WO2023202730A2 - 放射性伊文思蓝衍生物药物水溶液及其制备方法和用途 - Google Patents

放射性伊文思蓝衍生物药物水溶液及其制备方法和用途 Download PDF

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WO2023202730A2
WO2023202730A2 PCT/CN2023/101452 CN2023101452W WO2023202730A2 WO 2023202730 A2 WO2023202730 A2 WO 2023202730A2 CN 2023101452 W CN2023101452 W CN 2023101452W WO 2023202730 A2 WO2023202730 A2 WO 2023202730A2
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solution
reaction
aqueous solution
stabilizer
formula
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PCT/CN2023/101452
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French (fr)
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WO2023202730A3 (zh
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阳国桂
郝晋
杜泽天
田佳乐
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北京先通国际医药科技股份有限公司
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Priority claimed from CN202210416297.4A external-priority patent/CN114796532A/zh
Priority claimed from CN202210416293.6A external-priority patent/CN114748471A/zh
Application filed by 北京先通国际医药科技股份有限公司 filed Critical 北京先通国际医药科技股份有限公司
Publication of WO2023202730A2 publication Critical patent/WO2023202730A2/zh
Publication of WO2023202730A3 publication Critical patent/WO2023202730A3/zh

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K51/00—Preparations containing radioactive substances for use in therapy or testing in vivo
    • A61K51/12—Preparations containing radioactive substances for use in therapy or testing in vivo characterised by a special physical form, e.g. emulsion, microcapsules, liposomes, characterized by a special physical form, e.g. emulsions, dispersions, microcapsules
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Definitions

  • the present application relates to an aqueous radiopharmaceutical solution with high chemical stability and high radiochemical stability and a preparation method thereof, and in particular to a radionuclide complex modified with an Evans blue (EB) fragment.
  • EB Evans blue
  • the circulating half-life of a drug molecule in the body can greatly affect the effectiveness and availability of the drug.
  • radiopharmaceuticals need a long enough in vivo half-life to allow the radiopharmaceutical to interact with the target. fully bind to the target site while being gradually cleared from other non-target tissues or organs to achieve its intended diagnostic and/or therapeutic effect.
  • HSA human serum albumin
  • Evans blue is an azo dye that is commonly used to detect blood-brain barrier integrity, vascular permeability, blood volume, and cell viability due to its high affinity for serum albumin.
  • the structural fragments of Evans blue dye molecules are used to structurally modify targeting molecules (such as DOTATATE, PSMA-617, etc.), and the resulting Evans blue derivative molecules, such as DOTA-EB-TATE, EB-PSMA, etc., can be used through molecular and Endogenous serum albumin reversibly binds, using serum albumin as a reversible carrier of drug molecules to extend the half-life of drug molecules in the blood. It has been reported in the literature (Bioconjugate Chem.
  • the relative molecular weight of the Evans blue fragment and linker group (Linker) introduced in the drug molecular structure is relatively large, which will greatly affect the physical and chemical properties of the molecule, making aqueous drug solutions containing Evans blue derivatives face difficulties in the formulation process research. More challenges.
  • the active ingredient (API) of the drug PLUVICTO TM (lutetium Lu 177 vipivotide tetraxetan) injection approved for the treatment of PSMA-positive metastatic castration-resistant prostate cancer (mCRPC) is [ 177 Lu]Lu-PSMA-617
  • the stabilizers are 0.39mg/mL gentisic acid and 50.0mg/mL sodium ascorbate.
  • the purpose of this application is to provide an aqueous radiopharmaceutical solution, which contains a complex formed by a targeting molecule modified by an Evans blue (EB) fragment and a radioactive metal nuclide, and a stabilizer. Furthermore, this application also provides a method for preparing the radiopharmaceutical aqueous solution. Specifically, the present invention relates to the following:
  • Aqueous radiopharmaceutical solution which includes: a compound represented by Formula I or a pharmaceutically acceptable ester, amide, solvate, salt thereof, or a salt of a compound represented by Formula I or a pharmaceutically acceptable ester thereof , or a salt of a compound represented by formula I or a pharmaceutically acceptable amide thereof, or a solvate of a compound represented by formula I or a pharmaceutically acceptable ester thereof, or a compound represented by formula I or a salt thereof.
  • the solvate of a pharmaceutically acceptable amide, or the solvate of a compound represented by formula I or a pharmaceutically acceptable salt thereof, a complex formed with a radioactive metal nuclide, and a total concentration of 0.5-400 mg/ mL of stabilizer present
  • L 1 is —(CH 2 ) m —, where m is an integer from 0 to 12, where each CH 2 may individually be replaced by —O—, —NH(CO)—, or —(CO)NH—, condition is that no two adjacent CH 2 groups are replaced;
  • L 3 is —(CH 2 ) n —, where n is an integer from 0 to 12, where each CH 2 may individually be replaced with —O—, —NH(CO)—, or —(CO)NH—, condition is that no two adjacent CH 2 groups are replaced;
  • Ch is a chelating group
  • Tg is the targeting group.
  • Tg is selected from the group consisting of agents capable of targeting somatostatin receptor (SSTR), prostate-specific membrane antigen (PSMA), fibroblast activation protein (FAP), folate receptor (FR), epidermal growth factor receptor or integrin compound group.
  • SSTR somatostatin receptor
  • PSMA prostate-specific membrane antigen
  • FAP fibroblast activation protein
  • FR folate receptor
  • epidermal growth factor receptor or integrin compound group is selected from the group consisting of agents capable of targeting somatostatin receptor (SSTR), prostate-specific membrane antigen (PSMA), fibroblast activation protein (FAP), folate receptor ( FR), epidermal growth factor receptor or integrin compound group.
  • radioactive metal nuclide is selected from 177 Lu, 99m Tc, 68 Ga, 64 Cu, 67 Cu, 111 In, 86 Y, 90 Y, 89 Zr, 186 Re , 188 Re, 153 Sm, 82 Rb, 166 Ho, 225 Ac, 212 Pb, 213 Bi, 212 Bi or 227 Th.
  • the stabilizer is selected from the group consisting of gentisic acid and its salts, ascorbic acid and its salts, histidine, cysteine and its salts, methyl One or more of sulfate, selenomethionine, thiosulfate, maltose, inositol, benzyl alcohol, trehalose, povidone, nicotinamide, ethanol, curcumin, and melatonin, Preferably, it is one or more of gentisic acid, ethanol, and methionine.
  • the pharmaceutical aqueous solution according to item 9, wherein the stabilizer added during the reaction to form the complex is selected from the group consisting of gentisic acid and its salts, ascorbic acid and its salts, histidine, cysteine and One of its salts, methionine, selenomethionine, thiosulfate, maltose, inositol, benzyl alcohol, trehalose, povidone, nicotinamide, ethanol, curcumin, melatonin, or Two or more types, preferably gentisic acid;
  • the stabilizer added after the reaction to form the complex is selected from the group consisting of gentisic acid and its salts, ascorbic acid and its salts, histidine, cysteine and its salts, methionine, and selenium methionine.
  • gentisic acid and its salts ascorbic acid and its salts
  • histidine cysteine and its salts
  • methionine methionine
  • selenium methionine preferably gentisic acid, ethanol, or Methionine.
  • the pharmaceutical aqueous solution according to item 10, wherein the stabilizers added during the reaction to form the complex and after the reaction are completed are selected from gentisic acid, and the gentisic acid is in the pharmaceutical aqueous solution.
  • the gentisic acid is in the pharmaceutical aqueous solution.
  • gentisic acid is present in the aqueous pharmaceutical solution at a concentration of 0.1-10 mg/mL, preferably 0.5-5 mg/mL, and ethanol is present at a concentration of 0-400 mg/mL.
  • the pharmaceutical aqueous solution according to item 10, wherein the stabilizer added during the reaction to form the complex is gentisic acid, and the stabilizer added after the reaction to form the complex is completed is methyl sulfide.
  • Acid, and gentisic acid is present in the concentration of 0.1-10 mg/mL, preferably 0.5-5 mg/mL, and methionine is present in the aqueous pharmaceutical solution at a concentration of 0-50 mg/mL.
  • the pharmaceutical aqueous solution according to item 10 which further includes a buffer, which may be selected from acetate, citrate, phosphate or formate solutions; the buffer salt in the buffer is in the pharmaceutical
  • concentration in aqueous solution is 0.005-0.5M.
