CN115969975A - Application of PHGDH inhibitors in prevention and/or treatment of colorectal cancer metastasis - Google Patents

Abstract

The invention belongs to the field of biological medicine, and discloses application of PHGDH serving as a target in inhibiting colorectal cancer metastasis. The invention also discloses an application of the PHGDH inhibitor, the PHGDH inhibitor is used as an active ingredient for inhibiting colorectal cancer metastasis, can prevent and/or treat colorectal cancer metastasis, effectively prolongs the life of a patient while improving the life quality of the patient, and has good clinical application prospect.

Description

Application of PHGDH inhibitors in prevention and/or treatment of colorectal cancer metastasis

technical field

The invention belongs to the field of biomedicine, and relates to the application of a PHGDH inhibitor in inhibiting colorectal cancer metastasis, and is used for preventing and/or treating colorectal cancer metastasis.

Background technique

Colorectal cancer is one of the most common digestive tract tumors, which seriously threatens human health. The liver is the main target organ of colorectal cancer hematogenous metastasis, and about 40% to 70% of colorectal cancer patients will eventually develop liver metastasis and/or colorectal cancer metastases throughout the course of the disease. Some colorectal cancer patients were detected with liver metastases at the time of diagnosis, which were synchronous liver metastases and/or colorectal cancer metastases; in addition, liver metastases were not detected at the time of diagnosis, but in the primary tumor of colorectal cancer Metachronous liver metastases occurred after radical resection. Liver metastasis is the main cause of death in patients with colorectal cancer. Therefore, liver metastasis of colorectal cancer is also an important and difficult point in the treatment of colorectal cancer.

Liver metastasis of colorectal cancer is a complicated process, which involves the invasion of blood vessels by tumor cells from the primary site of colorectum, the invasion of liver by tumor cells (CTCs) in the blood circulation, evading the body's immune killing, and the formation of new metastases in the liver. In order to improve the prospect of treating and preventing colorectal cancer metastasis, it is urgent to develop drugs that can effectively inhibit colorectal cancer metastasis and reduce the high mortality rate caused by colorectal cancer metastasis, which is of great significance.

Contents of the invention

The technical problem to be solved by the present invention is to overcome the deficiencies of the drugs for the treatment of colorectal cancer metastasis in the prior art. Through a large number of experimental screening and research, it is shown that phosphoglycerate dehydrogenase (phosphoglycerate dehydrogenase, PHGDH) inhibitors can be used as inhibitors of colorectal cancer. active ingredient in cancer metastasis. PHGDH, also known as phosphoglycerol dehydrogenase, catalyzes the oxidation of 3-phosphoglycerate to 3-phosphooxypyruvate, which is a key step in the biosynthesis of serine. PHGDH inhibitors can inhibit the enzymatic activity of PHGDH, thereby inhibiting the synthesis of serine and its downstream products. Currently reported PHGDH inhibitors can be used to inhibit the growth of colorectal cancer, but there is no report on the application of PHGDH inhibitors in inhibiting colorectal cancer metastasis. The mechanism of PHGDH inhibitors to inhibit cancer growth is: inhibiting PHGDH prevents the synthesis of serine from glucose, inhibits the synthesis of intracellular nucleotides, and leads to cell cycle arrest. The mechanism by which PHGDH inhibitors inhibit cancer metastasis is: inhibiting PHGDH inhibits the serine synthesis pathway, reduces the content of downstream S-adenosylmethionine (SAM), inhibits the expression of cell migration-related genes, and thus inhibits cancer metastasis.

In order to achieve the above objectives, the present invention aims to provide the use of PHGDH as a target in the preparation of drugs for preventing and/or treating colorectal cancer metastasis.

One aspect of the present invention is to provide the application of PHGDH inhibitors in the preparation of drugs for inhibiting colorectal cancer metastasis, and provide a new method and pharmaceutical composition for preventing and/or treating colorectal cancer metastasis.

