CN103505727A - Application of novel function of CD146 targeted as co-receptor of vascular endothelial growth factor receptor-2 (VEGFR-2) in anti-tumor angiogenesis treatment - Google Patents
Application of novel function of CD146 targeted as co-receptor of vascular endothelial growth factor receptor-2 (VEGFR-2) in anti-tumor angiogenesis treatment Download PDFInfo
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Abstract
The invention discloses application of a novel function of a CD146 targeted as a co-receptor of a vascular endothelial growth factor receptor-2 (VEGFR-2) in anti-tumor angiogenesis treatment. In the invention, the condition that a cell adhesion molecule CD146 is the co-receptor of the VEGFR-2 is proposed for the first time, and combined application of an anti-CD146 antibody and an anti-VEGF antibody for inhibition of tumor angiogenesis can become a new strategy for clinical treatment of cancers. Compared with the conventional single molecule-targeted antibody drug for tumor treatment, tumor growth can be more effectively inhibited by combination of two different molecule-targeted antibodies; the CD146 can assist the co-receptor to more effectively transmit a VEGF-mediated downstream signal as the co-receptor of the VEGFR-2, so as to achieve an angiogenesis function; an action mechanism of combination of the anti-CD146 antibody and the anti-VEGF antibody for tumor treatment manly comprises the functions, such as common inhibition of activation of a VEGF-VEGFR-2 signal channel, migration of endothelial cells, angiogenesis and the like by combination of the two antibodies, so as to more effectively inhibit humor growth (such as a pancreatic cancer and the like). Combined application of the two antibodies or cooperation with other clinical treatment means (for example, clinical chemoradiotherapy) is bound to become a new strategy for inhibition of tumor angiogenesis; meanwhile, the CD146 can be applied to basic researches of a tumor angiogenesis mechanism and the like.
Description
Technical field
The present invention relates to be used for the treatment of the compositions of tumor.Particularly, the present invention relates to the compositions of anti-CD146 antibody and VEGF antibody, it can be used for treating tumor.More specifically, the monoclonal antibody bevacizumab of the monoclonal antibody AA98 of the anti-CD146 the present invention relates to or its function fragment and anti-VEGF or the compositions of its function fragment can suppress the activation of VEGF-VEGFR-2 signal path and the function such as the migration of the endotheliocyte that causes and angiogenesis jointly, thereby more effectively suppress tumor growth, more specifically suppress pancreas tumor.
Background technology
Cancer is the No.1 killer of current harm humans life and health.The number of the infected of the annual malignant tumor of China approximately 2,600,000.Die from cancer person and account for the more than 20% of total toll, occupy first (according to statistics in 2005) of urbanite's cause of death.
At present, in the large routine<radiotherapy of tumor three, chemotherapy and operation>in treatment, Drug therapy is an importance.The factor that affects at present tumor pharmacother effect is many-sided, comprises specificity, means of transportation, permeability and the induced tumor drug resistance of medicine.The specificity of its Chinese medicine and drug resistance are that tumor therapeutics is badly in need of the key issue solving.Tumor vasculature targeting has become a kind of important means in oncotherapy with advantages such as its tumour-specific, broad spectrum activity, no or low drug resistance.Its theoretical foundation is mainly the concept of the angiogenesis that the seventies Folkman proposes in last century, and new vessels provides nutrition and oxygen supply for tumor, and then promotes growth and the transfer of tumor.
Large quantity research is found, plant to the factor of closing existing more than 30 with tumor-blood-vessel growth, as vascular endothelial cell growth factor (vascular endothelial growth factor, VEGF), transforming growth factor (transforming growth factor-, TNF), fibroblast growth factor (fibroblast growth factor, FGF), tumor necrosis factor (tumor necrosis factor, TNF), angiogenesis hormone (angiopoietins) etc.Wherein, VEGF is proved to be the topmost somatomedin that directly acts on the high degree of specificity of angiogenesis.Tumor cell can synthesize and the secreted VEGF of periodicity secretion of VEGF combines with the vegf receptor (VEGFRs) on vascular endothelial cell, thereby promotes tumor-blood-vessel growth.The signal transduction of VEGF Angiogensis mainly combination by itself and VEGFR-2 is realized.The key signal molecule that the anti-angiogenic medicaments of listing is all usingd in these signal transduction pathways mostly at present designs as drug target, as the monoclonal antibody bevacizumab (Bevacizumab of anti-VEGF, Roche company), monoclonal antibody DC101 of anti-VEGFR-2 etc.Meanwhile, research finds that VEGFR-2 exists co-receptor, CD44 for example, and it can interact with VEGFR-2, regulates the angiogenesis of VEGFR-2 mediation as its co-receptor.Therefore, this type of co-receptor of targeting also can effectively suppress the angiogenesis of tumor and then suppress tumor growth.
Meanwhile, in the treatment of the antibody mediated immunity of target tumor blood vessel, still have some problems and bottleneck, as Antybody therapy often can not thoroughly kill whole tumor cells, remaining cell easily recurs after drug withdrawal.And the Antybody therapy of single target spot has limitation, can not reach therapeutic effect effectively.Therefore find new treatment target and find that more effective therapeutic strategy is problem demanding prompt solution.
Summary of the invention
The present invention proposes the co-receptor that CD146 is VEGFR-2 first, and the compositions of the monoclonal antibody bevacizumab of the monoclonal antibody AA98 of anti-CD146 and anti-VEGF (Bevacizumab, Roche company) can be used as the novel targeted medicine for the treatment of tumor.In animal tumor model (as human pancreas cancer), to compare with single a kind of antibody drug treatment tumor, the compositions of AA98 and bevacizumab can more effectively suppress tumor-blood-vessel growth and then suppress tumor growth.
The antibody combined application in antineoplaston of anti-CD146 antibody A A98 and bevacizumab is based on following important scientific discovery: (1) CD146 and VEGFR-2 interact, and are the co-receptors of VEGFR-2; (2) anti-CD146 monoclonal antibody AA98 can block the interaction between CD146 and VEGFR-2, and then blocking VEGF stimulates the activation of VEGFR-2 and the activation of downstream signal (Akt and p38/NF-κ B) causing; (3) migration and the angiogenesis of the endotheliocyte that anti-CD146 monoclonal antibody AA98 inhibition is induced by VEGF; (4), in nude mice lotus tumor model (human pancreatic cancer cell SW1990), AA98 and bevacizumab drug combination more effectively suppress tumor-blood-vessel growth and then suppress tumor growth.
In sum, CD146 can regulate as the co-receptor of VEGFR-2 and the transmission of auxiliary VEGF-VEGFR-2 signal, and then affects migration and the angiogenesis of endotheliocyte.With respect to AA98 or the independent medication of bevacizumab, the anti-CD146 functional antibodies AA98 of our independent development and bevacizumab drug combination can suppress more significantly tumor-blood-vessel growth and then suppress tumor growth in nude mice lotus tumor model.Therefore, we propose targeting can suppress tumor growth more effectively as the co-receptor CD146 of VEGFR-2, and the monoclonal antibody AA98 of anti-CD146 and the pharmaceutical composition of bevacizumab are the novel targeted medicines for the treatment of tumor.AA98 can be by suppressing the interaction between VEGFR-2 and CD146, the activation of the signal path that inhibition VEGF causes, and then blocking-up angiogenesis, and the nutrient supply that cuts off tumor, thus suppress tumor growth.AA98 combines the activation that can more effectively cut off the signal path that determines tumor growth with bevacizumab, suppress more significantly the growth of tumor.
Particularly, the invention provides the following:
1. a compositions that comprises the function fragment of anti-CD146 antibody or described anti-CD146 antibody and the function fragment of VEGF antibody or described VEGF antibody, described compositions is used for suppressing neonate tumour blood vessel, thereby suppresses tumor growth.
2. according to the compositions described in the 1st, wherein said tumor is pancreas tumor.
3. according to the compositions described in the 1st or the 2nd, wherein said anti-CD146 antibody is anti-CD146 monoclonal antibody AA98, and described VEGF antibody is bevacizumab.
According to the compositions described in the 1st in the purposes for the preparation of suppressing in the medicine of tumor growth.
5. according to the purposes described in the 4th, wherein said tumor is pancreas tumor.
6. according to the purposes described in the 4th or the 5th, wherein said anti-CD146 antibody is anti-CD146 monoclonal antibody AA98, and described VEGF antibody is bevacizumab.
Below in conjunction with specific embodiment, the invention will be further described.
