CN102357100A - Anti-tumor combination medicament - Google Patents
Anti-tumor combination medicament Download PDFInfo
- Publication number
- CN102357100A CN102357100A CN2011103068750A CN201110306875A CN102357100A CN 102357100 A CN102357100 A CN 102357100A CN 2011103068750 A CN2011103068750 A CN 2011103068750A CN 201110306875 A CN201110306875 A CN 201110306875A CN 102357100 A CN102357100 A CN 102357100A
- Authority
- CN
- China
- Prior art keywords
- copper
- disulfiram
- tumor
- dosage
- combination 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.)
- Pending
Links
Images
Abstract
The invention relates to a medicament in combination of a disulfiram preparation and copper, having an anti-tumor function. The combination of disulfiram and copper in the invention plays a role in treating tumor cells, belonging to the technical field of medicaments. The anti-tumor combination medicament comprises disulfiram or derivatives thereof and copper and any one or more optional medicinal vectors, which are used simultaneously, respectively or sequentially; the dosage of the disulfiram in each single medicament ranges from 2 to 200 mg/kg or equals to the equivalent dosage of disulfiram derivatives; and the copper comprises micro copper ions contained in the human body and other absorbable forms of copper irons, wherein the dosage of the copper in each single medicament is 0.1-5 mg per day. The combination medicament disclosed by the invention has remarkable effect in treating tumors and reduced toxic and side effects.
Description
Technical field
The present invention relates to have the disulfiram preparation of antitumor action and the combination medicine of copper, disulfiram of the present invention and copper are united use tumor cell are had therapeutical effect, belong to medical technical field.
Background technology
(disulfiram DS), has another name called tetraethylthiuram disulfide, alleviating alcohol addiction sulfur to disulfiram, and structure such as Fig. 1 were synthesized out first in 1881, is used as the catalyst of the vulcanization of rubber in the rubber industry at first; The 1830's, DS was for the first time as medicine, as the treatment of scabies; Subsequently, it is found that DS and copper ion have very strong chelation, can reduce the activity of cupric respiratory enzyme, and the enteral anthelmintic is very responsive to DS, so DS is used as the anthelmintic use again; Two the doctor Hald and the Jacobsen in Copenhagen in 1948 chance on, and drink a small amount of wine after taking the anthelmintic disulfiram, can cause symptoms such as flush, headache, stomachache, perspiration, cardiopalmus, dyspnea immediately.People call disulfiram-like reaction to this symptom that occurs of behind the contact disulfiram, drinking.Just be based on disulfiram-like reaction, DS is using as antialcoholic always since over half a century.Its structural formula is following:
Prior art mainly discloses the preparation and the effect aspect antibiotic, anti-cataract thereof of disulfiram." reparation technology of tetraethylthiuram disulfide " of Superfine Chemical Co Ltd, Leqing City's invention disclosed in Chinese patent publication number CN 1316422A document.It relates to the reparation technology of having invented a kind of tetraethylthiuram disulfide, but does not relate to its application and preparation research as medicine.
" the antibiotic therapeutic alliance " of the biological company limited invention of e-disclosed in Chinese patent publication number CN 101939026 A documents.It relates to having invented and comprises the imidazoles or derivatives thereof with therapeutic activity and the compositions of disulfiram or derivatives thereof, causes or the combination preparation of the infection that causes as being used to treat by the multi-drug resistant antibacterial.
" a kind of high stable anti-cataract medicine " of Shenyang Pharmaceutical University's invention disclosed in Chinese patent publication number CN 1408418A document.It relates to has invented a kind of (DDC)2Zn (DDC)
2The hydroxypropyl-beta-cyclodextrin inclusion eye drop of Zn is a kind of high stable anti-cataract medicine, has the characteristics of high biological utilisation, high security and high stability.
Shenyang Pharmaceutical University's invention " inclusion compound of disulfiram as eye drops and preparation method thereof " disclosed in Chinese patent publication number CN 1376463A.It relates to has invented inclusion compound eye drops that a kind of disulfiram and HP-process and preparation method thereof, can add proppant and process the lyophilizing eye drop; Have good water solubility, stable strong characteristics, can prevent and treat cataract preferably.
Combinatorx, Inc.'s invention " being used to treat the compositions and the method for tumor " disclosed in Chinese patent publication number CN 101217956A.It relates to the test kit of the method for having invented the multiple compositions that is used to treat the medicine of tumor patient, treatment tumor, the multiple medicine that is used to treat cancer and possibly be used to treat the authentication method of the chemical compound combination of tumor patient; Wherein comprised the combination of alleviating alcohol addiction sulfur (being disulfiram) with the anti-inflammatory agent auranofin.
Zhang Hong etc. have prepared anti-cataract medicine disulfiram Emulsion based on the anti-cataract effect of disulfiram, wherein with Ovum Gallus domesticus Flavus lecithin as emulsifying agent; Adopt ultrasonic method; Investigated the chemical stability of disulfiram in the disulfiram Emulsion, measured different Emulsions and formed influence, drawn the disulfiram degraded and obey first order reaction disulfiram stability; Speed constant and temperature relation meet Arrhenius equation, and wherein better prescription can 4 ℃ be preserved 695 days.
Through document retrieval, do not see that disulfiram and copper are united to use the report of treating the tumor aspect.
Summary of the invention
Inventing technical problem to be solved is; A kind of new antineoplastic combination drug is provided; Adopt the disulfiram preparation or adopt the disulfiram preparation and the Combined application of copper, the disulfiram preparation can the efficacious therapy tumor, and the Combined application of disulfiram preparation and copper is to two kinds of medicines generation synergism; And then the more efficiently treatment tumor of ability, reduce toxic and side effects simultaneously.
