CN106309431A - Application of Linderolide-H in preparation of medicaments for treating leukemia - Google Patents

Abstract

The invention relates to application of Linderolide H in the preparation of medicaments for treating leukemia and belongs to the field of medicine. The application of Linderolide H in the preparation of medicaments for treating leukemia, the application of Linderolide H in the preparation of medicaments for proliferation of leukemia cells, and the application of Linderolide H in the preparation of medicaments for inducing leukemia cell apoptosis are included. Linderolide H treats leukemia by inhibiting leukemia cell proliferation and inducing leukemia cell apoptosis. The use of Houttuynoid B according to the present invention in the preparation of medicaments for the treatment of leukemia is first disclosed, and the skeletal type is brand-new and its inhibitory activity against leukemia cells is surprising.

Description

Linderolide H application in preparation treatment leukemia medicament

Technical field

The present invention relates to the new application of compound L inderolide H, particularly relate to Linderolide H in preparation treatment Application in leukemia medicament.

Background technology

Acute leukemia is the Clonal malignant disease that a class hematopoietic stem cell is abnormal.Leukaemia in its clone loses Go to be further differentiated into ripe ability and be stuck in cytocerastic different phase.Leukemia in bone marrow and other hemopoietic tissue Other organs and tissue are gathered and infiltrated to a large amount of hypertrophy of cell, makes normal hematopoiesis suppressed simultaneously, and clinical manifestation is anemia, goes out Blood, infection and each organ infiltration's symptom.According to statistics, leukemia accounts for about the 3% of tumor total incidence, is child and youth In modal a kind of malignant tumor.Leukemic sickness rate is in countries in the world, and Europe and North America sickness rate are the highest, and it is dead Rate is 3.2-7.4/10 ten thousand population.Searching can prevent and treat the medicine of acute leukemia and seem the most urgent.

The compound L inderolide H that the present invention relates to be one within 2014, deliver (Qing Liu, et al., Sesquiterpene lactones from the roots of Lindera Strychnifolia.Phytochemistry, 87 (2013) 112 118.) noval chemical compound, this compound has brand-new bone Frame type, has now been found that it can cause food to damage (Qing Liu, et al., Sesquiterpene lactones from The roots of Lindera strychnifolia.Phytochemistry, 87 (2013) 112 118.), for this The bright Linderolide H related to purposes in preparation treatment leukemia medicament belongs to first public, brand-new owing to belonging to Structure type, and it is unexpectedly strong for treatment leukocythemia liveness, does not exist and is provided any enlightenment by other compounds Possibility, possess prominent substantive distinguishing features, be simultaneously used for leukemic preventing and treating and obviously have significantly progressive.

Summary of the invention

Do not find that it has treatment leukocythemia liveness in it is an object of the invention to study according to existing Linderolide H The present situation of report, it is provided that Linderolide H application in preparation treatment leukemia medicament.

Linderolide H is by suppression leukaemia (HL-60 cell) propagation and inducing leukemia cell (HL-60 Cell) apoptosis, leukemia is treated.

Described compound L inderolide H, structure is as shown in formula I:

The Linderolide H that the present invention relates to purposes in preparation treatment leukemia medicament belongs to first public, and And owing to framework types belongs to brand-new framework types, and its cytotoxicity for leukaemia and the life of apoptosis induction Thing activity is unexpectedly strong, and there is not the possibility being provided any enlightenment by other compounds, possesses prominent substantive distinguishing features, It is simultaneously used for leukemic preventing and treating and obviously has the most progressive.

Detailed description of the invention

The preparation method of compound L inderolide H involved in the present invention see document (Qing Liu, et al., Sesquiterpene lactones from the roots of Lindera strychnifolia.Phytochemistry, 87(2013)112–118.)

The present invention is further detailed explanation by the following examples, but protection scope of the present invention is not by concrete real Execute any restriction of example, but be defined in the claims.

Embodiment 1: the preparation of compound L inderolide H tablet involved in the present invention:

Taking 20 g of compound Linderolide H, the customary adjuvant 180 grams of tablet is prepared in addition, mixing, conventional tablet presses Make 1000.

Embodiment 2: the preparation of compound L inderolide H capsule involved in the present invention:

Taking 20 g of compound Linderolide H, the customary adjuvant such as starch 180 grams of capsule is prepared in addition, mixing, dress Capsule makes 1000.

Its pharmaceutically active is further illustrated below by pharmacodynamic experiment.

