CN104473946A - Pharmaceutical composition used for treating leukemia - Google Patents

Pharmaceutical composition used for treating leukemia Download PDF

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CN104473946A
CN104473946A CN201410628570.5A CN201410628570A CN104473946A CN 104473946 A CN104473946 A CN 104473946A CN 201410628570 A CN201410628570 A CN 201410628570A CN 104473946 A CN104473946 A CN 104473946A
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pharmaceutical composition
weight portion
cell
leukemia
group
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孔倩倩
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Jinan Xingyi Medical Technology Co Ltd
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Jinan Xingyi Medical Technology Co Ltd
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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
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7028—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
    • A61K31/7032—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a polyol, i.e. compounds having two or more free or esterified hydroxy groups, including the hydroxy group involved in the glycosidic linkage, e.g. monoglucosyldiacylglycerides, lactobionic acid, gangliosides
    • 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
    • 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
    • A61K31/365—Lactones
    • 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

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Epidemiology (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Molecular Biology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Medicines Containing Plant Substances (AREA)

Abstract

The invention relates to a pharmaceutical composition used for treating leukemia. The pharmaceutical composition is prepared by taking morin, podophyllotoxin, sarcostemma acidum aglycone, and soladulcidine as drug substances via mixing. Different dosage forms can be obtained via conventional pharmaceutical technology. The pharmaceutical composition is capable of improving activity of NCK, IL-2, IL-6, and IFN-gamma, and possesses high application value in treatment of leukemia.

Description

One treats leukemic pharmaceutical composition
Technical field
The invention belongs to field of medicaments, relate to one and treat leukemic pharmaceutical composition.
Background technology
Leukemia (Leukemia) is a kind of malignant tumor of hemopoietic system, it is characterized by leukocyte and juvenile cell also has erythroneocytosis or megalokaryocyte (i.e. leukaemia) Progressive symmetric erythrokeratodermia, runaway paraplasm in bone marrow or other hemopoietic tissue, infiltrate the various tissue of whole body and internal organs, and enter in peripheral blood.According to degree and the leukemic natural process of leukaemia's maturation, leukemia mainly can be divided into acute leukemia and the large class of chronic leukemia two.Acute leukemia onset is hurried, causes anemia, hemorrhage, the symptom such as secondary infection and heating because of normocytic minimizing.In addition, respectively organize internal organs because leukaemia extensively infiltrates, also cause liver, spleen, lymphadenectasis and other organ dysfunction.Chronic leukemia onset is delayed, and the course of disease is longer, and early stage patient can without clinical symptoms, the subjective symptoms occurred the earliest often weak, low grade fever, hyperhidrosis, night sweat, the hypermetabolic performance such as lose weight, be found when few patients checks more than health check-up or other.
Leukemia is comparatively common, accounts for about 5% of cancer total incidence, especially with child and youth in the majority.In the mortality rate of each age group malignant tumor of China, leukemia accounts for the 6th (male) and the 8th (women), in the crowd of child and less than 35 years old, then account for the 1st.Leukemic distribution is global
The traditional Chinese medical science is without this noun of leukemia, but it is early on the books in symptoms such as the heating of clinical appearance, anemia, hemorrhage, liver spleen and lymphadenectasis to leukemia, belong to the category such as " empty old ", " anxious old ", " heat is old ", owing to there being hemorrhage also belonging to " blood disorder ", and heating is similar to " epidemic febrile disease ".The obvious person of hepatosplenomegaly, can be described as “ mass in the abdomen " (disorder of YIN and YANG, internal organs are weak, and ailment said due to cold or exposure fights it, be think long-pending), lymphadenectasis obvious person can be described as: " sucutaneous nodule ".Illustrate that motherland's medical science be the dynamic disease of deficiency of YIN fire to leukemic understanding is originally inherent deficient, disorder of YIN and YANG, on the basis of internal organs weakness, caused by ailment said due to cold or exposure pyretic toxicity is taken advantage of a weak point, and clinical manifestation to be deficiency and excess hold assorted general performance under the arm.
