​the reference substance, then measure the peak area of ​​the sample corresponding to the retention time, and calculate the content of the component to be tested in the sample by the external standard method.

[0074] The chromatographic conditions for detecting ginsenosides were as follows: Agela Promosil C 18 Column (250mm×4.6mm, 5μm). Mobile phase A is acetonitrile solution, and mobile phase B is aqueous solution. The volume flow rate is set to 1mL / min, column temperature is 35℃, absorption wavelength is 203nm, injection volume is 20μL, mobile phase A is acetonitrile solution, mobile phase B is aqueous solution, gradient elution program is: 0-20min is 20% A; 20-45min is 20-46% A; 45-55min is 46-55% A; 55-60min is 55% A; 60-61min is 55-20A%; 61-75min is 20% A.

[0075] The chromatographic conditions for the detection of gingerol and shogaol are as follows: Agela Promosil C 18 Column (250mm×4.6mm, 5μm). Column temperature: 30℃; flow rate: 1.0mL / min. Wavelength: 280nm; injection volume: 10μL. Mobile phase A is acetonitrile solution, mobile phase B is aqueous solution, gradient elution program: 0min 55% A; 0-6min 65-80% A; 30-35min 80-90% A; 35-40min 90-55% A.

[0076] Content (%) = Ax*Cr / Ar*Cx;

[0077] Ax: sample peak area; Cr: reference substance concentration; Ar: reference substance peak area; Cx: sample concentration.

[0078] Wistar rats were purchased from Liaoning Changsheng Animal Co., Ltd. (Liaoning) and maintained in a SPF environment with a 12 h / 12 ​​h light / dark alternation.

[0079] Cisplatin for injection (lyophilized) was purchased from Qilu Pharmaceutical (Hainan) Co., Ltd.

[0080] IEC-6 cells were provided by Dalian Meilun Biotechnology Co., Ltd.

[0081] The 5-HT test kit was provided by Jiangsu ELISA Biotechnology Co., Ltd.

[0082] The cell viability detection kit was provided by Dalian Meilun Biotechnology Co., Ltd.

[0083] Reference substances: Ginsenosides Rg1, Re, Rc, Rh4, Rb1, Rg2, Rk3, Rh1, Rb2, F4, Rg6, Rd, S-Rg3, R-Rg3, Rk1 and Rg5, 6-gingerol, 8-gingerol, 10-gingerol and 6-shogaol were purchased from Chengdu Purifa Co., Ltd. with a purity of >98%.

[0084] Ginseng and ginger combination (GG), blank group (Control), model group (Model), ginseng group (GY), ginger group (JY), ginseng and ginger combination low-dose group (GGL) and ginseng and ginger combination high-dose group (GGH).

[0085] Ginseng and ginger were purchased from Yanji West Market.

[0086] Example 1

[0087] In order to better increase the content of ginsenosides and gingerols in GG, the parameters of the GG preparation method were optimized. The specific scheme is as follows:

[0088] 1. Effects of different ginseng-ginger ratios on the contents of ginsenosides and gingerols in GG.

[0089] 6g of ginseng and 180g of ginger were mixed, and distilled water 20 times the total weight of the two was added, and the mixture was soaked for 24 hours, and then heated and beaten at 100°C for 45 minutes, cooled, vacuum dried, and ground into powder to obtain GG.

[0090] GG was prepared by changing the ratio of ginseng to ginger, and the composition and content of prototype ginsenosides, rare ginsenosides and gingerol components in the obtained GG were detected. The results are shown in Tables 1 and 2.

[0091] In Table 1, the content of each ginsenoside is the percentage of each ginsenoside component in the total mass of ginseng.

[0092] In Table 2, the content of each gingerol is the percentage of each gingerol in the total weight of ginger.

[0093] Table 1 Effects of different ratios of ginseng and ginger on the composition and content of ginsenosides in GG (by mass percentage)

[0094]

[0095] Note: Bold fonts represent rare ginsenosides. The “rare ginsenosides” in the table are the sum of the rare ginsenosides represented by bold fonts. Prototype ginsenosides are the sum of the ginsenosides not represented by bold fonts in the table.

