Use of tetrahydroisoquinoline compound in preparation of Anti-leukemia drug

Abstract

Disclosed is use of a tetrahydroisoquinoline compound in the preparation of an anti-leukemia drug. The tetrahydroisoquinoline compound has a significant inhibitory effect on leukemia cells and low toxicity to organisms. This compound expands the range of drugs for clinical treatment of leukemia, and is suitable for mass production.

Description

Application of a tetrahydroisoquinoline compound in the preparation of anti-leukemia drugs Technical Field

[0001] This invention relates to the field of biomedical technology, and in particular to the application of a tetrahydroisoquinoline compound in the preparation of anti-leukemia drugs. Background Technology

[0002] Malignant tumors, a major health challenge threatening human life, continue to be the leading cause of death worldwide. In recent years, the situation of cancer in my country has been particularly severe, with both the overall incidence and mortality rates showing a continuous upward trend. This trend not only poses a huge threat to individual health but also places a heavy burden on the entire society. Authoritative statistics show that the annual incidence rate of malignant tumors in my country is as high as 3.9%, while the mortality rate follows closely behind, rising by 2.5%. Behind these figures lies the heavy blow suffered by countless families due to cancer and the enormous pressure on social medical resources.

[0003] Despite advancements in cancer treatment and improved medical standards thanks to technological progress, resulting in better survival rates for common types of malignant tumors, malignant tumors remain a significant threat to human health. Among these, hematological malignancies, as a special type of malignant tumor, receive particular attention due to their wide age range of onset and the difficulty in treatment.

[0004] Hematologic malignancies, as the name suggests, are a group of diseases caused by the cancerous transformation and malignant proliferation of cells in the hematopoietic system. These cancers can occur at a very wide age range, from infants to the elderly, making them one of the cancers with the widest age of onset. The main characteristic of hematologic malignancies is the abnormality in the quantity and quality of white blood cells and their immature cells (precursor cells or leukemia cells) in the blood and bone marrow. These abnormal cells suppress the normal hematopoietic function of the bone marrow and infiltrate tissues and organs such as the liver, spleen, and lymph nodes, leading to symptoms such as bleeding, bruising, fatigue, and an increased risk of infection.

[0005] The classification of hematologic malignancies is highly complex, encompassing various types of leukemia, lymphoma, and others. These diseases differ in their pathogenesis, clinical manifestations, and treatments. Furthermore, most hematologic malignancies are associated with genetic alterations, making diagnosis and treatment even more challenging. For example, acute myeloid leukemia (AML) is a clonal hematologic malignancy caused by abnormal development of hematopoietic stem cells or progenitor cells, and its incidence increases with age. Acute lymphoblastic leukemia (ALL), on the other hand, is caused by the excessive proliferation of lymphoblasts due to unexplained maturation disorders, which suppresses normal bone marrow hematopoietic tissue and leads to infiltration of peripheral blood vessels, bone marrow, systemic lymph nodes, spleen, and liver. ALL is particularly common in adults.

[0006] Currently, the main clinical treatment for leukemia involves a combination of hematopoietic stem cell transplantation and chemotherapy. However, while these methods can improve patient survival rates to some extent, many patients still experience relapse or lack of treatment response. Furthermore, the variety of drugs available for treating leukemia remains limited, failing to meet the growing clinical demand. Therefore, there is an urgent need to develop more anti-leukemia drugs, especially those with higher targeting efficiency and stronger ability to penetrate cancer cell membranes, providing more options for medical research and clinical application.

[0007] Summary of the Invention

[0008] To address the shortcomings of existing technologies, this invention proposes the application of tetrahydroisoquinoline compounds in the preparation of anti-leukemia drugs.