  • the pharmaceutical aqueous solution according to item 10 which further includes a cosolvent selected from the group consisting of polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 80, poloxamer 188, polysorbate One or two of oxyethylene castor oil, Span, ethanol, propylene glycol, glycerol, polyethylene glycol (average molecular weight 200 to 8000), sorbitol, dimethyl sulfoxide, and sodium lauryl sulfate More than one species, polysorbate 80 is preferred.
  • the aqueous pharmaceutical solution according to item 10 which further includes a free nuclide chelating agent, which can be selected from pentetic acid and its salts; the free nuclide chelating agent is in the aqueous pharmaceutical solution. Present at a concentration of 0.005-0.1mg/mL.
  • a method for preparing an aqueous radiopharmaceutical solution comprising a complex formed by a radionuclide and an Evans blue derivative molecule, characterized by comprising the following steps:
  • the reaction vessel After a given time, add the solution containing the Evans blue derivative molecules to the reaction vessel.
  • the given time is 0.1 to 20 minutes, and more preferably 3 to 10 minutes;
  • the Evans blue derivative molecule reacts with a radionuclide to obtain the radionuclide complex
  • the Evans blue derivative molecule is a compound represented by Formula I or a pharmaceutically acceptable ester, amide, solvate, salt thereof, or a compound represented by Formula I or a pharmaceutically acceptable ester thereof
  • L 1 is —(CH 2 ) m —, where m is an integer from 0 to 12, where each CH 2 may individually be replaced by —O—, —NH(CO)—, or —(CO)NH—, condition is that no two adjacent CH 2 groups are replaced;
  • L 3 is —(CH 2 ) n —, where n is an integer from 0 to 12, where each CH 2 may individually be replaced with —O—, —NH(CO)—, or —(CO)NH—, condition is that no two adjacent CH 2 groups are replaced;
  • Ch is a chelating group
  • Tg is the targeting group.
  • radioactive nuclide is selected from 177 Lu, 99m Tc, 68 Ga, 64 Cu, 67 Cu, 111 In, 86 Y, 90 Y, 89 Zr, 186 Re , 188 Re, 153 Sm, 82 Rb, 166 Ho, 225 Ac, 212 Pb, 213 Bi, 212 Bi or 227 Th.
  • Ch in formula I is selected from Preferably
  • Tg in formula I is selected from the group consisting of somatostatin receptor (SSTR), prostate-specific membrane antigen (PSMA), and fibroblast activation protein (FAP). , folate receptor (FR), epidermal growth factor receptor or integrin compound group.
  • the raw material bottle is rinsed with a rinse solution, and the rinsed solution is transferred into the reaction container and mixed with the solution containing radionuclides.
  • the flushing liquid is an aqueous solution, preferably selected from a solution containing a first stabilizer, a solution containing buffer salt, water or sodium chloride injection; more preferably , repeat rinsing one or more times with rinsing solution.
  • the method according to item 19 characterized in that, in the step of reacting the Evans blue derivative molecule with the radionuclide, the molar ratio between the Evans blue derivative molecule and the radionuclide is is 1.5-50, preferably 5-20.
  • the reaction temperature is 50-100°C, preferably 60-80°C, and the reaction time It is 5-60 minutes, preferably 10-30 minutes.
  • the first stabilizer is selected from the group consisting of gentisic acid and its salts, ascorbic acid and its salts, histidine, cysteine and its salts, methionine , one or more of selenomethionine, thiosulfate, maltose, inositol, benzyl alcohol, trehalose, povidone, nicotinamide, ethanol, curcumin, and melatonin, preferably dragon cholic acid.
  • the second stabilizer is selected from the group consisting of gentisic acid and its salts, ascorbic acid and its salts, histidine, cysteine and its salts, methionine , one or more of selenomethionine, thiosulfate, maltose, inositol, benzyl alcohol, trehalose, povidone, nicotinamide, ethanol, curcumin, and melatonin, preferably dragon Cholic acid, ethanol or methionine.
  • a buffer salt solution is added before the Evans blue derivative molecule reacts with the radionuclide.
  • the buffer salt solution is present in the first compound containing stabilizer solution.
  • the buffer salt solution is selected from acetate, citrate, phosphate or formate solutions, preferably an acetic acid-sodium acetate buffer salt solution.
  • step of adding the solution containing the second stabilizer to the reaction vessel after a given reaction time further includes adding a co-solvent to the reaction vessel.
  • the co-solvent is selected from the group consisting of polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 80, poloxamer 188, polyoxyethylene castor One or more of sesame oil, Span, ethanol, propylene glycol, glycerin, polyethylene glycol (average molecular weight 200-8000), sorbitol, dimethyl sulfoxide, sodium lauryl sulfate, Polysorbate 80 is preferred.
  • the step of adding a solution containing a second stabilizer to the reaction vessel after a given reaction time further includes adding a free nuclide chelating agent to the reaction vessel.
  • the chelating agent can be selected from pentetic acid and its salts, preferably pentetic acid.
  • Radiopharmaceutical aqueous solution prepared by the method described in any one of items 19 to 38.
  • aqueous pharmaceutical solution according to item 39 which is the aqueous radiopharmaceutical solution according to any one of items 1 to 17.
  • a radioactive treatment method for tumors which includes administering the aqueous pharmaceutical solution according to any one of items 1 to 17 to a subject in need.
  • the stabilizer can be a single one, or a combination of two or more. In some embodiments, it is preferred to use only gentisic acid or gentisic acid and ethanol. Ethanol has good Resistance to radiolysis; at the same time, the stabilizer preferably does not use ascorbic acid. The addition of ascorbic acid is not conducive to the stability of Evans blue derivatives.
  • pentetic acid is added as a chelating agent after the reaction to chelate unreacted free nuclide ions to reduce the impact of free radionuclide ions on healthy tissues in the body. Produce unnecessary exposure.
  • the initial radiochemical purity after preparation of the radiopharmaceutical is not less than 93%, preferably not less than 95%, and more preferably not less than 97%.
  • the radiochemical purity of the API of the Evans blue derivative drug aqueous solution using the solution of the present application is maintained at no less than 90% within 48 hours under storage conditions of 32°C and 60% RH, preferably , remains no less than 90% within 72 hours.
  • the concentration of gentisic acid in the reaction phase solution is controlled to be 0.6-20.0 mg/mL.
  • the concentration of gentisic acid in the reaction phase solution is controlled to be 0.6-20.0 mg/mL.
  • the concentration of gentisic acid in the reaction phase solution is controlled to be 0.6-20.0 mg/mL.
  • the anti-radiodecomposition effect of gentisic acid is insufficient.
  • it is higher than 20.0mg/mL the high concentration of gentisic acid will delay the reaction kinetics and adversely extend the time required for the reaction.
  • This control range is to reduce the concentration of gentisic acid as much as possible to avoid adverse effects on the reaction kinetics while ensuring the stability of the solution.
  • the solution containing the Evans blue derivative molecule and the nuclide solution Before mixing the solution containing the Evans blue derivative molecule and the nuclide solution, first mix the nuclide solution with the solution containing the first stabilizer, and after a given time, add the Evans blue derivative molecule
  • the purpose of the solution is to fully contact the first stabilizer with the nuclide solution, quench a large number of free radicals caused by radiolysis in the nuclide solution, and thereby protect the Evans blue derivative molecules added to the reaction system from being activated. Free radical attack ensures the initial radiochemical purity of the product. This process can make the initial radiochemical purity reach 95.0-99.5%, while the initial radiochemical purity of the product obtained by the synthesis process of directly mixing the solution containing the Evans blue derivative molecules with the nuclide solution is about 89%-93 %.
  • the process method in this application can at least ensure that the labeling rate is above 90%, preferably above 95%, and optimally above 99%.
  • there is no purification step to remove radioactive impurities after the reaction such as preparative liquid phase separation, solid phase extraction separation, etc.
  • the step of recovering the product also includes passing the obtained solution through a 0.22 ⁇ m sterilizing filter membrane, and further diluting the solution according to the dosage.
  • the present application relates to an aqueous radiopharmaceutical solution, which includes a complex formed by a targeting molecule modified by an Evans blue (EB) fragment and a radioactive metal nuclide, and a stabilizer.
  • EB Evans blue
  • the targeting molecule modified with an Evans blue (EB) fragment is a compound represented by formula I or its Pharmaceutically acceptable esters, amides, solvates, salts, or salts of compounds represented by formula I or pharmaceutically acceptable esters thereof, or compounds represented by formula I or pharmaceutically acceptable amides thereof Salt, or the solvate of the compound represented by formula I or its pharmaceutically acceptable ester, or the solvate of the compound represented by formula I or its pharmaceutically acceptable amide, or the solvate represented by formula I A solvate of a compound or a pharmaceutically acceptable salt thereof.
  • EB Evans blue
  • L 1 is —(CH 2 ) m —, where m is an integer from 0 to 12, where each CH 2 may individually be replaced by —O—, —NH(CO)—, or —(CO)NH—, condition is that no two adjacent CH 2 groups are replaced;
  • L 3 is —(CH 2 ) n —, where n is an integer from 0 to 12, where each CH 2 may individually be replaced with —O—, —NH(CO)—, or —(CO)NH—, condition is that no two adjacent CH 2 groups are replaced;
  • Ch is a chelating group
  • Tg is the targeting group.
  • Evans Blue (EB) is a non-membrane permeable azo dye preparation. Because it has a high affinity with serum albumin in the blood, it uses its reversible binding property to albumin to intercept A short EB fragment (truncated Evans Blue, tEB) modifies the targeting molecule so that the targeting molecule can reversibly bind to endogenous serum albumin through the tEB fragment, and serum albumin is used as a reversible carrier of the drug molecule to prolong the drug
  • tEB truncated Evans Blue
  • the half-life of molecules in the blood increases the availability of drug molecules and further increases the accumulation and retention time of drug molecules in tumors.
  • the structural formula of Evans Blue (EB) dye is shown in Formula VI:
  • the Evans blue derivative molecule is a compound of formula I. In other specific embodiments, the Evans blue derivative molecule is a pharmaceutically acceptable ester, amide, solvate or salt of the compound of Formula I. In other specific embodiments, the Evans blue derivative molecule is a salt of a pharmaceutically acceptable ester of the compound of Formula I. In other specific embodiments, the Evans blue derivative molecule is a salt of a pharmaceutically acceptable amide of the compound of Formula I. In other specific embodiments, the Evans blue derivative molecule is a solvate of a pharmaceutically acceptable ester of the compound of Formula I. In other specific embodiments, the Evans blue derivative molecule is a solvate of a pharmaceutically acceptable amide of the compound of Formula I. In other specific embodiments, the Evans blue derivative molecule is a solvate of a pharmaceutically acceptable salt of the compound of Formula I.
  • the Evans blue derivative molecules can be synthesized by conventional chemical methods from parent compounds containing basic or acidic moieties.
  • L 1 in Formula I is -NH(CO)-
  • L 3 is -NH(CO)CH 2 -
  • the compound of Formula I is a compound of Formula VII:
  • the chelating group Ch in formula I is selected from Preferably, the chelating group Ch in formula I is
  • the chelating group has two or more coordinating atoms and can be combined with the same central atom to form a cyclic structure. It can form two or more separate groups with a single central atom, usually a metal ion. Coordination key.
  • the chelating group in this application is an organic group with multiple N, O or S heteroatoms, and has a structure that allows two or more heteroatoms to form bonds with the same metal ion. In a specific embodiment of the present application, the chelating group is used to form a bonded structure with a radioactive metal nuclide.
  • the targeting group Tg in formula I is a compound group that can specifically target a certain biological target.
  • the Tg is selected from the group consisting of: somatostatin receptor (SSTR), prostate specific membrane antigen (PSMA), fibroblast activation protein (FAP), folate receptor (FR), epidermal growth factor receptor or integrin compound group.
  • the targeting group Tg is selected from the group consisting of: somatostatin receptor (SSTR), prostate specific membrane antigen (PSMA), fibroblast activation protein (FAP), folate receptor (FR), epidermal growth factor receptor or integrin compound group.
  • the compound of formula I is EB-PSMA, and its structural formula is as shown in formula II:
  • the compound of formula I is DOTA-EB-TATE, and its structural formula is as shown in formula III:
  • the compound of formula I is EB-FAPI, and its structural formula is as shown in formula IV:
  • the compound of formula I is NMEB-RGD, and the structural formula is as shown in formula V:
  • the radioactive metal nuclide forming a complex with the Evans blue derivative molecule is selected from 177 Lu, 99m Tc, 68 Ga, 64 Cu, 67 Cu, 111 In, 86 Y, 90 Y , 89 Zr, 186 Re, 188 Re, 153 Sm, 82 Rb, 166 Ho, 225 Ac, 212 Pb, 213 Bi, 212 Bi, 227 Th.
  • the radioactive metal nuclide can be chelated or by other means, such as commonly known in the chemical field.
  • the regular covalent bond or ionic bond is combined with the chelating group Ch. Radionuclides may be suitable for purposes such as radiotherapy and/or diagnostics.
  • the radioactive metal nuclide is present in the pharmaceutical aqueous solution preparation at a volume radioactivity concentration of 0.037-1850 MBq/mL.
  • the stabilizer in the radiopharmaceutical aqueous solution is an anti-radioactive decomposition and degradation stabilizer.
  • the stabilizer is selected from the group consisting of gentisic acid and its salts, ascorbic acid and its salts, and the like. Amino acid, cysteine and its salts, methionine, selenomethionine, thiosulfate, maltose, inositol, benzyl alcohol, trehalose, povidone, nicotinamide, ethanol, curcumin,
  • One or more of melatonin is preferably one or more of gentisic acid, ethanol, and methionine.
  • the total concentration of the stabilizer in the aqueous drug solution is 0.5-400 mg/mL, for example, it can be 0.5, 1, 2, 5, 10, 25, 50, 100, 150, 200, 250 , 300, 350, 400mg/mL.
  • it is 1-80 mg/mL, for example, it can be 1, 2, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80 mg/mL .
  • the stabilizer is added during the complexation reaction to form the nuclide complex and after the reaction is completed.
  • adding during the complexation reaction means that the stabilizer, the radionuclide solution forming the complex and the Evans blue derivative molecule solution jointly form a reaction phase solution when conditions sufficient for the complexation reaction to occur are reached.
  • the adding after the reaction is completed means that the stabilizer is added after the complexing reaction has occurred for a certain period of time and the complex has been formed.
  • the stabilizer added during the complexation reaction is the first stabilizer, and the stabilizer added after the reaction is completed is the second stabilizer.
  • the first stabilizer is generally a small molecule compound with antioxidant properties to reduce radiolysis under high radiation.
  • the main function of the secondary stabilizer is to maintain the stability of the formulation during storage.
  • the first stabilizer and the second stabilizer may be selected from the same stabilizer or from different stabilizers.
  • the first stabilizer is selected from the group consisting of gentisic acid and its salts, ascorbic acid and its salts, histidine, cysteine and its salts, methionine, selenomethionine, sulfur One or more of sulfate, maltose, inositol, benzyl alcohol, trehalose, povidone, nicotinamide, ethanol, curcumin, and melatonin, preferably gentisic acid.
  • the second stabilizer is selected from the group consisting of gentisic acid and its salts, ascorbic acid and its salts, histidine, cysteine and its salts, methionine, selenomethionine, sulfur Sulfate, maltose, inositol, benzyl alcohol, trehalose, povidone, nicotinamide, ethanol, curcumin, melatonin
  • gentisic acid and its salts ascorbic acid and its salts, histidine, cysteine and its salts, methionine, selenomethionine, sulfur Sulfate, maltose, inositol, benzyl alcohol, trehalose, povidone, nicotinamide, ethanol, curcumin, melatonin
  • gentisic acid and its salts ascorbic acid and its salts
  • histidine cysteine and its salts
  • methionine methionine
  • the first stabilizer and the second stabilizer are the same and are selected from gentisic acid or its salt.
  • gentisic acid i.e., the first stabilizer
  • concentration range in the reaction system is 0.6-20 mg/mL, preferably 2-10 mg/mL, for example, it can be 2, 2.5 , 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10mg/mL.
  • gentisic acid i.e., the first stabilizer
  • concentration range in the reaction system is 0.6-20 mg/mL, preferably 2-10 mg/mL, for example, it can be 2, 2.5 , 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10mg/mL.
  • gentisic acid i.e.
  • the second stabilizer to the preparation after the reaction, so that gentisic acid exists in the entire drug aqueous solution at a total concentration of 0.1-10mg/mL, preferably 0.5-5mg/mL, for example It can be 0.5, 0.8, 1.0, 1.2, 1.5, 2.0, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 3.0, 3.2, 3.5, 3.8, 4.0, 4.5, 5.0mg/mL.
  • the stabilizers are two different stabilizers.
  • the first stabilizer added to the reaction system during the complexation reaction is gentisic acid or a salt thereof. It exists in the pharmaceutical aqueous solution at a concentration of 0.5-5 mg/mL, preferably 0.5-2 mg/mL, for example, it can be 0.5, 0.8, 1.0, 1.2, 1.5, 1.8, 2.0 mg/mL.
