CN102406649A - Application of five normal basic groups of human body in preparation of medicines for treating tumors - Google Patents
Application of five normal basic groups of human body in preparation of medicines for treating tumors Download PDFInfo
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- CN102406649A CN102406649A CN2011103613136A CN201110361313A CN102406649A CN 102406649 A CN102406649 A CN 102406649A CN 2011103613136 A CN2011103613136 A CN 2011103613136A CN 201110361313 A CN201110361313 A CN 201110361313A CN 102406649 A CN102406649 A CN 102406649A
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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
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
- A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
- A61K31/513—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim having oxo groups directly attached to the heterocyclic ring, e.g. cytosine
-
- 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
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
- A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
- A61K31/519—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with heterocyclic rings
- A61K31/52—Purines, e.g. adenine
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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
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
- A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
- A61K31/519—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with heterocyclic rings
- A61K31/52—Purines, e.g. adenine
- A61K31/522—Purines, e.g. adenine having oxo groups directly attached to the heterocyclic ring, e.g. hypoxanthine, guanine, acyclovir
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/16—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
- A61K9/1605—Excipients; Inactive ingredients
- A61K9/1629—Organic macromolecular compounds
- A61K9/1641—Organic macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyethylene glycol, poloxamers
- A61K9/1647—Polyesters, e.g. poly(lactide-co-glycolide)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
- A61K9/4841—Filling excipients; Inactive ingredients
- A61K9/4866—Organic macromolecular compounds
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
Abstract
The invention discloses application of five normal basic groups of a human body in preparation of medicines for treating tumors. The five basic groups are applied to the field of tumor treatment and can be prepared into orally taken preparation, injection, controlled release preparation or targeted preparation. Comparison with a standard product in anti-tumor effect shows that the five normal basic groups participating in human metabolites have functions of inducing tumor apoptosis and resisting tumor angiogenesis to some extent when reaching a certain concentration in vivo and in vitro. The invention has the beneficial effects that the five basic groups have broad-spectrum low-toxicity anti-tumor effect as compared with other anti-tumor medicines, can be used for chemotherapy of gastric cancer, lung cancer, liver cancer, colon cancer, breast cancer, reproductive system malignant tumor and the like and have obvious anti-tumor effect, no bone marrow suppression or main organ injury; the dosage of the five basic groups accords with convention; in addition, the anti-tumor effect of the five basic groups is enhanced as compared with that of normal nucleoside.
Description
Technical field
The present invention relates to a kind of antitumor drug, specifically five of body metabolism kinds of normal base application in preparation medicine for treating tumor thing belong to the tumour medicine field.
Background technology
The normal base of people comprises adenine (Adenine) CAS:73-24-5, molecular formula: C5H5N5, molecular weight: 135.13; Guanine (Guanine) CAS:73-40-5, molecular formula: C5H5N5O, molecular weight: 151.13; Cytosine (Cytosine) CAS:71-30-7, molecular formula: (C4H5N3O), molecular weight: 111.11; Thymus pyrimidine (Thymin) CAS:65-71-4, molecular formula: C5H6N2O2, molecular weight: 126.11; Uracil (Uracil) CAS:66-22-8, molecular formula: C4H4N2O2, molecular weight: 112.09.
Base many forms with nucleoside monophosphate in DNA and RNA exist, but the formation of its pair relationhip depends primarily on base.The normal base of people is digested and assimilated in the process and can be produced at nucleic acid material, also is that nucleic acid material is remedied synthetic important source material.But normal nucleotide metabolism can occur directly forming metabolic waste without base to excrete, as: adenosine → inosine → xanthosine → uric acid excretes.
More than five kinds of bases all can obtain at the external use distinct methods, be important medicine and health product intermediate.Wherein adenine is claimed adenine phosphate again, participates in RNA and DNA in vivo and synthesizes.When leukocyte lacked, it can promote leucocyte hyperplasia, and in general 2～4 weeks of medication, the leukocyte number can increase, and is used for the medicine of leukocyte increasing when being the Patients Receiving Chemotherapy leucocytes reduction.The adenine of one of five kinds of bases of human body be because can participate in the synthetic of nucleic acid such as DNA and RNA, can be in materials such as synthetic ATP its important function, present adenine is claimed adenine phosphate again, can be used for the treatment of the leukopenia due to the tumour patient chemotherapy etc.The external preservation that can be used for blood.
