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

Abstract

This invention belongs to the field of biomedicine and discloses the application of PHGDH as a target in inhibiting colorectal cancer metastasis. This invention also discloses the application of PHGDH inhibitors, which, as active ingredients for inhibiting colorectal cancer metastasis, can prevent and / or treat colorectal cancer metastasis, effectively prolonging patient life while improving their quality of life, and have promising clinical application prospects.

Description

Technical Field

[0001] This invention belongs to the field of biomedicine and relates to the application of PHGDH inhibitors in inhibiting colorectal cancer metastasis, for the prevention and / or treatment of colorectal cancer metastasis. Background Technology

[0002] Colorectal cancer is one of the most common digestive tract tumors, seriously threatening human health. The liver is the primary target organ for hematogenous metastasis of colorectal cancer, with approximately 40% to 70% of colorectal cancer patients eventually developing liver metastases and / or colorectal cancer metastases during the course of the disease. Some colorectal cancer patients are diagnosed with liver metastases, known as synchronous liver metastases and / or colorectal cancer metastases; others do not have liver metastases detected at diagnosis but develop metachronous liver metastases after radical resection of the primary colorectal cancer lesion. Liver metastasis is the leading cause of death in colorectal cancer patients; therefore, treating liver metastases in colorectal cancer is both a key focus and a challenge in its treatment.

[0003] Colorectal cancer liver metastasis is a complex process involving tumor cells invading blood vessels from the primary site in the colorectal region, circulating tumor cells (CTCs) evading the body's immune system to invade the liver, and the formation of new metastatic lesions in the liver. To improve the prospects for treating and preventing colorectal cancer metastasis, it is crucial to develop methods that effectively inhibit colorectal cancer metastasis and reduce the high mortality rate caused by it. Summary of the Invention

[0004] The technical problem this invention aims to solve is to overcome the shortcomings of existing drugs for treating colorectal cancer metastasis. Through extensive experimental screening and research, it has been shown that phosphoglycerate dehydrogenase (PHGDH) inhibitors can serve as effective components for inhibiting colorectal cancer metastasis. PHGDH, also known as phosphoglycerate dehydrogenase, catalyzes the oxidation of 3-phosphoglycerate to 3-phosphooxypyruvate, a key step in serine biosynthesis. 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 are no reports on their application in inhibiting colorectal cancer metastasis. The mechanism by which PHGDH inhibitors inhibit cancer growth is: inhibiting PHGDH prevents the synthesis of glucose-derived serine, inhibits intracellular nucleotide synthesis, and thus leads to cell cycle arrest. The mechanism by which PHGDH inhibitors inhibit cancer metastasis is as follows: inhibiting PHGDH suppresses the serine synthesis pathway and reduces the content of downstream S-adenosylmethionine (SAM), thereby inhibiting the expression of cell migration-related genes and thus inhibiting cancer metastasis.

[0005] To achieve the above objectives, the present invention aims to provide the use of PHGDH as a target in the preparation of medicaments for the prevention and / or treatment of colorectal cancer metastasis.

[0006] This invention provides, in one aspect, the use of PHGDH inhibitors in the preparation of drugs for inhibiting colorectal cancer metastasis, and provides novel methods and pharmaceutical compositions for the prevention and / or treatment of colorectal cancer metastasis.

[0007] One aspect of this invention is to provide PHGDH inhibitors or pharmaceutically acceptable salts, esters, isomers, prodrugs, polymorphs or solvates thereof, and their use in the preparation of medicaments for inhibiting colorectal cancer metastasis.

[0008] More preferably, the PHGDH inhibitor has the molecular formula C 20 H 23 The compound of F3N4S (CAS number 1916571-90-8) has the structure shown in Formula I:

[0009]

[0010] Another aspect of the present invention is to provide a pharmaceutical composition for inhibiting colorectal cancer metastasis, the pharmaceutical composition comprising a therapeutically effective amount of a PHGDH inhibitor as described above, or a pharmaceutically acceptable salt, ester, isomer, prodrug, polymorph, or solvate thereof, mixed with at least one pharmaceutically acceptable excipient.