  • the second stabilizer added after the reaction is ethanol, which exists in the drug aqueous solution at a concentration of 0-400 mg/mL, preferably 10-120 mg/mL, for example, it can be 10, 30, 50, 60, 70, 80 ,100,120mg/mL.
  • the volume fraction is 0%-50%, preferably 1%-15%, for example, it can be 1%, 3%, 5%, 6%, 7%, 8%, 9%, 10%, 12%, 15% .
  • the first stabilizer added to the reaction system during the complexation reaction is gentisic acid or a salt thereof, which is present in the aqueous drug solution at a concentration of 0.5-5 mg/mL, preferably 0.5-2mg/mL, for example, it can be 0.5, 0.8, 1.0, 1.2, 1.3, 1.4, 1.5, 1.6, 1.8, 2.0mg/mL.
  • the second stabilizer added after the reaction is L-methionine, which exists in the drug aqueous solution at a concentration of 0-50 mg/mL, preferably 1-10 mg/mL, for example, it can be 1, 2, 3, 4, 5, 6, 7, 8, 9, 10mg/mL.
  • the two stabilizers preferably do not contain ascorbic acid and its salts.
  • the aqueous drug solution further includes a buffer.
  • the buffer can be added during the complexation reaction to adjust the pH of the reaction phase solution, or can be added again after the reaction to adjust the pH of the preparation solution.
  • the buffer added twice can be the same or different.
  • the buffer can be an acetate system (such as acetic acid-sodium acetate system, sodium acetate system), a citrate system (such as citric acid-sodium citrate system), or a phosphate system (such as sodium dihydrogen phosphate-disodium hydrogen phosphate system), formate system (such as formic acid-sodium formate system) solution.
  • the concentration of buffer salt in the reaction phase solution is 0.01-2.0M.
  • the total buffer salt concentration in the final aqueous drug solution is 0.005-0.5M.
  • the aqueous drug solution also includes a co-solvent, which functions to reduce the adsorption of API on various contact material surfaces (especially glass and plastic surfaces).
  • the co-solvent is selected from the group consisting of polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 80, poloxamer 188, polyoxyethylene castor oil, Span, ethanol, propylene glycol, glycerin, polyoxyethylene One or more of ethylene glycol (average molecular weight: 200 to 8000), sorbitol, dimethyl sulfoxide, and sodium lauryl sulfate, preferably polysorbate 80.
  • the concentration of the co-solvent in the aqueous drug solution is 0.01-10 mg/mL, preferably 0.05-1.0 mg/mL, for example, it can be 0.05, 0.08, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 1.0mg/mL.
  • the aqueous pharmaceutical solution further includes a chelating agent for free metal nuclides.
  • the function of the chelating agent is to complex with unreacted free nuclide ions in the drug aqueous solution to reduce unnecessary irradiation of healthy tissues by free radionuclide ions in the body. Therefore, the chelating agent is required to have a strong ability to complex with nuclide ions. Even after the injection enters the organism and is diluted by plasma, it can still react quickly with free nuclide ions at a lower concentration. The complexation reaction needs to be rapid, the conditions are mild, and it can be completely carried out at room temperature.
  • the chelating agent is pentetic acid or a salt thereof, preferably pentetic acid.
  • the concentration of the chelating agent in the aqueous drug solution is 0.005-0.1 mg/mL, for example, it can be 0.005, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1 mg/mL.
  • pentetic acid has sufficient complexing ability for free nuclide ions, and the complex can maintain stability for at least 48 hours under radiation decomposition, and preferably can maintain stability for 72 hours, that is There will be no release of free nuclide ions due to radiolysis of the chelating agent.
  • This application also provides a solution for applying the pharmaceutical aqueous solution in radiotherapy and/or diagnosis of tumors, including by administering an effective amount of the pharmaceutical aqueous solution to the patient, or combining it with one or more other tumor immunology therapeutic agents.
  • a common composition can be used to treat neuroendocrine tumors, prostate cancer, breast cancer, ovarian cancer, pancreatic cancer, liver cancer, lung cancer, colorectal cancer, melanoma, etc.
  • the pharmaceutical aqueous solution provided in this application or the composition containing the same can also be used to prepare drugs for preventing or treating diabetes and Alzheimer's disease.
  • using the pharmaceutical aqueous solution provided in this application can at least reduce the radiochemical purity of the API within 48 hours under storage conditions of 32°C and 60% RH. Less than 90%, more preferably not less than 90% radiochemical purity within 72 hours, the radiochemical purity is the value measured by HPLC.
  • This application also relates to a method for preparing an aqueous radiopharmaceutical solution, which in a specific embodiment includes the following steps:
  • the reaction vessel After a given time, add the solution containing the Evans blue derivative molecules to the reaction vessel.
  • the given time is 0.1 to 20 minutes, and more preferably 3 to 10 minutes;
  • the Evans blue derivative molecule reacts with a radionuclide to obtain the radionuclide complex
  • the Evans blue derivative molecule is a compound represented by Formula I or a pharmaceutically acceptable ester, amide, solvate, salt thereof, or a compound represented by Formula I or a pharmaceutically acceptable ester thereof
  • the salt, or the salt of the compound represented by formula I or its pharmaceutically acceptable amide, or the solvate of the compound represented by formula I or its pharmaceutically acceptable ester, or the salt represented by formula I A solvate of a compound or a pharmaceutically acceptable amide thereof, or a solvate of a compound represented by formula I or a pharmaceutically acceptable salt thereof,
  • L 1 is —(CH 2 ) m —, where m is an integer from 0 to 12, where each CH 2 may individually be replaced by —O—, —NH(CO)—, or —(CO)NH—, condition is that no two adjacent CH 2 groups are replaced;
  • L 3 is —(CH 2 ) n —, where n is an integer from 0 to 12, where each CH 2 may individually be replaced with —O—, —NH(CO)—, or —(CO)NH—, condition is that no two adjacent CH 2 groups are replaced;
  • Ch is a chelating group
  • Tg is the targeting group.
  • the aqueous radiopharmaceutical solution contains a radionuclide complex formed by a radionuclide and the Evans blue derivative molecule.
  • the solution containing radionuclides is a solution containing radioactive metal elements.
  • the radionuclides are selected from 177 Lu, 99m Tc, 68 Ga, 64 Cu, 67 Cu, 111 In, 86 Y, 90 Y, 89 Zr, 186 Re, 188 Re, 153 Sm, 82 Rb, 166 Ho, 225 Ac, 212 Pb, 213 Bi, 212 Bi or 227 Th.
  • the radionuclide is 177 Lu, and in the step of complexing reaction with Evans blue derivative molecules, the specific activity of the radionuclide is not less than 20Ci/mg, It is preferably not less than 60Ci/mg, and most preferably not less than 80Ci/mg. Radionuclides with too low specific activity will affect radioactive labeling efficiency.
  • the Evans Blue derivative molecule is a targeting molecule modified with a truncated Evans Blue fragment (truncated Evans Blue, tEB).
  • the Evans blue derivative molecule is a compound of Formula I.
  • the Evans blue derivative molecule is a pharmaceutically acceptable ester, amide, solvate or salt of the compound of Formula I.
  • the Evans blue derivative molecule is a salt of a pharmaceutically acceptable ester of the compound of Formula I.
  • the Evans blue derivative molecule is a salt of a pharmaceutically acceptable amide of the compound of Formula I.
  • the Evans blue derivative molecule is a solvate of a pharmaceutically acceptable ester of the compound of Formula I.
  • the Evans blue derivative molecule is a solvate of a pharmaceutically acceptable amide of the compound of Formula I. In other specific embodiments, the Evans blue derivative molecule is a solvate of a pharmaceutically acceptable salt of the compound of Formula I.
  • L 1 in Formula I is -NH(CO)-
  • L 3 is -NH(CO)CH 2 -
  • the compound of Formula I is a compound of Formula VII:
  • the chelating group Ch in formula I is selected from Preferably, the chelating group Ch in formula I is
  • the targeting group Tg in formula I is a compound group that can specifically target a certain biological target.
  • the Tg is selected from the group consisting of capable of targeting somatostatin receptor (SSTR), prostate specific membrane antigen (PSMA), fibroblast activation protein (FAP), folate receptor (FR), epidermal growth factor receptor The compound group of body or integrin. exist
  • the targeting group Tg is selected from
  • the compound of formula I is EB-PSMA, and its structural formula is as shown in formula II:
  • the compound of formula I is DOTA-EB-TATE, and its structural formula is as shown in formula III:
  • the compound of formula I is EB-FAPI, and its structural formula is as shown in formula IV:
  • the compound of formula I is NMEB-RGD, and the structural formula is as shown in formula V:
  • the solution containing radionuclides is taken out from the raw material bottle and then added to the reaction vessel, and after the solution containing radionuclides is taken out, it is rinsed with a rinse solution Rinse the raw material bottle to extract the nuclide solution remaining in the raw material bottle, and transfer the rinsed solution into the reaction vessel to mix with the solution containing radioactive nuclide.
