WO2023216281A1 - 3-羟基黄酮在制备治疗mll基因重排急性髓系白血病的药物中的应用 - Google Patents

3-羟基黄酮在制备治疗mll基因重排急性髓系白血病的药物中的应用 Download PDF

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WO2023216281A1
WO2023216281A1 PCT/CN2022/093219 CN2022093219W WO2023216281A1 WO 2023216281 A1 WO2023216281 A1 WO 2023216281A1 CN 2022093219 W CN2022093219 W CN 2022093219W WO 2023216281 A1 WO2023216281 A1 WO 2023216281A1
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hydroxyflavone
myeloid leukemia
acute myeloid
mll gene
cells
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曹楠
林东军
刘静
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中山大学附属第七医院(深圳)
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
    • A61K31/352—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methantheline 
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A61P35/02—Antineoplastic agents specific for leukemia

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  • the present invention relates to the field of biotechnology, and in particular to the use of 3-hydroxyflavone in the preparation of drugs for treating MLL gene rearranged acute myeloid leukemia.
  • AML Acute myeloid leukemia
  • MLL-rearranged MLL rearrangements
  • the object of the present invention is to overcome the shortcomings of the above-mentioned prior art and provide the application of 3-hydroxyflavone in the preparation of drugs for treating MLL gene rearranged acute myeloid leukemia.
  • the present invention provides a new use of 3-hydroxyflavone, and It also provides a new way and option for the treatment of acute myeloid leukemia with MLL gene rearrangement.
  • the present invention provides the use of 3-hydroxyflavone in the preparation of medicines for preventing and/or treating acute myeloid leukemia.
  • the acute myeloid leukemia is MLL gene rearranged acute myeloid leukemia.
  • MLL gene rearranged acute myeloid leukemia is referred to as MLLr AML.
  • 3-Hydroxyflavone also known as flavonol, CAS: NO.577-85-5
  • chemical name is 5,7,4'-trihydroxyflavone, which is the most common type of flavonoids and is widely used. Distributed in fruits and vegetables, it has strong anti-cancer and antioxidant effects.
  • 3-hydroxyflavone can be used to selectively inhibit the proliferation and induce apoptosis of MLL gene-rearranged acute myeloid leukemia cell lines; 3-HF selectively causes G0G1 phase arrest of MLLr AML cells. lag, thereby delaying the progression of MLL gene rearranged acute myeloid leukemia.
  • the 3-hydroxyflavone induces apoptosis of MLL gene-rearranged acute myeloid leukemia cells and inhibits proliferation of MLL gene-rearranged acute myeloid leukemia cells.
  • the concentration of 3-hydroxyflavone is 1-10 ⁇ M.
  • the present invention provides a drug for preventing and/or treating MLL gene rearranged acute myeloid leukemia, the active ingredient of which is 3-hydroxyflavone.
  • the medicine also includes pharmaceutical auxiliary materials.
  • the auxiliary materials are any one or more of excipients, diluents, carriers, flavoring agents, binders and fillers. combination of species.
  • the drug can be introduced into the body alone through injection, spraying, nasal instillation, eye instillation, penetration, absorption, physical or chemical mediated methods, and the body includes but is not limited to muscle, intradermal, subcutaneous, intravenous , mucosal tissue; the drug can also be introduced into the body after being mixed or wrapped with other substances.
  • the present invention provides the use of 3-hydroxyflavone in the preparation of drugs for inhibiting the proliferation of MLL gene-rearranged acute myeloid leukemia cells.
  • the present invention provides the use of 3-hydroxyflavone in the preparation of a drug that induces apoptosis of MLL gene rearranged acute myeloid leukemia cells.
  • the present invention has the following beneficial effects:
  • the present invention has discovered through in vivo and in vitro studies that 3-hydroxyflavone inhibits the growth of MLL gene-rearranged acute myeloid leukemia cells, induces their apoptosis and changes their cell cycle ratio; the present invention has verified through animal levels that 3-hydroxyflavone has killing properties in the body The effect of MLL gene rearrangement on acute myeloid leukemia cells, and no obvious side effects on mice, indicating that the use of 3-hydroxyflavone has smaller side effects, good safety and significant application prospects compared with existing chemotherapy drugs. .
  • Figure 1 is a diagram showing the effect of 3-hydroxyflavone (3-HF) on reducing the viability of MLL gene rearranged acute myeloid leukemia (MLLr AML) cells in Example 1.
