CN103285381A - A combination of ribonucleases and cantharidin - Google Patents
A combination of ribonucleases and cantharidin Download PDFInfo
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- CN103285381A CN103285381A CN2012100405386A CN201210040538A CN103285381A CN 103285381 A CN103285381 A CN 103285381A CN 2012100405386 A CN2012100405386 A CN 2012100405386A CN 201210040538 A CN201210040538 A CN 201210040538A CN 103285381 A CN103285381 A CN 103285381A
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Abstract
The invention relates to a combination of ribonucleases and cantharidin. Specifically, the invention discloses a pharmaceutical composition comprising: ribonucleases as an active component (a); cantharidin or derivatives or pharmaceutical acceptable salts of the cantharidin as an active component (b); and pharmaceutical acceptable carriers as active components (c). The pharmaceutical composition of the invention maycooperatively and effectively increase inhibition effects on the growth of tumor cells.
Description
Technical field
The invention belongs to biomedicine field.Particularly, the present invention relates to the coupling of ribonuclease and cantharidin.
Background technology
Ranpirnase (Ranp) is a kind of ribonuclease in the northern leopard frog (Rana pipiens) ovum or the body early embryo.It is made of 104 amino acid residues, about 12,000 dalton of molecular weight.It is a member of bovine pancreatic ribonuclease A (RNase A) superfamily, and it is 30% identical that its primary structure and RNase A have, and their tertiary structure is also rather similar.Yet, the most of members of the toxicity of the tumor cell of Ranpirnase in RNase A and the superfamily thereof, but little to normal cell damage.Ranpirnase can suppress the growth of kinds of tumors.Though Ranpirnase is pharmaceutical grade protein, immunogenicity is very low, can use a plurality of courses for the treatment of clinically.Its side reaction is little, and bone marrow is not had overt toxicity.Main side reaction is nephrotoxicity, but can reverse after the drug withdrawal.Ranpirnase Stability Analysis of Structures, phase transition temperature reach 90 degree.It has good stable in cell, the half-life is 3 hours in the body.The influence that the lethal effect of the tumor of Ranpirnase is not expressed by p53 state or P-gp, so Ranpirnase has the ability of anti-multi-drug resistant.
Cantharidin (cantharidin or exo, exo-2,3-dimethyl-7-oxabicyclo[2.2.1] heptane-2,3-dicarboxylic acid anhydride) is Meloidae (Mylabris) insecticide, a kind of natural toxin in Mylabris (Mylabris phalerata or the Mylabris cichorii) body belongs to the monoterpene compounds.Have now found that the cantharidin or derivatives thereof is the inhibitor of protein phosphatase 1 (PP1) and protein phosphatase 2a (PP2A).The inhibition of protein phosphatase can be blocked cell mitogen, and influences endocellular metabolism, and a series of processes such as signal conduction finally cause apoptosis.Different with many chemotherapeutics, the cantharidin medicine manifests the effect of leukocyte increasing quantity in the treatment tumor simultaneously.Cantharidin and its derivant clinically for the treatment of hepatocarcinoma, gastric cancer, pulmonary carcinoma, breast carcinoma and the esophageal carcinoma etc., show certain curative effect as disodium cantharidinate, norcantharidin, N-methylcantharidimide and N-hydroxycantharidin etc.
With the medicine of this two classes uniqueness, ribonuclease and cantharidin or cantharidin derivative or its pharmaceutically acceptable salt combine the treatment tumor, can produce what kind of effect, but are this area the unknowns, but press for understanding.
Summary of the invention
The present invention aims to provide ribonuclease and cantharidin or cantharidin derivative or its pharmaceutically acceptable salt coupling antineoplastic method.
First aspect present invention provides a kind of pharmaceutical composition, and described pharmaceutical composition contains:
Active component (a) ribonuclease;
Active component (b) cantharidin or cantharidin derivative or its pharmaceutically acceptable salt; And
(c) pharmaceutically acceptable carrier.
In another preference, described cantharidin or cantharidin derivative or its pharmaceutically acceptable salt are selected from down group: cantharidin, disodium cantharidinate, norcantharidin, Injectio natarii norcantharidatis, N-methylcantharidimide or N-hydroxycantharidin.
In another preference, described cantharidin or cantharidin derivative or its pharmaceutically acceptable salt are disodium cantharidinate.
In another preference, described ribonuclease is selected from down group: RNases, the Niu Jingguan RNase of leopard Rana ovum ribonuclease (Ranpirnase), the amphibious enzyme of leopard Rana ovum (Amphinase), bull frog or wood frog or the neurotoxin that people's eosinocyte is derived.
