阅读说明：本技术 路路通内酯在治疗癌症中的应用 (Application of liquidambar formosana hance lactone in treating cancer ) 是由 屈祎 闫蓉 柯细松 张卫东 黄飘 于 2020-09-23 设计创作，主要内容包括：本发明发现中药路路通中得到的式I所示化合物对Wnt信号通路表现出靶向抑制作用。因此,式I所示化合物能够开发成为全新的Wnt抑制剂以及治疗癌症,特别是大肠癌的药物,从而为开发中药来源的抗肿瘤药物开辟了新的方向。(The invention discovers that the compound shown in the formula I obtained from the traditional Chinese medicine fructus liquidambaris shows a targeted inhibition effect on the Wnt signal path. Therefore, the compound shown in the formula I can be developed into a brand-new Wnt inhibitor and a medicament for treating cancers, particularly colorectal cancers, so that a new direction is opened for developing antitumor medicaments from traditional Chinese medicines.)

1. The application of the liquidambar formosana hance active extract, or the compound shown in the formula I or the pharmaceutically acceptable salt thereof in preparing the Wnt/beta-catenin signal channel inhibitor is as follows:

2. the use according to claim 1, wherein the inhibitor of the Wnt/β -catenin signaling pathway is a therapeutic drug, food or health product for diseases related to the Wnt/β -catenin signaling pathway.

3. The use according to claim 2, wherein the Wnt/β -catenin signaling pathway related diseases are cancer, nervous system diseases, bone diseases, skin diseases, cardiovascular diseases, fibrotic diseases and metabolic syndrome.

4. The use of claim 3, wherein the cancer is colorectal cancer, liver cancer, gastric cancer, lung cancer, prostate cancer, pancreatic cancer, breast cancer, ovarian cancer, and osteosarcoma.

5. The use of claim 4, wherein the cancer is colorectal cancer.

6. A pharmaceutical composition comprising a compound of formula I or a pharmaceutically acceptable salt thereof or a Liquidambar formosana Hance active extract containing an effective amount, in particular a therapeutically effective amount, of a compound of formula I, and optionally a pharmaceutically acceptable excipient,

7. a health product or food composition comprising a compound of formula I or a pharmaceutically acceptable salt thereof or a Liquidambar formosana Hance active extract containing an effective, in particular therapeutically effective amount of a compound of formula I, and optionally a pharmaceutically acceptable excipient,

8. a pharmaceutical composition or health product or food composition containing fructus Liquidambaris active extract or compound shown in formula I or pharmaceutically acceptable salt thereof for treating or adjunctively treating Wnt/beta-catenin signaling pathway related diseases,

9. the fructus liquidambaris active extract or the compound shown in the formula I or the pharmaceutically acceptable salt thereof is used as a Wnt/beta-catenin signal channel inhibitor.

10. The fructus liquidambaris active extract or the compound shown in the formula I or the pharmaceutically acceptable salt thereof is used for treating Wnt/beta-catenin signal channel related diseases.

Technical Field

The invention relates to the field of medicines. In particular, the invention relates to the application of liquidambar formosana lactone in treating cancers, particularly colorectal cancers.

Background

Colorectal cancer is located at the 3 rd site of malignant tumors that are common worldwide, and the mortality rate is located at the 4 th site. In recent 40 years, colorectal cancer has become the 3 rd common malignant tumor in China, and accounts for 10.30% of all malignant tumors. The high incidence of colorectal cancer causes a heavy medical and economic burden. The main treatment methods of colorectal cancer are surgery, radiotherapy by means of instruments, and adjuvant therapy with chemicals, however, even if the three methods are combined, the recovery of patients cannot be guaranteed, and postoperative recurrence is easy. And related research shows that the five-year survival rate of colon cancer patients after radical treatment in some developed countries is less than 60%.

Therefore, there is an urgent need in the art to develop novel targeted therapeutic drugs that can be used for colorectal cancer.

Disclosure of Invention

The invention aims to provide a novel targeted therapeutic drug for colorectal cancer, thereby providing a method for treating colorectal cancer with slight adverse reaction.