In order to solve the problems of the technologies described above, the present invention realizes through following technical proposals.
Antineoplastic combination drug, it comprises the disulfiram or derivatives thereof; The disulfiram or derivatives thereof and the copper of difference or use in order simultaneously.
Described antineoplastic combination drug comprises that the disulfiram or derivatives thereof that gives object treatment effective dose, the copper that gives the treatment target effective dose give the disulfiram or derivatives thereof of treatment target effective dose simultaneously; Wherein, described disulfiram is 2-200mg/kg for each single agent dose scope of adult, or the disulfiram derivant of equivalent; Described copper-clad is drawn together trace copper ion that human body contains and copper ion, and other can absorb form, and wherein copper is 0.1-5mg/ day for single agent dose of adult; Medicine can be tried to achieve through people and the calculating of animal dose,equivalent reduction formula for the dosage of other animals.
The derivant of described disulfiram has following basic structure:
R wherein
1, R
2, R
3And R
4Can respectively do for oneself H and replacement or unsubstituted alkyl, thiazolinyl, alkynyl, aryl, alkoxyl, halogen, pharmaceutically acceptable ester, pharmaceutically acceptable thioesters, pharmaceutically acceptable ether, pharmaceutically acceptable thioether, pharmaceutically acceptable inorganic ester.
Described copper is any one copper-containing compound in copper gluconate, copper sulfate, copper chloride, the copper amino acid chelate.
Described disulfiram and copper are processed independent preparation respectively, and the dosage form of two kinds of preparations is identical or different.In use, can copper and two kinds of medicines of disulfiram successively be used, also can use simultaneously.When two kinds of medicines were successively used, the independent preparation of copper can use earlier.
Described disulfiram becomes single compound preparation with copper, use simultaneously.
The uniting use and can realize of disulfiram and copper: oral administration, drug administration by injection (comprising multiple injecting pathway such as intravenous injection, subcutaneous injection, intradermal injection, intramuscular injection, intracavitary administration), respiratory tract administration, percutaneous drug delivery, mucosa delivery, cavity/canal drug administration etc. through various administering modes.
Various dosage forms realizations are processed in the use of uniting of disulfiram and copper respectively or jointly: powder, tablet, granule, capsule, solution, powder pin, Emulsion, liposome, suspensoid, gel, spray, aerosol, aerosol, powder spray, lotion, liniment, ointment, plaster, paste, patch, suppository, drop or drop pill.
Described tumor is any one in leukemia, lymphoma, skin carcinoma, sarcoma, breast carcinoma, hepatocarcinoma, pulmonary carcinoma, glioma or the glioblastoma.
Disulfiram preparation and copper are united when using, and need the dosage of the disulfiram of generation effect can be according to patient disease character and order of severity appropriate change; The dosage of copper can not surpass 5mg/ day.
Owing to adopt technique scheme to make the present invention have following characteristics:
The applicant through experimental study find disulfiram can targeting in a plurality of signal paths; Have positive role at anti-tumor aspect:
DS can form blended disulphide with crucial sulfydryl effect in type, the II; Thereby the activity that suppresses type and type; Stop untwisting, transcribe and duplicating of tumor cell dna double chain, and cause the fracture of tumor cell double center chain or single stranded DNA;
metalloproteases (MMPs) is the endopeptidase that zinc is contained in the active center, and the transfer of it and tumor cell is closely related; DS through with the zinc chelating in MMPs active center, suppress the activity of MMPs, reduce the expression of tumor cell line MMP-2 and MMP-9, what stop tumor cell invades profit and transfer;
nuclear factor (NF-κ B) is the factor that and anti-apoptotic and chemotherapy resistance property have substantial connection; DS is through the active expression that suppresses NF-κ B indirectly of CKIs enzyme body; And then existence, propagation and the transfer of inhibition tumor cell; Reduced the expression of the ATP efflux pump of P-glycoprotein (P-gp), many drug resistances associated protein 1 (MRP1) and mitoxantrone drug resistance GAP-associated protein GAP simultaneously; Reduce the chemotherapeutics drug resistance, increase the sensitivity of drug-resistant tumor cell chemotherapeutics;
DS can combine with copper and the zinc in active oxygen (ROS) avtive spot; Suppress ultra oxygen compound dismutase (SOD) activity; Increase ROS content in the cell, inducing apoptosis of tumour cell.The more important thing is that people's such as Daniel the cytotoxicity of discovering DS has the tumor cell specificity, this is relevant with the Cu (II) of high concentration in the tumor cell.DS more be prone to tumor cell in Cu (II) form copper ion complex (Cu (DDC)
2), the inhibitory action that DS goes the same way to proteasome NF-κ B is exactly through Cu (DDC)
2Realize, and the Cu (DDC) that produces in the normal cell
2Measure the less activity that is not enough to CKIs enzyme body.Disulfiram and copper have produced synergism, and then can more efficiently treatment tumor.The present invention is intended to disulfiram is prepared into multiple dosage form, like tablet, capsule, solution, Emulsion etc., and through different modes of administration, as: oral, injection, suction etc., unite use with Cu (II), tumor is had therapeutical effect.The disulfiram preparation can the efficacious therapy tumor, and the Combined application of disulfiram preparation and copper produces synergism to two kinds of medicines, and then can more efficiently treatment tumor, reduces toxic and side effects simultaneously.
Description of drawings
Fig. 1 is that the relevant disulfiram of copper is to breast cancer cell toxicity MCF7 result of the test figure.
Fig. 2 is that the relevant disulfiram of copper is to breast cancer cell toxicity MDA-MB-231 result of the test figure.