  • Figure 2 is a graph showing the results of 3-hydroxyflavone (3-HF) inhibiting the proliferation of MLLr AML in Example 2 (Figure 2-A is a representative graph and statistical graph of MOLM13, Figure 2-B is a representative graph and statistical graph of MV411 , Figure 2-C is the representative picture and statistical chart of U937);
  • Figure 3 is a diagram showing the effect of 3-HF on MLLr AML cell cycle arrest in Example 3 (Figure 3-A is a representative diagram and statistical diagram of MOLM13, Figure 3-B is a representative diagram and statistical diagram of MV411, Figure 3 -C is the representative picture and statistical chart of U937);
  • Figure 4 is a graph showing the results of 3-HF inducing apoptosis in MLLr AML cells in Example 4 (Figure 4-A is a representative graph and statistical graph of MOLM13, Figure 4-B is a representative graph and statistical graph of MV411, Figure Picture 4-C is the representative picture and statistical chart of U937);
  • Figure 5 is a graph showing the effect of 3-HF on the growth of xenograft tumors in MV411 mice in Example 5 (Figure 5-A is a representative graph of tumors; Figure 5-B is a representative graph of tumor size);
  • Figure 6 is a graph showing the effect of 3-HF on the progression of leukemia in MLL-AF9 AML mice in Example 6 ( Figure 6-A is a statistical graph of peripheral blood chimerism rate; Figure 6-B is a statistical graph of survival curve; Figure 6-C is a representative graph and statistical graph of bone marrow chimerism rate; Figure 6-D is a representative graph of peripheral blood leukocytes; Figure 6-E is a photo of the liver and histological sections of liver parenchyma; Figure 6-F is the result of liver weight Figure; Figure 6-G is a picture of the spleen and histological section of the spleen; Figure 6-H is a picture of the spleen weight results).
  • Figure 6-A is a statistical graph of peripheral blood chimerism rate
  • Figure 6-B is a statistical graph of survival curve
  • Figure 6-C is a representative graph and statistical graph of bone marrow chimerism rate
  • Figure 6-D is a representative graph of peripheral blood leukocytes
  • Figure 6-E is a photo
  • Grouping 1DMSO group; 2MLLr AML cell lines MOLM13 and MV411 plus 3-HF group with different concentration gradients (1, 5, 10 ⁇ M); 3non-MLLr AML cell line U937 plus 3-HF with different concentration gradients (1, 5, 10 ⁇ M) HF group.
  • Each group has 5 duplicate holes, which are treated for 12, 24, and 48 hours respectively.
  • Detection indicators Detect cell viability through CCK8, and compare the cell viability of DMSO group and 3-HF group.
  • Different groups of cells were prepared into single cell suspensions in a 96-well plate with a density of 5 ⁇ 10 4 /ml. They were cultured in a 37°C incubator for 12, 24, and 48 hours respectively. 10 ⁇ l of CCK8 solution was added to each well, and the culture plate was placed Incubate in the incubator for 4 hours, and measure the absorbance at 450 nm with a microplate reader.
  • 3-HF can reduce the viability of MLLr AML cell lines MOLM13 and MV411; as the concentration of 3-HF increases, the viability of MLLr AML cells decreases significantly; the effect of 3-HF on the non-MLLr AML cell line U937 Vitality has little impact.
  • Grouping 1DMSO group; 2MLLr AML cell lines MOLM13 and MV411 plus 3-HF group with different concentration gradients (1, 5, 10 ⁇ M); 3non-MLLr AML cell line U937 plus 3-HF with different concentration gradients (1, 5, 10 ⁇ M) HF group. Each group was equipped with 3 duplicate holes, which were treated for 24 and 48 hours respectively.
  • Detection indicators Detect cell proliferation by CFSE, and compare the cell proliferation rates of the DMSO group and the 3-HF group.
  • 3-HF can selectively inhibit the proliferation of MLLr AML cells, and as the concentration of 3-HF increases, it can significantly inhibit the proliferation of MLLr AML cells when incubated for 48 hours; 3-HF It has little effect on the proliferation of non-MLLr AML cell line U937.
  • Example 3 In vitro experiment of 3-HF selectively causing GOG1 phase arrest of MLLr AML cells
  • Detection indicators Detect the cell cycle through PI, and compare the cell cycle progress of the DMSO group and the 3-HF group.
  • the cells were seeded in a 12-well plate at a density of 5x10 5 /ml. After treatment at 37°C, the cells were collected by centrifugation. Add 70% ethanol and place at 4°C overnight to fix the cells. After removing the ethanol, use RNASEA to neutralize the RNA. Cells were stained by PI for 30 min at 4°C before flow cytometric detection.