  • the flushing solution is an aqueous solution, preferably selected from a solution containing a first stabilizer, a solution containing buffer salt, water or sodium chloride injection.
  • the flushing solution is selected from water for injection or sodium chloride injection.
  • the flushing is repeated one or more times using the flushing liquid.
  • the first stabilizer is selected from the group consisting of gentisic acid and its salts, ascorbic acid and its salts, histidine, cysteine and its salts, methionine, selenium methionine One or two or more of amino acid, thiosulfate, maltose, inositol, benzyl alcohol, trehalose, povidone, nicotinamide, ethanol, curcumin, and melatonin, preferably gentisic acid.
  • a solution containing the first stabilizer and a solution containing a radionuclide are mixed in a reaction vessel, and after a given time, the solution containing the Evans blue derivative molecule is mixed Add to the reaction vessel.
  • the given time can make the first stabilizer fully contact with the nuclide solution, quench a large number of free radicals caused by radiolysis existing therein, thereby protecting the Evans blue derivative molecules subsequently added to the reaction system from active free radicals. Attack is beneficial to improving the initial radiochemical purity of the final product.
  • the given time is 0.1 to 20 minutes, for example, it can be 0.1, 0.2, 0.3, 0.5, 0.8, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5 , 5.5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 minutes; more preferably 3 to 10 minutes, for example, it can be 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10 minutes.
  • the solution containing Evans blue derivative molecules is added to the reaction phase solution to react with the radionuclide to obtain the radionuclide complex.
  • the solution of the Evans blue derivative molecule is selected from a compound solution with a concentration of 0.05-10.0 mg/mL, and the preparation method is to dissolve the lyophilized powder of the labeling precursor in Sterile water for injection or ethanol.
  • the radionuclide complex is 177 Lu-DOTA-EB-TATE.
  • the first stabilizer present is gentisic acid, which is mentioned above
  • concentration in the reaction phase is 0.6-20.0mg/mL, preferably 2-10mg/mL, for example, it can be 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10mg/mL.
  • the most preferred range is 3.0-5.0 mg/mL, for example, it can be 3.0, 3.2, 3.4, 3.5, 3.6, 3.8, 4.0, 4.2, 4.5, 4.8, 5.0 mg/mL.
  • the molar ratio between the Evans blue derivative molecule and the radionuclide is 1.5- 50, preferably 5-20, for example, it can be 5, 8, 10, 12, 15, 18, 20.
  • the molar ratio refers to the molar ratio of the Evans blue derivative molecules (labeling precursor) to the radioactive nuclide in the reaction system.
  • the increase in molar ratio is conducive to the complete reaction of radionuclides and increases the labeling rate.
  • unlabeled labeling precursors will compete with the API in vivo.
  • an excessively low molar ratio causes the API to lack a carrier and is easily bound to and lost by other non-specific targets in the body, failing to achieve the expected therapeutic or diagnostic effect.
  • the concentration of the reaction phase in the reaction phase solution can also be controlled. Theoretically, the higher the concentration of the reaction phase, the faster the labeling reaction rate, but at the same time the stronger the radiation decomposition effect caused by radionuclides, so the concentration of the reaction phase cannot be too high, and the concentration of the reaction phase that is too low will cause the reaction volume to become Large, limiting the production of large quantities of nuclide complexes.
  • the molecular concentration of the Evans blue derivative is in the range of 0.01-1.0mg/mL, preferably 0.05-0.5mg/mL, for example, it can be 0.05, 0.1, 0.15, 0.2, 0.25, 0.3, 0.35, 0.4, 0.45, 0.5mg/mL.
  • the reaction temperature and time are controlled to achieve a reaction labeling rate of >90% and chemical purity. >90%, radiochemical purity >90%.
  • the reaction temperature is 50-100°C, preferably 60-80°C, for example, it can be 60, 62, 65, 68, 70, 72, 75, 78, 80°C;
  • the reaction time is 5-60 minutes, for example, it can be 5, 10, 12, 15, 18, 20, 25, 30, 40, 50, 60 minutes, preferably 10-30 minutes, most preferably 10-20 minutes, for example, it can be 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 minutes.
  • the second stabilizer is added.
  • the second stabilizer is selected from the group consisting of gentisic acid and its salts, ascorbic acid and its salts, histidine, cysteine and its salts, methionine, selenomethionine, and thiosulfate. , one or more of maltose, inositol, benzyl alcohol, trehalose, povidone, nicotinamide, ethanol, curcumin, and melatonin, preferably gentisic acid, ethanol, or methionine.
  • the concentration of the second stabilizer is 0-400 mg/mL, for example, it can be 0, 0.5, 1, 2, 5, 10, 25, 50, 100, 150, 200, 250, 300, 350, 400mg/mL.
  • the preparation method further includes adding a buffer salt solution before the Evans blue derivative molecule reacts with the radionuclide.
  • the buffer salt solution is present in the first solution containing stabilizer solution.
  • the buffered salt solution is selected from acetate, citrate, phosphate or formate solutions, preferably acetic acid-sodium acetate buffered salt solution.
  • buffer salt solution can adjust the pH value of the reaction system, and control the pH value of the reaction phase system in the range of 3.5-6.0, for example, it can be 3.5, 3.8, 4, 4.2, 4.4, 4.6, 4.8, 5, 5.5, 6 , the preferred pH value is 3.5-5.
  • the pH value of the final formulation solution is controlled to be 4-6, for example, it can be 4, 4.2, 4.5, 4.8, 5, 5.2, 5.5, 5.8, 6.
  • the step of adding the solution containing the second stabilizer to the reaction vessel after a given reaction time further includes adding a co-solvent to the reaction vessel.
  • the co-solvent is selected from polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 80, poloxamer 188, polyoxyethylene castor oil, Span, ethanol , one or more of propylene glycol, glycerol, polyethylene glycol (average molecular weight 200-8000), sorbitol, dimethyl sulfoxide, sodium lauryl sulfate, preferably polysorbate 80 .
  • the co-solvent is added so that its concentration in the aqueous drug solution is 0.01-10 mg/mL, preferably 0.05-1.0 mg/mL, for example, it can be 0.05, 0.08, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 1.0mg/mL.
  • the step of adding the solution containing the second stabilizer to the reaction vessel after a given reaction time also includes adding a free nuclide chelating agent to the reaction vessel, and the chelating agent can Selected from pentetic acid and its salts, preferably pentetic acid.
  • the chelating agent is added so that its concentration in the aqueous drug solution is 0.005-0.1 mg/mL, for example, it can be 0.005, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1mg/mL.
  • the preparation method of the present application also includes filtering and sterilizing the radiopharmaceutical aqueous solution.
  • the radiopharmaceutical aqueous solution is filtered and sterilized through a 0.22 ⁇ m filter membrane.
  • the preparation method of the present application also includes diluting the radioactive aqueous solution, preferably by adding sodium chloride injection for dilution for recovery.
  • the filtration sterilization and dilution are performed after adding the solution containing the second stabilizer.
  • This application does not limit the order of the filtration sterilization and dilution steps. That is, it can be filtered and sterilized first and then diluted, or it can be diluted first, then filtered and sterilized through a filter membrane, and then recovered.
  • the present application provides a method for preparing 177 Lu-DOTA-EB-TATE radiopharmaceutical aqueous solution in the following sequence:
  • the precursor EB-PSMA used in the following examples was synthesized according to the literature method (Bioconjugate Chem. 2018, 29, 3213-3221).
  • the precursor DOTA-EB-TATE used in the following examples was synthesized according to the literature method (Theranostics. 2018; 8:735-745).
  • the precursor EB-FAPI used in the following examples was synthesized according to the literature method (Theranostics. 2022; 12(1): 422-433).
  • the gentisic acid used in the following examples was purchased from Chengdu Prefa Technology Development Co., Ltd., and the pentetic acid was purchased from Jiangxi Alpha Hi-Tech Pharmaceutical Co., Ltd.
