WO2019047921A1 - Peptide court atr001, anticorps monoclonal préparé par peptide court à fonction de régulation polarisée de l'at1r, et son application - Google Patents

Peptide court atr001, anticorps monoclonal préparé par peptide court à fonction de régulation polarisée de l'at1r, et son application Download PDF

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WO2019047921A1
WO2019047921A1 PCT/CN2018/104608 CN2018104608W WO2019047921A1 WO 2019047921 A1 WO2019047921 A1 WO 2019047921A1 CN 2018104608 W CN2018104608 W CN 2018104608W WO 2019047921 A1 WO2019047921 A1 WO 2019047921A1
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at1r
monoclonal antibody
short peptide
hybridoma cell
atr001
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陈霄
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武汉华纪元生物技术开发有限公司
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/72—Receptors; Cell surface antigens; Cell surface determinants for hormones
    • C07K14/723—G protein coupled receptor, e.g. TSHR-thyrotropin-receptor, LH/hCG receptor, FSH receptor
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/12—Antihypertensives
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/72—Receptors; Cell surface antigens; Cell surface determinants for hormones
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2869—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against hormone receptors
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
    • C07K7/04—Linear peptides containing only normal peptide links
    • C07K7/14—Angiotensins: Related peptides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies

Definitions

  • the present invention relates to short peptides and monoclonal antibodies, in particular to a short peptide ATR001 and to the preparation of monoclonal antibodies and applications for the regulation of AT1R by a short peptide.
  • Renin is a proteolytic enzyme produced and secreted by the para-balloon cells of the kidney, which hydrolyzes angiotensinogen (AGT) to produce angiotensin I (AngI).
  • AGT angiotensinogen
  • AngI angiotensin I
  • Angiotensin-converting enzyme (ACE) widely distributed in tissues such as heart, blood vessels, kidneys, and lungs, can degrade AngI to angiotensin II (Ang II).
  • AngI can also be used in cathepsins, elastase, and chymase.
  • angiotensin-converting enzyme 2 ACE2 can also act on AngII to produce active small molecules such as angiotensin 1-7 (Ang1-7).
  • RAS works in two ways: one is the renin secreted by the renal paracellular cells and the AGT released by the liver enters the blood circulation, and a small amount of active small molecule fragments are generated by a series of enzyme hydrolysis to exert physiological effects. The other is to produce an active molecule such as AngII locally in a paracrine manner. It is currently believed that AngII is the most important bioactive peptide in RAS and binds to the corresponding receptor to exert biological effects. The AT1 receptor (AT1R) is the most important AngII receptor.
  • AT1R As a G-protein coupled receptor (GPCR), AT1R has a typical seven-transmembrane alpha helix structure, which is mainly distributed in tissues and organs such as blood vessels, heart, kidney, and fat. The combination of AngII and AT1R plays the following roles:
  • AT1R plays an important role in the regulation of blood pressure in human body and plays an important role in the occurrence and development of hypertension.
  • AngII can also stimulate the release of endothelin, which ultimately leads to the proliferation and contraction of vascular smooth muscle, endothelial damage, and remodeling of blood vessels, which leads to the development and progression of hypertension.
  • AT1R is also involved in the development and progression of atherosclerosis.
  • Wassmann et al also confirmed that apoE and AT1R double knock mice can effectively block vascular oxidative stress, endothelial dysfunction, atherosclerotic lesion formation caused by high cholesterol diet, and independent of blood pressure and blood lipid levels.
  • AngII activates NA2-/NADPH oxidase on the cell membrane of vascular endothelial cells through AT1R, which produces O 2- , which in turn activates NF ⁇ B, initiates MCP-1 expression, and promotes the accumulation of monocytes/macrophages in the blood vessel wall. It migrates to the endothelium, then changes to the inflammatory phenotype, and participates in early lesions of atherosclerosis such as foam cell formation.
  • AT1R abdominal aortic aneurysm
  • apoE -/- or Ldlr -/- pumped high concentrations of AngII showed tube diameter dilatation, thrombosis, vascular smooth muscle apoptosis and loss, elastic fiber rupture, extracellular matrix deposition, a large number of inflammatory cell infiltration of the adventitia, Similar changes in human abdominal aortic aneurysm lesions such as MMPs/TIMPs imbalance, and the above changes were not related to blood pressure and blood lipid levels.
