Application of tyrosine in preparation of product for treating hepatocellular carcinoma

Abstract

The invention provides application of tyrosine in preparation of a product for treating hepatocellular carcinoma. The content of tyrosine in human hepatocellular carcinoma cells is reduced or the combination of tyrosine and aminoacyl-tRNA synthetase is blocked, so that the hepatocellular carcinoma can be effectively treated. Another new treatment means different from operation or chemotherapy is provided for patients with hepatocellular carcinoma, and the problem that methods and medicines for effectively treating hepatocellular carcinoma are lacked at present is solved.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of medicine and health, in particular, relates to the application of tyrosine in the preparation of a product for treating hepatocellular carcinoma. BACKGROUND

[0002] Liver cancer is a malignant tumor with high morbidity and mortality worldwide. In 2022, there were about 865,000 new cases of liver cancer worldwide, accounting for 4.3% of all cancer cases, ranking sixth. There were about 758,000 deaths from liver cancer, accounting for 7.8% of all cancer deaths, ranking third, just behind lung cancer and colorectal cancer. Liver cancer includes hepatocellular carcinoma, cholangiocarcinoma, and hepatic angiosarcoma / hepatoblastoma. Among them, hepatocellular carcinoma (HCC) accounts for 75%-85% of the incidence, and the 5-year overall survival rate is less than 20%.

[0003] Current effective treatments for hepatocellular carcinoma include surgical resection, liver transplantation, chemotherapy, and ablation. Surgical resection is the main treatment for early and intermediate hepatocellular carcinoma patients, but most patients have missed the best opportunity for surgical treatment when diagnosed with hepatocellular carcinoma. In addition, hepatocellular carcinoma is not sensitive to radiotherapy and chemotherapy, so targeted drug therapy has become a common treatment for patients with advanced hepatocellular carcinoma. However, the number of available targeted drugs is still very limited.

[0004] Therefore, the present application provides a new treatment for hepatocellular carcinoma. SUMMARY

[0005] The main purpose of the present application is to provide the application of tyrosine in the preparation of a product for treating hepatocellular carcinoma, to solve the problem of lack of effective drugs for treating hepatocellular carcinoma in the prior art.

[0006] In order to achieve the above-mentioned purpose, according to the first aspect of the present application, the application of tyrosine in the preparation of a product for treating hepatocellular carcinoma is provided.

[0007] Further, the above-mentioned method for treating hepatocellular carcinoma is to inhibit the growth of hepatocellular carcinoma cells.

[0008] Further, the above-mentioned product includes a reagent for reducing the content of tyrosine in the hepatocellular carcinoma cells; and / or a reagent for blocking the binding of tyrosine and aminoacyl-tRNA synthetase;

[0009] Preferably, the reagent for reducing the content of tyrosine in hepatocellular carcinoma cells includes a reagent for reducing the uptake of tyrosine by hepatocellular carcinoma cells;

[0010] Preferably, the reagent for blocking the binding of tyrosine and aminoacyl-tRNA synthetase includes a tyrosine analog.

[0011] More preferably, the tyrosine analog is selected from any one or more of tyrosol, resveratrol, or capsaicin.

[0012] Preferably, the agent that reduces the uptake of tyrosine by the hepatocellular carcinoma cells comprises a tyrosine-degrading agent and / or a tyrosine-depleted amino acid composition.

[0013] Preferably, the tyrosine-degrading agent is selected from a tyrosine ammonia-lyase.

[0014] More preferably, the tyrosine ammonia-lyase is derived from Flavobacterium johnsoniae or Rhodotorula glutinis.

[0015] Preferably, the tyrosine-depleted amino acid composition comprises essential amino acids and / or non-essential amino acids.

[0016] wherein the essential amino acids are selected from any one or more of methionine, valine, histidine, isoleucine, leucine, lysine, phenylalanine, threonine, or tryptophan.

[0017] the non-essential amino acids are selected from any one or more of alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, proline, or serine.

[0018] More preferably, the amino acid composition consists of, in parts by weight: 3.61-3.75 methionine, 4.82-5.0 valine, 3.61-3.75 histidine, 4.82-5.00 isoleucine, 7.23-7.50 leucine, 8.43-8.75 lysine, 1.25-4.82 phenylalanine, 4.82-5.00 threonine, 1.20-1.25 tryptophan, 6.02-6.25 alanine, 6.02-6.25 arginine, 3.01-3.13 asparagine, 6.02-6.25 aspartic acid, 2.41-2.50 cysteine, 18.07-18.75 glutamic acid, 3.01-3.13 glutamine, 6.02-6.25 glycine, 3.01-3.13 proline, and 3.01-3.13 serine.

[0019] More preferably, the above amino acid composition consists of, in parts by weight: 3.7-3.75 methionine, 4.94-5.0 valine, 3.70-3.75 histidine, 4.94-5.00 isoleucine, 7.41-7.50 leucine, 8.64-8.75 lysine, 1.25-2.47 phenylalanine, 4.94-5.00 threonine, 1.23-1.25 tryptophan, 6.17-6.25 alanine, 6.17-6.25 arginine, 3.09-3.13 asparagine, 6.17-6.25 aspartic acid, 2.47-2.50 cysteine, 18.52-18.75 glutamic acid, 3.09-3.13 glutamine, 6.17-6.25 glycine, 3.09-3.13 proline, and 3.09-3.13 serine.

[0020] Further, the above use comprises: reducing the content of tyrosine in the cells of the hepatocellular carcinoma and / or blocking the binding of tyrosine to aminoacyl-tRNA synthetase;

[0021] Preferably, the above blocking the binding of tyrosine to aminoacyl-tRNA synthetase comprises: causing the cells of the hepatocellular carcinoma to take in tyrosine analogs;

[0022] More preferably, the above tyrosine analogs are selected from any one or more of: tyrosol, resveratrol, or capsaicin;

[0023] Preferably, the above reducing the content of tyrosine in the cells of the hepatocellular carcinoma comprises reducing the intake of tyrosine by the cells of the hepatocellular carcinoma;

[0024] More preferably, the intake of tyrosine by the cells of the hepatocellular carcinoma is reduced by:

[0025] 1) degrading tyrosine in the components to be taken in by the cells of the hepatocellular carcinoma using a tyrosine-degrading agent; and / or

[0026] 2) causing the cells of the hepatocellular carcinoma to take in an amino acid composition that lacks tyrosine;

[0027] Preferably, the above tyrosine-degrading agent is selected from tyrosine deaminase;

[0028] More preferably, the above tyrosine deaminase is derived from Flavobacterium johnsoniae or Rhodotorula glutinis;

[0029] Preferably, the above amino acid composition that lacks tyrosine comprises essential amino acids and / or non-essential amino acids;

[0030] wherein the essential amino acids are selected from any one or more of: methionine, valine, histidine, isoleucine, leucine, lysine, phenylalanine, threonine or tryptophan;

[0031] the non-essential amino acids are selected from any one or more of: alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, proline or serine;

[0032] Preferably, the amino acid composition consists of, in parts by weight: 3.61-3.75 methionine, 4.82-5.0 valine, 3.61-3.75 histidine, 4.82-5.00 isoleucine, 7.23-7.50 leucine, 8.43-8.75 lysine, 1.25-4.82 phenylalanine, 4.82-5.00 threonine, 1.20-1.25 tryptophan, 6.02-6.25 alanine, 6.02-6.25 arginine, 3.01-3.13 asparagine, 6.02-6.25 aspartic acid, 2.41-2.50 cysteine, 18.07-18.75 glutamic acid, 3.01-3.13 glutamine, 6.02-6.25 glycine, 3.01-3.13 proline and 3.01-3.13 serine;

[0033] More preferably, the amino acid composition consists of, in parts by weight: 3.7-3.75 methionine, 4.94-5.0 valine, 3.70-3.75 histidine, 4.94-5.00 isoleucine, 7.41-7.50 leucine, 8.64-8.75 lysine, 1.25-2.47 phenylalanine, 4.94-5.00 threonine, 1.23-1.25 tryptophan, 6.17-6.25 alanine, 6.17-6.25 arginine, 3.09-3.13 asparagine, 6.17-6.25 aspartic acid, 2.47-2.50 cysteine, 18.52-18.75 glutamic acid, 3.09-3.13 glutamine, 6.17-6.25 glycine, 3.09-3.13 proline and 3.09-3.13 serine.

[0034] To achieve the above object, according to a second aspect of the present application, there is provided a use of an agent for reducing the content of tyrosine in a hepatocellular carcinoma cell in the manufacture of a product for treating hepatocellular carcinoma.

[0035] Further, the agent for reducing the content of tyrosine in a hepatocellular carcinoma cell includes an agent for reducing the intake of tyrosine by the hepatocellular carcinoma cell;

[0036] Preferably, the agent for reducing the uptake of tyrosine by hepatocellular carcinoma cells comprises an agent for degrading tyrosine and / or a tyrosine-deficient amino acid composition.

[0037] Preferably, the agent for degrading tyrosine is selected from the group consisting of tyrosine deaminase;

[0038] Preferably, the tyrosine deaminase is derived from Flavobacterium johnsoniae or Rhodotorula glutinis.

[0039] Preferably, the tyrosine-deficient amino acid composition comprises essential amino acids and / or non-essential amino acids.

[0040] Preferably, the essential amino acids are selected from any one or more of methionine, valine, histidine, isoleucine, leucine, lysine, phenylalanine, threonine or tryptophan.

[0041] Preferably, the non-essential amino acids are selected from any one or more of alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, proline or serine.

[0042] Preferably, the amino acid composition consists of, in parts by weight: 3.61-3.75 methionine, 4.82-5.0 valine, 3.61-3.75 histidine, 4.82-5.00 isoleucine, 7.23-7.50 leucine, 8.43-8.75 lysine, 1.25-4.82 phenylalanine, 4.82-5.00 threonine, 1.20-1.25 tryptophan, 6.02-6.25 alanine, 6.02-6.25 arginine, 3.01-3.13 asparagine, 6.02-6.25 aspartic acid, 2.41-2.50 cysteine, 18.07-18.75 glutamic acid, 3.01-3.13 glutamine, 6.02-6.25 glycine, 3.01-3.13 proline and 3.01-3.13 serine.