One aspect of the present invention is to provide a PHGDH inhibitor or its pharmaceutically acceptable salt, ester, isomer, prodrug, polymorph or solvate and its application in the preparation of a drug for inhibiting colorectal cancer metastasis.

Further preferably, the PHGDH inhibitor is a compound with the molecular formula C ₂₀ H ₂₃ F ₃ N ₄ S (CAS No. 1916571-90-8), the structure of which is shown in Formula I:

Another aspect of the present invention is to provide a pharmaceutical composition for inhibiting colorectal cancer metastasis, said pharmaceutical composition comprising a therapeutically effective amount of the above-mentioned application described above mixed with at least one pharmaceutically acceptable excipient. A PHGDH inhibitor or a pharmaceutically acceptable salt, ester, isomer, prodrug, polymorph or solvate thereof.

In another preferred example, the dosage form of the pharmaceutical composition is selected from: injections, sterile powders for injections, tablets, capsules, formulations, powders, granules, syrups, solutions, tinctures, aerosols, Powder spray, or suppository.

In the present invention, the pharmaceutical composition can also be administered in combination with at least one immune system regulator and/or at least one agent for inhibiting colorectal cancer metastasis.

Another aspect of the present invention is to provide the application of the pharmaceutical composition in the preparation of a medicament for preventing and/or treating colorectal cancer metastasis.

Another aspect of the present invention is to provide an in vitro (preferably non-therapeutic) method of inhibiting colorectal cancer metastasis, said method comprising: administering an effective amount of the above-mentioned PHGDH inhibitor to a subject in need of inhibiting colorectal cancer metastasis or a pharmaceutically acceptable salt, ester, isomer, prodrug, polymorph or solvate thereof.

Compared with the prior art, the beneficial effects of the present invention are:

The present invention creatively uses a PHGDH inhibitor, especially a PHGDH inhibitor represented by formula I, to inhibit colorectal cancer metastasis. Through the in vitro inhibition of colorectal cancer metastasis test, it is found that the PHGDH inhibitor of the present invention has an unexpected effect of inhibiting colorectal cancer metastasis, achieves the purpose of effectively treating colorectal cancer metastasis and improving the prognosis of patients, and complements the treatment of surgery, radiotherapy and chemotherapy. Insufficient means, no toxic side effects. The PHGDH inhibitor has great clinical application value in the preparation of drugs for preventing and/or treating colorectal cancer metastasis, and can be used as an effective method for intervening colorectal cancer metastasis.

Description of drawings

Figure 1 shows the effect of PHGDH inhibitors on the metastasis of colorectal cancer cells in vitro.

Figure 2 is the effect of PHGDH inhibitors on the growth and metastasis of colorectal cancer in mice in vivo. Among them, A is the weight of the subcutaneous tumor; B is the number of metastatic nodules on the liver; C is the metastatic nodules on the liver proved to be tumor tissue by paraffin embedding and HE staining.

Detailed ways

The invention proposes a new active ingredient for the treatment of colorectal cancer metastasis. Combined with the inventor's research on the catalytic effect of PHGDH for many years, through a series of in vivo and in vitro functional experiments, it was surprisingly found that the inhibition of PHGDH can effectively inhibit the metastasis of colorectal cancer cells, and has an excellent effect of inhibiting the metastasis of colorectal cancer. The inventors measured the effect of PHGDH inhibitors on the metastasis of colorectal cancer cells by inoculating colorectal cancer cells in Transwell chambers and staining them in vitro, by constructing a mouse colorectal cancer tumor model, and by measuring the subcutaneous tumor The weight and the number of metastatic nodules on the liver were counted to confirm that PHGDH inhibitors can inhibit the liver metastasis of colorectal cancer.

The invention provides the use of PHGDH as a target in the preparation of drugs for preventing and/or treating colorectal cancer metastasis.

Using PHGDH as a target specifically means: using PHGDH as an action object to screen drugs or preparations to find a drug that can inhibit PHGDH as a candidate drug for the prevention and/or treatment of colorectal cancer metastasis. The PHGDH inhibitor represented by formula I according to the present invention is obtained by screening PHGDH as an action object, and can be used as a drug capable of inhibiting colorectal cancer metastasis. In addition, such as antibody drugs, nucleic acid drugs, etc. can also use PHGDH as the target.