[0011] In another preferred embodiment, the dosage form of the pharmaceutical composition is selected from: injections, sterile powders for injection, tablets, capsules, tinctures, powders, granules, syrups, solutions, tinctures, aerosols, powder inhalers, or suppositories.

[0012] In this invention, the pharmaceutical composition may also be administered in combination with at least one immunomodulator and / or at least one agent that inhibits colorectal cancer metastasis.

[0013] Another aspect of the present invention is to provide the use of the pharmaceutical composition in the preparation of medicaments for the prevention and / or treatment of colorectal cancer metastasis.

[0014] Another aspect of the present invention is to provide a method for inhibiting colorectal cancer metastasis in vitro (preferably non-therapeutic), the method comprising: administering to a subject requiring inhibition of colorectal cancer metastasis an effective amount of the PHGDH inhibitor as described above, or a pharmaceutically acceptable salt, ester, isomer, prodrug, polymorph, or solvate thereof.

[0015] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0016] This invention creatively utilizes PHGDH inhibitors, especially those shown in Formula I, to inhibit colorectal cancer metastasis. In vitro experiments demonstrating the inhibition of colorectal cancer metastasis revealed that the PHGDH inhibitors of this invention produced unexpected effects in inhibiting colorectal cancer metastasis, achieving the goal of effectively treating colorectal cancer metastasis, improving patient prognosis, and compensating for the shortcomings of surgical, radiotherapy, and chemotherapy treatments, without toxic side effects. PHGDH inhibitors have significant clinical application value in the preparation of drugs for the prevention and / or treatment of colorectal cancer metastasis and can serve as an effective method for intervening in colorectal cancer metastasis. Attached Figure Description

[0017] Figure 1 The effect of PHGDH inhibitors on colorectal cancer cell metastasis in vitro.

[0018] Figure 2 This study investigates the effects of PHGDH inhibitors on the growth and metastasis of colorectal cancer in mice in vivo. In the figures, A represents subcutaneous tumor weight; B represents the number of metastatic nodules on the liver; and C represents paraffin-embedded tissue and HE staining confirming that the metastatic nodules on the liver are tumor tissue. Detailed Implementation

[0019] This invention proposes a novel active ingredient for the treatment of colorectal cancer metastasis. Based on the inventors' years of research on the catalytic activity of PHGDH, a series of in vivo and in vitro functional experiments surprisingly revealed that inhibiting PHGDH can effectively inhibit the metastasis of colorectal cancer cells, demonstrating excellent inhibitory effects on colorectal cancer metastasis. In vitro, the inventors determined the effect of the PHGDH inhibitor on the metastasis of colorectal cancer cells by seeding colorectal cancer cells in Transwell chambers and staining them. By constructing a mouse colorectal cancer tumor model, measuring the weight of subcutaneous tumors and counting the number of metastatic nodules in the liver, the inventors confirmed that the PHGDH inhibitor can inhibit liver metastasis of colorectal cancer.

[0020] This invention provides the use of PHGDH as a target in the preparation of medicaments for the prevention and / or treatment of colorectal cancer metastasis.

[0021] The use of PHGDH as a target specifically refers to screening drugs or formulations with PHGDH as the target to identify drugs that can inhibit PHGDH as potential candidates for the prevention and / or treatment of colorectal cancer metastasis. The PHGDH inhibitor shown in Formula I of this invention is obtained through screening with PHGDH as the target and can be used as a drug to inhibit colorectal cancer metastasis. In addition, antibody drugs, nucleic acid drugs, etc., can also target PHGDH.

[0022] This invention provides the use of PHGDH inhibitors or pharmaceutically acceptable salts, esters, isomers, prodrugs, polymorphs or solvates thereof in medicaments for the prevention and / or treatment of colorectal cancer metastasis.

[0023] Furthermore, the inhibitor is a sequence encapsulated by a small molecule compound, antibody, nucleic acid, or lentivirus.

[0024] The nucleic acids include, but are not limited to: antisense oligonucleotides, double-stranded RNA (dsRNA), ribozymes, small interfering RNA or short hairpin RNA (shRNA) prepared by ribonuclease III.