At present, universally recognized treatment tumor theory is in academia: through on normal base, adding different elements or substituent group, form normal base analogue and block the synthetic of cell DNA or RNA, thereby suppress the diffusion of tumor.For example; Name is called " Synergistic anti-cancer compounds ", the patent No. is the patent application of " CN200480036362.0 "; Use antimetabolic base analogue is disclosed as the antineoplastic composition; Anti-tumor metabolism base analogue is through suppressing the nucleus synzyme, and the growth of inhibition or prophylaxis of cancer and interference cell nucleic acid are synthetic.
Summary of the invention
The objective of the invention is to, the application of five kinds of normal bases of human body in preparation medicine for treating tumor thing is provided, these five kinds of bases are respectively adenine, guanine, cytosine, thymus pyrimidine and uracil; Above-mentioned five kinds of bases are processed oral formulations, injection, slow releasing agent or targeting preparation; Anti-curing oncoma effect is obvious; And multiple base combination is used as the ingredient of treatment tumor; Application can reach obvious raising antitumous effect separately, reduces toxic and side effects and delays the purpose that drug resistance takes place.
Technical scheme of the present invention is:
The application of five kinds of normal bases of human body in preparation medicine for treating tumor thing, described base comprises: adenine, guanine, cytosine, thymus pyrimidine and uracil.
Preferably, described base is one or more in adenine, guanine, cytosine, thymus pyrimidine and the uracil;
Preferably, described base is the combination of one or both and adenine in guanine, cytosine, thymus pyrimidine or the uracil.
Described medicine is oral tablet, injection or slow releasing agent.
The method for preparing of described oral tablet is: described base is added different auxiliary material such as starch; The part by weight of base and adjuvant is 1:1; Process oral tablet; Described base total content is the 10-30mg/ sheet, and the weight size of tablet for example can be 1g, 2g or 5g etc., can not exert an influence to its treatment and use;
The method for preparing of described injection is: described base is dissolved in the phosphate buffer solution that concentration is 5-25mg/mL, processes the injection that the base total concentration is 5-25mg/mL, every injection volume is 2-10mL;
The method for preparing of described slow releasing agent is: medicament liposome, microcapsule or micelle medicine-releasing system are processed the 5-25mg/mL slow releasing agent with described base, then slow releasing agent are made injection.
Further, the molal quantity of above-mentioned each base combination is identical.
Described tumor comprises multiple common cancers such as gastric cancer, pulmonary carcinoma, hepatocarcinoma, colon cancer, breast carcinoma, leukemia, genital system tumor.
Through discovering: guanine, cytosine, thymus pyrimidine and uracil antitumor action are lower than adenine, because of the side effect of adenine very I and the combination of other base as antineoplastic agent.Different adenine composite reagents reduces than the single medicine concentration in the single drug blood, thereby toxic and side effects descends.Multiple medicine composite reagent can delay the generation of drug resistance of tumor simultaneously.Through composite reagent, the dosage in the time of can reducing each composition single drug, and the absolute concentration of every kind of concrete nucleoside obviously descends, medicine just descends to the toxic and side effects of body; Simultaneously, two kinds or multiple chemotherapeutics composite reagent can reduce chemical sproof generation greatly.With adenosine and other people's normal alkali is basis set close after, be used in the medicine of the above-mentioned tumor of preparation treatment, can add adjuvant and process oral formulations or add its dissolubility of increase such as buffer salt and process injection, inject use; Injection can be conventional injection, also various types of slow releasing injection.
Different tumors have different nucleic acid metabolisms and compositing characteristic, every kind of base of different tumor cells is had different inhibition strengths, but the antitumor action of adenine are the strongest, and when experiment in vitro reached 0.3mg/mL when concentration, tumor control rate was more than 90% in 72 hours.30% of single adenine treatment tumor tumor body size is alleviated fully, and effective percentage is more than 93.0%.
Using dosage is low, is 0.001～0.05mmol/Kg/ days, intramuscular injection or intravenous drip; Oral administration can increase dose 0.5-1.5 doubly.
