CN102872078B - Application of Houttuynoid C in drugs for treating leukemia - Google Patents

Application of Houttuynoid C in drugs for treating leukemia Download PDF

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CN102872078B
CN102872078B CN 201210417571 CN201210417571A CN102872078B CN 102872078 B CN102872078 B CN 102872078B CN 201210417571 CN201210417571 CN 201210417571 CN 201210417571 A CN201210417571 A CN 201210417571A CN 102872078 B CN102872078 B CN 102872078B
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houttuynoid
leukemia
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drugs
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CN102872078A (en
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王慧
张广
吴俊华
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Nantong San Intellectual Property Service Co ltd
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Abstract

The invention relates to an application of Houttuynoid C in drugs for treating leukemia, belonging to the field of medicines. The application of Houttuynoid C in drugs for treating leukemia comprises an application of Houttuynoid C in preparing drugs for restraining multiplication of leukemia cells and application of Houttuynoid C in preparing drugs for inducing apoptosis of leukemia cells. Houttuynoid C treats leukemia by restraining the multiplication of leukemia cells and inducing the apoptosis of leukemia cells. The application of Houttuynoid C in drugs for treating leukemia disclosed by the invention is publicized for the first time; and as the skeleton type is brand new, Houttuynoid C has an unexpectedly strong inhibitory activity to leukemia cells.

