AU2021246790A1

AU2021246790A1 - Anti-tumor composition containing rare ginsenosides Rk2, CK, and PPT

Info

Publication number: AU2021246790A1
Application number: AU2021246790A
Authority: AU
Inventors: Yangfang DONG; Zhiguang DUAN; Daidi Fan; Jianya YAN
Original assignee: Shaanxi Giant Biotechnology Co Ltd
Current assignee: Shaanxi Giant Biotechnology Co Ltd
Priority date: 2020-03-31
Filing date: 2021-03-31
Publication date: 2022-12-08
Anticipated expiration: 2041-03-31
Also published as: CN111265536B; WO2021197372A1; CN111265536A; AU2021246790B2

Abstract

An anti-tumor composition containing the rare ginsenosides Rk2, CK, and PPT, taking a therapeutically effective dose of ginsenoside Rk2, ginsenoside CK, and ginsenoside PPT as active ingredients, and further containing a pharmaceutically acceptable carrier. In the rare ginsenoside pharmaceutical composition, the mass ratio between the ginsenoside Rk2, the ginsenoside CK, and the ginsenoside PPT is: 1:(1-2):(1-2). Research has shown that when the mass ratio between the ginsenoside Rk2, the ginsenoside CK, and the ginsenoside PPT is a particular ratio, in the present anti-tumor composition, not only does a synergistic effect occur between the ginsenoside Rk2, the ginsenoside CK, and the ginsenoside PPT, but a synergistic effect also occurs between any two of the three.

Description

ANTI-TUMOR COMPOSITION CONTAINING RARE GINSENOSIDES RK2,

CK, AND PPT

TECHNICAL FIELD The application relates to the field of biomedicine, in particular to a rare ginsenoside

Rk2/CK/PPT composition with anti-tumor activity and synergistic effect.

BACKGROUND OF THE INVENTION

Cancer, as one of the most serious and malignant diseases in the world, has seriously threatened

human health and quality of life. Its incidence and mortality are second only to cardiovascular and cerebrovascular diseases, and it is one of the most important causes of human death. The national

cancer incidence situation is grim, and the incidence and mortality of different cancers such as lung

cancer, breast cancer, and colon cancer continue to rise. Each year, there are about 4 million new cancer cases and about 2.5 million deaths due to cancer. At present, the relatively popular treatment

methods for cancer include chemotherapy, radiation therapy and surgery. Although these traditional

treatment methods can alleviate the disease, they have serious side effects and bring great physical and mental pain to patients. Therefore, the anti-tumor properties of natural medicines have gained

increasing attention.

Natural medicines are an important resource with great potential, and can be used to treat diseases such as tumor. Panax plants, such as ginseng, notoginseng, and American ginseng, as a class

of precious Chinese medicinal materials, have been used to treat various diseases, such as cancer. At

present, there are more than 60 kinds of ginsenosides founded, most of which belong to the prototype ginsenosides, including R, Rbl, Rb2, Rb2, Rd, Re, Rf, Rgl, etc. They are easily soluble in water

and are the main components of panax plants exerting various pharmacological effects. They have

pharmacological effects such as soothing the nerves, anti-oxidation, and regulating immune function, but they have not shown a good anti-tumor effect.

For rare ginsenoside monomers (mainly including Rg3, Rh2, Rkl, Rg5, Rk3, Rh4, CK, Rk2,

Rh3, PPD, PPT, etc), their content in ginseng plants is very small, and they can only exist after processing and are not easy to obtain. They have poor water solubility, and only show good solubility

in organic solvents such as ethanol and ethyl acetate. The above-mentioned rare ginsenoside monomers usually exhibit different anti-tumor effects due to the influence of the sugar in the structure, and the order of the strength of their anti-tumor activities is: aglycone > monoglycoside > diglycoside > triglycoside > tetraglycoside. At present, a large number of literatures show that rare ginsenoside monomers have obvious anti-tumor effects, which can block cycle distribution of tumor cells, induce tumor cell apoptosis, inhibit tumor angiogenesis, etc. However, the existing single rare ginsenoside monomer, such as any of the above-mentioned more than a dozen rare ginsenoside monomers, does not show a particularly significant anti-tumor effect.

SUMMARY OF THE INVENTION The purpose of the present application is to provide a rare ginsenoside composition based on rare ginsenoside Rk2, ginsenoside CK and ginsenoside PPT, which is synergistic and has anti-tumor effect. In addition, another purpose of the present application is to provide the use of the above-mentioned rare ginsenoside composition in the preparation of an anti-tumor medicament. In the present application, among more than a dozen rare ginsenoside monomers, such as Rg3, Rh2, Rkl, Rg5, Rk3, Rh4, CK, Rk2, Rh3, PPD, PPT, etc., were applied to tumor cells after combined in twos or threes, and related detections were performed. Among them, MTT method was used to detect their inhibitory effect on tumor cells and the combination index was calculated; flow cytometry was used to detect their effect on the distribution of tumor cell cycle; AV/PI kit was used to detect their apoptotic effect on tumor cells and the apoptosis rate was determined; and Western blotting was used to detect their effect on the expression of apoptosis-related proteins in tumor cells. The detection results of flow cytometry showed that the rare ginsenoside Rk2/CK/PPT composition blocked significantly more tumor cells in the GI phase than that of the group applied with a single ingredient, showing a strong synergistic effect. The detection results with AV/PI kit showed that the rare ginsenoside Rk2/CK/PPT composition induced tumor cell apoptosis, and its early apoptosis rate was significantly greater than that of the group applied with a single ingredient. The detection results of Western blotting showed that ginsenoside Rk2/CK/PPT composition promoted the activation of Caspase-3 and PARP proteins, down-regulated the expression of Bcl-2 protein, and finally induced tumor cell apoptosis. A large number of experimental results showed that the combination of ginsenoside Rk2, ginsenoside CK and ginsenoside PPT in a specific ratio not only had synergistic anti-tumor effect, but also exhibited low toxicity. Thus the present application was achieved. That is, the present application includes:

1. A rare ginsenoside pharmaceutical composition with anti-tumor effect (rare ginsenoside

pharmaceutical composition 1 of the present application), comprising therapeutically effective

amounts of ginsenoside Rk2, ginsenoside CK and ginsenoside PPT as active ingredients, and a pharmaceutically acceptable carrier,

wherein in the rare ginsenoside pharmaceutical composition, the weight ratio of ginsenoside

Rk2, ginsenoside CK and ginsenoside PPT is: 1: (1 to 2): (1 to 2). 2. The rare ginsenoside pharmaceutical composition according to item 1, wherein in the rare

ginsenoside pharmaceutical composition, the weight ratio of ginsenoside Rk2, ginsenoside CK and

ginsenoside PPT is: 1: (1 to 1.1): (1 to 1.1). 3. The rare ginsenoside pharmaceutical composition according to item 1 or 2, wherein in the

rare ginsenoside pharmaceutical composition, the weight ratio of ginsenoside Rk2, ginsenoside CK

and ginsenoside PPT is 1: 1: 1. 4. The rare ginsenoside pharmaceutical composition according to any one of items 1-3, wherein

the rare ginsenoside pharmaceutical composition does not comprise other ginsenosides.

