WO2020125551A1

WO2020125551A1 - Application of thioguanine in preparation of drugs for treatment, improvement or prevention of tumors

Info

Publication number: WO2020125551A1
Application number: PCT/CN2019/125154
Authority: WO
Inventors: 朱威; 潘武广
Original assignee: 广州君赫生物科技有限公司
Priority date: 2018-12-19
Filing date: 2019-12-13
Publication date: 2020-06-25

Abstract

Provided by the present invention is an application of thioguanine in the preparation of drugs for the treatment, improvement or prevention of tumors; thioguanine has a significant inhibitory effect on cancer having abnormally high expression of PAICS gene.

Description

Thioguanine is used in the preparation of drugs for treating, improving or preventing tumors

Technical field

The invention relates to the new application of thioguanine, in particular to the application of thioguanine in the preparation of medicines for treatment, improvement or prevention of tumors.

Background technique

Tumors, especially malignant tumors, seriously threaten people's health. The pathogenesis of tumors is diverse, because different tumors have different mechanisms, resulting in different responses to different compounds. Chemotherapy drugs have a certain therapeutic effect on most tumors, mainly because they have a broad-spectrum lethal effect on cells. Compounds that do not have a general lethal effect on cells are often only effective for specific tumors, but not for other tumors, and even have the risk of promoting tumor progression. Therefore, its use is severely restricted.

An important hallmark of cancer cells is metabolic reprogramming, including increased glucose uptake and non-oxygen-dependent lactic acid fermentation, which is also often referred to as the Warburg effect [1,2]. This reprogramming is necessary for the growth and survival of the tumor, especially under pressure conditions such as external hypoxia. However, the important molecular mechanism and mode of action for the correlation between tumor cell metabolism reorganization and tumor rapid proliferation, differentiation, migration, etc. are still unclear.

Pyruvate kinase isoenzyme 2 (Pyruvate kinase isoform M2, PKM2), as an important enzyme in the metabolic process, is highly expressed in rapid proliferation and most tumor cells, and has a huge impact on tumor cell metabolism and growth [3, 4]. In addition, various pharmacological agents targeting PKM2 enzyme activity affect cell growth and proliferation [5, 6], which also suggests that by targeting PKM2 enzyme activity as a representative, further changes in the way of tumor metabolism have become a new approach to cancer treatment [ 7].

Purine anabolic metabolism is a ubiquitous and very important biological metabolism of organisms. Its products AMP and GMP not only provide raw materials for the biosynthesis of DNA and RNA in the organism, but also provide many key coenzymes (NAD, NADP, FAD and CoA) in the body. ), signaling molecules (such as cAMP) and important energy molecules ATP provide the necessary purine bases for their synthesis. It can be seen that purine anabolism is at the core of the entire metabolic network. Purine synthesis includes de novo purification (synthesis) and salvage pathway (salvage pathway).

In the metabolic pathway of de novo adenine synthesis, adenyl succinate lyase (hereinafter referred to as ADSL enzyme) is mainly involved in the reaction of catalyzing SAICAR to form AICAR and S-AMP to generate AMP [Spiegel, EK, Colman, RF, and Patterson , D. (2006). Adenylosuccinate Lyase definition. Mol Genet Metab 89, 19-31. Clamadieu, C., Cottin, X., Rousselle, C., and Claris, O. (2008). Adenylosuccinate Lyase definition: an unusual cause Neonatal Seizure. Arch Pediatr 15, 135-138. Castro, M., Perez-Cerda, C., Merinero, B., Garcia, MJ, Bernar, J., Gil Nagel, A., Torres, J., Bermudez, M., Garavito, P., Marie, S., et.al. (2002). Screening for adenylosuccinatelyase definition: clinical, biochemical and molecularfindings in four patients. Neuropediatrics 33, 186-189.].