  • Example 1 Selection of stabilizers in aqueous pharmaceutical solutions
  • reaction phase solution Preparation of reaction phase solution: Add 10mCi carrier-free lutetium chloride [ 177 Lu] solution (approximately 10 ⁇ L), 20 ⁇ L formic acid-sodium formate buffer salt solution (containing 50mg/mL gentisic acid), and 60 ⁇ L water for injection into the reaction vessel, and mix After homogenization, let the mixed solution stand at room temperature for 3 minutes. Then continue to add 10 ⁇ L of DOTA-EB-TATE precursor solution into the reaction vessel and mix evenly. The mixed solution is the reaction phase solution.
  • Heating reaction and cooling Place the above reaction phase solution in a heater preheated to 90°C to react for 15 minutes. After the reaction is completed, take out the reaction vessel and cool it for 15 minutes.
  • auxiliary solution contains 45mg/mL gentisic acid, 0.5mg/mL pentic acid and 2.0mg/mL polysorbate. Ester 80. Finally, sodium chloride injection was added to the reaction vessel to dilute the total volume to 1.0 mL to obtain the final preparation solution.
  • the final formulation solution contains 10 mg/mL gentisic acid, 0.1 mg/mL pentetic acid, and 0.4 mg/mL polysorbate 80.
  • the activity concentration of the API molecule [ 177 Lu]Lu-DOTA-EB-TATE at the calibration time is is 10mCi/mL, and the calibration time refers to the production end time (T 0 ).
  • the formulation solution was stored in a stability box, the storage temperature was set to 32°C, and the storage humidity was set to 60% RH.
  • the radiochemical purity of the preparation solution was detected by Radio-HPLC at time T 0 and was 100%, and the radiochemical purity of the preparation solution was detected by ITLC at time T 0 and was 100%.
  • reaction phase solution Preparation of the reaction phase solution: Add 10mCi carrier-free lutetium chloride [ 177 Lu] solution (about 10 ⁇ L), 20 ⁇ L formic acid-sodium formate buffer salt solution (containing 50mg/mL gentisic acid), and 160 ⁇ L water for injection into the reaction vessel, and mix After homogenization, let the mixed solution stand at room temperature for 3 minutes. Then continue to add 10 ⁇ L of DOTA-EB-TATE precursor solution into the reaction vessel and mix evenly. The mixed solution is the reaction phase solution.
  • Heating reaction and cooling Place the above reaction phase solution in a heater preheated to 65°C to react for 40 minutes. After the reaction is completed, take out the reaction vessel and cool it for 15 minutes.
  • Preparation and dilution After the reaction phase solution cools to room temperature, add 100 ⁇ L to the reaction container.
  • the auxiliary solution contains 0.3 mg/mL pentetic acid and 1.0 mg/mL polysorbate 80. Then add 50 mg of absolute ethanol, and finally add sodium chloride injection to the reaction vessel to dilute the total volume to 1.0 mL to obtain the final preparation solution.
  • the final formulation solution contains 1mg/mL gentisic acid, 50mg/mL ethanol, 0.03mg/mL pentetic acid, 0.1mg/mL polysorbate 80, in which the API molecule [ 177 Lu]Lu-DOTA-EB-TATE is calibrated
  • the activity concentration at the time is 10mCi/mL, and the calibration time refers to the production end time (T 0 ).
  • the formulation solution was stored in a stability box, the storage temperature was set to 32°C, and the storage humidity was set to 60% RH.
  • the radiochemical purity of the preparation solution was detected by Radio-HPLC at time T 0 and was 100%, and the radiochemical purity of the preparation solution was detected by ITLC at time T 0 and was 100%.
  • reaction phase solution Preparation of reaction phase solution: Add 20mCi carrier-free lutetium chloride [ 177 Lu] solution (about 20 ⁇ L), 20 ⁇ L ammonium acetate buffer salt solution (containing 50mg/mL gentisic acid), and 40 ⁇ L water for injection into the reaction vessel, and mix evenly Then, let the mixed solution stand at room temperature for 10 minutes. Then continue to add 20 ⁇ L of EB-FAPI precursor solution into the reaction vessel and mix evenly. The mixed solution is the reaction phase solution.
  • Heating reaction and cooling Place the above reaction phase solution in a heater preheated to 95°C to react for 30 minutes. After the reaction is completed, take out the reaction vessel and cool it for 15 minutes.
  • auxiliary solution contains 20mg/mL methionine, 4.0mg/mL gentisic acid, 0.2mg/mL spray acid and 0.8mg/mL polysorbate 80.
  • sodium chloride injection was added to the reaction vessel to dilute the total volume to 1.0 mL to obtain the final preparation solution.
  • the final preparation solution contains 3.0mg/mL gentisic acid, 10mg/mL methionine, 0.1mg/mL pentetic acid, 0.4mg/mL polysorbate 80, in which the API molecule [ 177 Lu]Lu-EB-FAPI
  • the activity concentration at the calibration time is 20mCi/mL, and the calibration time refers to the production end time (T 0 ).
  • the formulation solution was stored in a stability box, the storage temperature was set to 32°C, and the storage humidity was set to 60% RH.
  • the radiochemical purity of the preparation solution was detected by Radio-HPLC at T 0 time and was 98%, and the radiochemical purity of the preparation solution was detected by ITLC at T 0 time and was 100%.
  • reaction phase solution Preparation of the reaction phase solution: Add 10mCi carrier-free lutetium chloride [ 177 Lu] solution (about 10 ⁇ L), 20 ⁇ L acetic acid-sodium acetate buffered salt solution (containing 50mg/mL gentisic acid), and 60 ⁇ L water for injection into the reaction vessel. After mixing evenly, let the mixed solution stand at room temperature for 3 minutes. Then continue to add 10 ⁇ L of EB-PSMA precursor solution into the reaction vessel and mix evenly. The mixed solution is the reaction phase solution.
  • Heating reaction and cooling Place the above reaction phase solution in a heater preheated to 80°C to react for 15 minutes. After the reaction is completed, take out the reaction vessel and cool it for 15 minutes.
  • auxiliary solution contains 30mg/mL ascorbic acid, 1.0mg/mL pentetic acid and 4.0mg/mL polysorbate 80 .
  • sodium chloride injection was added to the reaction vessel to dilute the total volume to 1.0 mL to obtain the final preparation solution.
  • the final formulation solution contains 1.0mg/mL gentisic acid, 3.0mg/mL ascorbic acid, 0.1mg/mL pentetic acid, 0.4mg/mL polysorbate 80, in which the API molecule [ 177 Lu]Lu-EB-PSMA is calibrated
  • the activity concentration at the time is 10mCi/mL, and the calibration time refers to the production end time (T 0 ).
  • the formulation solution was stored in a stability box, the storage temperature was set to 32°C, and the storage humidity was set to 60% RH.
  • the radiochemical purity of the preparation solution was detected using Radio-HPLC at T 0 time and was 93%.
  • the aqueous drug solution prepared by using gentisic acid, gentisic acid and ethanol, gentisic acid and methionine as stabilizers in the above prescriptions (1) to (3) can be obtained in both 48h and 72h. More than 90% of the API is radiochemically pure, which is significantly better than the ascorbic acid stabilizer selection used in the prescription (4). This shows that the pharmaceutical aqueous solution provided by the present invention can maintain better stability.
  • Heating reaction and cooling Place the above reaction phase solution at room temperature or in a heater preheated to different temperatures for 120 minutes.
  • the temperatures examined include room temperature, 50°C, 60°C, 70°C, 80°C, 90°C, 95°C, and 100°C.
  • reaction temperature and reaction time are two complementary process conditions. “React at a lower temperature for a longer time” or “React at a higher temperature for a shorter time” can both increase the reaction labeling rate (ITLC ) ⁇ 99%, at this point we consider that the labeling reaction has completely progressed.
  • ITLC reaction labeling rate
  • Preparation of the reaction phase solution Add 10mCi carrier-free lutetium chloride [ 177 Lu] solution (about 10 ⁇ L) and 20 ⁇ L formic acid-sodium formate buffer salt solution (containing 50mg/mL gentisic acid) into the reaction vessel, and add this mixed solution in Let sit at room temperature for 3 minutes. Then continue to add different volumes of water for injection and DOTA-EB-TATE precursor solution to the reaction vessel, so that the molar ratios of DOTA-EB-TATE and lutetium chloride [ 177 Lu] in the reaction phase are 1, 1.5, and 1.5 respectively. 2, 3, 5, 10, 15, 30, 50, and make the total volume of the solution 0.1 mL. Mix the solution evenly. The mixed solution is the reaction phase solution.
  • Heating reaction and cooling Same as prescription (1).
  • the labeling rate is ⁇ 95%.
  • the labeling rate is ⁇ 99%. At this time, we think The labeling reaction is complete.
