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

Abstract

The present invention discloses the use of a tetrahydroisoquinoline compound in the preparation of an anti-leukemia drug. The compound has a significant inhibitory effect on leukemia cells and low toxicity to organisms, expanding the range of clinical leukemia treatment drugs and being suitable for large-scale production.

Description

Technical Field

[0001] The present invention relates to the technical field of biomedicine, and in particular to the application of a tetrahydroisoquinoline compound in the preparation of an anti-leukemia drug. Background Art

[0002] Malignant tumors, a major health challenge that threatens human life, continue to rank first among the causes of death worldwide.

[0003] Although advances in technology and medical technology have led to some progress in cancer treatment, and the survival rates of patients with common malignant tumors have improved, malignant tumors remain a significant threat to human health. Hematologic malignancies, as a special type of malignancy, have attracted particular attention due to their wide age range of onset and difficulty in treatment.

[0004] Hematologic malignancies, as the name suggests, are a class of diseases caused by the malignant transformation and proliferation of cells in the hematopoietic system. These cancers can occur in a wide range of ages, from infants to the elderly, making them one of the cancers with the widest age range of onset. Hematologic malignancies are primarily characterized by abnormalities in the quantity and quality of white blood cells and their immature cells (precursor cells or leukemic 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 complex, encompassing numerous different 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 their diagnosis and treatment more difficult. For example, acute myeloid leukemia (AML) is a clonal hematologic malignancy caused by abnormal development of hematopoietic stem or progenitor cells, and its incidence increases with age. Acute lymphoblastic leukemia (ALL) is caused by an unexplained maturation disorder of lymphoblasts, which leads to excessive proliferation, suppressing normal bone marrow hematopoietic tissue and causing 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 is a combination of hematopoietic stem cell transplantation and chemotherapy. However, while these approaches can improve patient survival to a certain extent, many patients still experience relapse and no response to treatment. Furthermore, the current range of drugs for the treatment of leukemia remains limited, failing to meet the growing clinical needs. Therefore, there is an urgent need to develop more anti-leukemia drugs, especially those with more effective targeting and enhanced cancer cell membrane penetration, to provide more options for medical research and clinical application. Summary of the Invention

[0007] In view of the defects in the prior art, the present invention proposes the use of a tetrahydroisoquinoline compound in the preparation of an anti-leukemia drug.

[0008] The present invention provides an application of a tetrahydroisoquinoline compound in the preparation of an anti-leukemia drug. The tetrahydroisoquinoline compound has a structure of formula (I):

[0009]

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

[0011] R2 is X is any one of C or N atoms, R 21 for R 211 is monosubstituted or polysubstituted and is 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 of alkoxy, hydroxy or amino;

[0012] On the one hand, the tetrahydroisoquinoline compounds of the present invention can target and inhibit the interaction between nuclear receptor coactivator 4 and ferritin heavy chain, inhibit iron autophagy, and thus inhibit the proliferation of tumor cells, especially the inhibition effect on leukemia cells is significant, achieving the effect of treating leukemia; on the other hand, the inventors found that the amide group in the tetrahydroisoquinoline compounds This can improve the drug's targeting and ability to penetrate cancer cell membranes, thereby enhancing the compound's activity. Since cancer cell surfaces typically carry a weak negative charge, positively charged amide molecules are more easily absorbed by the cancer cell surface, resulting in a high anticancer effect. Therefore, the tetrahydroisoquinoline compounds of the present invention offer a wider range of options for medical research and clinical applications.

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

[0014] The R 211 is a substituted piperazine, wherein the substituent of the substituted piperazine is C 1～3 alkyl.

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

[0016] The following is an exemplary synthesis route of a tetrahydroisoquinoline compound, wherein R2 of the exemplary compound 1 is X is a C atom, R 21 for R 211 It is a monosubstituted piperazine, where the substituent of the substituted piperazine is methyl:

[0017]

[0018] The following is a method for preparing compound 1, comprising the following steps:

[0019] Compound 2 (0.9-1.1 mmol), TBTU (0.9-1.1 mmol) and K2CO3 (1.4-1.6 mmol) were added to a round-bottom flask in sequence, followed by the addition of dry N,N-dimethylformamide (9-11 mL). The mixture was stirred at room temperature for 50-70 min, followed by the addition of compound 3 (0.8-1.2 mmol). The mixture was stirred at 60°C for 11-13 hours under magnetic stirring, and then extracted with ethyl acetate (3 × 150 mL, i.e., three times with 150 mL of ethyl acetate). The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The solid obtained by spin drying was separated and purified by column chromatography to obtain a yellow solid to obtain compound 4 (yield 61% to 83%).