Abnormal metabolic enzymes in the de novo adenine anabolic pathway in the human body often lead to the intermediate harmful metabolite 5-amino-4-succinic acid formamide imidazole ribonucleotide (succinyl-5-aminoimidazole-4-carboxamide-1- The accumulation of ribose-50-phosphate (SAICAR) is clinically manifested as autism, epilepsy, hypotonia, dysplasia and other symptoms [8-10]. SAICAR synthase is encoded by the gene PAICS (phosphoribosylaminoimidazolesuccinocarboxamide synthase) and is responsible for SAICAR synthesis in the body. Related studies have reported that PAICS is highly expressed in acute lymphocytic leukemia, lung cancer, glioma, prostate cancer, and rectal cancer, and can be used as a prognostic marker for stage III colorectal cancer [11-13]. Recent studies have found that SAICAR is highly accumulated under glucose-limited conditions, which changes the energy levels, sugar uptake and lactic acid production in tumor cells, and these phenomena have not occurred in adult epidermal cells and lung fibroblasts [14, 15]. SAICAR can induce the enzymatic activity of PKM2 and promote the survival of tumor cells [14], and the combination of SAICAR-PKM2 can induce the phosphorylation of Erk1/2, high concentration of SAICAR can also induce the up-regulated expression of oncogene myc [15], these SAICAR, which accumulates abnormally in the de novo anabolic pathway of adenine, promotes the proliferation and survival of tumor cells.

Aminoimidazole succinylcarbamoyl nucleotide synthetase/aminoimidazole nucleotide carboxylase, namely PAICS (phosphoribosylaminoimidazole succinocarboxamide/synthetase/phosphoribosylaminoimidazole carboxylase) is an important bifunctional enzyme in the de novo synthesis of purine, which has SAICAR synthesis Enzymes (4-(N-succinylcarboxamide)-5-aminoimidazole ribonucleotide synthetase, SAICARs) and AIR carboxylase (5-aminoimidazole ribonucleotide carboxylase, AIRc) function, catalyze the de novo synthesis of purines in the sixth and seventh reactions, in which A key reaction process of

Therefore, tumors with abnormally high expression of PAICS are often accompanied by the accumulation of harmful metabolites SAICAR, SAICAr or S-Ado, and the research on the inhibition of PAICS expression or its enzymatic activity will become a new method for tumor treatment. It is of great significance to develop or screen out compounds that can effectively inhibit PAICS activity.

DB00352 (https://www.drugbank.ca/drugs/DB00352), common name, Tioguanine, structural formula:

Chinese name: thioguanine. The main uses of thioguanine are as follows:

1. It can cause obvious transient hyperplasia of white blood cells. It is clinically used for the treatment of leukopenia and non-specific thrombocytopenia caused by various radioactivity or drugs. It is also used for acute and chronic hepatitis. Adverse reactions include dizziness, palpitations, vomiting, chest tightness, and nausea.

2. Antitumor drugs. It can relieve acute leukemia and chronic myeloid leukemia. Large doses have a good effect on choriocarcinoma.

3. Protein kinase activator. It is a derivative of adenosine cyclophosphate, its function and use are the same. It can also be used for myocarditis, cardiogenic shock, and submental hemorrhage after surgery. When the dosage is large, there may be drowsiness, dizziness, vertigo, fatigue, loss of appetite, nausea, vomiting, etc.

4. Antitumor drugs. It is a kind of purine antimetabolite. It is a homologue of DNA bases and can be incorporated into DNA molecules as a wrong base, thereby affecting the synthesis and function of DNA. It mainly acts on S phase and is a cell cycle specific drug. For the treatment of various types of leukemia, acute leukemia is more commonly used.

5. For the prevention and treatment of coronary atherosclerosis and the treatment of hepatitis. It is also used for various metabolic auxiliary treatments such as leukopenia, primary thrombocytopenic purpura, acute anuria caused by chronic renal insufficiency, nephrotic syndrome, and uremia.

Its pharmacological action is similar to mercaptopurine, and it is only active after it is converted into thioguanine uridine (6-TGRP) in the body. Finally, it is converted into deoxyguanine nucleotides, which interferes with DNA function and produces anti-cancer effects. It is an S-phase specific antitumor drug, which has a delaying effect on the S/G2 boundary. Commonly used purine metabolism antagonists that inhibit purine synthesis pathways are cell cycle-specific drugs that are most sensitive to cells in S phase. In addition to inhibiting the synthesis of cellular DNA, they also have a slight inhibitory effect on RNA synthesis. This product is an analog of guanine. It must be converted from phosphoribosyltransferase to 6-TG ribonucleotide in the human body to be active. The action of this product is similar to that of mercaptopurine. In addition, 6-TG ribonucleotide Through the inhibition of guanylate kinase, the phosphorylation of guanosine monophosphate (GMP) to guanosine diphosphate (GDP) can be prevented. After being metabolized to deoxyribose triphosphate, this product can be incorporated into DNA, thus further inhibiting the biosynthesis of nucleic acids, mercaptopurine has no such effect. This product is effectively cross-resistant to mercaptopurine, and combined with cytarabine and other drugs can improve the efficacy.