The present invention provides the application of PHGDH inhibitor or its pharmaceutically acceptable salt, ester, isomer, prodrug, polymorph or solvate in the prevention and/or treatment of colorectal cancer metastasis.

Further, the inhibitor is a small molecule compound, an antibody, a nucleic acid or a sequence encapsulated by a lentivirus.

The nucleic acid includes but is not limited to: antisense oligonucleotide, double-stranded RNA (dsRNA), ribozyme, small interfering RNA prepared by endoribonuclease III or short hairpin RNA (shRNA).

The dose of the drug for colorectal cancer metastasis is sufficient to reduce the transcription or translation of human PHGDH gene, or the dose to reduce the expression, activity or function of human PHGDH protein, so that the expression of human PHGDH gene is reduced by at least 50%, Specifically, it may be reduced by 55%, 60%, 65%, 70%, 75%, 80%, 90%, 95%, 99% or 100%.

The medicine must include a PHGDH inhibitor, and use the PHGDH inhibitor as an active ingredient for the aforementioned efficacy.

In the above-mentioned medicine, the active ingredients that exert the aforementioned functions may be only PHGDH inhibitors, and may also contain other molecules that can exert the aforementioned functions.

That is, the PHGDH inhibitor is the only active ingredient or one of the active ingredients of the drug.

The drug can be a single-component substance or a multi-component substance.

The form of the medicament is not particularly limited, and may be in various forms such as solid, liquid, gel, semi-fluid, and aerosol.

The main targets of the drug are mammals. The mammal is preferably a rodent, an artiodactyla, a perissodactyla, a lagomorpha, a primate or the like. The primate is preferably a monkey, ape or human.

Further preferably, the PHGDH inhibitor is a compound with the molecular formula C ₂₀ H ₂₃ F ₃ N ₄ S, the CAS number is 1916571-90-8, and the structure is shown in Formula I:

In the compound of the present invention and its application, the active compound is PHGDH inhibitor or pharmaceutically acceptable salt, ester, isomer, prodrug, polymorph or solvate has the activity of inhibiting colorectal cancer metastasis, and has Metastasis of rectal cancer has prophylactic and/or therapeutic effects.

In the compounds of the present invention and their applications, the salts and esters of the PHGDH inhibitors can be used in the form of pharmaceutically or physiologically acceptable salts or esters. References to "pharmaceutically acceptable salts" generally refer to any salt (usually from say, which means that it is non-toxic, especially as a result of the counterion being non-toxic). These physiologically acceptable salts may be formed with cations or bases, and in the context of the present invention, especially when administered in humans and/or mammals, they are to be understood as consisting of at least A compound, usually an acid (deprotonated), such as a salt of an anion and at least one physiologically tolerated cation, preferably an inorganic cation. In the context of the present invention, salts with alkali metals and alkaline earth metals, and salts with ammonium cations (NH4+), including but not limited to (mono) or (di) sodium, ( Mono) or (di) salts of potassium, magnesium or calcium. These physiologically acceptable salts may also be formed with anions or acids, and in the context of the present invention, especially when administered in humans and/or mammals, they are to be understood as provided by the present invention. At least one compound, usually protonated, such as a salt of a cation and at least one physiologically tolerable anion. The salts and esters of the PHGDH inhibitor include but are not limited to salts or esters formed with the following acids: hydrochloric acid, hydrobromic acid, sulfuric acid, citric acid, tartaric acid, phosphoric acid, lactic acid, pyruvic acid, acetic acid, succinic acid, oxalic acid , fumaric acid, maleic acid, oxaloacetic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, or isethionic acid. Halide salts are also suitable. Other salts include: salts with alkali metals or alkaline earth metals such as sodium, potassium, calcium or magnesium.