[0009] This invention provides the application of a tetrahydroisoquinoline compound in the preparation of an anti-leukemia drug, wherein the tetrahydroisoquinoline compound has the structure of formula (I):

[0010] in, It is a substituted amide group formed by the carboxylic acid and amino group on an amino acid, or R1 is hydrogen or C. 1～12 Alkyl, C 1～12 Alkoxy, C 1～12 Alkyl group, C 1～12 Any substituted amide group among alkyl ester groups;

[0011] R2 is X is any one of C or N atoms, R 21 for R 211 It may be mono- or poly-substituted, selected from one or more of hydrogen, halogen, unsubstituted or substituted piperazine, indole, and morpholine, wherein the substituent of the substituted piperazine is C. 1～6 Alkyl, halogen, C 1～6 Any one of alkoxy, hydroxyl, or amino groups;

[0012] The tetrahydroisoquinoline compounds of this invention can, on the one hand, target and inhibit the interaction between nuclear receptor coactivator 4 and ferritin heavy chain, inhibit ferrophagy, thereby inhibiting the proliferation of tumor cells, especially showing significant inhibitory effects on leukemia cells, achieving a therapeutic effect on leukemia; on the other hand, the inventors discovered that the amide group in the tetrahydroisoquinoline compounds... This can improve the drug's targeting ability and its ability to penetrate cancer cell membranes, thereby enhancing the compound's activity. Since cancer cells typically carry a weak negative charge on their surface, positively charged amide molecules are more easily reached and absorbed, resulting in higher anti-cancer effects. Therefore, the tetrahydroisoquinoline compounds of this invention can provide more options for medical research and clinical applications.

[0013] Furthermore, the aforementioned The substituted amide group is formed by the carboxylic acid and amino group of any one of the amino acids selected from alanine, phenylalanine, L-alanine, cysteine, selenocysteine, aspartic acid, asparagine, glutamic acid, glutamine, glycine, histidine, leucine, isoleucine, lysine, pyrrolidone, methionine, proline, arginine, serine, threonine, valine, tryptophan, and tyrosine, or R1 is hydrogen or carbon. 1～12 Alkyl, C 1～12 Any substituted amide group among alkoxy groups;

[0014] The R 211 The substituted piperazine has a C substituent. 1～3 alkyl.

[0015] Furthermore, the aforementioned The substituted amide group is formed by the carboxylic acid and amino group of any one of the amino acids selected from alanine, L-alanine, aspartic acid, asparagine, glutamic acid, and glutamine, or R1 is C. 1～12 Alkyl, C 1～12 Any substituted amide group of alkoxy groups.

[0016] The following is an exemplary synthetic route for a tetrahydroisoquinoline compound, where R2 of example compound 1 is... X is a C atom, R 21 for R 211 For monosubstituted piperazines, the substituent is methyl:

[0017] The following is a method for preparing example compound 1, including the following steps:

[0018] Compound 2 (0.9-1.1 mmol), TBTU (0.9-1.1 mmol), and K2CO3 (1.4-1.6 mmol) were added sequentially to a round-bottom flask. Dry N,N-dimethylformamide (9-11 mL) was added, and the mixture was stirred at room temperature for 50-70 min. Then, compound 3 (0.8-1.2 mmol) was added, and the mixture was magnetically stirred at 60 °C for 11-13 h. The mixture was then extracted with ethyl acetate (3 × 150 mL, i.e., three extractions with 150 mL of ethyl acetate). The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The solid obtained by rotary evaporation was purified by column chromatography to obtain a yellow solid, yielding compound 4 (yield 61%-83%).

[0019] Compound 4 (0.9-1.1 mmol) was added to 4-6 mL of methanol, followed by 1-2 mL of TFA. After stirring at room temperature for 2.5-3.5 hours, saturated sodium carbonate solution was added to adjust the pH to around 8. The mixture was then extracted with ethyl acetate (3 × 150 mL). The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain crude compound 5. Compound 6 (1.9-1.1 mmol) and DBU (1.1-1.3 mmol) were then added and dissolved in N,N-dimethylformamide (9-11 mL). The mixture was magnetically stirred at 60 °C for 7-9 hours. The mixture was then extracted with ethyl acetate (3 × 150 mL). The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The solid obtained by rotary evaporation was purified by column chromatography to obtain a yellow solid, yielding compound 7 (yield 59%-77%).

