RGD polypeptide-containing composition and use thereof

Abstract

An RGD polypeptide-containing composition and use thereof. The composition contains 1-3 parts by weight of a compound of formula I, 50-200 parts by weight of TPPTS, 2-250 parts by weight of Tricine, and a succinate buffer system, and the amount of the succinate buffer system is such that the pH of the composition is 4.0-7.0. The composition has excellent integrity and stability, and can be used for preparing SPECT imaging agents for tumor diagnosis.

Description

A composition containing RGD peptide and its application Technical Field

[0001] This invention relates to the field of medical imaging diagnosis, and more specifically to a composition containing RGD polypeptide. Background Technology

[0002] The sustained growth, invasion, and metastasis of malignant tumors depend on tumor angiogenesis. Integrins are important factors involved in regulating tumor angiogenesis and play a crucial role in tumor angiogenesis. Among integrins, integrin α is the most important. v β3 plays the most important role. Integrin α v β3 is highly expressed in tumor neovascular endothelial cells, but not expressed or expressed at low levels in normal cells or mature blood vessels. Studies have confirmed that ligands containing the RGD (Arg-Gly-Asp, arginine-glycine-aspartic acid) sequence can specifically bind to integrin α. v β3, therefore utilizing RGD and integrin α v RGD-type molecular probes designed based on the specific binding of β3 have been widely studied and applied.

[0003] 99m The complex formed by Tc and RGD peptides acts as integrin α. v β3 receptor imaging agents, used as SPECT tracers, can be used for localization diagnosis of integrin-positive tumors, lesion detection, auxiliary diagnosis of neovascularization density, and efficacy evaluation of anti-tumor angiogenesis therapy.

[0004] 99m Tc drug delivery is generally administered by injection. The formulation can be prepared as an injection solution or a lyophilized powder injection, depending on the stability of its components in aqueous solution. The kit containing RGD peptides is then mixed with freshly eluted technetium... 99m Tc] sodium acid injection solution was reacted to produce 99m When using Tc-RGD polypeptide injection, the composition of the packaging and the formulation of the RGD polypeptide need to be comprehensively considered in relation to its labeling. 99m Due to issues such as Tc labeling rate, interactions between components, industrial production as a commodity, logistics, shelf life stability, etc., developing a stable and reliable pharmaceutical formulation is an urgent problem to be solved.

[0005] Summary of the Invention

[0006] This invention provides a composition containing RGD polypeptide for preparing an imaging agent, comprising 1-3 parts by weight of a compound of formula I, 50-200 parts by weight of TPPTS, 2-250 parts by weight of Tricine, and a succinate buffer system, wherein the amount of the succinate buffer system is such that the pH of the composition is 4.0-7.0, and wherein the compound of formula I (abbreviated as RGD) has the following structure:

[0007] The molecular formula of compound I is C 105 H 158 N 25 NaO 35 S has a molecular weight of 2385.58.

[0008] In some embodiments, the composition contains 1 part by weight of the compound of formula I, 50-150 parts by weight of TPPTS, 25-100 parts by weight of Tricine, and a succinate buffer system, wherein the amount of the succinate buffer system is such that the pH of the composition is 5.0-7.0.

[0009] In some embodiments, the composition contains 1-2.5 parts by weight of the compound of formula I, 50-180 parts by weight of TPPTS, 3-220 parts by weight of Tricine, and a succinate buffer system, wherein the amount of the succinate buffer system is such that the pH of the composition is 4.0-6.0.

[0010] In some embodiments, the composition contains 1 part by weight of the compound of formula I, 50-100 parts by weight of TPPTS, 25-75 parts by weight of Tricine, and a succinate buffer system, wherein the amount of the succinate buffer system is such that the pH of the composition is 5.0-6.0.

[0011] In some embodiments, the composition contains 1 part by weight of the compound of formula I, 75 parts by weight of TPPTS, 50 parts by weight of Tricine, and a succinate buffer system, wherein the amount of the succinate buffer system is such that the pH of the composition is adjusted to 5.2-5.6.

[0012] In some embodiments, the succinate buffer system is a succinate buffer system composed of succinic acid and NaOH.