  • ACEI angiotensin-converting enzyme inhibitors
  • ARB angiotensin 1 receptor antagonist
  • GPCRs can be coupled with a variety of downstream G proteins, even non-G proteins (such as ⁇ -arrestin), and ligands can selectively make GPCRs when interacting with GPCRs.
  • Certain G proteins or ⁇ -arrestins are coupled to certain signaling pathways, referred to as ligand bias.
  • ligand bias Taking AT1R as an example, after binding of AngII, PKC can be activated by coupling Gq to increase intracellular calcium, and then the downstream phosphorylase activation pathway is initiated.
  • Gq activation can phosphorylate the intracellular portion of AT1R by GPCR-associated protein kinase 5 (GRK5), and ⁇ -arrestin2 binds to it, and is degraded or recycled to the cell surface by ⁇ -arrestin2-mediated desensitization.
  • GPCR-associated protein kinase 5 GRK5
  • ⁇ -arrestin2 binds to it, and is degraded or recycled to the cell surface by ⁇ -arrestin2-mediated desensitization.
  • ⁇ -arrestin activation can also initiate downstream signal transduction, such as ERK activation and is not affected by the Gi protein inhibitor PTX.
  • the binding of Ang II and AT1R can also be coupled to Gi, Gi as an inhibitory regulation of GPCR, and also considered to be involved in receptor internalization desensitization and intracellular phosphorylase activation. way.
  • the biasing regulation of receptor action is more precise and can play a role that needs to be exerted, while avoiding the occurrence of adverse effects caused by total blockage.
  • the regulation of receptor bias is a hot spot in drug development, but there is no bias to regulate AT1R drug development.
  • the extracellular second loop of AT1R is critical for receptor function, and early studies have shown that autoantibodies against the extracellular second loop of AT1R can be detected in the serum of patients with pre-eclampsia and refractory hypertension, which can aggravate vascular damage. To promote the development of hypertension.
  • drugs that can block the activation of AT1R and bias the regulation of AT1R we screened the extracellular epitope of AT1R and screened the antibodies against the selected AT1R target sequences. The effects were confirmed in animal models of hypertension, atherosclerosis and aneurysms.
  • the present invention provides a short peptide ATR001 and a monoclonal antibody and application for preparing a biased regulatory AT1R function from a short peptide.
  • the invention adopts the monoclonal antibody technology to prepare a monoclonal antibody against AT1R, and the antibody can bias the AT1R function, and exerts a therapeutic effect in vascular remodeling diseases such as hypertension, atherosclerosis and aneurysm.
  • the present invention protects a short peptide ATR001 which produces hybridoma cell CQ8-A2D9, the amino acid sequence of which is AFHYESQ.
  • the present invention also protects a hybridoma cell CQ8-A2D9, which has the accession number: CCTCC NO: C2017108.
  • the above hybridoma cell CQ8-A2D9 was deposited with the China Center for Type Culture Collection (CCTCC) on November 16, 2016; the deposit number is CCTCC NO: C2017108; the deposit location is: China, Wuhan, Wuhan University.
  • CTCC China Center for Type Culture Collection
  • the invention also protects a preparation method of the above hybridoma cell CQ8-A2D9, comprising the following steps:
  • the short peptide ATR001 is coupled to the carrier protein KLH as an immuno-coupled antigen
  • the immunoconjugated antigen is mixed with Freund's adjuvant to immunize the mouse, and then the spleen cells of the immunized mouse are fused with the myeloma cell SP2/0, and the hybridoma cell CQ8-A2D9 is obtained by screening.
  • the amino acid sequence of the short peptide ATR001 is AFHYESQ
  • the present invention also protects the above hybridoma cell CQ8-A2D9 for the preparation of the monoclonal antibody anti-AT1R, the hybridoma cell CQ8-A2D9 has the accession number: CCTCC NO: C2017108.
  • the principle of monoclonal antibody antibody technology is that B lymphocytes can produce antibodies, but incapable of infinite division in vitro; while tumor cells can be infinitely passaged in vitro, but can not produce antibodies.
  • the hybridoma cells obtained by fusing these two cells have the characteristics of two parental cells. It is capable of generating highly specific and high affinity antibodies against a single antigenic epitope.