Accompanying drawing explanation
In detailed description below in conjunction with accompanying drawing, above-mentioned feature and advantage of the present invention will be more obvious, wherein:
Fig. 1 .CD146 and VEGFR-2 interact at molecular level.A, in Human umbilical vein endothelial cells (HUVECs), co-immunoprecipitation method finds that CD146 and VEGFR-2 interact; B, in HEK293T, the interaction between co-immunoprecipitation method validation CD146 and VEGFR-2; C, external pull-down experiment finds that extracellular region and the VEGFR-2 extracellular region of CD146 exist direct interaction;
Fig. 2. anti-CD146 monoclonal antibody AA98 is by blocking the interaction (A) between CD146 and VEGFR-2, the activation (B-F) of the VEGFR-2 signal path that blocking VEGF is induced;
Fig. 3. level in vitro, AA98 suppresses endothelial cell migration (A) and the angiogenesis (B) of VEGF induction;
Fig. 4. in bearing mouse model, with AA98 or individually dosed group of comparison of bevacizumab, AA98 and bevacizumab drug combination can suppress tumor growth more effectively.(A) different dosing group tumor size schematic diagram; (B) be compared to AA98 or bevacizumab is individually dosed, AA98 and bevacizumab administering drug combinations can suppress the size of tumor more effectively; (C) be compared to AA98 or bevacizumab is individually dosed, AA98 and bevacizumab administering drug combinations can suppress the weight of tumor more effectively; (D) be compared to AA98 or bevacizumab is individually dosed, AA98 and bevacizumab administering drug combinations can suppress tumor-blood-vessel growth more effectively.
The specific embodiment
The antibody A A98 using in this description (Yan, the A novel anti-CD146monoclonal antibody such as X, AA98, inhibits angiogenesis and tumor growth.Blood.2003; 102 (1): 184-191.), AA1 (autonomous production of this laboratory, preservation mechanism: Chinese common micro-organisms preservation center; Preserving number: CGMCC No.2310; Preservation date: 2007.12.28) etc. can obtain according to the description of Chinese Patent Application No. 99107586.2 (CN1234405), Chinese Patent Application No. 200810057260.7 (CN101245101) respectively.Bevacizumab (Bevacizumab) is purchased from Roche company.
Embodiment mono-: cellular level, and CD146 and VEGFR-2 interact;
CD146 and VEGFR-2 are all marks of endotheliocyte, and are the molecules playing a crucial role in angiogenesis, but both contacts unclear.In order to study relation between the two, we utilize the methods such as co-immunoprecipitation to confirm at cellular level, and CD146 and VEGFR-2 exist and interact.Anti-CD146 monoclonal antibody AA98 can block the interaction between CD146 and VEGFR-2.
Main material: Human umbilical vein endothelial cells system (HUVECs) (ATCC, CRL-1730), the HEK293T cell line (ATCC, CRL-11268) of stable transfection CD146cDNA (HEK293T/CD146) etc.
Main agents: cell pyrolysis liquid ，PBS，Shu source anti-CD146 monoclonal antibody AA1 (this laboratory autonomous production.Preservation mechanism: Chinese common micro-organisms preservation center; Preserving number: CGMCC No.2310; Preservation date: 2007.12.28), rabbit anti-VEGFR-2 antibody is (purchased from Cell Signaling Technology company, article No. #2479), His-sCD146 albumen is (purchased from Sino Biological Inc., article No. 50794-M08H), Fc-VEGFR-2 albumen is (purchased from Sino Biological Inc., article No. 10012-H02H), Fc albumen is (purchased from Sino Biological Inc., article No. 10702-HNAH), vegf protein (purchased from Upstate Biotechnology company, article No. B500014).
Main method: co-immunoprecipitation and external pull-down experiment, concrete grammar is as follows:
Co-immunoprecipitation:
1) by 1 * 10
7(Lena Claesson-Welsh et al.EMBO is July 6 J.2005 for the Human umbilical vein endothelial cells of/ml or process transient transfection VEGFR-2 plasmid (10 μ g); 24 (13): HEK293T/CD146 2342-2353.) is inoculated in 100mm culture dish, cell is scraped gently to 4 ℃ of centrifugal 5 minutes centrifugal being collected in Ep pipe after cell density reaches 90%.
2) add 600 μ l lysate RIPA Buffer (50mM Tris-HCl, pH 7.4,1%NP-40,0.25%Na-deoxycholate, 150mM NaCl, 1mM EDTA, 1mM Na3VO4,1mM NaF, (protease inhibitor, purchased from Roche company for 1mM PMSF and 1mM proteinase inhibitors cocktails, article No. 04693116001)), cracking on ice 30 minutes, 4 ℃ centrifugal (12,000g) 15 minutes.
3) drawing supernatant is cell pyrolysis liquid, after Bradford method is measured protein concentration, total protein concentration is diluted to 1mg/ml.
