CN105435215A - Application of recombinant human calcineurin B subunit - Google Patents
Application of recombinant human calcineurin B subunit Download PDFInfo
- Publication number
- CN105435215A CN105435215A CN201511015898.0A CN201511015898A CN105435215A CN 105435215 A CN105435215 A CN 105435215A CN 201511015898 A CN201511015898 A CN 201511015898A CN 105435215 A CN105435215 A CN 105435215A
- Authority
- CN
- China
- Prior art keywords
- administration
- group
- rhcnb
- injection
- dose
- 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
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/43—Enzymes; Proenzymes; Derivatives thereof
- A61K38/46—Hydrolases (3)
- A61K38/465—Hydrolases (3) acting on ester bonds (3.1), e.g. lipases, ribonucleases
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y301/00—Hydrolases acting on ester bonds (3.1)
- C12Y301/03—Phosphoric monoester hydrolases (3.1.3)
- C12Y301/03016—Phosphoprotein phosphatase (3.1.3.16), i.e. calcineurin
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Veterinary Medicine (AREA)
- Medicinal Chemistry (AREA)
- Public Health (AREA)
- Organic Chemistry (AREA)
- Animal Behavior & Ethology (AREA)
- Epidemiology (AREA)
- Pharmacology & Pharmacy (AREA)
- General Engineering & Computer Science (AREA)
- Dermatology (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Biochemistry (AREA)
- Gastroenterology & Hepatology (AREA)
- Immunology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention relates to the field of proteins, in particular to application of recombinant human calcineurin B subunit, and mainly provides application of rhCNB to preparation of a medicine for killing and/or inhibiting a liver cancer cell Bel-7402. After continuous medication for 6 times, medication groups of rhCNB for injection with doses of 10 mg/kg, 20 mg/kg and 40 mg/kg all have a relatively good inhibiting effect on the growth of human liver cancer cell Bel-7402 solid tumors. Subsequent testing results show that the effects of high-dose, middle-dose and increasing-dose medication groups are superior to that of a low-dose medication group, and the treating effect of rhCNB for injection is equivalent to those of recombinant human interleukin-2 for injection and hydroxycamptothecine for injection in a solvent control group. The results of d7 testing after final medication show that the high-dose group has the optimum controlling effect (p is smaller than 0.01) on tumor cell proliferation, which is superior to those of other dose groups and a positive control group, and equivalent to those of a hydroxycamptothecine medication group and the increasing-dose group (p is smaller than 0.05).
Description
Technical field
The present invention relates to albumen field, particularly the application of Recombinant Human Calcineurin B Subunit.
Background technology
Calcineurin (Calcineurin, CN) is that known unique one relies on Ca at present
2+the phosphoprotein phosphatase of/CaM (calmodulin), is made up of with the ratio of 1:1 A, B subunit.A subunit (CNA) is catalytic subunit, and B subunit (CNB) regulates subunit.CN plays a significant role in immune activation path, shows that the key of T cell activation does not have.CN as macromole pheron, easy in inactivation, unstable, and CNB is as the adjustment subunit of this enzyme, can promote the activity of CNA, and relative molecular mass is little, stable in properties.Show that CNB can stimulate the killing activity of T cell and NK cell proliferation and enhancing NK cell by research, strengthen the immunologic functions such as macrophages phagocytic capacity, and Recombinant Human Calcineurin B Subunit (RecombinanthumanCalcineurinBsubunit, rhCNB) is the genetic engineering innovation tumour medicine in exploitation.
But have no rhCNB at present to kill and wound hepatoma carcinoma cell Bel-7402 or inhibiting relevant report.
Summary of the invention
In view of this, the invention provides the application of Recombinant Human Calcineurin B Subunit.The present invention adopts IVISKinetics small animal living body imaging system, injection rhCNB is to the therapeutical effect of BALB/c nude mouse lotus Human hepatoma cell line Bel-7402 cancer in situ in research, further investigation tested material injection rhCNB killing and wounding and (or) inhibitory action particular type tumor cell, explore timeliness and dose-effect relationship that tested material produces drug action, for effectiveness and other toxicology correlation test of medicine provide foundation.
In order to realize foregoing invention object, the invention provides following technical scheme:
The invention provides rhCNB kill and wound in preparation and/or suppress the application in the medicine of hepatoma carcinoma cell Bel-7402.