Modern leukemic treatment means mainly chemotherapy and bone marrow transplantation therapy.In recent years, along with combined chemotherapy is used for tactful perfect gradually of maintenances, after treatment etc. after clinical and complete incidence graph, and new anti-leukemia medicine is as idarubicin (IDA), mitoxantrone (MIT), the appearance of etoposide (VP-16) and cytosine arabinoside (Ara-C), leukemic treatment level has had breakthrough.But because many unsolved problems such as the toxic and side effects of chemotherapeutics and derived from bone marrow difficulty, force people finding new therapeutic strategy always.Chinese medicine leukemia cause for a long time, is basic rule for the treatment of with heat-clearing and toxic substances removing, supplementing QI and nourishing YIN.Recently increasing research shows, Chinese medicine has obvious advantage and characteristic, for leukemic radical cure provides new Research Thinking and clue in cell death inducing, cell differentiation, reverse leukaemia multidrug resistance and immunomodulating etc.Extract from Chinese medicine, have developed more anti-leukemia medicine and preparation, experiment confirms that these Chinese medicines have antileukemie effect really, and its mechanism of action comprises and suppresses leukaemia DNA, RNA synthesis, promotes apoptosis or differentiation-inducing etc.
Folium et Ramulus Cephalotaxi ester: the harringtonine extracted from Folium et Ramulus Cephalotaxi plant and homoharringtonine, can press down
DNA synthesis processed is a kind of cell cycle nonspecific agent (CCNSA).The HA scheme be made up of harringtonine or homoharringtonine and cytosine arabinoside is widely used in the treatment of acute nonlymphocytic leukemia, and curative effect is better.
Hydroxy camptothecin, hydroxy camptothecin is a kind of Alkaloid of separation and Extraction from China's distinctive plant camptotheca acuminata seed.Mainly suppress nucleic acid especially DNA synthesis in vitro, be S phase specific drug, topoisomerase I is its main site of action.Certain curative effect is had to treatment CML.
Paclitaxel, paclitaxel can be combined with microtubule and microtubule stability is strengthened, and suppresses microtubule depolymerization, makes cell cycle arrest in division stage.Have obvious inhibitory action to the propagation of Lymphoma Raji Cells, after paclitaxel (4 μ g/ml) acts on 48 hours, inhibitory rate of cell growth reaches 82.6%, and through morphology and flow cytometry analysis showed, cell is G 2/ M the phase blocks and apoptosis.
Garlicin, along with garlicin concentration and the increase of action time, human leukemia cell line K 562s phase cell content obviously decline, G 2/ M phase cell content has obvious ascendant trend, prompting garlicin effect be by by cell detention in G 2the result of/M phase.
Indigo Naturalis, indirubin, Meisoindigotin, Indigo Naturalis is one of effective ingredient screening discovery one by one on the DANGGUI LUHUI WAN treatment effective basis of CML.Indirubin is isolated effective ingredient from Indigo Naturalis, can obtain through chemosynthesis.Indirubin has inhibitory action in various degree to untreated slow grain immature granulocyte propagation, and its action principle may be suppress it to breed by suppressing leukaemia DNA synthesis.Meisoindigotin is the similar product of synthesis indirubin, and curative effect is higher than indirubin.Nearest research shows, the mechanism of Meisoindigotin treatment CML may be suppressed relevant with apoptosis with Meisoindigotin leukemogenesis cell proliferation.
Arsenicum, Compendium of Material Medica is early on the books: " arsenic is the medicine of the large poison of large heat, and the poison of arsenicum is especially strong ".Arsenicum sablimatum becomes arsenicum through distillation, and the main chemical compositions of arsenicum is arsenic trioxide or arsenious acid.Arsenic trioxide can cell death inducing, and the expression reducing bcl-2 gene in cell may be one of molecular mechanism of its effect.
Radix Puerariae, find the research of kudzu vine root S86019, S86019 can suppress HL-60 cell proliferation, makes cell be the cell that trend is ripe by promyelocyte phasic development, and the increase acted on along with drug level also becomes to strengthening.S86019 is an effective HL-60 cell induction differentiation agent really.
Elemene, the elemene extracted from Rhizoma Curcumae has obvious impact to Primary human megakaryocytic leukemia cell series Himeg cell cycle, and its effect link mainly affects the cell cycle S phase to G 2the transition process of/M phase, is arrested in the S phase by leukaemia, reduce and enter G 2the cell quantity of/M phase, suppresses leukemic cell growth metabolism, reduces its mitosis, and cause affected tumor cell rapid apoptosis.
Fructus Crotonis, someone is broken up and cytotoxic screening experiment HL-60 cell induction by 302 kinds of Chinese medicines, tentative confirmation Fructus Crotonis has more significant inducing leukemia cell and thinks the effect that normal cell breaks up, and in this result prompting Fructus Crotonis, some composition is expected to become the leukemic active drug for the treatment of.