5. The rare ginsenoside pharmaceutical composition according to any one of items 1-4, wherein the rare ginsenoside pharmaceutical composition comprises ginsenoside Rk2, ginsenoside CK and

ginsenoside PPT as the only anti-tumor active ingredients.

6. Use of a composition comprising ginsenoside Rk2, ginsenoside CK and ginsenoside PPT in the preparation of an anti-tumor medicament, wherein in the composition, the weight ratio of

ginsenoside Rk2, ginsenoside CK and ginsenoside PPT is: 1: (1 to 2): (1 to 2);

and the anti-tumor medicament comprises ginsenoside Rk2, ginsenoside CK and ginsenoside PPT as active ingredients.

7. The use according to item 6, wherein in the composition, the weight ratio of ginsenoside Rk2,

ginsenoside CK and ginsenoside PPT is: 1: (1 to 1.1): (1 to 1.1). 8. The use according to item 6 or 7, wherein in the composition, the weight ratio of ginsenoside

Rk2, ginsenoside CK and ginsenoside PPT is 1:1:1.

9. The use according to any one of items 6-8, wherein the composition does not comprise other ginsenosides.

10. The use according to any one of items 6-9, wherein the composition comprises ginsenoside

Rk2, ginsenoside CK and ginsenoside PPT as the only anti-tumor active ingredients. The inventor also found that under a specific ratio (for example, the weight ratio of ginsenoside

Rk2, ginsenoside CK and ginsenoside PPT is 1: (1 to 2): (1 to 2), preferably 1: (1 to 1.1): (1 to 1.1),

more preferably 1: 1: 1), in the rare ginsenoside pharmaceutical composition 1 of the present application, not only ginsenoside Rk2, ginsenoside CK and ginsenoside PPT are synergistic, but any two of the above three are also synergistic.

Therefore, the present application also includes:

11. A rare ginsenoside pharmaceutical composition with anti-tumor effect (rare ginsenoside pharmaceutical compositions 2-4 of the present application), comprising therapeutically effective

amounts of ginsenoside Rk2 and ginsenoside CK (or ginsenoside Rk2 and ginsenoside PPT, or

ginsenoside CK and ginsenoside PPT) as active ingredients, and a pharmaceutically acceptable carrier,

Rk2 to ginsenoside CK is 1: (1 to 2) (or the weight ratio of ginsenoside Rk2 to ginsenoside PPT is 1: (I to 2), or the weight ratio of ginsenoside CK to ginsenoside PPT is (I to 2): (1 to 2)).

12. The rare ginsenoside pharmaceutical composition according to item 11, wherein in the rare

ginsenoside pharmaceutical composition, the weight ratio of ginsenoside Rk2 to ginsenoside CK is 1: (I to 1.1) (or the weight ratio of ginsenoside Rk2 to ginsenoside PPT is 1: (1 to 1.1), or the weight

ratio of ginsenoside CK to ginsenoside PPT is ( to 1.1): (1 to 1.1)).

13. The rare ginsenoside pharmaceutical composition according to item I Ior 12, wherein in the rare ginsenoside pharmaceutical composition, the weight ratio of ginsenoside Rk2 to ginsenoside CK

is 1:1 (or the weight ratio of ginsenoside Rk2 to ginsenoside PPT is 1:1, or the weight ratio of

ginsenoside CK to ginsenoside PPT is 1:1). 14. The rare ginsenoside pharmaceutical composition according to any one of items I Ito 13,

wherein the rare ginsenoside pharmaceutical composition does not comprise other ginsenosides.

15. The rare ginsenoside pharmaceutical composition according to any one of items I Ito 14, wherein the rare ginsenoside pharmaceutical composition comprises ginsenoside Rk2 and

ginsenoside CK (or ginsenoside Rk2 and ginsenoside PPT, or ginsenoside CK and ginsenoside PPT)

as the only anti-tumor active ingredients. 16. Use of a composition comprising ginsenoside Rk2 and ginsenoside CK (or ginsenoside Rk2

and ginsenoside PPT, or ginsenoside CK and ginsenoside PPT) in the preparation of an anti-tumor

medicament, wherein in the composition, the weight ratio of ginsenoside Rk2 to ginsenoside CK is 1:(1 to 2) (or the weight ratio of ginsenoside Rk2 to ginsenoside PPT is 1: (1 to 2), or the weight ratio of ginsenoside CK to ginsenoside PPT is (I to 2): (1 to 2)); and the anti-tumor medicament comprises ginsenoside Rk2 and ginsenoside CK (or ginsenoside

Rk2 and ginsenoside PPT, or ginsenoside CK and ginsenoside PPT) as active ingredients. 17. The use according to item 16, wherein in the composition, the weight ratio of ginsenoside

Rk2 to ginsenoside CK is 1: (1 to 1.1) (or the weight ratio of ginsenoside Rk2 to ginsenoside PPT is

1: (1 to 1.1), or the weight ratio of ginsenoside CK to ginsenoside PPT is (I to 1.1): (1 to 1.1)). 18. The use according to item 16 or 17, wherein in the composition, the weight ratio of

ginsenoside Rk2 to ginsenoside CK (or ginsenoside Rk2 to ginsenoside PPT, or ginsenoside CK to

ginsenoside PPT) is 1: 1. 19. The use according to any one of items 16-18, wherein the composition does not comprise

other ginsenosides.

20. The use according to any one of items 16 to 19, wherein the composition comprises ginsenoside Rk2 and ginsenoside CK (or ginsenoside Rk2 and ginsenoside PPT, or ginsenoside CK

and ginsenoside PPT) as the only anti-tumor active ingredients.