In specific embodiments more of the present invention, rhCNB kills and wounds in preparation and/or suppresses the dosage of rhCNB described in the application in the medicine of hepatoma carcinoma cell Bel-7402 for being no less than 10mg/kg.
In specific embodiments more of the present invention, rhCNB kills and wounds in preparation and/or suppresses the dosage of rhCNB described in the application in the medicine of hepatoma carcinoma cell Bel-7402 to be 10mg/kg ~ 160mg/kg.
In specific embodiments more of the present invention, rhCNB kills and wounds in preparation and/or suppresses the administration concentration of rhCNB described in the application in the medicine of hepatoma carcinoma cell Bel-7402 to be 0.5mg/mL ~ 2mg/mL.
In specific embodiments more of the present invention, rhCNB kills and wounds in preparation and/or suppresses the administering mode of rhCNB described in the application in the medicine of hepatoma carcinoma cell Bel-7402 to be intravenous administration; Daily 1 time, successive administration 15 days or first administration dosage are 10mg/kg, within every 3 days, improve 1 multiple dose and increase progressively administration, be followed successively by 10mg/kg, 20mg/kg, 40mg/kg, 80mg/kg, 160mg/kg.
Present invention also offers the application of rhCNB in the medicine of preparation treatment BALB/c nude mouse lotus Human hepatoma cell line Bel-7402 cancer in situ transplanted tumor.
In specific embodiments more of the present invention, described in the application of rhCNB in the medicine of preparation treatment BALB/c nude mouse lotus Human hepatoma cell line Bel-7402 cancer in situ transplanted tumor, the dosage of rhCNB is for being no less than 10mg/kg.
In specific embodiments more of the present invention, described in the application of rhCNB in the medicine of preparation treatment BALB/c nude mouse lotus Human hepatoma cell line Bel-7402 cancer in situ transplanted tumor, the dosage of rhCNB is 10mg/kg ~ 160mg/kg.
In specific embodiments more of the present invention, described in the application of rhCNB in the medicine of preparation treatment BALB/c nude mouse lotus Human hepatoma cell line Bel-7402 cancer in situ transplanted tumor, the administration concentration of rhCNB is 0.5mg/mL ~ 2mg/mL.
In specific embodiments more of the present invention, described in the application of rhCNB in the medicine of preparation treatment BALB/c nude mouse lotus Human hepatoma cell line Bel-7402 cancer in situ transplanted tumor, the administering mode of rhCNB is intravenous administration; Daily 1 time, successive administration 15 days or first administration dosage are 10mg/kg, within every 3 days, improve 1 multiple dose and increase progressively administration, be followed successively by 10mg/kg, 20mg/kg, 40mg/kg, 80mg/kg, 160mg/kg.
At experimental session, except positive control cisplatin for inj high dose administration treated animal, the general observation of symptoms of all the other animals and body weight compare no significant difference with Vehicle controls group.In successive administration 6 ~ 15 processes, each dosed administration of injection rhCNB more all has statistical significance in various degree to the control of tumor cell proliferation and Vehicle controls group.
Testing result display on the 7th after stopping administration: compare with Vehicle controls group, G administration group all has remarkable statistical significance to the control of tumor cell proliferation, p < 0.01, DZ, X administration group have statistical significance, p < 0.05.
After successive administration 6 times, injection rhCNB10mg/kg, 20mg/kg, 40mg/kg dosed administration group all have good inhibitory action to the growth of Human hepatoma cell line Bel-7402 solid tumor.Testing result display in administration process after this, high, in and ascending-dose group effect be better than low dosage administration group, therapeutic effect and injection RhIL-2, hydroxycamptothecin positive control are suitable.After last administration, d7 testing result shows, high dose group is to control effects the best (p < 0.01) of tumor cell proliferation, be better than other dosage group and positive controls, hydroxycamptothecin administration group and ascending-dose group are to the control effects of tumor cell proliferation quite (p < 0.05).
Accompanying drawing explanation
In order to be illustrated more clearly in the embodiment of the present invention or technical scheme of the prior art, be briefly described to the accompanying drawing used required in embodiment or description of the prior art below.