In another preference, described ribonuclease is leopard Rana ovum ribonuclease (Ranpirnase).
In another preference, the part by weight of described active component (a) and active component (b) is 30: 1 to 0.04: 1, preferably is 25: 1 to 0.04: 1.
In another preference, for the SPC-A-1 cell strain, the part by weight of described active component (a) and active component (b) is 1: 1 to 0.04: 1.
In another preference, for the A549 cell strain, the part by weight of described active component (a) and active component (b) is 10: 1 to 0.05: 1.
In another preference, the total content of described active component (a) and active component (b) is 0.0001～99.9wt% of compositions, and preferably 0.001～99wt% more preferably is 0.001～90wt%.
In another preference, the dosage form of described compositions is tablet, capsule, suppository, powder or injection (as intravenous fluid).
In another preference, the dosage form of described compositions is injection.
In another preference, described compositions is compound preparation.
In another preference, for the preparation of the medicine for the treatment of tumor or inhibition growth of tumour cell.
In another preference, described tumor is selected from down group: malignant mesothe, nonsmall-cell lung cancer, cancer of pancreas, nasopharyngeal carcinoma, thymoma, colon cancer, melanoma, renal carcinoma, leukemia, lymphoma, hepatocarcinoma, gastric cancer, esophageal carcinoma, breast carcinoma or skin carcinoma; Be preferably nonsmall-cell lung cancer.
Second aspect present invention provides a kind of combination that is made of active component (a) ribonuclease and active component (b) cantharidin or cantharidin derivative or its pharmaceutically acceptable salt or the purposes of mixture, for the preparation of the treatment tumor or suppress the medicine of growth of tumour cell.
Third aspect present invention provides a kind of medicine box, and described medicine box contains:
(i) first container, and be loaded on active component (a) ribonuclease in this first container or contain the medicine of active component (a);
(ii) second container, and be loaded on active component (b) cantharidin or cantharidin derivative or its pharmaceutically acceptable salt in this second container or contain the medicine of active component (b); And
(iii) description has been put down in writing in the described description to unite and has been given active component (a) and active component (b) thereby the explanation for the treatment of tumor.
Fourth aspect present invention provides a kind of medicine box, and described medicine box comprises:
The preparation of one qiagen rnase enzyme;
One contains the preparation of cantharidin or cantharidin derivative or its pharmaceutically acceptable salt; With
One description.
In another preference, the described preparation of cantharidin or cantharidin derivative or its pharmaceutically acceptable salt or the preparation of qiagen rnase enzyme of containing is selected from tablet, capsule, suppository, powder or injection, more preferably injection.
In another preference, described cantharidin or cantharidin derivative or its pharmaceutically acceptable salt are selected from down group: cantharidin, disodium cantharidinate, norcantharidin, Injectio natarii norcantharidatis, N-methylcantharidimide or N-hydroxycantharidin.
In another preference, described cantharidin or cantharidin derivative or its pharmaceutically acceptable salt are disodium cantharidinate.
In another preference, described ribonuclease is selected from down group: RNases, the Niu Jingguan RNase of leopard Rana ovum ribonuclease (Ranpirnase), the amphibious enzyme of leopard Rana ovum (Amphinase), bull frog or wood frog or the neurotoxin that people's eosinocyte is derived.
In another preference, described ribonuclease is leopard Rana ovum ribonuclease (Ranpirnase).
Fifth aspect present invention provides a kind of external non-therapeutic to suppress the method for growth of tumour cell, described method comprises step: at (a) ribonuclease with (b) in the presence of cantharidin or cantharidin derivative or its pharmaceutically acceptable salt, and culture of tumor cell.
In another preference, cell concentration is as every hole 4000-6000 cell preferred every hole 4500-5500 cell, more preferably 5000 cells in every hole; Incubation time is 48-96 hour.
In another preference, ribonuclease concentration is 0.0005-50 μ mol/L in the cultivating system, and the concentration of cantharidin or cantharidin derivative or its pharmaceutically acceptable salt is 0.05-20 μ mol/L.
In another preference, for the SPC-A-1 cell strain, ribonuclease concentration is 0.01-5 μ mol/L in the cultivating system, and the concentration of cantharidin or cantharidin derivative or its pharmaceutically acceptable salt is 0.05-20 μ mol/L.
In another preference, for the A549 cell strain, ribonuclease concentration is 0.10-50 μ mol/L in the cultivating system, and the concentration of cantharidin or cantharidin derivative or its pharmaceutically acceptable salt is 0.05-20 μ mol/L.
In another preference, described tumor cell is selected from down the cell of the tumor of group: malignant mesothe, nonsmall-cell lung cancer, cancer of pancreas, nasopharyngeal carcinoma, thymoma, colon cancer, melanoma, renal carcinoma, leukemia, lymphoma, hepatocarcinoma, gastric cancer, esophageal carcinoma, breast carcinoma or skin carcinoma etc.
In another preference, described tumor cell is nonsmall-cell lung cancer.
Sixth aspect present invention provides a kind of method for the treatment of tumor, and described method comprises step: use active component (a) ribonuclease and active component (b) cantharidin or cantharidin derivative or its pharmaceutically acceptable salt for the object (comprising mammal such as people) of needs; Perhaps use the pharmaceutical composition that contains described active component (a) and active component (b).
In should be understood that within the scope of the present invention, above-mentioned each technical characterictic of the present invention and can making up mutually between specifically described each technical characterictic in below (eg embodiment), thus constitute new or optimized technical scheme.As space is limited, this tired stating no longer one by one.
Description of drawings
Fig. 1 has shown that Ranp and SCA are for the associating Graft Versus Tumor of people's nonsmall-cell lung cancer SPC-A-1.
Fig. 2 has shown that Ranp and SCA are for the associating Graft Versus Tumor of people's nonsmall-cell lung cancer A-549.
The specific embodiment
The inventor is through extensive and deep research, and discovery ribonuclease and cantharidin or cantharidin derivative or its pharmaceutically acceptable salt are united use treatment tumor, can produce the obvious synergistic effect.Unite specifically and use leopard Rana ovum ribonuclease and disodium cantharidinate.On this basis, finished the present invention.
Ribonuclease
As used herein, " ribonuclease " refers to a kind of Cytotoxic ribonuclease that has, comprise leopard Rana ovum ribonuclease (Onconase, Onc or Ranpirnase, Ranp), the RNases of the amphibious enzyme of leopard Rana ovum (Amphinase), bull frog or wood frog, cattle spermaduct RNase, neurotoxin that people's eosinocyte is derived etc. and by the Pancreas Bovis seu Bubali of genetic engineering modified mistake or people's pancreas RNases etc.; Preferred Ranp.
Described leopard Rana ovum ribonuclease is a kind of for the useful enzyme of inhibition tumor cell, its tRNA that can in endochylema, optionally degrade, and Profilin is synthesized and then is suppressed cell proliferation and causes apoptosis.
In the present invention, used leopard Rana ovum ribonuclease can be naturally occurring, such as its can be separated or purification from animal.In addition, described leopard Rana ovum ribonuclease also can be artificial preparation, such as producing reorganization Rana ovum ribonuclease according to the genetic engineering recombinant technique of routine.Preferably, the present invention can adopt the leopard Rana ovum ribonuclease of reorganization.
Any suitable leopard Rana ovum ribonuclease all can be used for the present invention.Described leopard Rana ovum ribonuclease comprises leopard Rana ovum ribonuclease or its bioactive fragment of total length.
The aminoacid sequence of the leopard Rana ovum ribonuclease that passes through replacement, disappearance or the interpolation of one or more amino acid residues and form is also included among the present invention.Leopard Rana ovum ribonuclease or its bioactive fragment comprise the alternative sequence of a part of conserved amino acid, and described sequence of replacing through aminoacid does not influence its activity or kept the activity of its part.Suitably replacing aminoacid is the technique known of this area, and described technology can be implemented and guarantee not change the biological activity of gained molecule at an easy rate.These technology are recognized those skilled in the art, in general, can not change biological activity basically at the inessential area change single amino acids of a peptide species.See Mol ecular Biology of The Gene such as Watson, the 4th edition, 1987, The Benjamin/Cummings Pub.Co.P224.
The bioactive fragment of any leopard Rana ovum ribonuclease can be applied among the present invention.Here, the implication of the bioactive fragment of leopard Rana ovum ribonuclease refers to that as a peptide species it still can keep all or part of function of the leopard Rana ovum ribonuclease of total length.Generally, described bioactive fragment keeps the activity of 50% total length Rana ovum ribonuclease at least.Under preferred condition, described active fragment can keep 60%, 70%, 80%, 90%, 95%, 99% or 100% activity of total length leopard Rana ovum ribonuclease.
The present invention also can adopt leopard Rana ovum ribonuclease modified or improvement, such as, can adopt the leopard Rana ovum ribonuclease of being modified or improveing in order to prolong its half-life, improve its stability or to strengthen its killing tumor cell ability.Described through modifying or though leopard Rana ovum ribonuclease or the gene of improvement can be to have certain difference with naturally occurring leopard Rana ovum ribonuclease or gene, also can killing tumor cell, and can not bring other harmful effect or toxicity.That is to say that the version of the biological function of any biological activity that does not influence leopard Rana ovum ribonuclease or perhaps gene all can be used among the present invention.