Radix Sophorae Flavescentis, Radix Sophorae Flavescentis has good therapeutical effect to older patients with acute non-lymphocytic leukemia, and its mechanism may be: to the direct killing effect of tumor cell; Directly act on rake DNA and play its antitumor action; Its antitumor action is played by affecting cell cycle; Its antitumor action is played by inducing apoptosis of tumour cell; Synergism is played by induced microparticle cytochrome P450; By the anti-tumor activity etc. of immunologic mechanism enhancing body.
Qinghuang Powder, Qinghuang Powder [Indigo Naturalis, Realgar (As 2s 3)].Have good curative effect with Qinghuang Powder for Treatment CML, after medication, symptom is clearly better or extinction time is 11.4 days, and within average 10.1 days, spleen starts to reduce.Experimentation shows, Qinghuang Powder is to L 615, S 180dNA, RNA synthesis of cell has inhibitory action in various degree, and be characterized in that inhibitory action is fast, namely drug effect reaches peak in 30 minutes in cell; Drug effect amount effect relationship; Obvious inhibitory action is all had to DNA, RNA.Over the course for the treatment of, the experimental technique such as hematopoietic stem cell cultivation is utilized to observe the impact on mice pluripotent stem cell, grain system and the various cell such as CFU-E, tip hemogram of Qinghuang Powder and Busulfan, result shows that Busulfan has obvious inhibitory action to above-mentioned various types of cells, Qinghuang Powder then unrestraint effect, prompting Qinghuang Powder may have selective inhibitory to leukaemia.Carry out ultrastructure to CML Bone Marrow of Patients cell dynamically to observe, the degeneration necrosis that it is feature that discovery leukaemia mainly occurs with karyolysis, karyopycnosis, fragmentation and Combination, show that the curative effect that Qinghuang Powder for Treatment CML produces is relevant with the pharmacological action of its promotion leukaemia degeneration necrosis, namely finally cause leukaemia dead by degeneration.
No. 1, cancer spirit, cancer No. 1, spirit (pyrrole stone, Calomelas) is current China traditional Chinese medical herbal treatment leukemia particularly one of the most effective medicine of promyelocytic leukemia.Its pharmacology principle is the treating the poisonous disease with poisonous drugs effect of the traditional Chinese medical science.Arsenical is traditional poison in Chinese medicine, and Xin Wen is very toxic.Experimentation proves, arsenic compound can combine with the sulfydryl (-SH) in tissue (protein), protein is lost activity, thus inhibits Leukemia Cell Proliferation, serves the leukemic effect for the treatment of.Secondly, to the selective cytotoxicity of cancerous cell.Experimental studies have found that No. 1, cancer spirit is selective between cancerous cell and normal cell, even if long-term prescription is to hemopoietic function of bone marrow also unrestraint, on the contrary hemopoietic function.3rd, there is induction of differentiation.Use arsenic trioxide injection treatment APL, hemogram and the bone marrow smear of observing patient CR are that after the patient treatment of discovery 77.3%, leukocyte rises gradually, peak is reached in 7-20 days, while leukocyte raises, point apoplexy due to endogenous wind promyelocyte ratio of discovery reduces, and in, metamyelocyte can reach 0.12 and 0.15 respectively.4th, have and promote apoptotic effect.Use during arsenic agent treatment leukemia and find, can see granulocyte degenerative change in patient's bone marrow after treatment CR, endochylema and core have vacuolar degeneration, and chromatin pyknosis, coagulation inequality and nucleorhexis-" apoptotic body ", detect visible Apoptotic cell peak with drain cell instrument.
LIUSHEN WAN, LIUSHEN WAN (Margarita powder, Calculus Bovis, Moschus, Venenum Bufonis, Borneolum Syntheticum) has the merit of heat-clearing and toxic substances removing, blood-activating analgetic.Pharmacology proves that the Calculus Bovis in LIUSHEN WAN contains bilirubin, ergosterol, has effects such as promoting erythrocyte new life, calmness, convulsion, heart tonifying; Venenum Bufonis containing Venenum Bufonis dienol, epinephrine, lysergol, catecholamines, has heart tonifying, boosting, the effect such as anticancer; Realgar is containing orpiment, and sulfydryl enzyme system can be suppressed to affect cellular metabolism, thus have inhibitory action to the growth of tumor cell, other still have Margarita, Borneolum Syntheticum, Moschus etc. to have the effects such as antifungal, anti-bacteria, stimulating central nervous system system.LIUSHEN WAN treatment leukemia, for inducer remission, has obvious inhibitory action to granulocytic systen, but then very unobvious to bone marrow depression, and can prevention and therapy oral ulcer, tonsillitis etc.