Fig. 1 shows experiment each group of knubble biological luminous detection data
wherein abscissa " 1 ~ 7 " represents " before administration, after first administration after d3, d6, d9, d12, d15 and last administration convalescent period d7 " respectively; " G-injection rhCNB high dose group (40mg/kg) " " in Z-injection rhCNB dosage group (20mg/kg) ", " D-injection rhCNB low dose group (10mg/kg) ", DZ-injection rhCNB dosage escalation group (10mg/kg → 20mg/kg → 40mg/kg → 80mg/kg → 160mg/kg, 10mg/kg is predose, successive administration 3d improves a dosage, by that analogy) "; Vertical coordinate is bioluminescent detection intensity;
Fig. 2 shows living imaging figure;
Fig. 3 shows that experimental group cuts open inspection figure;
Fig. 4 shows that positive controls cuts open inspection figure.
Detailed description of the invention
The invention discloses the application of Recombinant Human Calcineurin B Subunit, those skilled in the art can use for reference present disclosure, and suitable improving technique parameter realizes.Special needs to be pointed out is, all similar replacements and change apparent to those skilled in the art, they are all deemed to be included in the present invention.Method of the present invention and application are described by preferred embodiment, related personnel obviously can not depart from content of the present invention, spirit and scope methods and applications as herein described are changed or suitably change with combination, realize and apply the technology of the present invention.
The present invention is according to " cell toxicant series antineoplastic medicament non-clinical study technological guidance principle " and " antitumor drug pharmacodynamics guideline ", adopt IVISKinetics small animal living body imaging system, dynamic analysis injection rhCNB is to the therapeutical effect of BALB/c nude mouse lotus Human hepatoma cell line Bel-7402 cancer in situ.First set up BALB/c nude mouse lotus Human hepatoma cell line Bel-7402 subcutaneous transplantation tumor model, then set up BALB/c nude mouse lotus Human hepatoma cell line Bel-7402 Orthotopic Transplantation Model.Get the successful nude mouse of modeling 90, be divided into 9 administration groups by bioluminescence digital average, comprise: injection rhCNB high dose group (dosage is 40mg/kg, and administration concentration is 2mg/mL), dosage group (dosage is 20mg/kg, and administration concentration is 1mg/mL) in injection rhCNB, injection rhCNB low dose group (dosage is 10mg/kg, and administration concentration is 0.5mg/mL), (first administration dosage is 10mg/kg to injection rhCNB dosage escalation group, within every 3 days, improves 1 multiple dose, is followed successively by 10mg/kg, 20mg/kg, 40mg/kg, 80mg/kg, 160mg/kg), positive control injection RhIL-2 administration group (dosage is 16.4 ten thousand IU/kg, and administration concentration is 8200IU/mL), positive control cisplatin for inj high dose administration group (5mg/kg, administration concentration is 0.25mg/mL), positive control cisplatin for inj low dosage administration group (1mg/kg, administration concentration is 0.05mg/mL), positive control injection hydroxycamptothecin administration group (2mg/kg, administration concentration is 0.1mg/mL) and Vehicle controls group.Test each group daily 1 time, except positive control cisplatin for inj high dose administration group successive administration 5 days, all the other respectively organize successive administration 15.Before each administration and after administration, general observation of symptoms is carried out to animal.Within before first administration after (same day), first administration the 7th day behind 3 days and last administration, detect 1 time by CaliperIVISKinetics small animal living body imaging system respectively, before each detection, the weight of animals is weighed.Last detects and terminates rear execution animal, substantially cuts open inspection, takes out tumor inoculation organ (comprising tumor tissues), carries out pathologic finding and the detection of apoptotic SABC.
Result is at experimental session, and except positive control cisplatin for inj high dose administration treated animal, the general observation of symptoms of all the other animals and body weight compare no significant difference with Vehicle controls group.In successive administration 6 ~ 15 processes, each dosed administration of injection rhCNB more all has statistical significance in various degree to the control of tumor cell proliferation and Vehicle controls group.
Testing result display on the 7th after stopping administration: compare with Vehicle controls group, G administration group all has remarkable statistical significance to the control of tumor cell proliferation, p < 0.01, DZ, X administration group have statistical significance, p < 0.05.