Test example: Linderolide H suppression HL-60 cell proliferation and HL-60 cells apoptosis

1 material and method

1.1 material RPMI1640 culture medium are purchased from Gibco company；Tetramethyl azo azoles salt (MTT), Rhodamine 123 are Sigma Products；Caspase-3,9 spectrophotometry test kit is purchased from Nanjing triumphant base biology company limited；Other are changed Learn reagent and be domestic analytical pure.

1.2 cells are cultivated people's promyelocytic leukemia cell line HL-60 and are purchased from Chinese Academy of Sciences's Shanghai cell biological research Institute, is incubated in the RPMI1640 culture fluid containing 10% hyclone, penicillin 100kU/L, streptomycin 100mg/L, 37 DEG C, 5%CO2, cultivates in saturated humidity incubator, and trophophase cell of taking the logarithm is tested.

The separation of 1.3 human peripheral blood single nucleus cells (PBMC)

Aseptic collection health donors venous blood, (the 200U/ml heparin sodium that every milliliter of whole blood adds 0.1ml is molten for anticoagulant heparin Liquid), PBS equimultiple dilute blood, 1:4 adds human lymphocyte separation liquid, and 2000rpm is centrifuged 20min, uses nothing after collecting milky white layer Serum RPMI-1640 culture medium 1500rpm10min centrifuge washing 2 times, it is thus achieved that mononuclearcell, trypan blue exclusion rate is more than 95%, it is resuspended in the RPMI-1640 culture medium of 10%FCS standby.

The impact that HL-60 cell and human PBMC are grown by 1.4 Linderolide H

Inoculate HL-60 cell and human PBMC and 96 orifice plates, 2 × 104, every hole cell (100ul).37 DEG C, 5%CO₂Condition After lower cultivation 24h, experimental group once adds 0.625,1.25,2.5,5.0, the Linderolide H of 10.0ug/mL, continue respectively Cultivate 24,48, after 72h, terminate cultivating, if only adding the matched group of culture fluid, each dosage group sets 4 multiple holes.Before experiment terminates 4h, every hole adds 20ul MTT solution (5g/L), continues to hatch 4h, terminates cultivating, and careful suction abandons supernatant, and every hole adds 150uL DMSO, shakes 10min, selects 570nm wavelength, measures each hole absorbance (A) value, repeat on automatic enzyme connection detector Test 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 Assay apoptosis rates

Collect 0,2.5,5.0,10.0ug/mL Linderolide H process 48h respectively organize cell, PBS washs 2 times, adds Enter 70% ethanol of pre-cooling in-20 DEG C of fixing 24h, after centrifuge washing, add 200uL PI dye liquor, 50ul RNase, 4 DEG C of lucifuges Place 20min, 1500rpm and be centrifuged 5min.Adjusting cell concentration is 1 × 105/ml～1 × 106/ml, flow cytometer Analyzing, excitation source is argon laser, and excitation wavelength is 488nm.

1.6 statistical analysis this experimental result applied statistics are analyzed

Software SPSS13.0 process, all experimental results use x ± s to represent, compare employing one factor analysis of variance between group.

2 results

2.1 Linderolide H affect matched group HL-60 cell active growth to what HL-60 cell and PBMC grew, Through 0.625,1.25,2.5,5.0, the HouttuynoidB of 10.0ug/mL process 24,48, after 72h, HL-60 cell grows the most not Slowing down with degree, and in the time, concentration dependent.Under same concentrations, cultivate 72h Linderolide H several to human PBMC Unrestraint effect, compares with HL-60 cell and has significant difference (P < 0.05) (being shown in Table 1).

The table 1 Linderolide H impact (x ± s, n=4) on HL-60 cell proliferation

* p < 0.05**p < 0.01 is compared 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

After the Linderolide H effect 48h of HL-60 cell and variable concentrations, through flow cytometry analysis, result shows All occur in that before G1 phase peak and represent apoptotic hypodiploid peak, dense along with Linderolide H in the range of finite concentration Degree lift-rising is high, and HL-60 apoptosis rate raises, and wherein 10.0ug/mL Linderolide H effect 48h apoptosis rate reaches (22.16 ± 4.87) % (table 2).

Impact (x ± s, n=3) on HL-60 apoptosis rate after table 2 Linderolide H effect 48h

* p < 0.05, * * p < 0.01 is compared with negative control group

Conclusion: Linderolide H can suppress leukaemia's (HL-60 cell) propagation and inducing leukemia cell (HL-60 cell) apoptosis, therefore, it can for treating leukemia.

Claims (1)

1.Linderolide H application in treatment leukemia medicament, described compound L inderol ide H structure such as formula (I) shown in: