CN114569745A - PD-L1-targeted polypeptide PET molecular imaging probe and preparation method and application thereof - Google Patents
PD-L1-targeted polypeptide PET molecular imaging probe and preparation method and application thereof Download PDFInfo
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- C—CHEMISTRY; METALLURGY
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- A61K51/0474—Organic compounds complexes or complex-forming compounds, i.e. wherein a radioactive metal (e.g. 111In3+) is complexed or chelated by, e.g. a N2S2, N3S, NS3, N4 chelating group
- A61K51/0482—Organic compounds complexes or complex-forming compounds, i.e. wherein a radioactive metal (e.g. 111In3+) is complexed or chelated by, e.g. a N2S2, N3S, NS3, N4 chelating group chelates from cyclic ligands, e.g. DOTA
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- A—HUMAN NECESSITIES
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- A61K51/00—Preparations containing radioactive substances for use in therapy or testing in vivo
- A61K51/02—Preparations containing radioactive substances for use in therapy or testing in vivo characterised by the carrier, i.e. characterised by the agent or material covalently linked or complexing the radioactive nucleus
- A61K51/04—Organic compounds
- A61K51/08—Peptides, e.g. proteins, carriers being peptides, polyamino acids, proteins
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Abstract
The invention relates to the technical field of radiopharmaceutical chemistry and clinical nuclear medicine, in particular to a polypeptide PET molecular imaging probe targeting PD-L1, and a preparation method and application thereof.18The F-AlF-NOTA-PA2881 probe is a cyclic polypeptide PET molecular imaging probe consisting of ten amino acids based on Cyclopeptide 66, and is characterized in that a bifunctional chelating agent is connected to lysine in cyclic peptide and then radionuclide is used for carrying out secondary hybridization on the cyclic peptide18And F is marked. Using short half-life radionuclides18F marking can quickly and accurately identify the tumor focus. The molecular imaging probe precursor is a ringCompared with the existing mainstream monoclonal antibody imaging probe, the peptide cyclopoptide 66 is rapidly metabolized, and the potential influence of the radionuclide and the monoclonal antibody on organisms is further reduced. The in vitro imaging device PET can reflect the expression level of the tumor cell PD-L1, and the immunotherapy curative effect, real-time curative effect detection, estimation prognosis and the like aiming at the signal channel can be predicted. The probe has excellent in-vivo targeting performance.
Description
Technical Field
The invention relates to the technical field of radiopharmaceutical chemistry and clinical nuclear medicine, in particular to a polypeptide PET molecular imaging probe targeting PD-L1, and a preparation method and application thereof.
Background
In the prior art, immune checkpoints are signaling pathways that play an important role in mediating immune tolerance and preventing excessive immune responses, where programmed death receptor-1 (PD-1)/programmed death ligand 1 and 2(PD-L1/2) signaling pathways can be expressed in a variety of malignant tumor tissues, affecting antigen signaling presentation. PD-1 is a negative co-stimulatory receptor expressed on the surface of activated T cells and primary B cells, PD-1 has two ligands (PD-L1/2), PD-L1 is a type 1 transmembrane surface glycoprotein ligand widely expressed in T cells, B cells and macrophages, PD-1 interacts with PD-L1 to negatively regulate immunity through the regulation of T lymphocyte proliferation, cytokine production and cytotoxic activity to place CD28/MHC in an activated state and maintain the homeostasis of the immune system. In tumor tissues, PD-L1 is also overexpressed on tumor cells and mediates apoptosis of antigen-specific effector T cells through interaction with PD-1, and the targeted combination of the two promotes immunosuppression of antigen presenting cells and tumor cells.