[0013] In some embodiments, the composition contains 40 μg of the compound of formula I, 3 mg TPPTS, 2 mg Tricine, 29.55 mg succinic acid and 17 mg NaOH per unit dose, and the composition has a pH of 5.2-5.6.

[0014] In some embodiments, the composition further contains a freeze-drying protectant, such as mannitol, sorbitol, glycerol, or sucrose, preferably mannitol.

[0015] In some embodiments, the composition is a solid dosage form.

[0016] In some embodiments, the composition is a lyophilized formulation, preferably a lyophilized powder for injection, such as a sterile powder for injection.

[0017] The composition described in this invention has significantly improved stability and can extend the product shelf life.

[0018] In some embodiments, the content of compound I in the composition varies within 5%, 4%, 3%, 2%, 1% or 0.5% within 3 months under conditions of 2-8°C, 40°C±2°C / 75%±5%RH and / or 25°C±2°C / 60%±10%RH.

[0019] In some embodiments, the TPPTS content in the composition varies within 5%, 4%, 3%, 2%, 1%, or 0.5% over 3 months under conditions of 2-8°C, 40°C±2°C / 75%±5%RH, and / or 25°C±2°C / 60%±10%RH.

[0020] In some embodiments, the Tricine content in the composition varies within 5%, 4%, 3%, 2%, 1%, or 0.5% over 3 months under conditions of 2-8°C, 40°C±2°C / 75%±5%RH, and / or 25°C±2°C / 60%±10%RH.

[0021] In another aspect, the present invention provides a liquid formulation or reconstituted liquid formulation, which is obtained by dissolving the composition described in any one of the preceding claims in Na... 99m It was prepared by TcO4 injection.

[0022] In some embodiments, the liquid formulation or reconstituted liquid formulation contains 99m The activity of Tc is 20-50 mCi (preferably 20-35 mCi).

[0023] In some embodiments, the unit dose of the liquid formulation or reconstituted liquid formulation is 0.5-1.5 mL (e.g., 1 mL).

[0024] In some embodiments, the compound of formula I, TPPTS, and Tricine in the liquid formulation or reconstituted liquid formulation are... 99m Tc coordinates to form metal complexes.

[0025] In some embodiments, the chemical structural formula of the metal complex is shown below:

[0026] The molecular formula of the metal complex is C. 122 H 175 N 26 Na3O 46 PS3 99m Tc has a molecular weight of 3036.91.

[0027] In some embodiments, 0.5–1.5 mL of sodium pertechnetate [99mTc] injection at 20–50 mCi (740–1850 MBq), preferably 20–35 mCi, is added to the composition, and the mixture is shaken thoroughly for 1–2 minutes to dissolve. The bottle is then placed in a boiling water bath and heated for 20–30 minutes, and cooled to room temperature to obtain the liquid formulation or reconstituted liquid formulation.

[0028] In some embodiments, the liquid formulation or reconstituted liquid formulation is an injectable.

[0029] In some embodiments, under conditions of 40±2°C and 75%±5% relative humidity, the radiochemical purity of the liquid formulation or reconstituted liquid formulation is above 90%, above 90%, above 91%, above 92%, above 93%, above 94%, above 95%, above 96%, or above 97% within 6 hours.

[0030] In some embodiments, the liquid formulation or reconstituted liquid formulation, under conditions of 25±2°C and 60%±10% relative humidity, has a radiochemical purity of 90% or higher, 90% or higher, 91% or higher, 92% or higher, 93% or higher, 94% or higher, 95% or higher, 96% or higher, or 97% or higher within 6 hours.

[0031] In some implementations, the liquid formulation or reconstituted liquid formulation is used for SPECT / CT imaging.

[0032] In another aspect, the present invention provides a pillbox product comprising the composition described in any one of the preceding claims, and optionally Na. 99m TcO4 injection solution and / or instructions.

[0033] In some embodiments, the composition and Na 99m TcO4 injection is placed in a separate formulation unit.

[0034] In some implementations, the Na 99m The unit dose of TcO4 injection is 0.5-1.5 mL (e.g., 1 mL).