In a first aspect, the invention provides the use of an active extract of fructus liquidambaris, or a compound shown in formula I or a pharmaceutically acceptable salt thereof, in the preparation of a Wnt/beta-catenin signaling pathway inhibitor:

in a preferred embodiment, the fructus liquidambaris extract is an extract containing an effective amount, in particular a therapeutically effective amount, of a compound represented by formula I.

In a preferred embodiment, the active extract of fructus liquidambaris is fructus liquidambaris extract, fructus liquidambaris fine powder, fructus liquidambaris extract, fructus liquidambaris concentrated juice and fructus liquidambaris volatile oil.

In a specific embodiment, the Wnt/beta-catenin signal pathway inhibitor is a therapeutic drug or food or health product for diseases related to the Wnt/beta-catenin signal pathway.

In a preferred embodiment, the Wnt/β -catenin signaling pathway-related disease is a disease in which the Wnt/β -catenin signaling pathway is overexpressed.

In a specific embodiment, the Wnt/β -catenin signaling pathway related diseases are cancer, nervous system diseases, bone diseases, skin diseases, cardiovascular diseases, fibrotic diseases, metabolic syndrome, and the like.

In specific embodiments, the cancer is colorectal cancer, liver cancer, gastric cancer, lung cancer, prostate cancer, pancreatic cancer, breast cancer, ovarian cancer, and osteosarcoma.

In a specific embodiment, the cancer is colorectal cancer.

In a preferred embodiment, the cancer is an in situ cancer or a metastatic tumor.

In a second aspect, the present invention provides a pharmaceutical composition comprising a compound of formula I or a pharmaceutically acceptable salt thereof, or a Liquidambar formosana Hance active extract containing an effective amount, in particular a therapeutically effective amount, of a compound of formula I, and optionally a pharmaceutically acceptable excipient,

in a preferred embodiment, the pharmaceutical composition is a Wnt/beta-catenin signal pathway inhibitor.

In a preferred embodiment, the Wnt/beta-catenin signal pathway inhibitor is a therapeutic drug for diseases related to the Wnt/beta-catenin signal pathway.

In a preferred embodiment, the Wnt/beta-catenin signal pathway related diseases are diseases in which the Wnt/beta-catenin signal pathway is over-expressed; including but not limited to cancer, neurological disorders, bone disorders, skin disorders, cardiovascular disorders, fibrotic disorders and metabolic syndrome.

In a preferred embodiment, the cancer is colorectal cancer, liver cancer, stomach cancer, lung cancer, prostate cancer, pancreatic cancer, breast cancer, ovarian cancer, and osteosarcoma; colorectal cancer is preferred.

In a third aspect, the present invention provides a nutraceutical or food composition comprising a compound of formula I or a pharmaceutically acceptable salt thereof or a Lipuidambaris active extract comprising an effective amount, in particular a therapeutically effective amount, of a compound of formula I, and optionally a pharmaceutically acceptable excipient,

in a preferred embodiment, the active extract of fructus liquidambaris is fructus liquidambaris extract, fructus liquidambaris fine powder, fructus liquidambaris extract, fructus liquidambaris concentrated juice and fructus liquidambaris volatile oil.

In a fourth aspect, the invention provides a pharmaceutical composition or health product or food composition containing fructus liquidambaris active extract or a compound shown in formula I or pharmaceutically acceptable salt thereof for treating or adjunctively treating Wnt/beta-catenin signaling pathway related diseases,

in a fifth aspect, the invention provides a liquidambar formosana hance active extract or a compound shown in a formula I or a pharmaceutically acceptable salt thereof, which is used as a Wnt/beta-catenin signal channel inhibitor.

In a preferred embodiment, the fructus liquidambaris extract is an extract containing an effective amount, in particular a therapeutically effective amount, of a compound represented by formula I.

In a preferred embodiment, the active extract of fructus liquidambaris is fructus liquidambaris extract, fructus liquidambaris fine powder, fructus liquidambaris extract, fructus liquidambaris concentrated juice and fructus liquidambaris volatile oil.

In a preferred embodiment, the Wnt/beta-catenin signal pathway inhibitor is a therapeutic drug or food or health product for diseases related to the Wnt/beta-catenin signal pathway.