After conclusion successive administration 6 times, injection rhCNB10mg/kg, 20mg/kg, 40mg/kg dosed administration group all have good inhibitory action to the growth of Human hepatoma cell line Bel-7402 solid tumor.Testing result display in administration process after this, high, in and ascending-dose group effect be better than low dosage administration group, therapeutic effect and injection RhIL-2, hydroxycamptothecin positive control are suitable.After last administration, d7 testing result shows, high dose group is to control effects the best (p < 0.01) of tumor cell proliferation, be better than other dosage group and positive controls, hydroxycamptothecin administration group and ascending-dose group are to the control effects of tumor cell proliferation quite (p < 0.05).
The invention provides raw materials used in the application of Recombinant Human Calcineurin B Subunit and reagent all to be buied by market.
Tumor cell line: the hepatoma cell strain Bel-7402 of luc bioluminescence marker is purchased from Le Chen bio tech ltd, Shanghai.
Cell culture fluid: gibcoRPMI1640 cell culture fluid (500mL/ bottle)
Hyclone: MP hyclone (500mL/ bottle)
Cell dissociation buffer: gibco cell dissociation buffer
Substrate: Caliper Lampyridea D-fluorescein potassium salt
Anesthetis: isoflurane
Laboratory animal: BALB/c nude mouse, is purchased from Guangdong Medical Lab Animal Center, the quality certification is numbered № 44007200012061.
Below in conjunction with embodiment, set forth the present invention further:
The foundation of embodiment 1BALB/c nude mouse lotus Human hepatoma cell line Bel-7402 subcutaneous transplantation tumor model
The external cellar culture of Human hepatoma cell line Bel-7402, digest centrifugal after make concentration and be about 1 × 10
7the cell suspension of individual cell/mL, it is subcutaneous that 0.2mL/ is only inoculated in right side of mice axillary fossa.
The foundation of embodiment 2BALB/c nude mouse lotus Human hepatoma cell line Bel-7402 Orthotopic Transplantation Model
Take out well-grown subcutaneous transplantation tumor, the tumor tissue of being cut into about 2 × 2 × 1mm size is for subsequent use.With isoflurane, gas anesthesia is carried out to animal to be seeded, preoperative feed 12h.Get dorsal position after Animal Anesthesia, under xiphoid-process, 2 ~ 5mm place laterally successively cuts off skin, and opening is about 1cm, exposes liver, and inoculated layer-by-layer suture skin after tumor, An Erdian disappears, and puts back to normal in cage raising.Post operation 7d, is undertaken detecting, dividing into groups and administration by small animal living body imaging system.
Embodiment 3 animal divides into groups
Get the nude mouse 90 inoculated, be divided into dosage group in injection rhCNB high dose group, injection rhCNB, injection rhCNB low dose group, injection rhCNB dosage escalation group, positive control injection RhIL-2 administration group, positive control cisplatin for inj high dose administration group, positive control cisplatin for inj low dosage administration group, positive control injection hydroxycamptothecin administration group and Vehicle controls group 9 groups at random by bioluminescent detection data, often organize 10.
Embodiment 4 dose design
1.2.4.1 test sample injection rhCNB dose design
" high dose is no more than the maximum tolerated dose of tested material " with reference to " treatment group establishes high, medium and low dosage treatment group; generally press 4:2:1 and arrange " in " antitumor drug pharmacodynamics guideline ", in " cell toxicant series antineoplastic medicament non-clinical study technological guidance principle ", intend clinical first course for the treatment of consumption be 60mg/ day and clinical second, third, the 4th course for the treatment of intended employing ascending-dose.Therefore, test sample injection rhCNB administration group establishes dosage group in test sample high dose group, test sample, test sample low dose group and test sample dosage escalation group 4 groups altogether, and dosage and concentration are in table 1.
1.2.4.2 the dose design of positive drug
1.2.4.2.1 the dose design of positive drug injection RhIL-2
Be according to conventional dosage clinical in injection RhIL-2 description 10 ~ 800,000 IU/ time, daily 1 time.Be that 60kg is calculated as 800,000 IU ÷ 60 ≈ 1.33 ten thousand IU/kg by human body average weight, being scaled mice dose,equivalent is 1.33 ten thousand IU/kg × 12.33 ≈ 16.4 ten thousand IU/kg, and administration concentration is 0.82 ten thousand IU/mL.