As an example of the present invention, described leopard Rana ovum ribonuclease is recombinant expressed, the encoding gene total length 315bp of described leopard Rana ovum ribonuclease, the polypeptide of 104 AA of coding.
In general described leopard Rana ovum ribonuclease can have following steps by conventional gene engineering method production:
(1) with the recombinant expression carrier conversion that contains leopard Rana ovum ribonucleic acid enzyme coding gene or transduction proper host cell;
(2) host cell of in proper culture medium, cultivating;
(3) separation, protein purification from culture medium or cell.
Cantharidin or cantharidin derivative or its pharmaceutically acceptable salt
As used herein, " cantharidin or cantharidin derivative or its pharmaceutically acceptable salt " comprises cantharidin, disodium cantharidinate, norcantharidin, Injectio natarii norcantharidatis, N-methylcantharidimide and N-hydroxycantharidin etc.; Preferred disodium cantharidinate (SCA).
Below be several known cantharidin in this area or the chemical formula of cantharidin derivative or its pharmaceutically acceptable salt:
Cantharidin norcantharidin disodium cantharidinate Injectio natarii norcantharidatis
The disodium cantharidinate (SCA) that the present invention adopts is made through chemistry its anhydride that ftractures by cantharidin.
" cantharidin derivative " as herein described in literary composition, mention those, comprise that also those skilled in the art can be by simple chemical method, for example replace, method such as reduction modifies for the side-chain radical on the skeleton, and still keeps those derivants of cantharidin activity.The synthetic method of these derivants can be with reference to textbook of the prior art.Simultaneously, cantharidin derivative also comprises salt, hydrate, optical isomer of its various soda acid forms etc.
As used herein, term " pharmaceutically acceptable salt " refers to the formed salt that is suitable as medicine of The compounds of this invention and acid or alkali.Pharmaceutically acceptable salt comprises inorganic salt and organic salt.The preferred salt of one class is the salt that The compounds of this invention and alkali form.The alkali that is fit to formation salt includes, but are not limited to: sodium hydroxide etc.The preferred salt of one class is The compounds of this invention and the sour salt that forms.The acid that is fit to formation salt includes, but are not limited to: mineral acids such as hydrochloric acid, organic acid such as formic acid, acetic acid; And, acidic amino acid such as glutamic acid.
Pharmaceutical composition and medicine box
The invention provides and contain active component (a) ribonuclease; Active component (b) cantharidin or cantharidin derivative or its pharmaceutically acceptable salt; And (c) pharmaceutical composition of pharmaceutically acceptable carrier.
Dosage form and the preparation method of pharmaceutical composition of the present invention are not particularly limited, and the conventional general method for making in available this area is made various dosage forms such as injection (as intravenous fluid) or masking liquid, suppository, powder.A kind of preferred dosage form is injection (as intravenous fluid).
The present invention's used " pharmaceutically acceptable carrier " refers to: one or more compatibility solids or liquid filler or gelatinous mass, they are suitable for the people uses, and enough purity and enough low toxicity must be arranged." compatibility " referred to herein as each component energy and chemical compound of the present invention and blending mutually between them in the compositions, and the drug effect of not obvious reduction chemical compound.Pharmaceutically acceptable carrier part example has cellulose and derivant (as sodium carboxymethyl cellulose, ethyl cellulose sodium, cellulose ethanoate etc.) thereof, gelatin, Talcum, kollag (as stearic acid, magnesium stearate), calcium sulfate, vegetable oil (as Oleum Glycines, Oleum sesami, Oleum Arachidis hypogaeae semen, Fructus Canarii albi wet goods), polyhydric alcohol (as propylene glycol, glycerol, mannitol, sorbitol etc.), emulsifying agent (as tween
), wetting agent (as sodium lauryl sulphate), coloring agent, flavoring agent, stabilizing agent, antioxidant, antiseptic, apirogen water etc.
Preferably, pharmaceutical composition of the present invention is compound preparation, and it comprises active component (a) ribonuclease in safety and the treatment effective dose scope; Active component (b) cantharidin or cantharidin derivative or its pharmaceutically acceptable salt; And (c) pharmaceutically acceptable carrier.
As used herein, term " safety and treatment effective dose " refers to: the amount of active component is enough to obviously improve the state of an illness, and is unlikely to produce serious adverse.Usually, compound preparation contains the active component/agent of the present invention of 1-2000 microgram, more preferably, contains the active component/agent of the present invention of 10-1000 microgram.Preferably, described " potion " is an ampoule (injection bottle).