In a preferred embodiment, the Wnt/beta-catenin signal pathway related diseases are diseases in which the Wnt/beta-catenin signal pathway is over-expressed; including but not limited to cancer, neurological disorders, bone disorders, skin disorders, cardiovascular disorders, fibrotic disorders and metabolic syndrome.

In a preferred embodiment, the cancer is colorectal cancer, liver cancer, stomach cancer, lung cancer, prostate cancer, pancreatic cancer, breast cancer, ovarian cancer, and osteosarcoma; colorectal cancer is preferred.

In a preferred embodiment, the cancer is carcinoma in situ or a metastatic carcinoma.

In a sixth aspect, the invention provides a liquidambar formosana hance active extract or a compound shown in a formula I or a pharmaceutically acceptable salt thereof, which is used for treating Wnt/beta-catenin signaling pathway related diseases.

In a preferred embodiment, the fructus liquidambaris extract is an extract containing an effective amount, in particular a therapeutically effective amount, of a compound represented by formula I.

In a preferred embodiment, the active extract of fructus liquidambaris is fructus liquidambaris extract, fructus liquidambaris fine powder, fructus liquidambaris extract, fructus liquidambaris concentrated juice and fructus liquidambaris volatile oil.

In a preferred embodiment, the Wnt/beta-catenin signal pathway inhibitor is a therapeutic drug or food or health product for diseases related to the Wnt/beta-catenin signal pathway.

In a preferred embodiment, the Wnt/beta-catenin signal pathway related diseases are diseases in which the Wnt/beta-catenin signal pathway is over-expressed; including but not limited to cancer, neurological disorders, bone disorders, skin disorders, cardiovascular disorders, fibrotic disorders and metabolic syndrome.

In a preferred embodiment, the cancer is colorectal cancer, liver cancer, stomach cancer, lung cancer, prostate cancer, pancreatic cancer, breast cancer, ovarian cancer, and osteosarcoma; colorectal cancer is preferred.

In a preferred embodiment, the cancer is carcinoma in situ or a metastatic carcinoma.

In a seventh aspect, the present invention provides a method for treating diseases related to Wnt/β -catenin signaling pathway, which comprises administering a therapeutically effective amount of a compound represented by formula I or a pharmaceutically acceptable salt thereof or an active extract of fructus liquidambaris to a patient in need of treatment of the diseases.

In a preferred embodiment, the fructus liquidambaris extract is an extract containing an effective amount, in particular a therapeutically effective amount, of a compound represented by formula I.

In a preferred embodiment, the active extract of fructus liquidambaris is fructus liquidambaris extract, fructus liquidambaris fine powder, fructus liquidambaris extract, fructus liquidambaris concentrated juice and fructus liquidambaris volatile oil.

In a preferred embodiment, the Wnt/beta-catenin signal pathway related diseases are diseases in which the Wnt/beta-catenin signal pathway is over-expressed; including but not limited to cancer, neurological disorders, bone disorders, skin disorders, cardiovascular disorders, fibrotic disorders and metabolic syndrome.

In a preferred embodiment, the cancer is colorectal cancer, liver cancer, stomach cancer, lung cancer, prostate cancer, pancreatic cancer, breast cancer, ovarian cancer, and osteosarcoma; colorectal cancer is preferred.

In a preferred embodiment, the cancer is carcinoma in situ or a metastatic carcinoma.

It is to be understood that within the scope of the present invention, the above-described features of the present invention and those specifically described below (e.g., in the examples) may be combined with each other to form new or preferred embodiments. Not to be reiterated herein, but to the extent of space.

Drawings

FIG. 1 shows the effect of compounds of formula I on the formation of clones of 2 colorectal cancer cells, 3 non-small cell lung cancer cells, 4 pancreatic cancer cells, prostate cancer cells, stomach cancer cells, ovarian cancer cells, breast cancer cells, liver cancer cells, and osteosarcoma cells.

FIG. 2 shows the effect of a compound of formula I on subcutaneous transplants of colorectal cancer cells (HCT 116); wherein FIG. 2A shows the inhibition of tumor proliferation by a compound of formula I; fig. 2B shows 14 days after dosing. Effect of the compound of formula I on tumor volume; FIG. 2C shows a photograph of a transplanted tumor after its material is taken; FIG. 2D shows the inhibition of the tumor weight of subcutaneous xenograft tumor of nude mice by the compound of formula I.