[0043] More preferably, the above amino acid composition consists of, in parts by weight: 3.7-3.75 methionine, 4.94-5.0 valine, 3.70-3.75 histidine, 4.94-5.00 isoleucine, 7.41-7.50 leucine, 8.64-8.75 lysine, 1.25-2.47 phenylalanine, 4.94-5.00 threonine, 1.23-1.25 tryptophan, 6.17-6.25 alanine, 6.17-6.25 arginine, 3.09-3.13 asparagine, 6.17-6.25 aspartic acid, 2.47-2.50 cysteine, 18.52-18.75 glutamic acid, 3.09-3.13 glutamine, 6.17-6.25 glycine, 3.09-3.13 proline, and 3.09-3.13 serine.

[0044] To achieve the above object, according to a third aspect of the present application, there is provided a use of an agent that blocks the binding of tyrosine to aminoacyl-tRNA synthetase in the manufacture of a medicament for treating hepatocellular carcinoma.

[0045] Further, the above agent that blocks the binding of tyrosine to aminoacyl-tRNA synthetase comprises a tyrosine analogue;

[0046] Preferably, the above tyrosine analogue is selected from any one or more of tyrosol, resveratrol, or capsaicin.

[0047] To achieve the above object, according to a fourth aspect of the present application, there is provided a medicament for treating hepatocellular carcinoma, the medicament comprising an agent that reduces the content of tyrosine in the cells of the hepatocellular carcinoma and / or an agent that blocks the binding of tyrosine to aminoacyl-tRNA synthetase;

[0048] Preferably, the above agent that reduces the content of tyrosine in the cells of the hepatocellular carcinoma comprises an agent that reduces the uptake of tyrosine by the cells of the hepatocellular carcinoma;

[0049] More preferably, the above agent that reduces the uptake of tyrosine by the cells of the hepatocellular carcinoma comprises an agent that degrades tyrosine and / or a tyrosine-depleted amino acid composition;

[0050] Further preferably, the above agent that degrades tyrosine is selected from tyrosine deaminase;

[0051] Preferably, the above tyrosine deaminase is derived from Flavobacterium johnsoniae or Rhodotorula glutinis;

[0052] Further preferably, the above tyrosine-depleted amino acid composition comprises essential amino acids and / or non-essential amino acids;

[0053] wherein the essential amino acids are selected from any one or more of: methionine, valine, histidine, isoleucine, leucine, lysine, phenylalanine, threonine, or tryptophan;

[0054] the non-essential amino acids are selected from any one or more of: alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, proline, or serine;

[0055] Further preferably, the amino acid composition consists of, in parts by weight: 3.61-3.75 methionine, 4.82-5.0 valine, 3.61-3.75 histidine, 4.82-5.00 isoleucine, 7.23-7.50 leucine, 8.43-8.75 lysine, 1.25-4.82 phenylalanine, 4.82-5.00 threonine, 1.20-1.25 tryptophan, 6.02-6.25 alanine, 6.02-6.25 arginine, 3.01-3.13 asparagine, 6.02-6.25 aspartic acid, 2.41-2.50 cysteine, 18.07-18.75 glutamic acid, 3.01-3.13 glutamine, 6.02-6.25 glycine, 3.01-3.13 proline, and 3.01-3.13 serine;

[0056] More preferably, the amino acid composition consists of, in parts by weight: 3.7-3.75 methionine, 4.94-5.0 valine, 3.70-3.75 histidine, 4.94-5.00 isoleucine, 7.41-7.50 leucine, 8.64-8.75 lysine, 1.25-2.47 phenylalanine, 4.94-5.00 threonine, 1.23-1.25 tryptophan, 6.17-6.25 alanine, 6.17-6.25 arginine, 3.09-3.13 asparagine, 6.17-6.25 aspartic acid, 2.47-2.50 cysteine, 18.52-18.75 glutamic acid, 3.09-3.13 glutamine, 6.17-6.25 glycine, 3.09-3.13 proline, and 3.09-3.13 serine.

[0057] Preferably, the agent that blocks the binding of tyrosine and aminoacyl-tRNA synthetase comprises a tyrosine analog.

[0058] More preferably, the tyrosine analog is selected from any one or more of: tyrosol, resveratrol, or capsaicin.

[0059] To achieve the above object, according to a fifth aspect of the present application, there is provided an amino acid composition consisting of, in parts by weight, 3.61-3.75 methionine, 4.82-5.0 valine, 3.61-3.75 histidine, 4.82-5.00 isoleucine, 7.23-7.50 leucine, 8.43-8.75 lysine, 1.25-4.82 phenylalanine, 4.82-5.00 threonine, 1.20-1.25 tryptophan, 6.02-6.25 alanine, 6.02-6.25 arginine, 3.01-3.13 asparagine, 6.02-6.25 aspartic acid, 2.41-2.50 cysteine, 18.07-18.75 glutamic acid, 3.01-3.13 glutamine, 6.02-6.25 glycine, 3.01-3.13 proline and 3.01-3.13 serine;

[0060] Preferably, the amino acid composition consists of, in parts by weight, 3.7-3.75 methionine, 4.94-5.0 valine, 3.70-3.75 histidine, 4.94-5.00 isoleucine, 7.41-7.50 leucine, 8.64-8.75 lysine, 1.25-2.47 phenylalanine, 4.94-5.00 threonine, 1.23-1.25 tryptophan, 6.17-6.25 alanine, 6.17-6.25 arginine, 3.09-3.13 asparagine, 6.17-6.25 aspartic acid, 2.47-2.50 cysteine, 18.52-18.75 glutamic acid, 3.09-3.13 glutamine, 6.17-6.25 glycine, 3.09-3.13 proline and 3.09-3.13 serine.

[0061] In the present application, the inventors have found that reducing the content of tyrosine in hepatocellular carcinoma cells of human body or blocking the combination of tyrosine and aminoacyl-tRNA synthetase can effectively treat hepatocellular carcinoma. The reagent capable of reducing the content of tyrosine in hepatocellular carcinoma cells of human body or blocking the combination of tyrosine and aminoacyl-tRNA synthetase can be used to prepare a medicament for treating hepatocellular carcinoma. BRIEF DESCRIPTION OF DRAWINGS

[0062] The accompanying drawings, which form a part of the present application, are used to provide further understanding of the present application, and serve as an aid in explaining the exemplary embodiments of the present application and their descriptions, and are not intended as a limitation of the present application. In the drawings:

[0063] Figure 1A A schematic diagram showing the content of amino acids in human normal tissue and hepatocellular carcinoma tissue according to an embodiment of the present application is shown.

[0064] Figure 1BA schematic diagram showing the content of amino acids in normal tissue and hepatocellular carcinoma tissue of mice according to an embodiment of the present application is shown.

[0065] Figure 2A A schematic diagram showing that tyrosine deaminase FjTAL promotes coumaric acid production according to an embodiment of the present application is shown.

[0066] Figure 2B A schematic diagram showing that tyrosine deaminase RgTAL promotes coumaric acid production according to an embodiment of the present application is shown.

[0067] Figure 3A A schematic diagram showing that tyrosine deaminase FjTAL promotes HCC cell death according to an embodiment of the present application is shown.

[0068] Figure 3B A schematic diagram showing that tyrosine deaminase RgTAL treatment promotes HCC cell death according to an embodiment of the present application is shown.

[0069] Figure 4 A schematic diagram showing that tyrosinol treatment promotes HCC cell death according to an embodiment of the present application is shown.

[0070] Figure 5A A schematic diagram showing that SNU398 cells grow rapidly after tyrosine is removed from the culture medium and then supplemented again according to an embodiment of the present application is shown.

[0071] Figure 5B A schematic diagram showing that SNU449 cells grow rapidly after tyrosine is removed from the culture medium and then supplemented again according to an embodiment of the present application is shown.

[0072] Figure 5C A schematic diagram showing that HCCLM3 cells grow rapidly after tyrosine is removed from the culture medium and then supplemented again according to an embodiment of the present application is shown.

[0073] Figure 5D A schematic diagram showing that PLCPRF5 cells grow rapidly after tyrosine is removed from the culture medium and then supplemented again according to an embodiment of the present application is shown.

[0074] Figure 6 A schematic diagram showing the content of tyrosine in mouse liver tissue treated with tyrosine deaminase according to an embodiment of the present application is shown. DETAILED DESCRIPTION

[0075] It should be noted that the embodiments in the present application and the features in the embodiments can be combined with each other without conflict. The present application will be described in detail below with reference to the embodiments.

[0076] Explanation of terms:

[0077] Hepatocellular carcinoma: Full name Hepatocellular carcinoma, commonly known as HCC, is the most common type of chronic liver cancer in adults and is also the most common cause of death in patients with cirrhosis. It occurs in the environment of chronic liver inflammation and is highly related to chronic viral hepatitis infection (hepatitis B or hepatitis C), alcohol or toxin (such as aflatoxin) exposure, etc.

[0078] Hepatocellular carcinoma and hepatoblastoma are two distinct malignant tumors of the liver, with significant differences in pathogenesis, clinical manifestations, treatment strategies, and prognosis.

[0079] HepG2 is a hepatoblastoma, although it was originally used as a model for hepatocellular carcinoma research, it is actually a hepatoblastoma, and it is not appropriate to use it as a model for hepatocellular carcinoma (HCC) (Arzumanian VA, Kiseleva OI, Poverennaya EV. The Curious Case of the HepG2 Cell Line: 40 Years of Expertise. Int J Mol Sci. 2021 Dec 4;22(23):13135. doi: 10.3390 / ijms222313135. PMID: 34884942; PMCID: PMC8658661). Hepatoblastoma cells HepG2 and hepatocellular carcinoma cells have significant differences in cell size, organelle structure, and chromosome number.

[0080] Tyrosine ammonia lyase (TAL): A member of the aromatic amino acid lyase family, often found in plants (including monocotyledonous plants such as sorghum, barley, wheat, oat, rice, corn, sweet cane, parsley, etc.) and microorganisms (basidiomycetes, capsular red bacteria, actinomycetes, Rhodotorula and Trichosporon), which has the function of catalyzing L-tyrosine into coumaric acid and ammonia, and is used for the production of coumaric acid and for the degradation of tyrosine.

[0081] Tyrosinol: A structural analog of tyrosine, which can competitively bind to tyrosyl tRNA synthetase YARS1, to hinder the normal binding of tyrosine, thereby inhibiting tyrosyl-tRNA synthesis and reducing protein synthesis efficiency.