Fig. 3 is that the relevant disulfiram of copper is to breast cancer cell toxicity T 47D result of the test figure.
Fig. 4 is the cytotoxicity MCF7 result of the test figure of disulfiram Emulsion and copper coupling.
Fig. 5 is the cytotoxicity MDA-MB-231 result of the test figure of disulfiram Emulsion and copper coupling.
Fig. 6 is the cytotoxic T 47D result of the test figure of disulfiram Emulsion and copper coupling.
Fig. 7 is that DS solution, DS Emulsion and copper are united use for two kinds of cell line cytotoxicities of GBM result.
Fig. 8 is the result of the test figure to the S180 sarcomata that DS solution, DS Emulsion and copper are united use.
Fig. 9 is that DS solution, DS Emulsion and copper are united use for rat body weight situation of change experimental result picture.
The specific embodiment
Below in conjunction with embodiment the present invention is described further.Following examples are merely several specific embodiment of the present invention, but design concept of the present invention is not limited thereto, and allly utilize this design that the present invention is carried out the change of unsubstantiality, all should belong to the behavior of invading protection domain of the present invention.
Method among the following embodiment if no special instructions, is conventional method.
Percentage composition among the following embodiment is the quality percentage composition if no special instructions.
Disulfiram and copper are united the cytotoxicity of use to breast carcinoma:
Investigated disulfiram to different breast cancer cell lines---the cytotoxicity of MCF7, MDA-MB-231 and T47D.
1. material and method
1.1 tumor cell line and cell culture
Breast cancer cell line, MCF7, MDA-MB-231 and T47D are available from LGC standard (ATCC), Britain.Rich copper (Cu) cell is through μ Μ CuCl every days at least 5
2Cultured cell obtains for three days on end.These cell lines are cultivated in the DMEM culture medium, add 10% hyclone, 1% (v/v) penicillin-streptomycin (10000U/mL) and 1% (v/v) L-glutamic acid (200mM).
1.2 the direct cytotoxicity test experiments of mononuclear cell (MTT) step
Test the cell of required cultivation in containing the culture fluid of serum, the tiling density of cell is 5000 cells/well in 96 orifice plates.After the overnight incubation, culture medium replaces to and contains disulfiram and/or copper chloride (CuCl
2) the blood plasma culture fluid, continue to cultivate 72h.Test the cell of required cultivation in serum-free medium, the tiling density of cell is 20000 cells/well in 96 orifice plates.After the overnight incubation, clean cell, then cell culture is being contained 1% (v/v) penicillin-streptomycin (10000U/mL) and 1% (v/v) L-glutamic acid (200mM), disulfiram and/or copper chloride (CuCl with warm in advance phosphate buffer (PBS)
2) serum-free DMEM culture medium in, cultivate 72h.Through after 72 hours hatch, add 50 μ LMTT solution to every hole, hatch 4h with the masking foil parcel.Afterwards, remove culture medium, add 175 μ LDMSO and 25 μ L glycine buffers, obtain the absorbance of 540nm.
2. experimental result
Shown in Fig. 1-3, disulfiram to cultivate three kinds of breast cancer cell lines in the blood plasma medium (+S+DS-Cu) cytotoxicity is arranged all, MCF and MDA-MB-231 have two-way function to the cytotoxicity of disulfiram; The T47D cell is not observed this two-way function.MCF7, MDA-MB-231 and T47D no difference of science of statistics.When in cell culture fluid, adding 1 μ Μ copper chloride (CuCl
2) (+S+DS+Cu) time, the cytotoxicity of DS strengthens, and statistical result is (p＜0.05) significantly.But, when not containing blood plasma in the culture fluid of breast cancer cell (S+DS-Cu) time, even disulfiram content does not show the cytotoxicity to MCF7 and MDA-MB-231 up to 40 μ Μ yet; Yet,, T47D is had cytotoxicity (p＜0.001) when disulfiram concentration when higher (IC50=28.657 μ Μ is higher than 1.215 μ Μ).This shows, at 1 μ Μ copper chloride (CuCl
2) incubation conditions under, copper (II) has booster action to the cytotoxicity of disulfiram; Yet when lacking the blood plasma condition, (S-DS+Cu) toxicity to tumor cell does not almost have in disulfiram and copper (II) associating.
The disulfiram that table 1 copper is correlated with is to the cytotoxicity experiment result of breast carcinoma.
Being IC50 (μ Μ) shown in the table, is SD in the bracket.
*The concentration that IC50 has prepared above disulfiram or CuCl2 in the test is so result＞40 represent IC50 than DS or copper chloride (CuCl in this test
2) maximal dose also high.
Disulfiram and copper coupling are to the Cytotoxic influence of other antitumor drug experiment:
Disulfiram has high cytotoxicity to the tumor of general types, comprise glioblastoma multiforme (glioblastoma multiforme, GBM).Disulfiram can strengthen the cytotoxicity of medicine to tumor cell equally, the normal cell of protection kidney, gastrointestinal tract and bone marrow, thereby improve the anticancer therapy index, for example gemcitabine (gemcitabine, dFdC), cyclophosphamide, cisplatin and ionizing radiation.