1.2.4.2.2 the dose design of positive drug cisplatin for inj
According to illustrating in cisplatin for inj description that general clinical day dosage is 15-20mg/m2 body surface area, successive administration 5d weekly.Human body average body surface area is 1.62m
2, namely clinical administration dosage is 15mg/m2 × 1.62m
2=24.3mg, be that 60kg is calculated as 24.3mg ÷ 60=0.405mg/kg by human body average weight, being scaled mice dose,equivalent is 0.405mg/kg × 12.33 ≈ 5mg/kg, and as high dose, administration concentration is 0.25mg/mL.Because administration time is 15d successive administration, may there is death in animal, and therefore, more separately establish a low dose group, dosage is 1mg/kg, and administration concentration is 0.05mg/mL.
1.2.4.2.3 the dose design of positive drug injection hydroxycamptothecin
According to illustrating in injection hydroxycamptothecin description that general clinical day dosage is 4-6mg/ time, 1 times/day, 15-30d was 1 course for the treatment of.Be that 60kg is calculated as 6mg ÷ 60=0.1mg/kg by human body average weight, being scaled mice dose,equivalent is 0.1mg/kg × 12.33 ≈ 1.23mg/kg, and this experiment is set to 2mg/kg, and administration concentration is 0.1mg/mL.
Table 1 tests each group of dosage and administration schedule
Embodiment 5
1.2.5 route of administration
Route of administration is tail vein injection.
1.2.6 administration volume: 20mL/kg
1.2.7 administration time/frequency
Test sample injection rhCNB administration administration time and frequency design according to clinical plan time and frequency, and positive controls administration time and frequency design according to clinical application time and frequency.
High, medium and low and the dosage escalation group of test sample all daily 1 time, successive administration 15d;
Vehicle controls group daily 1 time, successive administration 15d;
Positive drug injection interleukin-2 administration group daily 1 time, successive administration 15d;
Positive drug cisplatin for inj high dose administration group daily 1 time, successive administration 5d;
Positive drug cisplatin for inj low dosage administration group daily 1 time, successive administration 15d;
Positive drug injection hydroxycamptothecin administration group daily 1 time, successive administration 15d;
1.2.8 Testing index
1.2.8.1 observation of symptoms
Observe the general symptom of animal before administration and after administration, observation index mainly comprises: whether feed is normal, whether drinking-water is normal, whether feces outward appearance is normal, whether urine outward appearance is normal, whether daily routines are normal, whether breathe normal, whether heart beating is normal, whether eye muzzle has secretions and body weight etc.1.2.8.2 body weight and bioluminescent detection
Within before first administration after (same day), first administration the 7th day behind 3 days and last administration, detect 1 time by CaliperIVISKinetics small animal living body imaging system respectively, before each detection, the weight of animals is weighed, carry out bioluminescent detection by 10 ~ 15min after 0.1mL/10g body weight lumbar injection substrate solution (concentration is 15mg/mL) injection.
1.2.8.3 statistical analysis
Experiment the data obtained SPSS analysis software carries out statistical analysis, compares the significance of difference between administration group and Vehicle controls group and between each administration group.
Embodiment 6 experimental result
2.1 general observation of symptoms
Before administration and after administration, the general symptom of animal is observed; except positive control cisplatin for inj high dose administration group, the symptom such as feed, drinking-water, feces outward appearance, urine outward appearance, daily routines, breathing, heart beating, eye muzzle secretions of all the other each administration treated animal animals compares with Vehicle controls group and has no notable difference.
2.2 body weight
Within before first administration after (same day), first administration the 7th day behind 3 days and last administration, respectively measured body weight is carried out to animal, exist except notable difference (p < 0.05) except positive control cisplatin for inj high dose administration treated animal body weight compares with Vehicle controls group, all the other each administration group body weight compare with Vehicle controls group and have no notable difference, p > 0.05.
2.3 bioluminescent detection
Before first administration after (same day), first administration behind 3 days and last administration the 7th day, carry out bioluminescent detection respectively by CaliperIVIS small animal living body imaging system.Testing result is shown in Fig. 1, Fig. 2 and table 2, cuts open inspection and the results are shown in Figure 3.
According to Fig. 1, following result can be drawn:
D3 testing result display after first administration: obviously, wherein SG administration group propagation is the most obvious for G, SG, SD and Vehicle controls group tumor proliferation.From detection data, tumor cell quantity and the positive control cisplatin for inj administration group of high dose group are similar, growth rate is not significantly suppressed, this may, with body inner recipient in conjunction with relevant, also need to carry out comprehensive descision in conjunction with other immunological testing and the study on mechanism etc. in later stage.
