CN105796638B - Application of oridonin and cryptotanshinone in preparation of leukemia treatment drug - Google Patents

Application of oridonin and cryptotanshinone in preparation of leukemia treatment drug Download PDF

Info

Publication number
CN105796638B
CN105796638B CN201610336683.7A CN201610336683A CN105796638B CN 105796638 B CN105796638 B CN 105796638B CN 201610336683 A CN201610336683 A CN 201610336683A CN 105796638 B CN105796638 B CN 105796638B
Authority
CN
China
Prior art keywords
cryptotanshinone
oridonin
leukemia
treating
drug
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Active
Application number
CN201610336683.7A
Other languages
Chinese (zh)
Other versions
CN105796638A (en
Inventor
董博文
周惠
屈良鹄
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Sun Yat Sen University
Original Assignee
Sun Yat Sen University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Sun Yat Sen University filed Critical Sun Yat Sen University
Priority to CN201610336683.7A priority Critical patent/CN105796638B/en
Publication of CN105796638A publication Critical patent/CN105796638A/en
Application granted granted Critical
Publication of CN105796638B publication Critical patent/CN105796638B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Classifications

    • 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
    • A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
    • A61K31/58—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids containing heterocyclic rings, e.g. danazol, stanozolol, pancuronium or digitogenin
    • 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
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/53—Lamiaceae or Labiatae (Mint family), e.g. thyme, rosemary or lavender
    • A61K36/537—Salvia (sage)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs

Landscapes

  • Health & Medical Sciences (AREA)
  • Natural Medicines & Medicinal Plants (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Veterinary Medicine (AREA)
  • Epidemiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Chemical & Material Sciences (AREA)
  • Botany (AREA)
  • Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Alternative & Traditional Medicine (AREA)
  • Medical Informatics (AREA)
  • Microbiology (AREA)
  • Mycology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

The invention discloses an application of oridonin and cryptotanshinone in preparing a medicament for treating leukemia. The research of the invention finds that the combined medication of quantitative compatibility of the effective components of the Chinese herbal medicines with different anticancer mechanisms, namely the oridonin and the cryptotanshinone, can generate obvious synergistic effect. The compatibility mode can kill more leukemia cells by using the drug combination with lower concentration, and can effectively reduce the total concentration of the drugs corresponding to the leukemia cell proliferation inhibition rate of the drugs reaching 50 percent, and the dosage of each drug in the compatibility mode does not belong to the range of causing harm to organisms. Therefore, the combined medication scheme of the oridonin and the cryptotanshinone has important significance and wide application prospect in the aspects of preparing leukemia treatment medicines and preventing and researching leukemia.