  • the above method prepares a monoclonal antibody anti-AT1R having a bias-regulating AT1R function, and the amino acid sequence of the monoclonal antibody anti-AT1R includes a heavy chain or a light chain, the sequences of which are SEQ ID No. 1 and SEQ ID No. 2, respectively. .
  • the present invention also contemplates the use of the monoclonal antibody anti-AT1R prepared by the above-described preparation to have a bias-regulating AT1R function for the treatment of hypertension, atherosclerosis and aneurysms.
  • the beneficial effects of the present invention are that the monoclonal antibody anti-AT1R prepared by ATR001 can preferentially regulate AT1R, thereby exerting a protective effect in vascular remodeling diseases (hypertension, atherosclerosis, aneurysm).
  • vascular remodeling diseases hypertension, atherosclerosis, aneurysm
  • Figure 1 shows the effect of anti-AT1R antibody on AngII-induced ERK phosphorylation
  • Figure 2 shows the effect of anti-AT1R antibody on AngII-induced JNK phosphorylation
  • FIG. 3 shows the effect of anti-AT1R antibody on the change of intracellular calcium concentration induced by AngII.
  • AT1R-antibody is anti-AT1R;
  • Figure 4 is a graph showing the serum antibody titer of mice in the monoclonal antibody group on the second day of monoclonal antibody injection;
  • Figure 5 is a graph showing changes in blood pressure of mice during the experiment.
  • FIG. 6 is a comparison of the monoclonal antibody anti-AT1R aorta gross atherosclerotic plaque.
  • mAb refers to anti-AT1R antibody
  • Val refers to valsartan
  • Con is a control group
  • FIG. 6A is a general view of vascular lesions
  • FIG. 6B is a statistical diagram of the area of vascular gross lesions
  • Figure 7 shows the area of the aortic valve plaque of the monoclonal antibody anti-AT1R compared with the control group.
  • mAb refers to anti-AT1R antibody
  • Val refers to valsartan
  • Con is a control group
  • Figure 7A is a sectional view of aortic annulus
  • Figure 7B is a statistical diagram of aortic annulus lesions
  • Figure 8 is a comparison of monoclonal antibody anti-AT1R on AAA tumor formation rate and tumor size, C is control, S is AngII perfusion group, A is AngII perfusion plus anti-AT1R antibody;
  • Fig. 8A is a general view of an aneurysm
  • Fig. 8B is a statistical diagram of the incidence of an aneurysm
  • Figure 8C is a statistical diagram of the diameter of the aneurysm
  • Figure 8D is a statistical classification of the aneurysm pathology
  • Figure 9 is a schematic diagram showing the integrity of the monoclonal antibody anti-AT1R to protect the vascular wall, C is the control, S is the AngII perfusion group, and A is the AngII perfusion plus anti-AT1R antibody group.
  • the experimental methods in the following examples are conventional methods unless otherwise specified.
  • the test materials used in the following examples, unless otherwise specified, were purchased from a conventional biochemical reagent store.
  • the quantitative tests in the following examples were set up with three replicate experiments, and the results were averaged, unless otherwise specified.
  • the conventional pharmaceutical preparations in the examples are as follows:
  • the cell culture medium used in the following experiments mainly consisted of two basic media, RPMI-1640 or DMEM. After being prepared, the cells were sterilized by filtration (0.22 um), and stored at 4 ° C.
  • the incomplete RPMI-1640 medium formulation is:
  • Complete RPMI-1640 or DMEM medium incomplete RPMI-1640 or DMEM medium 80ml + calf serum 15-20ml;
  • HT medium formula complete RPMI-1640 or DMEM medium 99ml + HT stock solution 1ml;
  • HAT medium formula complete RPMI-1640 or DMEM medium 98ml + HT stock solution 1ml + A stock solution 1ml
  • aminoguanidine (Aminopterin MW 440.4) was weighed and dissolved in 90 ml of ultrapure water or four distilled water, neutralized by adding 1 mol/L NaOH 0.5 ml, and then added with ultrapure water or four distilled water to 100 ml. Filter and sterilize, dispense vials (2ml/bottle), and store at -20 °C.
  • F.7.5% NaHCO 3 solution Weigh analytically pure NaHCO 3 7.5g, dissolve in 100ml ultrapure water or four distilled water, filter and sterilize, dispense vial (4-5ml/bottle), close the stopper, 4°C save.