In this specification, the rare ginsenoside pharmaceutical composition 1 of the present application and the rare ginsenoside pharmaceutical compositions 2-4 of the present application are

collectively referred to as the rare ginsenoside pharmaceutical composition of the present application

in some cases. The rare ginsenoside pharmaceutical composition of the present application was administered to

subcutaneous tumor-bearing mice, and its effects on tumor volume and body weight of the

tumor-bearing mice were detected and observed. The results showed that the group applied with a single ingredient could inhibit the increase of the tumor volume, and the rare ginsenoside

pharmaceutical composition of the present application showed obvious synergistic effect in

inhibiting tumor growth; there was no obvious difference in the body weight of nude mice in all groups, showing low toxicity characteristics. Therefore, the present application also relates to the use

of the rare ginsenoside pharmaceutical composition of the present application in the preparation of an

anti-tumor medicament. The tumor may be, for example, gastric cancer, liver cancer, or pancreatic cancer.

The rare ginsenoside pharmaceutical composition of the present application can be, for example,

an oral preparation or an injection. The oral preparation can be, for example, hard capsules, soft capsules, sustained or controlled release capsules, sugar-coated tablets, powders, granules, dropping pills, water-honeyed pills, syrups or oral liquids; the injection is solution form, suspension form, emulsion form or lyophilized powder. The rare ginsenoside pharmaceutical composition of the present application may comprise excipients or other pharmaceutically acceptable carriers. The excipient can be, for example, one or more of sodium hyaluronate, sodium alginate, chitosan or collagen.

BRIEF DESCRIPTION OF THE DRAWINGS

Figure 1 is a graph showing the effect of rare ginsenoside Rk2/CK/PPT composition 1,

composition 6, composition 9 and composition 12 on the distribution of colon cancer cell cycle. Figure 2 is a graph showing the apoptosis-inducing effect of rare ginsenoside Rk2/CK/PPT

composition 1, composition 6, composition 9 and composition 12 on colon cancer cells.

Figure 3 is a graph showing the effects of rare ginsenoside Rk2/CK/PPT composition 1 on the expression of Bel-2 family proteins, Caspase-3 and PARP proteins in colon cancer cells.

Figure 4 is a graph showing the inhibitory effect of rare ginsenoside Rk2/CK/PPT composition

1, composition 6, composition 9 and composition 12 on the colon cancer xenograft model. Figure 5 is a graph showing the effect of rare ginsenoside Rk2/CK/PPT composition 1 on

changes in water intake, food intake and body weight of healthy mice.

Figure 6 is a graph showing the effect of rare ginsenoside Rk2/CK/PPT composition 1 on liver function and kidney function of healthy mice.

In the above Figures 1-6, unless otherwise specified, no mark indicates no significant difference,

* indicates significant difference, P<0.05; ** indicates significant difference, P<0.01; and*** indicates significant difference, P<0.01 <0.001.

DETAIL DESCRIPTION OF THE INVENTION The present application will be described in detail below with reference to specific

embodiments. Unless conflicts exist, scientific terms in this specification have the meanings

commonly understood by those skilled in the art, and in case of conflict, the definitions in this specification shall prevail.

First, in one aspect, the present application provides a rare ginsenoside pharmaceutical

composition with anti-tumor effect (rare ginsenoside pharmaceutical composition of the present application, hereinafter also referred to as a pharmaceutical composition of the present application or a pharmaceutical composition). It comprises therapeutically effective amounts of ginsenoside Rk2, ginsenoside CK and ginsenoside PPT (or ginsenoside Rk2 and ginsenoside CK, or ginsenoside Rk2 and ginsenoside PPT, or ginsenoside CK and ginsenoside PPT) as active ingredients, and a pharmaceutically acceptable carrier,

Wherein, in this rare ginsenoside pharmaceutical composition, the weight ratio of ginsenoside

In the present specification, ginsenoside PPT (protopanaxatriol) refers to a compound

represented by the following chemical formula 1.

[Chemical formula 1]

OH HO

20

1

HO OH

The above-mentioned protopanaxatriol (PPT) is a known compound and can be prepared by a

method known in the art. For example, protopanaxatriol (PPT) can be prepared by enzymatic hydrolysis of ginsenoside Re.

In the present specification, ginsenoside C-K refers to a compound represented by the following

chemical formula 2.

[Chemical formula 2] OH 0

OH OOH O HO HO

The above-mentioned ginsenoside C-K is a known compound, and can be prepared by a method known in the art. For example, ginsenosides C-K can be prepared by pectinase-catalyzed hydrolysis of ginsenoside Rb1. In the present specification, ginsenoside Rk2 refers to a compound represented by the following chemical formula 3.

[Chemical formula 3]

HH OH

H HO H 5H

The above-mentioned ginsenoside Rk2 is a known compound, and can be prepared by a method known in the art. For example, ginsenoside Rk2 can be prepared by catalyzing ginsenoside Rbl at high temperature with an aqueous sodium hydroxide solution. The inventors found that the combination of ginsenoside Rk2, ginsenoside CK and ginsenoside PPT according to the ratio range defined in the present application not only show significant synergy in anti-tumor (CI value is less than 1, preferably CI value is less than 0.7), but also show low toxicity characteristics. The inventors also found that within the above ratio range, ginsenoside Rk2 and ginsenoside CK, or ginsenoside Rk2 and ginsenoside PPT, or ginsenoside CK and ginsenoside PPT are also synergistic and low in toxicity. Moreover, the inventors also found that if the contents of ginsenoside Rk2, ginsenoside CK and ginsenoside PPT in the pharmaceutical composition are not within the ratio range defined in the present application, there is no synergistic effect. Considering from the viewpoint of significant synergistic effect (CI value is less than 0.7), in the composition, the weight ratio of ginsenoside Rk2, ginsenoside CK and ginsenoside PPT is preferably 1: (1 to 1.1): (1 to 1.1), more preferably 1: 1: 1. In the pharmaceutical composition of the present application, other ginsenosides are either included or not. When other ginsenosides are included in the pharmaceutical composition of the present application, considering from the viewpoint of better synergistic effect, the contents of ginsenoside Rk2, ginsenoside CK and ginsenoside PPT (or ginsenoside Rk2 and ginsenoside CK, or ginsenoside Rk2 and ginsenoside PPT, or ginsenoside CK and ginsenoside PPT) are preferably 50 parts by weight or more, more preferably 60 parts by weight or more, more preferably 70 parts by weight or more, more preferably 80 parts by weight or more, more preferably 90 parts by weight or more, more preferably 95 parts by weight or more, more preferably 99 parts by weight or more, more preferably 100 parts by weight (that is, the pharmaceutical composition only comprises these three ginsenosides or only comprises the above-mentioned two ginsenosides), assuming that all the ginsenosides contained in the pharmaceutical composition are 100 parts by weight. The purity of ginsenoside Rk2 used in the pharmaceutical composition of the present application can be greater than 98%, the purity of ginsenoside CK can be greater than 98%, and the purity of ginsenoside PPT can be greater than 95%.