FUFANG QINGDAI PIAN, FUFANG QINGDAI PIAN (Indigo Naturalis, Radix Pseudostellariae, Radix Salviae Miltiorrhizae, Realgar etc.) experimentation shows, Realgar has induction APL cell (NB4) and the apoptotic effect of HL-60.
The multiple health of blood, multiple health (Indigo Naturalis, Rhizoma Curcumae, the Radix Puerariae etc.) experimentation of blood confirms, the multiple health of blood can suppress K 562growth of Cells also makes it that apoptosis occur.
Morin (morin): No. CAS: 480-16-0, molecular formula: C 15h 10o 7, chemical name: 3,5,7,2 ', 4 '-pentahydroxyflavone, (3,5,7,2 ', 4 '-Tricetin).Antibacterial, anticancer, inhibitory enzyme activity, there is antioxidation, anti-pain, antibacterial, antiinflammatory, atherosclerosis, the effect such as reduction blood glucose and anti-stress.
Podophyllotoxin (podophyllotoxin): CAS 518-28-5, molecular formula C 22h 22o 8: molecular weight 414.41, physicochemical property: white, needle-shaped crystals powder.Be soluble in chloroform, acetone, ethyl acetate and benzene, dissolve in ethanol, ether, water insoluble.Podophyllotoxin is from XIAONIE section may apple---the wooden lipid antitumor composition that extracts the root of Chinese common sinopodophyllum rhi-zome (also known as chicken tongue element) and stem.For white crystalline powder, odorless, there is hygroscopicity.Chinese common sinopodophyllum rhi-zome is mainly distributed in the ground such as the Gansu Province of China.The podophyllotoxin extracted from Chinese common sinopodophyllum rhi-zome (also known as chicken tongue element) can make unguentum and tincture.Podophyllotoxin effectively can suppress herpesvirus, and the mitosis in T suppression cell mid-term, for toxicity sexually transmitted disease (STD).
Sarcostin (sarcostin): CAS 18607-76-6, molecular formula C 21h 34o 6, molecular weight 382.50.
Megacarpidine (soladulcidine): also known as soladulcine.CAS 511-98-8, molecular formula C 27h 45nO 2, molecular weight 415.66, fusing point 209 ~ 211 DEG C.Optical rotation [α] d-50 ° (c=0.4, chloroform).Naturally to be present in the plants such as the fruit of plant of Solanaceae Fructus Solani Dulcamarae (Herba Solani Lyrati, bittersweet) (Solanum dulcamara L.).There is antifungic action.Inhibit activities is had to Clavicipitaceae, Pyricularia oryzae etc.
4 kinds of medicines structures of pharmaceutical composition of the present invention are as follows:
Morin (morin) podophyllotoxin (podophyllotoxin)
Sarcostin (sarcostin) megacarpidine (soladulcidine).
Summary of the invention
The object of the invention is the deficiency overcoming background technology, provide one to treat leukemic pharmaceutical composition.
The present invention is achieved through the following technical solutions:
Of the present invention a kind of treat the crude drug of leukemic pharmaceutical composition composition and weight portion be:
Morin 10-30 weight portion podophyllotoxin 5-25 weight portion sarcostin 15-20 weight portion megacarpidine 2-18 weight portion.
Of the present inventionly a kind ofly treat leukemic pharmaceutical composition the conventional method of galenic pharmacy can be adopted to be prepared into tablet, capsule, drop pill.
One of the present invention treats leukemic pharmaceutical composition, it is characterized in that being used for the treatment of leukemia.
Gained pharmaceutical composition of the present invention, by regulatory T-cell subset proportions and promote that the secretion of these cytokines plays its immunoregulation effect, recover self suppressed immunologic function and kill and wound residual leukaemia, thus confirm that it has and induce IL-2, the effect of IL-6 and IFN-γ, has certain effect to leukemic therapy apparatus immunity function restructuring.Our experimental result also shows, gained pharmaceutical composition of the present invention has enhancing IL-2 and NCK active function, illustrates that pharmaceutical composition of the present invention has certain protective effect to the immunosuppressant of P388 leukemia mouse after chemotherapy.Infer its mechanism of action may with enhancing Exploration of Red Cell Immuno-adhesive Function, impel erythrocyte to discharge NCK activity factor, regulate NCK-IFN-IL-2 immunological network relevant.
Detailed description of the invention
Treat leukemia medicament composition and method of making the same below by specific experiment example and embodiment to one of the present invention to be described further, but be not limited to the present invention.