[0096] Table 2 Effects of different ratios of ginseng and ginger on the composition and content of gingerol in GG (by mass percentage)

[0097]

[0098]

[0099] 2. Effect of different beating time on the content of ginsenoside and gingerol in GG

[0100] 6g of ginseng and 180g of ginger were mixed, and distilled water 20 times the total mass of the two was added. After soaking for 24 hours, the mixture was heated to 100°C, and the mixture was beaten for 30, 45, 60, and 75 minutes, and cooled. After vacuum drying, the mixture was ground into powder to obtain GG. The types and contents of ginsenosides and gingerols in the obtained GG were detected. The types and contents of ginsenosides and gingerols in GG are shown in Tables 3 and 4.

[0101] In Table 3, the content of each ginsenoside is the percentage of each ginsenoside component in the total mass of ginseng.

[0102] In Table 4, the content of each gingerol is the percentage of each gingerol in the total weight of ginger.

[0103] Table 3 Effect of beating time on the composition and content of ginsenosides in GG (by mass percentage)

[0104]

[0105]

[0106] Note: Bold fonts represent rare ginsenosides. The “rare ginsenosides” in the table are the sum of the rare ginsenosides represented by bold fonts. Prototype ginsenosides are the sum of the ginsenosides not represented by bold fonts in the table.

[0107] Table 4 Effect of different beating time on the composition and content of gingerol in GG (by mass percentage)

[0108]

[0109] The results in Tables 1-4 show that when the ratio of ginseng to ginger is 1:10-1:60 and the heating time is 30-75 minutes, the content of rare ginsenosides is greatly increased, the content of Rh4 is above 20%, the content of total gingerol is 2.5-9%, the content of total ginsenosides is between 15-55%, and the content of rare ginsenosides accounts for more than 60% of the total ginsenosides.

[0110] When the ratio of ginseng to ginger is 1:30-1:60 and the beating time is 45-60min, the content of total ginsenosides is 30-55%, the content of rare ginsenosides accounts for more than 70% of the total ginsenosides, and the content of Rh4 is more than 50-70%; the content of total gingerol is 3.5-9%.

[0111] When the ratio of ginseng to ginger is 1:30 and the beating time is 45-60min, the content of total ginsenosides is 30-55%, the content of rare ginsenosides accounts for more than 75% of the total ginsenosides, and the content of Rh4 is 60-70%; the content of total gingerol is 5-9%.

[0112] Therefore, the preferred ratio of ginseng to ginger in the present invention is 1:30, and the beating time is 45 minutes. At this time, the content of total ginsenosides, the content of rare ginsenosides, and the content of Rh4 all reach the highest.

[0113] Example 2 Preparation of GG:

[0114] Ginseng puree: add 20 times the total mass of distilled water to 6g of ginseng, soak for 24 hours, heat to 100°C, beat for 45 minutes, cool, vacuum dry and grind to obtain ginseng group samples.

[0115] Ginger puree: Add 20 times the total weight of distilled water to 180g of fresh ginger, heat to 100°C, beat for 45min, cool, vacuum dry and grind to obtain ginger group samples.

[0116] Ginseng ginger composition (i.e. ginseng ginger puree): 6g ginseng and 180g fresh ginger were mixed, 20 times the total weight of distilled water was added, soaked for 24 hours, heated to 100°C, beaten for 45 minutes, cooled, vacuum dried and ground to obtain ginseng ginger composition (GG).

[0117] Example 3 Detection of ginsenoside types of GG in Example 2.

[0118] The LC-MS detection method was used to conduct a qualitative analysis of GG. The specific operation was as follows:

[0119] Measurement conditions: ACQUITY UPLC BEH C 18Chromatographic column (100×2.1mm, 1.7μm) was used for separation. Mobile phase A was 0.1% formic acid water, and mobile phase B was acetonitrile. The elution system was a linear gradient: 0-3min, 19-22% B; 3-6min, 22-30% B; 6-9min, 30-32% B; 9-16min, 32-35% B; 6-18min, 35-55% B; 18-22min, 55% B; 22-26min, 55-70% B; 26-29min, 70-90% B; 29-31min, 90-19% B; 31-33min, 19% B. Volume flow rate: 0.35mL / min; injection volume: 1μL; column temperature: 35℃; sample temperature: 19℃.

[0120] The collected raw mass spectrometry data were viewed and analyzed using Xcalibur 3.2 data software. Within the retention time range of 0-33min, the difference between the theoretical molecular weight and the actual measured value was ensured to be within 5ppm; the signal-to-noise ratio was set to 3; the retention time drift value was 0.3min, and within the full scan range of 80-1200m / z, combined with the MS / MS fragmentation of the first two ions, the structural identification of the auxiliary components, and the secondary spectrum of the molecular ion peak was screened. The detected mass spectrum fragments were then matched and identified with the database for compound matching.