In the compound of the present invention and its application, the "prodrug" means that after being taken in an appropriate way, the "prodrug" undergoes metabolism or chemical reaction in the human body to convert into the active PHGDH inhibitor or A pharmaceutically acceptable salt, ester, isomer, prodrug, polymorph or solvate.

In the compounds of the present invention and their applications, the active compounds can also be used in combination with other treatment means, such as surgery, radiotherapy, chemotherapy, and targeted therapy.

In the compounds of the present invention and their applications, when the active compound is used in combination with other therapeutic agents, the active compound is co-administered with other therapeutic agents. "Co-administration" means simultaneous administration via the same or different routes in the same formulation or in two different formulations, or sequential administration via the same or different routes. "Sequential" administration means having a time difference in seconds, minutes, hours or days between the administration of two or more different compounds.

The present invention also provides a pharmaceutical composition, comprising the above-mentioned PHGDH inhibitor or its pharmaceutically acceptable salt, ester, isomer, prodrug, polymorph or solvate, and optionally a One or more pharmaceutically acceptable carriers and media. The acceptable carrier, medium such as sterile water or physiological saline, stabilizer, excipient, antioxidant (ascorbic acid, etc.), buffer (phosphoric acid, citric acid, other organic acids, etc.), preservative, surface Active agents (PEG, Tween, etc.), chelating agents (EDTA, etc.), binders, etc. Moreover, it may also contain other low molecular weight polypeptides; proteins such as serum albumin, gelatin or immunoglobulin; amino acids such as glycine, glutamine, asparagine, arginine and lysine; carbohydrates such as polysaccharides and monosaccharides or Carbohydrates; sugar alcohols such as mannitol or sorbitol. When preparing an aqueous solution for injection, such as physiological saline, isotonic solution containing glucose or other auxiliary drugs, such as D-sorbitol, D-mannose, D-mannitol, sodium chloride, appropriate Solubilizers such as alcohols (ethanol, etc.), polyols (propylene glycol, PEG, etc.), nonionic surfactants (Tween 80, HCO-50) and the like.

In the pharmaceutical composition of the present invention, the PHGDH inhibitor or its pharmaceutically acceptable salt, ester, isomer, prodrug, polymorph or solvate can be a single active ingredient, or can be combined with other active ingredients Combine to form a joint preparation. Preferably, the pharmaceutical composition can also be used in combination with compounds and agents useful for inhibiting colorectal cancer metastasis.

In the pharmaceutical composition of the present invention, the content of the active component (PHGDH inhibitor or its pharmaceutically acceptable salt, ester, isomer, prodrug, polymorph or solvate) is usually a safe and effective amount, the The safe and effective amount should be adjustable for those skilled in the art. For example, the administration amount of the active ingredient usually depends on the body weight of the patient, the type of application, the condition and severity of the disease, for example, as the active ingredient The dosage can usually be 1～1000mg/kg/day, 20～200mg/kg/day, 1～3mg/kg/day, 3～5mg/kg/day, 5～10mg/kg/day, 10～25mg/kg /day, 25～30mg/kg/day, 30～40mg/kg/day, 40～60mg/kg/day, 60～80mg/kg/day, 80～100mg/kg/day, 100～150mg/kg/day , 150-200 mg/kg/day, 200-300 mg/kg/day, 300-500 mg/kg/day, or 500-1000 mg/kg/day.

Those skilled in the art can consider the effective amount to be administered according to the severity of the disease and the health condition and age of the subject. An effective amount will generally vary from 0.01 ng/kg body weight to about 100 mg/kg body weight.

The active ingredient provided by the present invention or the pharmaceutical composition containing the active ingredient can be adapted to any form of administration, which can be oral or parenteral administration, for example, through the lung, through the nose, through the rectum and and/or intravenous injection, more specifically intradermal, subcutaneous, intramuscular, intraarticular, intraperitoneal, pulmonary, buccal, sublingual, nasal, transdermal, vaginal, oral or parenteral administration; injection Administration includes intravenous injection, intramuscular injection, subcutaneous injection, etc., transdermal administration, etc.