Embodiment 1: treat leukemic pharmaceutical composition
Composition and the weight portion for the treatment of the crude drug of leukemic pharmaceutical composition are:
Morin 10 weight portion podophyllotoxin 25 weight portion sarcostin 15 weight portion megacarpidine 18 weight portion
Embodiment 2: treat leukemic pharmaceutical composition
Composition and the weight portion for the treatment of the crude drug of leukemic pharmaceutical composition are:
Morin 30 weight portion podophyllotoxin 5 weight portion sarcostin 20 weight portion megacarpidine 2 weight portion
Embodiment 3: treat leukemic pharmaceutical composition
Composition and the weight portion for the treatment of the crude drug of leukemic pharmaceutical composition are:
Morin 15 weight portion podophyllotoxin 20 weight portion sarcostin 17 weight portion megacarpidine 18 weight portion
Embodiment 4: treat leukemic pharmaceutical composition
Composition and the weight portion for the treatment of the crude drug of leukemic pharmaceutical composition are:
Morin 18 weight portion podophyllotoxin 15 weight portion sarcostin 18 weight portion megacarpidine 10 weight portion
Embodiment 5: the preparation of tablet
Example 1 compositions 150g, adds starch 75g, mixing, granulates, dry, adds microcrystalline Cellulose 20g, magnesium stearate 2.5g, and mixing, is pressed into 1000, obtains present composition tablet.
Embodiment 6: the preparation of capsule
Example 2 compositions 165g, adds starch 65g, mixing, granulates, and dry, granulate, adds appropriate magnesium stearate, and mixing, obtains present composition capsule by encapsulated 1000.
Embodiment 7: the preparation of drop pill
Taking polyethylene glycol 6000 200g water-bath (80 DEG C) heating boils molten, add embodiment 3 compositions 50g, stirring, is coolant with liquid paraffin, puts in glass tubing (4*80cm), chilling temperature is 10 DEG C, drip internal-and external diameter is 7.0/2.0 (mm/mm), and drip is 2cm apart from liquid level, drips speed with per minute 50 for optimum condition, blot the condensing agent on drop pill surface with cotton, obtain present composition drop pill.
Experimental example 1: pharmaceutical composition is to L 615the test of pesticide effectiveness of leukemia mouse
1. modeling method: aseptically get L 615leukemia mouse spleen makes 25% normal saline cell suspension, gives the L that grows up 615mouse hypodermic inoculation, all there is leukemia in result, transplants continuously, set up stable L with morbidity mice spleen oncocyte at Syngenic mice 615leukemia Model.
L 615the foundation of leukemia cell line, removes L 615the spleen of leukemia mouse, shreds spleen in Hanks liquid, makes leukaemia free out, by collected by centrifugation free cell, then uses RPMI1640 culture fluid (containing 10% calf serum) to be diluted to 8*10 6the Cell sap of/ml, cultivates at 37 DEG C.Cultivate 20d and find there is adherent fusiform cells in culture, around it, have the round cell island of propagation.When separately being cultivated by cells different for two kinds of forms, spindle cell can normal adherent growth, and suspension cell growth is slow; Again by two kinds of cells Mixed culture again, two kinds of Growth of Cells are vigorous, after 45d, by the round cell of suspension growth single culture again, pass to for 12 generations always, and Growth of Cells is good, thus sets up this cell strain.
2. the medicine composite for curing leukemia test of pesticide effectiveness.
Material: Kunming mouse (Shandong Province's Experimental Animal Center).Embodiment 1, embodiment 2, embodiment 4 obtain pharmaceutical composition, cyclophosphamide (Hengrui Medicine Co., Ltd., Jiangsu Prov.), standard RhIL-2 (Beijing Shuanglu Pharmaceutical Co., Ltd.), school of life and health sciences cellular resources center, Yackl and IL-2 dependent cells strain CTLL(Chinese Academy of Sciences Shanghai), RPMII640 cell culture fluid and hyclone (Huamei Bio-Engrg Co.); Tetramethyl azo azoles salt (MTT), dodecyl sodium sulfate and canavaline, dimethyl formamide is (Sigma Products); Sample injector, CO 2incubator, ELX-800 microplate reader (Bio-Tek company of the U.S.); The 96 flat Tissue Culture Plates in hole, Epics flow cytometer (Beckman company of the U.S.).