[0121] LC-MS results showed that a total of 20 ginsenoside components were identified in the experiment, and the difference between the theoretical molecular weight and the actual measured value was within 5 ppm. The results showed that the composition was rich in rare saponins.

[0122] We speculate that acid hydrolysis of prototype saponins under the action of phenolic substances can promote the chemical transformation reaction of saponins and help to further improve the value of the composite product. The mass spectrometry results are shown in Table 5.

[0123] Table 5. LC-MS identification results of GG

[0124]

[0125]

[0126] Example 4 Determination of saponin content in GG of Example 2.

[0127] In order to verify the effective components of GG described in the present invention, the saponin content in GG was detected by HPLC test method.

[0128] Preparation of reference substances: Weigh 16 ginsenosides Rg1, Re, Rc, Rh4, Rb1, Rg2, Rk3, Rh1, Rb2, F4, Rg6, Rd, S-Rg3, R-Rg3, Rk1 and Rg5 reference substances respectively, dissolve them in chromatographic methanol and make up to volume to prepare standard solutions.

[0129] Sample preparation: After freeze-drying the samples of the ginseng group and the combination group in Example 2, 400 mg was accurately weighed, and an ethanol-water solution was added for ultrasonic-assisted extraction, centrifuged, and the supernatant was collected. The operation was repeated once. The combined supernatant was dried and dissolved in 10 mL of chromatographic grade methanol, filtered through a 0.45 μm organic microporous filter membrane and passed into a sample injection vial with a built-in tube for analysis and detection.

[0130] Chromatographic conditions:

[0131] HPLC analysis was performed on a L-2200 Hitachi HPLC system equipped with a Hitachi 1430DAD detector, 1310 column oven, 1210 injection tray, and 1110 pump. Agela Promosil C 18 Column (250mm×4.6mm, 5μm). Mobile phase A is acetonitrile solution, and mobile phase B is aqueous solution. The volume flow rate is set to 1mL / min, column temperature is 35℃, absorption wavelength is 203nm, injection volume is 20μL, mobile phase A is acetonitrile solution, mobile phase B is aqueous solution, and elution gradient process is: 0-20min is 20% A; 20-45min is 20-46% A; 45-55min is 46-55% A; 55-60min is 55% A; 60-61min is 55-20A%; 61-75min is 20% A.

[0132] Table 6. Average values ​​of PPD and PPT saponin contents (n=3)

[0133] PPD <![CDATA[Rb1]]> Rc <![CDATA[Rb2]]> Rd <![CDATA[S-Rg3]]> <![CDATA[R-Rg3]]> <![CDATA[Rk1]]> <![CDATA[Rg5]]> Ginseng 53.07±0.03 31.47±0.00 7.29±0.01 11.77±0.00 5.17±0.02 0.13±0.00 6.08±0.00 5.54±0.00 Composition 35.30±0.02 21.04±0.01 13.78±0.01 10.00±0.00 17.50±0.00 0.13±0.00 6.94±0.01 2.68±0.00 PPT <![CDATA[Rg1]]> Re <![CDATA[Rg2]]> <![CDATA[Rh1]]> <![CDATA[Rg6]]> F4 <![CDATA[Rk3]]> <![CDATA[Rh4]]> Ginseng 38.35±0.01 58.09±0.02 44.85±0.01 15.14±0.00 0.92±0.00 0.00±0.00 7.83±0.01 175.43±0.08 Composition 19.85±0.01 18.06±0.01 13.90±0.01 0.91±0.01 7.81±0.01 0.47±0.00 11.70±0.01 302.47±0.09

[0134] After the composition sample is prepared, it is tested under the above chromatographic conditions to obtain the peak area of ​​the reference substance, and then the peak area of ​​the sample corresponding to the retention time is measured, and the content of the component to be tested in the sample is calculated by the external standard method.

[0135] Saponin content (%) = Ax*Cr / Ar*Cx;

[0136] Ax: sample peak area; Cr: reference substance concentration; Ar: reference substance peak area; Cx: sample concentration.