D6 testing result display after first administration: only have Vehicle controls group tumor proliferation the most obvious, " G, DZ, SG, SD, B " respectively organizes and after first administration, d3 testing result is obvious downward trend, and respectively group is almost consistent with last test result for all the other;
D9 testing result display after first administration: compared with a front testing result, SG administration group and Vehicle controls group are sharp increase, and D administration group tumor cell quantity reduces to some extent, respectively group propagation is not obvious for all the other;
D12, d15 testing result display after first administration: except SD administration group tumor inner cell propagation reduces to some extent, all the other respectively organize tumor cell number all slightly increases; D7 testing result display after last administration: sharp increase appears in Vehicle controls group tumor cell number amount, and Z, B, D, SD administration group tumor cell quantity also increases to some extent, and G, ZD, X administration group tumor cell quantity has to be increased slightly.
These results suggest that: SG administration group tumor cell quantity presents and repeatedly rebounds during administration, G, D, DZ administration group is not so good as middle Z administration group to the control effects of tumor cell quantity after first administration d3.In administration process after this, except Vehicle controls group and SG administration group, all the other tumor cell quantity growth trends respectively organized are also not obvious, compare, all serve good control effects to the propagation of tumor cell with Vehicle controls group.During last administration d7, test each group of tumor cell quantity all in increase in various degree, but compare propagation trend with Vehicle controls group and not obvious.
Table 2 tests p value and the significance of difference analysis in each group of different dosing period
Note: the number in bracket " () " is surviving animals number; G-injection rhCNB high dose administration group, dosed administration group in Z-injection rhCNB, D-injection rhCNB low dosage administration group, DZ-injection rhCNB dosage escalation administration group, SG-positive control drug cisplatin for inj high dose administration group SD-positive control drug cisplatin for inj low dosage administration group, B-positive control drug injection interleukin-2 administration group, X-positive control drug injection hydroxycamptothecin administration group, Y-Vehicle controls group; Before d0-administration, d7 after d15, d21-last administration after d12, d15-first administration after d9, d12-first administration after d6, d9-first administration after d3, d6-first administration after d3-first administration; "
*" p < 0.05, "
*" p < 0.01, "
Δ" p < 0.001.
Table 2 result shows:
After successive administration 3 times, each administration group compares not statistically significant with Vehicle controls group, the equal > 0.05 of p value.
Testing result display after successive administration 6 times: the tumor cell proliferation situation of injection rhCNB each dosed administration group and positive control administration group all obtains good control, but compare not statistically significant with Vehicle controls group, the equal > 0.05 of p value.
Testing result display after successive administration 9d: injection rhCNBG, Z, D, DZ administration group and positive control B, X administration group more all have extremely significantly statistical significance to the control of tumor cell proliferation and Vehicle controls group, p < 0.001, positive control SD administration group more all has statistical significance with Vehicle controls group, p < 0.05.
Testing result display after successive administration d12: compare with Vehicle controls group, the control of DZ administration group to tumor cell proliferation has extremely significantly statistical significance, p < 0.001, G, Z, D administration group and positive control B, X administration group all have remarkable statistical significance, p < 0.01, positive control SD administration group more all has statistical significance with Vehicle controls group, p < 0.05.
Testing result display after successive administration d15: compare with Vehicle controls group, G, Z, DZ, SD, B, X administration group all has remarkable statistical significance to the control of tumor cell proliferation, p < 0.01, D administration group has statistical significance, p < 0.05.
D7 testing result display after last administration: compare with Vehicle controls group, G administration group all has remarkable statistical significance to the control of tumor cell proliferation, p < 0.01, DZ, X administration group have statistical significance, p < 0.05.
3 conclusions
At injection rhCNB in the therapeutical effect of BALB/c nude mouse lotus Human hepatoma cell line Bel-7402 cancer in situ transplanted tumor, after successive administration 9 times, injection rhCNB10mg/kg, 20mg/kg, 40mg/kg dosed administration group and dosage escalation group all have very effective inhibitory action to the growth of Human hepatoma cell line Bel-7402 solid tumor.Testing result display in administration process after this, high, in and ascending-dose group effect be better than low dosage administration group, effect and injection RhIL-2, hydroxycamptothecin positive control are suitable.After last administration, d7 testing result shows, high dose group is to control effects the best (p < 0.01) of tumor cell proliferation, be better than other dosage group and positive controls, hydroxycamptothecin administration group and ascending-dose group are to the control effects of tumor cell proliferation quite (p < 0.05).After drug withdrawal 7d, injection rhCNB is except high dose group, and the propagation of all the other each dosage groups and positive controls tumor cell there will be raising in various degree.