Active component of the present invention (a) ribonuclease is preferably by the injection system administration; Active component (b) can be by injection or other mode administrations.
The preferred powder of compound preparation of the present invention (as freeze dried powder) or injection more preferably are injection.
For active component (a) ribonuclease, be preferably liquid dosage form or be reconfigurable into liquid dosage form.Preparation liquid dosage form of the present invention comprises pharmaceutically acceptable emulsion, solution or suspension.Except the active ingredient beyond the region of objective existence, liquid dosage form can comprise the conventional inert diluent that adopts in this area, as water or other solvent, solubilizing agent and emulsifying agent, example is known, ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3 butylene glycol, dimethyl formamide etc.Except these inert diluents, compositions also can comprise auxiliary agent, as wetting agent, emulsifying agent and suspending agent (for example, the mixture of ethoxylation isooctadecane alcohol, polyoxyethylene sorbitol and Isosorbide Dinitrate, microcrystalline Cellulose, aluminium methoxide and agar or these materials) etc.
The preparation that is used for parenteral injection can comprise physiologically acceptable aseptic moisture or anhydrous solution, dispersion liquid, suspension or emulsion and be used for being dissolved into again aseptic Injectable solution or the sterilized powder of dispersion liquid.Suitable moisture and nonaqueous carrier, diluent, solvent or excipient comprise water, ethanol, polyhydric alcohol and suitable mixture thereof.
Yet, for the compositions that only contains active component (b) cantharidin or cantharidin derivative, can be for solid dosage forms or liquid dosage form (or reconstituting liquid dosage form), comprising capsule, tablet, pill, powder (or powder), granule and injection; Preferably tablet, powder or injection more preferably are injection.
Solid dosage forms such as tablet, sugar pill, capsule, pill and granule can adopt coating and the preparation of shell material, as casing and other material well known in the art.They can comprise opacifying agent, and, discharge in the mode that the release of active component can postpone in this preparation certain part in digestive tract.The example of adoptable embedding component is polymeric material and Wax.In case of necessity, active component also can with above-mentioned excipient in one or more form microencapsulation form.
Transdermal administration also is one of preferred formulation, and the dosage form that is used for the The compounds of this invention of transdermal administration comprises micropin agent and patch etc.Active component under aseptic condition with physiologically acceptable carrier, or the propellant that may need in case of necessity is mixed together.
The present invention also provides a kind of medicine box that can be used for treating tumor, and this medicine box contains:
(i) first container, and be loaded on active component (a) ribonuclease in this first container or contain the medicine of active component (a);
(ii) second container, and be loaded on active component (b) cantharidin or cantharidin derivative or its pharmaceutically acceptable salt in this second container or contain the medicine of active component (b); And
(iii) description has been put down in writing in the described description to unite and has been given active component (a) and active component (b) thereby the explanation for the treatment of tumor.
In another preference, described medicine is the single preparations of ephedrine that contains the single preparations of ephedrine of active component (a) or contain active component (b).
In another preference, active component (a) ribonuclease or the medicine that contains active component (a) are preferably injection; Active component (b) cantharidin or cantharidin derivative or its pharmaceutically acceptable salt or the medicine that contains active component (b) are tablet or injection.
The present invention also provides a kind of medicine box, and described medicine box comprises:
One contains the preparation of cantharidin or cantharidin derivative or its pharmaceutically acceptable salt;
The preparation of one qiagen rnase enzyme; With
One description.
Described preparation preferred tablet or the injection that contains cantharidin or cantharidin derivative or its pharmaceutically acceptable salt preferably is injection.
The preparation optimizing injection of described qiagen rnase enzyme.
The described preparation that contains cantharidin or cantharidin derivative or its pharmaceutically acceptable salt can be the unit dosage form that contains cantharidin or cantharidin derivative or its pharmaceutically acceptable salt, and the preparation of described ribonuclease can be the unit dosage form of qiagen rnase enzyme.
Be equipped with at least two in the medicine box and contain the unit dosage form of cantharidin or cantharidin derivative or its pharmaceutically acceptable salt and the unit dosage form of at least two qiagen rnase enzymes; It preferably respectively is 4-10.
As used herein, term " unit dosage form " refers to for taking convenience, becomes single to take required dosage form preparation of compositions, includes but not limited to various solid formulation (as tablet), liquid agent (as injection), capsule, slow releasing agent.
Describedly containing the unit dosage form of cantharidin or cantharidin derivative or its pharmaceutically acceptable salt or the unit dosage form of described qiagen rnase enzyme can be tablet, powder or injection, preferably is injection.