As used herein, the dosage form of the pharmaceutical composition is selected from the group consisting of injections, sterile powders for injections, tablets, pills, capsules, lozenges, retorts, powders, granules, syrups, solutions, tinctures, aerosols elixirs, powders, or suppositories. Those skilled in the art can select the appropriate preparation form according to the administration method. For example, the preparation form suitable for oral administration can include but not limited to pills, tablets, chewables, capsules, granules, solutions, drops, etc. formulations, syrups, aerosols or powder sprays, etc. For another example, formulations suitable for parenteral administration may include but not limited to solutions, suspensions, reconstituted dry formulations or sprays, etc., for another example , those suitable for rectal administration can usually be suppositories, and for example, those suitable for injection administration can be injections, sterile powders for injection, and the like.

Among them, tablets, lozenges, pills, capsules, etc. may also contain the following components: binders, such as gums, acacia, corn starch or gelatin; excipients, such as dicalcium phosphate; disintegrants, such as corn Starch, potato starch, alginic acid, etc.; lubricants, such as magnesium stearate; sweetening agents, such as sucrose, lactose or saccharin, or flavoring agents, such as peppermint, oil of wintergreen or cherry flavoring, may be added. When the unit dosage form is a capsule, it may contain, in addition to materials of the foregoing, a liquid carrier. Various other materials may be present as coatings or to modify the physical form of the dosage unit. For example, tablets, pills or capsules may be coated with shellac, sugar or both. A syrup or elixir may contain the active compound, sucrose as a sweetening agent methyl and propylparabens as preservatives, a color and flavoring such as cherry or orange flavor. Any material used in the preparation of any unit dosage form should be pharmaceutically pure and substantially nontoxic in the amounts employed. Additionally, the active compounds can be incorporated into sustained release preparations or formulations.

The present invention also provides a method for preventing and/or treating colorectal cancer metastasis, which comprises administering a PHGDH inhibitor to a subject.

The subject may be a mammal or a mammalian colorectal cancer cell. The mammal is preferably a rodent, an artiodactyla, a perissodactyla, a lagomorpha, a primate or the like. The primate is preferably a monkey, ape or human. The colorectal cancer cells may be colorectal cancer cells in vitro.

The subject may be a patient suffering from colorectal cancer metastasis or an individual for whom prevention and/or treatment of colorectal cancer metastasis is desired. Or the subject is an ex vivo cancer cell of a patient with colorectal cancer metastasis or an individual expected to prevent and/or treat colorectal cancer metastasis.

The PHGDH inhibitor can be administered to the subject before, during and after receiving colorectal cancer metastasis treatment.

As used herein, the colorectal cancer (also known as colorectal cancer, colon cancer, rectal cancer or bowel cancer) refers to cancer when cancer occurs in the colon or rectum (part of the large intestine).

As used herein, the colorectal cancer metastasis refers to the colorectal cancer tumor cells from the primary site invaded into lymphatic vessels, blood vessels or other ways to grow there, forming a tumor of the same type as the primary site tumor. The liver metastasis of colorectal cancer means that colorectal cancer tumor cells invade the liver from the primary site through various possible ways, and form a tumor of the same type as the primary site tumor in the liver.

The present invention further provides a method for alleviating, delaying or inhibiting colorectal cancer metastasis, comprising administering the PHGDH inhibitor or its pharmaceutically acceptable salt, ester, isomer, prodrug, polymorph to an individual in need of treatment Forms or solvates.

Unless otherwise defined, all professional and scientific terms used herein have the same meanings as commonly understood by those skilled in the art. In addition, any methods and materials similar or equivalent to those described can be applied to the method of the present invention. The preferred implementation methods and materials described herein are for demonstration purposes only.

Based on the above research results, further exploration and development of new diagnostic and therapeutic methods targeting this gene can provide more options for the diagnosis and treatment of patients with colorectal cancer metastasis.

PHGDH inhibitors

Refers to molecules that have an inhibitory effect on PHGDH. The inhibitory effect on PHGDH includes but not limited to: inhibiting the expression or activity of PHGDH.