The content of all the ginsenosides can be determined by the vanillin method, and the contents of the ginsenosides Rk2, CK and PPT can be determined by the HPLC method.

The pharmaceutical composition of the present application can either comprise other anti-tumor

active ingredients or not comprise other anti-tumor active ingredients (that is, ginsenoside Rk2, ginsenoside CK and ginsenoside PPT (or ginsenoside Rk2 and ginsenoside CK, or ginsenoside Rk2

and ginsenoside PPT, or ginsenoside CK and ginsenoside PPT) are used as the only anti-tumor active

ingredients. On the other hand, the inventors found that the pharmaceutical composition of the present

application has a significant anti-tumor effect. Therefore, the present application also provides the

use of the pharmaceutical composition of the present application in the preparation of an anti-tumor medicament.

The pharmaceutical composition of the present application may comprise a pharmaceutically

acceptable carrier, such as an excipient. There are no special restrictions on the excipient in the pharmaceutical composition of the present application, for example, the excipients commonly used

in medicines or health products in the art can be used. Specifically, the excipient is starch, dextrin,

lactose, mannitol, sodium hypromellose, xanthan gum, protein sugar and the like. There are no special restrictions on the dosage form of the pharmaceutical composition of the

present application, for example, it may be an oral dosage form or an injection dosage form. The oral

dosage form can be a liquid dosage form or a solid dosage form. The oral dosage forms can be, for example, hard capsules, soft capsules, sustained or controlled release capsules, compressed tablets, sugar-coated tablets, powders, granules, dropping pills, water-honeyed pills, syrups or oral liquids; the injection dosage form can be, for example, solution form, suspension form, emulsion form or lyophilized powder. The administration method of the pharmaceutical composition for anti-tumor medicament can be, for example, oral administration, drip or injection.

When preparing a solid preparation for oral use, after adding an vehicle and optionally a binder,

adisintegrating agent, a lubricating agent, a coloring agent, a flavoring agent and the like to the main drug, it can be prepared into tablets, coated tablets, granules, fine granules, powders, capsules, and

the like according to a conventional method.

As an vehicle, for example, lactose, corn starch, white sugar, glucose, sorbitol, crystalline cellulose, silicon dioxide and the like can be used; as a binder, for example, polyvinyl alcohol, ethyl

cellulose, methyl cellulose, gum arabic, hydroxypropyl cellulose, hydroxypropyl methyl cellulose

and the like can be used; as a lubricating agent, for example, magnesium stearate, talc, silicon dioxide, and the like can be used; as a coloring agent, those allowed to be added to pharmaceuticals

can be used; as a flavoring agent, cocoa powder, menthol, aromatic acid, peppermint oil, borneol,

and cinnamon powder can be used. Of course, the above-mentioned tablets and granules can also be coated with a sugar coating, a gelatin coating, and other necessary outer coatings.

When preparing injections, pH regulators, buffers, suspending agents, solubilizers, stabilizers,

isotonic agents, preservatives and the like can be added to the main drug as required, and then intravenous, subcutaneous, and intramuscular injections can be prepared according to conventional

methods. At this time, if necessary, a lyophilized product can be prepared with a conventional

method. Examples of the suspending agent include methyl cellulose, Tween 80, hydroxyethyl cellulose,

acacia, tragacanth gum powder, sodium carboxymethyl cellulose, polyoxyethylene sorbitan

monolaurate, and the like. Examples of the solubilizer include polyoxyethylene hydrogenated castor oil, Tween 80,

niacinamide, polyoxyethylene sorbitan monolaurate, polyethylene glycol, castor oil fatty acid ethyl

ester, and the like. Moreover, examples of the stabilizer include sodium sulfite, sodium metasulfite, and the like;

examples of the preservative include methylparaben, ethylparaben, sorbic acid, phenol, cresol,

chlorocresol, and the like. The pharmaceutical composition of the present application is administered to a subject, where a tumor can be treated. The subject may be a mammal, such as a human, a rat, a rabbit, a sheep, a pig, a cow, a cat, a dog, a monkey, etc., and preferably a human.

The pharmaceutical composition of the present application can be administered orally or non-orally. The administration amount varies depending on the degree of symptoms, patient age, sex,

body weight, difference in sensitivity, administration method, administration period, administration

interval, the nature of the pharmaceutical preparation, the type of active ingredient, etc. There is no special restrictions, but for adults (body weight 60 Kg), it is usually 1 pg-30000 mg, preferably 10

pg-3000 mg, more preferably 100pg-2000mg, more preferably 1 mg-1000 mg, more preferably 10

mg-500 mg, more preferably 100 mg-300mg (in terms of the total amount of ginsenoside Rk2, ginsenoside CK and ginsenoside PPT (or ginsenoside Rk2 and ginsenoside PPT, or ginsenoside CK

and ginsenoside PPT)) per day . The above administration amount can usually be administered in 1-3

doses over the course of a day.

Example

The examples given below are intended to facilitate understanding of the present application, and do not limit the scope of the claims of the present application in any way.

Example 1 Inhibitory effects of rare ginsenoside Rk2/CK/PPT compositions on the

proliferation of three different tumor cells Appropriate amounts of ginsenoside Rk2, ginsenoside CK and ginsenoside PPT were

respectively weighed to prepare aqueous solutions with a final concentration of 100 [g/mL, and the

solutions were referred to as Rk2, CK, and PPT in sequence. Ginsenoside Rk2, ginsenoside CK and ginsenoside PPT were mixed according to the weight ratios of 1: 1: 1, 1: 1.5: 1.5, 1: 2: 2, 1: 0.5: 0.5

and 1: 3: 3 respectively, to prepare aqueous solutions comprising nsenoside Rk2, ginsenoside CK and

ginsenoside PPT with a final concentration of 100 [g/mL. The aqueous solutions were referred to as composition 1, composition 2, composition 3, composition 4 and composition 5 in sequence.