[0035] The SPECT imaging agent prepared from the composition, liquid formulation, or reconstructed liquid formulation described in this invention has shown high tumor targeting in in vivo experiments, specifically recognizing integrin α. v β3-overexpressing tumors. Therefore, in another aspect, the present invention provides the use of the compositions, liquid formulations or reconstituted liquid formulations or kit products described in any of the preceding claims in the preparation of imaging agents, particularly SPECT imaging agents.

[0036] In some embodiments, the imaging agent is used for tumors (e.g., integrin α). v Diagnosis of tumors with high β3 expression, such as localization diagnosis, lesion detection, auxiliary diagnosis of neovascularization density, or evaluation of the efficacy of anti-tumor angiogenesis therapy.

[0037] In some implementations, the imaging agent is used for the diagnosis of lung cancer lymph node metastasis or for the differential diagnosis of benign or malignant lung tumors.

[0038] In another aspect, the present invention provides a diagnostic method comprising the step of administering to a subject an effective amount of any of the preceding compositions, liquid formulations, or reconstituted liquid formulations or kit products, wherein the method is used for tumors (e.g., integrin α). v It can be used for the localization diagnosis of tumors with high β3 expression, lesion detection, auxiliary diagnosis of neovascularization density, and efficacy of anti-tumor angiogenesis therapy.

[0039] In some implementations, the method is used for the diagnosis of lung cancer lymph node metastasis or for the differential diagnosis of benign or malignant lung tumors.

[0040] Terminology Definition

[0041] Unless otherwise defined below, all technical and scientific terms used herein are intended to have the same meaning as commonly understood by one of ordinary skill in the art. References to technical terms herein refer to techniques commonly understood in the art, including variations or equivalent substitutions of techniques that are obvious to one of ordinary skill in the art. While it is believed that the following terms will be well understood by one of ordinary skill in the art, the following definitions are set forth to better explain the invention.

[0042] As used herein, the term “tris(3-sulfonylphenyl)phosphine sodium salt hydrate (TPPTS)” has the following chemical structural formula:

[0043] Molecular formula: C 18 H 12 Na3O9PS3·4H2O; Molecular weight: 640.49; Appearance: White or off-white crystalline powder.

[0044] As used herein, the term "tris(hydroxymethyl)methylglycine" has the following chemical structural formula:

[0045] Molecular formula: C6H 13 NO5; Molecular weight: 179.17; Appearance: White crystals or crystalline powder.

[0046] As used herein, the term "labeling rate" refers to the percentage of stable radionuclides introduced into a labeled compound molecule relative to the total amount of nuclides used in the labeling reaction; it is a parameter reflecting labeling efficiency. The same radionuclide can have multiple different valence states, thus forming different compounds. To describe the relative quantitative relationships between the various chemical forms of the same radionuclide, the concept of radiochemical purity, also known as labeling rate, is defined. Commonly used methods for determining this include radiometric paper chromatography, thin-layer chromatography, column chromatography, and protein precipitation. Attached Figure Description

[0047] The accompanying drawings, which are included to provide a further understanding of the invention and form part of this application, illustrate exemplary embodiments of the invention and, together with their description, serve to explain the invention and do not constitute an undue limitation thereof. In the drawings:

[0048] Figure 1 shows the radiometric HPLC spectra of different formula I compounds (referred to as RGD) labeled with different amounts.

[0049] Figure 2 shows the dosage box labeling rate of different Formula I compounds (abbreviated as RGD).

[0050] Figure 3 shows the radioactive HPLC spectra labeled with different amounts of TPPTS.

[0051] Figure 4 shows the labeling rate of different TPPTS dosage kits.

[0052] Figure 5 shows the radioactive HPLC spectra labeled with different amounts of Tricine.

[0053] Figure 6 shows the labeling rate of different Tricine dosage kits.

[0054] Figure 7 shows the radiometric HPLC spectra of the kit labels at different pH values.

[0055] Figure 8 shows the labeling rate of the medicine box at different pH levels.

[0056] Figure 9 shows the administration of the drug via tail vein injection in a lung cancer tumor model mouse. 99m Distribution of radioactivity in different tissues at different time points after administration of Tc-RGD peptide (8 μg / kg) (ng / g tissue).