Inhibiting the expression of PHGDH can specifically be inhibiting the transcription or translation of the PHGDH gene, specifically, it can refer to: making the PHGDH gene not transcribed, or reducing the transcriptional activity of the PHGDH gene, or making the PHGDH gene not translating, or reducing the PHGDH gene the translational level of the gene.

Inhibiting the activity of PHGDH refers to reducing the activity of PHGDH. Preferably, the PHGDH activity is reduced by at least 10%, preferably by at least 30%, more preferably by at least 50%, more preferably by at least 70%, and most preferably by at least 90%, compared to before inhibition.

Those skilled in the art can use conventional methods to regulate the gene expression of PHGDH, such as gene knockout, homologous recombination, interfering RNA and the like.

The inhibition of PHGDH gene expression can be verified by PCR and Western Blot detection of expression.

Preferably, compared with the wild type, the expression of PHGDH gene is reduced by at least 10%, preferably by at least 30%, more preferably by at least 50%, more preferably by at least 70%, and most preferably by at least 90%. Jiadi PHGDH gene is not expressed at all.

small molecule compound

In the present invention, it refers to a compound composed of several or dozens of atoms and with a molecular mass below 1000.

Preparation of drugs for preventing or treating colorectal cancer metastasis

Nucleic acid molecules that reduce PHGDH gene expression in colorectal cancer cells; and/or, PHGDH gene interference nucleic acid constructs; and/or, PHGDH gene interference lentiviruses can be used as active ingredients to prepare drugs for preventing or treating colorectal cancer metastasis . Usually, in addition to the active ingredients, the medicine will also include one or more pharmaceutically acceptable carriers or adjuvants according to the requirements of different dosage forms.

"Pharmaceutically acceptable" means that the molecular entities and compositions do not produce adverse, allergic or other adverse reactions when properly administered to animals or humans.

The "pharmaceutically acceptable carrier or excipient" should be compatible with the active ingredient, that is, it can be blended with it without greatly reducing the effect of the drug under normal circumstances. Specific examples of some substances that can be used as pharmaceutically acceptable carriers or excipients are sugars, such as lactose, glucose and sucrose; starches, such as corn starch and potato starch; cellulose and its derivatives, such as sodium methylcellulose, Ethyl cellulose and methyl cellulose; Gum tragacanth powder; Malt; Gelatin; Talc; Solid lubricants such as stearic acid and magnesium stearate; Calcium sulfate; Vegetable oils such as peanut oil, cottonseed oil, sesame oil, olive oils, corn oil, and cocoa butter; polyols, such as propylene glycol, glycerin, sorbitol, mannitol, and polyethylene glycol; alginic acid; emulsifiers, such as Tween; wetting agents, such as sodium lauryl sulfate; colorants; flavoring agents; tableting agents, stabilizers; antioxidants; preservatives; pyrogen-free water; isotonic saline solution; and phosphate buffer, among others. These substances are used as needed to aid the stability of the formulation or to help enhance the activity or its bioavailability or to produce an acceptable mouthfeel or odor in the case of oral administration.

The words "comprising", "containing", etc. mentioned in this article should be understood as inclusive rather than exclusive or exhaustive; that is, "including but not limited to".

The "therapeutically effective dose" mentioned herein generally refers to an amount that can achieve the effect of treating the diseases listed above after an appropriate administration period.

References herein to "therapeutic" and "prophylactic" are to be understood in their broadest sense. The term "therapeutic" does not necessarily imply that the mammal is treated until full recovery. Similarly, "preventive" does not necessarily mean that the object will not eventually be transferred. Accordingly, treatment and prevention include alleviation of the symptoms of a particular condition or preventing or reducing the risk of a particular condition developing. The term "prevention" is understood as reducing the severity of the onset of a particular condition. Treatment may also reduce the severity of pre-existing conditions or the frequency of exacerbations.