Method: modeling as stated above.Experiment grouping first group is Normal group, and 20 615 is mice, after conventional raising 1w, and intraperitoneal injection of saline 0.2ml/d, normal saline enema 0.3ml, each 1 time of upper and lower noon.2nd group is model group, after conventional raising 1w, and inoculation L 615leukaemia, the same Normal group of inoculation post processing.3rd group is Western medicine group, and 20 conventional raisings, after one week, inoculate L 615leukaemia, after inoculation the 3rd day starts ip cyclophosphamide, 200mg/kg, 1 time/d.4th group of-6 groups is respectively pharmaceutical composition group (embodiment 1), pharmaceutical composition group (embodiment 2), pharmaceutical composition group (embodiment 4), get respectively 20 conventional raise 1d after, inoculation L 615leukaemia, gives pharmaceutical composition group (embodiment 1), pharmaceutical composition group (embodiment 2), pharmaceutical composition group (embodiment 4) gavage on the 3rd day, 0.3ml, each 1 time of upper and lower noon after inoculation respectively.Above each treated animal often group get 10 in the 5th day put to death, get spleen and carry out related immune Indexs measure, remaining each animal carries out observation life cycle.
Observation index: 1. observe life cycle.2. T cell Subsets.3. IL-2, IL-6, IFN-gamma activity measures.
Statistical method: data represent with X ± s, adopts Mi-crosoft Excel to carry out t inspection.
Result:
Life cycle: pharmaceutical composition group of the present invention life cycle, comparatively model group and Western medicine group obviously extended, statistics there were significant differences P<0.01, in table 1.
Table 1 each experimental group compares life cycle
Group Number of elements Life span (d)
Model group 10 5.89±0.45
Western medicine group 10 16.86±1.27 *
Pharmaceutical composition group (embodiment 1) 10 27.77±2.46 *△
Pharmaceutical composition group (embodiment 2) 10 26.45±2.87 *△
Pharmaceutical composition group (embodiment 4) 10 26.55±2.35 *△
Note: compared with model group *p<0.01, compared with Western medicine group △p<0.01.
IL-2, IL-6, IFN-gamma activity: after Western medicine group leukemia mouse chemotherapy, it secretes IL-2, IL-6, the level of IFN-γ significantly declines, and the secretion level of pharmaceutical composition group IL-2 of the present invention, IL-6, IFN-γ obviously increases, both have significant difference (P<0.01), in table 2 at contrast.
Table 2 respectively organizes IL-2, and IL-6, IFN-gamma activity changes
Group Number of elements IL-2 IL-6 IFN-γ
Normal group 10 21.38±1.14 44.46±4.51 12.34±1.87
Model group 10 7.88±1.23 * 16.45±4.32 * 8.87±2.67 *
Western medicine group 10 6.25±1.34 △ 11.27±4.11 △ 5.65±1.76 △
Pharmaceutical composition group (embodiment 1) 10 16.34±2.02 ▲ 32.54±1.67 ▲△ 10.02±1.34 ▲△
Pharmaceutical composition group (embodiment 2) 10 17.43±2.12 ▲ 34.56±2.80 ▲△ 10.34±1.66 ▲△
Pharmaceutical composition group (embodiment 4) 10 17.10±1.89 ▲ 33.14±2.07 ▲△ 10.29±1.89 ▲△
Note: compared with Normal group *p<0.01, compared with model group △p<0.01, compared with Western medicine group ▲p<0.01.
The whether normal of immunologic function is one of key factor of tumor development, and immunologic function is by performed by the cytokine secreted by immunocyte, T cell subset proportions imbalance after leukemia mouse chemotherapy, IL-2, IL-6, the levels of cytokine secretion such as IFN-γ decline, and point out the immunologic function of now body to be in inhibitory state.And after the application of the invention medicine composite for curing, IL-2, IL-6, the levels of cytokine secretion such as IFN-γ go up, illustrate that pharmaceutical composition of the present invention is by promoting that the secretion of these cytokines plays its immunoregulation effect, recovers self suppressed immunologic function to kill and wound residual leukaemia, thus proved invention pharmaceutical composition has and induces IL-2, the effect of IL-6 and IFN-γ, has certain effect to leukemic therapy apparatus immunity function restructuring.
Experimental example 2: pharmaceutical composition is to the pharmacodynamics test of P388 leukemia mouse
1. modeling method: get ascitic type P 388leukemia mouse, de-cervical vertebra is put to death, and abdominal part is sterilized, suction ascites, and add normal saline mixing, it is (1-1.5) * 10 that microscope lowers intact cells concentration 7individual/ml, the inoculum density of every Mus 0.2ml(Kunming mouse and dosage) carry out lumbar injection.
2. pharmaceutical composition is to P 388the impact of natural killer cell (NCK), interleukin-2 (IL-2) activity after leukemia mouse chemotherapy.