Description

Application of oridonin and cryptotanshinone in preparation of leukemia treatment drug
Technical Field
The invention belongs to the technical field of biological medicines. More particularly, relates to the application of oridonin and cryptotanshinone in preparing drugs for treating leukemia.
Background
Leukemia, commonly known as "leukemia", is a type of hematological malignancy derived from hematopoietic stem cells. Currently, treatment of leukemia is mainly focused on the following protocols: chemotherapy, radiation therapy, bone marrow transplantation, immunotherapy, and the like. Among them, chemotherapy is the mainstream of leukemia therapy because of its relatively strong targeting and low treatment cost.
In the common chemotherapy scheme, the idea of drug combination is fully embodied. At present, the common clinical combined administration mode mainly comprises western medicines combined with western medicines and western medicines combined with traditional Chinese medicines, the western medicines are mostly specific or non-specific blocking medicines for cell cycle, and the abnormal proliferation characteristic of leukemia cells is utilized to treat the disease.
However, the above-mentioned therapies also have significant problems in application: first, chemotherapy based on cell cycle blockers prevents abnormal proliferation of leukemic cells, but also prevents normal body cell division; secondly, the metabolism process of western medicines is complex, and the western medicines seriously damage the heart, the liver, the kidney and other important organs of a patient; thirdly, the price of the targeted chemotherapy drug is not so high that the targeted chemotherapy drug is difficult to bear by ordinary families for a long time.
Disclosure of Invention
The invention aims to solve the technical problems of overcoming the defects and shortcomings of the existing drugs for treating leukemia, mainly overcoming the defects of poor targeting property (namely normal cells and cancerated cells are not distinguished, and proliferation of all cells of an organism is forcibly retarded) of a chemotherapy scheme, strong side effect, high cost and the like, providing a combined medication method for matching two effective Chinese herbal medicine components of Oridonin (Oridonin for short) and Cryptotanshinone (CPT for short) with different anticancer mechanisms, achieving the aim of killing leukemia cells with low dose and high efficiency by utilizing the obvious synergistic effect between the two compounds, and greatly reducing the toxic and side effects of the drugs on the organism in the treatment process and remarkably reducing the treatment cost due to low toxicity and rich sources of the Chinese herbal medicines and easily obtained materials.
The invention aims to provide application of a combined medication scheme of oridonin and cryptotanshinone in preparing a medicament for treating leukemia.
The invention also aims to provide a medicament for treating leukemia.
The above purpose of the invention is realized by the following technical scheme:
application of Rabdosia rubescens and Salvia miltiorrhiza in preparation of drugs for preventing, treating, adjunctively treating and/or prognostically preventing recurrence of leukemia.
Application of oridonin and cryptotanshinone in preparing medicine for preventing, treating, adjunctively treating and/or prognosing and preventing recurrence of leukemia is provided.
In particular to the application of a high-efficiency and low-toxicity medicine composition prepared by combining oridonin and cryptotanshinone according to a certain proportion in the preparation of medicines for preventing, treating, assisting in treating and/or prognosing and preventing relapse of leukemia.
The oridonin or cryptotanshinone comprises pharmaceutically acceptable salts and esters thereof, selectively substituted analogues or a combination of one or more of the foregoing compounds; also includes the derivatives of the two, or the pharmaceutically acceptable salts or solvates of the two and the derivatives thereof.
The medicine has the advantages of high efficiency and low toxicity.
Preferably, the leukemia is chronic myelogenous leukemia or acute myelogenous leukemia.
More preferably, the leukemia is chronic myelogenous leukemia cell line K562 cells or acute myelogenous leukemia cell line HL60 cells.
The active ingredients with toxicity reducing and efficacy enhancing effects in the combined compatibility scheme are rubescensin A and cryptotanshinone.
Preferably, in the application scheme, the molar concentration ratio of the oridonin to the cryptotanshinone is 0.1-15: 0.1 to 20.
Preferably, when the medicine for treating chronic myelogenous leukemia is prepared, the molar concentration ratio of the oridonin to the cryptotanshinone is 5-10: 3 to 7.5; when the medicine for treating acute myelogenous leukemia is prepared, the molar concentration ratio of the oridonin to the cryptotanshinone is 1-2.5: 2.5 to 5.