  • Example 4 In vitro experiment of 3-HF selectively inducing apoptosis of MLLr AML cells
  • Detection indicators Detect cell apoptosis through Annexin V-EV450/7-AAD, and compare the cell apoptosis rates in the DMSO group and the 3-HF group.
  • Different groups of cells were prepared into single cell suspensions in 12-well plates with a density of 5 ⁇ 10 4 /ml, and were cultured in a 37°C incubator for 12, 24, and 48 hours respectively.
  • the treated cells were collected by centrifugation and washed with PBS. , stained with AnnexinV-EV450 and 7-AAD according to supplier specifications, and analyzed using flow cytometry.
  • Grouping 1 Vehicle group: 5% DMSO+55% PEG300+40% NaCl; 2 3-HF 1mg/kg group: 5% 3-HF+55% PEG300+40% NaCl. 4 per group.
  • 5 ⁇ 10 6 MV411 cells were subcutaneously injected into 4- to 6-week-old female BALB/c nude mice. Treatment was initiated when tumor size reached ⁇ 100 mm 3 .
  • the control group 5% DMSO, 55% PEG400, 40% Nacl
  • the treatment group 3-HF 1 mg/kg was intravenously injected once a day. The tumor size was monitored during the treatment, and the mice were sacrificed after 10 days of treatment. Mouse tumors were collected and photographed.
  • mice C57BL/6 mice aged 6 to 8 weeks were irradiated with a sublethal dose (4.5Gy), and 1 ⁇ 10 4 mouse bone marrow cells were injected into the sublethal mice through the tail vein.
  • Mice were randomly divided into two groups 20 days after transplantation and treated with 3-HF or Vehicle for 10 days, with intraperitoneal injection once a day.
  • Flow cytometric analysis of GFP+ cells reflects the level of leukemia cells in the bone marrow.

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Abstract

涉及生物技术领域，具体公开了3-羟基黄酮在制备治疗MLL基因重排急性髓系白血病的药物中的应用。通过体内和体外研究发现，3-羟基黄酮抑制MLL基因重排急性髓系白血病细胞生长，诱导其凋亡并改变其细胞周期比例；通过动物水平验证3-羟基黄酮在体内具有杀伤MLL基因重排急性髓系白血病细胞的作用，而且对小鼠无明显毒副作用，说明采用3-羟基黄酮与现有化疗药物相比具有较小的副作用，安全性好，具有较显著的应用前景。

Description

3-羟基黄酮在制备治疗MLL基因重排急性髓系白血病的药物中的应用 技术领域