[0025] The dosage of the drug for colorectal cancer metastasis is sufficient to reduce the transcription or translation of the human PHGDH gene, or sufficient to reduce the expression, activity, or function of the human PHGDH protein, so that the expression of the human PHGDH gene is reduced by at least 50%, specifically by 55%, 60%, 65%, 70%, 75%, 80%, 90%, 95%, 99%, or 100%.

[0026] The drug must include a PHGDH inhibitor, and the PHGDH inhibitor is the active ingredient for the aforementioned effects.

[0027] In the aforementioned drug, the active ingredient that performs the aforementioned function may be only a PHGDH inhibitor, or it may include other molecules that can perform the aforementioned function.

[0028] That is, the PHGDH inhibitor is the only active ingredient or one of the active ingredients of the drug.

[0029] The drug may be a single-component substance or a multi-component substance.

[0030] There are no special restrictions on the form of the drug; it can be in various forms such as solid, liquid, gel, semi-liquid, or aerosol.

[0031] The drug is primarily targeted at mammals. Preferred mammals include rodents, even-toed ungulates, perissodactyls, lagomorphs, and primates. Preferred primates include monkeys, apes, or humans.

[0032] More preferably, the PHGDH inhibitor has the molecular formula C 20 H 23 The compound of F3N4S, with CAS number 1916571-90-8, has the structure shown in Formula I:

[0033]

[0034] In the compounds described in this invention and their applications, the active compounds, namely PHGDH inhibitors or pharmaceutically acceptable salts, esters, isomers, prodrugs, polymorphs or solvates, have inhibitory activity against colorectal cancer metastasis and have preventive and / or therapeutic effects on colorectal cancer metastasis.

[0035] In the compounds described in this invention and their applications, the salts or esters of the PHGDH inhibitors may be used in the form of pharmaceutically or physiologically acceptable salts or esters. The term "pharmaceutically acceptable salt" generally refers to any salt that is physiologically tolerable when used appropriately for treatment (particularly when applied or used in humans and / or mammals) (generally meaning that it is non-toxic, particularly as a result of an anti-ion). These physiologically acceptable salts may be formed with cations or bases, and in the context of this invention, particularly when administered in humans and / or mammals, they should be understood as salts formed from at least one compound provided according to this invention, typically an acid (deprotonated), such as an anion, and at least one physiologically tolerable cation (preferably an inorganic cation). Specifically, in the context of this invention, this may include salts formed with alkali metals and alkaline earth metals, and salts formed with ammonium cations (NH4+), specifically including, but not limited to, salts formed with (mono) or (di) sodium, (mono) or (di) potassium, magnesium, or calcium. These physiologically acceptable salts can also be formed with anions or acids, and in the context of this invention, particularly when administered to humans and / or mammals, they should be understood as salts formed from at least one compound provided according to this invention, typically protonated, such as a cation, and at least one physiologically tolerable anion. Salts and esters of the PHGDH inhibitors 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 hydroxyethanesulfonic acid. Salts of halides are also applicable. Other salts include salts formed with alkali metals or alkaline earth metals (such as sodium, potassium, calcium, or magnesium).

[0036] In the compounds and applications described in this invention, the term "prodrug" refers to a substance that, when taken in an appropriate manner, undergoes metabolism or chemical reaction in the human body to transform into the active PHGDH inhibitor or a pharmaceutically acceptable salt, ester, isomer, prodrug, polymorph, or solvate.

[0037] In the compounds and applications described in this invention, the active compounds can also be used in combination with other treatment methods, such as surgery, radiotherapy, chemotherapy, and targeted therapy.

[0038] In the compounds and applications described in this invention, when the active compound is used in combination with other therapeutic agents, the active compound is administered concurrently with the other therapeutic agents. "Concurrent administration" means simultaneous administration in the same formulation or in two different formulations via the same or different routes, or sequential administration via the same or different routes. "Sequential administration" means a time difference, measured in seconds, minutes, hours, or days, between the administration of two or more different compounds.

[0039] The present invention also provides a pharmaceutical composition comprising the PHGDH inhibitor as described above, or a pharmaceutically acceptable salt, ester, isomer, prodrug, polymorph, or solvate thereof, and optionally one or more pharmaceutically acceptable carriers or media. The acceptable carriers or media include, for example, sterile water or physiological saline, stabilizers, excipients, antioxidants (ascorbic acid, etc.), buffers (phosphate, citric acid, other organic acids, etc.), preservatives, surfactants (PEG, Tween, etc.), chelating agents (EDTA, etc.), binders, etc. Furthermore, it may also contain other low molecular weight peptides; proteins such as serum albumin, gelatin, or immunoglobulins; amino acids such as glycine, glutamine, asparagine, arginine, and lysine; sugars or carbohydrates such as polysaccharides and monosaccharides; and sugar alcohols such as mannitol or sorbitol. When preparing aqueous solutions for injection, such as physiological saline, isotonic solutions containing glucose or other adjuvant drugs, such as D-sorbitol, D-mannose, D-mannitol, sodium chloride, appropriate solubilizers such as alcohols (ethanol, etc.), polyols (propylene glycol, PEG, etc.), and nonionic surfactants (Tween 80, HCO-50, etc.) may be used.

[0040] In the pharmaceutical compositions of the present invention, the PHGDH inhibitor or its pharmaceutically acceptable salt, ester, isomer, prodrug, polymorph, or solvate may be a single active ingredient or may be combined with other active ingredients to form a combination formulation. Preferably, the pharmaceutical composition may also be used in combination with compounds and formulations useful for inhibiting colorectal cancer metastasis.

[0041] In the pharmaceutical compositions of this invention, the content of the active ingredient (a PHGDH inhibitor or its pharmaceutically acceptable salt, ester, isomer, prodrug, polymorph, or solvate) is generally a safe and effective amount. This safe and effective amount should be adjustable to those skilled in the art. For example, the dosage of the active ingredient typically depends on the patient's weight, the type of application, the condition and severity of the disease. For instance, the dosage of the active ingredient can typically be 1–1000 mg / kg / day, 20–200 mg / kg / day, 1–3 mg / kg / day, 3–5 mg / kg / day, etc. g / 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, 150~200mg / kg / day, 200~300mg / kg / day, 300~500mg / kg / day, or 500~1000mg / kg / day.

[0042] Those skilled in the art can determine the effective dose based on the severity of the condition and the subject's health status and age. The effective dose typically varies between 0.01 ng / kg body weight and approximately 100 mg / kg body weight.

[0043] The active ingredient or pharmaceutical composition containing the active ingredient provided by this invention can be adapted to any form of administration, including oral or parenteral administration, for example, via pulmonary, nasal, rectal and / or intravenous injection, and more specifically, via intradermal, subcutaneous, intramuscular, intra-articular, intraperitoneal, pulmonary, oral, sublingual, nasal, percutaneous, vaginal, oral or parenteral administration; injection administration includes intravenous injection, intramuscular injection and subcutaneous injection, percutaneous administration, etc.

[0044] As used herein, the dosage form of the pharmaceutical composition is selected from: injections, sterile powders for injection, tablets, pills, capsules, lozenges, tinctures, powders, granules, syrups, solutions, tinctures, aerosols, powder inhalers, or suppositories. Those skilled in the art can select appropriate formulations based on the route of administration. For example, formulations suitable for oral administration may include, but are not limited to, pills, tablets, chewable tablets, capsules, granules, solutions, drops, syrups, aerosols, or powder inhalers; formulations suitable for parenteral administration may include, but are not limited to, solutions, suspensions, rehydrated dry preparations, or sprays; suppositories are typically suitable for rectal administration; and injections and sterile powders for injection are suitable for injection administration.

[0045] Tablets, lozenges, pills, and capsules may also contain the following components: binders, such as gum arabic, corn starch, or gelatin; excipients, such as dicalcium phosphate; disintegrants, such as corn starch, potato starch, or alginic acid; lubricants, such as magnesium stearate; and sweeteners, such as sucrose, lactose, or saccharin, or flavorings, such as peppermint, wintergreen oil, or cherry flavoring. When the unit dosage form is a capsule, it may contain a liquid carrier in addition to the above-mentioned substances. Various other substances may exist in coating form or be used to improve the physical form of the unit dosage form. For example, shellac, sugar, or both may be used to coat tablets, pills, or capsules. Syrups or elixirs may contain active compounds, sucrose as a sweetener, methylparaben and propylparaben as preservatives, colorings, and flavorings, such as cherry or orange flavoring. Any substance used to prepare any unit dosage form should be pharmaceutically pure and substantially non-toxic in the dosage. In addition, active compounds can be incorporated into sustained-release products or formulations.

[0046] The present invention also provides a method for preventing and / or treating colorectal cancer metastasis by administering a PHGDH inhibitor to a subject.

[0047] The target organism can be a mammal or colorectal cancer cells from a mammal. The mammal is preferably a rodent, even-toed ungulate, perissodactyl, lagomorph, or primate. The primate is preferably a monkey, ape, or human. The colorectal cancer cells can be isolated colorectal cancer cells.

[0048] The subject can be a patient with metastatic colorectal cancer or an individual seeking prevention and / or treatment of metastatic colorectal cancer. Alternatively, the subject can be ex vivo cancer cells from a patient with metastatic colorectal cancer or an individual seeking prevention and / or treatment of metastatic colorectal cancer.

[0049] The PHGDH inhibitor can be administered to subjects before, during, and after treatment for colorectal cancer metastasis.

[0050] As used herein, colorectal cancer (also known as colon cancer, rectal cancer, or bowel cancer) refers to cancer that occurs when cancer develops in the colon or rectum (part of the large intestine).

[0051] As used herein, colorectal cancer metastasis refers to colorectal cancer cells invading the lymphatic vessels, blood vessels, or other pathways from the primary site and growing elsewhere, forming a tumor of the same type as the primary tumor. Colorectal cancer liver metastasis refers to colorectal cancer cells invading the liver from the primary site through various possible pathways, forming a tumor of the same type as the primary tumor in the liver.

[0052] The present invention further provides a method for mitigating, delaying, or inhibiting colorectal cancer metastasis, comprising administering the PHGDH inhibitor or a pharmaceutically acceptable salt, ester, isomer, prodrug, polymorph, or solvate thereof to an individual in need of treatment.

[0053] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as are familiar to those skilled in the art. Furthermore, any methods and materials similar to or equivalent to those described herein may be applied to the methods of this invention. The preferred embodiments and materials described herein are for illustrative purposes only.

[0054] Based on the above research results, further exploration and development of new diagnostic and therapeutic methods targeting this gene could provide more options for the diagnosis and treatment of patients with metastatic colorectal cancer.

[0055] PHGDH inhibitors

[0056] This refers to molecules that have an inhibitory effect on PHGDH. Inhibitory effects on PHGDH include, but are not limited to, inhibiting PHGDH expression or activity.

[0057] Inhibiting PHGDH expression can specifically involve inhibiting the transcription or translation of the PHGDH gene. Specifically, this can mean preventing the PHGDH gene from being transcribed, reducing the transcriptional activity of the PHGDH gene, preventing the PHGDH gene from being translated, or reducing the translation level of the PHGDH gene.

[0058] Inhibiting PHGDH activity means reducing PHGDH activity. Preferably, compared to before inhibition, PHGDH activity is reduced by at least 10%, more preferably by at least 30%, even more preferably by at least 50%, more preferably by at least 70%, and most preferably by at least 90%.

[0059] Those skilled in the art can use conventional methods to regulate the gene expression of PHGDH, such as gene knockout, homologous recombination, and interfering RNA.

[0060] The inhibition of PHGDH gene expression can be verified by detecting expression levels using PCR and Western Blot.

[0061] Preferably, compared with the wild type, PHGDH gene expression is reduced by at least 10%, more preferably by at least 30%, even more preferably by at least 50%, even better by at least 70%, and still better by at least 90%, and most preferably by no expression of the PHGDH gene at all.

[0062] Small molecule compounds

[0063] In this invention, it refers to compounds composed of several or dozens of atoms with a molecular mass of less than 1000.

[0064] Preparation of drugs for the prevention or treatment of colorectal cancer metastasis.

[0065] Drugs for the prevention or treatment of colorectal cancer metastasis can be prepared using 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 as active ingredients. Typically, in addition to the active ingredient, the drug may include one or more pharmaceutically acceptable carriers or excipients, depending on the specific dosage form required.

[0066] "Pharmaceutical acceptable" means that when the molecular basis and the composition are properly administered to animals or humans, they do not produce adverse, allergic, or other adverse reactions.

[0067] A "pharmaceutically acceptable carrier or excipient" should be compatible with the active ingredient, meaning it can be miscible with it without significantly reducing the drug's efficacy under normal circumstances. Specific examples of substances that can serve as pharmaceutically acceptable carriers or excipients include sugars such as lactose, glucose, and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium methylcellulose, ethylcellulose, and methylcellulose; tragacanth gum 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 oil, 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 salt solutions; and phosphate buffers, etc. These substances are used as needed to help stabilize the formulation or to improve its activity or bioavailability or to produce an acceptable taste or smell when taken orally.

[0068] The terms "include" and "contain" in this article should be understood as inclusive, without the meaning of exclusivity or exhaustion; that is, "including but not limited to".

[0069] The term "therapeutic effective dose" as used in this article generally refers to a dose that, after an appropriate period of administration, can achieve the therapeutic effect for the diseases listed above.

[0070] The terms "therapeutic" and "preventive" used in this article should be understood in their broadest sense. The term "therapeutic" does not necessarily imply that a mammal receives treatment until it is fully recovered. Similarly, "preventive" does not necessarily mean that the subject will ultimately not transfer. Therefore, treatment and prevention include alleviating the symptoms of a specific condition or preventing or reducing the risk of developing a specific condition. The term "prevention" can be understood as reducing the severity of a specific condition's onset. Treatment can also reduce the severity of an existing condition or the frequency of acute attacks.

[0071] In this invention, the object or individual for therapeutic or preventative treatment is preferably a mammal, such as, but not limited to, humans, primates, livestock (e.g., sheep, cattle, horses, donkeys, pigs), pets (e.g., dogs, cats), laboratory test animals (e.g., mice, rabbits, rats, guinea pigs, hamsters), or captured wild animals (e.g., foxes, deer). The object is preferably a primate. The most preferred object is a human.

[0072] The active ingredients, pharmaceutical compositions, and methods of the present invention can be used to prevent colorectal cancer metastasis. They can be used not only in the early stages of colorectal cancer metastasis to prevent the establishment of cancerous lesions in the liver, but also as a preventative treatment administered before colorectal cancer metastasis. Alternatively, the occurrence of colorectal cancer metastasis can be avoided by pre-administering the active ingredients or pharmaceutical compositions.

[0073] Before further describing specific embodiments of the present invention, it should be understood that the scope of protection of the present invention is not limited to the specific embodiments described below; it should also be understood that the terminology used in the embodiments of the present invention is for describing specific embodiments and not for limiting the scope of protection of the present invention; in the specification and claims of the present invention, unless otherwise expressly stated in the text, the singular forms "a", "an" and "this" include the plural forms.

[0074] Experimental subjects

[0075] All mice were housed in a pathogen-free facility at the Center for Excellence in Molecular 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 ethical guidelines.

[0076] Example 1: Effect of the PHGDH inhibitor shown in Formula I on colorectal cancer cell metastasis in vitro

[0077] HCT116 cells were treated with the PHGDH inhibitor (NCT-503, 10 μM) shown in Formula I, and changes in cell metastasis were detected.

[0078] HCT116 (1×10) 5Cells were seeded in the upper chamber of 8-μm pore size Transwell chambers and cultured in serum-free medium. The experimental group (+) received 10 μM PHGDH (Formula I), while the blank control group (-) received the corresponding DMSO. The culture plates used complete medium containing 10% fetal bovine serum. After 12 hours of culture, cells in the Transwell chambers were fixed with 4% paraformaldehyde. Untransferred cells in the upper part of the filter were removed with cotton swabs, and cells in the lower part of the filter were stained with 0.4% crystal violet. Simultaneously, cells were separately seeded onto culture plates without Transwell chambers to determine the total number of attached cells. The relative transferred cell count was calculated by dividing the number of transferred cells by the total number of cells, and then normalized to the control group. In each experiment, cell counts were performed in five random fields of view under a microscope (100× magnification), and three independent Transwell chambers were analyzed.

[0079] Treatment of HCT116 cells with PHGDH inhibitors revealed that PHGDH inhibitors could inhibit the metastasis of HCT116 cells. Figure 1 ).

[0080] 1.2 Effect of the PHGDH inhibitor shown in Formula I on colorectal cancer liver metastases in vivo

[0081] Fresh colorectal cancer tumor tissue was obtained from the Shanghai Cancer Center of Fudan University. The surgically removed tumor tissue was immediately placed in DMEM medium supplemented with 2% v / v penicillin / streptomycin and transferred on ice to a biosafety cabinet for further processing. The tumor tissue was cut into 60mm pieces. 3 Small pieces of suitable tissue were selected and subcutaneously inoculated into the lateral ventral region of BALB / c nude mice. Six weeks after xenografting, the mice underwent surgery to dissect the tumor, cutting it into small pieces (60 mm). 3 The tumor tissue was then transplanted into a new generation of mice to construct a patient-derived xenograft (PDX) model of colorectal cancer. One week after tumor tissue was implanted into the mice, the experimental group (+) received an intraperitoneal injection of the PHGDH inhibitor (NCT-503, 25 mg / kg body weight), while the blank control group (-) received an intraperitoneal injection of a reagent containing 5% ethanol, 35% polyethylene glycol 300, and 60% hydroxypropyl-β-cyclodextrin aqueous solution. The treatment was administered once daily for 3 consecutive weeks.

[0082] Experimental results:

[0083] Five weeks after inoculation, mice were sacrificed, subcutaneous tumors were removed, and weighed. The results are as follows: Figure 2 As shown in A, by Figure 2As shown in Figure A, the tumor weight in the experimental group mice was significantly lower than that in the blank control group, indicating that the PHGDH inhibitor described in this invention can inhibit the growth of colorectal cancer tissue (p<0.001, which is statistically significant). Mouse livers were removed, and the number of metastatic nodules on the liver was counted and statistically analyzed. The results are as follows: Figure 2 As shown in B; liver tissue was fixed with 4% paraformaldehyde, embedded in paraffin, and stained with hematoxylin and eosin (HE), confirming that the metastases on the liver were indeed tumor tissue, as shown in the results. Figure 2 As shown in C; by Figure 2 B and Figure 2 As can be seen from C, the PHGDH inhibitor described in this invention can significantly inhibit liver metastasis of colorectal cancer, demonstrating statistical significance.

[0084] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any form or substance. It should be noted that those skilled in the art can make various improvements and additions without departing from the method of the present invention, and these improvements and additions should also be considered within the scope of protection of the present invention. Any modifications, alterations, and equivalent changes made by those skilled in the art based on the above-disclosed technical content without departing from the spirit and scope of the present invention are equivalent embodiments of the present invention. Furthermore, any modifications, alterations, and evolutions made to the above embodiments based on the essential technology of the present invention still fall within the scope of the technical solution of the present invention.

Claims

1. The application of PHGDH inhibitors in the preparation of drugs for the prevention and / or treatment of colorectal cancer metastasis; The molecular formula of the PHGDH inhibitor is C 20 H 23 F3N4S, CAS number 1916571-90-8, has the structure shown in Formula I below: Formula I; The metastasis is a liver metastasis.

2. The application as described in claim 1, characterized in that, The PHGDH inhibitor can specifically inhibit the expression of PHGDH or inhibit the activity of PHGDH; Inhibiting PHGDH expression refers to inhibiting the transcription or translation of the PHGDH gene, including: preventing the PHGDH gene from being transcribed, or reducing the transcriptional activity of the PHGDH gene, or preventing the PHGDH gene from being translated, or reducing the translation level of the PHGDH gene; compared to before inhibition, the expression of PHGDH decreases by at least 10%; Inhibiting PHGDH activity means reducing PHGDH activity; compared to before inhibition, PHGDH activity decreases by at least 10%.

3. The application as described in claim 2, characterized in that, The expression or activity of PHGDH was reduced by 50% or more.

4. The application as described in claim 1, characterized in that, The PHGDH inhibitors prevent, inhibit, and / or block the metastasis of colorectal cancer by inhibiting the activity of PHGDH and the serine synthesis pathway.

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