[0057] Figure 10 shows 99mSPECT / CT imaging of Tc-RGD polypeptide injection in mice bearing NCI-H1975 lung cancer NOD SCID, with the arrow indicating the tumor location.

[0058] Figure 11 shows 99m SPECT images of Tc-RGD peptide injection 2 hours after injection into NOD SCID mice carrying NCI-H1975 lung cancer. The left image is the unblocked group, and the right image is the pre-blocked group with excessive RGD peptide.

[0059] Figure 12 shows 99m SPECT imaging of Tc-RGD peptide injection in NOD SCID mice bearing bilateral NCI-H1975 / NCI-H157 tumors. The lower arrow indicates the location of the NCI-H1975 tumor, and the upper arrow indicates the location of the NCI-H157 tumor.

[0060] Figure 13 shows 99m SPECT images 2 hours after injection of Tc-RGD peptide into NOD SCID mice bearing bilateral NCI-H1975 / NCI-H157 tumors. The left image shows no blocking, while the right image shows pre-blocking with excess RGD peptide. The lower arrow indicates the location of the NCI-H1975 tumor, and the upper arrow indicates the location of the NCI-H157 tumor. Detailed Implementation

[0061] The embodiments of the present invention will be described in detail below with reference to examples. However, those skilled in the art will understand that the following examples are for illustrative purposes only and should not be considered as limiting the scope of the invention. Unless otherwise specified in the examples, conventional conditions or conditions recommended by the manufacturer are followed. Reagents or instruments whose manufacturers are not specified are all commercially available conventional products.

[0062] Example 1: Kit Formulation Screening Experiment for RGD Peptides

[0063] I. Experimental Objective:

[0064] The effects of different concentrations of Formula I compound, TPPTS, tricine, and pH on the labeling rate of the medicine box were investigated to screen prescriptions.

[0065] II. Experimental Apparatus:

[0066] Instrument name and model

[0067] Water bath (domestic); activity meter (CAPINTEC CRC-15R); HPLC instrument (Agilent 1260 high performance liquid chromatograph).

[0068] Chromatographic conditions

[0069] HPLC instrument: Agilent 1260 high performance liquid chromatograph;

[0070] Column: COSMOSIL 5C 18 -MS-II, 4.6ID×250mm, 5μm;

[0071] Preparation of mobile phase: Phase A (dissolve 6.8g of potassium dihydrogen phosphate and 20.0g of sodium heptanesulfonate in 800mL of water, add 7.0mL of triethylamine, adjust the pH to 3.0 with phosphoric acid, dilute with water to 1000mL, and filter to obtain the mobile phase); Phase B is acetonitrile, with a flow rate of 1mL / min.

[0072] Detection wavelength: 210nm, radioactivity detector Channel A;

[0073] rinsing gradient:

[0074] Column temperature: 40℃;

[0075] Injection volume: 20 μL;

[0076] Diluent: physiological saline.

[0077] III. Process and Results:

[0078] 1) Selection of the amount of compound I

[0079] The dosage of this compound of formula I is based on... 99m Tc is used to determine the radiochemical purity of the labeling reaction of compound I.

[0080] Experimental description: 99m When Tc-labeling compounds of formula I, the concentration of formula I significantly affects the radioactive labeling rate. Insufficient concentration of formula I compound leads to a low labeling rate, thus impacting the final labeling outcome. 99m The radiochemical purity of Tc-3PRGD2 was determined. In this experiment, different amounts of Formula I compounds were used in the labeling process, and the labeling rate under various conditions was detected by radioactive HPLC. Based on the hospital-prepared formulation at pH 4.8, the optimal amount of Formula I compound for this labeling process was screened. Detailed data are shown in Table 1 and Figures 1-2.

[0081] Table 1 Experiment on the selection of dosage of ligand I compounds

[0082] The results of this experiment show that when the content of compound I is between 30 and 70 μg, its labeling rate reaches over 90%. Considering that compound I is a highly active biological product and a ligand in the kit components, it may produce degradation impurities during long-term storage. Therefore, to ensure that this product can be used normally throughout its shelf life, the dosage is tentatively set at 40 μg / vial.