  • HEPES N-2-Hydroxyethylpiperazine-N,-2-ethanesulfonic acid, N-2-hydroxyethylpiperazine-N,-2-ethylsulfonic acid, MW 238.3 in 100ml of ultrapure water or In four distilled water, filter and sterilize, dispense vials (4-5ml/bottle), and store at 4°C.
  • the short peptide ATR001 with a purity of ⁇ 85% of the peptide synthesis is carried out for the sequence AFHYESQ, and then the short peptide ATR001 is coupled with the carrier protein KLH as an immunizing antigen;
  • mice 6-week-old Balb/c mice (live mice) were immunized with immunizing antigens, regular cycle: 3 weeks - 2 weeks - 2 weeks... Intensive cycle: 2 weeks - 2 weeks - 1 week...
  • Mouse antiserum ELISA titer ⁇ 1:80000 the specific steps are as follows:
  • the dose is 50ug, intraperitoneal injection
  • mice spleen fusion was performed, and mouse spleen cells with titer ⁇ 1:80000 were taken.
  • the preparation method of myeloma cell suspension is as follows:
  • Bone marrow cells are expanded and cultured 48-36 hours before the fusion (generally, about 2-3 bottles of 100 ml culture flasks are prepared by a 96-well fusion test);
  • the cells are gently blown from the bottle wall tumor with a curved pipette and collected in a 50 ml centrifuge tube or a fusion tube;
  • mice that had been immunized were taken, the eyeballs were removed, and the serum was separated as a positive control serum at the time of antibody detection.
  • the mice were sacrificed by cervical dislocation, soaked in 75% alcohol for 5 minutes, fixed on the culture dish, and then the left abdomen skin was opened. The spleen was visible, and the eye scissors were cut. The scissors were opened in a clean bench. The peritoneum was taken out and placed in a dish containing 10 ml of incomplete medium, gently washed, and the surrounding connective tissue was carefully peeled off.
  • the plate was placed in a stainless steel sieve, and the cell suspension was ground with a syringe needle and counted to allow the spleen cells to enter the incomplete medium in the plate. Sip several times with a pipette to make a single cell suspension. Usually 1 x 10 8 - 2.5 x 10 8 splenocytes per mouse.
  • mice After the mice were sacrificed, the surface was disinfected and fixed, the abdominal skin was lifted from the posterior abdomen with a sterile scissors to expose the peritoneum. Wipe the peritoneum with an alcohol cotton ball. Inject 10 ml of incomplete medium into the abdominal cavity with a syringe, taking care to avoid penetrating the intestine. Fix the syringe in the right hand, leave the needle in the abdominal cavity, gently massage the abdomen with an alcohol cotton ball for 1 minute, then aspirate the injected culture solution. Centrifuge at 1000 r/min for 5-10 minutes and discard the supernatant.
  • the precipitated cells were first suspended in 5 ml of HAT medium, and HAT medium was added according to the cell count to make the cell concentration 2 ⁇ 10 5 /ml, and was used.
  • HAT medium for macrophages, a 96-well plate requires 2 x 10 4 cells per well, and a 24-well plate requires 105 cells per well.
  • Each mouse can obtain 3-5 ⁇ 10 6 cells, so one mouse can be used for feeding cells of two 96-well plates.
  • the feeder cells can also be prepared and cultured 1-2 days before cell fusion, such that the bottom of the culture plate is first covered with a layer of feeder cells.
  • the above cell suspension was added to a 96-well plate at 0.1 ml per well, and then cultured in an incubator at 37 ° C in a 6% CO 2 atmosphere.
  • the selection medium was added, 50ul per well, and after 3 days, the medium was replaced with a half medium;
  • 1 antigen is diluted to 10ug/ml with a coating solution
  • each hybridoma cell is divided into 96-well plates, each well is 100ul;
  • the cells were cultured at 537 ° C, 5% CO 2 for 6 days, and the clones were detected by macroscopic detection; under the inverted microscope, only the wells of single clone growth were marked, and the supernatant was taken for antibody detection.
  • Hybridoma cells diluted with PBS or serum-free medium were inoculated intraperitoneally after 27-10 days, 5 ⁇ 10 5 /0.2 ml per mouse.