Existing studies have shown that thioguanine has only a good effect in some tumors and has certain side effects. How to expand the therapeutic scope of thioguanine has very practical significance.

references:

1. Hanahan, D. and R. A. Weinberg, Hallmarks of cancer: the next generation. Cell, 2011.144(5): p.646-74.

2. Hsu, P.P. and D. M. Sabatini, Cancer cell metabolism: Warburg and beyond. Cell, 2008.134(5): p.703-7.

3. Christofk, H.R., et.al., The M2 splice isoform of pyruvatekinase is important for cancer metabolicism and tumorour growth. Nature, 2008.452(7184): p.230-3.

4.Wolf, A., et.al., Hexokinase 2 isis a key, mediator of aerobic glycolysis, tumors, tumor, growth, human glioblastoma, multiforme. J Exp, Med, 2011.208(2): p.313-26.

5.Chen, J., et.al., Shikonin and its analogs inhibit cell glycolysis by targeting tumor tumor-M2.Oncogene, 2011.30(42): p.4297-306.

6. Anastasiou, D., et.al., Pyruvatekinase M2activatorspromotetetramerformation anduppresstumorigenesis.NatChemBiol, 2012.8(10): p.839-47.

7. Vander Heiden, M.G., Exploiting tumor metabolism: challenges for clinical translation. J Clin Invest, 2013.123(9): p.3648-51.

8. Ciardo, F., C. Salerno, and P. Curatolo, Neurologic aspects of of adenylosuccinately definition. J Child Child Neurol, 2001.16(5): p.301-8.

9. Gitiaux, C., et.al., Misleading behavioural phenotype with adenylosuccinate lyase definition. Eur JHum Genet, 2009.17(1): p.133-6.

10. Mierzewska, H., et.al., Severeencephalopathy withbrain atrophy and hypomyelination due to nynylosuccinate lyasedeficiency--MRI, clinical, biochemical and neuropathologicalfindingsofofPolishpatients.FoliaNeuropathol: 2009.47 (2009.47) (2009.47) (47.20): .

11.Eissmann, M., et.al., A functional functional survival screen-of tumor-derived cDNA libraries designed for anti-apoptotic mammalian oncogenes.PLoS One, 2013.8(5): p.e64873.

12.Goswami, M.T., et.al., Role and regulation of coordinately expressed, novopurine, biosynthetic, enzymes, PPAT, PAICS, cancer, Oncotarget, 2015.6(27): p.23445-61.

13. Chakravarthi, B.V., et.al., Expression and Role of PAICS, aDe Novo Purine Biosynthetic Gene Prostate Cancer: Prostate, 2017.77(1): p.10-21.

14. Keller, K.E., I.S. Tan, and Y.S. Lee, SAICAR stimulations pyruvate skinase isoform M2 and promotes cancer cell survival survival in glucose-limited conditions. Science, 2012.338 (6110): p.1069-72.

15. Keller, K.E., et.al., SAICAR induces protein proteinase activity PKM2 that is necessary for proliferative signaling of cancer cells. Mol Cell, 2014.53(5): p.700-9.

Summary of the invention

The purpose of the present invention is to provide the application of thioguanine in the preparation of drugs for treating, improving or preventing tumors.

Summary of the invention:

The first aspect of the present invention provides:

Thioguanine and its pharmaceutically acceptable derivatives, prodrugs and active metabolites are used in the preparation of drugs for treatment, improvement or prevention of tumors,

The tumor has any of the following characteristics:

The amount of SAICAR is higher than normal, there is excessive accumulation of SAICAR, SAICAr or S-Ado;

Warburg effect, high expression of oncogene myc, high expression of PAICS, correlation with Erk1/2 and high expression of PKM2 gene.

In some examples, the tumor is selected from the group consisting of lung cancer, breast cancer, colon cancer, rectal cancer, prostate cancer, bladder cancer, cervical cancer, liver cancer, cholangiocarcinoma, esophageal cancer, polyglioblastoma, head and neck squamous cells Cancer, pancreatic cancer, gastric cancer, endometrial cancer, leukemia.

In some examples, the tumor has any of the following characteristics:

PAICS high expression.

In some examples, the drug also includes at least one compound and/or agent that has a therapeutic effect on the tumor.

The second aspect of the present invention provides:

Compounds used to treat, improve or prevent tumor drugs,

The compound is thioguanine and its pharmaceutically acceptable derivatives, prodrugs and active metabolites;

The tumor has any of the following characteristics:

In some examples, the tumor has any of the following characteristics:

PAICS high expression.