[0020] Compound 7 (0.9-1.1 mmol), compound 8 (0.9-1.1 mmol), cesium carbonate (1.1-1.3 mmol), and Pd(OAc)2 (0.9-1.1 mmol) were added to 9-11 mL of methanol and stirred at 60 °C for 7.5-8.5 h. After the reaction was completed, the solid was removed by filtration. The solid was washed with methanol (3 × 10 mL, i.e., washed three times with 10 mL of methanol). The filtrate was then evaporated to dryness and purified by column chromatography to obtain a yellow solid, which was compound 1.

[0021] Furthermore, the tetrahydroisoquinoline compound has any one of the following structural formulas:

[0022] Preferred

[0023] Furthermore, the tetrahydroisoquinoline compound may be used in any of its pharmaceutically acceptable salts, solvates, or chiral isomers.

[0024] Furthermore, the pharmaceutically acceptable salt is obtained by reacting a tetrahydroisoquinoline compound with an inorganic or organic acid.

[0025] Further, the organic acid is any one of citric acid, acetic acid, oxalic acid, isonicotinic acid, lactic acid, salicylic acid, acidic citric acid, tartaric acid, oleic acid, tannic acid, pantothenic acid, tartaric acid, ascorbic acid, succinic acid, maleic acid, gentic acid, fumaric acid, gluconic acid, glucuronic acid, glycolic acid, formic acid, benzoic acid, glutamic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, or dihydroxynaphthyl acid.

[0026] Furthermore, the inorganic acid is any one of hydrochloric acid, phosphoric acid, sulfuric acid, or nitric acid.

[0027] Furthermore, the drug is any one of an oral preparation, an injection, a topical preparation, or an inhaler.

[0028] Furthermore, the oral medication is any one of capsules, tablets, pills, or granules; the inhalant is a spray.

[0029] In summary, compared with the prior art, the present invention achieves the following technical effects:

[0030] (1) The tetrahydroisoquinoline compounds provided by this invention can significantly inhibit the proliferation of leukemia cells;

[0031] (2) The tetrahydroisoquinoline compounds provided by this invention have high targeting ability and stronger ability to penetrate cancer cell membranes, which can enhance compound activity and have high anti-cancer effect.

[0032] (3) The preparation process of tetrahydroisoquinoline compounds provided by the present invention is simple and easy to produce on a large scale. Attached Figure Description

[0033] To more clearly illustrate the technical solutions of the embodiments of the present invention, the accompanying drawings used in the embodiments will be briefly introduced below. It should be understood that the following drawings only show some embodiments of the present invention and should not be regarded as a limitation on the scope. For those skilled in the art, other related drawings can be obtained based on these drawings without creative effort.

[0034] Figure 1 is a statistical graph showing the effects of compounds 1a-1j in Examples 1-10 and compound 9 in Comparative Example 1 on HL-60 cell colony formation.

[0035] Figure 2 is a statistical chart showing the effects of compounds 1a-1j in Examples 1-10 and compound 9 in Comparative Example 1 on the formation of Kasumi-1 cell clones.

[0036] Figure 3 is a statistical graph showing the effect of compound 1a on iron ion concentration in HL-60 cells in Example 1.

[0037] Figure 4 is a statistical graph showing the effect of compound 1a on iron ion concentration in Kasumi-1 cells in Example 1.

[0038] Figure 5 shows the results of the acute toxicity test of compound 1a in Example 1. Detailed Implementation

[0039] To enable those skilled in the art to better understand the present invention, the technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort should fall within the scope of protection of the present invention.

[0040] Example

[0041] The present invention will be further illustrated below with reference to specific embodiments and comparative embodiments. The following specific embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the following embodiments, and are not in particular limited to the types of raw materials used in the following specific embodiments.