The combination of table 2 different pharmaceutical is to the cytotoxicity experiment result of GBM cell line
| ? | U251MG | U87MG |
| IC50s of DS | ? | ? |
| DS alone | ＞10000 | ＞10000 |
| DS + Cu 1μΜ | 143.3(0.8) | 202.3(6.1) |
| IC50s of dFdC | ? | ? |
| dFdC alone | 100.4(22.3) | ＞400 |
| dFdC : DS/Cu1μΜ | 17.5(5.1) | 17.1(3.0) |
| CI | ? | ? |
| IC50 | 0.39 | 0.47 |
| IC75 | 0.33 | 0.46 |
| IC90 | 0.31 | 0.45 |
From table, can know that use disulfiram less to the toxicity of tumor cell separately, 503nhibiting concentration (IC50s) is all greater than 10000; And when share with 1 μ Μ copper, be 143.3 and 202.3 to the IC50s of U251MG and U87MG, cytotoxicity strengthens.IC50s when gemcitabine and disulfiram (mol ratio 10:1), 1 μ Μ copper (II) are united use than two kinds of cells of the independent use processing of gemcitabine is reduced to below 1/5th, and cytotoxicity significantly increases.Association index (CI, combination index) has reflected the effect situation of different pharmaceutical Combined application, and CI＜1 shows to have synergism; CI is more little, explains that synergism is strong more, and CI＜0.5 shows to have very strong synergism.Disulfiram, copper and gemcitabine are united use, and the CI value explains that all less than 0.5 disulfiram, copper and gemcitabine have very strong synergism.The result shows that disulfiram and copper coupling can strengthen the toxicity of disulfiram to tumor cell itself, strengthen the toxicity of other medicines to tumor cell simultaneously.
Embodiment 3
The tumor suppression experiment of low dosage disulfiram:
1. material and method:
Tumor inoculation: adopt the BALB/c-nu male nude mouse to experimentize.Every group three lotus people hepatocarcinoma HepG2 carcinoma nude mices, routine disinfection nude mice skin of chest abdomen; Under aseptic, peel off the tumor piece, remove tumor piece both central necrotic tissue and tumor piece fiber and fatty tissue on every side, tumor tissues is cut into the tumor tissue piece of 1.5 * 1.5 * 1.5mm3 size.The little otch of nude mice right fore armpit skin, it is subcutaneous that tumor tissues is implanted nude mice right fore axillary fossa.
Experimental procedure: after transplanting the strain of people's hepatocarcinoma HepG2 carcinoma, carried out tumor bearing nude mice in 8-10 days and weigh, and with vernier caliper measurement tumor major diameter (a) and minor axis (b), by formula V=a * b
2/ 2, be converted into gross tumor volume.Select the close mice of gross tumor volume and divide into groups, every group of 7 mices.Every 2d is with a, the b of vernier caliper measurement tumor after the administration, and 8d puts to death mice after the administration, separates and cuts the tumor tissues weighing, calculates tumour inhibiting rate (TI).TI (%)=(matched group average tumor quality-administration group average tumor quality)/matched group average tumor quality * 100%
2. dosage regimen:
Administration group tail vein injection disulfiram solution (1mg/ml), dosage 10mg/kg; The matched group tail vein injection saline.
3. experimental result:
The administration group is 38.68% to the tumour inhibiting rate of lotus people hepatocarcinoma HepG2 carcinoma mice, compares with matched group that there were significant differences (P＞0.05).
The described injection disulfiram of present embodiment solution can be substituted by other preparation of medicine effective quantity.Example emulsion, liposome, suspensoid, gel, spray, aerosol, aerosol, powder spray, lotion, liniment, ointment, plaster, paste, patch, suppository, drop or drop pill.
Embodiment 4
The tumor suppression experiment of middle dosage disulfiram:
1. material and method are with embodiment 3;
2. experimental procedure: the strain of hepatocarcinoma carcinoma is by pulmonary carcinoma A549 tumor strain replacement, and other is with embodiment 3;
3. dosage regimen:
Administration group tail vein injection disulfiram Emulsion (10mg/ml), dosage 30mg/kg; The blank Emulsion of matched group tail vein injection.
4. experimental result:
The administration group is 49.32% to the tumour inhibiting rate of lotus people pulmonary carcinoma A549 carcinoma mice, compares with matched group that there were significant differences (P＞0.05).
The tumor suppression experiment of high dose disulfiram:
1. material and method are with embodiment 3;
2. experimental procedure: the strain of hepatocarcinoma carcinoma is by pulmonary carcinoma A549 tumor strain replacement, and other is with embodiment 3;
3. dosage regimen:
Administration group tail vein injection disulfiram Emulsion (10mg/ml), dosage 100mg/kg; The blank Emulsion of matched group tail vein injection.
4. experimental result:
The administration group is 53.77% to the tumour inhibiting rate of lotus people pulmonary carcinoma A549 carcinoma mice, compares with matched group that there were significant differences (P＞0.05).
The described disulfiram Emulsion of present embodiment can be substituted by other external preparation of medicine effective quantity.For example solution, liposome, suspensoid, gel, spray, aerosol, aerosol, powder spray, lotion, liniment, ointment, plaster, paste, patch, suppository, drop or drop pill.
Embodiment 6
Low dosage disulfiram and copper are united the tumor suppression test of use
1. material and method:
Tumor inoculation: adopt the BALB/c-nu male nude mouse to experimentize.Every group three lotus application on human skin cancer A431 cell line carcinoma nude mices, routine disinfection nude mice skin of chest abdomen; Under aseptic, peel off the tumor piece, remove tumor piece both central necrotic tissue and tumor piece fiber and fatty tissue on every side, tumor tissues is cut into the tumor tissue piece of 1.5 * 1.5 * 1.5mm3 size.The little otch of nude mice right fore armpit skin, it is subcutaneous that tumor tissues is implanted nude mice right fore axillary fossa.