FIG. 3 shows that the compound shown in formula I has the function of inhibiting the Wnt/beta-catenin pathway; wherein, FIG. 3A shows that the compound (10, 20. mu.M) shown in formula I can inhibit the transcriptional activity mediated by beta-catenin in cells; FIG. 3B shows the dosing groups and results of zebrafish embryo "eye loss" experiments; FIG. 3C shows the results of 24 hours treatment with Wnt/β -catenin pathway activator (6BIO) in zebrafish embryo 6hpf while administering the compound of formula I (10 μ M) and the same volume of DMSO; FIG. 3D shows the results of administering Wnt/β -catenin pathway activator (6BIO) at 6hpf in zebrafish embryos for 66 hours with the compound of formula I (10 μ M) and the same volume of DMSO (magnification: 40).

Detailed Description

The inventors have conducted extensive and intensive studies and unexpectedly found that the compound represented by formula I exhibits a targeted inhibitory effect on colorectal cancer and Wnt signaling pathway, and thus can be developed into a novel therapeutic drug for colorectal cancer and a novel Wnt inhibitor. The present invention has been completed based on this finding.

Definition of terms

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the disclosed invention belongs. For the purpose of facilitating an understanding of the present invention, the following definitions and explanations will be made with respect to the terms involved in the present invention, but the scope of the present invention is not limited to these specific definitions.

Wnt/beta-catenin signal channel

The Wnt signaling pathway is an evolutionarily well-conserved network of protein action. The canonical Wnt signaling pathway can be briefly summarized as: 1) the Wnt secreted factor binds to receptors (Frizzled family and LRP family) on the cell membrane; 2) the signal leads beta-catenin in cytoplasm to be phosphorylated by a degradation complex consisting of APC, Axin, CK1 alpha and GSK3 beta through a Dishevelled protein, so that the degradation complex can be subjected to ubiquitination degradation mediated by beta-TRCP E3 ligase; 3) beta-catenin aggregates in the cytoplasm and enters the nucleus and TCF/LEF (T cytokine) forms a complex that initiates transcription of a target gene (such as c-myc).

This pathway was first discovered during embryonic development, and aberrant inhibition or activation of the pathway would lead to defects in embryonic development. The Wnt signal pathway is closely related to the events related to tumor invasion and metastasis, such as the migration and adhesion of cancer cells, the degradation of extracellular matrix and the angiogenesis of tumors, and plays an important role in regulating the self-renewal, proliferation and differentiation of tumor stem cells. In addition, it has been shown that the Wnt/β -catenin signaling pathway is associated with chemotherapy resistance. There is now a great deal of evidence that aberrant activation of the Wnt signaling pathway is highly correlated with the development and progression of cancer, particularly colorectal cancer.

Due to the key role of the Wnt/β -catenin signaling pathway in the development of large bowel cancer, numerous research institutes and pharmaceutical companies have been devoted to the development of Wnt inhibitors in the world over the past 20 years. However, no drug targeting the Wnt/β -catenin signaling pathway has been applied clinically so far, and most of the inhibitors are in preclinical studies, with few entering clinical phase 1 or 2. The reason why the development of Wnt inhibitors has progressed slowly is that most inhibitors target the upstream or midstream of the Wnt signaling pathway, but clinical genome sequencing has shown that 81.9% of colorectal cancer tissues have inactivating mutations of the APC gene and activating mutations of 4.7% of the β -catenin gene, which theoretically would result in hindered degradation and abnormal accumulation of β -catenin, and that the Wnt protein targeted by Porcupine inhibitors or Axin targeted by Tankyrase inhibitors precede APC on the pathway, thus having no regulatory effect on these abnormally accumulated β -catenin.

Diseases related to the Wnt/beta-catenin signal pathway comprise cancer, nervous system diseases, bone diseases, skin diseases, cardiovascular diseases, fibrosis diseases, metabolic syndrome and the like; cancers involving the Wnt/β -catenin signaling pathway include, but are not limited to, colorectal cancer, lung cancer, prostate cancer, pancreatic cancer, gastric cancer, liver cancer, breast cancer, ovarian cancer, and osteosarcoma; in particular, large bowel cancer.