[0042] I. The sources of raw materials for the examples and comparative examples are as follows:

[0043] Unless otherwise specified, all raw materials used in the embodiments of this invention are commercially available.

[0044] The method for preparing tetrahydroisoquinoline compounds according to embodiments of the present invention includes the following steps:

[0045] Compound 2 (1.0 mmol), TBTU (1.0 mmol), and K2CO3 (1.5 mmol) were added sequentially to a round-bottom flask. Dry N,N-dimethylformamide (10 mL) was added, and the mixture was stirred at room temperature for 1 hour. Then, 6-amino-1,2,3,4-tetrahydroisoquinoline-2-carboxylic acid-2-methylpropyl-2-yl ester (1.0 mmol) was added, and the mixture was magnetically stirred at 60 °C for 12 hours. The mixture was then extracted with ethyl acetate (3 × 150 mL). The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The solid obtained by rotary evaporation was purified by column chromatography to give a yellow solid, which yielded compound 4 (yield 61%–83%).

[0046] Compound 4 (1.0 mmol) was added to 5 mL of methanol, followed by 1 mL of TFA. After stirring at room temperature for 3 hours, saturated sodium carbonate solution was added to adjust the pH to about 8. The mixture was then extracted with ethyl acetate (3 × 150 mL). The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain crude compound 5. Compound 6 (1.0 mmol) and DBU (1.2 mmol) were then added and dissolved in N,N-dimethylformamide (10 mL). The mixture was magnetically stirred at 60 °C for 8 hours. The mixture was then extracted with ethyl acetate (3 × 150 mL). The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The solid obtained by rotary evaporation was purified by column chromatography to obtain a yellow solid, which yielded compound 7 (yield 59%–77%).

[0047] Compound 7 (1.0 mmol), compound 8 (1.0 mmol), cesium carbonate (1.2 mmol), and Pd(OAc)2 (1.0 mmol) were added to 10 mL of methanol and stirred at 60 °C for 8 h. After the reaction was completed, the solid was removed by filtration. The solid was washed with methanol (3 × 10 mL), and the filtrate was evaporated to dryness. The solid was purified by column chromatography to obtain a yellow solid, which was compound 1.

[0048] Example 1

[0049] Compound 2 used in Example 1 was alanine, and compound 1a was prepared as follows: The structural characterization data are as follows:

[0050] 1 H NMR (400MHz, CD3OD) δ7.65(t,J=1.8Hz,1H),7.42(d,J=8.8Hz,1H),7.37(m,1H),7.31 (t,J=7.9Hz,1H),7.15(dd,J=9.4,5.6Hz,1H),7.07-7.03(m,2H),6.98(dd,J=8.8,2. 2Hz,1H),6.87-6.80(m,2H),4.39(s,2H),3.82(q,J=6.0Hz,1H),3.57(t,J=6.0Hz,2H ),2.94(m,6H),2.85(s,4H),2.48(s,3H),1.29(d,J=6.0Hz,3H).ESI-MS:508.18[MH] - .

[0051] Example 2

[0052] Compound 2 used in Example 2 was aspartic acid, and compound 1b was prepared as follows: The structural characterization data are as follows:

[0053] 1 H NMR (400MHz, CD3OD) δ14.l(bs,1H),7.66(t,J=1.8Hz,1H),7.45(d,J=8.8Hz,1H),7. 34(m,1H),7.33(t,J=7.9Hz,1H),7.17(dd,J=9.4,5.6Hz,1H),7.07-7.03(m,2H),6.9 8(dd,J=8.8,2.2Hz,1H),6.87-6.80(m,2H),4.39(s,2H),3.84(t,J=6.0Hz,1H),3.5 8(t,J=6.0Hz,2H),2.94(m,6H),2.85-2.60(m,6H),2.48(s,3H).ESI-MS:552.17[MH] - .