The invention has the advantages that:
1, adenine and other people's normal alkali are basis set share in the medicine of multiple common cancers such as preparation treatment gastric cancer, pulmonary carcinoma, hepatocarcinoma, colon cancer, breast carcinoma, genital system tumor, and antitumous effect is obvious, has the characteristics of efficient, wide spectrum, low toxicity;
2, adenine and the basis set antitumor action that closes of other people's normal alkali are apparently higher than all kinds of nucleoside and combination thereof;
3, adenine reduces than the single medicine concentration in the single drug blood with the basis set medicine that share of other people's normal alkali, thereby toxic and side effects reduces;
4, adenine and other people's normal alkali are basis set share the generation that medicine can delay drug resistance of tumor.
The specific embodiment
Below the preferred embodiments of the present invention are described, should be appreciated that preferred embodiment described herein only is used for explanation and explains the present invention, and be not used in qualification the present invention.
Embodiment 1
The adenine of variable concentrations and 5-FU be external to press down the tumor control experiment.
One, test method
With human cervical carcinoma Hela tumor cell (Chinese Academy of Sciences's Shanghai cell provides), with containing RPMI 1640 culture medium culturings of 10% NBCS, the cell of the trophophase of taking the logarithm, with behind 0.25% trypsinization with 3 * 10
3Individual/hole is inoculated in 96 orifice plates, places 37 ℃, 5%CO
2Cultivated 24 hours in the incubator; Establish 8 multiple holes for every group; Test group establishes respectively that multiple hole drug level is 0.005,0.01,0.02,0.04,0.08,0.16,0.32,0.64mg/mL totally 8 total concentrations, the every hole 200 μ L of culture fluid, and the 5-FU of matched group is identical with test group concentration; With serum-free medium dilution, the aseptic filtering with microporous membrane of 0.22 μ m.The Measuring Time of drafting 48 hours, inverted microscope observation of cell form was also taken pictures, and every then hole adds the MTT solution 20 μ L of 5mg/mL, places 37 ℃, 5%CO
2Continue in the incubator to cultivate 4 hours, stop cultivating, inhale and abandon the supernatant of cultivating in the hole; Every hole adds 150 μ L DMSO, shakes 10 minutes, crystal is fully dissolved after; Measure 570mn place absorbance with automatic ELIASA (Thermo, model MULTISKAN MK3).Each plate is established 8 multiple holes for every group, each experiment repetition 3 times, and data are the average of three experiments in the table 1.
Growth of tumour cell suppression ratio computing formula is following:
The OD value of the actual OD value/negative control hole in tumor cell survival rate (%)=dosing hole;
Growth of tumour cell suppression ratio (%)=100%-cell survival rate.
Table 1: adenine is to 48 hours suppression ratio (%)
of Hela cell
Wherein: Adenine is an adenine, and 5-FU is a 5-fluorouracil.
Two, result and analysis
Through calculate adenine and 5-FU in 0.005 to 0.64 concentration range 48 hours to Hela cell inhibiting rate no difference of science of statistics (
P>0.5); Adenine IC50 is about 70ug/mL.
Embodiment 2
The external tumor that presses down of five kinds of normal bases of people is tested.
One, test method
With people's pulmonary carcinoma SPC tumor cell (Chinese Academy of Sciences's Shanghai cell provides), with containing RPMI 1640 culture medium culturings of 10% NBCS, the cell of the trophophase of taking the logarithm, with behind 0.25% trypsinization with 3 * 10
3Individual/hole is inoculated in 96 orifice plates, places 37 ℃, 5%CO
2Cultivated in the incubator 24 hours, and established 8 multiple holes for every group, test group adds five kinds of bases, and each multiple hole drug level of five kinds of bases and matched group 5-FU is 0.5mg/mL, the every hole 200 μ L of culture fluid, the aseptic filtering with microporous membrane of 0.22 μ m.The Measuring Time of drafting 48 hours, inverted microscope observation of cell form was also taken pictures, and every then hole adds the MTT solution 20 μ L of 5mg/mL, places 37 ℃, 5%CO
2Continue in the incubator to cultivate 4 hours, stop cultivating, inhale and abandon the supernatant of cultivating in the hole; Every hole adds 150 μ L DMSO, shakes 10 minutes, crystal is fully dissolved after; Measure 570mn place absorbance with automatic ELIASA (Thermo, model MULTISKAN MK3).Each plate is established 8 multiple holes for every group, each experiment repetition 3 times, and data are the average of three experiments in the table 2.