  • reaction phase solution A Preparation of reaction phase solution A: Add 10 mCi carrier-free lutetium chloride [ 177 Lu] solution (about 10 ⁇ L), 10 ⁇ L EB-FAPI precursor solution, 60 ⁇ L water for injection, and 20 ⁇ L acetic acid-sodium acetate buffered saline solution (approximately 10 ⁇ L) to the reaction vessel. It contains 50 mg/mL gentisic acid), mix it evenly, and the mixed solution is reaction phase solution A.
  • reaction phase solution B Preparation of reaction phase solution B: Add 10mCi carrier-free lutetium chloride [ 177 Lu] solution, 20 ⁇ L acetic acid-sodium acetate buffered salt solution (containing 50mg/mL gentisic acid), and 60 ⁇ L water for injection into the reaction vessel, and mix evenly. , let the mixed solution stand at room temperature for 3 minutes (ie, quenching time). Then continue to add 10 ⁇ L of EB-FAPI precursor solution into the reaction vessel and mix evenly.
  • the mixed solution is reaction phase solution B.
  • reaction phase solution C The rest is the same as reaction phase solution B except that the quenching time is 15 minutes.
  • Heating reaction and cooling same as prescription (1).
  • the radiochemical purity of reaction phase solutions A, B, and C are 93%, 99%, and 99% respectively.
  • Experimental results show that before adding the precursor solution, the mixed solution containing the nuclide solution, buffer salt and first stabilizer is allowed to stand for a short period of time (quenching time) before adding the precursor molecules for reaction.
  • the feeding sequence can Significantly improves the initial radiochemical purity of API. Because after a period of quenching time, the first stabilizer is in full contact with the nuclide solution, so that a large number of free radicals generated due to high radioactivity in the solution are quenched by the first stabilizer, thereby reducing the impact of free radicals when the precursor molecules are subsequently added. Mark the destruction of precursor molecules.
  • the quenching time depends on the type and initial activity of the nuclide. Generally speaking, the quenching time is controlled at 0.1-20 minutes, preferably 3-10 minutes.
  • Preparation of the reaction phase solution Elute the commercially available germanium and gallium generator with 0.1M hydrochloric acid to obtain a 68 Ga hydrochloric acid solution. Take 5 mCi of the 68 Ga hydrochloric acid solution in the reaction vessel and add 20 ⁇ L of sodium acetate solution (which contains 50 mg/mL gentisic acid). , 150 ⁇ L water for injection, mix evenly, let the mixed solution stand at room temperature for 6 minutes, then add 5 ⁇ L EB-FAPI precursor solution to the reaction vessel, Add water for injection to bring the total volume of the solution to 0.3 mL, and mix evenly. The mixed solution is the reaction phase solution.
  • Heating reaction and cooling Place the above reaction phase solution in a heater preheated to 95°C to react for 30 minutes. After the reaction is completed, take out the reaction vessel and cool it for 5 minutes.
  • auxiliary solution contains 45mg/mL gentisic acid, 0.5mg/mL pentetic acid and 2.0mg/mL polysorbate 80. Finally, add sodium chloride injection to the product bottle to dilute the total volume to 1.0 mL to obtain the final formulation solution.
  • the final formulation solution contains 9 mg/mL gentisic acid, 40% ethanol (i.e. 315.6 mg/mL), 0.1 mg/mL pentetic acid, and 0.4 mg/mL polysorbate 80, in which the API molecule [ 177 Lu ]
  • the activity concentration of Lu-EB-FAPI at the calibration time is 5mCi/mL, and the calibration time refers to the production end time (T 0 ).
  • the formulation solution was stored in a stability box, the storage temperature was set to 25°C, and the storage humidity was set to 60% RH.
  • the radiochemical purity of the preparation solution was detected by Radio-HPLC at T 0 time and was 99%, and the radiochemical purity of the preparation solution was detected by ITLC at T 0 time and was 100%.

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Abstract

本申请涉及放射性药物水溶液及其用途，所述放射性药物水溶液包括经伊文思蓝(EB)片段修饰的靶向分子与放射性金属核素形成的络合物以及稳定剂，所述稳定剂优选为龙胆酸、乙醇、甲硫氨酸中的一种或两种以上。

Description

放射性伊文思蓝衍生物药物水溶液及其制备方法和用途 技术领域
本申请涉及具有高化学稳定性和高放射化学稳定性的放射性药物水溶液及其制备方法，尤其涉及一种经伊文思蓝(EB)片段修饰的放射性核素络合物。
背景技术