[0083] 2) Selection of TPPTS dosage

[0084] Experimental Description: The amount of compound I was 40 μg, while the amounts of other components remained unchanged, to prepare a kit without TPPTS. In subsequent radiolabeling experiments, different masses of TPPTS were added to the above kit, and then technetium […]. 99m Add 740–1300 MBq (20–35 mCi) of TPTTS sodium injection solution to the vial until the final volume is 1 mL, shake thoroughly to dissolve the solids in the vial. Place the vial in a 100°C water bath for 20 min, then remove and cool to room temperature to obtain technetium [TPTTS] solutions with different TPTTS contents. 99m The labeling rate of sodium toluenesulfonate nicotinamide polyethylene glycol bicyclic RGD peptide injection was investigated using radioactive HPLC. The experimental results are shown in Tables 2-3 and Figures 3-4.

[0085] Table 2 Prescriptions without TPPTS kits (single dose)

[0086] Table 3. Screening of TPPTS Dosage

[0087] With other ligand dosages remaining constant, when the TPPTS dosage is between 0 and 1.5 mg / vial, the radiolabeling rate of the kit increases with the increase of TPPTS dosage, showing a positive correlation. When the TPPTS dosage is in the range of 1.5 to 5 mg / vial, the radiolabeling rate of the kit does not change significantly, and the labeling rate can be guaranteed to reach more than 90%.

[0088] Because TPPTS exhibits both ligand and reducing agent properties in this system, and considering that the total TPPTS content may decrease due to factors such as oxidation during long-term storage, the dosage of trisodium tri(3-sulfonylphenyl)phosphine is set at 3 mg / vial to ensure normal use throughout the product's shelf life.

[0089] 3) Selection of Tricine dosage

[0090] Experimental instructions: The amount of compound I was 40 μg, and the amount of TPPTS was 3 mg, to prepare a tricine-free kit. In the subsequent radiolabeling experiment, different masses of tricine were added to the above kit, and then 1 mL of technetium […]. 99mAdd 740–1300 MBq (20–35 mCi) of sodium tricine injection to the vial and shake thoroughly to dissolve the solids in the vial. Place the vial in a 100°C water bath for 20 minutes, then remove and cool to room temperature to obtain technetium [tricine] with different triciline contents. 99m The labeling rate of sodium toluenesulfonate nicotinamide polyethylene glycol bicyclic RGD peptide injection was investigated using radioactive HPLC. The experimental results are shown in Tables 4-5 and Figures 5-6.

[0091] Table 4. Prescriptions without Tricine (single dose)

[0092] Table 5. Tricine Dosage Screening

[0093] (1) When the dosage of other ligands remains unchanged, when the dosage of Tricine is 0 to 0.3 mg / vial, the radiolabeling rate of the kit increases with the increase of Tricine dosage, showing a positive correlation; when the dosage of Tricine is in the range of 0.3 mg to 6.5 mg / vial, the radiolabeling rate of the kit does not change significantly, and the labeling rate can be guaranteed to reach more than 90%.

[0094] (2) Tricine is a marker for HYNIC as a bifunctional chelating agent. 99m One of the universal ligands of Tc, its main function is to stabilize newly generated [ 99m Tc-HYNIC] core, reducing technetium [ 99m The generation of Tc colloids improves the labeling rate and increases the hydrophilicity of the labeled substance. When used as an in-hospital preparation, the prescribed dosage of Tricine is 6.5 mg. To ensure a more stable radiolabeling rate with the new prescription cartridge, and considering the ratio of Tricine to TPPTS as a co-ligand, the dosage of Tricine is set at 2 mg / vial.

[0095] 4) pH value selection

[0096] Experimental instructions: Use kits with different pH values. In the subsequent radiolabeling experiment, 1 mL of technetium [ 99m Add 740–1300 MBq (20–35 mCi) of Tc sodium injection to the bottle and shake thoroughly to dissolve the solids in the container. Place the container in a 100°C water bath for 20 minutes, then remove and cool to room temperature to obtain technetium [Tc] at different pH values. 99m The labeling rate of sodium toluenesulfonate nicotinamide polyethylene glycol bicyclic RGD peptide injection was investigated using radioactive HPLC. The experimental results are shown in Tables 6-7 and Figures 7-8.