Ginsenoside Rk2 and ginsenoside CK were mixed according to the weight ratios of 1: 1, 1: 0.5, and 1:

3 to prepare aqueous solutions comprising ginsenoside Rk2 and ginsenoside CK with a final concentration of 100 [g/mL, and the solutions were referred to as composition 6, composition 7 and

composition 8. Ginsenoside Rk2 and ginsenoside PPT were mixed according to the weight ratios of 1:

1, 1: 0.5, and 1: 3 to prepare aqueous solutions comprising ginsenoside Rk2 and ginsenoside PPT with a final concentration of 100 [g/mL, which were referred to as composition 9, composition 10 and composition 11. Ginsenoside CK and ginsenoside PPT were mixed according to the weight ratios of 1: 1, 1: 0.5, 1: 3 to prepare aqueous solutions comprising ginsenoside CK and ginsenoside PPT with a final concentration of 100 [g/mL, which were referred to as composition 12, composition 13 and composition 14.

Lung cancer cells, breast cancer cells, and colon cancer cells were used as experimental cell

lines. The three types of cancer cells were inoculated into sterile 96-well plates in turn. After culturing with RPMI-1640 medium for 48 hours, the above 17 kinds of saponin aqueous solutions

were added respectively, 100 L for each well. The control group was added with an equal volume of

RPMI-1640 medium, and 5 replicate wells were set for each concentration. After culturing for 48 hours, 5 g/mL MTT was added to each well, and the culture was continued for 4 hours. 150 L of

DMSO was added and shaken slowly for 10 minutes to dissolve the formed blue-purple crystal

Formazan. The absorbance A value corresponding to each well was detected at 570 nm, and the cell inhibition rate was calculated using the following formula:

Inhibition rate = (1-average OD value of the administration group/average OD value of the

control group) x 100% The two-drug combination index CI of the composition was calculated simultaneously (see,

David H. Kern, Carol R. Morgan, and Susanne U. Hildebrand-Zanki. In vitro and in vivo interaction

between cisplatin and topotecan in ovarian carcinoma systems [J]. Cancer Research,1988,48; and Li Xingqi, the inhibitory effect of lidamycin on glioma and the synergistic effect of combined

temozolomide [D], China Union Medical College, 2009.):

CI= AB/(AxB) Wherein T is the OD value of the experimental group, C is the OD value of the control group,

AB is the T/C value of the saponin combination group, and A and B are the T/C values of the groups

applied with a single saponin. When CI < 1, the two had a synergistic effect, and when CI< 0.7, the synergistic effect was very significant.

The three-drug combination index CI of the composition:

CI= ABC/(AxBxC) Wherein T is the OD value of the experimental group, C is the OD value of the control group,

ABC is the T/C value of the saponin combination group, and A, B, and C are the T/C values of the

groups applied with a single saponin. When CI < 1, the three had a synergistic effect, and when CI < 0.7, the synergistic effect was very significant. Inhibitory effects and combination indices of the above Rk2, CK, PPT and compositions 1-14 on the three tumor cells are shown in Tables 1-4.

From Tables 1-4, it can be seen that ginsenoside Rk2, ginsenoside CK and ginsenoside PPT all showed certain inhibitory effects on the three types of tumor cells. The combination of ginsenoside Rk2, ginsenoside CK and ginsenoside PPT could effectively improve the inhibition rate on three

tumor cells, and showed statistical significance compared with the control (P<0.05). By changing the

specific weight ratio of ginsenoside Rk2, ginsenoside CK and ginsenoside PPT in the composition (compositions 1-3), the anti-tumor effect was more obvious, and the combination indices were all

less than 1. However, either composition 4 or composition 5 did not show a synergistic effect (CI>1).

When any two of ginsenoside Rk2, ginsenoside CK and ginsenoside PPT were combined in a specific ratio, the inhibition rate was significantly increased, and the combination indices were less

than 1 (the results are shown in Tables 3-4). In addition, composition 1 exhibited a more significant

inhibitory effect than the other compositions. The above results indicated that the composition of the present application exerted a synergistic effect.

Table 1 Inhibitory effects of Rk2, CK, PPT and compositions 1-5 on 3 types of tumor cells

Composition Composition Composition Composition Composition Name Control Rk2 CK PPT 1 2 3 4 5

ung cancer 11 inhibition 0.03% 54.92%** 53.31%** 58.76%** 95.20%** 94.70%** 93.59%** 87.32%** 86.51%** rate (%)

east cancer 11 inhibition 0.02% 51.56%** 60.47%** 51.22%** 95.00%** 93.14%** 93.54%** 86.27%** 86.29%** rate (%)

olon cancer 11 inhibition 0.04% 52.51%** 55.74%** 56.23%** 94.65%** 94.30%** 93.42%** 87.95%** 86.07%** rate (%)

The value in the table is the inhibition rate, the unit is %

Compared with the blank control group: *P<0.05, **P<0.01.

Table 2 Combination indices (CI indices) of compositions 1-5 on 3 types of tumor cells Name Lung cancer cells Breast cancer cells Colon cancer cells Control OD value 1.000 0.900 0.800 OD value 0.451 0.436 0.380 Rk2 T/C value 0.451 0.484 0.475 OD value 0.467 0.356 0.354 CK T/C value 0.467 0.395 0.443

OD value 0.412 0.439 0.350 PPT T/C value 0.412 0.488 0.438 OD value 0.048 0.045 0.043 Composition 1 T/C value 0.048 0.050 0.054 CI index 0.55 0.54 0.58 OD value 0.053 0.062 0.046 Composition 2 T/C value 0.053 0.069 0.057 CI index 0.61 0.73 0.62 OD value 0.064 0.058 0.053 Composition 3 T/C value 0.064 0.065 0.066 CI index 0.74 0.69 0.72 OD value 0.127 0.124 0.096 Composition 4 T/C value 0.127 0.137 0.121 CIindex 1.46 1.47 1.31 OD value 0.135 0.123 0.111 Composition 5 T/C value 0.135 0.137 0.139 CI index 1.55 1.47 1.51

Table 3 Inhibitory effects of Rk2, CK, PPT and compositions 6-14 on 3 types of tumor cells Breast cancer cells Lung cancer cells inhibition rate Colon cancer cells Name inhibition rate(%) inhibition rate(%)

Control 0.03% 0.02% 0.04% Rk2 54.92%** 51.56%** 52.51%** CK 53.31%** 60.47%** 55.74%** PPT 58.76%** 51.22%** 56.23%** Composition 6 87.40%** 87.93%** 86.99%** Composition 7 70.55%** 72.24%** 71.87%** Composition 8 69.76%** 73.86%** 70.62%** Composition 9 88.33%** 85.86%** 87.75%** Composition 10 72.41%** 68.64%** 69.74%** Composition 11 73.75%** 67.00%** 70.75%** Composition 12 87.94%** 88.12%** 88.05%** Composition 13 73.43%** 72.55%** 73.74%** Composition 14 72.74%** 70.45%** 72.76%**

The value in the table is the inhibition rate, the unit is %

Compared with the blank control group: *P<0.05, **P<0.01.