The third aspect of the present invention provides:

A composition for treating, improving or preventing tumors,

The active ingredients of the composition include: thioguanine and its pharmaceutically acceptable derivatives, prodrugs and active metabolites;

The tumor has any of the following characteristics:

In some examples, the tumor has any of the following characteristics:

PAICS high expression.

In some examples, the composition also includes at least one compound and/or agent that has a therapeutic effect on the tumor.

The fourth aspect of the present invention provides:

A method for treating, ameliorating or preventing tumors, which includes administering to a patient a therapeutic amount, an improved amount or a preventive amount of thioguanine and its pharmaceutically acceptable derivatives, prodrugs, and active metabolites,

The tumor has any of the following characteristics:

In some examples, the tumor has any of the following characteristics:

PAICS high expression.

In some examples, it also includes administering at least one compound and/or agent that has a therapeutic effect on the tumor.

The beneficial effects of the invention are:

The inventor discovered during the experiment that thioguanine has a good inhibitory effect on some tumor cells, while the inhibitory effect on some tumor cells is significantly weakened. This characteristic of thioguanine is different from most chemotherapy drugs and has its unique and unknown pattern.

To further determine the cause, the inventors based on the TCGA database (The Cancer Genome Atlas) about 10,000 samples of gene expression bioinformatics analysis, combined with further cell biology experiments, found and confirmed that thioguanine on PAICS gene abnormalities Highly expressed cancer has a significant inhibitory effect (as shown in Figure 1, Table 1, Table 2), can treat lung cancer, breast cancer, colon cancer, rectal cancer, prostate cancer, bladder cancer, cervical cancer, liver cancer, bile duct cancer, Cancers with abnormally high expression of PAICS genes, such as esophageal cancer, polymorphic glioblastoma, squamous cell carcinoma of the head and neck, pancreatic cancer, gastric cancer, endometrial cancer, and leukemia. Cancers such as paraganglioma and non-abnormally high expression of PAICS genes have no significant therapeutic effect, proving that thioguanine can be developed as a new targeted anticancer drug for cancers with abnormally high expression of oncogene PAICS.

BRIEF DESCRIPTION

Figure 1 shows the PAICS gene expression of different tumors.

detailed description

After cell biology experiments and gene expression bioinformatics analysis of about 10,000 samples of TCGA database (The Cancer Genome Atlas), it was found that thioguanine has a significant inhibitory effect on cancers with abnormally high expression of PAICS gene (Figure 1 , Table 1, Table 2), can treat lung cancer, breast cancer, colon cancer, rectal cancer, prostate cancer, bladder cancer, cervical cancer, liver cancer, cholangiocarcinoma, esophageal cancer, multiform glioblastoma, head and neck scales Cancers with abnormally high expression of PAICS genes such as squamous cell carcinoma, pancreatic cancer, gastric cancer, endometrial cancer, and leukemia, and those with abnormally high expression of PAICS genes such as renal cancer, cutaneous melanoma, pheochromocytoma, and paraganglioma Cancer has no significant therapeutic effect, proving that thioguanine can be developed as a new targeted anti-cancer drug for cancers with abnormally high expression of oncogene PAICS.

Table 1. Anticancer effect of thioguanine and the targeting correlation of high expression of PAICS oncogene

Table 2. Gene expression bioinformatics analysis table of about 10,000 sample data of TCGA database

Animal experiment:

Both ADSL gene-deficient nematodes and ADSL-deficient transgenic mice have excessive accumulation of SAICAR, and their phenotype is similar to the high expression of PAICS gene.

After thioguanine treatment, the phenotype of the ADSL gene-deficient nematode animal model can be restored to normal.

After the administration of thioguanine in ADSL-deficient transgenic mice, experiments have shown that the accumulation of toxic intermediate metabolites such as SAICAR, SAICAr, S-Ado, etc. has decreased significantly, and ADSL has been significantly improved. This shows that it has a significant therapeutic effect on tumors with high expression of PAICS genes caused or promoted by the accumulation of toxic intermediate metabolites such as SAICAR, SAICAr, and S-Ado.

Similarly, thioguanine pharmaceutically acceptable derivatives, prodrugs and active metabolites have significant effects on tumors with high expression of PAICS genes caused or promoted by the accumulation of toxic intermediate metabolites such as SAICAR, SAICAr, S-Ado, etc. Therapeutic effect.