[0054] Example 3

[0055] Compound 2 used in Example 3 was glutamic acid, and the resulting compound 1c was: The structural characterization data are as follows:

[0056] 1 H NMR (400MHz, CD3OD) δ12.88(bs,1H),7.68(t,J=1.8Hz,1H),7.43(d,J=8.8Hz,1H ),7.36(m,1H),7.31(t,J=7.9Hz,1H),7.15(dd,J=9.4,5.6Hz,1H),7.07-7.03(m, 2H),6.98(dd,J=8.8,2.2Hz,1H),6.87-6.80(m,2H),4.33(s,2H),3.57(t,J=6.0Hz,2H),3.41( t,J=6.0Hz,1H),2.94(m,6H),2.84(s,4H),2.47(s,3H),2.18-2.07(m,4H).ESI-MS:566.21[MH] - .

[0057] Example 4

[0058] Compound 2 used in Example 4 was acetic acid, and the resulting compound 1d was: The structural characterization data are as follows:

[0059] 1H NMR (400MHz, CD3OD) δ7.65(t,J=1.8Hz,1H),7.42(d,J=8.8Hz,1H),7.35(m,1 H),7.32(t,J=7.9Hz,1H),7.13(dd,J=9.4,5.6Hz,1H),7.07-7.03(m,2H),6. 98(dd,J=8.8,2.2Hz,1H),6.87-6.80(m,2H),4.39(s,2H),3.58(t,J=6.0Hz, 2H),2.94(m,6H),2.84(s,4H),2.46(s,3H),2.05(s,3H).ESI-MS:479.25[MH] - .

[0060] Example 5

[0061] Compound 2 used in Example 5 was ethyl bicarbonate, and the resulting compound 1e was: The structural characterization data are as follows:

[0062] 1 H NMR (400MHz, CD3OD) δ7.65(t,J=1.8Hz,1H),7.43(d,J=8.8Hz,1H),7.37(m,1H),7.31 (t,J=7.9Hz,1H),7.17(dd,J=9.4,5.6Hz,1H),7.07-7.03(m,2H),6.98(dd,J=8.8,2. 2Hz,1H),6.87-6.80(m,2H),4.40(s,2H),4.12(q,J=6.0Hz,2H),3.57(t,J=6.0Hz,2H ),2.94(m,6H),2.82(s,4H),2.48(s,3H),1.26(t,J=6.0Hz,3H).ESI-MS:509.18[MH] - .

[0063] Example 6

[0064] Compound 2 used in Example 6 was glutamic acid, and the resulting compound 1f was: The structural characterization data are as follows:

[0065] 1H NMR(400MHz,CD3OD)δ7.60(t,J=1.8Hz,1H),7.36(d,J=8.8Hz,1H),7.32(m,1H), 7.28(t,J=7.9Hz,1H),7.15(dd,J=9.4,5.6Hz,1H),7.06-7.03(m,2H),6.98(dd, J=8.8,2.2Hz,1H),6.87-6.80(m,2H),4.39(s,2H),3.82(q,J=6.0Hz,1H),3.57(t,J=6.0H z,2H),2.95(m,6H),2.85(s,4H),2.47(s,3H),1.28(d,J=6.0Hz,3H).ESI-MS:508.18[MH] - .

[0066] Example 7

[0067] Compound 2 used in Example 7 was L-alanine, compound 3 was XX, and 1g of the obtained compound was: The structural characterization data are as follows:

[0068] 1 H NMR (400MHz, CD3OD) δ7.65(t,J=1.8Hz,1H),7.40(d,J=8.8Hz,1H),7.35(m,1H),7.26 (t,J=7.9Hz,1H),7.15(dd,J=9.4,5.6Hz,1H),7.07-7.04(m,2H),7.00(dd,J=8.8,2. 2Hz,1H),6.88-6.82(m,2H),4.39(s,2H),3.82(q,J=6.0Hz,1H),3.57(t,J=6.0Hz,2H ),2.95(m,6H),2.85(s,4H),2.47(s,3H),1.31(d,J=6.0Hz,3H).ESI-MS:508.17[MH] - .