Following description is arranged in the description provided by the invention: the using method of described medicine box is to use the unit dosage form of qiagen rnase enzyme and contains the unit dosage form of cantharidin or cantharidin derivative or its pharmaceutically acceptable salt, every one day, repeat such occupation mode once.
Medicine box provided by the invention prepares by conventional method, for example: will contain the preparation of cantharidin or cantharidin derivative or its pharmaceutically acceptable salt and the preparation of qiagen rnase enzyme, and description is placed the formation medicine box together.The described preparation that contains cantharidin or cantharidin derivative or its pharmaceutically acceptable salt preferably contains the unit dosage form of cantharidin or cantharidin derivative or its pharmaceutically acceptable salt, the unit dosage form of the preferred qiagen rnase enzyme of the preparation of described ribonuclease.
Medicine box provided by the invention or compound medicament composition are used for the treatment of tumor, and described tumor comprises malignant mesothe, nonsmall-cell lung cancer, cancer of pancreas, nasopharyngeal carcinoma, thymoma, colon cancer, melanoma, renal carcinoma and leukemia and lymphoma, hepatocarcinoma, gastric cancer, esophageal carcinoma, breast carcinoma, skin carcinoma etc.; Preferred nonsmall-cell lung cancer.
External non-therapeutic suppresses the method for growth of tumour cell
The invention provides the method that a kind of external non-therapeutic suppresses growth of tumour cell, described method comprises step: at (a) ribonuclease with (b) in the presence of cantharidin or cantharidin derivative or its pharmaceutically acceptable salt, and culture of tumor cell.
Described tumor cell comprises malignant mesothe, nonsmall-cell lung cancer, cancer of pancreas, nasopharyngeal carcinoma, thymoma, colon cancer, melanoma, renal carcinoma and leukemia and lymphoma, hepatocarcinoma, gastric cancer, esophageal carcinoma, breast carcinoma, skin carcinoma etc.; Preferred nonsmall-cell lung cancer.
In another preference, cell concentration is as every hole 4000-6000 cell preferred every hole 4500-5500 cell, more preferably 5000 cells in every hole; Incubation time is 48-96 hour, preferred 60-80 hour.
Major advantage of the present invention
1.Ranp have synergism with the SCA coupling, can strengthen the anticancer growth.
2. the side reaction of two medicines is not quite similar, and during coupling, the amount that the dosage of each medicine uses more separately is low, therefore can alleviate side reaction effectively.
3.Ranp with SCA all be hypotoxicity, the medicine of reduced immunogenicity, their combination should keep above-mentioned characteristic, and might life-time service.
4.Ranp do not damage bone marrow with SCA, do not cause alopecia yet, different with the mechanism of action of other conventional cancer therapy drug, may be on one's body the patient of conventional chemotherapy failure.
Below in conjunction with specific embodiment, further set forth the present invention.Should be understood that these embodiment only to be used for explanation the present invention and be not used in and limit the scope of the invention.The experimental technique of unreceipted actual conditions in the following example, usually according to normal condition, people such as Sambrook for example, molecular cloning: laboratory manual (New York:Cold Spring Harbor Laboratory Press, 1989) condition described in, or the condition of advising according to manufacturer.Unless otherwise indicated, otherwise all percentage ratio and umber by weight.
Unit in the percent weight in volume among the present invention is well-known to those skilled in the art, for example refers to the weight of solute in 100 milliliters solution.
Unless otherwise defined, the same meaning that employed all specialties and scientific words and one skilled in the art are familiar with in the literary composition.In addition, any method similar or impartial to described content and material all can be applicable in the inventive method.The usefulness that preferable implementation method described in the literary composition and material only present a demonstration.
Embodiment 1 unites the cytotoxic effect that uses Ranp and the people's nonsmall-cell lung cancer of SCA SPC-A-1 cell
The SPC-A-1 cell culture is in containing 10% hyclone, and penicillin (100units/ml) is in RPMI1640 (GIBICO) culture medium of streptomycin (100 μ g/ml).96 orifice plates are cultivated, and every hole adds 90 μ l culture fluid (5 * 10
4Cells/ml), CO
2(5%) cultivate 12h in the incubator after, add the Ranp-SCA blend sample 10 μ l (as shown in table 1) of the various proportionings of setting Concentraton gradient, disposable individually dosed Ranp, SCA and the PBS of adding respectively in the matched group hole in the administering drug combinations hole respectively.After cultivating 72h, mtt assay detects cell survival rate, utilizes medicine equivalent line rule to judge its administering drug combinations effect.Measurement result is shown in Fig. 1 and table 1.