Material: Kunming kind one-level mice (Shandong Province's Experimental Animal Center), 50, male and female half and half, body weight 18-22g.Pharmaceutical composition prepared by embodiment 4, cyclophosphamide (Hengrui Medicine Co., Ltd., Jiangsu Prov.), standard RhIL-2 (Beijing Shuanglu Pharmaceutical Co., Ltd.), school of life and health sciences cellular resources center, Yackl and IL-2 dependent cells strain CTLL(Chinese Academy of Sciences Shanghai), RPMII640 cell culture fluid and hyclone (Huamei Bio-Engrg Co.); Tetramethyl azo azoles salt (MTT), dodecyl sodium sulfate and canavaline, dimethyl formamide is (Sigma Products); Sample injector, CO 2incubator, ELX-800 microplate reader (Bio-Tek company of the U.S.); The 96 flat Tissue Culture Plates in hole, Epics flow cytometer (Beckman company of the U.S.).
Modeling and group technology: get 40 mices, inoculation P 388the strain of Lymphocytic leukemia tumor (inoculation method: get ascitic type P388 leukemia mouse, de-cervical vertebra is put to death, and abdominal part is sterilized, suction ascites, adds normal saline mixing, and it is 1.5*107/ml that microscope lowers intact cells concentration, every mice right oxter injection 0.2ml), be divided into 4 groups at random.Another 10 is normal blank group.
Processing method: 1. normal blank group: with 0.4ml/20g normal saline ig; 2. P 388leukemia Model group: with 0.4ml/20g normal saline ig; 3. chemotherapy group: with 0.4ml/20g normal saline ig; 4. chemotherapy adds pharmaceutical composition low dose group: respectively with 50mg pharmaceutical composition in 0.4ml normal saline ig; 5. chemotherapy adds pharmaceutical composition high dose group: be dissolved in 0.4ml normal saline ig with 100mg extract.3. the continuous ig14d of each group above, give simultaneously, 4., 5. organize ip cyclophosphamide 35mg/kg/d, be used in conjunction 6d.In the 15th day, de-cervical vertebra was put to death, and aseptically got spleen NCK, IL-2 active.
Detection method: NCK Activity determination is with reference to Jiang Zhonghua method, and IL-2 activity test method is with reference to Qian Yukun method.
Statistical method: adopt many group means to compare, namely first do variance analysis, remake q inspection.
Result: result shows, it is active that pharmaceutical composition of the present invention can improve NCK and IL-2.In table 3
Table 3 is group NCK and IL-2 expression activitiy respectively
Group n NCK activity (%) IL-2 activity (Iu/ml)
Normal group 8 42.10±2.45 18.66±2.56
Model control group 8 34.34±2.34 * 14.34±2.08 *
Chemotherapy group 8 26.07±2.25 **△△ 10.22±2.38 **△△
Chemotherapy adds pharmaceutical composition low dose group 8 35.33±2.36 ** 14.05±2.56 **
Chemotherapy adds pharmaceutical composition high dose group 8 38.33±2.57 ** 16.76±2.55 ##
Note: compare with normal group, *p<0.05, *p<0.01; Compare with model group, △p<0.05, △ △p<0.01; Compare with chemotherapy group, #p<0.05, ##p<0.01.