More preferably, when preparing the medicament for chronic myelogenous leukemia, the molar concentration ratio of oridonin to cryptotanshinone is 5: 7.5 or 10: 3; when preparing the medicine for acute myeloid leukemia, the molar concentration ratio of the oridonin to the cryptotanshinone is 1: 5 or 2.5: 2.5.
in addition, preferably, the concentrations of the combination of the oridonin and the cryptotanshinone are respectively as follows: the concentration of the oridonin is less than 15 muM, and the concentration of the cryptotanshinone is less than 20 muM.
More preferably, the concentration of the oridonin is less than 10 muM, and the concentration of the cryptotanshinone is less than 20 muM.
Most preferably: when preparing the medicine for the chronic myelogenous leukemia, the optimal compatibility scheme is that 5 muM oridonin is compatible with 7.5 muM cryptotanshinone, or 10 muM oridonin is compatible with 3 muM cryptotanshinone; when preparing the drug for acute myelogenous leukemia, the optimal compatibility scheme is that 1 muM oridonin is compatible with 5 muM cryptotanshinone, or 2.5 muM oridonin is compatible with 2.5 muM cryptotanshinone.
Research results show that when the oridonin 5 muM is compatible with the cryptotanshinone 7.5 muM or the oridonin 10 muM is compatible with the cryptotanshinone 3 muM, the proliferation inhibition rate of the pharmaceutical composition on chronic myelogenous leukemia cells can reach 50%; when 1 mu M of rubescensin A is compatible with 5 mu M of cryptotanshinone or 2.5 mu M of rubescensin A is compatible with 2.5 mu M of cryptotanshinone, the proliferation inhibition rate of the pharmaceutical composition on acute myelogenous leukemia cells can reach 50%; and the use amount of the two compatibility modes does not belong to the range of harm to organisms.
A pharmaceutical composition for treating leukemia comprises effective amounts of Rabdosia Rubescens (Hemsl.) Hara and Saviae Miltiorrhizae radix, or comprises effective amounts of oridonin and cryptotanshinone. The medicament can also be a composition containing oridonin or cryptotanshinone derivatives, pharmaceutically acceptable salts and esters thereof, selectively substituted analogues or one or more of the compounds.
Furthermore, the pharmaceutical composition can also comprise acceptable pharmaceutical carriers to prepare various pharmaceutically acceptable preparations, including injection or freeze-dried powder injection preparations, pills, decoctions, tablets, granules, hard capsules, soft capsules, controlled release, sustained release preparations, injections or oral preparations.
Such pharmaceutically acceptable carriers include (but are not limited to): saline, buffer solution, glucose, water, glycerol, ethanol, low molecular weight dextran, polyethylene glycol 400, polyethylene glycol 6000, mannitol, lactose, glucose, sucrose, sodium chloride and/or sorbitol.
The preparation method of the medicament consisting of oridonin and cryptotanshinone is not restricted strictly, and can be prepared into injection forms, for example, physiological saline or aqueous solution containing glucose and other auxiliary agents by a conventional method. Pharmaceutical compositions, such as tablets and capsules, can be prepared by conventional methods. Pharmaceutical compositions such as injections, solutions, tablets and capsules are preferably manufactured under sterile conditions.
Furthermore, the pharmaceutical composition may further comprise more than one pharmaceutically acceptable excipient.
In addition, the pharmaceutical composition can also comprise other chemotherapeutic drugs, anti-inflammatory drugs, hormone drugs, small molecule targeted drugs, antibody drugs, Chinese medicinal compound preparations, Chinese medicinal extracts and/or Chinese medicinal monomers and derivatives thereof, so as to prepare drugs with different pertinences.
Particularly preferably, the other chemotherapeutic or anti-cancer agent may be (but is not limited to) one or more of the following: etoposide, 5-fluorouracil, cyclophosphamide, doxorubicin, daunorubicin, epirubicin, cytarabine, isatin, cisplatin, carboplatin, etoposide, topotecan, irinotecan, and the like.
The anti-inflammatory drug or immune stimulant may be (but is not limited to) one or more of the following: aspirin, interferon, c-AMP, globulin, and the like.
The hormonal agent may be (but is not limited to) one or more of the following: glucocorticoids, estrogens, progestins, and the like.
The small molecule targeted drug may be (but is not limited to) one or more of the following: imatinib (Imatinib), Dasatinib (Dasatinib), Nilotinib (Nilotinib), Bosutinib (Bosutinib), Sunitinib (Sutent, Sunitinib), Sorafenib (Nexavar, Sorafenib), Lapatinib (Lapatinib), iressa (Gefitinib), and the like.
The antibody-based drug may be (but is not limited to) one or more of the following: rituxan (Rituximab), Herceptin (Trastuzumab, Herceptin), Erbitux (Cetuximab, Erbitux), Bevacizumab (Avastin ), Panitumumab (Panitumumab), Nimotuzumab (Nimotuzumab), and the like.