  • mice After 5 days, observe the ascites production in mice every day. If the abdomen is obviously enlarged, when the skin is touched by hand, the skin can be used to collect ascites with a 16-gauge needle. Generally, it can be taken continuously for 2-3 times, usually for each mouse. Can take 5-10ml ascites;
  • the ascites was centrifuged (2000 r/min for 5 minutes) to remove the cellular components and other precipitates, and the supernatant was collected, and the antibody titer was determined, and the mixture was stored at -70 ° C for storage or stored by lyophilization.
  • the monoclonal antibody anti-AT1R is sequenced, and the amino acid sequence of the monoclonal antibody anti-AT1R includes a heavy chain or a light chain, and the sequences thereof are SEQ ID No. 1 and SEQ ID No. 2, respectively.
  • AT1R-CHOK1 cell line (PerkinElmer): DMEM/F12 complete medium containing 10% FBS, cultured in a 5% CO 2 cell incubator;
  • AT1R-CHOK1 was inoculated into a six-well plate at a density of about 80% and cultured overnight. Then, HBS was starved for 2 h, AngII concentration gradient stimulation and time gradient stimulation were performed, and the expression of p-ERK/ERK and p-JNK/JNK was detected by Western Blot.
  • Fig. 1 AngII-induced ERK
  • Fig. 2 JNK phosphorylation
  • Fig. 3 the monoclonal antibody anti-AT1R had no effect on the increase of intracellular calcium concentration induced by AngII (Fig. 3), namely the monoclonal antibody anti-AT1R.
  • a blood pressure model was constructed by instilling AngII into the Balb/c mouse subcutaneously.
  • the monoclonal antibody group (group A) was injected with anti-AT1R monoclonal antibody 100 ⁇ g/only in the tail vein of the buried pump, and the hydralazine group (group J) was used.
  • gavage was administered at 3 mg/kg/d.
  • a Western diet-induced atherosclerosis (AS) mouse model was established, in which apoE -/- mice were fed with high fat for 17 weeks, and anti-AT1R monoclonal antibody (mAb) was 20ug and 100ug per week.
  • mAb monoclonal antibody
  • the apoE -/- mouse aorta was roughly stained with oil red after 17 weeks, and the area of atherosclerotic plaque was significantly decreased (Fig. 6).
  • the aortic annulus was stained with oil-red stained in frozen sections, and the anti-AT1R monoclonal antibody was found to significantly reduce atherosclerotic plaques (Fig. 7).
  • an AngII-induced abdominal aortic aneurysm (AAA) mouse model was constructed, in which apoE -/- mice were subcutaneously implanted with a micro-osmotic pump and AngII was administered at 1000 ng/kg/min for 28 days.
  • the monoclonal antibody anti-AT1R was found to be significant. Reduce the incidence of AAA in apoE -/- mice and reduce the severity of the tumor ( Figure 8). Cross-sectional pathology of the tumor vessels revealed a significant expansion of the control vessels with thrombosis, while the monoclonal antibody anti-AT1R protected the vascular morphology and morphology (Fig. 9).

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Abstract

La présente invention concerne un peptide court ATR001, un anticorps monoclonal préparé par un peptide court et ayant une fonction de régulation polarisée de l'AT1R, ainsi qu'une application de celui-ci ; le peptide court ATR001 présente une séquence d'acides aminés de AFHYESQ et une cellule d'hybridome CQ8-A2D9, le numéro de dépôt de celui-ci est : CCTCC NO : C20 17108. La présente invention utilise la méthode des hybridomes pour la préparation d'un anticorps monoclonal ciblant AT1R, et l'anticorps peut avoir une fonction de régulation polarisée de l'AT1R, ayant un effet thérapeutique dans des maladies de remodelage vasculaire telles que l'hypertension, l'athérosclérose et les anévrismes.
PCT/CN2018/104608 2017-09-08 2018-09-07 Peptide court atr001, anticorps monoclonal préparé par peptide court à fonction de régulation polarisée de l'at1r, et son application WO2019047921A1 (fr)

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CN201710807113.6A CN109467599B (zh) 2017-09-08 2017-09-08 短肽atr001以及由短肽制备具有偏向性调节at1r功能的单克隆抗体和应用
CN201710807113.6 2017-09-08

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