Description

The application of Houttuynoid C in the treatment leukemia medicament
Technical field
The present invention relates to the purposes of Houttuynoid C, relate in particular to the application of Houttuynoid C in preparation treatment leukemia medicament.
Background technology
Acute leukemia is the unusual clone's property malignant disease of a class hematopoietic stem cell.Leukaemia among its clone loses the ability of further differentiation and maturation and is stuck in cytocerastic different phase.The a large amount of hypertrophy of leukaemia are gathered and are soaked into other organs and tissue in bone marrow and other hemopoietic tissue, and normal hematopoiesis is suppressed, clinical manifestation be anemia, hemorrhage, infect and each organ soaks into symptom.According to statistics, leukemia accounts for about 3% of tumor total incidence, is modal a kind of malignant tumor among child and the youth.Leukemic sickness rate is in countries in the world, and Europe and North America sickness rate are the highest, and its mortality rate is 3.2-7.4/10 ten thousand populations.The medicine that searching can prevent and treat acute leukemia seems very urgent.
The compound H outtuynoid C that the present invention relates to is one and delivered (Chen in 2012, S. D. et al., 2012. Houttuynoid C_E, Anti-Herpes Simplex Virus Active Flavonoids with Novel Skeletons from Houttuynia cordata. Organic Letters 14 (7), 1772 – 1775.) new framework compound, this chemical compound has brand-new framework types, present purposes only relates to anti-herpes simplex virus activity (Chen, S. D. et al., 2012. Houttuynoid C_E, Anti-Herpes Simplex Virus Active Flavonoids with Novel Skeletons from Houttuynia cordata. Organic Letters 14 (7), 1772 – 1775.), belong to open first for the purposes in preparation treatment leukemia medicament that the present invention relates to, because framework types belongs to brand-new framework types, and its biological activity for leukaemia's cytotoxicity and apoptosis induction is unexpectedly strong, there is not the possibility that is provided any enlightenment by other chemical compounds, possess outstanding substantive distinguishing features, be used for leukemic control simultaneously and obviously have obvious improvement.
Summary of the invention
The invention provides the application of a kind of Houttuynoid C in the leukemic medicine of preparation treatment, to increase the application of Houttuynoid C in medicine.
The application of Houttuynoid C in the leukemic medicine of preparation treatment.
Houttuynoid C comes leukemia is treated by suppressing leukaemia's (HL-60 cell) propagation and inducing leukemia cell (HL-60 cell) apoptosis.
Described compound H outtuynoid C-structure is shown in formula I:
Figure BDA0000231842651
Formula I
The purposes of the Houttuynoid C that the present invention relates in preparation treatment leukemia medicament belongs to open first, and because framework types belongs to brand-new framework types, and its biological activity for leukaemia's cytotoxicity and apoptosis induction is unexpectedly strong, there is not the possibility that is provided any enlightenment by other chemical compounds, possess outstanding substantive distinguishing features, be used for leukemic control simultaneously and obviously have obvious improvement.
The specific embodiment
The preparation method of compound H outtuynoid C involved in the present invention is referring to document (Chen, S. D. et al., 2012. Houttuynoid C_E, Anti-Herpes Simplex Virus Active Flavonoids with Novel Skeletons from Houttuynia cordata. Organic Letters 14 (7), 1772 – 1775.).
The present invention is further detailed explanation by the following examples, but protection scope of the present invention is not subjected to any restriction of specific embodiment, but limited by claim.
Embodiment 1: the preparation of compound H outtuynoid C tablet involved in the present invention:
Get 20 and digest compound Houttuynoid C, add conventional adjuvant 180 grams of preparation tablet, mixing, conventional tablet machine are made 1000.
Embodiment 2: the preparation of compound H outtuynoid C capsule involved in the present invention:
Get 20 and digest compound Houttuynoid C, add conventional adjuvant such as starch 180 grams of preparation capsule, mixing is encapsulatedly made 1000.
Further specify its pharmaceutically active below by pharmacodynamic experiment.
The test example: Houttuynoid C suppresses the HL-60 cell proliferation and induces the HL-60 apoptosis
1 material and method
1.1 material RPMI1640 culture medium is purchased the company in Gibco; Tetramethyl azo azoles salt (MTT), rhodamine 123 are Sigma company product; Caspase-3,9 spectrophotography detection kit are available from the biological company limited of the triumphant base in Nanjing; Other chemical reagent are homemade analytical pure.
1.2 cell culture people promyelocytic leukemia cell strain HL-60 purchases the Shanghai cell biological institute in the Chinese Academy of Sciences, is incubated in the RPMI1640 culture fluid that contains 10% hyclone penicillin 100 kU/L, streptomycin 100 mg/L, 37 ℃, 5%CO 2, to cultivate in the saturated humidity incubator, the trophophase cell of taking the logarithm experimentizes.
1.3 the separation of human peripheral blood single nucleus cell (PBMC)
The healthy donor venous blood of aseptic collection, anticoagulant heparin (every milliliter of whole blood adds the 200U/ml heparin sodium aqua of 0.1ml), PBS etc. times of dilute bloods, 1:4 adds the human lymphocyte separating medium, the centrifugal 20min of 2000 rpm collects serum-free RPMI-1640 culture medium 1500 rpm10min centrifuge washings 2 times of milky white layer back, obtains mononuclearcell, the trypan blue exclusion rate is greater than 95%, is resuspended in the RPMI-1640 culture medium of 10% FCS standby.
1.4 the influence of the HL-60 cell of Houttuynoid C and human PBMC's growth
Inoculation HL-60 cell and human PBMC and 96 orifice plates, every hole 2 * 10 4Individual cell (100ul).37 ℃, 5% CO 2After cultivating 24h under the condition, experimental group once adds the Houttuynoid C of 0.625,1.25,2.5,5.0,10.0 ug/mL, continues to cultivate respectively 24,48, behind the 72h, stops cultivating, and establishes the matched group that only adds culture fluid, and each dosage group is established 4 multiple holes.4h before experiment finishes, every hole adds 20ul MTT solution (5g/L), continue to hatch 4h, stop cultivating, the careful suction abandoned supernatant, every hole adds 150 uL DMSO, concussion 10min selects 570 nm wavelength, measures each hole absorbance (A) value at automatic enzyme connection detector, repeated experiments 3 times, and calculate its growth inhibition ratio (CI).CI=(negative control group A value-experimental group A value)/negative control group A value * 100%.
1.5 flow cytometry is measured apoptosis rate
That collects that 0,2.5,5.0,10.0 ug/mL Houttuynoid C handle 48h respectively organizes cell, PBS washing 2 times, and 70% ethanol that adds pre-cooling is in-20 ℃ of fixing 24h, add 200 uL PI dye liquors behind the centrifuge washing, 50 ul RNA enzymes, 4 ℃ of lucifuges are placed 20 min, the centrifugal 5min of 1500 rpm.Adjusting cell concentration is 1 * 10 5Individual/ml ~ 1 * 10 6Individual/ml, the up flow type cell instrument is analyzed, and excitation source is argon laser, and excitation wavelength is 488 nm.
1.6 this experimental result of statistical analysis applied statistics is analyzed
Software SPSS13.0 handles, and all experimental results adopt x ± s to represent, relatively adopt one factor analysis of variance between group.
2 results
2.1 the influence of the HL-60 cell of Houttuynoid C and PBMC growth
Matched group HL-60 cell active growth, through the Houttuynoid C of 0.625,1.25,2.5,5.0,10.0 ug/mL handle 24,48, behind the 72h, the growth of HL-60 cell is slowing down in various degree all, and be time, concentration dependent.Under same concentrations, cultivate almost unrestraint effect of the 72h human PBMC of Houttuynoid C, with the HL-60 cell significant difference (P<0.05) (seeing Table 1) is arranged relatively.
The influence of the HL-60 cell proliferation of table 1 Houttuynoid C (x ± s, n=4)
Figure BDA0000231842652
Compare * p<0.05 * * p<0.01 with 24h; Compare with 48h P<0.05 △ △P<0.01; Compare with 72h P<0.05 ▲ ▲P<0.01
2.2 Flow cytometry HL-60 apoptosis
Behind the Houttuynoid C effect 48h of HL-60 cell and variable concentrations, through flow cytometry analysis, the result shows G 1All occurred the apoptotic hypodiploid of representative peak before the phase peak, along with Houttuynoid C concentration lift-rising height, the HL-60 apoptosis rate raises, wherein 10.0 ug/mL in the finite concentration scope Houttuynoid C effect 48h apoptosis rate reaches (27.56 ± 4.87) %(table 2).
Behind the table 2 Houttuynoid C effect 48h to the influence of HL-60 apoptosis rate (x ± s, n=3)
Figure BDA0000231842653
Compare * p<0.05, * * p<0.01 with negative control group
Conclusion: Houttuynoid C can suppress leukaemia's (HL-60 cell) propagation and inducing leukemia cell (HL-60 cell) apoptosis, therefore, can be used for leukemia is treated.