The anti-tumor traditional Chinese medicine compound preparation and the traditional Chinese medicine extract can be one or more of (but not limited to): ganoderma extract, Coriolus versicolor extract, Poria extract, Coicis semen extract, radix Dendropanacis mixture, Bufonis venenum, etc.
The traditional Chinese medicine monomer and the derivative thereof can be (but are not limited to) one or more of the following: paclitaxel, camptothecin, homoharringtonine, artemisinin, soybean isoflavone, ginsenoside, vincristine, colchicine, naringenin, emodin and its derivatives, matrine and its derivatives, triptolide and its derivatives, etc.
In addition, the application of the pharmaceutical composition in the preparation of attenuated synergistic anti-leukemia drugs, drugs for preventing leukemia, drugs for improving immunity of the organism, health-care drugs or health-care foods and the drugs prepared from the pharmaceutical composition are also within the protection scope of the invention.
Wherein, preferably, the leukemia is chronic myelogenous leukemia or acute myelogenous leukemia.
More preferably, the leukemia is chronic myelogenous leukemia cell line K562 cells or acute myelogenous leukemia cell line HL60 cells.
The invention provides a combined medication mode for matching the effective components of the Chinese herbal medicines with different anti-cancer mechanisms, namely the oridonin and the cryptotanshinone, and provides a specific matching dose and the inhibition situation of cell proliferation under the corresponding dose, the matching mode can greatly improve the proliferation inhibition effect of the medicine on leukemia cells, and the total concentration of the medicine is obviously reduced when the proliferation inhibition rate of the medicine on the leukemia cells reaches 50 percent.
The invention also determines that the pharmaceutical composition (the oridonin and the cryptotanshinone are compatible) has obvious synergistic effect under the condition of compatibility of certain dosage, so that the pharmaceutical composition can be applied to clinical treatment, auxiliary treatment and prognosis recurrence prevention related treatment of leukemia at relatively low dosage, and the compatibility of the pharmaceutical composition improves the total curative effect of the pharmaceutical composition and reduces the occurrence of side effects. The invention discloses a pharmaceutical composition containing the compound, a relevant proportion thereof and application of the pharmaceutical composition in the aspect of efficiently and low-toxicity prevention and treatment of leukemia.
The invention has the following beneficial effects:
the invention discloses a high-efficiency low-toxicity treatment of leukemia by the compatibility and combination of effective Chinese herbal medicine components with different anticancer mechanisms, which is mainly characterized in that the effective components of rubescensine A and cryptotanshinone in the Chinese herbal medicine rabdosia rubescens and salvia miltiorrhiza are subjected to quantitative compatibility and combination, thereby generating obvious synergistic effect.
In the compatibility modes (oridonin and cryptotanshinone), the combined use of the oridonin and the cryptotanshinone can play a good synergistic effect, the medicine combination can obviously reduce the total concentration of the corresponding medicines when the leukemia cells reach half of the proliferation inhibition rate (namely, the total concentration of the corresponding medicines when the leukemia cell proliferation inhibition rate of the medicines reaches 50 percent can be effectively reduced), and the use amount of the two compatibility modes does not belong to the range of damaging organisms.
Therefore, the combined drug scheme of the oridonin and the cryptotanshinone has important significance and wide application prospect in the aspects of preparing leukemia treatment drugs and preventing and researching leukemia.
Drawings
FIG. 1 shows the proliferation inhibition of cells of a chronic myelogenous leukemia cell strain K562 by combining 5 mu M oridonin and 10 mu M oridonin with drug compatibility modes of different concentrations of cryptotanshinone.
FIG. 2 shows the proliferation inhibition of cells of acute myeloid leukemia cell line HL60 by the drug compatibility of 1 muM and 2.5 muM oridonin in combination with different concentrations of cryptotanshinone.
Detailed Description
The invention is further described with reference to the drawings and the following detailed description, which are not intended to limit the invention in any way. Reagents, methods and apparatus used in the present invention are conventional in the art unless otherwise indicated.
Unless otherwise indicated, reagents and materials used in the following examples are commercially available.
Example 1 study of combined application of oridonin and cryptotanshinone in chronic myelogenous leukemia cell lines