The compounds of the present invention and pharmaceutically acceptable salts thereof

As used herein, the terms "compound of the present invention", "compound of formula I" and "liquidambar lactone" have the same meaning and refer to a compound having the structure shown in formula I

The compounds of the invention may be obtained from a variety of sources, for example, may be isolated from traditional Chinese medicine sweetgum fruit. The fructus Lipuidambaris is dry mature fruit of Liquidambar formosana Hance of Hamamelidaceae, and has effects of dispelling pathogenic wind, removing dampness, dredging channels and promoting urination. The compound of the formula I is also called liquidambar formosana hance lactone, and is a pentacyclic triterpene lactone compound separated from liquidambar formosana hance, wherein the molecular formula is C30H44O4, CAS number: 185051-75-6.

On the basis of the compounds of the present invention, those skilled in the art can formulate them into pharmaceutically acceptable salts. For example, the compounds of the present invention may be reacted with inorganic or organic acids to form conventional pharmaceutically acceptable salts. The inorganic acids include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, sulfamic acid, phosphoric acid and the like, and the organic acids include citric acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, benzenesulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, naphthalenesulfonic acid, ethanesulfonic acid, naphthalenedisulfonic acid, maleic acid, malic acid, malonic acid, fumaric acid, succinic acid, propionic acid, oxalic acid, trifluoroacetic acid, stearic acid, pamoic acid, hydroxymaleic acid, phenylacetic acid, benzoic acid, salicylic acid, glutamic acid, ascorbic acid, p-aminobenzenesulfonic acid, 2-acetoxybenzoic acid, isethionic acid and the like; or reacting a compound of the invention with an inorganic base to form a sodium, potassium, calcium, aluminum or ammonium salt; or with an organic base to form a methylamine salt, an ethylamine salt or an ethanolamine salt.

Fructus Liquidambaris active extract

The fructus liquidambaris active extract is a product obtained by taking fructus liquidambaris as a raw material and directionally obtaining and concentrating the fructus liquidambaris lactone in the fructus liquidambaris through extraction and separation processes (such as a decoction method, a reflux method, an impregnation method, a percolation method and the like) of physics and chemistry and the like. In a specific embodiment, the "fructus liquidambaris active extract" as described herein refers to an extract containing an effective amount, in particular a therapeutically effective amount, of a compound of formula I. Thus, the herein described process for obtaining "active extract of sweetgum fruit" comprises the step of enriching or concentrating the sweetgum fruit lactone.

The "fructus Liquidambaris active extract" described herein may take any conventional form known to those skilled in the art, including, but not limited to, fructus Liquidambaris extract, fructus Liquidambaris fine powder, fructus Liquidambaris extract, fructus Liquidambaris concentrated juice, fructus Liquidambaris volatile oil.

The "therapeutically effective amount" as used herein is the same as that conventionally understood by those skilled in the art; in particular, the content of the compound shown in the formula I can play a role in effectively treating certain diseases, such as colorectal cancer. It will be understood by those skilled in the art that the treatment described herein also includes ameliorating effects on the reduction, alleviation, etc. of symptoms of the disease.

Pharmaceutical composition or food or health product composition and corresponding dosage form

On the basis of the compound, namely the liquidambar formosana hance lactone, the invention further provides a pharmaceutical composition or a food or health-care product composition containing the compound or the liquidambar formosana hance active extract as an active ingredient. The pharmaceutical composition comprises the compound or the liquidambar formosana hance active extract and pharmaceutically acceptable excipients, such as pharmaceutically acceptable solid or liquid excipients and/or auxiliary materials. The food or health-care product composition comprises the compound or the active extract of the sweetgum fruit and solid or liquid excipient and/or auxiliary material which is acceptable in health-care products or food science.

The content of the compound of the present invention as an active ingredient in the pharmaceutical composition or food or nutraceutical composition can be autonomously determined by those skilled in the art according to actual needs. For example, the compounds of the present invention are typically present in the pharmaceutical or nutraceutical composition in an amount of 0.1 to 99%.