In the present invention, the subject or individual for therapeutic or preventive treatment is preferably a mammal, such as but not limited to humans, primates, livestock (such as sheep, cows, horses, donkeys, pigs), pets (such as dogs, cats) , laboratory test animals (such as mice, rabbits, rats, guinea pigs, hamsters) or captured wild animals (such as foxes, deer). The subject is preferably a primate. The subject is most preferably a human.

The active ingredient, pharmaceutical composition and method of the present invention can be used to prevent colorectal cancer metastasis, not only for the early stage of colorectal cancer metastasis to prevent the establishment of cancer lesions in the liver, but also as a preventive treatment in colorectal cancer Apply before transfer. Alternatively, the occurrence of colorectal cancer metastasis can be avoided by taking the active ingredient or pharmaceutical composition in advance.

Before further describing the specific embodiments of the present invention, it should be understood that the protection scope of the present invention is not limited to the following specific specific embodiments; it should also be understood that the terms used in the examples of the present invention are to describe specific specific embodiments, It is not intended to limit the protection scope of the present invention; in the description and claims of the present invention, unless the context clearly indicates otherwise, the singular forms "a", "an" and "the" include plural forms.

test subject

All mice were housed in a pathogen-free facility at the Center for Excellence in Molecular and Cell Science, Chinese Academy of Sciences. Animals were randomly assigned to control and experimental groups. All animal experiments were approved by the Institutional Animal Care and Use Committee (IACUC) of the Center for Excellence in Molecular Cell Science, Chinese Academy of Sciences and complied with all relevant codes of ethics.

Effect of PHGDH inhibitor shown in embodiment 1 formula I on colorectal cancer cell metastasis in vitro

HCT116 cells were treated with the PHGDH inhibitor represented by Formula I (NCT-503, 10 μM), and changes in cell metastasis were detected.

HCT116 (1×10 ⁵ ) cells were inoculated in the upper chamber of an 8-μm pore size Transwell chamber, and cultured in serum-free medium. Among them, the experimental group (+) added 10 μM of PHGDH represented by formula I, and the blank control group (-) Add the corresponding DMSO. Complete medium containing 10% fetal calf serum was used in the culture plate. After 12 hours of culture, the cells in the Transwell chamber were fixed with 4% paraformaldehyde. Remove untransferred cells on the upper part of the filter with a cotton swab, and stain the cells on the lower part of the filter with 0.4% crystal violet. At the same time, cells were plated individually on culture plates without Transwell chambers to determine the total number of attached cells. The relative amount of cells transferred was calculated by dividing the number of transferred cells by the total number of cells, and then further normalized to the control group. For each experiment, the number of cells was counted in 5 random fields under the microscope (magnification 100×), and 3 independent Transwell chambers were analyzed.

HCT116 cells were treated with PHGDH inhibitors, and it was found that PHGDH inhibitors could inhibit the metastasis of HCT116 cells (Figure 1).

1.2 Influence of the PHGDH inhibitor shown in formula I on colorectal cancer liver metastasis in vivo

Fresh tumor tissues from patients with colorectal cancer were obtained from the Shanghai Cancer Center of Fudan University. The surgically excised tumor tissue was immediately placed in DMEM medium supplemented with 2% v/v penicillin/streptomycin, and transferred on ice to a biological safety cabinet for operation. The tumor tissue was cut into 60 mm ³ small pieces, and a suitable tissue was selected and inoculated subcutaneously in the flank of BALB/c nude mice. When the xenografts reached 6 weeks, the mice were operated on and the tumors were dissected, cut into small pieces (60mm ³ ) and reimplanted into a new generation of mice for the construction of colorectal cancer patient-derived xenografts (PDX ) model, one week after the tumor tissue was implanted into the mice, the experimental group (+): intraperitoneally injected the PHGDH inhibitor (NCT-503, 25 mg/kg body weight) into the mice, and the blank control group (-): injected the mice with A reagent of 5% ethanol, 35% polyethylene glycol 300, and 60% 30% hydroxypropyl-β-cyclodextrin in water was intraperitoneally injected into mice. Once a day for 3 consecutive weeks.

Experimental results:

5 weeks after the inoculation, the mice were killed, and the subcutaneous tumors were taken out and weighed. The results are shown in Figure 2A. As can be seen from Figure 2A, the tumor weight of the mice in the experimental group was significantly lower than that of the blank control group, indicating that the PHGDH of the present invention Inhibitors can inhibit the growth of colorectal cancer tissue (p<0.001, with statistical significance). The mouse liver was taken out, and the number of metastatic nodules on the liver was counted, and the results were shown in Figure 2B; the liver tissue was fixed with 4% paraformaldehyde, embedded in paraffin and stained with HE, which proved that the metastases on the liver were indeed Tumor tissue, the results are shown in Figure 2C; it can be seen from Figure 2B and Figure 2C that the PHGDH inhibitor of the present invention can significantly inhibit the liver metastasis of colorectal cancer, with statistically significant differences.

The above description is only a preferred embodiment of the present invention, and is not intended to limit the present invention in any form and in essence. Several improvements and supplements can be made, and these improvements and supplements should also be regarded as the protection scope of the present invention. Those who are familiar with this profession, without departing from the spirit and scope of the present invention, when they can use the technical content disclosed above to make some changes, modifications and equivalent changes of evolution, are all included in the present invention. Equivalent embodiments; at the same time, all changes, modifications and evolutions of any equivalent changes made to the above-mentioned embodiments according to the substantive technology of the present invention still belong to the scope of the technical solution of the present invention.

Claims (10)

1. Use of PHGDH as a target in the preparation of drugs for preventing and/or treating colorectal cancer metastasis. 2. Use of a PHGDH inhibitor or a pharmaceutically acceptable salt, ester, isomer, prodrug, polymorph or solvate thereof in the preparation of a medicament for preventing and/or treating colorectal cancer metastasis. 3. application as claimed in claim 2, is characterized in that, pharmaceutically acceptable salt, ester comprise the salt or ester that described PHGDH inhibitor forms with following acid: hydrochloric acid, hydrobromic acid, sulfuric acid, citric acid , tartaric acid, phosphoric acid, lactic acid, pyruvic acid, acetic acid, succinic acid, oxalic acid, fumaric acid, maleic acid, oxaloacetic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, or isethionic acid. 4. The application according to claim 2, wherein the PHGDH inhibitor can specifically inhibit the expression of PHGDH or inhibit the activity of PHGDH; Wherein, inhibiting the expression of PHGDH refers to inhibiting the transcription or translation of the PHGDH gene, including: making the PHGDH gene not transcribed, or reducing the transcriptional activity of the PHGDH gene, or making the PHGDH gene not translate, or reducing the translation level of the PHGDH gene ; Compared with before inhibition, the expression of PHGDH is decreased by at least 10%; Wherein, inhibiting the activity of PHGDH refers to reducing the activity of PHGDH; the activity of PHGDH is decreased by at least 10% compared with that before the inhibition. 5. The use according to claim 4, characterized in that the expression or activity of PHGDH is reduced by 50% or more. 6. The application according to claim 2, wherein the molecular formula of the PHGDH inhibitor is C ₂₀ H ₂₃ F ₃ N ₄ S, and the structure is shown in Formula I below:

7. The use according to claim 2, characterized in that the metastasis is liver metastasis. 8. The application according to claim 2, wherein the PHGDH inhibitor prevents, inhibits and/or blocks the metastasis of colorectal cancer by inhibiting the activity of PHGDH and inhibiting the serine synthesis pathway. 9. A pharmaceutical composition for preventing and/or treating colorectal cancer metastasis, characterized in that the pharmaceutical composition comprises the PHGDH inhibitor or its Pharmaceutically acceptable salts, esters, isomers, prodrugs, polymorphs or solvates, and pharmaceutically acceptable carriers and media. 10. The pharmaceutical composition according to claim 9, wherein the transfer is liver transfer; and/or, the pharmaceutical composition is injection, sterile powder for injection, tablet, pill, capsule, lozenge elixirs, elixirs, powders, granules, syrups, solutions, tinctures, aerosols, powder sprays, or suppositories.

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