Selecting oridonin and cryptotanshinone with certain concentration gradient to treat chronic myelogenous leukemia K562 cells for 24 hours respectively, fixing the concentration of the oridonin serving as an anticancer drug, treating the leukemia K562 cells for 24 hours by jointly using cryptotanshinone with different gradient concentrations, and calculating the respective proliferation inhibition strength of the cells in three different drug treatment modes by using an MTT colorimetric method.
The specific research method is as follows:
1. experimental cell lines
K562 chronic myelogenous leukemia cells were obtained from the bioengineering research center of zhongshan university (guangdong, guangzhou).
2. Cell culture:
k562 cells were cultured in a conventional cell culture medium (RPMI-1640 medium containing 10% FBS and 1% double antibody) at a constant temperature of 37 deg.C and 5% CO2The conditions of (1) are carried out.
3. Drug mother liquor preparation experiment:
oridonin powder from Henan Jishi pharmaceuticals, Inc. and Cryptotanshinone powder from Selleck Chemicals, Inc. were weighed and dissolved in DMSO solutions to final concentrations of 40mM (ori) and 5mM (CPT), respectively.
4. Cell proliferation inhibition assay:
(1) obtaining cells
Collecting leukemia K562 cells cultured to logarithmic growth phase, centrifuging at 750rpm for 3 min, suspending the cells in RPIM 1640 medium containing 10% fetal calf serum, and adjusting the concentration of the cell suspension to 3 × 105Inoculating the cell suspension into a 96-well plate at a rate of 98 muL/well, and placing at 37 ℃ and 5% CO2Culturing in an incubator for 24 hours;
(2) adding chemicals for treatment
A. Diluting the prepared 40mM oridonin mother liquor to 100 times of working concentration (5 mu M and 10 mu M) by using a DMSO solution, so that the preparation concentration is 0.5mM and 1mM respectively, and uniformly mixing by blowing;
B. diluting the prepared 5mM cryptotanshinone mother liquor with a DMSO solution to 100 times of working concentrations (2.5 muM, 5 muM, 7.5 muM, 10 muM and 20 muM), so that the preparation concentrations are respectively 0.25mM, 0.5mM, 0.75mM, 1mM and 2mM, and uniformly mixing by blowing;
C. adding diluted medicines into cell suspension of each hole in the 96-hole plate in the step (1), wherein the medicines are divided into a single cryptotanshinone medicine group, a 5 mu M combined medicine group of the oridonin and the cryptotanshinone, and a 10 mu M combined medicine group of the oridonin and the cryptotanshinone:
the cryptotanshinone is used independently: adding 1 mu L B diluted cryptotanshinone solution and 1 mu LDMSO solution at each concentration into each well, adding 2 mu L DMSO solution into 0 mu M control well of cryptotanshinone, shaking the culture plate to mix well, placing at 37 ℃, and placing at 5% CO2Culturing in an incubator for 24 hours;
5 mu M combined drug combination of rubescensine A and cryptotanshinone: adding 0.5mM rubescensin A solution diluted in 1 mu L A and cryptotanshinone solutions diluted in 1 mu L B into each well, adding 0.5mM rubescensin A solution diluted in 1 mu L A and 1 mu L DMSO solution into control wells of 5 mu M cryptotanshinone A, shaking the culture plates to mix uniformly, placing at 37 ℃, and placing at 5% CO2Culturing in an incubator for 24 hours;
10 mu M of combined drug combination of rubescensine A and cryptotanshinone: adding 1mM rubescensin A solution diluted in 1 mu L A and cryptotanshinone solutions diluted in 1 mu L B into each well, adding 1mM rubescensin A solution diluted in 1 mu L A and 1 mu L DMSO solution into 10 mu M cryptotanshinone 0 mu M control wells of rubescensin A, shaking the culture plates to mix uniformly, placing at 37 ℃, and placing at 5% CO2Culturing in an incubator for 24 hours;
(3) to each well of cell suspension was added 15. mu.L of MTT solution (MTT kit available from Promega, USA) and the mixture was placed at 37 ℃ with 5% CO2Culturing for 4 hours in an incubator;
(4) adding 100 mu L STOP solution into each hole in the step (3), placing at 37 ℃ and 5% CO2Culturing in an incubator for 24 hours;
(5) taking out the 96-well culture plate, and detecting the absorbance of each well under excitation light with wavelength of 570nm and 630nm, respectively, A570-A630The real light absorption value of each well is obtained, the survival rate of the cells in each well is reflected, and the proliferation inhibition condition of the cells in each well under the action of the medicament can be calculated according to the survival rate.
5. Results of the experiment
The result is shown in figure 1, and the combined use of oridonin and cryptotanshinone can play a significant synergistic effect.
The medicine composition can obviously reduce the total concentration of the corresponding medicines when the leukemia cells reach half of the proliferation inhibition rate: the oridonin 5 muM is compatible with the cryptotanshinone 7.5 muM (shown in figure 1) or the oridonin 10 muM is compatible with the cryptotanshinone 3 muM (shown in figure 1), and the use amount of the two compatibility modes does not belong to the range capable of causing harm to organisms.