本发明涉及生物技术领域，尤其是涉及一种3-羟基黄酮在制备治疗MLL基因重排急性髓系白血病的药物中的应用。
背景技术
急性髓系白血病(Acute Myeloid Leukemia,AML)是成人最常见的白血病类型之一，临床表现主要为大量原始幼稚细胞浸润骨髓导致正常造血功能受抑所致的一系列临床症状，如贫血、出血、感染及组织器官浸润等。经过大量研究者们数十年的研究，目前普遍认为AML是一种起源于造血干祖细(hematopoietic stem/progenitor cells)的恶性克隆增殖性疾病，具有高度的异质性，尤其是伴有混合系白血病(MLL)基因易位的患者多具有化疗不敏感、预后差等特点。临床实践中发现携带MLL重排(MLL-rearranged)患者的预后非常差，治疗反应率约为40％。尽管增加化疗剂量可能降低复发的风险，但也带来长期的不良反应和高的治疗相关死亡率。
因此，迫切需要更有效和毒性更小的药物来治疗MLL基因重排急性髓系白血病。
发明内容
本发明的目的在于克服上述现有技术的不足之处而提供3-羟基黄酮在制备治疗MLL基因重排急性髓系白血病的药物中的应用，本发明提供了3-羟基黄酮的新用途，并且为MLL基因重排急性髓系白血病的治疗也提供了一条新的途径和选择。
为实现上述目的，本发明采取的技术方案为：
第一方面，本发明提供了3-羟基黄酮在制备预防和/或治疗急性髓系白血病的药物中的应用。
作为本发明所述应用的优选实施方式，所述急性髓系白血病为MLL基因重 排急性髓系白血病。为了表达的更加清楚，将MLL基因重排急性髓系白血病简称为MLLr AML。
3-羟基黄酮(3-HF)，又称黄酮醇，CAS：NO.577-85-5，化学名为5,7,4'-三羟基黄酮，是类黄酮中最常见的一类，广泛分布于水果和蔬菜中，具有极强的抗癌和抗氧化作用。
本发明的发明人经过大量研究及试验发现，利用3-羟基黄酮选择性抑制MLL基因重排急性髓系白血病细胞株的增殖及诱导其凋亡；3-HF选择性导致MLLr AML细胞G0G1期阻滞，从而延缓MLL基因重排急性髓系白血病进展。
采用3-羟基黄酮发挥治疗MLL基因重排急性髓系白血病的同时，对小鼠的毒副作用也小，安全性佳。
作为本发明所述应用的优选实施方式，所述3-羟基黄酮诱导MLL基因重排急性髓系白血病细胞凋亡，抑制MLL基因重排急性髓系白血病细胞增殖。
作为本发明所述应用的优选实施方式，所述3-羟基黄酮的浓度为1-10μM。
第二方面，本发明提供了一种预防和/或治疗MLL基因重排急性髓系白血病的药物，所述药物的活性成分为3-羟基黄酮。
作为本发明所述药物的优选实施方式，所述药物还包括药学上的辅料，所述辅料为赋形剂、稀释剂、载体、调味剂、粘合剂和填充剂中的任意一种或多种的组合。
在本发明中，所述药物可单独通过注射、喷射、滴鼻、滴眼、渗透、吸收、物理或化学介导的方法导入机体，所述机体列举但不限于肌肉、皮内、皮下、静脉、粘膜组织；所述药物还能通过被其他物质混合或包裹后导入机体。
第三方面，本发明提供了3-羟基黄酮在制备抑制MLL基因重排急性髓系白血病细胞增殖的药物中的应用。
第四方面，本发明提供了3-羟基黄酮在制备诱导MLL基因重排急性髓系白血病细胞凋亡的药物中的应用。
与现有技术相比，本发明具有如下有益效果：
本发明通过体内和体外研究发现，3-羟基黄酮抑制MLL基因重排急性髓系白血病细胞生长，诱导其凋亡并改变其细胞周期比例；本发明通过动物水平验证3-羟基黄酮在体内具有杀伤MLL基因重排急性髓系白血病细胞的作用，而且对小鼠无明显毒副作用，说明采用3-羟基黄酮与现有化疗药物相比具有较小的 副作用，安全性好，具有较显著的应用前景。
附图说明
图1为实施例1中3-羟基黄酮(3-HF)降低MLL基因重排急性髓系白血病(MLLr AML)细胞活力的影响图。
图2为实施例2中3-羟基黄酮(3-HF)抑制MLLr AML的增殖作用结果图(图2-A为MOLM13的代表图和统计图，图2-B为MV411的代表图和统计图，图2-C为U937的代表图和统计图)；
图3为实施例3中3-HF导致MLLr AML细胞周期阻滞的作用结果图(图3-A为MOLM13的代表图和统计图，图3-B为MV411的代表图和统计图，图3-C为U937的代表图和统计图)；
图4为实施例4中3-HF诱导MLLr AML细胞凋亡的作用结果图(图4-A图为MOLM13的代表图和统计图，图4-B图为MV411的代表图和统计图，图4-C图为U937的代表图和统计图)；
图5为实施例5中3-HF对MV411小鼠异种移植瘤生长的影响结果图(图5-A为肿瘤代表图；图5-B为肿瘤大小的代表图)；