[0137] Under the preparation conditions of the present invention, the C-20 position of the PPD-type prototype saponins Rc, Rb1 and Rb2 is deglycosylated under the action of acid to generate Rd, and Rd is further hydrolyzed to S-Rg3. Under these preparation conditions, S-Rg3 will not be further converted into Rg5 and Rk1, so this process can achieve efficient enrichment of the active ingredient S-Rg3. The C-20 position of the PPT-type prototype saponin Re is deglycosylated under the action of acid to generate Rg1 and Rg2. Under these preparation conditions, Rg1 is further dehydrated and rhamnose-converted into rare saponins F4 and Rh1, respectively, and deglycosylated through two pathways to generate rare saponin Rh4, achieving an ultra-high conversion rate of the rare saponin Rh4.

[0138] Example 5

[0139] In order to verify the effective components of GG described in the present invention, the gingerol content in GG was detected by HPLC test method.

[0140] Preparation of standard solution: Accurately weigh 6-gingerol, 8-gingerol, 10-gingerol and 6-shogaol standards, dissolve in chromatography grade methanol, make up to volume as reference stock solution, and store in a -20℃ refrigerator away from light.

[0141] Sample preparation: After freeze-drying the ginger group and the combination group samples in Example 2, 400 mg of each sample was accurately weighed, added to an ethanol-water solution, ultrasonic-assisted extraction, centrifuged, and the supernatant was collected, and the operation was repeated once. The combined supernatant was dried and dissolved in 10 mL of chromatographic grade methanol, filtered through a 0.45 μm organic microporous filter membrane into a sample injection vial with a built-in tube for analysis and detection.

[0142] Chromatographic conditions: HPLC analysis was performed on a L-2200 Hitachi HPLC system equipped with a Hitachi 1430DAD detector, 1310 column oven, 1210 injection tray, and 1110 pump. Agela Promosil C 18 Column (250mm×4.6mm,5μm). Column temperature: 30℃; flow rate 1.0m L / min. Wavelength: 280Nm; injection volume: 10μL. The theoretical plate numbers of 6-gingerol, 8-gingerol, 10-gingerol and 6-shogaol are all greater than 15000. Mobile phase A is acetonitrile solution, mobile phase B is aqueous solution, and the program gradient changes are as follows: 0min is 55% A; 0-6min is 65-80% A; 30-35min is 80-90% A; 35-40min is 90-55% A.

[0143] After the GG sample was prepared, it was tested according to the optimized chromatographic conditions to obtain the peak area of ​​the reference substance, and then the peak area of ​​the sample corresponding to the retention time was measured, and the content of the component to be tested in the sample was calculated according to the external standard method.

[0144] Saponin content (%) = Ax*Cr / Ar*Cx; Ax: sample peak area; Cr: reference substance concentration; Ar: reference substance peak area; Cx: sample concentration.

[0145] Table 7. Determination results of gingerol content in ginger and GG composition (n=3)

[0146]

[0147] Example 6 GG's effect in treating vomiting

[0148] In order to verify the vomiting treatment effect of GG of the present invention, a rat pica model was induced by cisplatin to detect the improvement of GG (Example 2) on rat body weight, normal feed intake and kaolin intake.

[0149] Wistar rats were purchased from Liaoning Changsheng Animal Co., Ltd. (Liaoning, China) and maintained in an SPF-level environment with a 12-h light / dark cycle. They were adaptively raised for 3 days, and the shape of the kaolin feed was similar to the normal feed consumed by rats. Before the experiment, the rats were fed with kaolin for three days until they no longer gnawed on kaolin, and then entered the formal experiment. The rats were randomly divided into 6 groups, 8 in each group, Control group + ip saline, Model group + ip cisplatin 6 mg / kg, GY group + ig 0.27 g / kg + ip cisplatin 6 mg / kg, JY group + ig 7.28 g / kg + ip cisplatin 6 mg / kg, GGL group + ig GY 0.135 + JY 3.64 g / kg + ip cisplatin 6 mg / kg, GGH group + ig GY 0.27 + JY 7.28 g / kg + ip cisplatin 6 mg / kg, and the kaolin intake of each group was observed at 24 h, 48 h, and 72 h after administration.

[0150] The body weight of rats in each group injected with cisplatin after modeling showed a downward trend, but the weight loss of rats within 24 hours was not obvious, and there was no significant difference between the groups injected with cisplatin (P>0.5). Compared with the normal control group, the body weight of rats in each group showed a significant decrease 72 hours after cisplatin injection (P<0.05), indicating that cisplatin can cause a significant weight loss trend in rats. The body weight of each group showed a slight upward trend 72 hours after treatment, but there was no significant difference (P>0.5). The intake of kaolin in rats increased significantly 24 hours, 48 ​​hours and 72 hours after cisplatin administration.