Claims

Thioguanine and its pharmaceutically acceptable derivatives, prodrugs and active metabolites are used in the preparation of drugs for treatment, improvement or prevention of tumors, and are characterized by:

The tumor has any of the following characteristics:

The amount of SAICAR is higher than normal, there is excessive accumulation of SAICAR, SAICAr or S-Ado;

Warburg effect, high expression of oncogene myc, high expression of PAICS, correlation with Erk1/2 and high expression of PKM2 gene.
The use according to claim 1, characterized in that the tumor is selected from the group consisting of lung cancer, breast cancer, colon cancer, rectal cancer, prostate cancer, bladder cancer, cervical cancer, liver cancer, cholangiocarcinoma, esophageal cancer, and multi-forming gel Cytoblastoma, squamous cell carcinoma of the head and neck, pancreatic cancer, gastric cancer, endometrial cancer, leukemia.
The application according to claim 2, wherein the tumor has any of the following characteristics:

The amount of SAICAR is higher than normal, there is excessive accumulation of SAICAR, SAICAr or S-Ado;

PAICS high expression.
The use according to claims 1 to 3, characterized in that the medicine further comprises at least one compound and/or agent having a therapeutic effect on tumors.
Compounds for the treatment, improvement or prevention of tumor drugs, characterized by:

The compound is thioguanine and its pharmaceutically acceptable derivatives, prodrugs and active metabolites;

The tumor has any of the following characteristics:

The amount of SAICAR is higher than normal, there is excessive accumulation of SAICAR, SAICAr or S-Ado;

Warburg effect, high expression of oncogene myc, high expression of PAICS, correlation with Erk1/2 and high expression of PKM2 gene.
The compound according to claim 5, wherein the tumor is selected from the group consisting of lung cancer, breast cancer, colon cancer, rectal cancer, prostate cancer, bladder cancer, cervical cancer, liver cancer, cholangiocarcinoma, esophageal cancer, and multi-forming gel Cytoblastoma, squamous cell carcinoma of the head and neck, pancreatic cancer, gastric cancer, endometrial cancer, leukemia.
The compound according to claim 6, wherein the tumor has any of the following characteristics:

The amount of SAICAR is higher than normal, there is excessive accumulation of SAICAR, SAICAr or S-Ado;

PAICS high expression.
A composition for treating, improving or preventing tumors, characterized by:

The active ingredients of the composition include: thioguanine and its pharmaceutically acceptable derivatives, prodrugs and active metabolites;

The tumor has any of the following characteristics:

The amount of SAICAR is higher than normal, there is excessive accumulation of SAICAR, SAICAr or S-Ado;

Warburg effect, high expression of oncogene myc, high expression of PAICS, correlation with Erk1/2 and high expression of PKM2 gene.
The composition according to claim 8, wherein the tumor is selected from lung cancer, breast cancer, colon cancer, rectal cancer, prostate cancer, bladder cancer, cervical cancer, liver cancer, cholangiocarcinoma, esophageal cancer, polymorphism Glioma, head and neck squamous cell carcinoma, pancreatic cancer, gastric cancer, endometrial cancer, leukemia.
The composition according to claim 9, wherein the tumor has any of the following characteristics:

The amount of SAICAR is higher than normal, there is excessive accumulation of SAICAR, SAICAr or S-Ado;

PAICS high expression.
The composition according to claims 8 to 10, characterized in that the composition further comprises at least one compound and/or agent having a therapeutic effect on tumors.
A method for treating, ameliorating or preventing tumors, including administering to a patient a therapeutic amount, an improved amount or a preventive amount of thioguanine and its pharmaceutically acceptable derivatives, prodrugs, and active metabolites, characterized in that :

The tumor has any of the following characteristics:

The amount of SAICAR is higher than normal, there is excessive accumulation of SAICAR, SAICAr or S-Ado;

Warburg effect, high expression of oncogene myc, high expression of PAICS, correlation with Erk1/2 and high expression of PKM2 gene.
The method according to claim 12, wherein the tumor is selected from the group consisting of lung cancer, breast cancer, colon cancer, rectal cancer, prostate cancer, bladder cancer, cervical cancer, liver cancer, cholangiocarcinoma, esophageal cancer, and multi-forming gel Cytoblastoma, squamous cell carcinoma of the head and neck, pancreatic cancer, gastric cancer, endometrial cancer, leukemia.
The method according to claim 13, wherein the tumor has any of the following characteristics:

The amount of SAICAR is higher than normal, there is excessive accumulation of SAICAR, SAICAr or S-Ado;

PAICS high expression.
The method according to claims 12-14, further comprising administering at least one compound and/or agent having a therapeutic effect on the tumor.