The tumor cells create an immunosuppressive environment through the activation of a PD-1/PD-L1 signal channel, thereby avoiding the immune surveillance of a host, realizing the immune escape and inducing the unlimited proliferation of the tumor cells. Therefore, the blocking of the immune checkpoint can be realized by using the immune checkpoint inhibitor, the negative immune regulator on the tumor cell or the receptor thereof on the immune effector cell is inhibited, and the immune system is activated to kill the tumor cell in a targeted way. This immunotherapy against the PD-1/PD-L1 signaling pathway is becoming an increasingly common treatment for various cancers, and PD-1/PD-L1 immune checkpoint inhibitors not only improve overall survival, but maintain a sustained response and reduce the incidence of adverse events compared to traditional chemotherapy. First generation immune checkpoint inhibitors are immunomodulatory monoclonal antibodies that block immune checkpoints, since the FDA approved the first antibodies targeting checkpoint receptor CTLA-4 for use in combination with anti-cancer therapy in 2011, the advent of PD-1-targeted monoclonal antibodies (nivolumab and pembrolizumab) and PD-L1-targeted monoclonal antibodies (avelumab, atezolizumab and durvalumab) in tandem have been approved for clinical immunotherapy of a variety of malignancies.
For the pre-immunotherapy prediction of therapeutic effect, immunohistochemistry, Polymerase Chain Reaction (PCR) -based methods, and detection of serum or blood biomarkers are generally used clinically. Although these methods have been widely used, it remains a great challenge to correctly select patients suitable for immunotherapy and to accurately assess the response to therapy. In addition, there are increasing reports of immune related adverse events. Currently, there is no reliable monitoring strategy to diagnose these unexpected complications. There is increasing evidence that dynamic visualization of systemic immune responses by PET (positron emission tomography) can effectively guide and monitor clinical immunotherapy. To achieve this goal, radioactive molecular imaging probes were developed for PET imaging.
Recently, the development speed of PET immune targeting molecular imaging probes based on monoclonal antibodies is high, and several molecular imaging probes targeting PD-L1 have been reported in the latest literature to achieve clinical transformation. These drugs provide the affinity and specificity of the maximum signal-to-background ratio required to visualize molecular targets in the tumor microenvironment. However, a major disadvantage of antibody-based molecular imaging probes is their slow rate of clearance. Typically, clearance of antibodies through the hepatobiliary system takes several days to weeks before they are metabolized out of the body, making imaging of the adjacent organs of the liver, such as the lung, pancreas, etc., a challenge. Therefore, it takes many days for the molecular imaging probe to be sufficiently cleared from the non-target tissue or the tissue adjacent to the tumor to achieve imaging. The target PD-L1PET molecular imaging probe has higher tumor uptake rate in PD-L1 positive tumors and low signal background in non-PD-L1 expression tissues, and can enable patients to flexibly image the day in clinical research design, so that the target PD-L1PET molecular imaging probe is ideal.
Cyclopeptide 66 is a 10 amino acid cyclic polypeptide (-leucine-D-tryptophan-proline-threonine-aspartic acid-leucine-D-phenylalanine-lysine-valine-arginine-), and hydrophilic and hydrophobic residues on both sides of the molecular ring show stable beta-sheet conformation, forming a unique amphiphilic molecule. The amino acid sequence of the Cyclopeptide 66 is as follows: cyclo (-Leu-DTrp-Pro-Thr-Asp-Leu-DPhe-Lys-Val-Arg-), Cyclopeptide 66 has been disclosed in Sun H, Chen D, Zhan S, Wu W, Xu H, Luo C, Sun H, Feng Y, Shao W, Wan A, ZHou B, Wan G, Bu X. Design and Discovery of Natural Cyclopeptide Skeleton Based Programmed depletion Ligand 1 Inhibitor as Immortor Modulator for Cancer therapy J Med chem. 2020 Oct 8;63(19): 11286. supplement 11301. can be obtained by chemical synthesis. The molecule can show good capacity of combining PD-L1, can effectively block a PD-1/PD-L1 signal channel, induces immune cells to a tumor microenvironment, enhances the killing of the immune cells on the tumor cells by promoting the release of granzyme B and perforin, and shows remarkable CD8 in vivo+T cell dependent tumor suppressor activity.
The structural formula of the Cyclopeptide 66 is as follows:
in view of the fact that the Cyclopeptide (Cyclopeptide 66) targeting PD-L1 has small molecular metabolic advantages and low half inhibitory concentration (IC50=1.42 mu M), the invention constructs a positron nuclide based on the Cyclopeptide 6618F-labeled PD-L1 targeted molecular probe18F-AlF-NOTA-PA2881, and the preclinical verification of PD-L1 positive/negative expression tumor model in vivo identification by PET animal in vivo imaging (higher signal-to-noise ratio can be obtained, and tumor regions can be accurately identified), has great significance for estimation of curative effect of PD-L1 immunotherapy of malignant tumor patients, monitoring and evaluation of curative effect of immunotherapy drugs, prognosis evaluation and the like, and has great clinical transformation prospect.
Disclosure of Invention
The invention aims to provide a polypeptide PET molecular imaging probe with excellent in-vivo targeting performance and targeting PD-L1, and a preparation method and application thereof.
The technical scheme of the invention is as follows:
polypeptide PET molecular imaging probe targeting PD-L1 (PD-L1 targeting PET molecular imaging probe)18F-AlF-NOTA-PA 2881) characterized by the structural formula:
a method for preparing the polypeptide PET molecular imaging probe targeting PD-L1 according to claim 1, characterized by the following steps:
1) reacting Cyclopeptide 66 with a bifunctional chelating agent to obtain a product (the feed ratio is 1-1.5); the bifunctional chelating agents include, but are not limited to, NOTA, DOTA or NODAGA.
2) Labeling the product obtained in the step 1) with radionuclide.
The radionuclides include, but are not limited to18F、68Ga、64Cu or89Zr。
The method for labeling the radionuclide comprises the following steps: dissolving the product of the step 1) in DMSO or deionized water, 10 mM, 2-4 μ L; at the placeAdding AlCl into the obtained solution30.1M sodium acetate aqueous solution (2 mM, 5-10 μ L), 200-.
The radionuclides include, but are not limited to18F、68Ga、64Cu or89Zr. Preferably, it is18F、68Ga or64And (3) Cu. Further preferred radionuclides are18F。
The bifunctional chelating agents include, but are not limited to, NOTA, DOTA or NODAGA; further the bifunctional chelating agent is NOTA.
The method for labeling the radionuclide comprises the following steps: dissolving the product of the Cyclopeptide 66 and NOTA connection in DMSO or acetonitrile at 10 mM and 200-; adding AlCl32mM, 5-10 μ L of sodium acetate; adding Na purified by QMA column (anion exchange column)18Solution F, 200-; heating to 100 deg.C under sealed condition, reacting for 10-20min, cooling, passing the reaction solution through pre-activated C18-light solid phase extraction column, washing with deionized water to remove unreacted free substance18F ion, eluting the product from C18-light solid phase extraction column with 75% ethanol solution to obtain the compound represented by the formula18F, diluting the ethanol concentration to be below 10% to prepare the tracer injection.
Tracer agent18The F-AlF-NOTA-PA2881 is used as a PD-L1 molecular targeted tracer in PET molecular imaging.
Particularly as a tracer for detecting the expression condition of PD-L1 of tumors in patients with solid tumors (non-small cell lung cancer, gastric cancer and colorectal cancer), the tracer can be specifically combined with PD-L1 protein on the tumor cell membranes and can be qualitatively and quantitatively analyzed by various positron nuclide detection devices.
There is provided the use of a radiolabelled cyclic peptide compound, in particular a tracer, prepared by the above method18PET imaging application of F-AlF-NOTA-PA 2881.
The abbreviations used in the present invention represent the following meanings, respectively:
DIPEA: n, N-diisopropylethylamine.
HEPES-EDTA: 4- (2-hydroxyethyl) -1-piperazineethanesulfonic acid-ethylenediaminetetraacetic acid.
DMSO, DMSO: dimethyl sulfoxide (DMSO).
18The F-AlF-NOTA-PA2881 probe is a cyclic polypeptide PET molecular imaging probe consisting of ten amino acids based on Cyclopeptide 66, and is formed by connecting a bifunctional chelating agent (such as NOTA) to lysine in cyclic peptide and then passing through radionuclide18And F is marked.
Or the name is:18f-labeled PD-L1-targeted PET molecular imaging probe18F-AlF-NOTA-PA2881, and a preparation method and application thereof.
The invention has the beneficial effects that: 1. using short half-life radionuclides18F (T1/2-109 min) mark can quickly and accurately identify the tumor focus (0.5 hour). 2. Compared with the existing mainstream monoclonal antibody imaging probe, the molecular imaging probe precursor which is Cyclopeptide 66 is quickly metabolized, and the potential influence of radioactive nuclides and monoclonal antibodies on organisms is further reduced. The in vitro imaging device PET can reflect the expression level of the tumor cell PD-L1, and the immunotherapy curative effect, real-time curative effect detection, estimation prognosis and the like aiming at the signal channel can be predicted. 3. The in vivo targeting performance of this probe was excellent (see example 2).
Embodiments of the present invention will be described in further detail below with reference to the accompanying drawings and examples.
Drawings
FIG. 1 shows the tumor-bearing nude mouse model injected with different PD-L1 expression level cell lines (HCC 827, H460) with the present invention18Representative coronal PET images after F-AlF-NOTA-PA2881, with arrows indicating tumor regions.
Fig. 2 is a graph of the result of quantitative analysis of the image of fig. 1.
Detailed Description
Example 1
PD-L1-targeted polypeptide PET molecular imaging probe (A)18F-labeled PD-L1-targeted PET molecular imaging probe18F-AlF-NOTA-PA 2881) comprises the following steps:
1) preparation of NOTA-PA 2881: 1. dissolving 5mg of Cyclopeptide 66 in HEPES-EDTA solution, adding 3 equivalents of triisopropylacetamide (2-carboxyethyl) phosphine (dissolved in 0.2 mol/L ammonium acetate buffer, pH = 7) thereto, evacuating the solution of gas, and repeating three times; reacting at room temperature for 60 minutes, then transferring to a centrifugal filter, and centrifuging for 90 minutes at 4000 revolutions per minute; washing the solution with 1mL of 0.2 mol/L ammonium acetate buffer, adding the reduced polypeptide into 2 mL of 0.2 mol/L ammonium acetate buffer (pH = 7), and transferring to a second reaction vessel; 4 mg of NOTA-maleimide and 0.5 mL of 0.2 mol/L sodium acetate buffer (pH = 7) were mixed and added to the second reaction vessel, followed by reaction at 40 ℃ for 3 hours; after the reaction was completed, the reaction mixture was transferred to a centrifugal filter and centrifuged at 4000 rpm for 90 minutes; after discarding the flow, adding 2 mL of ultrapure water, centrifuging for 90 minutes again at 4000 rpm, and discarding the flow again; collecting purified NOTA-PA2881 by using 1mL of ultrapure water, freeze-drying, and confirming by mass spectrometry to obtain a NOTA-PA2881 compound which is a precursor PA2881 structural formula, namely a structural formula of a NOTA connecting product of Cyclopeptide 66 and a bifunctional chelating agent:
2) preparation of18F-AlF-NOTA-PA2881。
To a 1.5mL EP tube was added 300. mu.L acetonitrile, 5. mu.L of 2mM AlCl3Adding 2 mu L of 10 mM DMSO or deionized water dissolved cyclopoptide 66 into 0.1M sodium acetate aqueous solution, finally adding the QMA column purified target water which is well activated, and leaching with physiological saline to obtain Na of 1.85-3.70 GBq (50-100 mCi)18Solution F400. mu.L. The mixture is shaken up and reacts for 10-15min at 100 ℃ under the closed condition, and is cooled to room temperature.
The reaction solution was slowly injected into a previously activated Sep-Pak C18-light column, and washed with 10mL of deionized water to remove free18F ions and water-soluble impurities are dried and then leached by 500 mu L of 75 percent ethanol water solution to obtain the compound shown in the structural formulaIs/are as follows18Diluting eluate with normal saline to ethanol content less than 10%, measuring radiochemical purity by radioactive HPLC, observing appearance as colorless clear transparent liquid, decay correcting, and probing18The synthetic yield of F-AlF-NOTA-PA2881 is more than or equal to 25 percent, and the radiochemical purity is more than or equal to 95 percent.
18F-AlF-NOTA-PA2881 structural formula:
example 2
Probe needle18In vivo targeting validation of F-AlF-NOTA-PA 2881.
In-vivo targeting verification experiment, cell lines HCC827 and H460 with high PD-L1 expression levels and low PD-L1 expression level are selected to establish a nude mouse tumor model (female mouse, 5 weeks old, about 20g, 2X 106The cells are planted in the right shoulder under the skin, and the tumor volume is about 300mm after about 4 weeks3In vivo imaging experiments) by rat tail injection18Following the F-AlF-NOTA-PA2881 probe solution (injection dose was 9.25MBq, 150. mu.L), a PET/CT scan (Discovery 790Elite, GE Healthcare) was performed for 0.5 hour, 1 hour, 5min scan duration under isoflurane-oxygen anesthesia, image post-processing platform AW4.6 software (GE Healthcare).
Imaging results as shown in fig. 1, HCC827 tumor uptake probe was significantly higher than background and significantly higher than H460, indicating that the probe18F-AlF-NOTA-PA2881 can specifically identify a tumor model with high expression of PD-L1, and a tumor area with higher resolution can be observed from a whole-body image.
Results of image quantitative analysis FIG. 2 shows that at 0.5 and 1 hour time points, the uptake of probe by HCC827 tumors was 4.75. + -. 0.31% ID/g and 3.71. + -. 0.33% ID/g, respectively, whereas the uptake of probe by H460 tumors was 1.96. + -. 0.56% ID/g and 1.58. + -. 0.61% ID/g, both of which were significantly lower than the former, and the differences were statistically significant (P.sup.5 and 1 hour time points)<0.05), further indicating that the probe can be specifically taken up18F-AlF-NOTA-PA2881, namely the probe can specifically accumulate in HCC827 tumor area with high expression of PD-L1, andthe H460 tumor region with low expression of PD-L1 has no obvious probe uptake, which proves that the probe has excellent in-vivo targeting performance.
The PET scanning image result shows that the activity of the liver, the kidney and the bladder are very high, particularly the kidney and the bladder, and the sum of the radioactivity of the liver, the kidney and the bladder accounts for more than 70 percent of the total radioactivity of the body after each time point, which indicates that the probe is mainly discharged out of the body through the urinary system. The probe has a rapid clearance half-life (<45min) and18the short half-life period (T1/2-109.8 min) of F is organically combined, so that a high-resolution image can be rapidly and accurately obtained, and the purpose of diagnosing the non-small cell lung cancer is achieved.
The foregoing description is only exemplary of the invention and is not intended to limit the spirit of the invention.
Claims (7)
2. a method for preparing the polypeptide PET molecular imaging probe targeting PD-L1 according to claim 1, characterized by the following steps:
1) dissolving 5-10mg of Cyclopeptide 66 in HEPES-EDTA solution, adding 3-10 equivalents of triisopropylacetamide (2-carboxyethyl) phosphine (dissolved in 0.2 mol/L ammonium acetate buffer solution, pH = 6-7), evacuating the solution, and repeating for three times; reacting at room temperature for 60 minutes, then transferring to a centrifugal filter, and centrifuging for 90 minutes at 4000 revolutions per minute; 1-2 mL of ammonium acetate buffer solution with the concentration of 0.2 mol/L, adding the reduced polypeptide into 1-2 mL of ammonium acetate buffer solution with the concentration of 0.2 mol/L with the pH =6-7, and transferring to a second reaction vessel; 4-6 mg of NOTA-maleimide, 0.1-1mL of 0.2 mol/L sodium acetate buffer (pH = 6-8), adding the mixture into a second reaction vessel, and reacting at 40-60 ℃ for 3 hours; after the reaction was completed, the reaction mixture was transferred to a centrifugal filter and centrifuged at 4000 rpm for 90 minutes; after discarding the flow, adding 2 mL of ultrapure water, centrifuging for 90 minutes again at 4000 rpm, and discarding the flow again; collecting the purified NOTA-PA2881 by using 1mL of ultrapure water, freeze-drying, and obtaining a product after mass spectrometry; the bifunctional chelating agents include, but are not limited to, NOTA, DOTA or NODAGA;
2) labeling the product obtained in the step 1) with radionuclide;
the radionuclides include, but are not limited to18F、68Ga、64Cu or89Zr;
The method for labeling the radionuclide comprises the following steps: mu.L of acetonitrile was added with 5. mu.L of 2mM AlCl30.1M aqueous sodium acetate solution and 2. mu.L of 10 mM DMSO or deionized water, and finally adding Na of 1.85-3.70 GBq (50-100 mCi)18And (3) carrying out high-temperature closed reaction for about 10-20min by using 400 mu L of F solution, and cooling to generate the radionuclide-labeled cyclic peptide complex.