2. experimental procedure: after transplanting the strain of application on human skin cancer A431 tumor, carried out tumor bearing nude mice in 8-10 days and weigh, and with vernier caliper measurement tumor major diameter (a) and minor axis (b), by formula V=a * b
2/ 2, be converted into gross tumor volume.Select the close mice of gross tumor volume and divide into groups, every group of 7 mices.Every 2d is with a, the b of vernier caliper measurement tumor after the administration, and 8d puts to death mice after the administration, separates and cuts the tumor tissues weighing, calculates tumour inhibiting rate (TI).TI (%)=(matched group average tumor quality-administration group average tumor quality)/matched group average tumor quality * 100%
3. dosage regimen:
Before the administration for three days on end every day oral glucose aldehydic acid copper solution, dosage 30 μ g/ only/day, the dosage that is equivalent to copper be 4.3ug/ only/day (calculate through people Mus Equivalent Conversion, being equivalent to body weight is copper dosage 1.4mg/ people/day of 60kg adult).
The 4th day, administration group tail vein injection disulfiram solution (1mg/ml), dosage 10mg/kg; The matched group tail vein injection saline.
4. experimental result:
The administration group is 48.17% to the tumour inhibiting rate of lotus application on human skin cancer A431 cell line carcinoma mice, compares with matched group that there were significant differences (P＞0.05).
Embodiment 7
Middle dosage disulfiram and copper are united the tumor suppression test of use
1. material and method are with embodiment 6;
2. experimental procedure: the strain of skin carcinoma A431 tumor is by the replacement of lymphoma Hut-102 cell line, and other is with embodiment 6;
3. dosage regimen:
Before the administration for three days on end every day oral copper-bath, dosage 30 μ g/ only/day, the dosage that is equivalent to copper be 12ug/ only/day (calculate through people Mus Equivalent Conversion, being equivalent to body weight is copper dosage 4mg/ people/day of 60kg adult).The 4th day, administration group tail vein injection disulfiram Emulsion (10mg/ml), dosage 30mg/kg; The blank Emulsion of matched group tail vein injection.
4. experimental result:
The administration group is 57.78% to the tumour inhibiting rate of lotus lymphoma Hut-102 carcinoma mice, compares with matched group that there were significant differences (P＞0.05).
Embodiment 8
High dose disulfiram and copper are united the tumor suppression test of use
1. material and method are with embodiment 6;
2. experimental procedure: the strain of skin carcinoma A431 tumor is by sarcoma cell S-180 tumor strain replacement, and other is with embodiment 6;
3. dosage regimen:
Before the administration for three days on end every day the tail vein injection copper chloride solution, dosage 0.63 μ g/ only, the dosage that is equivalent to copper be 0.3ug/ only/day (calculate through people Mus Equivalent Conversion, being equivalent to body weight is copper dosage 0.1mg/ people/day of 60kg adult).The 4th day, administration group tail vein injection disulfiram Emulsion (10mg/ml), dosage 100mg/kg; The blank Emulsion of matched group tail vein injection.
4. experimental result:
The administration group is 72.61% to the tumour inhibiting rate of lotus sarcoma cell S-180 carcinoma mice, compares with matched group that there were significant differences (P＞0.05).
The used copper chloride solution of present embodiment can be substituted by copper bearing other preparations of equivalent, for example powder, tablet, granule, capsule, solution.
Embodiment 9
The tumor suppression experiment of oral disulfiram and copper compound preparation
1. material and method are with embodiment 6;
2. experimental procedure: the strain of skin carcinoma A431 tumor is by the replacement of leukemia U937 cell line, and other is with embodiment 6;
3. dosage regimen:
The compound granular of preparation copper methionine and disulfiram faces with preceding and disperses the tumor-bearing mice gastric infusion with distilled water.The experimental group tumor-bearing mice, methionine dosage 84 μ g/, the dosage that is equivalent to copper is 15ug//day (calculate through people Mus Equivalent Conversion, being equivalent to body weight is copper dosage 5mg/ people/day of 60kg adult), disulfiram dosage 200mg/kg; Not administration of matched group.
4. experimental result:
The administration group is 57.31% to the tumour inhibiting rate of lotus leukemia U937 carcinoma mice, compares with matched group that there were significant differences (P＞0.05).
The described leukemia of present embodiment is any one in acute leukemia, acute lymphoblastic leukemia, acute myeloid leukemia, acute myeloblastic leukemia, acute promyelocytic leukemia, acute myelomonocytic leukemia, acute monocytic leukemia, Di Guglielmo syndrome, chronic leukemia, chronic granulocytic leukemia, the chronic lymphatic leukemia.
Disulfiram preparation and copper are united use to cytotoxicity
Material and method are with embodiment 1.
Fig. 4-Fig. 6 MTT result show that disulfiram (DS) Emulsion and disulfiram (DS) solution use separately or with the cytotoxicity experiment result of copper coupling to three kinds of cell lines of breast carcinoma.Disulfiram uses as solution separately all has the growth inhibited effect to three kinds of breast cancer cells, and after disulfiram was prepared into Emulsion, this effect was weakened, and tumor cell is to the two-way function enhancing of disulfiram.But, unite and use 1uM copper chloride (CuCl
2), disulfiram Emulsion is to the toxicity of breast cancer cell, and the DMSO solution comparison with disulfiram obviously increases.This shows that the synergism of copper and disulfiram can further strengthen through improving the dosage form of disulfiram.
Fig. 7-Fig. 9 is that disulfiram (DS) solution, disulfiram (DS) Emulsion and copper are united the cytotoxicity result of use.Visible from A figure, between 72 hours culture periods, for two kinds of cell lines of GBM---U251 and U87, DS Emulsion shows higher vitro cytotoxicity than DS solution.Visible from B figure, to S180 sarcomata in the rat body, DS Emulsion associating copper uses and shows the strongest antitumaous effect.DS：32mg/kg,?i.v.;?Cu:?CuCl
2?3.2mg/kg?p.o.。Tumor was in excision in the 21st day.C figure is the rat body weight situation of change.