Table 4 Combination indices (CI indices) of compositions 6-14 on 3 types of tumor cells Name Lung cancer cells Breast cancer cells Colon cancer cells Control OD value 1.000 0.900 0.800 OD value 0.451 0.436 0.380 Rk2 T/C value 0.451 0.484 0.475 OD value 0.467 0.356 0.354 CK T/C value 0.467 0.395 0.443 OD value 0.412 0.439 0.350 PPT T/C value 0.412 0.488 0.438 OD value 0.126 0.109 0.104 Composition 6 T/C value 0.126 0.121 0.130 CI index 0.60 0.63 0.62 OD value 0.295 0.250 0.225 Composition 7 T/C value 0.295 0.278 0.281 CI index 1.40 1.45 1.34 OD value 0.302 0.235 0.235 Composition 8 T/C value 0.302 0.261 0.294 CI index 1.44 1.37 1.40 OD value 0.117 0.127 0.098 Composition 9 T/C value 0.117 0.141 0.123 CI index 0.63 0.60 0.59 OD value 0.276 0.282 0.242 Composition 10 T/C value 0.276 0.314 0.303 CI index 1.48 1.33 1.46 OD value 0.263 0.297 0.234 Composition 11 T/C value 0.263 0.330 0.293 CI index 1.41 1.40 1.41 OD value 0.121 0.107 0.096 Composition 12 T/C value 0.121 0.119 0.120 CI index 0.63 0.62 0.62 OD value 0.266 0.247 0.210 Composition 13 T/C value 0.266 0.275 0.263 CI index 1.38 1.42 1.36 Composition 14 OD value 0.273 0.266 0.218

T/C value 0.273 0.296 0.272 CI index 1.42 1.53 1.41

Example 2 Effects of rare ginsenoside Rk2/CK/PPT composition 1, composition 6, composition 9 and composition 12 on the distribution of colon cancer cell cycle

Colon cancer cells were seeded in sterile 6-well plates, and after culturing for 24 hours, ginsenoside Rk2, ginsenoside CK, ginsenoside PPT, composition 1, composition 6, composition 9

and composition 12 were added at a concentration of 100 [g/mL. The control group was added with

an equal volume of RPMI-1640 medium, and 5 replicate wells were set for each concentration. After continuing to culture for 48 hours, ice-cold PBS was added to wash 2-3 times, 75% ice ethanol was

added to adjust the cells to a suitable density, and the cells were placed in a refrigerator at 4°C

overnight. Then the cells were added with ice-cold PBS to wash 2-3 times. An appropriate amount of PI solution was added dropwise, and the cells were incubated in the dark for 30 minutes. Flow

cytometry was used to detect the cell cycle distribution.

Figure 1 is a graph showing the experimental results of the effects of rare ginsenoside composition 1, composition 6, composition 9 and composition 12 on the distribution of colon cancer

cell cycle detected by flow cytometry. After the colon cancer cells were treated with ginsenoside Rk2,

ginsenoside CK, ginsenoside PPT, composition 1, composition 6, composition 9 and composition 12 respectively, compared with the control, the proportion of cells in G phase increased significantly, the proportion of cells in S phase decreased, and the proportion of cells in G2 phase did not change

significantly. Ginsenoside Rk2, ginsenoside CK, ginsenoside PPT, composition 1, composition 6, composition 9, and composition 12 increased the proportion of colon cancer cells in GI phase to

49.97%, 48.09%, 49.39%, 68.23%, 55.98%, 55.68%, and 56.10%, respectively. Compared with the

control group, they all showed statistical differences (P<0.01); the above data showed that after treatment with ginsenoside Rk2, ginsenoside CK and ginsenoside PPT, colon cancer cells were

blocked in the GI phase, and the composition combined in a specific ratio showed an obvious

blocking effect. Table 5 Effects of composition 1, composition 6, composition 9, and composition 12 on the

blocking of colon cancer cell cycle distribution Group G1%

Control 30.98%

Rk2 49.97%**

CK 48.09%**

PPT 49.39%**

Composition 1 68.23%**

Composition 6 55.98%**

Composition 9 55.68%**

Composition 12 56.10%**

Compared with the blank control group: *P<0.05, **P<0.01.

Example 3 Apoptosis-inducing effects of rare ginsenoside Rk2/CK/PPT composition 1,

composition 6, composition 9 and composition 12 on colon cancer cells Colon cancer cells were seeded in sterile 6-well plates, and after culturing for 24 hours, ginsenoside Rk2, ginsenoside CK, ginsenoside PPT, composition 1, composition 6, composition 9

an equal volume of RPMI-1640 medium, and 5 replicate wells were set for each concentration. Continuing to culture for 48 hours, the cells were added with ice-cold PBS to wash 2-3 times. 1x

Binding Buffer was added and pipetted evenly, an appropriate amount of AV/PI mixed dye solution

was added dropwise, and the cells were incubated in the dark for 15 minutes. Flow cytometry was used to detect cell apoptosis, and the two-drug combination index CI of the composition was calculated:

CI = (1-AxB)/(1-AB) Wherein T is the cell survival fraction of the experimental group, C is the cell survival fraction

of the control group, AB is the T/C value of the saponin combination group, A and B are the survival

fraction T/C values of the groups applied with a single saponin. When CI < 1, the two have a synergistic effect, and when CI < 0.7, the synergistic effect is very significant.

The three-drug combination index CI of the composition:

CI= (1-AxBxC)/(1-ABC) Wherein T is the cell survival fraction of the experimental group, C is the cell survival fraction

of the control group, ABC is the T/C value of the saponin combination group, and A, B, and C are

the survival fraction T/C values of the groups applied with a single saponin. When CI < 1, the three have a synergistic effect, and when CI < 0.7, the synergistic effect is very significant.