Claims (1)

1.Houttuynoid the application of C in preparation treatment leukemia medicament, described compound H outtuynoid C-structure as Formula IShown in:
Figure 41465DEST_PATH_IMAGE001
Formula I.
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Record date: 20231129

Application publication date: 20130116

Assignee: Nantong Yaocheng mechanical equipment manufacturing Co.,Ltd.

Assignor: Nantong San Intellectual Property Service Co.,Ltd.

Contract record no.: X2023980048440

Denomination of invention: The application of Houttuynoid C in the treatment of leukemia drugs

Granted publication date: 20130717

License type: Common License

Record date: 20231129

Application publication date: 20130116

Assignee: Nantong Xinxin Pharmaceutical Co.,Ltd.

Assignor: Nantong San Intellectual Property Service Co.,Ltd.

Contract record no.: X2023980048425

Denomination of invention: The application of Houttuynoid C in the treatment of leukemia drugs

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Application publication date: 20130116

Assignee: Nantong Yijialai Woolen Knitting Co.,Ltd.

Assignor: Nantong San Intellectual Property Service Co.,Ltd.

Contract record no.: X2023980049377

Denomination of invention: The application of Houttuynoid C in the treatment of leukemia drugs

Granted publication date: 20130717

License type: Common License

Record date: 20231203

Application publication date: 20130116

Assignee: NANTONG BEITE PHARMACEUTICAL MACHINERY Co.,Ltd.

Assignor: Nantong San Intellectual Property Service Co.,Ltd.

Contract record no.: X2023980049199

Denomination of invention: The application of Houttuynoid C in the treatment of leukemia drugs

Granted publication date: 20130717

License type: Common License

Record date: 20231203

Application publication date: 20130116

Assignee: NANTONG HAIGUO MACHINERY Co.,Ltd.

Assignor: Nantong San Intellectual Property Service Co.,Ltd.

Contract record no.: X2023980049196

Denomination of invention: The application of Houttuynoid C in the treatment of leukemia drugs

Granted publication date: 20130717

License type: Common License

Record date: 20231203

Application publication date: 20130116

Assignee: Nantong Qiaojiang packaging materials Co.,Ltd.

Assignor: Nantong San Intellectual Property Service Co.,Ltd.

Contract record no.: X2023980049191

Denomination of invention: The application of Houttuynoid C in the treatment of leukemia drugs

Granted publication date: 20130717

License type: Common License

Record date: 20231203

Application publication date: 20130116

Assignee: Nantong Bairun Catering Management Co.,Ltd.

Assignor: Nantong San Intellectual Property Service Co.,Ltd.

Contract record no.: X2023980049185

Denomination of invention: The application of Houttuynoid C in the treatment of leukemia drugs

Granted publication date: 20130717

License type: Common License

Record date: 20231203

Application publication date: 20130116

Assignee: Nantong Nongchaoer Food Co.,Ltd.

Assignor: Nantong San Intellectual Property Service Co.,Ltd.

Contract record no.: X2023980048976

Denomination of invention: The application of Houttuynoid C in the treatment of leukemia drugs

Granted publication date: 20130717

License type: Common License

Record date: 20231203

Application publication date: 20130116

Assignee: Xingren Tank Farm of Nantong Xinhai Liquefied Gas Service Co.,Ltd.

Assignor: Nantong San Intellectual Property Service Co.,Ltd.

Contract record no.: X2023980048969

Denomination of invention: The application of Houttuynoid C in the treatment of leukemia drugs

Granted publication date: 20130717

License type: Common License

Record date: 20231203

Application publication date: 20130116

Assignee: NANTONG SHUANGHE FOOD Co.,Ltd.