图6为实施例6中3-HF对MLL-AF9 AML小鼠白血病进展的影响结果图(图6-A为外周血嵌合率的统计图；图6-B为生存曲线的统计图；图6-C为骨髓嵌合率的代表图和统计图；图6-D为外周血白细胞的代表图；图6-E为肝脏照片，肝实质的组织学切片；图6-F为肝脏重量结果图；图6-G为脾脏照片，脾脏的组织学切片；图6-H为脾脏重量结果图)。
具体实施方式
为更好的说明本发明的目的、技术方案和优点，下面将结合附图和具体实施例对本发明作进一步说明。
在以下实施例中，所使用的实验方法如无特殊说明，均为常规方法，所用的材料、试剂等，如无特殊说明，均可从商业途径得到。
在以下实施例中，3-羟基黄酮来源于Targetmol。
实施例1、3-羟基黄酮(3-HF)降低MLL基因重排急性髓系白血病(MLLr  AML)细胞活力的作用
(1)实验材料：MLLr AML细胞系MOLM13，MV411，non-MLLr AML细胞系U937。
(2)实验分组情况及培养条件如下：
分组：①DMSO组；②MLLr AML细胞系MOLM13和MV411加不同浓度梯度(1、5、10μM)的3-HF组；③non-MLLr AML细胞系U937加不同浓度梯度(1、5、10μM)的3-HF组。每组设5个复孔，分别处理12，24，48小时。
检测指标：通过CCK8检测细胞活力，比较DMSO组和3-HF组细胞活力。
①比较不同浓度、不同处理时间的3-HF，哪种降低细胞活力的效果最好。
②比较MLLr AML细胞系和non-MLLr AML细胞系，哪种细胞系对3-HF敏感。
(3)实验方法：
不同组细胞在96孔板制成单细胞悬液，密度为5×10 4/ml，放在37℃孵箱分别培养12，24，48小时，每孔加入10μl CCK8溶液，将培养板置于培养箱内孵育4小时，用酶标仪测定在450nm处的吸光度。
(4)结果：
如图1所示，3-HF可以降低MLLr AML细胞系MOLM13，MV411的活力；随着3-HF浓度的增加，MLLr AML细胞的活力明显降低；3-HF对non-MLLr AML细胞系U937的活力影响不大。
实施例2、3-HF选择性抑制MLLr AML细胞增殖的体外实验
(1)实验材料：同实施例1。
(2)实验分组情况及培养条件如下：
分组：①DMSO组；②MLLr AML细胞系MOLM13和MV411加不同浓度梯度(1、5、10μM)的3-HF组；③non-MLLr AML细胞系U937加不同浓度梯度(1、5、10μM)的3-HF组。每组设3个复孔，分别处理24，48小时。
检测指标：通过CFSE检测细胞增殖，比较DMSO组和3-HF组细胞增殖率。
①比较不同浓度、不同处理时间的3-HF对MLLr AML细胞及non-MLLr AML细胞增殖的影响。。
②比较MLLr AML细胞系和non-MLLr AML细胞系，哪种细胞系对3-HF敏 感。
(3)实验方法：
1x10 6/ml细胞悬液在室温下用5μM细胞示踪CFSE染料孵育10分钟。去染料后用PBS洗2次，细胞悬液种在6孔板，并在存在或没有3-HF的情况下孵育24，48h(37℃，5％二氧化碳)。在测量当天，利用流式细胞仪对残余细胞的CFSE荧光进行增殖定量分析。
(4)结果：如图2所示，3-HF可以选择性抑制MLLr AML细胞增殖，并且随着3-HF浓度的增加，当孵育48h时，可以显著抑制MLLr AML细胞的增殖；3-HF对non-MLLr AML细胞系U937的增殖影响不大。
实施例3、3-HF选择性导致MLLr AML细胞G0G1期阻滞的体外实验
(1)实验材料：同实施例1。
(2)实验分组情况及培养条件如下：
分组：同实施例1。每组设3个复孔，分别处理12，24，48小时。
检测指标：通过PI检测细胞周期，比较DMSO组和3-HF组细胞周期进程。
①比较不同浓度、不同处理时间的3-HF对MLLr AML细胞系和non-MLLr AML细胞周期的影响
②比较MLLr AML细胞系和non-MLLr AML细胞系，哪种细胞系对3-HF敏感。
(3)实验方法：
细胞以5x10 5/ml的密度种在12孔板，37℃处理后，离心收集细胞，加70％乙醇置于4℃过夜固定细胞，去乙醇后用RNASEA中和RNA。细胞在4℃下通过PI染色30分钟后进行流式检测。
(4)结果：如图3所述，3-HF选择性导致MLLr AML细胞G0G1期阻滞。
实施例4、3-HF选择性诱导MLLr AML细胞凋亡的体外实验
(1)实验材料：同实施例1。
(2)实验分组情况及培养条件如下：
分组：同实施例1。每组设5个复孔，分别处理12，24，48小时。