[0151] Compared with the Model, the consumption of kaolin by rats in each group was significantly reduced 48 hours after cisplatin injection (P<0.05). Overall, the rats in each treatment group showed a decrease in feed intake after modeling. This may be related to the delayed gastric emptying caused by the chemotherapy drug cisplatin in rats. Figure 1GG treatment can alleviate cisplatin-induced vomiting and anorexia.

[0152] In summary, GG can effectively relieve chemotherapy-induced nausea and vomiting, and its low price is of great significance in alleviating the economic burden of patients with chemotherapy-induced nausea and vomiting. It is worthy of further promotion and use in clinical practice, but its mechanism of action needs further research and exploration.

[0153] Example 7 GG relieves vomiting

[0154] In order to verify the vomiting-relieving effect of GG of the present invention, the enzyme-linked immunosorbent assay (ELISA) was used to detect the effect of GG (Example 2) on the 5-HT content in serum and medulla oblongata 72 hours after modeling. After sodium pentobarbital anesthesia, each group of rats was blooded from the heart, centrifuged, serum was collected, packaged, and frozen in a -80°C ultra-low temperature refrigerator for standby use. The ELISA method was used to determine the 5-HT content in the serum and medulla oblongata of the rats. The dosing regimen was the same as in Example 6.

[0155] 72h after modeling, the 5-HT content in the Model group increased significantly. Compared with the Model group, the GGL and GGH groups were significantly better than the single use group 72h after administration, and there was a significant difference (P<0.05). Figure 2 .

[0156] The results suggest that GG has an anti-chemoemetic effect, and its mechanism of action may be related to the 5-HT pathway in the brain.

[0157] Example 8 GG in the treatment of vomiting

[0158] In order to verify the effect of GG in treating vomiting of the present invention, a protein immunoblotting experiment was used to detect the effect of GG (Example 2) on the content of 5-HT receptor-related proteins (5-HT3A, MAOA, TPH2 and TPH1) in the ileum and medulla oblongata 72 hours after modeling. The samples were subjected to protein denaturation, electrophoretic separation, electrotransfer and other operations, and the protein membrane was incubated with the corresponding primary antibody at 4°C overnight, with β-actin as a control for total protein. In the presence of horseradish peroxidase-coupled secondary antibodies, the ECL reagent was reacted, and the signal was observed in real time in Tanon4600. The image was processed and analyzed using Image J. The dosing regimen was the same as in Example 6.

[0159] GGH can reduce the protein expression of 5-HT3A. As the administration time of cisplatin increased, the expression of TPH2 protein in the medulla oblongata increased significantly compared with the Control, indicating that cisplatin can promote the synthesis of 5-HT by upregulating TPH content. Compared with the Model group, GG can significantly reduce the content of TPH2. Compared with the Control group, the content of MAO-A in the medulla oblongata of the Model group was reduced and inhibited, while GG increased the content of MAO-A to accelerate the metabolism of additional 5-HT caused by cisplatin administration. Compared with the Model group, the relative content of 5-HT3A protein in the medulla oblongata and ileum of the Model group increased significantly with the increase in the number of cisplatin administration, while the GG group significantly reduced the expression of 5-HT3A protein. The results are shown in Figure 3 .

[0160] In summary, the above results suggest that GGH has the effect of relieving chemotherapy-induced vomiting and can be used to treat chemotherapy-induced vomiting.

[0161] Example 9 GG Protective Effect on Gastric Mucosa

[0162] In order to verify whether the vomiting treatment effect of GG described in the present invention is related to the relief of gastrointestinal pathological tissue damage caused by cisplatin, the recovery of pathological damage of the gastric antrum and ileum by the composition (Example 2) was observed by histopathological staining. The gastric antrum and ileum of rats were taken and pre-cooled with 4% paraformaldehyde for 24 hours. After dehydration and paraffin embedding, the tissue was cut into 4μm slices for HE staining to observe the histological changes of the gastric antrum and ileum. The dosing regimen was the same as that of Example 6.