Embodiment 11
The tumor suppression experiment of dipropyl two thiocarbamate esters:
1. material and method:
Tumor inoculation: adopt the BALB/c-nu male nude mouse to experimentize.Every group three lotus people hepatocarcinoma HepG2 carcinoma nude mices, routine disinfection nude mice skin of chest abdomen; Under aseptic, peel off the tumor piece, remove tumor piece both central necrotic tissue and tumor piece fiber and fatty tissue on every side, tumor tissues is cut into the tumor tissue piece of 1.5 * 1.5 * 1.5mm3 size.The little otch of nude mice right fore armpit skin, it is subcutaneous that tumor tissues is implanted nude mice right fore axillary fossa.
Experimental procedure: after transplanting the strain of people's hepatocarcinoma HepG2 carcinoma, carried out tumor bearing nude mice in 8-10 days and weigh, and with vernier caliper measurement tumor major diameter (a) and minor axis (b), by formula V=a * b
2/ 2, be converted into gross tumor volume.Select the close mice of gross tumor volume and divide into groups, every group of 7 mices.Every 2d is with a, the b of vernier caliper measurement tumor after the administration, and 8d puts to death mice after the administration, separates and cuts the tumor tissues weighing, calculates tumour inhibiting rate (TI).TI (%)=(matched group average tumor quality-administration group average tumor quality)/matched group average tumor quality * 100%
2. dosage regimen:
The administration group is irritated stomach and is given dipropyl two thiocarbamate ester suspensions (5mg/ml), dosage 10mg/kg; Not administration of matched group.
3. experimental result:
The administration group is 27.51% to the tumour inhibiting rate of lotus people hepatocarcinoma HepG2 carcinoma mice, compares with matched group that there were significant differences (P＞0.05).
The tumor suppression experiment of diethyldithio-carbamate methyl ester:
1. material and method:
Tumor inoculation: adopt the BALB/c-nu male nude mouse to experimentize.Every group three lotus people hepatocarcinoma HepG2 carcinoma nude mices, routine disinfection nude mice skin of chest abdomen; Under aseptic, peel off the tumor piece, remove tumor piece both central necrotic tissue and tumor piece fiber and fatty tissue on every side, tumor tissues is cut into the tumor tissue piece of 1.5 * 1.5 * 1.5mm3 size.The little otch of nude mice right fore armpit skin, it is subcutaneous that tumor tissues is implanted nude mice right fore axillary fossa.
Experimental procedure: after transplanting the strain of people's hepatocarcinoma HepG2 carcinoma, carried out tumor bearing nude mice in 8-10 days and weigh, and with vernier caliper measurement tumor major diameter (a) and minor axis (b), by formula V=a * b
2/ 2, be converted into gross tumor volume.Select the close mice of gross tumor volume and divide into groups, every group of 7 mices.Every 2d is with a, the b of vernier caliper measurement tumor after the administration, and 8d puts to death mice after the administration, separates and cuts the tumor tissues weighing, calculates tumour inhibiting rate (TI).TI (%)=(matched group average tumor quality-administration group average tumor quality)/matched group average tumor quality * 100%
2. dosage regimen:
Administration group tail vein injection diethyldithio-carbamate methyl ester injection (1mg/ml), dosage 10mg/ml; Matched group intravenous injection normal saline.
3. experimental result:
The administration group is 39.14% to the tumour inhibiting rate of lotus people hepatocarcinoma HepG2 carcinoma mice, compares with matched group that there were significant differences (P＞0.05).
Claims (9)
1. antineoplastic combination drug is characterized in that it comprises the disulfiram or derivatives thereof; The disulfiram or derivatives thereof and the copper of difference or use in order simultaneously.
2. antineoplastic combination drug according to claim 1 is characterized in that described disulfiram is 2-200mg/kg for each single agent dose scope of adult, or the disulfiram derivant of equivalent; Described copper-clad is drawn together trace copper ion that human body contains and copper ion, and other can absorb form, and wherein copper is 0.1-5mg/ day for single agent dose of adult; Medicine can be tried to achieve through people and the calculating of animal dose,equivalent reduction formula for the dosage of other animals.
3. antineoplastic combination drug according to claim 1 and 2 is characterized in that the derivant of described disulfiram has following basic structure:
R wherein
1, R
2, R
3And R
4Can respectively do for oneself H and replacement or unsubstituted alkyl, thiazolinyl, alkynyl, aryl, alkoxyl, halogen, pharmaceutically acceptable ester, pharmaceutically acceptable thioesters, pharmaceutically acceptable ether, pharmaceutically acceptable thioether, pharmaceutically acceptable inorganic ester.
4. according to antineoplastic combination drug according to claim 1 and 2, it is characterized in that described copper is any one copper-containing compound in glucuronic acid copper, copper sulfate, copper chloride, the copper amino acid chelate.
5. antineoplastic combination drug according to claim 1 and 2; It is characterized in that described disulfiram and copper processes independent preparation respectively, the dosage form of two kinds of preparations is identical or different, in use; Can copper and two kinds of medicines of disulfiram successively be used; Also can use simultaneously, when two kinds of medicines were successively used, the independent preparation of copper can use earlier.
6. antineoplastic combination drug according to claim 1 and 2 is characterized in that described disulfiram becomes single compound preparation with copper, uses simultaneously.