[0082] As mentioned in the background, there is a lack of effective products for treating hepatocellular carcinoma in the prior art. In the present application, the inventors found that the growth of hepatocellular carcinoma cells was significantly inhibited and the survival time of mice with hepatocellular carcinoma was significantly prolonged by depriving tyrosine in cell culture medium and mouse feed, thereby further finding that reducing the content of tyrosine in the human body can effectively treat hepatocellular carcinoma. The mechanism can be used to develop products for treating hepatocellular carcinoma, and thus the protection scheme of the present application is proposed.

[0083] In a first typical embodiment of the present application, the use of tyrosine in the preparation of a product for treating hepatocellular carcinoma is provided. Tyrosine deficiency can effectively inhibit the growth of hepatocellular carcinoma cells, thereby further playing a role in treating hepatocellular carcinoma. Thus, in a preferred embodiment of the present application, the method for treating hepatocellular carcinoma is to inhibit the growth of hepatocellular carcinoma cells.

[0084] Based on this mechanism, a product capable of reducing the content of tyrosine in hepatocellular carcinoma cells or blocking the binding of tyrosine and aminoacyl-tRNA synthetase is developed, which can inhibit the growth of hepatocellular carcinoma cells and thus play a role in treating hepatocellular carcinoma. In a preferred embodiment of the present application, the product comprises a reagent for reducing the content of tyrosine in the hepatocellular carcinoma cells; and / or a reagent for blocking the binding of tyrosine and aminoacyl-tRNA synthetase.

[0085] The reagent for reducing the content of tyrosine in hepatocellular carcinoma cells includes two application effects, one is to reduce the intake of tyrosine by hepatocellular carcinoma cells, and the other is to reduce the content of tyrosine that has been taken in by hepatocellular carcinoma cells. The reagent can reduce the content of tyrosine in hepatocellular carcinoma cells, thereby playing a role in inhibiting the growth of hepatocellular carcinoma cells.

[0086] The method for reducing the content of tyrosine in hepatocellular carcinoma cells is various and can be selected according to different needs. In a preferred embodiment of the present application, the reagent for reducing the content of tyrosine in hepatocellular carcinoma cells comprises a reagent for reducing the intake of tyrosine by hepatocellular carcinoma cells. When the intake of tyrosine by hepatocellular carcinoma cells is reduced, the growth of hepatocellular carcinoma cells is inhibited.

[0087] In a preferred embodiment of the present application, the reagent for reducing the intake of tyrosine by hepatocellular carcinoma cells comprises a reagent for degrading tyrosine and / or an amino acid composition lacking tyrosine. The reagent for degrading tyrosine degrades tyrosine in the component to be taken in by hepatocellular carcinoma cells. The amino acid composition lacking tyrosine directly deprives hepatocellular carcinoma cells of the intake of tyrosine. Both of these reagents can reduce the intake of tyrosine by hepatocellular carcinoma cells, and the growth of hepatocellular carcinoma cells is inhibited when they lack tyrosine.

[0088] It should be noted that the reagent degrading tyrosine and the amino acid composition lacking tyrosine do not distinguish between normal cells and hepatocellular carcinoma cells and other abnormal cells in reducing the content of tyrosine in cells. Although the above two reagents can reduce the content of tyrosine in normal cells, they do not affect the growth of normal cells.

[0089] Tyrosine ammonia lyase has the function of catalyzing L-tyrosine to coumaric acid and ammonia, which can significantly reduce the content of tyrosine. In a more preferred embodiment of the present application, the above-mentioned reagent degrading tyrosine is selected from tyrosine ammonia lyase.

[0090] Any tyrosine ammonia lyase with tyrosine ammonia lyase activity (including wild-type proteins and mutants) from any source is suitable for use in the present application. Further preferably, the above-mentioned tyrosine ammonia lyase is derived from Flavobacterium johnsoniae or Rhodotorula glutinis. These two enzymes are two enzymes with excellent catalytic activity disclosed in the prior art. The application of these two enzymes to the present application can effectively degrade tyrosine in the components to be ingested in hepatocellular carcinoma cells. In addition, the enzyme can also directly enter the hepatocellular carcinoma cells to degrade the tyrosine that has been ingested in the cells.

[0091] In a preferred embodiment of the present application, the amino acid sequence of the above-mentioned tyrosine ammonia lyase derived from Flavobacterium johnsoniae is SEQ ID NO: 3; and the amino acid sequence of the above-mentioned tyrosine ammonia lyase derived from Rhodotorula glutinis is SEQ ID NO: 4. In a more preferred embodiment of the present application, the nucleotide sequence of the coding gene of the above-mentioned tyrosine ammonia lyase derived from Flavobacterium johnsoniae is SEQ ID NO: 1; and the nucleotide sequence of the coding gene of the above-mentioned tyrosine ammonia lyase derived from Rhodotorula glutinis is SEQ ID NO: 2.

[0092] Reducing the exogenous intake of tyrosine by the subject (such as ingesting the amino acid composition lacking tyrosine) can also reduce the content of tyrosine in hepatocellular carcinoma cells.

[0093] In a preferred embodiment of the present application, the tyrosine-deficient amino acid composition comprises essential amino acids and / or non-essential amino acids; wherein the essential amino acids are selected from any one or more of methionine, valine, histidine, isoleucine, leucine, lysine, phenylalanine, threonine or tryptophan; and the non-essential amino acids are selected from any one or more of alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, proline or serine. The tyrosine-deficient amino acid composition can not only meet the nutritional requirements of normal cell growth, but also inhibit the growth of hepatocellular carcinoma cells, thereby further playing a role in treating hepatocellular carcinoma.

[0094] In a preferred embodiment of the present application, the amino acid composition consists of, in parts by weight: 3.61-3.75 methionine, 4.82-5.0 valine, 3.61-3.75 histidine, 4.82-5.00 isoleucine, 7.23-7.50 leucine, 8.43-8.75 lysine, 1.25-4.82 phenylalanine, 4.82-5.00 threonine, 1.20-1.25 tryptophan, 6.02-6.25 alanine, 6.02-6.25 arginine, 3.01-3.13 asparagine, 6.02-6.25 aspartic acid, 2.41-2.50 cysteine, 18.07-18.75 glutamic acid, 3.01-3.13 glutamine, 6.02-6.25 glycine, 3.01-3.13 proline and 3.01-3.13 serine;

[0095] In a preferred embodiment of the present application, the amino acid composition consists of, in parts by weight: 3.7-3.75 methionine, 4.94-5.0 valine, 3.70-3.75 histidine, 4.94-5.00 isoleucine, 7.41-7.50 leucine, 8.64-8.75 lysine, 1.25-2.47 phenylalanine, 4.94-5.00 threonine, 1.23-1.25 tryptophan, 6.17-6.25 alanine, 6.17-6.25 arginine, 3.09-3.13 asparagine, 6.17-6.25 aspartic acid, 2.47-2.50 cysteine, 18.52-18.75 glutamic acid, 3.09-3.13 glutamine, 6.17-6.25 glycine, 3.09-3.13 proline and 3.09-3.13 serine.

[0096] The amino acid composition in the above proportions can effectively inhibit the growth of hepatocellular carcinoma cells.

[0097] Tyrosine analogs function by competing with tyrosine for binding sites to inhibit tyrosine from functioning. In another preferred embodiment of the present application, the agent that blocks the binding of tyrosine to aminoacyl-tRNA synthetase includes a tyrosine analog.

[0098] In a more preferred embodiment of the present application, the tyrosine analog is selected from any one or more of tyrosol, resveratrol, or capsaicin. Among them, tyrosol can compete with tyrosine for binding to tyrosyl tRNA synthetase YARS1, inhibit tyrosine from functioning (including hindering the normal binding of tyrosine, thereby inhibiting tyrosyl-tRNA synthesis and reducing the efficiency of protein synthesis), and affect diseases related to tyrosine metabolism.

[0099] In a preferred embodiment of the present application, the use includes reducing the content of tyrosine in the hepatocellular carcinoma cells and / or blocking the binding of tyrosine to aminoacyl-tRNA synthetase;

[0100] In a preferred embodiment of the present application, the way to reduce the content of tyrosine in the hepatocellular carcinoma cells includes reducing the intake of tyrosine by the hepatocellular carcinoma cells;

[0101] In a more preferred embodiment of the present application, the intake of tyrosine by the hepatocellular carcinoma cells is reduced by: 1) using an agent that degrades tyrosine to degrade tyrosine in the components to be ingested by the hepatocellular carcinoma cells; and / or 2) causing the hepatocellular carcinoma cells to ingest an amino acid composition that lacks tyrosine;

[0102] In a preferred embodiment of the present application, the amino acid composition that lacks tyrosine includes essential amino acids and / or non-essential amino acids; wherein the essential amino acids are selected from any one or more of methionine, valine, histidine, isoleucine, leucine, lysine, phenylalanine, threonine, or tryptophan; and the non-essential amino acids are selected from any one or more of alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, proline, or serine;

[0103] In one preferred embodiment of the present application, the amino acid composition consists of, in parts by weight: 3.61-3.75 methionine, 4.82-5.0 valine, 3.61-3.75 histidine, 4.82-5.00 isoleucine, 7.23-7.50 leucine, 8.43-8.75 lysine, 1.25-4.82 phenylalanine, 4.82-5.00 threonine, 1.20-1.25 tryptophan, 6.02-6.25 alanine, 6.02-6.25 arginine, 3.01-3.13 asparagine, 6.02-6.25 aspartic acid, 2.41-2.50 cysteine, 18.07-18.75 glutamic acid, 3.01-3.13 glutamine, 6.02-6.25 glycine, 3.01-3.13 proline, and 3.01-3.13 serine.

[0104] In one preferred embodiment of the present application, the amino acid composition consists of, in parts by weight: 3.7-3.75 methionine, 4.94-5.0 valine, 3.70-3.75 histidine, 4.94-5.00 isoleucine, 7.41-7.50 leucine, 8.64-8.75 lysine, 1.25-2.47 phenylalanine, 4.94-5.00 threonine, 1.23-1.25 tryptophan, 6.17-6.25 alanine, 6.17-6.25 arginine, 3.09-3.13 asparagine, 6.17-6.25 aspartic acid, 2.47-2.50 cysteine, 18.52-18.75 glutamic acid, 3.09-3.13 glutamine, 6.17-6.25 glycine, 3.09-3.13 proline, and 3.09-3.13 serine.