Based on the teachings of the present invention, the pharmaceutical composition or food or nutraceutical composition of the present invention can be prepared into corresponding dosage forms suitable for administration to a subject or patient in need thereof, such as a human or an animal, using conventional methods by those skilled in the art.

The compounds or pharmaceutical compositions of the present invention may be administered in unit dosage form. The administration route can be gastrointestinal or parenteral, such as oral, intravenous, intramuscular, subcutaneous, nasal, oral mucosal, ocular, pulmonary and respiratory, dermal, vaginal, rectal, etc. Accordingly, the dosage form of the pharmaceutical composition can include, but is not limited to, oral administration preparation, injection administration preparation, and skin mucosa route administration preparation. In a specific embodiment, the oral preparation comprises tablets, sustained-release preparations, capsules, controlled-release preparations, dripping pills and liquid preparations, and the injection administration dosage form comprises intramuscular injection, intravenous injection and intravenous drip.

The dosage form for administration may be a liquid dosage form, a solid dosage form, or a semi-solid dosage form. The liquid dosage forms can be solution (including true solution and colloidal solution), emulsion (including o/w type, w/o type and multiple emulsion), suspension, injection (including water injection, powder injection and infusion), eye drop, nose drop, lotion, liniment, etc.; the solid dosage form can be tablet (including common tablet, enteric coated tablet, buccal tablet, dispersible tablet, chewable tablet, effervescent tablet, orally disintegrating tablet), capsule (including hard capsule, soft capsule, and enteric coated capsule), granule, powder, pellet, dripping pill, suppository, pellicle, patch, aerosol (powder), spray, etc.; semisolid dosage forms can be ointments, gels, pastes, and the like.

The compound can be prepared into common preparations, sustained release preparations, controlled release preparations, targeting preparations and various particle drug delivery systems.

For tableting the compounds of the invention, a wide variety of excipients known in the art may be used, including diluents, binders, wetting agents, disintegrants, lubricants, glidants. The diluent can be starch, dextrin, sucrose, glucose, lactose, mannitol, sorbitol, xylitol, microcrystalline cellulose, calcium sulfate, calcium hydrogen phosphate, calcium carbonate, etc.; the humectant can be water, ethanol, isopropanol, etc.; the binder can be starch slurry, dextrin, syrup, Mel, glucose solution, microcrystalline cellulose, acacia slurry, gelatin slurry, sodium carboxymethylcellulose, methylcellulose, carboxypropylmethylcellulose, ethylcellulose, acrylic resin, carbomer, polyvinylpyrrolidone, polyethylene glycol, etc.; the disintegrant may be dry starch, microcrystalline cellulose, low-substituted hydroxypropyl cellulose, crosslinked polyvinylpyrrolidone, crosslinked sodium carboxymethylcellulose, sodium carboxymethyl starch, sodium bicarbonate and citric acid, polyoxyethylene sorbitol fatty acid ester, sodium dodecyl sulfate, etc.; the lubricant and glidant may be talc, silicon dioxide, stearate, tartaric acid, liquid paraffin, polyethylene glycol, and the like.

The tablets may be further formulated into coated tablets, such as sugar-coated tablets, film-coated tablets, enteric-coated tablets, or double-layer and multi-layer tablets.

To encapsulate the administration units, the active ingredient of the compounds of the invention can be mixed with diluents and glidants and the mixture can be placed directly into hard or soft capsules. Or the effective component of the compound of the invention can be prepared into granules or pellets with diluent, adhesive and disintegrating agent, and then placed into hard capsules or soft capsules. The various diluents, binders, wetting agents, disintegrants, glidants used to prepare the compound tablets of the present invention may also be used to prepare capsules of the compound of the present invention.

In order to prepare the compound of the invention into injection, water, ethanol, isopropanol, propylene glycol or a mixture thereof can be used as a solvent, and a proper amount of solubilizer, cosolvent, pH regulator and osmotic pressure regulator which are commonly used in the field can be added. The solubilizer or cosolvent can be poloxamer, lecithin, hydroxypropyl-beta-cyclodextrin, etc.; the pH regulator can be phosphate, gallate, hydrochloric acid, sodium hydroxide, etc.; the osmotic pressure regulator can be sodium chloride, mannitol, glucose, phosphate, acetate, etc. For example, mannitol and glucose can be added as proppant for preparing lyophilized powder for injection.