3. The method for preparing a polypeptide PET molecular imaging probe targeting PD-L1 according to claim 2, wherein the radionuclide is18F。
4. The method for preparing the polypeptide PET molecular imaging probe targeting PD-L1 according to claim 2, characterized in that the bifunctional chelating agent is NOTA.
5. The method for preparing a polypeptide PET molecular imaging probe targeting PD-L1 according to claim 2, characterized in that the method for labeling with radionuclides is: dissolving the product obtained after the Cyclopeptide 66 is connected with NOTA in DMSO or acetonitrile for 10 mM, 2-4 mu L; adding AlCl32mM, 10-20 μ L of sodium acetate; adding 200 and 400 mu L acetonitrile; adding QMA purified Na18Solution F, 200-; heating to 100 deg.C under sealed condition, reacting for 10-20min, cooling, passing the reaction solution through pre-activated C18-light solid phase extraction column, washing with deionized water to remove unreacted free substance18F ion, and then 75% ethanol solutionEluting the product from C18-light solid phase extraction column to obtain the product with structural formula18F, diluting the ethanol concentration to be below 10% to prepare the tracer injection.
6. The method for preparing the polypeptide PET molecular imaging probe targeting PD-L1 according to claim 1, characterized in that the tracer agent18The F-AlF-NOTA-PA2881 is used as a PD-L1 molecular targeted tracer in PET molecular imaging.
7. The method for preparing the PD-L1-targeted polypeptide PET molecular imaging probe according to claim 6, characterized in that the tracer agent18The application of F-AlF-NOTA-PA2881 as a tracer for detecting the PD-L1 expression condition of tumors in solid tumor patients (non-small cell lung cancer, gastric cancer and colorectal cancer), wherein the tracer can be specifically combined with PD-L1 protein on tumor cell membranes and can be qualitatively and quantitatively analyzed through various positron nuclide detection devices.
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CN116162127A (en) * | 2023-02-20 | 2023-05-26 | 中国医学科学院阜外医院 | Granzyme B targeted inhibitor, probe and application |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN110172084A (en) * | 2019-05-24 | 2019-08-27 | 哈尔滨医科大学 | 68Ga marks NODAGA modification c-Met molecular probe and preparation and application |
US20190314531A1 (en) * | 2016-12-23 | 2019-10-17 | The Johns Hopkins University | Tumor and immune cell imaging based on pd-l1 expression |
CN111905112A (en) * | 2020-07-31 | 2020-11-10 | 北京肿瘤医院(北京大学肿瘤医院) | Polypeptide compound targeting PD-L1 and application thereof |
CN113583089A (en) * | 2021-07-09 | 2021-11-02 | 西南医科大学附属医院 | PET imaging agent targeting tumor PD-L1, labeled precursor thereof, preparation method and application |
-
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Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20190314531A1 (en) * | 2016-12-23 | 2019-10-17 | The Johns Hopkins University | Tumor and immune cell imaging based on pd-l1 expression |
CN110172084A (en) * | 2019-05-24 | 2019-08-27 | 哈尔滨医科大学 | 68Ga marks NODAGA modification c-Met molecular probe and preparation and application |
CN111905112A (en) * | 2020-07-31 | 2020-11-10 | 北京肿瘤医院(北京大学肿瘤医院) | Polypeptide compound targeting PD-L1 and application thereof |
CN113583089A (en) * | 2021-07-09 | 2021-11-02 | 西南医科大学附属医院 | PET imaging agent targeting tumor PD-L1, labeled precursor thereof, preparation method and application |
Non-Patent Citations (2)
Title |
---|
HAIXIA SUN等: "Design and Discovery of Natural Cyclopeptide Skeleton Based Programmed Death Ligand 1 Inhibitor as Immune Modulator for Cancer Therapy" * |
RAVINDRA A等: "Peptide-Based 68Ga-PET Radiotracer for Imaging PD-L1 Expression in Cancer" * |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN116162127A (en) * | 2023-02-20 | 2023-05-26 | 中国医学科学院阜外医院 | Granzyme B targeted inhibitor, probe and application |
CN116162127B (en) * | 2023-02-20 | 2023-06-23 | 中国医学科学院阜外医院 | Granzyme B targeted inhibitor, probe and application |
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