[0163] The results are as follows Figure 4 As shown in the results, compared with the Control group, the tissues after cisplatin administration showed obvious pathological changes, including damage to the surface layer of the gastric antrum and damage to the ileal mucosal structure; and obvious inflammatory infiltration was observed. Compared with the Model group, GG reduced the pathological damage of the gastric antrum and ileum, including reducing inflammatory infiltration and mucosal damage. The above results suggest that GG can alleviate cisplatin-induced gastrointestinal damage by reducing pathological damage.

[0164] Example 10GG improves gastric emptying

[0165] In order to verify whether the vomiting-relieving effect of GG described in the present invention is related to the slow gastric emptying caused by cisplatin, a mouse gastric block model was established by intraperitoneal injection of cisplatin to verify the effect of GG (Example 2) on gastric emptying rate and small intestinal propulsion rate.

[0166] KM mice were purchased from the Experimental Animal Center of Yanbian University (Jilin, China), maintained in an SPF environment with a 12-h light / dark cycle, and were adaptively raised for 3 days. The mice were randomly divided into 6 groups (N=10): Control group (ip saline), Model group (ip cisplatin 3 mg / kg), GY group (ig 0.39 g / kg + ip cisplatin 3 mg / kg), JY group (ig 10.4 g / kg + ip cisplatin 3 mg / kg), GGL group (ig GY 0.195 + JY 5.2 g / kg + ip cisplatin 3 mg / kg), GGH group (ig GY 0.39 + JY 10.4 g / kg + ip cisplatin 3 mg / kg). After the experiment, the mice were anesthetized with sodium pentobarbital.

[0167] Compared with the Control group, the gastric remnant rate of the animals in the Model group increased significantly (P<0.01). Compared with the Model group, the GGH group could significantly reduce the gastric remnant rate (P<0.01). There was no statistically significant difference between the GGH group and the blank group (P>0.05), indicating that the improvement effect of the GGH group on the gastric emptying disorder caused by cisplatin can basically recover. Compared with the Control group, the small intestinal propulsion rate of the animals in the Model group increased significantly (P<0.001). Compared with the Model group, GGL had a significant inhibitory effect on the hyperpropulsion of the small intestine of the model animals (P<0.05), while the GGH group had a certain inhibitory trend on the hyperpropulsion of the small intestine. The results are shown in Figure 5 .

[0168] Example 11 Protective effects of the ginseng group and the combination group on IEC-6

[0169] To verify the effects of ginseng puree and composition group GG on rat intestinal epithelial cells (IEC-6) in Example 2 of the present invention, IEC-6 were cultured in Dulbecco's modified medium containing 10% fetal bovine serum and 1% penicillin-streptomycin. A humidified incubator containing 5% CO2 at 37°C was provided for IEC-6 cells.

[0170] Cell counting kit-8 (CCK-8) was used for drug concentration screening and cell viability determination. IEC-6 cells were plated at 2×10 3 The cells were seeded in a 96-well plate at a density of 10 cells. After overnight incubation, the medium was replaced with ginseng puree (20 μM) and the composition (20 μM). One hour later, 20 μM cisplatin was administered to induce chemotherapeutic cell damage. During the 24-hour incubation in the dark, 10% CCK-8 reagent diluted with DMEM was added to each well. After 4 hours of incubation, the absorbance of each well was measured at 450 nm.

[0171] The results showed that both the ginseng group and the combination group could restore the cell viability of IEC-6 cells after cisplatin treatment to varying degrees, among which the combination group had the strongest protective effect. Figure 6 .

[0172] Example 12 Protective effects of ginger group and combination group on IEC-6

[0173] To verify the effects of the ginger group and the composition group on IEC-6 cells in Example 2 of the present invention, IEC-6 cells were cultured in Dulbecco's modified medium containing 10% fetal bovine serum and 1% penicillin-streptomycin. A humidified incubator containing 5% CO2 at 37°C was provided for IEC-6 cells.

[0174] Cell counting kit-8 (CCK-8) was used for drug concentration screening and cell viability determination. IEC-6 cells were plated at 2×10 3 The cells were seeded in a 96-well plate at a density of 10 cells. After overnight incubation, the medium was replaced with ginger puree and the composition at a concentration of 20 μM. After 1 h, 20 μM cisplatin was given to induce chemotherapeutic cell damage. During the 24 h incubation in the dark, 10% CCK-8 reagent diluted in DMEM was added to each well. After 4 h of incubation, the absorbance of each well was measured at 450 nm.