Table 1
Fig. 1 has shown that Ranp and SCA are for the associating Graft Versus Tumor of people's nonsmall-cell lung cancer SPC-A-1.By Ranp and SCA IC separately in every kind of combination in the last table
50Value, value with Ranp in all proportions is X-axis, the value of SCA is that Y-axis obtains two-dimensional coordinate figure, ratio is respectively the coordinate points line (addition line) of [5/0] and [0/5], because 4 left sides that are positioned at line of residue, therefore reach a conclusion: Ranp and SCA have synergism for the administering drug combinations of people's nonsmall-cell lung cancer SPC-A-1, and the anticancer effect of two medicine couplings uses obvious enhancing respectively than single medicine.
Embodiment 2 unites the cytotoxic effect that uses Ranp and the people's nonsmall-cell lung cancer of SCA A-549 cell
The A-549 cell culture is in containing 10% hyclone, and penicillin (100units/ml) is in RPMI 1640 (GIBICO) culture fluid of streptomycin (100 μ g/ml).96 orifice plates are cultivated, and every hole adds 90 μ l culture fluid (5 * 10
4Cells/ml), CO
2(5%) cultivate 12h in the incubator after, add the Ranp-SCA blend sample 10 μ l (as shown in table 2) of the various proportionings of setting Concentraton gradient, disposable individually dosed Ranp, SCA and the PBS of adding respectively in the matched group hole in the administering drug combinations hole respectively.After cultivating 72h, mtt assay detects cell survival rate, utilizes medicine equivalent line rule to judge its administering drug combinations effect.Measurement result is shown in Fig. 2 and table 2.
Table 2
Fig. 2 has shown that Ranp and SCA are for the associating Graft Versus Tumor of people's nonsmall-cell lung cancer A-549.By Ranp and SCA IC separately in every kind of combination in the last table
50Value, value with Ranp in all proportions is X-axis, the value of SCA is that Y-axis obtains two-dimensional coordinate figure, ratio is respectively the coordinate points line (addition line) of [5/0] and [0/5], because 4 left sides that are positioned at line of residue, therefore reach a conclusion: Ranp and SCA have synergism for the administering drug combinations of people's nonsmall-cell lung cancer A549, and the anticancer effect of two medicine couplings uses obvious enhancing respectively than single medicine.
Experimental results show that:
Unite and use ribonuclease (as Ranpirnase) and cantharidin or cantharidin derivative or its pharmaceutically acceptable salt (as disodium cantharidinate etc.) can obviously strengthen the effect of inhibition growth of tumour cell.The coupling of two kinds of medicines has the obvious synergistic effect.During two kinds of drug combinations, the amount that the dosage of each medicine uses more separately is low, therefore can alleviate side reaction effectively.
Pharmaceutical composition
Active component (a) Ranp 500 μ g
Active component (b) SCA 25 μ g
Be injection with above-mentioned two kinds of active component mixed configuration, place an ampoule (specification 2ml), stand-by.
Embodiment 4
Pharmaceutical composition
Active component (a) Ranp 500 μ g
Active component (b) SCA 25 μ g
Ranp is configured to injection with active component (a), places an ampoule (specification 1ml), and is stand-by; SCA is configured to injection with active component (b), places an ampoule (specification 1ml), and is stand-by.
All quote in this application as a reference at all documents that the present invention mentions, just quoted as a reference separately as each piece document.Should be understood that in addition those skilled in the art can make various changes or modifications the present invention after having read above-mentioned teachings of the present invention, these equivalent form of values fall within the application's appended claims institute restricted portion equally.
Claims (10)
1. a pharmaceutical composition is characterized in that, described pharmaceutical composition contains:
Active component (a) ribonuclease;
Active component (b) cantharidin or cantharidin derivative or its pharmaceutically acceptable salt; And
(c) pharmaceutically acceptable carrier.
2. pharmaceutical composition as claimed in claim 1, it is characterized in that described cantharidin or cantharidin derivative or its pharmaceutically acceptable salt are selected from down group: cantharidin, disodium cantharidinate, norcantharidin, Injectio natarii norcantharidatis, N-methylcantharidimide or N-hydroxycantharidin.
3. pharmaceutical composition as claimed in claim 1, it is characterized in that described ribonuclease is selected from down group: RNases, the Niu Jingguan RNase of leopard Rana ovum ribonuclease (Ranpirnase), the amphibious enzyme of leopard Rana ovum (Amphinase), bull frog or wood frog or the neurotoxin that people's eosinocyte is derived.
4. pharmaceutical composition as claimed in claim 1 is characterized in that, the part by weight of described active component (a) and active component (b) is 30: 1 to 0.04: 1, preferably is 25: 1 to 0.04: 1.