NCK is one of three major types lymphocyte, be the important composition composition of the early stage antineoplastic immune supervisory role of body, be a group broad-spectrum anti-tumor cell, its killing activity does not need the prior sensitization of antigen, do not rely on antibody and thymus, not by the restriction of major histocompatibility antigen (MHC) yet.NCK does not need specific antigen to stimulate for virus infected cell and tumor cell in vivo, immune complex and target cell surface structure all directly can bring out the immunoreation of NCK, the a large amount of cytokine of rapid secretion, as interferon-γ (IFN-γ), tumor necrosis factor-alpha (TNF-α), GM-CSF (GM CSF), IL-2 etc., regulates phagocyte and T, B cell function.IL-2, IFN can increase again the activity of NCK simultaneously, therefore body exists NCK-IFN-IL-2 immunological network to have scholar to think.The glycoprotein of the 15KD that IL-2 is mainly produced by T cell, and act on T cell, impel T cell to breed.IL-2 is main, the most strong SCIFs in body.IL-2 can induce or promote the activity of various kinds of cell toxic cell, as NCK, cytotoxic T lymphocyte (CTL), Lymphatic circulation (LAK) and tumor infiltrating lymphocyte (TIL) etc., these cells are most important in the immune surveillance and antineoplastic immune of body.Our experimental result shows, P388 leukemia mouse self and immunity function after chemotherapy are suppressed, NCK and IL-2 activity all has reduction in various degree, and after chemotherapy, P388 leukemia mouse is starkly lower than normal group and model group.This is because chemotherapeutic is on immune impact, also kills and wounds or inhibit caused by LAK cell while killing tumor cell.Recent study finds, the NCK of tumor mice activity reduces, may with body lotus tumor time, the cytokine network antitumor action of its immunocyte is not normal relevant, and as immunosuppressive factor in serum exists and increased activity, immunological enhancement factor active declines.Can by suppressing the generation of IFN thus making NCK activity be suppressed further after giving chemotherapeutic on this basis.In normal body, IL-2 has self forward immunoregulation effect, but at most of tumor mice, IL-2 system exists obviously suppression or disorderly.Most researcher is thought, the ability and all having IL-2 reactivity that tumor patient produces IL-2 reduces in various degree, may SMXO and Ts function strengthen relevant in or host hyperfunction with IL-2 catabolism.After giving chemotherapeutic, inducing of IL-2 is suppressed further, thus increases the weight of the decline of tumor mice immunity.After giving medicine composite for curing of the present invention, high dose group NCK and IL-2 is all increased significantly.Our experimental result shows, Herba Elephantopi Mollis extract has enhancing IL-2 and NCK active function, illustrates that pharmaceutical composition of the present invention has certain protective effect to the immunosuppressant of P388 leukemia mouse after chemotherapy.Infer its mechanism of action may with enhancing Exploration of Red Cell Immuno-adhesive Function, impel erythrocyte to discharge NCK activity factor, regulate NCK-IFN-IL-2 immunological network relevant.

Claims (7)

1. treat a leukemic pharmaceutical composition, it is characterized in that the composition of the crude drug making this pharmaceutical composition and weight portion are:
Morin 10-30 weight portion podophyllotoxin 5-25 weight portion sarcostin 15-20 weight portion megacarpidine 2-18 weight portion.
2. according to claim 1: one treats leukemic pharmaceutical composition, it is characterized in that composition and the weight portion of the crude drug making this pharmaceutical composition are:
Morin 10 weight portion podophyllotoxin 25 weight portion sarcostin 15 weight portion megacarpidine 18 weight portion.
3. according to claim 1: one treats leukemic pharmaceutical composition, it is characterized in that composition and the weight portion of the crude drug making this pharmaceutical composition are:
Morin 30 weight portion podophyllotoxin 5 weight portion sarcostin 20 weight portion megacarpidine 2 weight portion.
4. according to claim 1: one treats leukemic pharmaceutical composition, it is characterized in that composition and the weight portion of the crude drug making this pharmaceutical composition are:
Morin 15 weight portion podophyllotoxin 20 weight portion sarcostin 17 weight portion megacarpidine 18 weight portion.
5. according to claim 1: one treats leukemic pharmaceutical composition, it is characterized in that composition and the weight portion of the crude drug making this pharmaceutical composition are:
Morin 18 weight portion podophyllotoxin 15 weight portion sarcostin 18 weight portion megacarpidine 10 weight portion.
6. one according to claim 1 treats leukemic pharmaceutical composition, and the conventional method of galenic pharmacy can be adopted to be prepared into tablet, capsule, drop pill.
7. one according to claim 1 treats leukemic pharmaceutical composition, it is characterized in that being used for the treatment of leukemia.
CN201410628570.5A 2014-11-11 2014-11-11 Pharmaceutical composition used for treating leukemia Pending CN104473946A (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2016197944A1 (en) * 2015-06-12 2016-12-15 浙江省医学科学院 Uses of antidepressant compound in preparation of antidepressant drugs and antidepressant health foods

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
HSIU-MAAN KUO等: "《Morin Inhibits the Growth of Human Leukemia HL-60 Cells via Cell Cycle Arrest and Induction of Apoptosis through Mitochondria Dependent Pathway》", 《ANTICANCER RESEARCH》 *
孙立新 等: "《中药白英的研究进展》", 《沈阳药科大学学报》 *
陈静等: "《鬼臼毒素纳米脂质体对人白血病细胞的凋亡诱导作用》", 《中国临床药理学与治疗学》 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2016197944A1 (en) * 2015-06-12 2016-12-15 浙江省医学科学院 Uses of antidepressant compound in preparation of antidepressant drugs and antidepressant health foods
US10391104B2 (en) * 2015-06-12 2019-08-27 Zhejiang Academy Of Medical Sciences Application of antidepressant compound in preparation of antidepressant drugs and antidepressant health-care foods

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