[0175] The results showed that compared with the ginger group, the combination group could significantly restore the cell viability of IEC-6 cells after cisplatin treatment. Figure 7 .

[0176] Example 13 Protective effects of different gingerols on IEC-6

[0177] To verify the effects of gingerol and shogaol in the GG of the present invention on IEC-6 cells, IEC-6 cells were cultured in Dulbecco's modified medium containing 10% fetal bovine serum and 1% penicillin-streptomycin. A humidified incubator containing 5% CO2 at 37°C was provided for IEC-6 cells.

[0178] Cell counting kit-8 (CCK-8) was used for drug concentration screening and cell viability determination. IEC-6 cells were plated at 2×10 3 The cells were seeded in a 96-well plate at a density of 10 cells. After overnight incubation, the medium was replaced with 2.5 μM 6-gingerol, 6-shogaol, 8-gingerol, and 10-gingerol. One hour later, 20 μM cisplatin was given to induce chemotherapeutic cell damage. During the 24-hour incubation in the dark, 10% CCK-8 reagent diluted in DMEM was added to each well. After 4 hours of incubation, the absorbance of each well was measured at 450 nm.

[0179] The results showed that gingerols in ginger could restore the cell viability of IEC-6 cells after cisplatin treatment. Figure 8 .

[0180] Example 14 GG's efficacy against motion sickness

[0181] Verification of the GG of the present invention The behavioral changes of rats in each group were detected by heterophilic kaolin experiment, in order to verify the pharmacodynamic effect of GG (Example 2) on motion sickness.

[0182] SD rats were purchased from the Experimental Animal Center of Yanbian University (Jilin, China), maintained in an SPF-level environment with a 12-h light / dark cycle, and were adaptively fed for 3 days. The rats were randomly divided into 6 groups (N=6): Control group + (ip saline), Model group + (ip cisplatin 6 mg / kg), GY group + (ig0.27 g / kg + ip cisplatin 6 mg / kg), JY group + (ig7.28 g / kg + ip cisplatin 6 mg / kg), GGL group + (igGY 0.135 + JY 3.64 g / kg + ip cisplatin 6 mg / kg), GGH group + (igGY 0.27 + JY 7.28 g / kg + ip cisplatin 6 mg / kg). The rats in each group were housed in a single cage. After 3 days of adaptive feeding, a certain amount of feed and kaolin (separately placed) were given every day from the 4th day, and the 24-h kaolin intake of the rats was recorded for 3 days before and after modeling. The experimental results were evaluated by the difference in the total amount of kaolin intake of each rat in the 3 days before and after modeling, that is, the total amount of kaolin intake of each rat in the 3 days after modeling was subtracted from the total amount of kaolin intake in the 3 days before modeling, and statistical analysis was performed. Compared with the Control group, the rats in the Model group began to eat kaolin after modeling, the kaolin intake increased significantly, and the difference in the total amount of kaolin intake before and after modeling increased significantly; compared with the Model group, the kaolin craving behavior of rats in the low- and high-dose GG groups was significantly reduced, and the difference in the total amount of kaolin intake in 3 days was significantly reduced. The results are as follows Fig. 9 As shown, it is suggested that GG has a certain anti-motion sickness effect.

[0183] The above embodiments are merely examples for the purpose of clear explanation, and are not intended to limit the implementation methods. For those skilled in the art, other different forms of changes or modifications can be made based on the above description. It is not necessary and impossible to list all the implementation methods here. The obvious changes or modifications derived therefrom are still within the scope of protection of the present invention.

Claims

1. A composition for relieving nausea and vomiting, characterized in that: The composition is prepared by using ginseng and ginger as raw materials, and mixing and beating after adding water; the mass ratio of the ginseng to ginger is 1:10-1:60, preferably 1:30-1:60; the mass ratio of the mixed raw material of ginseng, ginger and water is 1:10-1:

40.

2. The composition according to claim 1, characterized in that The composition contains total ginsenosides and total gingerols, wherein the total ginsenoside content is 15-55%, the total gingerol content is 2.5-9%, and the rare ginsenoside content accounts for more than 60% of the total ginsenoside content; preferably, the total ginsenoside content is 30-55%, the total gingerol content is 3.5-9%, and the rare ginsenoside content accounts for more than 70% of the total ginsenoside content; preferably, the total ginsenoside content is 30-55%, the total gingerol content is 5-9%, and the rare ginsenoside content accounts for more than 75% of the total ginsenoside content.