Figure 2 is a graph showing the experimental results of the apoptosis of colon cancer cells

induced by rare ginsenoside composition 1, composition 6, composition 9 and composition 12

detected by flow cytometry. A cell histogram composed of four quadrants is obtained, wherein the region Q Irepresents the cells with mechanical damage during the experimental operation, the region

Q2 represents the cells with late apoptosis, the region Q3 represents the cells with normal function

and morphology, and the region Q4 represents the cells with early apoptosis. Ginsenoside Rk2, ginsenoside CK and ginsenoside PPT could induce certain apoptosis of colon cancer cells, and the

rare ginsenoside composition 1, composition 6, composition 9 and composition 12 of the present

application significantly increased the percentage of apoptotic cells in colon cancer cells. The early apoptosis rates of colon cancer cells treated with ginsenoside Rk2, ginsenoside CK and ginsenoside

PPT were 11.32%, 10.01% and 12.73% respectively; the early apoptosis rates of colon cancer cells

treated with composition 1, composition 6, composition 9 and composition 12 of the present application were 40.38%, 24.34%, 27.67% and 25.89%, respectively. Compared with the control

group, they had significant statistical differences (P<0.01), and the combination indices of

composition 1, composition 6, composition 9 and composition 12 in colon cancer cell apoptosis induction were 0.59, 0.67, 0.68, and 0.68 (the results were shown in Table 6), showing a synergistic

effect. The above results showed that ginsenoside Rk2/CK/PPT composition promoted the

accelerated apoptosis of colon cancer cells, thereby playing a role in the treatment of colon cancer.

Table 6 Combination indices (CI indices) of composition 1, composition 6, composition 9, and

composition 12 in colon cancer cell apoptosis induction Group Q4% T/C value CI index Control 3.43% -- -

Rk2 11.32%** 0.918 -

CK 10.01%** 0.932 -

PPT 12.73%** 0.904 - Composition 1 40.38%** 0.617 0.59 Composition 6 24.34%** 0.783 0.67 Composition 9 27.67%** 0.749 0.68 Composition 12 25.89%** 0.767 0.68

Compared with the blank control group: *P<0.05, **P<0.01.

Example 4 Effect of composition 1 on the expression of Bcl-2 family proteins, Caspase-3 and

PARP proteins in colon cancer cells

Colon cancer cells were seeded into 6-well plates, and ginsenoside Rk2, ginsenoside CK, ginsenoside PPT and composition 1 were added at a concentration of 100 [g/mL. After culturing for

48 hours, trypsin was added to digest the cells, and an appropriate amount of RIPA lysis solution was

added. After 30 minutes, target samples were obtained by centrifuging. Then, after gel preparation, SDS-PAGE electrophoresis, membrane transfer, blocking and incubation, and DAB color

development, the expression map of apoptosis-related proteins was obtained.

Figure 3 shows the expression results of apoptosis related proteins in colon cancer cell induced by ginsenoside Rk2, ginsenoside CK, ginsenoside PPT and composition 1 detected by Western

blotting. Compared with the control, after treatment with ginsenoside Rk2, ginsenoside CK and

ginsenoside PPT, the expressions of Caspase-3, PARP and Bcl-2 proteins were decreased; After treatment with composition 1, the changes in the expression levels of the above three proteins were

more obvious. It can be seen that composition 1 finally induces apoptosis through Caspase-3

activation and Bcl-2 protein downregulation.

Example 5 Inhibitory effects of rare ginsenoside Rk2/CK/PPT composition 1, composition 6,

composition 9 and composition 12 on the colon cancer xenograft model Male BALB/c nude mice were fed at room temperature, and after adapted to the environment,

they were inoculated with colon cancer cells, and each nude mouse was injected with 3-4x106 cells.

Seven days after inoculation, the subcutaneous left armpit of nude mice was observed, and it was found that nodules at the size of rice grains were formed at the inoculation sites of nude mice. The

feeding was continued, and when the tumor size exceeded 100 mm 3, the nude mice with large

differences in tumor size and body weight were excluded, and the remaining tumor-bearing nude mice were randomly divided into groups. In this experiment, all tumor-bearing nude mice were

divided into the following 4 groups: (1) model group; (2) 10 mg/kg Rk2 group; (3) 10 mg/kg CK

group; (4) 10 mg/kg PPT group; (4) composition 1 group (10 mg/kg); (5) composition 6 group (10 mg/kg); (6) composition 9 group (10 mg/kg); and (7) composition 12 group (10 mg/kg). The model

group was given a mixture of polyethylene glycol and water (volume ratio of 1:1). The other groups

were given intragastric administration according to the body weight of tumor-bearing nude mice, once a day. The body weight, tumor volume and dietary changes of tumor-bearing nude mice were measured every 5 days, and the records were observed in time. After 30 days, the administration was stopped, all mice were sacrificed, and tumor masses were excised, weighed, recorded and collected one by one. At the same time, the CI of the two-drug or three-drug combination indices on the 30th day was calculated: Two-drug combination CI= AB/(AxB) Wherein T is the tumor weight in the experimental group, C is the tumor weight in the control group, AB is the T/C value of the two saponin-combination group, A and B are the survival fraction T/C values of the groups applied with a single saponin. When CI < 1, the two have a synergistic effect, and when CI < 0.7, the synergistic effect is very significant. Three-drug combination CI=ABC/(AxBxC) Wherein T is the tumor weight of the experimental group, C is the tumor weight of the control group, ABC is the T/C value of the saponin combination group, and A, B, and C are the T/C values of the groups applied with a single saponin. When CI < 1, the three have a synergistic effect, and when CI < 0.7, the synergistic effect is very significant. It can be seen from Figure 4A and Tables 7-8 that compared with the model group, the tumor inhibition rates of transplanted tumors in colon cancer nude mice of ginsenoside Rk2 group, ginsenoside CK group and ginsenoside PPT group were 27.15%, 33.18% and 34.05% respectively on the 30th day of administration, indicating that ginsenoside Rk2 group, ginsenoside CK group and ginsenoside PPT group showed certain inhibitory effects on the colon cancer transplanted tumors in nude mice. In composition 1 group, composition 6 group, composition 9 group and composition 12 group, the tumor weight of tumor-bearing mice could be significantly reduced, and the corresponding tumor inhibition rates were 79.74%, 64.65%, 65.51%, and 67.24%, respectively with a significant effect (P<0.01), and the combination indices were 0.63, 0.73, 0.72, and 0.74, respectively (see Table 7-8 for the results). Compared with the model group, the body weight of the nude mice in the ginsenoside Rk2 group, ginsenoside CK group, ginsenoside PPT group, composition 1 group, composition 6 group, composition 9 group and composition 12 group showed a slow increase trend, indicating low toxicity and high safety (Figure 4B). In conclusion, the rare ginsenoside Rk2/CK/PPT composition could significantly inhibit the growth of the colon cancer transplanted tumor in nude mice, and the inhibition rate was significantly higher than that of the ginsenoside Rk2 group, ginsenoside CK group and ginsenoside PPT group, and showed low toxicity and side effect to nude mice. The rare ginsenoside Rk2/CK/PPT composition provided by the application showed a synergistic effect. Table 7 Inhibitory effects of composition 1, composition 6, composition 9, and composition 12 on the growth of colon cancer tumor-bearing mice Group Dose (mg/kg) Tumor weight (g) Tumor inhibition rate(%) Model group -- 2.32 -