Example 2 research of combined application of oridonin and cryptotanshinone in acute myelogenous leukemia cell strain
Selecting oridonin and cryptotanshinone with certain concentration gradient to treat HL60 cell for 24 hr, fixing the concentration of oridonin as anticancer medicine, treating HL60 cell for 24 hr, and calculating the respective proliferation inhibiting strength of the cell in three different medicine treating modes via MTT colorimetry.
The specific research method is as follows:
1. experimental cell lines
HL60 acute myeloid leukemia cells were obtained from the bioengineering research center of the university of zhongshan (guangdong, guangzhou).
2. Cell culture:
culturing HL60 cells with whole culture medium (RPMI-1640 culture medium containing 10% FBS and 1% double antibody) at 37 deg.C under 5% CO2The conditions of (1) are carried out.
3. Drug mother liquor preparation experiment:
oridonin powder from Henan Jishi pharmaceuticals, Inc. and Cryptotanshinone powder from Selleck Chemicals, Inc. were weighed and dissolved in DMSO solutions to final concentrations of 40mM (ori) and 5mM (CPT), respectively.
4. Cell proliferation inhibition assay:
(1) obtaining cells
Collecting leukemia HL60 cells cultured to logarithmic growth phase, centrifuging at 750rpm for 3 min, suspending the cells in RPIM 1640 medium containing 10% fetal bovine serum, and adjusting the concentration of the cell suspension to 3 × 105Inoculating the cell suspension into a 96-well plate at a rate of 98 muL/well, and placing at 37 ℃ and 5% CO2Culturing in an incubator for 24 hours;
(2) adding chemicals for treatment
A. Diluting the prepared 40mM oridonin mother liquor with a DMSO solution to 100 times of working concentration (1 mu M and 2.5 mu M), so that the preparation concentration is 0.1mM and 0.25mM respectively, and uniformly blowing and beating;
B. diluting the prepared 5mM cryptotanshinone mother liquor with a DMSO solution to 100 times of working concentrations (2.5 muM, 5 muM and 7.5 muM), so that the preparation concentrations are 0.25mM, 0.5mM and 0.75mM respectively, and uniformly mixing by blowing;
C. adding diluted medicines into cell suspension of each hole in the 96-hole plate in the step (1), wherein the medicines are divided into a single cryptotanshinone medicine group, a 1 mu M combined medicine group of the oridonin and the cryptotanshinone, and a 2.5 mu M combined medicine group of the oridonin and the cryptotanshinone:
the cryptotanshinone is used independently: adding 1 mu L B diluted cryptotanshinone solution and 1 mu LDMSO solution at each concentration into each well, adding 2 mu L DMSO solution into 0 mu M control well of cryptotanshinone, shaking the culture plate to mix well, placing at 37 ℃, and placing at 5% CO2Culturing in an incubator for 24 hours;
1 mu M combined drug combination of rubescensine A and cryptotanshinone: adding 0.1mM rubescensin A solution diluted in 1 mu L A and cryptotanshinone solutions diluted in 1 mu L B into each well, adding 0.1mM rubescensin A solution diluted in 1 mu L A and 1 mu L DMSO solution into control wells of 1 mu M of rubescensin A, shaking the culture plates to mix uniformly, placing at 37 ℃, and placing at 5% CO2Culturing in an incubator for 24 hours;
2.5 mu M combined drug combination of oridonin and cryptotanshinone: adding 0.25mM rubescensin A solution diluted in 1 mu L A and cryptotanshinone solutions diluted in 1 mu L B into each well, and adding 0 mu M cryptotanshinone solution diluted in 1 mu L A into control wells of 0.25mM rubescensin A diluted in 1 mu L AMixing the solution with 1 μ L DMSO solution, shaking the culture plate to mix well, placing at 37 deg.C and 5% CO2Culturing in an incubator for 24 hours;
(3) to each well of cell suspension was added 15. mu.L of MTT solution (MTT kit available from Promega, USA) and the mixture was placed at 37 ℃ with 5% CO2Culturing for 4 hours in an incubator;
(4) adding 100 mu L STOP solution into each hole in the step (3), placing at 37 ℃ and 5% CO2Culturing in an incubator for 24 hours;
(5) taking out the 96-well culture plate, and detecting the absorbance of each well under excitation light with wavelength of 570nm and 630nm, respectively, A570-A630The real light absorption value of each well is obtained, the survival rate of the cells in each well is reflected, and the proliferation inhibition condition of the cells in each well under the action of the medicament can be calculated according to the survival rate.
5. Results of the experiment
The result is shown in figure 2, and the combined use of oridonin and cryptotanshinone can play a significant synergistic effect.
The medicine composition can obviously reduce the total concentration of the corresponding medicines when the leukemia cells reach half of the proliferation inhibition rate: the dosage of the oridonin 1 muM and the cryptotanshinone 5 muM (shown in figure 2) or the dosage of the oridonin 2.5 muM and the cryptotanshinone 2.5 muM (shown in figure 2) are not within the range of harm to organisms.