[0097] Table 6. Prescriptions for medicine boxes at different pH levels

[0098] Table 7 Screening at different pH levels

[0099] (1) With the dosage of the active ingredient and other ligands remaining constant, the radiolabeling rate of the kit increases with the decrease of pH between 4.5 and 6.5, showing a negative correlation; when the pH is in the range of 4.5 to 5.8, the radiolabeling rate of the kit does not change significantly, and the labeling rate can be guaranteed to reach more than 90%.

[0100] (2) Succinate buffer provides an acidic labeling environment, improving the labeling rate. When this product is used as an in-house preparation, the prescription pH is 4.8. To ensure longer shelf-life stability of the new prescription kit, the pH is set at 5.5.

[0101] Experimental conclusions

[0102] The prescription on the medicine box is:

[0103] *Mannitol is used as a freeze-drying adsorbent and does not participate in the labeling process.

[0104] Example 2 Stability Study

[0105] This embodiment describes the RGD peptide's kit and 99m The stability of Tc-RGD peptide injection was investigated. 99m Tc-RGD peptide injection is a locally labeled radiodiagnostic agent, consisting of a radiochemical reagent and a kit containing a compound of Formula I. Based on the formulation screening experiment in Example 1, the kit composition of the radiochemical reagent and the compound of Formula I was determined, as detailed in Table 8.

[0106] 99m Preparation method of Tc-RGD polypeptide injection: Take one vial of compound I, and under aseptic conditions, add 1 mL of technetium [ 99m Add 20–35 mCi (740–1300 MBq) of sodium thiocyanate injection to the bottle and shake thoroughly to dissolve the solids in the medicine box. Place the medicine box in a 100°C water bath for 20 minutes, then remove and cool to room temperature to obtain the final product.

[0107] Table 8 Preparation 99m Tc-RGD polypeptide injection kit product

[0108] (1) Stability under different pH conditions

[0109] According to the prescription in Table 8, add approximately half the prescribed amount (8 mg) of sodium hydroxide. After dissolving, add the prescribed amount of succinic acid. After the succinic acid is completely dissolved, add TPPTS and Tricine. Then, adjust the pH of the solution to the required pH value with 1 mol / L sodium hydroxide solution. Prepare a concentrated solution of Formula I compound separately, transfer it to the above-prepared solution according to the prescribed amount, and make up to the full volume. Then, place it at 15℃ and 25℃ respectively to detect the changes in the content of Formula I compound and related substances. See Table 9 below for details. The above-prepared drug compositions with different pH values ​​were lyophilized, reconstituted, and the pH value was detected. 740-1300 MBq of sodium pertechnetium [99mTc]ate injection solution was added and mixed to react. The labeling rate under various pH conditions was detected by radioactive HPLC. See Table 10 below for details.

[0110] Table 9. Stability of the drug solution under different pH conditions

[0111] Table 10. Marking rate of freeze-dried medicine boxes at different pH values

[0112] As can be seen from Tables 8-10 above, within the pH range of the present invention, especially within the pH range of 5.2-5.6, the content of the compound of Formula I and its total impurities do not change significantly with time, and the labeling rate can reach 95%.

[0113] (2) Chemical stability

[0114] This experiment investigated the long-term stability of the compound of formula I in the chromatographic kit under the following conditions: 40℃±2℃ / 75%±5%RH. The content was determined by HPLC using octadecylsilane-bonded silica gel (250×4.6mm, 5μm) as the packing material. The results are shown in Table 11.

[0115] Table 11 shows the stability data of the kit for Formula I compound under conditions of 40℃±2℃ / 75%±5%RH.

[0116] (3) Radiostability

[0117] This experiment investigated 99m The radiostability of Tc-RGD polypeptide injection was determined by radiometric HPLC, and the labeling rate was detected. The results are shown in Tables 12 and 13.

[0118] Table 12 Stability data under conditions of 40±2℃ and 75%±5% relative humidity.