Ginsenoside Rk2 group 10 1.69** 27.15% Ginsenoside CK group 10 1.55** 33.18% Ginsenoside PPT group 10 1.53** 34.05% Composition 1 group 10 0.47** 79.74% Composition 6 group 10 0.82** 64.65% Composition 9 group 10 0.80** 65.51% Composition 12 group 10 0.76** 67.24%

Compared with the model group: *P<0.05, **P<0.01.

Table 8 Combination indices (CI indices) of composition 1, composition 6, composition 9, and

composition 12 on tumor regrowth inhibition in colon cancer tumor-bearing mice Group Tumor weight (g) T/C value CI index Control 2.32 -- -

Rk2 1.69 0.728 -

CK 1.55 0.668 PPT 1.53 0.659 -

Composition 1 0.47 0.203 0.63 Composition 6 0.82 0.353 0.73 Composition 9 0.8 0.345 0.72 Composition 12 0.76 0.328 0.74

Example 6 In vivo safety evaluation of ginsenoside Rk2/CK/PPT composition 1

Male healthy ICR mice were fed in a room temperature environment, with diet freedom, -60% relative humidity, and 12 h day/12 h night. After adapted to the environment, they were fed

for 1 week and fasted for 12 hours. After 12 hours, the mice were randomly divided into 4 groups: (1)

blank group; (2) 40 mg/kg ginsenoside Rk2 group; (3) 40 mg/kg ginsenoside CK group; (4) 40 mg/kg ginsenoside PPT group; and (5) Rk2/CK/PPT composition 1 group. The above 5 groups were all administered by gavage. 6 hours after administration, the fasting was canceled, and the mice were fed normally for 14 days. Changes in drinking water, food intake, body weight, and liver and kidney function of mice were observed. It can be seen from Figure 5 that compared with the blank group, after exposure, the water intake, food intake and body weight of the mice in the ginsenoside Rk2 group, ginsenoside CK group and ginsenoside PPT group did not change significantly; and the water intake, food intake and body weight of the mice in the Rk2/CK/PPT composition 1 group did not change significantly. It can be seen from Figure 6 that, compared with the control group, the liver weight, kidney weight, serum alanine aminotransferase content and serum creatinine content of the mice in ginsenoside Rk2 group, ginsenoside CK group and ginsenoside PPT group did not change significantly; and the liver weight, kidney weight, serum alanine aminotransferase content in and serum creatinine content of the mice in the Rk2/CK/PPT composition 1 group did not significantly change. It can be seen that the Rk2/CK/PPT composition 1 showed certain safety to healthy mice. It should be noted that on the premise that it can be implemented and does not obviously violate the gist of the present application, any technical feature or combination of technical features described in this specification as a constituent part of a technical solution can also be applied to other technical solutions; and on the premise that it can be implemented and does not obviously violate the gist of the present application, the technical features described as constituent parts of different technical solutions can also be combined in any way to form other technical solutions. The present application also includes technical solutions obtained by combining in the above-mentioned cases, and these technical solutions are equivalent to being described in this specification. The present application has been described above through specific embodiments and examples, but those skilled in the art should understand that these are not intended to limit the scope of the present application, which should be determined by the claims.

Industrial Applicability The present application provides a rare ginsenoside pharmaceutical composition comprising rare ginsenoside Rk2, ginsenoside CK and ginsenoside PPT, which is synergistic and has anti-tumor effects.

Claims

1. A rare ginsenoside pharmaceutical composition with anti-tumor effect, comprising therapeutically effective amounts of ginsenoside Rk2, ginsenoside CK and ginsenoside PPT as active

ingredients, and a pharmaceutically acceptable carrier,

wherein in the rare ginsenoside pharmaceutical composition, the weight ratio of ginsenoside Rk2, ginsenoside CK and ginsenoside PPT is: 1: (1 to 2): (1 to 2).

2. The rare ginsenoside pharmaceutical composition according to claim 1, wherein in the rare

ginsenoside pharmaceutical composition, the weight ratio of ginsenoside Rk2, ginsenoside CK and ginsenoside PPT is: 1: (1 to 1.1): (1 to 1.1).

3. The rare ginsenoside pharmaceutical composition according to claim 1, wherein in the rare

ginsenoside pharmaceutical composition, the weight ratio of ginsenoside Rk2, ginsenoside CK and ginsenoside PPT is 1: 1: 1.

4. The rare ginsenoside pharmaceutical composition according to claim 1, wherein the rare

ginsenoside pharmaceutical composition does not comprise other ginsenosides.

5. The rare ginsenoside pharmaceutical composition according to claim 1, wherein the rare

ginsenoside pharmaceutical composition comprises ginsenoside Rk2, ginsenoside CK and

ginsenoside PPT as the only anti-tumor active ingredients.

6. Use of a composition comprising ginsenoside Rk2, ginsenoside CK and ginsenoside PPT in

the preparation of an anti-tumor medicament, wherein in the composition, the weight ratio of

ginsenoside Rk2, ginsenoside CK and ginsenoside PPT is: 1: (1 to 2): (1 to 2); and the anti-tumor medicament comprises ginsenoside Rk2, ginsenoside CK and ginsenoside

PPT as active ingredients.

7. The use according to claim 6, wherein in the composition, the weight ratio of ginsenoside Rk2, ginsenoside CK and ginsenoside PPT is: 1: (1 to 1.1): (1 to 1.1).

8. The use according to claim 6, wherein in the composition, the weight ratio of ginsenoside

Rk2, ginsenoside CK and ginsenoside PPT is 1:1:1. 9. The use according to claim 6, wherein the composition does not comprise other ginsenosides.

10. The use according to claim 6, wherein the composition comprises ginsenoside Rk2,

ginsenoside CK and ginsenoside PPT as the only anti-tumor active ingredients.