5. pharmaceutical composition as claimed in claim 1 is characterized in that, the total content of described active component (a) and active component (b) is 0.0001～99.9wt% of compositions, preferably is 0.001～99wt%.
6. pharmaceutical composition as claimed in claim 1 is characterized in that, the dosage form of described compositions is tablet, capsule, suppository, powder or injection (as intravenous fluid).
7. the purposes of pharmaceutical composition as claimed in claim 1 is characterized in that, for the preparation of the medicine for the treatment of tumor or inhibition growth of tumour cell.
8. a combination that is constituted by active component (a) ribonuclease and active component (b) cantharidin or cantharidin derivative or its pharmaceutically acceptable salt or the purposes of mixture, it is characterized in that, for the preparation of the medicine for the treatment of tumor or inhibition growth of tumour cell.
9. a medicine box is characterized in that, described medicine box contains:
(i) first container, and be loaded on active component (a) ribonuclease in this first container or contain the medicine of active component (a);
(ii) second container, and be loaded on active component (b) cantharidin or cantharidin derivative or its pharmaceutically acceptable salt in this second container or contain the medicine of active component (b); And
(iii) description has been put down in writing in the described description to unite and has been given active component (a) and active component (b) thereby the explanation for the treatment of tumor;
Perhaps, described medicine box comprises:
The preparation of one qiagen rnase enzyme;
One contains the preparation of cantharidin or cantharidin derivative or its pharmaceutically acceptable salt; With
One description.
10. an external non-therapeutic suppresses the method for growth of tumour cell, it is characterized in that described method comprises step: at (a) ribonuclease with (b) in the presence of cantharidin or cantharidin derivative or its pharmaceutically acceptable salt, and culture of tumor cell.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210040538.6A CN103285381B (en) | 2012-02-22 | 2012-02-22 | A combination of ribonucleases and cantharidin |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201210040538.6A CN103285381B (en) | 2012-02-22 | 2012-02-22 | A combination of ribonucleases and cantharidin |
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| CN103285381A true CN103285381A (en) | 2013-09-11 |
| CN103285381B CN103285381B (en) | 2015-06-24 |
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| CN201210040538.6A Active CN103285381B (en) | 2012-02-22 | 2012-02-22 | A combination of ribonucleases and cantharidin |
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Citations (4)
| 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 |
| CN101775400A (en) * | 2009-01-14 | 2010-07-14 | 北京联合大学生物化学工程学院 | Rana chensinensis functional gene Rd-RNase2 sequence, construction method and amino acid sequence and application thereof |
| CN102188698A (en) * | 2010-03-08 | 2011-09-21 | 上海南方模式生物科技发展有限公司 | Combination of ribonuclease and artemisinin |
| CN102526714A (en) * | 2011-08-24 | 2012-07-04 | 贵州神奇集团控股有限公司 | Medicine composition for curing tumour and preparation method thereof |
-
2012
- 2012-02-22 CN CN201210040538.6A patent/CN103285381B/en active Active
Patent Citations (4)
| 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 |
| CN101775400A (en) * | 2009-01-14 | 2010-07-14 | 北京联合大学生物化学工程学院 | Rana chensinensis functional gene Rd-RNase2 sequence, construction method and amino acid sequence and application thereof |
| CN102188698A (en) * | 2010-03-08 | 2011-09-21 | 上海南方模式生物科技发展有限公司 | Combination of ribonuclease and artemisinin |
| CN102526714A (en) * | 2011-08-24 | 2012-07-04 | 贵州神奇集团控股有限公司 | Medicine composition for curing tumour and preparation method thereof |
Non-Patent Citations (2)
| Title |
|---|
| ANTONELLO MERLINO ET AL.: "Structural features for the mechanism of antitumor action of a dimeric human pancreatic ribonuclease variant", 《PROTEIN SCIENCE》, vol. 18, 2 December 2008 (2008-12-02), pages 50 - 57 * |
| 翟昌宝等: "豹蛙核酸酶与斑蝥酸钠对肺腺癌细胞增殖的协同抑制作用", 《中国细胞生物学学报》, vol. 35, no. 3, 31 December 2013 (2013-12-31), pages 328 - 333 * |
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| CN103285381B (en) | 2015-06-24 |
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Address after: 550004 No. 1, Beijing Road, Yunyan District, Guizhou, Guiyang Co-patentee after: Shanghai biomodel Polytron Technologies Inc Patentee after: Guizhou Maqika Group Address before: 550004 No. 1, Beijing Road, Yunyan District, Guizhou, Guiyang Co-patentee before: Shanghai Biomodel Organism Science & Technology Co.,Ltd. Patentee before: Guizhou Maqika Group |