[0105] In one preferred embodiment of the present application, the agent that degrades tyrosine is selected from tyrosine deaminase. In a more preferred embodiment of the present application, the tyrosine deaminase is derived from Flavobacterium johnsoniae or Rhodotorula glutinis.

[0106] In one preferred embodiment of the present application, the means of blocking the binding of tyrosine and aminoacyl-tRNA synthetase comprises causing the hepatocellular carcinoma cell to take up a tyrosine analog; preferably, the tyrosine analog is selected from any one or more of: tyrosol, resveratrol, or capsaicin.

[0107] In a second typical embodiment of the present application, there is provided a use of an agent for reducing the content of tyrosine in a hepatocarcinoma cell in the manufacture of a product for treating hepatocarcinoma. The agent is capable of inhibiting the growth of hepatocarcinoma cells by reducing the content of tyrosine in the cells, thereby further playing a role in treating hepatocarcinoma. The medicament prepared by using the agent can be used to treat hepatocarcinoma.

[0108] In a preferred embodiment of the present application, the agent for reducing the content of tyrosine in a hepatocarcinoma cell comprises an agent for reducing the intake of tyrosine by the hepatocarcinoma cell;

[0109] In a preferred embodiment of the present application, the agent for reducing the intake of tyrosine by the hepatocarcinoma cell comprises a tyrosine-degrading agent and / or a tyrosine-deficient amino acid composition;

[0110] In a more preferred embodiment of the present application, the tyrosine-deficient amino acid composition comprises essential amino acids and / or non-essential amino acids; wherein the essential amino acids are selected from any one or more of methionine, valine, histidine, isoleucine, leucine, lysine, phenylalanine, threonine or tryptophan; and the non-essential amino acids are selected from any one or more of alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, proline or serine;

[0111] In a preferred embodiment of the present application, the amino acid composition consists of, in parts by weight: 3.61-3.75 methionine, 4.82-5.0 valine, 3.61-3.75 histidine, 4.82-5.00 isoleucine, 7.23-7.50 leucine, 8.43-8.75 lysine, 1.25-4.82 phenylalanine, 4.82-5.00 threonine, 1.20-1.25 tryptophan, 6.02-6.25 alanine, 6.02-6.25 arginine, 3.01-3.13 asparagine, 6.02-6.25 aspartic acid, 2.41-2.50 cysteine, 18.07-18.75 glutamic acid, 3.01-3.13 glutamine, 6.02-6.25 glycine, 3.01-3.13 proline and 3.01-3.13 serine;

[0112] In a preferred embodiment of the present application, the amino acid composition consists of, in parts by weight, 3.7-3.75 methionine, 4.94-5.0 valine, 3.70-3.75 histidine, 4.94-5.00 isoleucine, 7.41-7.50 leucine, 8.64-8.75 lysine, 1.25-2.47 phenylalanine, 4.94-5.00 threonine, 1.23-1.25 tryptophan, 6.17-6.25 alanine, 6.17-6.25 arginine, 3.09-3.13 asparagine, 6.17-6.25 aspartic acid, 2.47-2.50 cysteine, 18.52-18.75 glutamic acid, 3.09-3.13 glutamine, 6.17-6.25 glycine, 3.09-3.13 proline, and 3.09-3.13 serine.

[0113] In a preferred embodiment of the present application, the agent that degrades tyrosine is selected from the group consisting of tyrosine deaminase.

[0114] In a preferred embodiment of the present application, the tyrosine deaminase is derived from Flavobacterium johnsoniae or Rhodotorula glutinis.

[0115] In a third exemplary embodiment of the present application, there is provided a use of an agent that blocks the binding of tyrosine to aminoacyl-tRNA synthetase in the manufacture of a medicament for treating hepatocellular carcinoma.

[0116] In a preferred embodiment of the present application, the agent that blocks the binding of tyrosine to aminoacyl-tRNA synthetase comprises a tyrosine analog. In a more preferred embodiment of the present application, the tyrosine analog is selected from any one or more of tyrosol, resveratrol, or capsaicin.

[0117] In a fourth exemplary embodiment of the present application, there is provided a medicament for treating hepatocellular carcinoma, the medicament comprising an agent that reduces the amount of tyrosine in the cells of the hepatocellular carcinoma and / or an agent that blocks the binding of tyrosine to aminoacyl-tRNA synthetase.

[0118] In a preferred embodiment of the present application, the agent that reduces the amount of tyrosine in the cells of the hepatocellular carcinoma comprises an agent that reduces the uptake of tyrosine by the cells of the hepatocellular carcinoma; in a preferred embodiment of the present application, the agent that reduces the uptake of tyrosine by the cells of the hepatocellular carcinoma comprises an agent that degrades tyrosine and / or an amino acid composition that lacks tyrosine.

[0119] In a more preferred embodiment of the present application, the agent that degrades tyrosine is selected from the group consisting of tyrosine deaminase.

[0120] In a preferred embodiment of the present application, the tyrosine deaminase is derived from Flavobacterium johnsoniae or Rhodotorula glutinis.

[0121] In a more preferred embodiment of the present application, the tyrosine-deficient amino acid composition comprises essential amino acids and / or non-essential amino acids; wherein the essential amino acids are selected from any one or more of methionine, valine, histidine, isoleucine, leucine, lysine, phenylalanine, threonine or tryptophan; and the non-essential amino acids are selected from any one or more of alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, proline or serine.

[0122] In a preferred embodiment of the present application, the amino acid composition consists of, in parts by weight: 3.61-3.75 methionine, 4.82-5.0 valine, 3.61-3.75 histidine, 4.82-5.00 isoleucine, 7.23-7.50 leucine, 8.43-8.75 lysine, 1.25-4.82 phenylalanine, 4.82-5.00 threonine, 1.20-1.25 tryptophan, 6.02-6.25 alanine, 6.02-6.25 arginine, 3.01-3.13 asparagine, 6.02-6.25 aspartic acid, 2.41-2.50 cysteine, 18.07-18.75 glutamic acid, 3.01-3.13 glutamine, 6.02-6.25 glycine, 3.01-3.13 proline and 3.01-3.13 serine.

[0123] In a preferred embodiment of the present application, the amino acid composition consists of, in parts by weight: 3.7-3.75 methionine, 4.94-5.0 valine, 3.70-3.75 histidine, 4.94-5.00 isoleucine, 7.41-7.50 leucine, 8.64-8.75 lysine, 1.25-2.47 phenylalanine, 4.94-5.00 threonine, 1.23-1.25 tryptophan, 6.17-6.25 alanine, 6.17-6.25 arginine, 3.09-3.13 asparagine, 6.17-6.25 aspartic acid, 2.47-2.50 cysteine, 18.52-18.75 glutamic acid, 3.09-3.13 glutamine, 6.17-6.25 glycine, 3.09-3.13 proline and 3.09-3.13 serine.

[0124] In a preferred embodiment of the present application, the agent that blocks the binding of tyrosine to aminoacyl-tRNA synthetase comprises a tyrosine analog. In a more preferred embodiment of the present application, the tyrosine analog is selected from any one or more of tyrosol, resveratrol, or capsaicin.

[0125] In a fifth exemplary embodiment of the present application, an amino acid composition is provided, consisting of, in parts by weight: 3.61-3.75 methionine, 4.82-5.0 valine, 3.61-3.75 histidine, 4.82-5.00 isoleucine, 7.23-7.50 leucine, 8.43-8.75 lysine, 1.25-4.82 phenylalanine, 4.82-5.00 threonine, 1.20-1.25 tryptophan, 6.02-6.25 alanine, 6.02-6.25 arginine, 3.01-3.13 asparagine, 6.02-6.25 aspartic acid, 2.41-2.50 cysteine, 18.07-18.75 glutamic acid, 3.01-3.13 glutamine, 6.02-6.25 glycine, 3.01-3.13 proline, and 3.01-3.13 serine;

[0126] In a preferred embodiment of the present application, the amino acid composition consists of, in parts by weight: 3.7-3.75 methionine, 4.94-5.0 valine, 3.70-3.75 histidine, 4.94-5.00 isoleucine, 7.41-7.50 leucine, 8.64-8.75 lysine, 1.25-2.47 phenylalanine, 4.94-5.00 threonine, 1.23-1.25 tryptophan, 6.17-6.25 alanine, 6.17-6.25 arginine, 3.09-3.13 asparagine, 6.17-6.25 aspartic acid, 2.47-2.50 cysteine, 18.52-18.75 glutamic acid, 3.09-3.13 glutamine, 6.17-6.25 glycine, 3.09-3.13 proline, and 3.09-3.13 serine.

[0127] The present application is further described in connection with the following specific examples which should not be construed as limiting the scope of the application as claimed.

[0128] Example 1 Determination of changes in amino acid levels in hepatocellular carcinoma tissue

[0129] Human hepatocellular carcinoma tissues and normal tissues, mouse hepatocellular carcinoma tissues and normal tissues were detected by gas chromatography-mass spectrometry method, among which human hepatocellular carcinoma and corresponding normal tissue samples were taken from the liver disease department of the fifth medical center of the general hospital of the people's liberation army, and mouse hepatocellular carcinoma tissues were taken from 12-week-old mice with high expression of MYC in hepatocytes. The mouse will spontaneously form hepatocellular carcinoma at 12 weeks, and the normal tissue is taken from the liver of the control mouse in the same cage.

[0130] Specifically: 50 mg of liver tissue was accurately weighed and placed in a tube, 400 μL of methanol-water (v / v=4:1, containing 0.1% formic acid, containing internal standard succinic acid-2,2,3,3-d4) was added to the tube, two small steel balls were added, and then placed in a freezer at-20°C for 2 min, then ground in a grinder (60HZ, 2 min), then ice bath ultrasonic for 10 min, -20°C for 30 min, centrifugation for 10 min (4°C, 12000 rpm), take 300 μL supernatant. Continue to add 300 μL of methanol-water (v / v=4:1, containing 0.1% formic acid, containing internal standard succinic acid-2,2,3,3-d4) to the residue after taking the supernatant, vortex for 30 s, ice bath ultrasonic for 5 min, centrifugation for 10 min (4°C, 12000 rpm), take 300 μL supernatant, combine the supernatant to a total of 600 μL, vortex and take 100 μL supernatant in a wide buffer tube and dry.