[0020] 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, a saturated sodium carbonate solution was added to adjust the pH to about 8, and then extracted with ethyl acetate (3×150 mL). The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a 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 stirred under magnetic stirring at 60°C for 7-9 hours, and 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 spin drying was separated and purified by column chromatography to obtain a yellow solid to obtain compound 7 (yield 59% to 77%).

[0021] 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 filtered out and rinsed with methanol (3×10 mL, i.e., rinsed three times with 10 mL of methanol). The filtrate was then dried and purified by column chromatography to obtain a yellow solid to obtain compound 1.

[0022] Furthermore, the tetrahydroisoquinoline compound has any one of the following structural formulas:

[0023]

[0024] Preferably

[0025]

[0026] Furthermore, the tetrahydroisoquinoline compound can be used in the form of any one of its pharmaceutically acceptable salts, solvates or chiral isomers.

[0027] Furthermore, the pharmaceutically acceptable salt is obtained by reacting a tetrahydroisoquinoline compound with an inorganic acid or an organic acid.

[0028] Furthermore, the organic acid is any one of citric acid, acetic acid, oxalic acid, isonicotinic acid, lactic acid, salicylic acid, acid citric acid, tartaric acid, oleic acid, tannic acid, pantothenic acid, tartaric acid, ascorbic acid, succinic acid, maleic acid, gentisic acid, fumaric acid, gluconic acid, glucuronic acid, sugar acid, formic acid, benzoic acid, glutamic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid or pamoic acid.

[0029] Furthermore, the inorganic acid is any one of hydrochloric acid, phosphoric acid, sulfuric acid or nitric acid.

[0030] Furthermore, the medicine is any one of an oral preparation, an injection, an external preparation or an inhalation preparation.

[0031] Furthermore, the oral dosage form is any one of capsules, tablets, pills, and granules; and the inhalant is a spray.

[0032] In summary, compared with the prior art, the present invention achieves the following technical effects:

[0033] (1) The tetrahydroisoquinoline compounds provided by the present invention can significantly inhibit the proliferation of leukemia cells;

[0034] (2) The tetrahydroisoquinoline compounds provided by the present invention have high targeting and stronger ability to penetrate cancer cell membranes, which can enhance the activity of the compounds and have a high anti-cancer effect;

[0035] (3) The preparation process of the tetrahydroisoquinoline compounds provided by the present invention is simple and easy to produce on a large scale. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the following briefly introduces the drawings required for use in the embodiments. It should be understood that the following drawings only illustrate certain embodiments of the present invention and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other relevant drawings can be obtained based on these drawings without paying any creative work.

[0037] Figure 1 This 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 clone formation.

[0038] Figure 2 This is a statistical graph showing the effects of compounds 1a-1j in Examples 1-10 and compound 9 in Comparative Example 1 on the colony formation of Kasumi-1 cells.

[0039] Figure 3 This is a statistical graph showing the effect of compound 1a in Example 1 on the iron ion concentration in HL-60 cells.

[0040] Figure 4 This is a statistical graph showing the effect of compound 1a in Example 1 on the iron ion concentration in Kasumi-1 cells.

[0041] Figure 5 These are the results of the acute toxicity test of compound 1a in Example 1. DETAILED DESCRIPTION

[0042] In order to help those skilled in the art better understand the solutions of the present invention, the technical solutions in the embodiments of the present invention are clearly and completely described below. Obviously, the embodiments described are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without making any creative efforts should fall within the scope of protection of the present invention.

[0043] Example

[0044] The present invention is further described below with reference to specific examples and comparative examples. The following specific examples are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the following examples, and are particularly not limited to the types of the various component raw materials used in the following specific examples.

[0045] 1. The sources of raw materials for the embodiments and comparative examples are as follows:

[0046] Unless otherwise specified, the raw materials used in the examples of the present invention are commercially available.

[0047] The method for preparing the tetrahydroisoquinoline compound according to the embodiment of the present invention comprises the following steps:

[0048] Compound 2 (1.0 mmol), TBTU (1.0 mmol) and K2CO3 (1.5 mmol) were added to a round-bottom flask in sequence, and dry N,N-dimethylformamide (10 mL) was added. The mixture was stirred at room temperature for 1 hour, and then 6-amino-1,2,3,4-tetrahydroisoquinoline-2-carboxylic acid-2-methylprop-2-yl ester (1.0 mmol) was added. The mixture was stirred at 60°C for 12 hours, and 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 spin drying was separated and purified by column chromatography to obtain a yellow solid to obtain compound 4 (yield 61% to 83%).

[0049] 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, a saturated sodium carbonate solution was added to adjust the pH to about 8, and then extracted with ethyl acetate (3×150 mL). The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a 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 stirred under magnetic stirring at 60°C for 8 hours, and 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 spin drying was separated and purified by column chromatography to obtain a yellow solid to obtain compound 7 (yield 59% to 77%).

[0050] 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 filtered out and rinsed with methanol (3×10 mL). The filtrate was then dried and purified by column chromatography to obtain a yellow solid to obtain compound 1.

[0051] Example 1

[0052] The compound 2 used in Example 1 is alanine, and the obtained compound 1a is: The structural characterization data are as follows:

[0053] 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] - .

[0054] Example 2

[0055] The compound 2 used in Example 2 is aspartic acid, and the obtained compound 1b is: The structural characterization data are as follows:

[0056] 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] - .

[0057] Example 3

[0058] The compound 2 used in Example 3 is glutamic acid, and the obtained compound 1c is: The structural characterization data are as follows:

[0059] 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] - .

[0060] Example 4

[0061] The compound 2 used in Example 4 is acetic acid, and the obtained compound 1d is: The structural characterization data are as follows:

[0062] 1 H 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] - .

[0063] Example 5

[0064] The compound 2 used in Example 5 is ethyl bicarbonate, and the obtained compound 1e is: The structural characterization data are as follows:

[0065] 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] - .

[0066] Example 6

[0067] The compound 2 used in Example 6 is glutamic acid, and the obtained compound 1f is: The structural characterization data are as follows:

[0068] 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=6.0Hz,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] - .

[0069] Example 7

[0070] The compound 2 used in Example 7 is L-alanine, the compound 3 is XX, and the obtained compound 1g is: The structural characterization data are as follows:

[0071] 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] - .

[0072] Example 8

[0073] The compound 2 used in Example 8 is L-alanine, the compound 3 is XX, and the obtained compound 1h is: The structural characterization data are as follows:

[0074] 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] - .

[0075] Example 9

[0076] The compound 2 used in Example 9 is glutamine, and the obtained compound 1i is: The structural characterization data are as follows:

[0077] 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] - .

[0078] Example 10

[0079] The compound 2 used in Example 10 is asparagine, and the obtained compound 1j is: The structural characterization data are as follows:

[0080] 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] - .

[0081] Comparative Example 1

[0082] The compound provided in Comparative Example 1 is not The compound 9 of the group has the following structural formula:

[0083]

[0084] Example 11 Preparation of injection

[0085] The compound 1a prepared in Example 1 was dissolved in a small amount of DMSO, and then added with water for injection according to conventional methods. The mixture was finely filtered, sealed and sterilized to prepare an injection solution.

[0086] Example 12 Preparation of tablets

[0087] The compound 1a prepared in Example 1 and cyclodextrin were granulated and tableted at a weight ratio of 6:1 to obtain tablets.

[0088] Example 13 Preparation of capsules

[0089] The compound 1a prepared in Example 1 and microcrystalline cellulose were prepared into capsules at a weight ratio of 6:1.

[0090] Example 14 Testing the inhibitory effect of compounds 1a-1j on leukemia cells

[0091] 1. Experimental methods

[0092] Cell source: The cell lines used in the present invention are human leukemia cells HL-60 and Kasumi-1, which are obtained from ATCC.

[0093] Cell culture: Cells were cultured in a 37°C, 5% CO2 incubator. After removal from the cryostat, cells were thawed as quickly as possible in a 37°C water bath and immediately placed in IMDM medium supplemented with 20% fetal bovine serum and 10 μg / mL DNASE I. Cells were centrifuged at 1500 rpm for 5 minutes and then resuspended in complete culture medium at a concentration of 2-5 × 10 6 cells / mL; the thawed cells were then cultured in complete medium. IMDM medium was 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, some 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 factor (G-CSF). In addition, the culture medium also contained 50 μM β-mercaptoethanol (Sigma-Aldrich).

[0094] Test method: L-CFU method was used to detect clone formation, and the cells were counted as 1×10 5 / mL was inoculated in H4230 culture medium supplemented with 10% IMDM, 50ng / mL FLT3 ligand, 10ng / mL IL-6, 50ng / mL stem cell factor (SCF), 25ng / mL thrombopoietin (TPO), 10ng / mL IL-3 and 10ng / mL granulocyte colony factor (G-CSF). The cells were divided into a control group and a drug-treated group. The drug-treated group was 1μM. On day 7, L-CFU (>10 cell colonies) were counted using an inverted microscope.

[0095] 2. Experimental results

[0096] L-CFU analysis showed that compounds 1a-1j can effectively reduce the formation of leukemia cell clones ( Figure 1 and Figure 2 ), and the compounds of the present invention have stronger anticancer effects than compound 9. This also proves that the introduction of the amide group enhances the ability to penetrate the cell membrane and improves the activity of the compounds. In particular, 1a, 1d, 1e, and 1f have extremely significant effects in reducing the formation of leukemia cell colonies, among which 1a has the best overall effect.

[0097] Example 15: Effect of Compound 1a on Intracellular Iron Levels

[0098] 1. Experimental methods

[0099] Intracellular Fe 2+ Level determination: HL-60 and Kasumi-1 cells were seeded in 96-well laser confocal microplates, with 5000 cells per well in 100 μL of culture medium. After the cells were cultured overnight in the culture medium, the old culture medium was discarded and the cells were treated with culture medium containing 0.5 μM compound 1a for 6 hours. The probe working solution containing 1 μM FerroOrange and 1 μg / mL Hoechst 33342 was prepared in serum-free culture medium and added to the 96-well plate. The cells were incubated in a 37°C incubator for 30 minutes, and then the cells were washed once with 1× PBS. Cell detection was performed using an FV3000 laser confocal microscope or a Cellomics ArrayScan Vti high-content system.

[0100] 2. Experimental results

[0101] The results showed that compound 1a could significantly reduce the concentration of iron ions in leukemia cells ( Figure 3 and Figure 4 ).

[0102] Example 16 Acute toxicity test of compound 1a

[0103] 1. Experimental methods

[0104] The experiment used SPF-grade C57BL / 6 mice (6-7 weeks old), which were raised in an SPF animal room at a temperature of 21-24°C and a humidity of 50-70%. They were fed SPF-grade mouse feed and sterile drinking water. After one week of breeding in the animal room after purchase, no abnormalities were observed. They were then randomly divided into four groups (control group, 100 mg / kg, 200 mg / kg, and 400 mg / kg), with 6 mice in each group, half male and half female, and housed in separate cages at the same time.

[0105] The mice were fasted for 12 hours before administration and allowed to drink water freely. The mice were then gavage-administered and fed normally 4 hours after administration. Within one week of administration, the mice were weighed on the 1st, 3rd, 5th and 7th days, and their abnormal behavior and death were observed.

[0106] 2. Experimental results

[0107] like Figure 5 As shown, compared with the control group, the mice in the drug-treated group did not show obvious weight changes, nor did they experience acute toxic phenomena such as mouse death, abnormal behavior, and decreased appetite.

[0108] Based on the test data on the inhibitory effect of tetrahydroisoquinoline compounds on leukemia cells, the effect on intracellular iron ion levels, and acute toxicity, it is sufficient to show that the tetrahydroisoquinoline compounds prepared by Examples 1-10 have significant inhibitory effects on leukemia cells and low toxicity to organisms, which not only expands the types of drugs for the clinical treatment of leukemia, but also is suitable for large-scale production.

[0109] 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 in the scope of protection of the present invention.

Claims

1. Use of a tetrahydroisoquinoline compound in the preparation of an anti-leukemia drug, characterized in that: The tetrahydroisoquinoline compound has the structure of formula (I): in, It is a substituted amide group formed by the carboxylic acid and amino group on any one of the amino acids of alanine, glutamic acid, L-alanine, glutamine, and asparagine, or R1 is any one of methyl and ethoxy; R2 is X is any one of C or N atoms, R 21 for R 211 It is a methyl-substituted piperazine.

2. The use according to claim 1, characterized in that The tetrahydroisoquinoline compound has any one of the following structural formulas:

3. The use according to claim 1, characterized in that The tetrahydroisoquinoline compound can be used in the form of a pharmaceutically acceptable salt.

4. The use according to claim 3, characterized in that The pharmaceutically acceptable salt is obtained by reacting a tetrahydroisoquinoline compound with an inorganic acid or an organic acid.

5. The use according to claim 4, characterized in that The organic acid is any one of citric acid, acetic acid, oxalic acid, isonicotinic acid, lactic acid, salicylic acid, acid citric acid, oleic acid, tannic acid, pantothenic acid, tartaric acid, ascorbic acid, succinic acid, maleic acid, gentisic acid, fumaric acid, gluconic acid, glucuronic acid, sugar acid, formic acid, benzoic acid, glutamic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid or pamoic acid.

6. The use according to claim 4, characterized in that The inorganic acid is any one of hydrochloric acid, phosphoric acid, sulfuric acid or nitric acid.

7. The use according to claim 1, characterized in that The medicine is any one of an oral preparation, an injection, an external preparation or an inhalation preparation.

8. The use according to claim 7, characterized in that The oral preparation is any one of capsules, tablets, pills and granules; the inhalant is a spray.

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