Claims (5)

1. The application of the rubescensine A and cryptotanshinone in preparing the drugs for preventing, treating, assisting in treating and/or prognosing and preventing relapse of leukemia is characterized in that the rubescensine A comprises the rubescensine A and pharmaceutically acceptable salts or esters thereof; the cryptotanshinone comprises cryptotanshinone, and its pharmaceutically acceptable salt or ester;
wherein when the medicine for treating chronic myelogenous leukemia is prepared, the molar concentration ratio of the oridonin to the cryptotanshinone is 5-10: 3 to 7.5; when the medicine for treating acute myelogenous leukemia is prepared, the molar concentration ratio of the oridonin to the cryptotanshinone is 1-2.5: 2.5 to 5.
2. A pharmaceutical composition for treating leukemia is characterized by comprising effective amounts of oridonin and cryptotanshinone; the oridonin comprises oridonin and pharmaceutically acceptable salt or ester thereof; the cryptotanshinone comprises cryptotanshinone, and its pharmaceutically acceptable salt or ester;
wherein when the medicine for treating chronic myelogenous leukemia is prepared, the molar concentration ratio of the oridonin to the cryptotanshinone is 5-10: 3 to 7.5; when the medicine for treating acute myelogenous leukemia is prepared, the molar concentration ratio of the oridonin to the cryptotanshinone is 1-2.5: 2.5 to 5.
3. The pharmaceutical composition of claim 2, wherein the pharmaceutical composition can be administered orally or by injection, and further comprises pharmaceutically acceptable carriers for preparing various pharmaceutically acceptable preparations, including injection or lyophilized powder preparation, pill, decoction, tablet, granule, hard capsule, soft capsule, controlled release, sustained release preparation or injection.
4. The pharmaceutical composition of claim 2, further comprising one or more pharmaceutically acceptable excipients.
5. The use of the pharmaceutical composition of claim 2 for the preparation of an attenuated, potentiated anti-leukemia drug, a drug for the prevention of leukemia, a drug for enhancing immunity of the body, or a health care drug.
CN201610336683.7A 2016-05-20 2016-05-20 Application of oridonin and cryptotanshinone in preparation of leukemia treatment drug Active CN105796638B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201610336683.7A CN105796638B (en) 2016-05-20 2016-05-20 Application of oridonin and cryptotanshinone in preparation of leukemia treatment drug