[0131] 80 μL of methoxyamine hydrochloride pyridine solution (15 mg / mL) was added to a glass derivative vial, vortexed for 2 min, then subjected to oximation reaction in a shaking incubator at 37°C (60 min), and then 80 μL of BSTFA derivatization reagent and 20 μL of n-hexane were added to the sample. Vortex for 2 min, then react at 70°C for 60 min, take out the sample, and place it at room temperature for 30 min, then perform GC-MS metabolomics analysis.

[0132] The chromatographic method is: DB-5MS capillary column (30 m x 0.25 mm x 0.25 μm, Agilent J&W Scientific, Folsom, CA, USA), carrier gas is high-purity helium (purity not less than 99.999%), flow rate is 1.2 mL / min, temperature of injection port is 260°C. Injection volume is 1 μL, no split injection, solvent delay is 4 min.

[0133] Programmed temperature: initial temperature of the column oven was 50 °C for 0.5 min, programmed to 125 °C at 15 °C / min for 2 min, 8 °C / min to 210 °C for 2 min, 11 °C / min to 270 °C for 1 min, 25 °C / min to 305 °C for 3 min. Mass spectrometry method: electron impact ion source (EI), ion source temperature 300 °C, transfer line temperature 280 °C, scan mode was selected reaction monitoring scan (SRM), mass scan range: m / z: 40-600.

[0134] The experimental results are shown in Figure 1A and Figure 1B It was found that tyrosine was significantly reduced in human and mouse hepatocellular carcinoma tissues compared with normal tissues, while other amino acids such as cysteine did not change significantly. Therefore, we speculate that hepatocellular carcinoma cells may highly utilize tyrosine, resulting in a decrease in the content of tyrosine in hepatocellular carcinoma tissues.

[0135] Example 2 Limiting tyrosine levels inhibits the proliferation of hepatocellular carcinoma cells without affecting the growth of normal liver cells

[0136] 1. Experimental materials:

[0137] The difference between the control group medium (containing tyrosine) and the experimental group medium (not containing tyrosine) is only that tyrosine is missing in the control group medium based on the control group medium. The composition of the control group medium is shown in Table 1. The control group medium was purchased from Procell, model PM150210. The experimental group medium was purchased from Procell.

[0138] Table 1

[0139]

[0140]

[0141] 2. Experimental method

[0142] The proliferation of four hepatocellular carcinoma cell lines (SUN398, SUN449, HCCLM3 and PLCPRF5; all purchased from Shanghai Fuheng Biotechnology Co., Ltd., model numbers FH0068, FH0816, FH0096 and FH0064, respectively) and human normal liver cells THLE-2 cells (purchased from Shanghai Fuheng Biotechnology Co., Ltd., model number FH1249) in the experimental group (without tyrosine) was determined by the MTT method. Specifically, hepatocellular carcinoma cells and normal liver cells were seeded in 96-well cell culture dishes, and after 24 hours, the culture medium was replaced with control group medium and experimental group medium. Then, after 72 hours, 10 μL of MTT (1 mg / mL) was added to each well, and after 6 hours of reaction at 37°C, the supernatant was aspirated, 100 μL of DMSO solution was added, and after 10 minutes of reaction at 37°C, the absorbance OD value was determined by a microplate reader at a wavelength of 570 nm.

[0143]

[0144] 3. Experimental results

[0145] The cell inhibition rate was calculated according to the above formula, and the results are shown in Table 2 below:

[0146] Table 2: Inhibition rates of four hepatocellular carcinoma cells

[0147]

[0148] The experimental results show that after tyrosine deprivation, the growth of the four hepatocellular carcinoma cells is significantly inhibited, with an inhibition rate of more than 82%, and the highest inhibition rate reaches 87.8%, while the normal liver cells are not significantly affected.

[0149] Experimental steps of the backfill experiment: The proliferation of four hepatocellular carcinoma cell lines (SUN398, SUN449, HCCLM3 and PLCPRF5) in the experimental group (without tyrosine group and tyrosine backfill group) was determined by the MTT method.

[0150] Specifically, hepatocellular carcinoma cells were seeded in 96-well cell culture dishes, and were recorded as 0 hours, and were divided into three groups, namely (1) control group, (2) tyrosine deficiency group and (3) tyrosine deficiency for 48 hours backfill group. 10 μL of MTT (1 mg / mL) was added to each group (three parallel wells), and after 6 hours of reaction at 37°C, the supernatant was aspirated, 100 μL of DMSO solution was added, and after 10 minutes of reaction at 37°C, the absorbance OD value was determined by a microplate reader at a wavelength of 570 nm, which was the absorbance of the 0 hour cells.

[0151] Meanwhile, MTT was added every 24 hours to detect the absorbance of the three groups. The third group was supplemented with tyrosine after 48 hours of culture, and the absorbance of the cells was continuously detected. The experimental results are shown in Tables 1-1 and 1-2. Figure 5A 、 Figure 5B 、 Figure 5C and Figure 5D As shown in Tables 1-1 and 1-2, tyrosine deprivation significantly inhibited the proliferation ability of four hepatocellular carcinoma cell lines, and after the supplementation of tyrosine, the proliferation ability of hepatocellular carcinoma cells was quickly recovered, indicating the necessity of tyrosine for the proliferation of hepatocellular carcinoma.

[0152] Example 3 Tyrosine deprivation reduces the occurrence of mouse hepatocellular carcinoma

[0153] 1. Experimental materials

[0154] Mouse model: HCCLM3 hepatocellular carcinoma cells were used to construct a mouse liver orthotopic tumor. The control group of the present application is a normal diet group (containing tyrosine in the feed), and the experimental group is a tyrosine deprivation group (containing no tyrosine in the feed). The difference between the experimental group and the control group is that the experimental group does not contain tyrosine in the feed that the control group contains.

[0155] Among them, the composition of the feed of the control group is shown in Tables 3-1 and 3-2, Table 3-1 represents the composition of amino acids in the feed, and Table 3-2 represents the composition of non-amino acids in the feed. The feed of the control group of the present application is purchased from Jiangsu Cooperation Pharmaceutical Biological Engineering Co., Ltd., and the model number is XTAA. The feed of the experimental group of the present application is purchased from Jiangsu Cooperation Pharmaceutical Biological Engineering Co., Ltd.

[0156] Table 3-1

[0157] Ingredients gm kcal L-Methionine 6 24 L-Alanine 10 40 L-Arginine 10 40 L-Asparagine-H2O (Asparagine-Aqueous) 5 20 L-Aspartate 10 40 10 40 L-Cystine 4 16 L-Glutamic Acid 30 120 L-Glutamine 5 20 Glycine 10 40 L-Histidine-HCl-H2O (histidine hydrochloride-aqueous solution) 6 24 L-Isoleucine 8 32 L-Leucine 12 48 L-Lysine-HCl (Lysine hydrochloride) 14 56 L-Phenylalanine 8 2 L-Proline 5 20 L-Serine 5 20 L-Threonine 8 32 L-Tryptophan (tryptophan) 2 8 L-Tyrosine 4 16 L-Valine 8 32 Total L-Amino Acids 170 680

[0158] Table 3-2

[0159] Element gm (%) kcal (%) protein 17.00％ 17.56％ carbohydrate 68.55％ 70.82％ Fat 5.00％ 11.62％ Total / 100.00％ kcal / gm (kilocalories per gram) 3.872 / Ingredients gm kcal Corn Starch 550.5 2202 Maltodextrin 10 125 500 Cellulose 50 0 corn oil 50 450 Mineral Mix S10001 35 0 Sodium bicarbonate 7.5 0 Mixed Vitamin V10001 10 40 Hydrocholine tartrate 2 0 total 1000 3872

[0160] In the table, gm represents the abbreviation of grams; kcal / gm refers to the energy value of kilocalories (kcal) per gram of food.

[0161] 2. Experimental method

[0162] Using 4-week-old Bal / bc nude mice, the mice were anesthetized, the abdominal skin was cut, the liver was exposed, then HCCLM3 cells were injected into the liver lobule through a syringe, and finally the liver was returned to its original position, the skin was sutured, and the mouse was waited to wake up. After the mouse woke up, the mouse was divided into two groups (control group and experimental group), and the corresponding feed was replaced, and after 4 weeks of feeding, the mouse was sacrificed, and the number of tumors and the maximum diameter of the tumor were counted.

[0163] 3. Experimental results

[0164] The results are shown in Table 4. Restricting tyrosine in the diet of mice significantly reduced the average number of tumors on the liver (81.17 vs. 41.83, p<0.0001, n=6) and the average maximum diameter (14.25 mm vs. 7.51 mm, p=0.001, n=6).

[0165] Table 4. Effect of tyrosine restriction on HCCLM3 cell tumorigenesis in situ in the liver

[0166] Number of tumors Tumor diameter (mm) Control group (feed containing tyrosine) 81.17 14.25 Experimental group (feed without tyrosine) 41.83 7.51

[0167] The experimental results show that in the mouse hepatocellular carcinoma model, the number of tumors is significantly reduced after limiting the intake of tyrosine compared to the control group, and the tumor diameter is reduced by 50%. There is no significant difference in the body weight of mice between the control group and the experimental group, indicating that whether tyrosine is taken in does not affect the normal growth of mice.

[0168] Example 4. Effect of tyrosine deprivation on the survival of mice with hepatocellular carcinoma

[0169] The feed components of the control group (containing tyrosine in the feed) and the experimental group (not containing tyrosine in the feed) are the same as in Example 3. The experimental method is the same as in Example 5. The experimental results are shown in Table 5. Restricting tyrosine in the feed of mice with hepatocellular carcinoma can significantly prolong the average survival time of mice (56.25 days vs. 83.5 days, p=0.0008, n=8).

[0170] Table 5. Effect of tyrosine restriction on the survival of mice with hepatocellular carcinoma

[0171] Mouse survival time (days) Control group (feed containing tyrosine) 56.25 Experimental group (feed without tyrosine) 83.5

[0172] Example 5. Phenylalanine deprivation reduces the occurrence of hepatocellular carcinoma in mice

[0173] Since phenylalanine can be converted into tyrosine in the body, the applicant further considers reducing the content of phenylalanine in the composition to explore the therapeutic potential of phenylalanine deprivation for hepatocellular carcinoma.

[0174] This example designs 1 control group and 5 experimental groups, respectively: control group (Phe 100% diet + Tyr 100% diet); experimental group 1 (Phe 100% diet + Tyr 0% diet); experimental group 2 (Phe 75% diet + Tyr 0% diet); experimental group 3 (Phe 50% diet + Tyr 0% diet); experimental group 4 (Phe 25% diet + Tyr 0% diet); experimental group 5 (Phe 0% diet + Tyr 0% diet). The experimental groups are all based on the control group and only reduce tyrosine and / or phenylalanine.

[0175] wherein "Phe 100% diet + Tyr 100% diet" means that the content of phenylalanine in the feed of this group is 100% of the content of phenylalanine in the feed shown in Table 3-1 and the content of tyrosine is 100% of the content of tyrosine in the feed shown in Table 3-1. "Phe 100% diet + Tyr 0% diet" means that the content of phenylalanine in the feed of this group is 100% of the content of phenylalanine in the feed shown in Table 3-1 and the feed of this group does not contain tyrosine. "Phe 75% diet + Tyr 0% diet" means that the content of phenylalanine in the feed of this group is 75% of the content of phenylalanine in the feed shown in Table 3-1 and the feed of this group does not contain tyrosine. The content of phenylalanine and tyrosine of other experimental groups are analogously. The experimental results are shown in Table 6 below.

[0176] Table 6

[0177] Number of tumors Tumor diameter (mm) Control group (Phe 100% diet + Tyr 100% diet) 89.7 14.94 Experimental group 1 (Phe 100% diet + Tyr 0% diet) 50.3 8.28 Experimental group 2 (Phe 75% diet + Tyr 0% diet) 40.7 6.93 Experimental group 3 (Phe 50% diet + Tyr 0% diet) 21.8 4.30 Experimental group 4 (Phe 25% diet + Tyr 0% diet) 23.8 4.27 Experimental group 5 (Phe 0% diet + Tyr 0% diet) / /

[0178] Note: Experimental group 5 was terminated because the mice in this group showed a significant decrease in body weight, indicating that the lack of both phenylalanine and tyrosine affected the normal survival of the mice.

[0179] The experimental results showed that compared with the control group, experimental group 1 could significantly reduce the average number of tumors (89.7 vs. 50.3, p<0.0001) and the average maximum diameter of tumors (14.94 vs. 8.28, p<0.0001).

[0180] Compared with experimental group 1, the average number of tumors (50.3 vs. 40.7, p=0.0005) and the average maximum diameter of tumors (8.28 vs. 6.93, p=0.028) in experimental group 2 were significantly reduced.

[0181] Compared with experimental group 2, further increasing the proportion of phenylalanine restriction (50% restriction in experimental group 3), the average number of tumors (40.7 vs. 21.8, p=0.0005) and the average maximum diameter of tumors (6.93 vs. 4.30, p=0.0006) were significantly reduced.

[0182] However, compared with experimental group 3, further reducing the intake of phenylalanine (experimental group 4) did not further improve the therapeutic effect, and the average number of tumors (21.8 vs. 23.8, p>0.05) and the average maximum diameter of tumors (4.23 vs. 4.06, p>0.05) were not significantly different. Further reducing the intake of phenylalanine to 0 caused a significant decrease in body weight of the mice, indicating that hepatocellular carcinoma requires the intake of phenylalanine.

[0183] In summary, the preferred prescription of 50% phenylalanine intake combined with 0% tyrosine intake in mice showed a significant effect on the treatment of hepatocellular carcinoma.

[0184] Example 6 Detection of tyrosine degradation activity of tyrosine ammonia lyase (TAL)

[0185] FjTAL from Flavobacterium johnsoniae and RgTAL from Rhodotorula glutinis were extracted for testing of tyrosine degradation ability.

[0186] In order to facilitate expression in E. coli, the coding genes of the two enzymes (GenBank Accession No. WP_012023194, FjTAL) and (GenBank Accession No. KF765779.1, RgTAL) were optimized according to the codon bias of E. coli, and the optimized nucleotide sequences were SEQ ID NO: 1 and SEQ ID NO: 2, respectively. Among them, the amino acid sequence of the protein encoded by the gene FjTAL is SEQ ID NO: 3; the amino acid sequence of the protein encoded by the gene RgTAL is SEQ ID NO: 4.

[0187] The nucleotide sequence was synthesized by Suzhou Jinyuzhi Biotechnology Co., Ltd., and then cloned into the pET22b-(+) vector between the NdeI and BamHI enzyme cutting sites, finally obtaining two recombinant plasmids pET22b-(+)-FjTAL and pET22b-(+)-RgTAL. Then the recombinant plasmid was transformed into E. coli BL21 (DE3) competent cells.

[0188] After overnight culture, a single colony was picked and inoculated into 6 mL of TB medium containing 50 μg / mL of ampicillin, and cultured at 37°C and 220 rpm / min for 12 hours, then transferred to 1 L of TB medium containing 50 μg / mL of ampicillin, and cultured at 37°C and 220 rpm / min for 12 hours. Then IPTG inducer (final concentration 0.4 mmol / L) was added, and cultured at 25°C and 220 rpm / min for 22 hours. Finally, the bacterial bodies were collected at 4°C and 4000 rpm / min, and the expressed protein was extracted.

[0189] The bacterial cells were resuspended in buffer (50 mM potassium phosphate, 300 mM NaCl, 5 mM imidazole, pH 8.0) and then disrupted by a high-pressure homogenizer. After disruption, the homogenate was centrifuged at 13,000 rpm / min at 4°C to collect the supernatant. The supernatant was placed in ice and then treated with an ultrasonic disrupter at 350 W for 10 min. The protein was purified using a His Trap FF purification column, which was activated with a resuspension solution (containing 500 mM imidazole) before purification. After loading the sample onto the purification column, the protein was eluted with Ni-Elution buffer (50 mM imidazole, NaCl and NaH2PO4-2H2O, pH 7.9-8.1). The presence of the target protein in the eluate was then detected by SDS-PAGE. The eluate was further purified by a molecular sieve. Finally, the supernatant was placed in the inner chamber of a concentration tube and centrifuged at 4°C at 3,500 rpm / min for 10 min to obtain concentrated protein solution.

[0190] The concentrations of FjTAL and RgTAL were detected by BCA method, which were 5.43 mg / mL and 6.28 mg / mL, respectively. The molar molecular weights of the two enzymes were 54.668 kda and 76.053 kda, respectively, and thus the corresponding molar concentrations were 99.3 μM and 82.6 μM, respectively. The abilities of the two enzymes to degrade tyrosine were then detected. The degradation product of tyrosine, coumaric acid, had the maximum absorbance at a wavelength of 310 nm, and thus the absorbance of the reaction solution at 310 nm was detected by an enzyme marker. The reaction conditions were as follows: 250 μM of tyrosine was added to the reaction buffer (50 mM Tris-HCl, pH 9.2), and TAL enzyme was also added so that the final concentration of TAL enzyme in the reaction solution was 0 nM, 0.05 μM, 0.1 μM, 0.25 μM, 0.5 μM, 0.75 μM and 1 μM, respectively. Then the reaction was carried out at 37°C for 1 hour, and the absorbance of the solution was detected by an enzyme marker.

[0191] The experimental results are shown in Tables 1 and 2. Figure 2A and Figure 2B As shown in Tables 1 and 2, the production of coumaric acid gradually increased with the increase of the concentrations of FjTAL and RgTAL after 1 hour of reaction, indicating that both enzymes can effectively degrade tyrosine into coumaric acid.

[0192] Example 7 Tyrosine ammonia lyase (TAL) promotes the death of hepatocellular carcinoma cells

[0193] Purpose of the experiment: To determine the median lethal concentration of FjTAL and RgTAL in causing the death of HCC cells.

[0194] Experimental method: The lethal effect of two TAL enzymes on four hepatocellular carcinoma cell lines (SUN398, SUN449, HCCLM3 and PLCPRF5) was determined by MTT method. Specifically, hepatocellular carcinoma cells were seeded in 96-well cell culture dishes, and after 24 hours, fresh culture medium containing TAL enzymes (see Table 1 for formulation) was added. After 48 hours, 10 μL of MTT (1 mg / mL) was added to each well. After 6 hours of reaction at 37°C, the supernatant was removed, and 100 μL of DMSO solution was added. After 10 minutes of reaction at 37°C, the absorbance OD value was determined by a microplate reader at 570 nm. The absorbance of different TAL concentrations was normalized to the control group, and then plotted to calculate the median lethal concentration (IC50).

[0195] Experimental results: The results are shown in Figures 1 and 2, and both FjTAL and RgTAL can effectively kill hepatocellular carcinoma cells, with a median lethal concentration of about 0.5-0.7 μM, showing a low drug treatment concentration. Figure 3A and Figure 3B

[0196] Example 8 Tyrosine ammonia lyase (TAL) inhibits the development of mouse hepatocellular carcinoma

[0197] Purpose of implementation: To determine the effect of FjTAL and RgTAL on the development of mouse hepatocellular carcinoma

[0198] Experimental method: Four-week-old Bal / bc nude mice were used. After the mice were anesthetized, the abdominal skin was cut to expose the liver, and then HCCLM3 cells were injected into the liver lobules using a syringe. Finally, the liver was returned to its original position, the skin was sutured, and the mice were allowed to wake up. After the mice woke up, they were divided into three groups: a control group, an experimental group 1 (FjTAL treatment), and an experimental group 2 (RgTAL treatment). The experimental group mice were injected intraperitoneally with the corresponding enzyme every two days, with an enzyme dosage of 25 mg / kg, while the control group was injected intraperitoneally with normal saline. After 4 weeks of feeding, the mice were sacrificed, and the number of tumors and the maximum diameter of the tumors were counted.

[0199] Experimental results: The results are shown in Table 7. FjTAL and RgTAL treatment can effectively inhibit the number (76.54 vs. 54.87 and 38.12, p<0.0001) and size (13.88 vs. 8.04 and 7.56, p<0.0001) of liver tumors in mice.

[0200] Table 7 Effect of tyrosine ammonia lyase treatment on HCCLM3 cell tumor formation in situ in the liver

[0201] Number of tumors Tumor diameter (mm) Control group (saline group) 76.54 13.88 Experimental Group 1 (FjTAL Group) 45.87 8.04 Experimental group 2 (RgTAL group) 38.12 7.56

[0202] ​Example 9 Tyrosinol treatment promotes hepatocellular carcinoma cell death

[0203] Objective: To determine the median lethal concentration of Tyrosinol that causes HCC cell death.

[0204] Method: The lethal effect of Tyrosinol treatment on 4 hepatocellular carcinoma cell lines (SUN398, SUN449, HCCLM3 and PLCPRF5) was determined by MTT method. Specifically, hepatocellular carcinoma cells were seeded in 96-well cell culture dishes, and after 24 hours, fresh medium containing Tyrosinol was added. Then, after 48 hours, 10 μL of MTT (1 mg / mL) was added to each well. After 6 hours of incubation at 37°C, the supernatant was removed, and 100 μL of DMSO solution was added. After 10 minutes of incubation at 37°C, the absorbance OD value was determined using a microplate reader at a wavelength of 570 nm. The absorbance values obtained under different Tyrosinol concentrations were normalized to the control group, and then plotted to calculate the median lethal concentration (IC50).

[0205] The experimental results are shown in Table 7. After Tyrosinol treatment, the growth of the four hepatocellular carcinoma cells was inhibited. Figure 4

[0206] Example 10 Tyraminol inhibits the development of hepatocellular carcinoma in mice

[0207] Objective: To determine the effect of Tyraminol on the development of hepatocellular carcinoma in mice

[0208] Method: 4-week-old Bal / bc nude mice were used. After the mice were anesthetized, the abdominal skin was cut to expose the liver, and HCCLM3 cells were injected into the liver lobule using a syringe. Finally, the liver was returned to its original position, the skin was sutured, and the mice were allowed to wake up. After the mice woke up, they were divided into two groups: a control group and an experimental group (Tyraminol treatment). The experimental group of mice was treated with Tyraminol by gavage every day, with a drug dosage of 500 mg / kg, while the control group was given normal saline by gavage. After 4 weeks of feeding, the mice were sacrificed, and the number of tumors and the maximum diameter of the tumors were counted.

[0209] Results: The results are shown in Table 8. Tyraminol treatment effectively inhibited the number (95.45 vs. 46.33, p < 0.0001) and size (14.79 vs. 6.53, p < 0.0001) of liver tumors in mice.

[0210] Table 8 Effect of Tyraminol on the in situ tumor formation of HCCLM3 cells in the liver

[0211] Number of tumors Tumor diameter (mm) Control group (physiological saline) 95.45 14.79 Experimental group (tyramine) 46.33 6.53

[0212] ​Example 11 Tyrosine content in mouse blood was degraded by tyrosine ammonia lyase treatment

[0213] Purpose of the experiment: To determine the effect of tyrosine ammonia lyase (FjTAL and RgTAL) treatment on the content of tyrosine in mouse liver tissue.

[0214] Experimental method: The change of tyrosine content in the liver tissue of control mice and tyrosine ammonia lyase treated mice was detected by gas chromatography-mass spectrometry method.

[0215] Specifically: 50 mg of liver tissue was taken and added to a tube with 400 μL of methanol-water (v / v = 4:1, containing 0.1% formic acid, containing internal standard succinic acid-2,2,3,3-d4), two small steel balls were added, and after being placed in a refrigerator at -20°C for 2 min, it was ground in a grinder (60HZ, 2 min), then ultrasonic was performed in an ice bath for 10 min, and it was placed in a refrigerator at -20°C for 30 min, centrifuged for 10 min (4°C, 12000 rpm), and 300 μL of supernatant was taken. After taking the supernatant, 300 μL of methanol-water (v / v = 4:1, containing 0.1% formic acid, containing internal standard succinic acid-2,2,3,3-d4) was added to the residue, vortexed for 30 s, ultrasonic was performed in an ice bath for 5 min, centrifuged for 10 min (4°C, 12000 rpm), 300 μL of supernatant was taken, and the total supernatant was 600 μL, which was vortexed and 100 μL of supernatant was taken in a wide buffer tube and dried.

[0216] 80 μL of methoxyamine hydrochloride pyridine solution (15 mg / mL) was added to a glass derivative vial, vortexed for 2 min, and then oximation reaction was performed in a shaking incubator at 37°C (60 min). After the sample was taken out, 80 μL of BSTFA derivatization reagent and 20 μL of n-hexane were added, vortexed for 2 min, and then reacted at 70°C for 60 min. After the sample was taken out, it was placed at room temperature for 30 min, and then GC-MS metabolomics analysis was performed.

[0217] The chromatographic method was: DB-5MS capillary column (30 m x 0.25 mm x 0.25 μm, Agilent J&W Scientific, Folsom, CA, USA), carrier gas was high-purity helium (purity not less than 99.999%), flow rate was 1.2 mL / min, temperature of the injection port was 260°C. The injection volume was 1 μL, non-split injection, and solvent delay was 4 min.

[0218] Programmed temperature: the initial temperature of the column oven is 50℃ for 0.5 min, programmed to 125℃ at 15℃ / min for 2 min, 8℃ / min to 210℃ for 2 min, 11℃ / min to 270℃ for 1 min, 25℃ / min to 305℃ for 3 min. The mass spectrometry method is: electron impact ion source (EI), ion source temperature 300℃, transmission line temperature 280℃, scan mode is selected reaction monitoring scan (SRM), mass scan range: m / z: 40-600.

[0219] The experimental results are shown in Table 1, and the tyrosine content in the liver tissue of the tyrosine ammonia lyase treated mice is significantly reduced compared with the control mice. Figure 6

[0220] From the above description, it can be seen that the above-mentioned embodiments of the present application achieve the following technical effects: the present application first discovers that tyrosine deprivation can significantly inhibit the growth of hepatocellular carcinoma cells, with an inhibition rate of 87.8%, and tyrosine deprivation can also prolong the survival period of mice with hepatocellular carcinoma. Based on the above mechanism, a drug that can reduce tyrosine is developed, which can be used for the treatment of hepatocellular carcinoma. This will provide another new treatment method different from surgery or chemotherapy for patients with hepatocellular carcinoma, and make up for the current lack of effective treatment methods and drugs for hepatocellular carcinoma.

[0221] It should be noted that based on the research results of the present application, the way to reduce the content of tyrosine is not limited to tyrosine ammonia lyase or tyrosol, but can also be achieved by lacking tyrosine from the types of amino acids taken in by hepatocellular carcinoma cells. Therefore, a special medical purpose formula food can also be developed based on this.

[0222] The above only describes the preferred embodiments of the present application and is not intended to limit the present application. For those skilled in the art, the present application can have various modifications and changes. Any modification, equivalent replacement, improvement, etc. made within the spirit and principles of the present application shall be included in the protection scope of the present application.​

Claims

1. Use of tyrosine in the manufacture of a product for treating hepatocellular carcinoma.

2. Use according to claim 1, characterized in that, The mode of treating hepatocellular carcinoma is inhibiting the cell growth of hepatocellular carcinoma.

3. Use according to claim 2, characterized in that, The product comprises an agent for reducing the content of tyrosine in the cells of hepatocellular carcinoma; and / or an agent for blocking the binding of tyrosine and aminoacyl-tRNA synthetase; Preferably, the agent for reducing the content of tyrosine in the cells of hepatocellular carcinoma comprises an agent for reducing the uptake of tyrosine by the cells of hepatocellular carcinoma; Preferably, the agent for blocking the binding of tyrosine and aminoacyl-tRNA synthetase comprises a tyrosine analogue; More preferably, the tyrosine analogue is selected from any one or more of tyrosol, resveratrol or capsaicin; Preferably, the agent for reducing the uptake of tyrosine by the cells of hepatocellular carcinoma comprises a tyrosine-degrading agent and / or a tyrosine-depleted amino acid composition; Preferably, the tyrosine-degrading agent is selected from tyrosine deaminase; More preferably, the tyrosine deaminase is derived from Flavobacterium johnsoniae or Rhodotorula glutinis; Preferably, the tyrosine-depleted amino acid composition comprises essential amino acids and / or non-essential amino acids; Preferably, the essential amino acids are selected from any one or more of methionine, valine, histidine, isoleucine, leucine, lysine, phenylalanine, threonine or tryptophan; Preferably, the non-essential amino acids are selected from any one or more of alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, proline or serine; More preferably, the amino acid composition consists of, in parts by weight: 3.61-3.75 methionine, 4.82-5.0 valine, 3.61-3.75 histidine, 4.82-5.00 isoleucine, 7.23-7.50 leucine, 8.43-8.75 lysine, 1.25-4.82 phenylalanine, 4.82-5.00 threonine, 1.20-1.25 tryptophan, 6.02-6.25 alanine, 6.02-6.25 arginine, 3.01-3.13 asparagine, 6.02-6.25 aspartic acid, 2.41-2.50 cysteine, 18.07-18.75 glutamic acid, 3.01-3.13 glutamine, 6.02-6.25 glycine, 3.01-3.13 proline and 3.01-3.13 serine. More preferably, the amino acid composition consists of, in parts by weight: 3.7-3.75 methionine, 4.94-5.0 valine, 3.70-3.75 histidine, 4.94-5.00 isoleucine, 7.41-7.50 leucine, 8.64-8.75 lysine, 1.25-2.47 phenylalanine, 4.94-5.00 threonine, 1.23-1.25 tryptophan, 6.17-6.25 alanine, 6.17-6.25 arginine, 3.09-3.13 asparagine, 6.17-6.25 aspartic acid, 2.47-2.50 cysteine, 18.52-18.75 glutamic acid, 3.09-3.13 glutamine, 6.17-6.25 glycine, 3.09-3.13 proline, and 3.09-3.13 serine.