4) in lysate, add 20 μ l protein G-Agarose, hatch 1 hour for 4 ℃, remove the protein with 20 μ l protein G-Agarose non-specific bindings.
5) centrifuging and taking supernatant, adds CD146 monoclonal antibody AA1 or the rabbit anti-VEGFR-2 antibody of 2 μ g, hatches 2 hours for 4 ℃.
6) again add 50 μ l protein G-Agarose, hatch 1 hour for 4 ℃.
7) the centrifugal supernatant of abandoning, the agarose beads of precipitation washes 3 times with the PBS containing protease inhibitor, adds sample-loading buffer 100 μ l vortex 2 minutes after each 5 minutes, and 100 ℃ are boiled 10 minutes, centrifuging and taking supernatant.
8) protein sample of handling well and full cell pyrolysis liquid keep sample and one are used from Western blot and detect.
External pull-down experiment:
1) Fc-VEGFR-2 protein 20 0ng or Fc protein 20 0ng are melted in the EP pipe of 500 μ l PBS together with His-CD146 protein 20 0ng, hatch 1 hour for 4 ℃.
2) add 20 μ l protein G beads, hatch 1 hour for 4 ℃.
3) 4 ℃ centrifugal 5 minutes, abandon supernatant, the agarose beads of precipitation washes 3 times with the PBS containing protease inhibitor, after each 5 minutes, adds sample-loading buffer 50 μ l vortex 2 minutes, 100 ℃ are boiled 10 minutes.
4) protein sample of handling well detects for Western blot.
Result shows as Fig. 1, in HUVECs (A), can be in conjunction with VEGFR-2 when catching CD146 with anti-CD146 antibody A A1, and vice versa, illustrates that CD146 and VEGFR-2 exist to interact.In HEK293T/CD146 (B), in the cell of transfection VEGFR-2, the antibody of rabbit anti-VEGFR-2 can be caught VEGFR-2 and CD146 simultaneously, but in only expressing the cell of CD146, in the result of Western blot, can't detect CD146, this result verification the conclusion of A, CD146 and VEGFR-2 exist and interact.In pull-down experiment, as shown in (C), between CD146 and VEGFR-2, there is direct interaction in vitro.In addition, utilize anti-CD146 monoclonal antibody AA98 or AA1 (100 μ g/ml) to hatch respectively HUVECs, 37 ℃ after 1 hour, with cell culture medium, clean three times, above treated cell is tested for co-immunoprecipitation, as shown in (D), in HUVECs, anti-CD146 monoclonal antibody AA98 can block endogenous CD146 and the interaction between VEGFR-2.These results suggest that CD146 and VEGFR-2 direct interaction in endotheliocyte, anti-CD146 monoclonal antibody AA98 can block interaction between the two.
Embodiment bis-: anti-CD146 antibody is by the interaction between blocking-up CD146 and VEGFR-2, and blocking VEGF stimulates the activation of the VEGFR-2 signal path causing;
Adhesion molecule CD146 has pivotal role in angiogenesis.As the co-receptor of VEGFR-2, CD146 regulates VEGFR-2 signal path.Anti-CD146 monoclonal antibody AA98 can block the interaction between CD146 and VEGFR-2, and then blocking VEGF stimulates the activation of the VEGFR-2 signal path causing.
Utilize anti-CD146 monoclonal antibody AA98 or AA1 (100 μ g/ml) to hatch respectively HUVECs, 37 ℃ after 1 hour, with cell culture medium, clean after three times, then after using VEGF (50ng/ml) to stimulate cell lysis for biochemical analysis signal path.As shown in Fig. 2 (A), in HUVECs, AA98 can block endogenous CD146 and the interaction between VEGFR-2.As shown in (B-F), VEGF stimulation (5 minutes) can cause the phosphorylation of VEGFR-2, and downstream signaling molecule, as Akt (15 minutes), and p38 (15 minutes), the activation of NF-κ B (10 hours).AA98 can block by VEGF stimulates VEGFR-2 and the downstream molecules Akt causing, p38, the activation of NF-kB.Illustrate that CD146 can participate in and regulate the activation of VEGF-VEGFR-2 signal path.