[0119] Table 13 Stability data under conditions of 25±2℃ and 60%±10% relative humidity

[0120] The above data indicate that the kit for the compound of formula I of this invention and high technetium [ 99m The sodium tc] solution was prepared by sodium tc] acid injection. 99m The Tc-RGD polypeptide injection met the stability requirements within 6 hours under conditions of 40±2℃ and 75%±5% relative humidity, and 25±2℃ and 60%±10% relative humidity.

[0121] Example 3 Pharmacodynamic Experiment

[0122] (1) Tissue distribution assay: Thirty NOD SCID mice (15 males and 15 females) bearing NCI-H1975 lung cancer with good tumor growth were divided into 5 groups (4 normal distribution groups and 1 cold peptide blocking group). The cold peptide blocking group was injected intraperitoneally with a saline solution of compound I 0.5 hours prior to administration. The injection was administered via the tail vein. 99m Animals were sacrificed at different time points after Tc-RGD polypeptide injection (the cold peptide blocking group was sacrificed at 2 hours), and plasma and tissues were collected.

[0123] (2) Imaging test:

[0124] 1. Two NOD SCID mice bearing NCI-H1975 lung cancer with good tumor growth status were injected via tail vein. 99m In vivo SPECT / CT imaging was performed after Tc-RGD polypeptide injection.

[0125] 2. Two mice bearing NCI-H1975 / NCI-H157 bilateral tumor models with good bilateral tumor growth were used for further analysis. 99m In vivo SPECT imaging of Tc-RGD peptides.

[0126] Dosage: Chemical dose 8 μg·kg -1 Radiation dose 107.6 MBq·kg -1 .

[0127] The results are shown in Table 14 and Figures 9-13.

[0128] Table 14. Drug administration via tail vein injection in lung cancer tumor model mice. 99m Distribution of radioactivity in different tissues at different time points after administration of Tc-RGD peptide (8 μg / kg) (ng / g tissue) Note: ND: Not detected; Mean±SD, n=6

[0129] Tissue distribution assays showed that the radiopharmaceutical content was high in the tumors of the model mice, while the drug penetration into the brain tissue was extremely low, indicating that the imaging agent of this invention has significant tumor targeting properties. In the control group, i.e., the cold peptide blocking group, very low radioactivity was detected in all tissues, indicating that the cold peptide blocking group can competitively inhibit... 99m Tc injection and integrin αv β3 receptor binding. Imaging results further confirmed that this drug targets integrin α. v Specificity of β3 receptor.

[0130] Although specific embodiments of the invention have been described in detail, those skilled in the art will understand that various modifications and substitutions can be made to those details based on all the teachings disclosed, and all such changes are within the scope of protection of the invention. The full scope of the invention is given by the appended claims and any equivalents thereof.

Claims

1. A composition comprising 1-3 parts by weight of a compound of formula I, 50-200 parts by weight of TPPTS, 2-250 parts by weight of Tricine and a succinate buffer system, wherein, The amount of the succinate buffer system is such that the pH of the composition is between 4.0 and 7.0, wherein the compound of formula I has the following structure:

2. The composition of claim 1, comprising 1-2.5 parts by weight of the compound of formula I, 50-180 parts by weight of TPPTS, 3-220 parts by weight of Tricine and a succinate buffer system, wherein, The amount of the succinate buffer system used is such that the pH of the composition is adjusted to 4.0-6.0; Preferably, the composition contains 1 part by weight of the compound of formula I, 50-150 parts by weight of TPPTS, and 25- 100 parts by weight of Tricine and succinate buffer system, wherein the amount of succinate buffer system is such that the pH of the composition is adjusted to 5.0-7.0; Preferably, the composition contains 1 part by weight of the compound of formula I, 50-100 parts by weight of TPPTS, 25-75 parts by weight of Tricine and a succinate buffer system, wherein the amount of the succinate buffer system is such that the pH of the composition is 5.0-6.

0. Preferably, the composition contains 1 part by weight of the compound of formula I, 75 parts by weight of TPPTS, 50 parts by weight of Tricine and a succinate buffer system, wherein the amount of the succinate buffer system is such that the pH of the composition is 5.2-5.