Comprehensive above analysis, can tentatively draw the following conclusions:
At injection rhCNB in the therapeutical effect of BALB/c nude mouse lotus Human hepatoma cell line Bel-7402 cancer in situ transplanted tumor, effective treatment of tumor (is compared with Vehicle controls group, p < 0.05) time is after successive administration 6 times, during whole administration, tumor can be controlled effectively.
Drug withdrawal is after 1 week, and tumor there will be propagation in various degree, and this and positive controls are similar.But the therapeutic effect that injection rhCNB high dose group and dosage escalation group can keep Tumor suppression to grow, compares with Vehicle controls group, and the p value < 0.01 of high dose group, has remarkable statistical significance; The p value < 0.05 of dosage escalation group, has statistical significance; Positive controls only has the p value < 0.05 of injection hydroxycamptothecin administration group, has statistical significance; The p value > 0.05 of cisplatin for inj low dosage administration group and injection RhIL-2 administration group, not statistically significant.In general, in injection rhCNB, low dose group is suitable to the therapeutic effect of tumor and injection RhIL-2, is better than cisplatin for inj; Injection rhCNB high dose group and ascending-dose group, to therapeutic effect the best of tumor, are better than other dosage group and all positive controls.
The above is only the preferred embodiment of the present invention; it should be pointed out that for those skilled in the art, under the premise without departing from the principles of the invention; can also make some improvements and modifications, these improvements and modifications also should be considered as protection scope of the present invention.
Claims (10)
1.rhCNB kills and wounds in preparation and/or suppresses the application in the medicine of hepatoma carcinoma cell Bel-7402.
2. application according to claim 1, is characterized in that, the dosage of described rhCNB is for being no less than 10mg/kg.
3. application according to claim 1 and 2, is characterized in that, the dosage of described rhCNB is 10mg/kg ~ 160mg/kg.
4. the application according to any one of claims 1 to 3, is characterized in that, the administration concentration of described rhCNB is 0.5mg/mL ~ 2mg/mL.
5. the application according to any one of Claims 1-4, is characterized in that, the administering mode of described rhCNB is intravenous administration; Daily 1 time, successive administration 15 days or first administration dosage are 10mg/kg, within every 3 days, improve 1 multiple dose and increase progressively administration, be followed successively by 10mg/kg, 20mg/kg, 40mg/kg, 80mg/kg, 160mg/kg.
The application of 6.rhCNB in the medicine of preparation treatment BALB/c nude mouse lotus Human hepatoma cell line Bel-7402 cancer in situ transplanted tumor.
7. application according to claim 6, is characterized in that, the dosage of described rhCNB is for being no less than 10mg/kg.
8. the application according to claim 6 or 7, is characterized in that, the dosage of described rhCNB is 10mg/kg ~ 160mg/kg.
9. the application according to any one of claim 6 to 8, is characterized in that, the administration concentration of described rhCNB is 0.5mg/mL ~ 2mg/mL.