In addition, colorants, preservatives, flavors, or other additives may also be added to the pharmaceutical preparation, if desired. For the purpose of administration and enhancing the therapeutic effect, the drug or pharmaceutical composition of the present invention can be administered by any known administration method. The dosage of the pharmaceutical composition of the compound of the present invention to be administered may vary widely depending on the nature and severity of the disease to be prevented or treated, the individual condition of the patient or animal, the route and dosage form of administration, and the like. Generally, a suitable daily dosage range for a compound of the invention is from 0.001 to 400mg/kg body weight. The above-described dosage may be administered in one dosage unit or divided into several dosage units, depending on the clinical experience of the physician and the dosage regimen including the use of other therapeutic means.

The compounds or compositions of the present invention may be administered alone or in combination with other therapeutic or symptomatic agents. When the compound of the present invention is used in a synergistic manner with other therapeutic agents, the dosage thereof should be adjusted according to the actual circumstances.

The invention has the advantages that:

1. the compound shown in the formula I shows an inhibiting effect on tumor cells related to a Wnt/beta-catenin signal channel, so that a new direction is opened for developing antitumor drugs from traditional Chinese medicine sources;

2. the compound shown in the formula I is a brand-new colorectal cancer and Wnt inhibitor;

3. the compound shown in the formula I has the advantages of high selectivity, abundant sources, mature separation technology and the like, so that the compound can become a promising colorectal cancer targeted therapeutic drug.

The invention will be further illustrated with reference to the following specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. Experimental procedures without specific conditions noted in the following examples, generally followed by conventional conditions, such as Sambrook et al, molecular cloning: the conditions described in the Laboratory Manual (New York: Cold Spring Harbor Laboratory Press,1989), or according to the manufacturer's recommendations. Unless otherwise indicated, percentages and parts are by weight.

The reagents and starting materials used in the present invention are commercially available.

Examples

Experimental example 1 inhibition of tumor cell proliferation involving Wnt/beta-catenin signaling pathway by Compounds represented by formula I

Experimental materials: HCT116 cells, HCC827 cells, H460 cells, PC-3 cells, NCL-N87 cells, U-2OS cells, SK-OV3 cells, HEPG2 cells, MDA-MB-231 cells were purchased from the cell bank of Chinese academy of sciences; h1975 cells, ASPC1 cells, PANC1 cells, CFPAC1 cells, BXPC3 cells, SW480 cells were purchased from ATCC; the compound shown in the formula I is purchased from the national center for Standard substances; 4% paraformaldehyde and crystal violet were purchased from Biyunyan.

The experimental steps are as follows: cells were seeded in 6-well plates (1500/well) and compounds of formula I (0, 5, 10, 20, 40. mu.M) were administered after 24h incubation. After 14 days of treatment, the cells were washed 3 times with PBS, fixed for 10min with 4% paraformaldehyde, discarded, washed 3 times with PBS, stained with crystal violet for 10min, washed 3 times with PBS, and observed for cloning.

As a result: the compound shown in formula I has inhibitory effect on 2 kinds of colorectal cancer cells, 3 kinds of non-small cell lung cancer cells, 4 kinds of pancreatic cancer cells, prostate cancer cells, gastric cancer cells, ovarian cancer cells, breast cancer cells, liver cancer cells, osteosarcoma cells and clone formation (shown in figure 1).

EXAMPLE 2 Effect of the Compound of formula I on subcutaneous transplantation of Large intestinal cancer cells (HCT116)

Experimental materials: BALB/c nude mice (four week old), male, purchased from Shanghai Sphere-BiKai laboratory animals, Inc. BALB/c nude mice were bred under SPF-grade conditions.

Solution preparation: and (3) dissolving the compound shown in the formula I by using water as a solvent to prepare suspension.