检测指标：通过Annexin V-EV450/7-AAD检测细胞凋亡，比较DMSO组和 3-HF组细胞凋亡率。
①比较不同浓度、不同处理时间的3-HF对MLLr AML细胞系和non-MLLr AML细胞凋亡的影响。
②比较MLLr AML细胞系和non-MLLr AML细胞系，哪种细胞系对3-HF敏感。
(3)实验方法：
不同组细胞在12孔板制成单细胞悬液，密度为5×10 4/ml，放在37℃孵箱分别培养12，24，48小时，经过处理的细胞通过离心方式采集，用PBS清洗，根据供应商规格用AnnexinV-EV450和7-AAD进行染色，并使用流细胞学进行分析。
(4)结果：如图4所示，3-HF可以选择性诱导MLLr AML细胞凋亡。
实施例5、3-HF对MV411小鼠异种移植瘤生长的影响
(1)实验材料：MLLr AML细胞系MV411，4-6周龄BALB/c雌性小鼠。
(2)实验分组和处理：
分组：①Vehicle组：5％DMSO+55％PEG300+40％NaCl；②3-HF 1mg/kg组：5％3-HF+55％PEG300+40％NaCl。每组4只。
(3)实验方法：
向4～6周雌性BALB/c裸鼠皮下注射5×10 6个MV411细胞。当肿瘤大小达到～100mm 3开始治疗。对照组(5％DMSO、55％PEG400、40％Nacl)或治疗组(3-HF 1mg/kg)每天腔注射一次，治疗过程中监测肿瘤大小，治疗10天后处死小鼠。收集小鼠肿瘤，拍照。
(4)结果：如图5所示，3-HF在体内实验中对小鼠无明显的毒副作用，并不显著改变小鼠的体重变化，从图5-A可以看出小鼠皮下瘤大小和体积明显小于Vehicle组，具有统计学意义。
实施例6、3-HF在体内阻断MLL白血病的进展
(1)实验材料：MLL-AF9 AML小鼠骨髓细胞，6-8周C57BL/6雌性小鼠。
(2)实验分组和处理：
分组：①Vehicle组：5％DMSO+55％PEG300+40％Nacl；②3-HF 1mg/kg 组：5％3-HF+55％PEG300+40％Nacl。每组5只。
(3)实验方法：
亚致死剂量(4.5Gy)辐照6～8周龄C57BL/6小鼠，用1×10 4个的小鼠骨髓细胞经尾静脉注射至亚致死小鼠。移植后20天将小鼠随机分为两组，3-HF或Vehicle治疗10天，每天腹腔注射1次。通过流式分析GFP+细胞反映了骨髓中白血病细胞的水平。
(4)结果：如图6所示，3-HF可以抑制MLL基因重排急性髓系白血病细胞生长；从图6E-6H可知，3-HF处理组小鼠的肝脾重量及浸润小于Vehicle组。
综上所述，通过体内和体外研究发现，3-HF抑制MLL基因重排急性髓系白血病细胞生长，诱导其凋亡并改变其细胞周期比例；本发明通过动物水平验证3-羟基黄酮在体内具有杀伤MLL基因重排急性髓系白血病细胞的作用，而且对小鼠无明显毒副作用，说明采用3-羟基黄酮与现有化疗药物相比具有较小的副作用，安全性好，具有较显著的应用前景。
最后所应当说明的是，以上实施例仅用以说明本发明的技术方案而非对本发明保护范围的限制，尽管参照较佳实施例对本发明作了详细说明，本领域的普通技术人员应当理解，可以对本发明的技术方案进行修改或者等同替换，而不脱离本发明技术方案的实质和范围。

Claims (8)

  1. 3-羟基黄酮在制备预防和/或治疗急性髓系白血病的药物中的应用。
  2. 如权利要求1所述的应用，其特征在于，所述急性髓系白血病为MLL基因重排急性髓系白血病。
  3. 如权利要求2所述的应用，其特征在于，所述3-羟基黄酮诱导MLL基因重排急性髓系白血病细胞凋亡，抑制MLL基因重排急性髓系白血病细胞增殖。
  4. 如权利要求2所述的应用，其特征在于，所述3-羟基黄酮的浓度为1-10μM。
  5. 一种预防和/或治疗MLL基因重排急性髓系白血病的药物，其特征在于，所述药物的活性成分为3-羟基黄酮。
  6. 如权利要求5所述的药物，其特征在于，所述药物还包括药学上的辅料，所述辅料为赋形剂、稀释剂、载体、调味剂、粘合剂和填充剂中的任意一种或多种的组合。
  7. 3-羟基黄酮在制备抑制MLL基因重排急性髓系白血病细胞增殖的药物中的应用。
  8. 3-羟基黄酮在制备诱导MLL基因重排急性髓系白血病细胞凋亡的药物中的应用。
PCT/CN2022/093219 2022-05-09 2022-05-17 3-羟基黄酮在制备治疗mll基因重排急性髓系白血病的药物中的应用 WO2023216281A1 (zh)

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