7. antineoplastic combination drug according to claim 1 and 2 is characterized in that uniting use and can realizing through various administering modes of disulfiram and copper: oral administration, drug administration by injection, respiratory tract administration, percutaneous drug delivery, mucosa delivery, cavity/canal drug administration etc.
8. antineoplastic combination drug according to claim 1 and 2 is characterized in that various dosage forms realizations are processed in the use of uniting of disulfiram and copper respectively or jointly: powder, tablet, granule, capsule, solution, powder pin, Emulsion, liposome, suspensoid, gel, spray, aerosol, aerosol, powder spray, lotion, liniment, ointment, plaster, paste, patch, suppository, drop or drop pill.
9. antineoplastic combination drug according to claim 1 and 2 is characterized in that described tumor is any one in leukemia, lymphoma, skin carcinoma, sarcoma, breast carcinoma, hepatocarcinoma, pulmonary carcinoma, glioma or the glioblastoma.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011103068750A CN102357100A (en) | 2011-10-12 | 2011-10-12 | Anti-tumor combination medicament |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011103068750A CN102357100A (en) | 2011-10-12 | 2011-10-12 | Anti-tumor combination medicament |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102357100A true CN102357100A (en) | 2012-02-22 |
Family
ID=45582518
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2011103068750A Pending CN102357100A (en) | 2011-10-12 | 2011-10-12 | Anti-tumor combination medicament |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102357100A (en) |
Cited By (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2017177947A1 (en) * | 2016-04-13 | 2017-10-19 | 诺马（北京）科技有限公司 | Method for treating pleuroperitoneal membrane cancers by locally injecting disulfiram preparation |
| CN108514639A (en) * | 2018-04-26 | 2018-09-11 | 武汉迈德森医药科技股份有限公司 | It is a kind of to be used for antitumor combination medicine and its preparation and application |
| CN109061185A (en) * | 2018-08-14 | 2018-12-21 | 兰州大学 | A method of copper carrier antitumor and anticancer agent is designed based on redox cycle strategy |
| CN109125346A (en) * | 2018-08-14 | 2019-01-04 | 兰州大学 | A kind of reproducible copper carrier antitumor and anticancer agent and its application |
| WO2019227530A1 (en) * | 2018-06-01 | 2019-12-05 | 北京好价盛行生物科技有限公司 | New anti-tumor use of disulfiram |
| US10695299B2 (en) | 2015-11-06 | 2020-06-30 | University Of Wolverhampton | Disulfiram formulation |
| CN111514122A (en) * | 2020-05-28 | 2020-08-11 | 青岛大学附属医院 | Application of disulfiram in preparation of drug for treating liposarcoma |
| CN112263591A (en) * | 2020-11-10 | 2021-01-26 | 中南大学 | Composition for treating cancer and application thereof |
| US11033579B2 (en) | 2015-09-24 | 2021-06-15 | Innolife Co., Ltd. | Use of trientine to deliver copper to ischemic tissue |
| CN114053418A (en) * | 2021-11-23 | 2022-02-18 | 中山大学 | Copper coordination nano-carrier, preparation method thereof and application of copper coordination nano-carrier as tumor chemotherapy drug |
| CN114269355A (en) * | 2019-05-21 | 2022-04-01 | 麦德赛尼克公司 | Use of metal ions to enhance the therapeutic effect of arsenic |
| CN114280308A (en) * | 2021-11-16 | 2022-04-05 | 中国科学院合肥物质科学研究院 | Application of Smad3 protein as novel anti-cisplatin drug-resistant target of ovarian cancer |
| CN114306414A (en) * | 2022-02-08 | 2022-04-12 | 东北林业大学 | Application of gamboge and disulfiram composition in preparation of medicine for treating colon cancer |
| CN114469995A (en) * | 2021-06-30 | 2022-05-13 | 四川大学 | Pharmaceutical composition for treating acute kidney injury and preparation method and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101217956A (en) * | 2005-05-05 | 2008-07-09 | 康宾纳特克斯公司 | Compositions and methods for treatment for neoplasms |
| CN102274347A (en) * | 2011-07-15 | 2011-12-14 | 陈迪 | Anticancer composition and application thereof |
-
2011
- 2011-10-12 CN CN2011103068750A patent/CN102357100A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101217956A (en) * | 2005-05-05 | 2008-07-09 | 康宾纳特克斯公司 | Compositions and methods for treatment for neoplasms |
| CN102274347A (en) * | 2011-07-15 | 2011-12-14 | 陈迪 | Anticancer composition and application thereof |
Non-Patent Citations (3)
| Title |
|---|
| DI CHEN，等: "Disulfiram, a Clinically used Anti-alcoholism drug and copper-binding agent,induces apoptotic cell death in breast cancer cultures and xenografts via inhibition of the proteasome activity", 《CANCER RESEARCH》 * |
| 史鹏程，等: "Disulfiram联合Cu对耐药白血病细胞的作用及与JNK通路的关系", 《广东医学》 * |
| 孙清，等: "Cu与DSF混合物抑制肿瘤细胞蛋白酶体活性的机理研究", 《中国药理学通报》 * |
Cited By (18)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US11033579B2 (en) | 2015-09-24 | 2021-06-15 | Innolife Co., Ltd. | Use of trientine to deliver copper to ischemic tissue |
| US10695299B2 (en) | 2015-11-06 | 2020-06-30 | University Of Wolverhampton | Disulfiram formulation |
| US10828271B2 (en) | 2016-04-13 | 2020-11-10 | University Of Wolverhampton | Method for treating pleuroperitoneal membrane cancers by locally injecting disulfiram preparation |
| WO2017177947A1 (en) * | 2016-04-13 | 2017-10-19 | 诺马（北京）科技有限公司 | Method for treating pleuroperitoneal membrane cancers by locally injecting disulfiram preparation |