Growth of tumour cell suppression ratio computing formula is following:
The OD value of the actual OD value/negative control hole in tumor cell survival rate (%)=dosing hole;
Growth of tumour cell suppression ratio (%)=100%-cell survival rate.
Table 2: 48,72 hours suppression ratio (%) of the normal five kinds of base pair people pulmonary carcinoma SPC tumor cells of people
Wherein, A, G, C, T, U represent adenine, guanine, cytosine, thymus pyrimidine and uracil respectively.
Two, result and analysis
From last table 2, can analyze:
1, guanine is because dissolubility is lower, and the part medicine does not dissolve by filtering, actual concentrations 0.3mg/mL;
2, the suppression ratio of adenine is apparently higher than other base, and adenine is exactly adenine phosphate, treated leukopenia.
Embodiment 3
A kind of tumor that presses down of combination is tested in adenine and other four kinds of bases.
One, test method
With people's hepatocarcinoma HepG2 tumor cell (Chinese Academy of Sciences's Shanghai cell provides), with containing RPMI 1640 culture medium culturings of 10% NBCS, the cell of the trophophase of taking the logarithm, with behind 0.25% trypsinization with 3 * 10
3Individual/hole is inoculated in 96 orifice plates, places 37 ℃, 5%CO
2Cultivated 24 hours in the incubator, establish 8 multiple holes for every group, test group adds A+G, A+C, A+T, A+U, and the combination molar ratio of adenine and other 4 kinds of bases is 1:1, and the concentration of two kinds of bases is respectively 0.25mg/mL.Each multiple hole medicine total concentration of 4 kinds of combinations of test group and matched group 5-FU is 0.5mg/mL.The every hole 200 μ L of culture fluid, the aseptic filtering with microporous membrane of 0.22 μ m.The Measuring Time of drafting 48 hours, inverted microscope observation of cell form was also taken pictures, and every then hole adds the MTT solution 20 μ L of 5mg/mL, places 37 ℃, 5%CO
2Continue in the incubator to cultivate 4 hours, stop cultivating, inhale and abandon the supernatant of cultivating in the hole; Every hole adds 150 μ L DMSO, shakes 10 minutes, crystal is fully dissolved after; Measure 570mn place absorbance with automatic ELIASA (Thermo, model MULTISKAN MK3).Each plate is established 8 multiple holes for every group, each experiment repetition 3 times, and data are the average of three experiments in the table 3.
Growth of tumour cell suppression ratio computing formula is following:
The OD value of the actual OD value/negative control hole in tumor cell survival rate (%)=dosing hole;
Growth of tumour cell suppression ratio (%)=100%-cell survival rate.
Table 3: the combination of adenine and other a kind of base is to the suppression ratio (%) of people's hepatocarcinoma HepG2
Wherein, A, G, C, T, U represent adenine, guanine, cytosine, thymus pyrimidine and uracil respectively.
Two, result and analysis
Visible by table 3:
1, after adenine and any one other base mol ratio 1:1 combination, antitumor action slightly descends, but the absolute concentration of medicine descends 1/2;
2, the external tumor-inhibiting action A+T of four kinds of combinations is the strongest, A+C a little less than.
Embodiment 4
The tumor that presses down of the wantonly two kinds of combinations in adenine and other four kinds of bases is tested.
One, test method
With gastric carcinoma cells BGC823 tumor cell (Chinese Academy of Sciences's Shanghai cell provides), with containing RPMI 1640 culture medium culturings of 10% NBCS, the cell of the trophophase of taking the logarithm, with behind 0.25% trypsinization with 3 * 10
3Individual/hole is inoculated in 96 orifice plates, places 37 ℃, 5%CO
2Cultivated 24 hours in the incubator; Establish 8 multiple holes for every group; Test group adds A+C+U, A+G+C, A+C+T, A+T+U, A+U+G; Any two kinds of combination mol ratios of adenine and other four kinds of bases are 1:1:1, and each multiple hole medicine total concentration is 0.5mg/mL, and the concentration of three kinds of bases is respectively 0.167mg/mL.The concentration of matched group 5-FU is 0.5mg/mL.The every hole 200 μ L of culture fluid, the aseptic filtering with microporous membrane of 0.22 μ m.The Measuring Time of drafting 48 hours, inverted microscope observation of cell form was also taken pictures, and every then hole adds the MTT solution 20 μ L of 5mg/mL, places 37 ℃, 5%CO
2Continue in the incubator to cultivate 4 hours, stop cultivating, inhale and abandon the supernatant of cultivating in the hole; Every hole adds 150 μ L DMSO, shakes 10 minutes, crystal is fully dissolved after; Measure 570mn place absorbance with automatic ELIASA (Thermo, model MULTISKAN MK3).Each plate is established 8 multiple holes for every group, each experiment repetition 3 times, and data are the average of three experiments in the table 3.
Growth of tumour cell suppression ratio computing formula is following:
The OD value of the actual OD value/negative control hole in tumor cell survival rate (%)=dosing hole;
Growth of tumour cell suppression ratio (%)=100%-cell survival rate.
Table 4: adenine and other two kinds of base combinations are to gastric carcinoma cells BGC823 suppression ratio (%)
Wherein, A, G, C, T, U represent adenine, guanine, cytosine, thymus pyrimidine and uracil respectively.
Two, result and analysis
Visible by table 4:
1, after adenine and other two kinds other base mol ratio 1:1:1 made up, antitumor action had decline, but the absolute concentration of medicine descends 2/3;
2, the external tumor-inhibiting action A+T+U of five kinds of combinations is the strongest, A+G+C a little less than.
Embodiment 5
The oral capsule agent producing process
1, specification and prescription
1.1 prescription: 10mg/ grain
Adenine: 10g (in anhydride)
Microcrystalline Cellulose: 66g
Carboxymethyl starch sodium: 15g
Magnesium stearate: 2g
Process: 1000.
1.2 each component effect in the prescription
Adenine: principal agent; Microcrystalline Cellulose: diluent; Carboxymethyl starch sodium: disintegrating agent; Magnesium stearate: lubricant.
2, technology
(1) the adenine crushed after being dried is crossed 60 mesh sieves, microcrystalline Cellulose, carboxymethyl starch sodium, 80 ℃ of dryings of magnesium stearate are crossed 100 mesh sieves, and are subsequent use;
(2) take by weighing adenine, microcrystalline Cellulose, carboxymethyl starch sodium, the magnesium stearate of formula ratio, mix homogeneously;
(3) encapsulated;
(4) sampling inspection entirely, pack after qualified finished product.
Two, method for using
The method for using that is used for the people 60-120mg/ days, divides 3 ante cibum oral.
Embodiment 6
Adenine lotus tumor rabbit MRI pharmacodynamic study
Application background inhibition The Diffusion MR Images technology and hydrogen proton magnetic resonance wave spectrum dynamic monitoring adenine are to the treatment afterreaction of VX2 liver transplantation tumor; Inquire into the effect of magnetic resonance molecular imaging in adenine antitumor drug effect is estimated, its Anticancer Effect and Mechanism of preliminary identification in conjunction with relevant pathological change.The leftlobe of liver that celiophyma piece planting method is planted the tumor piece the purebred White Rabbit of New Zealand is opened in employing.Inoculation back is with the size and the growing state of two-dimensional ultrasound monitoring tumor, when treating that tumor is grown to the 0.5-1.5cm size, lotus tumor rabbit is divided into totally three groups of adenine group (A group), Endostatin group (B group) and blank groups (C group), 8 every group at random.Begin the auricular vein injectable drug after 14 days in the lotus tumor, the A group gives adenine 4.0mg/kg; The B group gives Endostatin 1.0mg/kg; C group gives normal saline 0.3mL/kg, medication every day once, continuous use 10 days.Three groups of all 2,7,13 days conventional sequences of capable MR, STIR-EPI-DWI inspections after preceding 1 day of medication and medication, 10 days capable 1H-MRS check after preceding 1 day of medication and the medication.Measure respectively and respectively organize tumor tissues compound choline (choline before and after different time points apparent diffusion coefficient ADC value and treatment; Cho) peak and lipid (lipid; Lip) peak height at peak is measured gross tumor volume simultaneously and is estimated the signal characteristic on T1WI, the T2WI image.The treatment back was put to death the Total Test animal on the 14th day and is taken out the tumor piece; The fresh tumor tissues of the aseptic part of drawing materials is done the expression that fluorescence real-time quantitative PCR detects tumor VEGF-A mRNA, and remaining tumor tissues is got greatest cross-section, and 4% paraformaldehyde is fixed; FFPE, 5 μ m serial section; The HE that goes respectively dyes and carries out pathological examination, and immunohistochemical staining detects the expression of tumor CD31, and the TUNEL method detects the apoptosis of tumor cell.The result shows that the success ratio of inoculation of tumor is 100%, chemotherapy first three groups tumor size do not have statistical significance (
P>0.05).The tumor situation of change sees the following form: table 5: each time point gross tumor volume (cm
3) and relative tumour volume natural logrithm value ln (RTV)
* compare with matched group
P<0.01,
▲Compare with matched group
P<0.05
Table 6:VEGF-A mRNA expresses and distribution and the dependency of MVD counting in each group
* compare with matched group
P<0.01
Table 7: treat tumor tissues ADC value (* 10 after 13 days
-3Mm
2/ s) with the AI dependency
* < 0.01, # and Endostatin group be P < 0.01 relatively to compare P with matched group
Conclusion: 1, adenine has the effect that suppresses rabbit VX2 tumor growth; 2, utilize DWIBS and 1H-MRS technology to estimate the curative effect of adenine in early days dynamically in medication to VX2 liver transplantation tumor; 3, adenine has cell death inducing, suppresses the Anticancer Effect and Mechanism of tumor-blood-vessel growth.
Embodiment 7
One, injection production technology
1, specification and prescription
1.1 prescription: 5mg/mL
Adenine: 50g (in anhydride)
Sodium chloride: 90g
Water for injection: 10000mL
Process 1000 injections.
1.2 each component effect in the prescription
Adenine: principal agent
Sodium chloride: osmotic pressure regulator
Water for injection: solvent.
2, technology
(1) glass ampule is slightly washed earlier the back by the injection requirement and is used water for injection fine purifiation, dry for standby;
(2) in material-compound tank, add injection water 6000mL, add the sodium chloride stirring and dissolving of formula ratio;
(3) add 0.05% needle-use activated carbon, 60 ℃ of stirring and adsorbing 30 minutes, coarse filtration is taken off charcoal; Be heated to and add formula ratio adenine stirring and dissolving more than 80 ℃ again, add to the full amount of water for injection;
(4) stir cooling, fine straining;
(5) sample examination;
(6) be filled in the ampoule sealing by fusing; 115 ℃ of pressure sterilizings 30 minutes.
(7) lamp inspection, packing, warehouse-in.
Two, method for using: 40-100mg/ days, add slowly intravenous drip in 250mL or the 500mL normal saline.
Three, pharmacodynamic study
1. experiment material and method
Laboratory animal: 30 of BALB/c mouses, female ♀, body weight 22 ± 2g is available from Shandong Province's Experimental Animal Center.
Cell strain: mouse junction cancer CT-26 cell strain, be so kind as to give by The 2nd Army Medical College.
Experiment medicine: by the new drug assessment centers preparation of above-mentioned prescription Shandong University.Endostatin: available from Yantai, Shandong Mai Dejin biological engineering limited company.
Method: with the conventional CT-26 cell of cultivating of the RPMI-1640 culture medium that contains 10% calf serum, contain in 37 ℃ and to be cultured to the cell log trophophase in the 5%CO2 constant incubator and to carry out cell inoculation, with 1.5 * 10
6It is subcutaneous that the density 0.2mL of/mL is inoculated in the right side of mice armpit.The lotus tumor after 5 days 30 mices see that all grain of rice size tumor grows, 30 tumor-bearing mices are divided into 3 groups at random, each 0.4mL of armpit subcutaneous injection normal saline, Endostatin (2ug/ days/only), adenine (2mg/ days/only) in the left side respectively in lotus tumor the 5th day.Behind the continuous use 14 days, MR viviperception tumor growth situation, evaluating efficacy.After testing end in 16 days, put to death mice, take by weighing tumor and heavily reach measurement tumor size and carry out pathology and the SABC detection.
2. experimental result
Mouse inoculation is all seen tumor growth after 5 days, lotus tumor success rate 100%.
Table 8: different pharmaceutical is to the inhibitory action of mouse tumor
Conclusion:
1, adenine is obvious to mouse-borne tumor model tumor inhibition effect;
2, the adenine tumor inhibition effect is superior to endostatin.
Embodiment 8
One, release injectable agent producing process
1, specification and prescription
1.1 prescription: 5mg/ props up
Adenine: 50g (in anhydride)
Copolymer of poly lactic acid: 50g
Dichloromethane: 500mL
Sodium carboxymethyl cellulose: 15g
0.9% sodium chloride solution: 10000mL
Process 1000 slow releasing injection.
1.2 each component effect in the prescription
Adenine: principal agent
Copolymer of poly lactic acid: slow-release auxiliary material
Sodium carboxymethyl cellulose: cosolvent
Dichloromethane: solvent
0.9% sodium chloride solution: solvent.
2, production technology
(1) cillin bottle, bottle stopper and aluminium lid are slightly washed earlier the back by the injection requirement and are used water for injection fine purifiation, dry for standby;
(2) copolymer of poly lactic acid with formula ratio adds in the container, adds methylene chloride to dissolve mixing in right amount, adds the adenine mixing of formula ratio again, uses the spray drying method for preparation injectable microsphere;
(3) injectable microsphere that makes is dissolved in the sodium carboxymethyl cellulose normal saline, processes suspension;
(4) suspension is respectively charged into cillin bottle ,-80 ℃ after freezing 24 hours, placed the freeze dryer vacuum drying 48 hours;
(5) sample examination;
(6) tamponade, the jewelling lid;
(7) check, packing, warehouse-in.
Two, method for using
The method for using that is used for the people 40-100mg/ days, adds 250mL normal saline medium-sized vein drop.
Embodiment 9
HT-29 colorectal cancer chick chorioallantoic membrane transplantation model is set up and antineoplastic vascular generates, the research of histiocyte mechanism of proliferation.
One, sets up stable colorectal cancer chick chorioallantoic membrane transplantation model
Observe the dynamic change of chick chorioallantoic membrane vascular development, clearly set up the optimal condition of colorectal cancer chick chorioallantoic membrane transplantation model.The minimum quantity of inoculation HT-29 cell is as the desirable inoculating cell quantity of modelling when selecting the highest tumor formation rate; The HT-29 cell inoculation of identical ideal quantity in the relative avascular area of different days chick chorioallantoic membrane, is confirmed best inoculation embryo age; With the cell inoculation of ideal quantity relative avascular area, understand the tumor bulk-growth and the angiogenesis Changing Pattern of colorectal cancer chick chorioallantoic membrane transplantation model to the righttest age in days chick chorioallantoic membrane.
Two, antitumor mechanism is inquired into
If adenine group, PBS do blank group, the positive matched group of grace degree, the negative matched group of VEGF totally 4 groups.Select to have inoculated the one-tenth tumor Embryo Gallus domesticus of HT-29 cell after 3 days; Becoming the tumor place to place pretreated gelfoam, the experimental group medicine is respectively being got 10 μ l with matched group PBS be added on respectively on each gelfoam of organizing, hatching 5 days; Fixedly get film; Observe different experiments group vascularization characteristic, calculate institute's favored area blood vessel area ratio, and calculate the angiogenesis suppression ratio.Get tumor soma and fix, FFPE, routine paraffin wax section, the HE staining is observed the histologic characteristics of tumor body, adopts immunohistochemistry technology to detect the expression of organizing PCNA PCNA.The result shows that the adenine of 5 μ g/ Embryo Gallus domesticus can suppress the angiogenesis of colorectal cancer chick chorioallantoic membrane transplantation model; It is the propagation through direct inhibition tumor cell that adenine antineoplastic mechanism has more than; But through suppressing the propagation of vascular endothelial cell; And then suppress the tumor tissues vascularization, thereby further suppress tumor growth.
Conclusion:
1, adenine has the effect of obvious inhibition Embryo Gallus domesticus tumor vascular growth;
2, its effect that suppresses the Embryo Gallus domesticus tumor-blood-vessel growth is strong than Endostatin;
3, adenine can suppress tumor new vesselses generations at different levels.
Claims (8)
1. the application of five kinds of normal bases of human body in preparation medicine for treating tumor thing, described five kinds of bases comprise: adenine, guanine, cytosine, thymus pyrimidine and uracil.
2. the application of five kinds of normal bases of human body according to claim 1 in preparation medicine for treating tumor thing, wherein, described base is one or more in adenine, guanine, cytosine, thymus pyrimidine and the uracil.
3. the application of five kinds of normal bases of human body according to claim 1 in preparation medicine for treating tumor thing, wherein, described base is the combination of one or both and adenine in guanine, cytosine, thymus pyrimidine or the uracil.
4. the application of five kinds of normal bases of human body according to claim 1 in preparation medicine for treating tumor thing, wherein, described tumor is pulmonary carcinoma, colon cancer, gastric cancer, the esophageal carcinoma, hepatocarcinoma, cancer of pancreas, breast carcinoma, genital system tumor or glioma.
5. according to the application of five kinds of normal bases of the described human body of claim 1-4 in preparation medicine for treating tumor thing, wherein said medicine is oral tablet, injection, slow releasing agent or targeting preparation.
6. the application of five kinds of normal bases of human body according to claim 5 in preparation medicine for treating tumor thing; The method for preparing of said oral tablet is that described base is added in the adjuvant; The part by weight of base and adjuvant is 1:1, and processing base contents is 10-30mg/ sheet oral tablet.
7. the application of five kinds of normal bases of human body according to claim 5 in preparation medicine for treating tumor thing; The method for preparing of said injection is for directly to be dissolved in described base in the phosphate buffer solution; Process the injection that the base total concentration is 5-25mg/mL, every injection volume is 2-10mL.
8. the application of five kinds of normal bases of human body according to claim 5 in preparation medicine for treating tumor thing; The method for preparing of said slow releasing agent is: medicament liposome, microcapsule or micelle medicine-releasing system are processed slow releasing agent with described base, then slow releasing agent are made injection.
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| CN2011103613136A CN102406649A (en) | 2011-11-15 | 2011-11-15 | Application of five normal basic groups of human body in preparation of medicines for treating tumors |
| PCT/CN2012/001515 WO2013071696A1 (en) | 2011-11-15 | 2012-11-12 | Use of five normal bases in humans for preparation of tumour drugs |
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| WO2013071696A1 (en) * | 2011-11-15 | 2013-05-23 | Zhang Shizhuang | Use of five normal bases in humans for preparation of tumour drugs |
| WO2015042821A1 (en) * | 2013-09-26 | 2015-04-02 | 华安医学股份有限公司 | Compound for activating ampk and uses thereof |
| CN104510736A (en) * | 2013-09-26 | 2015-04-15 | 华安医学股份有限公司 | Compound for activating AMPK and application thereof |
| CN104622887A (en) * | 2014-11-24 | 2015-05-20 | 张始状 | Antitumor drug prepared through combination of purine deoxynucleoside and other nucleosides or basic groups as well as preparation method and applications thereof |
| JP2018058877A (en) * | 2017-11-29 | 2018-04-12 | 華安醫學股分有限公司Ｅｎｅｒｇｅｎｅｓｉｓ Ｂｉｏｍｅｄｉｃａｌ Ｃｏ．， Ｌｔｄ． | Ampk-activating compound, and use thereof |
| JP2018058876A (en) * | 2017-11-29 | 2018-04-12 | 華安醫學股分有限公司Ｅｎｅｒｇｅｎｅｓｉｓ Ｂｉｏｍｅｄｉｃａｌ Ｃｏ．， Ｌｔｄ． | Ampk-activating compound, and use thereof |
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Application publication date: 20120411 |