3. The composition according to claim 1 or 2, characterized in that The total ginsenosides contain the following components: ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, ginsenoside Rc, ginsenoside Rg2, ginsenoside Rb2, ginsenoside Rh1, ginsenoside Rd, ginsenoside Rg6, ginsenoside F4, ginsenoside Rk3, ginsenoside Rh4, ginsenoside S-Rg3, ginsenoside R-Rg3, ginsenoside Rk1, ginsenoside Rg5; the weight percentage composition of each component is as follows: ginsenoside Rg1 0.51-1.99%, ginsenoside Re 0.78-3.63%, ginsenoside Rb1 2.16-3.56%, ginsenoside Rc0.61-2.70%, ginsenoside Rg2 0.73-1.87%, ginsenoside Rb2 0.34-1.68%, ginsenoside Rh1 0.01-0.29%, ginsenoside Rd 0.21-1.63%, ginsenoside Rg6 0.01-2.11%, ginsenoside F4 0.05-2.27%, ginsenoside Rk3 0.01-1.19%, ginsenoside Rh4 5.54-33.25%, ginsenoside S-Rg3 0.07-1.75%, ginsenoside R-Rg3 0.01-0.19%, ginsenoside Rk1 0.08-0.69%, ginsenoside Rg5 0.02-2.12%; Preferably, The weight percentage composition of each component is as follows: ginsenoside Rg1 0.51-1.99%, ginsenoside Re 0.78-2.72%, ginsenoside Rb1 2.24-3.56%, ginsenoside Rc 0.61-2.70%, ginsenoside Rg2 0.73-1.43%, ginsenoside Rb2 0.34-1.68%, ginsenoside Rh1 0.01-0.29%, ginsenoside Rd 0.21-1.63%, ginsenoside Rg6 0.01-1.13%, ginsenoside F4 0.05-1.12%, ginsenoside Rk3 0.38-1.19%, ginsenoside Rh4 19.53-33.25%, ginsenoside S-Rg3 0.11-1.75%, ginsenoside R-Rg3 0.01-0.04%, ginsenoside Rk1 0.23-0.69%, ginsenoside Rg5 0.17-2.12%; The total gingerol contains the following components: 6-shogaol, 6-gingerol, 8-gingerol, and 10-gingerol; The weight percentage of each component is as follows: 6-shogaol 0.98-3.66%, 6-gingerol 0.98-2.17%, 8-gingerol 0.06-1.53%, 10-gingerol 0.16-1.54%; Preferably, The weight percentage of each component is as follows: 1.23-3.66% of 6-shogaol, 1.25-2.17% of 6-gingerol, 0.72-1.32% of 8-gingerol, and 0.34-1.54% of 10-gingerol.

4. A method for preparing a composition according to any one of claims 1 to 3, characterized in that: The ginseng is mixed with fresh ginger, soaked in water, and mixed and beaten at 60-100° C. for 30-75 min to obtain the product; preferably, the ginseng is mixed with fresh ginger, soaked in water, and mixed and beaten at 90-100° C. for 45-60 min.

5. A pharmaceutical preparation comprising the composition of any one of claims 1 to 5 and a pharmaceutically acceptable carrier or excipient.

6. Use of the composition according to any one of claims 1 to 3 or the preparation according to claim 5 in the preparation of a medicament for relieving / treating nausea and vomiting.

7. The use according to claim 6, characterized in that: The nausea and vomiting include nausea and vomiting caused by chemotherapy drugs, nausea and vomiting caused by radiotherapy, postoperative vomiting, vomiting during pregnancy, motion sickness, or vomiting caused by gastrointestinal damage, inflammatory diseases, and anorexia.

8. The use according to claim 7, characterized in that: The chemotherapy drug is one or more of cisplatin, paclitaxel, etoposide, teniposide, daunorubicin, doxorubicin, idarubicin, cyclophosphamide, mitoxantrone, aclarubicin, homoharringtonine, methotrexate, vincristine, and vindesine.

9. Use of the composition according to any one of claims 1 to 3 or the preparation according to claim 5 in the preparation of medicines and foods for assisting in protecting gastric mucosa.

10. The composition according to any one of claims 1 to 3, characterized in that Ginger increased the content of rare saponins in ginseng, and ginseng increased the content of total gingerols in ginger.

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