Cell culture: the state of HCT116 cells was observed daily, and when the cell state was good, i.e., in the logarithmic growth phase, the cells were collected by digestion, 1mL of PBS was added, the cell pellet was carefully washed twice, the cell pellet was resuspended in PBS, and the cell density was adjusted to 5X 10⁶one/mL.

Subcutaneous injection of nude mice: taking the nude mouse to be inoculated, wiping the injection part (right scapula) of the nude mouse with alcohol cotton, blowing the cell suspension again to prevent the cell aggregation, injecting the HCT116 cell suspension into the nude mouse subcutaneously by a 1mL medical injector, each injecting 1X 10⁶0.2mL, and special attention is needed to resuspend the cells before each injection, so as to avoid deviation of the tumorigenic condition caused by uneven cells, and the cells are slowly injected into the subcutaneous space of the right shoulder blade, and the tumorigenic condition is frequently observed and recorded. Tumor volume 1/2 × length × width²。

Grouping experiments: on day 7, the subcutaneous graft tumors grew to reach a palpable state, and were randomly divided into a normal control group and a compound group of formula I (20 mg. kg) according to tumor volume^-1) 10 per group.

Gavage daily of a Compound of formula I (20mg kg) except for the control group^-1) In the normal control group, the mice were sacrificed after administering the normal control group with an equal amount of water daily for 14 consecutive days, and the tumor volume was measured and the tumor weight was weighed. Weigh 3 times per week during dosing.

As a result: after the compound shown in the formula I (20mg/kg) is orally taken by a model animal, the tumor volume and the tumor weight can be reduced to a certain degree, and as shown in figure 2, the compound shown in the formula I can play a role in resisting colorectal cancer when orally taken.

Statistical analysis: the experimental data are expressed in mean ± sd, and the comparison between two groups of samples is performed by t test, and P <0.05 is statistically significant.

EXAMPLE 3 Wnt/beta-catenin pathway inhibition of Compounds of formula I

Experimental materials: 293 cells were purchased from the cell bank of the Chinese academy of sciences. The AB-line zebra fish is purchased from the national zebra fish resource center.

TOP/Flash experiment: 293FT cells were inoculated into 96-well white plates (20000/well), and 24h after plating, TOP/Flash plasmid and Renilla plasmid were administered, 6h after Lipuidambactone (0, 5, 10, 20. mu.M). After 24h of treatment, luciferase activity was assayed.

Zebra fish embryo 'eye loss' experimental procedure: when the AB line zebrafish embryo is 6hpf, the corresponding compounds are administered in groups according to different administration groups. Wherein the model group was administered 6BIO (1. mu.M), and the compound group represented by formula I was administered 6BIO (1. mu.M) simultaneously with the compound group represented by formula I (10. mu.M); the embryo status was then observed at 30hpf and 72 hpf.

The results show that: the compound shown in the formula I can inhibit the transcriptional activity mediated by beta-catenin in cells (figure 3A); the 'eye deletion phenomenon' can occur when the Wnt/beta-catenin pathway is abnormally activated at the initial embryonic development stage of zebra fish, and when the 'eye deletion phenomenon' is rescued after the compound shown in the formula I is given, the compound can be considered to have the Wnt/beta-catenin signal pathway inhibition effect.

The results of this example suggest that the compound represented by formula I has an inhibitory effect on Wnt signaling pathway and can be used as a novel colorectal cancer molecular targeting drug.

By combining the embodiments, the invention adopts a naked mouse model of colorectal cancer HCT116 cell subcutaneous transplanted tumor, and inspects liquidambar formosana hance anti-colorectal cancer drugs, particularly the effect of reducing the volume and the tumor weight of the subcutaneous transplanted tumor, and the result shows that the liquidambar formosana hance oral administration of liquidambar formosana hance can obviously reduce the volume and the tumor weight of the subcutaneous transplanted tumor of mice, and the weight of mice of each administration group has no obvious difference, so a safe, effective and economic solution is provided for the prevention and treatment of colorectal cancer.

All documents referred to herein are incorporated by reference into this application as if each were individually incorporated by reference. Furthermore, it should be understood that various changes and modifications of the present invention can be made by those skilled in the art after reading the above teachings of the present invention, and these equivalents also fall within the scope of the present invention as defined by the appended claims.