6. Preferably, the succinate buffer system is a succinate buffer system composed of succinic acid and NaOH; Preferably, the composition contains 40 μg of the compound of formula I, 3 mg TPPTS, 2 mg Tricine, 29.55 mg succinic acid and 17 mg NaOH per unit dose, and the pH of the composition is 5.2-5.

6.

3. The composition of claim 1 or 2 further comprises a freeze-drying protectant, such as mannitol, sorbitol, glycerol or sucrose, preferably mannitol; Preferably, the composition is a solid dosage form; Preferably, the composition is a lyophilized preparation, more preferably a lyophilized powder for injection, such as a sterile powder for injection.

4. The composition according to any one of claims 1-3, under conditions of 40℃±2℃ and 75%±5%RH for 3 months, the content of the compound of formula I changes within 5%, 4%, 3%, 2%, 1%, or 0.5%. Preferably, under conditions of 40℃±2℃ and 75%±5%RH, the TPPTS content changes within 5%, 4%, 3%, 2%, 1%, or 0.5% within 3 months. Preferably, under conditions of 40℃±2℃ and 75%±5%RH, the Tricine content changes within 5%, 4%, 3%, 2%, 1%, or 0.5% within 3 months.

5. A liquid formulation or reconstituted liquid formulation, wherein the composition according to any one of claims 1-4 is dissolved in Na... 99m It was prepared by TcO4 injection; Preferably, in the liquid formulation or reconstituted liquid formulation 99m The activity of Tc is 20-50 mCi (preferably 20-35 mCi); Preferably, the unit dose of the liquid formulation or reconstituted liquid formulation is 0.5-1.5 mL (e.g., 1 mL); Preferably, the compound of formula I, TPPTS, and Tricine are combined with... 99m Tc forms metal complexes; Preferably, the liquid formulation or reconstituted liquid formulation is an injectable.

6. The liquid formulation or reconstituted liquid formulation according to claim 5, under the conditions of 40±2℃ and 75%±5% relative humidity, has a radiochemical purity of more than 90%, more than 90%, more than 91%, more than 92%, more than 93%, more than 94%, more than 95%, more than 96%, or more than 97% within 6 hours. Preferably, the liquid formulation or reconstituted liquid formulation, under conditions of 25±2℃ and 60%±10% relative humidity, has a radiochemical purity of 90% or higher, 90% or higher, 91% or higher, 92% or higher, 93% or higher, 94% or higher, 95% or higher, 96% or higher, or 97% or higher within 6 hours.

7. The liquid formulation or reconstituted liquid formulation according to claim 5 or 6, for use in SPECT / CT imaging.

8. A pillbox product comprising the composition according to any one of claims 1-4, and optionally Na. 99m TcO4 injection and / or instructions for use; Preferably, the composition and Na 99m TcO4 injection is placed in a separate formulation unit; Preferably, the Na 99m The unit dose of TcO4 injection is 0.5-1.5 mL (e.g., 1 mL).

9. Use of the composition according to any one of claims 1-4, the liquid formulation or reconstituted liquid formulation according to any one of claims 5-7, and the cassette product according to claim 8 in the preparation of imaging agents, particularly SPECT imaging agents; Preferably, the imaging agent is used for tumors (e.g., integrin α). v Diagnosis of tumors with high β3 expression, such as localization diagnosis, lesion detection, auxiliary diagnosis of neovascularization density, or evaluation of the efficacy of anti-tumor angiogenesis therapy; Preferably, the imaging agent is used for the diagnosis of lung cancer lymph node metastasis or for the differential diagnosis of benign and malignant lung tumors.

10. A diagnostic method comprising the step of administering to a subject an effective amount of the composition according to any one of claims 1-4, the liquid formulation or reconstituted liquid formulation according to any one of claims 5-7, or the box product according to claim 8, wherein, The method is used for tumors (e.g., integrin α). v The efficacy of anti-tumor angiogenesis therapy in the localization diagnosis of tumors with high β3 expression, lesion detection, auxiliary diagnosis of neovascularization density, and tumor localization diagnosis. Preferably, the method is used for the diagnosis of lung cancer lymph node metastasis or for the differential diagnosis of benign and malignant lung tumors.

Images