Embodiment tri-: anti-CD146 antibody suppression VEGF stimulates migration and the angiogenesis ability of the endotheliocyte causing;
Angiogenesis is mainly tumor nutrient is provided, and provides approach for what tumor cell shifted, therefore for the growth of tumor, plays vital effect.Suppress angiogenesis and just can suppress significantly the growth of tumor.VEGF is the major cytokine of tumor cell secretion, the signal path final decision tumor-blood-vessel growth of its mediation.CD146 participates in and regulates VEGF-VEGFR-2 signal path, and the antibody suppression VEGF of anti-CD146 stimulates migration and the angiogenesis of the endotheliocyte causing.
Main material: Human umbilical vein endothelial cells is ，96 hole Transwell plate (Corning HTS Transwell-96 Cell Migration Products) etc.
Main agents: VEGF is (purchased from Upstate Biotechnology company, article No. B500014), anti-CD146 antibody A A98, AA1, Mus IgG (mIgG, purchased from Sigma-Aldrich company, article No. I5381), Matrigel is not (containing somatomedin, BD Biosciences, article No. 354234).
Main method: cell migration experiment, endotheliocyte becomes blood vessel experiment.Concrete grammar is as follows:
Cell migration experiment:
1) HUVECs cell is resuspended with complete medium, makes single cell suspension (1 * 10
5/ ml).
2) under Transwell, chamber adds RPMI-1640 culture medium (purchased from Gibco, article No. 31800-022) (200 μl/ hole) the ，Shang chamber containing 10% hyclone to add cell suspension (100 μl/ holes, three parallel holes are established in every kind of processing),
3) upwards in the cell of chamber, add antibody A A98 or AA1 (100 μ g/ml) and stimulant VEGF (100ng/ml).Overnight incubation in 37 ℃ of CO2 gas incubator.
4) cell on Jiang Mo upper strata is wiped with cotton swab, takes ，Mo lower floor off be put on microscope slide upward with the film of tweezers Jiang96 hole transwell plate.
5) lower confluent monolayer cells is fixed after 15 minutes with 4% paraformaldehyde room temperature, and by 1% violet staining 15 minutes, microscopy recorded the cell quantity under each visual field.
It is to improve on the basis of the experimental technique set up people such as Nagata that endotheliocyte becomes blood vessel experiment, and concrete operations are as follows:
1) HUVECs cell is resuspended with complete medium, makes single cell suspension (1 * 10
5/ ml).
2) Matrigel (50 μl/ hole) of coated ice bath in 96 orifice plates, 37 ℃ solidify 30 minutes.
3) in each hole, add 100 μ l cell suspension, correspondingly add stimulant VEGF (100ng/ml) and antibody A A98 or AA1 (100 μ g/ml) simultaneously.
4) overnight incubation in 37 ℃ of incubators is observed afterwards under inverted microscope, takes pictures.
Experimental result as shown in Fig. 3 (A), VEGF can increase endotheliocyte transfer ability and, compare with AA1 matched group, anti-CD146 antibody A A98 can obviously reduce the migration force that VEGF stimulates caused endotheliocyte, suppression ratio is about 53%.Same, with VEGF, stimulate HUVECs can promote HUVECs to form capillary structure.Yet with respect to AA1 processed group, AA98 suppresses the angiogenesis ability that VEGF stimulates caused endotheliocyte significantly.Above result confirmation, the angiogenesis that CD146 mediates for VEGFR-2 is necessary.
Embodiment tetra-: in nude mice lotus tumor model, compare with independent AA98 or bevacizumab, the compositions of AA98 and bevacizumab can suppress more significantly human pancreas cancer tumor-blood-vessel growth and then suppress tumor growth.
In the generation evolution of tumor, angiogenesis provides nutrient for tumor, and provides approach for what tumor cell shifted.Target tumor angiogenesis has become one of main policies for the treatment of clinically tumor at present.Zoopery confirmation, AA98 and bevacizumab drug combination can suppress tumor growth more effectively.