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201610336683.7A CN105796638B (en) 2016-05-20 2016-05-20 Application of oridonin and cryptotanshinone in preparation of leukemia treatment drug

Publications (2)

Publication Number Publication Date
CN105796638A CN105796638A (en) 2016-07-27
CN105796638B true CN105796638B (en) 2020-01-14

Family

ID=56451603

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201610336683.7A Active CN105796638B (en) 2016-05-20 2016-05-20 Application of oridonin and cryptotanshinone in preparation of leukemia treatment drug

Country Status (1)

Country Link
CN (1) CN105796638B (en)

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106635641A (en) * 2016-12-22 2017-05-10 阜阳市殿兴农业科技有限公司 Nutritional health care green tea/strawberry fruit wine
CN109864991B (en) * 2019-03-15 2021-07-09 中山大学 Application of cryptotanshinone in preparation of Ph + acute lymphocytic leukemia chemotherapy sensitization drug
CN111467367B (en) * 2020-05-22 2021-08-17 陈炯 Plant monomer composition for inhibiting tumor cell growth and preparation method and application thereof

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1994293A (en) * 2006-08-18 2007-07-11 上海交通大学医学院附属瑞金医院 Application of oridonin in medicine preparation
CN102247340A (en) * 2010-05-20 2011-11-23 上海亚盛医药科技有限公司 Application of apogossypol ketone in preparation of drugs for treating tumor diseases

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1994293A (en) * 2006-08-18 2007-07-11 上海交通大学医学院附属瑞金医院 Application of oridonin in medicine preparation
CN102247340A (en) * 2010-05-20 2011-11-23 上海亚盛医药科技有限公司 Application of apogossypol ketone in preparation of drugs for treating tumor diseases

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
六种中药提取物对白血病细胞株的体外抑制作用研究;单卿卿等;《四川大学学报（医学版）》;20120630;第43卷(第3期);第362-366页 *

Also Published As

Publication number Publication date
CN105796638A (en) 2016-07-27

Similar Documents

Publication Publication Date Title
US9439874B2 (en) Food composition for nourishing, maintaining and cultivating a variety of stem cells and a method for manufacturing the same
CN105963637B (en) Application of cryptotanshinone and curcumin in preparation of tumor treatment medicine
CN109864991B (en) Application of cryptotanshinone in preparation of Ph + acute lymphocytic leukemia chemotherapy sensitization drug
CN105796638B (en) Application of oridonin and cryptotanshinone in preparation of leukemia treatment drug
JP6389958B2 (en) Medicinal use of anti-tumor for rutile pentacyclic triterpene saponins
CN111704594B (en) Artemisia polyanthrene lactone A-S and Artemisia polyanthrin A-G as well as pharmaceutical composition and application thereof
CN103179967A (en) Anti-tumor pharmaceutical composition
CN105833173B (en) Application of oridonin and curcumin in preparation of medicament for treating leukemia
CN1775267A (en) Kaikoujian extract, Its preparing method and use
WO2024007583A1 (en) Use of ginsenoside in combination with pd-1 blocker in preparing medicament against head and neck squamous cell carcinoma
CN103735653A (en) Traditional Chinese medicine extract with anti-tumor activity as well as preparation method and use thereof
CN101156882B (en) Preparation method of pseudo-ginseng protopanoxadiol saponin
US9943560B2 (en) Medical compositions containing liquorice extracts with synergistic effect
CN103919850B (en) A kind of pharmaceutical composition and its application in antineoplastic is prepared
CN104208073A (en) Application of protopanaxadiol to prepare tumor multidrug resistance reversers
CN102688248A (en) Use of bufadienolide compound in preparing medicines for treating oral mucosal malignant tumors
CN101011543A (en) Antineoplastic medicine composition
KR100485936B1 (en) Anticarcinogenic constituents of ginsenoside Rh2 and Rg3
CN111544479B (en) Application of pharmaceutical composition in preparation of medicines for treating cancers
CN104001117A (en) Cervical-cancer-resistant composition and application thereof
CN116478176B (en) Mongolian arteannuin A-K and pharmaceutical composition thereof, and preparation method and application thereof
CN117209462B (en) Bai Lianhao lactone A-U and pharmaceutical composition thereof, and preparation method and application thereof
CN103655822B (en) Anti-tumor drug as well as preparation method and application thereof
CN111184740B (en) Application of zotara in preparation of medicine for enhancing anticancer activity of paclitaxel
CN102370983A (en) Anti-tumor medicine composition and purpose thereof

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant