Application of tetrahydroisoquinoline compound in preparation of anti-leukemia drugs

Abstract

The invention discloses an application of a tetrahydroisoquinoline compound in preparation of an anti-leukemia drug. The compound has a remarkable inhibition effect on leukemia cells, has low toxicity to organisms, not only expands the variety of drugs for clinical treatment of leukemia, but also is suitable for large-scale production.

Description

Technical Field

[0001] The present invention relates to the technical field of biomedicine, and in particular to 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, continues to rank first among the causes of death worldwide.

[0003] Although we have made some progress in cancer treatment with the advancement of science and technology and the improvement of medical standards, and the survival rate of patients with common types of malignant tumors has increased, malignant tumors are still a huge threat to human health. Among them, blood tumors, as a special type of malignant tumor, have attracted much attention due to their wide age range of onset and difficulty in treatment.

[0004] Hematological tumors, as the name suggests, are a type of disease caused by the cancerous transformation and malignant proliferation of hematopoietic system cells. This type of cancer has a very wide age range of onset, ranging from infants to the elderly, making it one of the cancers with the widest age range of onset. The main feature of hematological tumors 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 inhibit the normal hematopoietic function of the bone marrow and infiltrate into tissues and organs such as the liver, spleen, and lymph nodes, causing patients to experience symptoms such as bleeding, bruising, fatigue, and increased risk of infection.

[0005] The classification of blood tumors is very complex, including many different types of leukemia, lymphoma, etc. These diseases differ in pathogenesis, clinical manifestations and treatment methods. Moreover, most blood tumors are accompanied by genetic changes, which makes the diagnosis and treatment of blood tumors more difficult. For example, acute myeloid leukemia is a clonal blood tumor caused by abnormal development of hematopoietic stem cells or progenitor cells, and its incidence increases with age. Acute lymphocytic leukemia is caused by excessive proliferation of lymphoblasts due to unexplained maturation disorders, which in turn suppresses normal bone marrow hematopoietic tissue and causes infiltration of peripheral blood vessels, bone marrow, systemic lymph nodes, spleen and liver. Acute lymphocytic leukemia is particularly common in adults.

[0006] At present, the main treatment for leukemia is the combination of hematopoietic stem cell transplantation and drug chemotherapy. However, although these methods can improve the survival rate of patients to a certain extent, many patients still experience relapse and no response to treatment. On the other hand, the types of drugs currently used in the clinical treatment of leukemia are still limited and it is difficult to meet the growing clinical needs. Therefore, there is an urgent need to develop more anti-leukemia drugs, especially drugs with more efficient targeting and stronger cancer cell membrane penetration ability, to provide more options for medical research and clinical applications. 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, wherein the tetrahydroisoquinoline compound has a structure of formula (I):

[0009]

[0010] in, 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, hydroxyl or amino;

[0012] The tetrahydroisoquinoline compounds of the present invention can, on the one hand, target and inhibit the interaction between nuclear receptor coactivator factor 4 and ferritin heavy chain, inhibit iron autophagy, thereby inhibiting 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 The targeting and ability to penetrate cancer cell membranes of drugs can be improved, thereby increasing the activity of the compound. Since the surface of cancer cells usually carries a weak negative charge, the amide molecules with positive charges are more likely to reach the surface of cancer cells and be absorbed, and have a high anti-cancer effect. Therefore, the tetrahydroisoquinoline compounds of the present invention can provide more options for medical research and clinical applications.

[0013] Furthermore, the is a substituted amide group formed by the carboxylic acid and the amino group on 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, 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 of an alkoxy group;

[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 the amino group on any one of 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 one 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 For the mono-substituted piperazine, 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, and dry N,N-dimethylformamide (9-11 mL) was added, and the mixture was stirred at room temperature for 50-70 min, followed by compound 3 (0.8-1.2 mmol), and the mixture was stirred under magnetic stirring at 60° C. for 11-13 hours, and then extracted with ethyl acetate (3×150 mL, i.e., 150 mL of ethyl acetate was used for extraction three times), the organic layer was dried over anhydrous sodium sulfate, and concentrated under reduced pressure, and the solid obtained by spin drying was separated and purified by column chromatography to obtain a yellow solid, thereby obtaining 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, thereby obtaining 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 spin-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 agent, an injection, an external agent or an inhalation agent.

[0031] Furthermore, the oral agent 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, can enhance the activity of the compounds, and have high anti-cancer effects;

[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 drawings required for use in the embodiments are briefly introduced below. It should be understood that the following drawings only show certain embodiments of the present invention and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other related drawings can be obtained based on these drawings without creative work.

[0037] Figure 1 This is a statistical graph of 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 of the effects of compounds 1a-1j in Examples 1-10 and compound 9 in Comparative Example 1 on the clone 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 The acute toxicity test results of compound 1a in Example 1 are shown. DETAILED DESCRIPTION

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

[0043] Example

[0044] The present invention is further described below in conjunction with specific examples and comparative examples. The following specific examples are all 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 embodiments 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, and the mixture was stirred at room temperature for 1 hour. Subsequently, 6-amino-1,2,3,4-tetrahydroisoquinoline-2-carboxylic acid-2-methylprop-2-yl ester (1.0 mmol) was added, and the mixture was stirred magnetically 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, thereby obtaining 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] 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] - .

[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, finely filtered, and sterilized by filling according to conventional methods 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 in 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 from ATCC.

[0093] Cell culture: cells were cultured in a 37°C, 5% CO2 incubator. After being taken out of the cryopreservation tank, the cells were thawed as quickly as possible in a 37°C water bath and immediately placed in IMDM medium amended with 20% fetal bovine serum and 10 μg / mL DNASE I. The cells were centrifuged at 1500 rpm for 5 minutes and then resuspended in complete medium at a concentration of 2-5×10 6 cells / mL; the thawed cells were then cultured in complete medium, IMDM medium supplemented with 10% fetal bovine serum (FBS) and BIT (bovine serum albumin 4g / L, insulin 5μg / mL, transferrin 60μg / mL, all from Sigma-Aldrich), in order to promote cell growth and maintenance, some specific cytokines and components were added: 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), in addition, the culture medium also contained 50μM β-mercaptoethanol (Sigma-Aldrich).

[0094] Test method: L-CFU method was used to detect clone formation, and cells were counted at 1×10 5 / mL was inoculated in H4230 culture medium, and the culture medium was 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). They were divided into control group and drug treatment group, and the drug treatment concentration was 1μM. On the 7th day, 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 leukemia cell clone formation ( Figure 1 and Figure 2 ), and the compounds of the present invention have stronger anticancer effects than compound 9. This also proves that after the introduction of the amide group, the ability to penetrate the cell membrane is enhanced, which improves the activity of the compound, especially 1a, 1d, 1e and 1f have extremely obvious effects on reducing the formation of leukemia cell clones, among which 1a has the best comprehensive effect.

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

[0098] 1. Experimental methods

[0099] Intracellular Fe 2+ Level determination: HL-60 and Kasumi-1 cells were seeded in 96-well laser confocal culture plates, with 5000 cells in 100 μL culture medium per well. 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 then 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 a 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 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 mouse feed and sterile water. After one week of breeding in the animal room after purchase, no abnormalities were observed. They were randomly divided into 4 groups (control group, 100 mg / kg, 200 mg / kg, 400 mg / kg), 6 mice in each group, half male and half female, and fed in separate cages at the same time.

[0105] The mice were fasted for 12 hours before administration and allowed free access to water. The mice were then given the drug by gavage 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 behaviors and deaths 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 of 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 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 principle of the present invention should be included in the protection scope 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 a structure of formula (I): in, 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; 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 an alkoxy group, a hydroxy group or an amino group.

2. The use according to claim 1, characterized in that: Said is a substituted amide group formed by the carboxylic acid and the amino group on 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, 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; The R 211 is a substituted piperazine, wherein the substituent of the substituted piperazine is C 1～3 alkyl.

3. The use according to claim 2, characterized in that: Said is a substituted amide group formed by the carboxylic acid and the amino group on any one of 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 one of the alkoxy groups.

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

5. The use according to claim 1, characterized in that: The tetrahydroisoquinoline compound can be used in the form of any one of its pharmaceutically acceptable salts, solvates or chiral isomers.

6. The use according to claim 5, characterized in that: The pharmaceutically acceptable salt is obtained by reacting a tetrahydroisoquinoline compound with an inorganic acid or an organic acid.

7. The use 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, 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.

8. The use 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 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.

10. The use according to claim 1, characterized in that: The oral preparation is any one of capsules, tablets, pills and granules; the inhalation agent is a spray.

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