Assignor: Nantong San Intellectual Property Service Co.,Ltd.

Contract record no.: X2023980048967

Denomination of invention: The application of Houttuynoid C in the treatment of leukemia drugs

Granted publication date: 20130717

License type: Common License

Record date: 20231203

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Assignee: Jiangsu guanan electromechanical Co.,Ltd.

Assignor: Nantong San Intellectual Property Service Co.,Ltd.

Contract record no.: X2023980048961

Denomination of invention: The application of Houttuynoid C in the treatment of leukemia drugs

Granted publication date: 20130717

License type: Common License

Record date: 20231203

Application publication date: 20130116

Assignee: Nantong Runyuan structural parts Co.,Ltd.

Assignor: Nantong San Intellectual Property Service Co.,Ltd.

Contract record no.: X2023980048952

Denomination of invention: The application of Houttuynoid C in the treatment of leukemia drugs

Granted publication date: 20130717

License type: Common License

Record date: 20231203

Application publication date: 20130116

Assignee: Nantong Jinbao Machinery Manufacturing Co.,Ltd.

Assignor: Nantong San Intellectual Property Service Co.,Ltd.

Contract record no.: X2023980048947

Denomination of invention: The application of Houttuynoid C in the treatment of leukemia drugs

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License type: Common License

Record date: 20231202

Application publication date: 20130116

Assignee: Nantong Juntai motor vehicle testing Co.,Ltd.

Assignor: Nantong San Intellectual Property Service Co.,Ltd.

Contract record no.: X2023980048942

Denomination of invention: The application of Houttuynoid C in the treatment of leukemia drugs

Granted publication date: 20130717

License type: Common License

Record date: 20231202

Application publication date: 20130116

Assignee: Honglei mechanical equipment (Nantong) Co.,Ltd.

Assignor: Nantong San Intellectual Property Service Co.,Ltd.

Contract record no.: X2023980048782

Denomination of invention: The application of Houttuynoid C in the treatment of leukemia drugs

Granted publication date: 20130717

License type: Common License

Record date: 20231202

Application publication date: 20130116

Assignee: American Antelope Home Textile (Nantong) Co.,Ltd.

Assignor: Nantong San Intellectual Property Service Co.,Ltd.

Contract record no.: X2023980048774

Denomination of invention: The application of Houttuynoid C in the treatment of leukemia drugs

Granted publication date: 20130717

License type: Common License

Record date: 20231202

Application publication date: 20130116

Assignee: Nantong Ansheng Textile Co.,Ltd.

Assignor: Nantong San Intellectual Property Service Co.,Ltd.

Contract record no.: X2023980048765

Denomination of invention: The application of Houttuynoid C in the treatment of leukemia drugs

Granted publication date: 20130717

License type: Common License

Record date: 20231130

Application publication date: 20130116

Assignee: NANTONG YA TAI CANDLE ARTS & CRAFTS Co.,Ltd.

Assignor: Nantong San Intellectual Property Service Co.,Ltd.

Contract record no.: X2023980048764

Denomination of invention: The application of Houttuynoid C in the treatment of leukemia drugs

Granted publication date: 20130717

License type: Common License

Record date: 20231130

Application publication date: 20130116

Assignee: Nantong Jianwei Metal Technology Co.,Ltd.

Assignor: Nantong San Intellectual Property Service Co.,Ltd.

Contract record no.: X2023980048759

Denomination of invention: The application of Houttuynoid C in the treatment of leukemia drugs

Granted publication date: 20130717

License type: Common License

Record date: 20231202

Application publication date: 20130116

Assignee: JIANGSU YATAI FITNESS CO.,LTD.

Assignor: Nantong San Intellectual Property Service Co.,Ltd.

Contract record no.: X2023980048748

Denomination of invention: The application of Houttuynoid C in the treatment of leukemia drugs

Granted publication date: 20130717

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Application publication date: 20130116

Assignee: Nantong Zhili Machinery Technology Co.,Ltd.

Assignor: Nantong San Intellectual Property Service Co.,Ltd.

Contract record no.: X2023980049698

Denomination of invention: The application of Houttuynoid C in the treatment of leukemia drugs

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Record date: 20231205

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Application publication date: 20130116

Assignee: JIANGSU HAOHAN INFORMATION TECHNOLOGY Co.,Ltd.

Assignor: Nantong San Intellectual Property Service Co.,Ltd.

Contract record no.: X2024980003183

Denomination of invention: The application of Houttuynoid C in the treatment of leukemia drugs

Granted publication date: 20130717

License type: Common License

Record date: 20240322

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