[0069] Example 8

[0070] Compound 2 used in Example 8 was L-alanine, compound 3 was XX, and the resulting compound 1h was: The structural characterization data are as follows:

[0071] 1H NMR (400MHz, CD3OD) δ7.66 (t, J=1.8Hz, 1H), 7.41 (d, J=8.8Hz, 1H), 7.35 (m, 1H), 7.26 (t,J=7.9Hz,1H),7.16(dd,J=9.4,5.6Hz,1H),7.07-7.04(m,2H),7.00(dd,J=8.8,2. 2Hz,1H),6.87-6.81(m,2H),4.40(s,2H),3.83(q,J=6.0Hz,1H),3.57(t,J=6.0Hz,2H ),2.95(m,6H),2.85(s,4H),2.47(s,3H),1.28(d,J=6.0Hz,3H).ESI-MS:508.20[MH] - .

[0072] Example 9

[0073] Compound 2 used in Example 9 was glutamine, and the prepared compound 1i was: The structural characterization data are as follows:

[0074] 1 H NMR (400MHz, CD3OD) δ7.66(t,J=1.8Hz,1H),7.42(d,J=8.8Hz,1H),7.38(m,1H),7.3 3(t,J=7.9Hz,1H),7.17(dd,J=9.4,5.6Hz,1H),7.09-7.03(m,2H),6.98(dd,J=8.8, 2.2Hz,1H),6.87-6.80(m,2H),4.39(s,2H),3.55(t,J=6.0Hz,2H),3.35(t,J=6.0Hz ,1H),2.94(m,6H),2.85(s,4H),2.48(s,3H),2.15-2.08(m,4H).ESI-MS:551.30[MH] - .

[0075] Example 10

[0076] Compound 2 used in Example 10 was asparagine, and the obtained compound 1j was: The structural characterization data are as follows:

[0077] 1H NMR (400MHz, CD3OD) δ7.66(t,J=1.8Hz,1H),7.43(d,J=8.8Hz,1H),7.37(m,1H),7.33(t,J =7.9Hz,1H),7.16(dd,J=9.4,5.6Hz,1H),7.08-7.03(m,2H),6.98(dd,J=8.8,2.2Hz,1H),6 .88-6.80(m,2H),4.39(s,2H),4.01(t,J=6.0Hz,1H),3.55(t,J=6.0Hz,2H),2.94(m,6H),2 .90(d,J=6.0Hz,1H),2.85(s,4H),2.62(d,J=6.0Hz,1H),2.48(s,3H).ESI-MS:551.30[MH] - .

[0078] Comparative Example 1

[0079] The compound provided in Comparative Example 1 does not possess [the following characteristics]. Compound 9, with the following structural formula:

[0080] Example 11 Preparation of Injection Solution

[0081] Compound 1a prepared in Example 1 was dissolved in a small amount of DMSO, and then water for injection was added according to standard procedures. After fine filtration, the solution was filled, sealed, and sterilized to prepare an injection solution.

[0082] Example 12 Preparation of tablets

[0083] Compound 1a prepared in Example 1 was granulated and compressed into tablets with cyclodextrin at a weight ratio of 6:1 to obtain tablets.

[0084] Example 13: Preparation of Capsules

[0085] Compound 1a prepared in Example 1 was mixed with microcrystalline cellulose in a weight ratio of 6:1 to form capsules.

[0086] Example 14: Testing the inhibitory effect of compound 1a-1j on leukemia cells

[0087] 1. Experimental Methods

[0088] Cell source: The cell lines used in this study are human leukemia cells HL-60 and Kasumi-1, derived from ATCC.

[0089] Cell culture: Cells were cultured at 37°C in a 5% CO2 incubator. After being removed from the cryovial, cells were thawed as quickly as possible in a 37°C water bath and immediately placed in 20% fetal bovine serum-modified IMDM medium, with 10 μg / mL of DNASE I added. Cells were centrifuged at 1500 rpm for 5 minutes and then resuspended in complete culture medium at a concentration of 2-5 × 10⁻⁵. 6 Cells / mL; Thawed cells were then cultured in complete medium supplemented with 10% fetal bovine serum (FBS) and BIT (bovine serum albumin 4 g / L, insulin 5 μg / mL, transferrin 60 μg / mL, all from Sigma-Aldrich). To promote cell growth and maintenance, specific cytokines and components were added: 50 ng / mL FLT3 ligand, 10 ng / mL IL-6, 50 ng / mL stem cell factor (SCF), 25 ng / mL thrombopoietin (TPO), 10 ng / mL IL-3, and 10 ng / mL granulocyte colony-forming factor (G-CSF). In addition, the medium also contained 50 μM β-mercaptoethanol (Sigma-Aldrich).

[0090] Test method: Colony formation was detected using the L-CFU method, with cells at a concentration of 1×10⁻⁶. 5 L-CFU (>10 ng / mL) were seeded in H4230 medium supplemented with 10% IMDM, 50 ng / mL FLT3 ligand, 10 ng / mL IL-6, 50 ng / mL stem cell factor (SCF), 25 ng / mL thrombopoietin (TPO), 10 ng / mL IL-3, and 10 ng / mL granulocyte colony factor (G-CSF). The patients were divided into control and treatment groups, with an administration concentration of 1 μM. On day 7, L-CFU (>10 cell colonies) were counted using an inverted microscope.

[0091] 2. Experimental Results

[0092] L-CFU analysis showed that compounds 1a-1j effectively reduced leukemia cell colony formation (Figures 1 and 2), and the compounds of this invention exhibited stronger anticancer effects compared to compound 9. This also demonstrates that the introduction of the amide group enhanced the cell membrane penetration ability and improved the activity of the compounds, especially compounds 1a, 1d, 1e, and 1f, which showed extremely significant effects in reducing leukemia cell colony formation, with 1a exhibiting the best overall effect.

[0093] Example 15: Testing the effect of compound 1a on intracellular iron ion levels

[0094] 1. Experimental Methods

[0095] Intracellular Fe2+ Level assay: HL-60 and Kasumi-1 cells were seeded in 96-well laser confocal microscopy plates, with 5000 cells per well in 100 μL of medium. After overnight incubation, the old medium was discarded, and the cells were treated with medium containing 0.5 μM compound 1a for 6 hours. A probe working solution containing 1 μM FerroOrange and 1 μg / mL Hoechst 33342 was prepared using serum-free medium and added to the 96-well plates. The cells were incubated at 37°C for 30 min, followed by washing with 1×PBS once. Cell analysis was performed using an FV3000 laser confocal microscope or a Cellmics ArrayScan Vti high-content system.

[0096] 2. Experimental Results

[0097] The results showed that compound 1a could significantly reduce the concentration of iron ions in leukemia cells (Figures 3 and 4).

[0098] Example 16 Acute toxicity test of compound 1a

[0099] 1. Experimental Methods

[0100] The experiment used SPF-grade C57BL / 6 mice (6-7 weeks old) which were housed in an SPF-grade animal room at a temperature of 21-24℃ and a humidity of 50-70%. They were fed SPF-grade mouse food and given sterile water. After one week of housekeeping in the animal room, no abnormalities were observed. The mice were then randomly divided into 4 groups (control group, 100mg / kg, 200mg / kg, and 400mg / kg), with 6 mice in each group. Half of the mice were male and half were kept in separate cages.

[0101] The mice were fasted for 12 hours before administration, but allowed free access to water. The mice were then administered the drug by gavage, and were fed normally 4 hours after administration. Within one week of administration, the mice were weighed on days 1, 3, 5, and 7, and any abnormal behavior or death was observed.