Experimental technique: select 60 4 weeks female BalB/C nude mices (purchased from Beijing Vital River Experimental Animals Technology Co., Ltd.) of size, be divided at random 6 groups and give different antibody with Fen other Give and treat, 10 every group.Respectively at back subcutaneous injection 1 * 10
7individual human pancreatic cancer cell SW1990 cell (ATCC, CRL-2172) (being resuspended in 100 μ l PBS).Treat that gross tumor volume reaches 0.06cm
3, start lumbar injection antibody, be respectively mIgG (purchased from Sigma-Aldrich company, article No. I5381; 200 μ g/ are only), and hIgG (purchased from Zhong Shan Golden Bridge, article No. ZDR-5001; 200 μ g/ are only), AA1 (200 μ g/ are only), AA98 (200 μ g/ are only), bevacizumab is (purchased from Roche company; 200 μ g/ are only), AA98+ bevacizumab (AA98 and bevacizumab respectively 200 μ g/ only, only amount to 400 μ g/), 2 times weekly, measures the length and width of tumor, to calculate gross tumor volume.After 4 weeks, de-neck method is put to death all mices, peels off tumor.After tumor is weighed, with 4%PFA, fix 24 hours, paraffin embedding, section, immunohistochemical analysis.Be below the SABC process of paraffin section:
1) take out slice, thin piece, enter 37 ℃ of dewaxings twice of xylene solution, each 30 minutes;
2) enter aquation in dehydrated alcohol * 2-95%-80%-70%-50%-30% and distilled water, each 5 minutes of room temperature;
3) 37 ℃ of lucifuges of 0.3% hydrogen peroxide/methanol solution are processed 30 minutes, eliminate the activity of endogenous peroxydase, and PBS washes three times;
4) 100 ℃ of water-baths of pH6.0 citric acid repair liquid antigen hot repair in 30 minutes is multiple, natural cooling;
5) 37 ℃ of sealings of 5% normal sheep serum (purchased from company of Zhong Shan Golden Bridge, article No. ZLI-9021) are 1 hour;
6) add PBS dilution primary antibodie (the anti-CD31 multi-resistance of rabbit, purchased from Abcam company, article No. ab28364; Dilution in 1: 50), 4 ℃ of overnight incubation;
7) PBS washes three times; Anti-rabbit-the biotin of goat (purchased from company of Zhong Shan Golden Bridge, article No. ZB-2010; Dilution in 1: 1000) at 37 ℃, hatch 1 hour, PBS washes three times;
8) Avidin-HRP (purchased from Hyclone-pierce company, article No. N100; 1: 1000) at 37 ℃, hatch 45 minutes;
The DAB 9) now joining (purchased from company of Zhong Shan Golden Bridge, article No. ZLI-9032; Dilution in 1: 1000) lucifuge colour developing 2-7 minute, haematoxylin is redyed.
10) dehydration step by step: 50-70-80-90-100-100% ethanol-dimethylbenzene, dries resinene mounting.
11) in micro imaging system, make film.
Result demonstration, as shown in Figure 4, in AA98 and bevacizumab compositions experimental group, compositions is more obvious for the inhibition of tumor growth.Compare with matched group (mIgG or hIgG group), compositions is 70% for the inhibition percentage ratio of gross tumor volume, and the suppression ratio of independent AA98 or bevacizumab only has 46% or 48%.Through the vessel density of the tumor of the compositions-treated of AA98 and bevacizumab still less, be respectively the independent medication group of AA98 or bevacizumab vessel density 40% or 30%.These results suggest that, the compositions of AA98 and bevacizumab can more effectively suppress tumor-blood-vessel growth and then suppress tumor growth.
Claims (6)
1. a compositions that comprises the function fragment of anti-CD146 antibody or described anti-CD146 antibody and the function fragment of VEGF antibody or described VEGF antibody, described compositions is used for suppressing neonate tumour blood vessel, thereby suppresses tumor growth.
2. compositions according to claim 1, wherein said tumor is pancreas tumor.
3. compositions according to claim 1 and 2, wherein said anti-CD146 antibody is anti-CD146 monoclonal antibody AA98, and described VEGF antibody is bevacizumab.
4. compositions according to claim 1 is in the purposes for the preparation of suppressing in the medicine of tumor growth.
5. purposes according to claim 4, wherein said tumor is pancreas tumor.
6. according to claim 4 or purposes claimed in claim 5, wherein said anti-CD146 antibody is anti-CD146 monoclonal antibody AA98, and described VEGF antibody is bevacizumab.
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| PCT/CN2012/080629 WO2014000338A1 (en) | 2012-06-28 | 2012-08-27 | Application of composite of anti-cd146 antibody and anti-vegf antibody in preparing medicine for suppressing tumor |
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| CN102300983A (en) * | 2009-01-30 | 2011-12-28 | 地中海大学 | Human soluble CD146, preparation and uses thereof |
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| TIANXIA JIANG ET.AL: "CD146 is a coreceptor for VEGFR-2 in tumor angiogenesis", 《BLOOD》 * |
| XIYUN YAN ET.AL: "A novel anti-CD146 monoclonal antibody,AA98,inhibits angiogenesis and tumor growth", 《BLOOD》 * |
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| WO2014000338A1 (en) | 2014-01-03 |
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