10. the application according to any one of claim 6 to 9, is characterized in that, the administering mode of described rhCNB is intravenous administration; Daily 1 time, successive administration 15 days or first administration dosage are 10mg/kg, within every 3 days, improve 1 multiple dose and increase progressively administration, be followed successively by 10mg/kg, 20mg/kg, 40mg/kg, 80mg/kg, 160mg/kg.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201511015898.0A CN105435215A (en) | 2015-12-30 | 2015-12-30 | Application of recombinant human calcineurin B subunit |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201511015898.0A CN105435215A (en) | 2015-12-30 | 2015-12-30 | Application of recombinant human calcineurin B subunit |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN105435215A true CN105435215A (en) | 2016-03-30 |
Family
ID=55546135
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201511015898.0A Pending CN105435215A (en) | 2015-12-30 | 2015-12-30 | Application of recombinant human calcineurin B subunit |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105435215A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108732354A (en) * | 2017-04-14 | 2018-11-02 | 上海市肿瘤研究所 | Applications of the RCAN1.4 as the diagnosis marker of diagnosing hepatocellular carcinoma |
| CN108926713A (en) * | 2017-05-26 | 2018-12-04 | 上海市肿瘤研究所 | The application of calcineurin regulatory protein 1.4 or its analog in the drug that preparation inhibits liver cancer |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1245720A (en) * | 1998-08-26 | 2000-03-01 | 北京师范大学 | Medicinal composition containing calcineurin B subunit |
-
2015
- 2015-12-30 CN CN201511015898.0A patent/CN105435215A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1245720A (en) * | 1998-08-26 | 2000-03-01 | 北京师范大学 | Medicinal composition containing calcineurin B subunit |
Non-Patent Citations (2)
| Title |
|---|
| WEIQ,L IAN ML, J ING FZ等: "《Studies of Calcineurin B Subunit From Genetic Engineering for Use in Medicine》", 《DRUG DEVELOPMENT RESEARCH》 * |
| 高锦等: "《注射用重组人钙调磷酸酶B亚基稳定性的初步研究》", 《中国新药杂志》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108732354A (en) * | 2017-04-14 | 2018-11-02 | 上海市肿瘤研究所 | Applications of the RCAN1.4 as the diagnosis marker of diagnosing hepatocellular carcinoma |
| CN108926713A (en) * | 2017-05-26 | 2018-12-04 | 上海市肿瘤研究所 | The application of calcineurin regulatory protein 1.4 or its analog in the drug that preparation inhibits liver cancer |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105983097B (en) | Anti-tumor preparation and preparation method thereof | |
| CN101896190A (en) | Treatment of melanoma with alpha thymosin peptides in combination with antibodies against cytotoxic t lymphocyte-associated antigen 4 (CTLA4) | |
| CN104159596B (en) | Administration for the antagonist of α 5 β 1 of angiogenesis inhibitor and treatment of cancer | |
| CN105435215A (en) | Application of recombinant human calcineurin B subunit | |
| Shen et al. | Bruton’s tyrosine kinase inhibitors in the treatment of primary central nervous system lymphoma: A mini-review | |
| CN108514639A (en) | It is a kind of to be used for antitumor combination medicine and its preparation and application | |
| Shen et al. | The inhibition of growth and angiogenesis in heterotransplanted esophageal carcinoma via intratumoral injection of arsenic trioxide | |
| CN105435216B (en) | The application of Recombinant Human Calcineurin B Subunit | |
| US20220323450A1 (en) | Methods and compositions for treating neuroblastoma in a juvenile mammalian body | |
| US11541061B2 (en) | Neuroblastoma treatment with taurolidine hydrolysis products | |
| EP1763343B1 (en) | Calcium trifluoroacetate for preparing antiangiogenetic medicaments | |
| EP2981279B1 (en) | Marcks inhibitor peptides for inhibiting metastasis | |
| TWI786186B (en) | Medicinal composition including monoclonal antibody or antigen-binding fragment and use of the same | |
| WO2021155479A1 (en) | Methods for enhancing t cells using venetoclax | |
| CN113117088A (en) | Use of calcium-activated chloride channel inhibitors in tumor immunotherapy | |
| CN112618569A (en) | Medicine for treating urothelial cancer | |
| JP2021535163A (en) | Treatment of neuroblastoma with taurolidine hydrolyzate | |
| CN111249274A (en) | Application of ginkgolide B in preparation of glioma cell activity inhibitor | |
| Gomez-Flores et al. | Increased survival of tumor-bearing mice by the delta opioid SNC 80 | |
| CN111973745B (en) | Application of BAP18 in resisting endocrine treatment of breast cancer | |
| WO2023029141A1 (en) | Use of octenidine dihydrochloride in preparation of antitumor drug | |
| JP2017002043A (en) | Medicine for treating or preventing nerve injury | |
| CN107536845A (en) | A kind of medicine of anti-curing oncoma and application thereof | |
| WO2009138094A1 (en) | Pharmaceutical compositions | |
| CN105250257A (en) | Application of glycine and derivative thereof to preparation of medicine for treating IDH (IsocitrateDehydrogenase) and FH (Fumarate Hydratase) gene mutation related tumors |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20160330 |