4. Use according to claim 2, characterized in that, The application comprises: reducing the content of tyrosine in the cells of the hepatocellular carcinoma and / or blocking the binding of tyrosine and aminoacyl-tRNA synthetase; Preferably, the way of blocking the binding of tyrosine and aminoacyl-tRNA synthetase comprises: making the cells of the hepatocellular carcinoma ingest tyrosine analogues; More preferably, the tyrosine analogues are selected from any one or more of: tyrosol, resveratrol or capsaicin; Preferably, the way of reducing the content of tyrosine in the cells of the hepatocellular carcinoma comprises reducing the ingestion of tyrosine by the cells of the hepatocellular carcinoma; More preferably, the ingestion of tyrosine by the cells of the hepatocellular carcinoma is reduced in the following way: 1) degrading tyrosine in the components to be ingested by the cells of the hepatocellular carcinoma using a tyrosine-degrading agent; and / or 2) making the cells of the hepatocellular carcinoma ingest an amino acid composition lacking tyrosine; Preferably, the tyrosine-degrading agent is selected from tyrosine deaminase; More preferably, the tyrosine deaminase is derived from Flavobacterium johnsoniae or Rhodotorula glutinis; Preferably, the amino acid composition lacking tyrosine comprises essential amino acids and / or non-essential amino acids; wherein the essential amino acids are selected from any one or more of: methionine, valine, histidine, isoleucine, leucine, lysine, phenylalanine, threonine or tryptophan; the non-essential amino acids are selected from any one or more of: alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, proline or serine; Preferably, the amino acid composition consists of, in parts by weight: 3.61-3.75 methionine, 4.82-5.0 valine, 3.61-3.75 histidine, 4.82-5.00 isoleucine, 7.23-7.50 leucine, 8.43-8.75 lysine, 1.25-4.82 phenylalanine, 4.82-5.00 threonine, 1.20-1.25 tryptophan, 6.02-6.25 alanine, 6.02-6.25 arginine, 3.01-3.13 asparagine, 6.02-6.25 aspartic acid, 2.41-2.50 cysteine, 18.07-18.75 glutamic acid, 3.01-3.13 glutamine, 6.02-6.25 glycine, 3.01-3.13 proline and 3.01-3.13 serine; More preferably, the amino acid composition consists of, in parts by weight: 3.7-3.75 methionine, 4.94-5.0 valine, 3.70-3.75 histidine, 4.94-5.00 isoleucine, 7.41-7.50 leucine, 8.64-8.75 lysine, 1.25-2.47 phenylalanine, 4.94-5.00 threonine, 1.23-1.25 tryptophan, 6.17-6.25 alanine, 6.17-6.25 arginine, 3.09-3.13 asparagine, 6.17-6.25 aspartic acid, 2.47-2.50 cysteine, 18.52-18.75 glutamic acid, 3.09-3.13 glutamine, 6.17-6.25 glycine, 3.09-3.13 proline and 3.09-3.13 serine.

5. Use of an agent that reduces the content of tyrosine in cells of hepatocellular carcinoma in the manufacture of a product for the treatment of hepatocellular carcinoma.

6. Use according to claim 5, characterized in that, The agent that reduces the content of tyrosine in cells of hepatocellular carcinoma comprises an agent that reduces the uptake of tyrosine by the cells of the hepatocellular carcinoma; Preferably, the agent that reduces the uptake of tyrosine by the cells of the hepatocellular carcinoma comprises an agent that degrades tyrosine and / or a tyrosine-depleted amino acid composition; Preferably, the agent that degrades tyrosine is selected from tyrosine deaminase; Preferably, the tyrosine deaminase is derived from Flavobacterium johnsoniae or Rhodotorula glutinis; Preferably, the tyrosine-depleted amino acid composition comprises essential amino acids and / or non-essential amino acids; wherein the essential amino acids are selected from any one or more of: methionine, valine, histidine, isoleucine, leucine, lysine, phenylalanine, threonine or tryptophan; the non-essential amino acids are selected from any one or more of: alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, proline or serine; Preferably, the amino acid composition consists of, in parts by weight: 3.61-3.75 methionine, 4.82-5.0 valine, 3.61-3.75 histidine, 4.82-5.00 isoleucine, 7.23-7.50 leucine, 8.43-8.75 lysine, 1.25-4.82 phenylalanine, 4.82-5.00 threonine, 1.20-1.25 tryptophan, 6.02-6.25 alanine, 6.02-6.25 arginine, 3.01-3.13 asparagine, 6.02-6.25 aspartic acid, 2.41-2.50 cysteine, 18.07-18.75 glutamic acid, 3.01-3.13 glutamine, 6.02-6.25 glycine, 3.01-3.13 proline, and 3.01-3.13 serine; More preferably, the amino acid composition consists of, in parts by weight: 3.7-3.75 methionine, 4.94-5.0 valine, 3.70-3.75 histidine, 4.94-5.00 isoleucine, 7.41-7.50 leucine, 8.64-8.75 lysine, 1.25-2.47 phenylalanine, 4.94-5.00 threonine, 1.23-1.25 tryptophan, 6.17-6.25 alanine, 6.17-6.25 arginine, 3.09-3.13 asparagine, 6.17-6.25 aspartic acid, 2.47-2.50 cysteine, 18.52-18.75 glutamic acid, 3.09-3.13 glutamine, 6.17-6.25 glycine, 3.09-3.13 proline, and 3.09-3.13 serine.

7. Use of an agent that blocks the binding of tyrosine to aminoacyl-tRNA synthetase in the manufacture of a medicament for the treatment of hepatocellular carcinoma.

8. Use according to claim 7, characterized in that, The agent that blocks the binding of tyrosine to aminoacyl-tRNA synthetase comprises a tyrosine analog; Preferably, the tyrosine analog is selected from any one or more of: tyrosol, resveratrol, or capsaicin.

9. A medicament for treating hepatocellular carcinoma, characterized by, The medicament comprises an agent that reduces the content of tyrosine in the cells of the hepatocellular carcinoma and / or an agent that blocks the binding of tyrosine to aminoacyl-tRNA synthetase; Preferably, the agent that reduces the content of tyrosine in the cells of the hepatocellular carcinoma comprises an agent that reduces the uptake of tyrosine by the cells of the hepatocellular carcinoma; More preferably, the agent that reduces the uptake of tyrosine by the cells of the hepatocellular carcinoma comprises an agent that degrades tyrosine and / or a tyrosine-depleted amino acid composition; Further preferably, the agent that degrades tyrosine is selected from tyrosine deaminase; Preferably, the tyrosine deaminase is derived from Flavobacterium johnsoniae or Rhodotorula glutinis; Further preferably, the tyrosine-depleted amino acid composition comprises essential amino acids and / or non-essential amino acids; wherein the essential amino acids are selected from any one or more of: methionine, valine, histidine, isoleucine, leucine, lysine, phenylalanine, threonine, or tryptophan; the non-essential amino acids are selected from any one or more of: alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, proline, or serine; more preferably, the amino acid composition consists of, in parts by weight: 3.7-3.75 methionine, 4.94-5.0 valine, 3.70-3.75 histidine, 4.94-5.00 isoleucine, 7.41-7.50 leucine, 8.64-8.75 lysine, 1.25-2.47 phenylalanine, 4.94-5.00 threonine, 1.23-1.25 tryptophan, 6.17-6.25 alanine, 6.17-6.25 arginine, 3.09-3.13 asparagine, 6.17-6.25 aspartic acid, 2.47-2.50 cysteine, 18.52-18.75 glutamic acid, 3.09-3.13 glutamine, 6.17-6.25 glycine, 3.09-3.13 proline, and 3.09-3.13 serine; more preferably, the amino acid composition consists of, in parts by weight: 3.7-3.75 methionine, 4.94-5.0 valine, 3.70-3.75 histidine, 4.94-5.00 isoleucine, 7.41-7.50 leucine, 8.64-8.75 lysine, 1.25-2.47 phenylalanine, 4.94-5.00 threonine, 1.23-1.25 tryptophan, 6.17-6.25 alanine, 6.17-6.25 arginine, 3.09-3.13 asparagine, 6.17-6.25 aspartic acid, 2.47-2.50 cysteine, 18.52-18.75 glutamic acid, 3.09-3.13 glutamine, 6.17-6.25 glycine, 3.09-3.13 proline, and 3.09-3.13 serine; preferably, the agent that blocks the binding of tyrosine and aminoacyl-tRNA synthetase comprises a tyrosine analog; more preferably, the tyrosine analog is selected from any one or more of: tyrosol, resveratrol, or capsaicin.

10. An amino acid composition, characterized in that, the amino acid composition consists of, in parts by weight: 3.61-3.75 methionine, 4.82-5.0 valine, 3.61-3.75 histidine, 4.82-5.00 isoleucine, 7.23-7.50 leucine, 8.43-8.75 lysine, 1.25-4.82 phenylalanine, 4.82-5.00 threonine, 1.20-1.25 tryptophan, 6.02-6.25 alanine, 6.02-6.25 arginine, 3.01-3.13 asparagine, 6.02-6.25 aspartic acid, 2.41-2.50 cysteine, 18.07-18.75 glutamic acid, 3.01-3.13 glutamine, 6.02-6.25 glycine, 3.01-3.13 proline, and 3.01-3.13 serine; the amino acid composition consists of, in parts by weight: 3.61-3.75 methionine, 4.82-5.0 valine, 3.61-3.75 histidine, 4.82-5.00 isoleucine, 7.23-7.50 leucine, 8.43-8.75 lysine, 1.25-4.82 phenylalanine, 4.82-5.00 threonine, 1.20-1.25 tryptophan, 6.02-6.25 alanine, 6.02-6.25 arginine, 3.01-3.13 asparagine, 6.02-6.25 aspartic acid, 2.41-2.50 cysteine, 18.07-18.75 glutamic acid, 3.01-3.13 glutamine, 6.02-6.25 glycine, 3.01-3.13 proline, and 3.01-3.13 serine; the amino acid composition consists of, in parts by weight: 3.61-3.75 methionine, 4.82-5.0 valine, 3.61-3.75 histidine, 4.82-5.00 isoleucine, 7.23-7.50 leucine, 8.43-8.75 lysine, 1.25-4.82 phenylalanine, 4.82-5.00 threonine, 1.20-1.25 tryptophan, 6.02-6.25 alanine, 6.02-6.25 arginine, 3.01-3.13 asparagine, 6.02-6.25 aspartic acid, 2.41-2.50 cysteine, 18.07-18.75 glutamic acid, 3.01-3.13 glutamine, 6.02-6.25 glycine, 3.01-3.13 proline, and 3.01-3.13 serine;

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