[0102] 2. Experimental Results

[0103] As shown in Figure 5, compared with the control group, the mice in the drug-treated group did not show significant changes in body weight, nor did they exhibit acute toxicity symptoms such as death, abnormal behavior, or decreased appetite.

[0104] Based on the test data on the inhibitory effect of tetrahydroisoquinoline compounds on leukemia cells, their influence on intracellular iron ion levels, and acute toxicity, it is sufficient to demonstrate that the tetrahydroisoquinoline compounds prepared by Examples 1-10 have a significant inhibitory effect on leukemia cells and low toxicity to organisms. This not only expands the types of drugs available for clinical treatment of leukemia but is also suitable for large-scale production.

[0105] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. The application of a tetrahydroisoquinoline compound in the preparation of an anti-leukemia drug, characterized in that, The tetrahydroisoquinoline compounds have the structure of formula (I): in, It is a substituted amide group formed by the carboxylic acid and amino group on an amino acid, or R1 is hydrogen or C. 1～12 Alkyl, C 1～12 Alkoxy, C 1～12 Alkyl group, C 1～12 Any substituted amide group among alkyl ester groups; R2 is X is any one of C or N atoms, R 21 for R 211 It may be mono- or poly-substituted, selected from one or more of hydrogen, halogen, unsubstituted or substituted piperazine, indole, and morpholine, wherein the substituent of the substituted piperazine is C. 1～6 Alkyl, halogen, C 1～6 Any one of alkoxy, hydroxy, or amino groups.

2. The application according to claim 1, characterized in that, The The substituted amide group is formed by the carboxylic acid and amino group of any one of the amino acids selected from alanine, phenylalanine, L-alanine, cysteine, selenocysteine, aspartic acid, asparagine, glutamic acid, glutamine, glycine, histidine, leucine, isoleucine, lysine, pyrrolidone, methionine, proline, arginine, serine, threonine, valine, tryptophan, and tyrosine, or R1 is hydrogen or carbon. 1～12 Alkyl, C 1～12 Any substituted amide group among alkoxy groups; The R 211 The substituted piperazine has a C substituent. 1～3 alkyl.

3. The application according to claim 2, characterized in that, The The substituted amide group is formed by the carboxylic acid and amino group of any one of the amino acids selected from alanine, L-alanine, aspartic acid, asparagine, glutamic acid, and glutamine, or R1 is C. 1～12 Alkyl, C 1～12 Any substituted amide group of alkoxy groups.

4. The application according to claim 1, characterized in that, The tetrahydroisoquinoline compounds have any one of the following structural formulas:

5. The application according to claim 1, characterized in that, The tetrahydroisoquinoline compounds may be used in any of their pharmaceutically acceptable salts, solvates, or chiral isomers.

6. The application according to claim 5, characterized in that, The pharmaceutically acceptable salt is obtained by reacting tetrahydroisoquinoline compounds with inorganic or organic acids.

7. The application according to claim 6, characterized in that, The organic acid is any one of citric acid, acetic acid, oxalic acid, isonicotinic acid, lactic acid, salicylic acid, acidic citric acid, tartaric acid, oleic acid, tannic acid, pantothenic acid, tartaric acid, ascorbic acid, succinic acid, maleic acid, gentian acid, fumaric acid, gluconic acid, glucuronic acid, succinic acid, formic acid, benzoic acid, glutamic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, or dihydroxynaphthyl acid.

8. The application according to claim 6, characterized in that, The inorganic acid is any one of hydrochloric acid, phosphoric acid, sulfuric acid, or nitric acid.

9. The application according to claim 1, characterized in that, The drug is any one of oral, injectable, topical, or inhaled formulations.

10. The application according to claim 1, characterized in that, The oral medication is any one of capsules, tablets, pills, or granules; the inhalant is a spray.

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