| CN108514639A (en) * | 2018-04-26 | 2018-09-11 | 武汉迈德森医药科技股份有限公司 | It is a kind of to be used for antitumor combination medicine and its preparation and application |
| WO2019227530A1 (en) * | 2018-06-01 | 2019-12-05 | 北京好价盛行生物科技有限公司 | New anti-tumor use of disulfiram |
| CN109061185B (en) * | 2018-08-14 | 2022-07-15 | 兰州大学 | Method for designing copper carrier anti-cancer agent based on redox cycling strategy |
| CN109061185A (en) * | 2018-08-14 | 2018-12-21 | 兰州大学 | A method of copper carrier antitumor and anticancer agent is designed based on redox cycle strategy |
| CN109125346A (en) * | 2018-08-14 | 2019-01-04 | 兰州大学 | A kind of reproducible copper carrier antitumor and anticancer agent and its application |
| CN114269355B (en) * | 2019-05-21 | 2023-08-15 | 麦德赛尼克公司 | Use of metal ions to enhance the therapeutic effect of arsenic |
| CN114269355A (en) * | 2019-05-21 | 2022-04-01 | 麦德赛尼克公司 | Use of metal ions to enhance the therapeutic effect of arsenic |
| CN111514122A (en) * | 2020-05-28 | 2020-08-11 | 青岛大学附属医院 | Application of disulfiram in preparation of drug for treating liposarcoma |
| CN112263591A (en) * | 2020-11-10 | 2021-01-26 | 中南大学 | Composition for treating cancer and application thereof |
| CN114469995A (en) * | 2021-06-30 | 2022-05-13 | 四川大学 | Pharmaceutical composition for treating acute kidney injury and preparation method and application thereof |
| CN114280308A (en) * | 2021-11-16 | 2022-04-05 | 中国科学院合肥物质科学研究院 | Application of Smad3 protein as novel anti-cisplatin drug-resistant target of ovarian cancer |
| CN114053418A (en) * | 2021-11-23 | 2022-02-18 | 中山大学 | Copper coordination nano-carrier, preparation method thereof and application of copper coordination nano-carrier as tumor chemotherapy drug |
| CN114053418B (en) * | 2021-11-23 | 2023-11-14 | 中山大学 | Copper coordination nano-carrier, preparation method thereof and application of copper coordination nano-carrier as tumor chemotherapeutic medicine |
| CN114306414A (en) * | 2022-02-08 | 2022-04-12 | 东北林业大学 | Application of gamboge and disulfiram composition in preparation of medicine for treating colon cancer |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102357100A (en) | Anti-tumor combination medicament | |
| TWI574692B (en) | Use of sodium meta arsenite for the treatment of leukemia and lymphoma | |
| EP2786753B1 (en) | Combination therapy with an antitumor antibiotic | |
| EP3034076B1 (en) | Combined application of isothiocyanate compound and anti-cancer medicine | |
| US20110117210A1 (en) | Therapeutic treatment of human cancers using simple salts of zinc | |
| US20050107346A1 (en) | N-acetylcolchinol-o-phosphate combination therapies with vascular damaging activity | |
| CN102552908B (en) | Pharmaceutical composition containing artemisinin, artemisinin derivatives and Bcl-2 inhibitor and application thereof | |
| US9901602B2 (en) | Ejaculum of animals as medicinal material and uses thereof in medicaments for treatment of diseases such as tumors, depression, etc | |
| CN104434939B (en) | Antitumor medicament composition of panax notoginseng saponins R7 and oridonin with capabilities of reducing toxicity and enhancing efficacy and application of antitumor medicament composition | |
| KR101572529B1 (en) | Tetraarsenic oxide combination therapy for cancer treatment | |
| CN105380956B (en) | A kind of pharmaceutical composition of Dana Delany containing Chinese mugwort for treating leukaemia and application | |
| EP1389101B1 (en) | Toluene sulfonamide-containing anti-tumor composition and method of use thereof | |
| Vutakuri | Curcumin-breast cancer therapeutic agent to replace allopathic treatments with extensive side effects | |
| US10441564B2 (en) | Fructose analogs and their combinations as anti-cancer agents | |
| CN104800858A (en) | HSP90 inhibitory peptide conjugate and application thereof for treating tumours | |
| JPWO2015156409A1 (en) | Anticancer agent and side effect reducing agent | |
| CN102319260A (en) | The application of cisplatin combined itraconazole isomer in preparation treatment lung-cancer medicament | |
| US20200375943A1 (en) | Cytocidal method of cancer cells selectively in human patients clinically by depletion of l-ascorbic acid, primarily, with its supplementation alternately augmenting its cytocidal effect | |
| EP3949970A1 (en) | Combined use of a-nor-5? androstane compound drug and anticancer drug | |
| CN103054802A (en) | Procationic/ cationic liposome curcumin preparation for interventional treatment of hepatic carcinoma and preparation method of preparation | |
| CN104688748A (en) | Pharmaceutical composition containing ursolic acid and cyclophosphamide | |
| CN113164474A (en) | Water-soluble polymer derivative of Venetok | |
| CN104146999A (en) | Oridonin and docetaxel toxicity reducing and efficacy enhancing antitumor drug composition and application thereof | |
| CN102440987B (en) | Drug compound of apigenin, apigenin-like derivants, artemisinin and artemisinin-like derivants and application thereof | |
| CN110063989A (en) | A kind of pharmaceutical composition and preparation method thereof for treating the cancer of the esophagus |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C12 | Rejection of a patent application after its publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20120222 |