药物分子在体内的循环半衰期能够很大程度上影响药物的有效性与可用性，尤其对于放射性标记的诊断和/或治疗类药物而言，放射性药物需要足够长的体内半衰期，以使得放射性药物与靶点充分结合，同时从其他非靶组织或器官逐渐清除，以实现其预期的诊断和/或治疗效果。在延长小分子药物的体内半衰期的方法中，对小分子进行结构修饰，使之在体内与人血清白蛋白(HSA)可逆结合，是一种常用的方法。伊文思蓝是一种偶氮染料，由于它对血清白蛋白具有高度亲和力，通常被用于血脑屏障完整性、血管通透性、血液体积和细胞活性的检测。使用伊文思蓝染料分子的结构片段对靶向分子(如DOTATATE、PSMA-617等)进行结构修饰，所得伊文思蓝衍生物分子，如DOTA-EB-TATE、EB-PSMA等，可通过分子与内源性血清白蛋白进行可逆的结合，将血清白蛋白作为药物分子的可逆载体，以延长药物分子在血液中的半衰期。已有文献(Bioconjugate Chem.2018,29,3213-3221)报道，伊文思蓝衍生物具有更长的体内循环半衰期、更高的肿瘤摄取率和肿瘤内更长的保留时间，并且由于其体内半衰期的延长，还可以进一步提高药物疗效、降低给药剂量与频次、减少药物毒性。
伊文思蓝染料的结构式如式Ⅵ所示：
在药物分子结构中引入的伊文思蓝片段与连接基团(Linker)的相对分子量较大，会极大地影响分子的理化性质，使得包含伊文思蓝衍生物的药物水溶液在处方工艺研究中面对更多挑战。例如获批用于治疗PSMA阳性的转移性去势抵抗性前列腺癌(mCRPC)药物PLUVICTOTM(lutetium Lu 177 vipivotide tetraxetan)注射液中,药物活性成分(API)为[177Lu]Lu-PSMA-617，稳定剂为0.39mg/mL龙胆酸、50.0mg/mL抗坏血酸钠。在用伊文思蓝片段对[177Lu]Lu-PSMA-617进行结构修饰后，所得分子[177Lu]Lu-EB-PSMA的稳定性质与[177Lu]Lu-PSMA-617完全不同，抗坏血酸及其盐的加入不仅不能发挥稳定剂的作用，反而还会加速[177Lu]Lu-EB-PSMA的辐射分解。
目前还没有用于肿瘤的放射治疗和/或诊断的包含伊文思蓝衍生物的放射性药物获批上市。因此，需要发展包含伊文思蓝衍生物的药物水溶液的稳定处方和适宜的生产工艺。
发明内容
本申请的目的是提供一种放射性药物水溶液，其包含经伊文思蓝(EB)片段修饰的靶向分子与放射性金属核素形成的络合物以及稳定剂。进一步的，本申请还提供所述放射性药物水溶液的制备方法。具体来说，本发明涉及以下内容：
1.放射性药物水溶液，其包括：如式Ⅰ所示的化合物或其药学上可接受的酯、酰胺、溶剂化物、盐、或如式Ⅰ所示的化合物或其药学上可接受的酯的盐、或如式Ⅰ所示的化合物或其药学上可接受的酰胺的盐、或如式Ⅰ所示的化合物或其药学上可接受的酯的溶剂化物、或如式Ⅰ所示的化合物或其药学上可接受的酰胺的溶剂化物、或如式Ⅰ所示的化合物或其药学上可接受的盐的溶剂化物，与放射性金属核素形成的络合物，以及以总浓度为0.5-400mg/mL存在的稳定剂，
其中，
L1是—(CH2)m—，其中m是0至12的整数，其中每个CH2可以单独地用—O—、—NH(CO)—、或—(CO)NH—替换，条件是没有两个相邻的CH2基团被替换；
L2是C1-C60连接基团，任选地包括—O—、—S—、—S(O)—、—S(O)2—、—N(R)—、—C(＝O)—、—C(＝O)O—、—OC(＝O)—、—N(R)C(＝O)—、—C(＝O)N(R)—、—OC(＝O)O—、—N(R)C(＝O)O—、—OC(＝O)N(R)—、其中每个R为H或C1-C6烷基；
L3是—(CH2)n—，其中n是0至12的整数，其中每个CH2可以单独地用—O—、—NH(CO)—、或—(CO)NH—替换，条件是没有两个相邻的CH2基团被替换；
Ch是螯合基团；
Tg是靶向基团。
2.根据项1所述的药物水溶液，其中，L1为—NH(CO)—，L3为—NH(CO)CH2—。
3.根据项1所述的药物水溶液，其中，Ch选自 优选为
4.根据项1所述的药物水溶液，其中，Tg选自能够靶向生长抑素受体(SSTR)、前列腺特异性膜抗原(PSMA)、成纤维细胞活化蛋白(FAP)、叶酸受体(FR)、表皮生长因子受体或整合素的化合基团。
5.根据项4所述的药物水溶液，其中，Tg选自
6.根据项1所述的药物水溶液，其中，式Ⅰ为式Ⅱ、式Ⅲ、式Ⅳ或式Ⅴ，

7.根据项1所述的药物水溶液，其中，所述放射性金属核素选自177Lu、99mTc、68Ga、64Cu、67Cu、111In、86Y、90Y、89Zr、186Re、188Re、153Sm、82Rb、166Ho、225Ac、212Pb、213Bi、212Bi或227Th。
8.根据项1～7中任一项所述的药物水溶液，其中，所述稳定剂选自龙胆酸及其盐、抗坏血酸及其盐、组氨酸、半胱氨酸及其盐、甲硫氨酸、硒甲硫氨酸、硫代硫酸盐、麦芽糖、肌醇、苯甲醇、海藻糖、聚维酮、烟酰胺、乙醇、姜黄素、褪黑素中的一种或两种以上，优选为龙胆酸、乙醇、甲硫氨酸中的一种或两种以上。
9.根据项8所述的药物水溶液，其中，在形成所述络合物的反应期间以及反应结束后分别加入所述稳定剂。
10.根据项9所述的药物水溶液，其中，在形成所述络合物的反应期间加入的稳定剂选自龙胆酸及其盐、抗坏血酸及其盐、组氨酸、半胱氨酸及 其盐、甲硫氨酸、硒甲硫氨酸、硫代硫酸盐、麦芽糖、肌醇、苯甲醇、海藻糖、聚维酮、烟酰胺、乙醇、姜黄素、褪黑素中的一种或两种以上，优选为龙胆酸；
在形成所述络合物的反应结束后加入的稳定剂选自龙胆酸及其盐、抗坏血酸及其盐、组氨酸、半胱氨酸及其盐、甲硫氨酸、硒甲硫氨酸、硫代硫酸盐、麦芽糖、肌醇、苯甲醇、海藻糖、聚维酮、烟酰胺、乙醇、姜黄素、褪黑素中的一种或两种以上，优选为龙胆酸、乙醇或甲硫氨酸。
11.根据项10所述的药物水溶液，其中，在形成所述络合物的反应期间和反应结束后加入的稳定剂均选自龙胆酸，且所述龙胆酸在所述药物水溶液中以0.1-10mg/mL的总浓度存在，优选为0.5-5mg/mL。
12.根据项10所述的药物水溶液，其中，在形成所述络合物的反应期间加入的稳定剂为龙胆酸，在形成所述络合物的反应结束后加入的稳定剂为乙醇，并且在所述药物水溶液中龙胆酸以0.1-10mg/mL的浓度存在，优选为0.5-5mg/mL，乙醇以0-400mg/mL的浓度存在。
13.根据项10所述的药物水溶液，其中，在形成所述络合物的反应期间加入的稳定剂为龙胆酸，在形成所述络合物的反应结束后加入的稳定剂为甲硫氨酸，并且在所述药物水溶液中龙胆酸以0.1-10mg/mL的浓度存在，优选为0.5-5mg/mL，甲硫氨酸以0-50mg/mL的浓度存在。
14.根据项10的药物水溶液，其中还包括缓冲液，所述缓冲液可选自醋酸盐、柠檬酸盐、磷酸盐或甲酸盐溶液；所述缓冲液中的缓冲盐在所述药物水溶液中的浓度为0.005-0.5M。
15.根据项10所述的药物水溶液，其中还包括助溶剂，所述助溶剂选自聚山梨酯20、聚山梨酯40、聚山梨酯60、聚山梨酯80、泊洛沙姆188、聚氧乙烯蓖麻油、司盘、乙醇、丙二醇、丙三醇、聚乙二醇(平均分子量为200～8000)、山梨醇、二甲基亚砜、十二烷基硫酸钠中的一种或两种以上，优选为聚山梨酯80。
16.根据项10所述的药物水溶液，其中还包括游离核素螯合剂，所述游离核素螯合剂可选自喷替酸及其盐；所述游离核素螯合剂在所述药物水溶液中以0.005-0.1mg/mL的浓度存在。
17.根据项1～16中任一项所述的药物水溶液，其特征在于，所述络合物在药物水溶液中以0.037-1850MBq/mL的活度浓度存在。
18.项1～17中任一项所述的药物水溶液在肿瘤的放射治疗和/或诊断中的用途。
19.制备放射性药物水溶液的方法，所述放射性药物水溶液包含放射性核素与伊文思蓝衍生物分子形成的络合物，其特征在于，包括如下步骤：
将含有第一稳定剂的溶液与含有放射性核素的溶液在反应容器中混合；
给定时间后，将含有所述伊文思蓝衍生物分子的溶液加入至所述反应容器，优选所述给定时间为0.1分钟～20分钟，进一步优选3分钟～10分钟；
所述伊文思蓝衍生物分子与放射性核素发生反应得到所述放射性核素络合物；
反应给定时间后向所述反应容器中加入含有第二稳定剂的溶液；
回收得到的所述放射性药物水溶液；
其中，所述伊文思蓝衍生物分子为如式Ⅰ所示的化合物或其药学上可接受的酯、酰胺、溶剂化物、盐、或如式Ⅰ所示的化合物或其药学上可接受的酯的盐、或如式Ⅰ所示的化合物或其药学上可接受的酰胺的盐、或如式Ⅰ所示的化合物或其药学上可接受的酯的溶剂化物、或如式Ⅰ所示的化合物或其药学上可接受的酰胺的溶剂化物、或如式Ⅰ所示的化合物或其药学上可接受的盐的溶剂化物，
其中，
L1是—(CH2)m—，其中m是0至12的整数，其中每个CH2可以单独地用—O—、—NH(CO)—、或—(CO)NH—替换，条件是没有两个相邻的CH2基团被替换；
L2是C1-C60连接基团，任选地包括—O—、—S—、—S(O)—、—S(O)2—、—N(R)—、—C(＝O)—、—C(＝O)O—、—OC(＝O)—、—N(R)C(＝O)—、—C(＝O)N(R)—、—OC(＝O)O—、—N(R)C(＝O)O—、—OC(＝O)N(R)—、 其中每个R为H或C1-C6烷基；
L3是—(CH2)n—，其中n是0至12的整数，其中每个CH2可以单独地用—O—、—NH(CO)—、或—(CO)NH—替换，条件是没有两个相邻的CH2基团被替换；
Ch是螯合基团；
Tg是靶向基团。
20.根据项19所述的方法，其特征在于，所述放射性核素选自177Lu、99mTc、68Ga、64Cu、67Cu、111In、86Y、90Y、89Zr、186Re、188Re、153Sm、82Rb、166Ho、225Ac、212Pb、213Bi、212Bi或227Th。
21.根据项19所述的方法，其特征在于，式Ⅰ中的Ch选自 优选为
22.根据项19所述的方法，其特征在于，式Ⅰ中的Tg选自能够靶向生长抑素受体(SSTR)、前列腺特异性膜抗原(PSMA)、成纤维细胞活化蛋白(FAP)、叶酸受体(FR)、表皮生长因子受体或整合素的化合基团。
23.根据项19所述的方法，其特征在于，所述伊文思蓝衍生物分子选自选自如式Ⅱ、式Ⅲ、式Ⅳ或式Ⅴ所示的化合物，
24.根据项19～23中任一项所述的方法，其中，所述含有放射性核素的溶液是从原料瓶中取出后加入至所述反应容器中，所述方法还包括：
以冲洗液对所述原料瓶进行冲洗，并将冲洗后的溶液转入所述反应容器中与所述含有放射性核素的溶液混合。
25.根据项24所述的方法，其特征在于，所述冲洗液为水溶液，优选地选自含有第一稳定剂的溶液、含有缓冲盐的溶液、水或氯化钠注射液；更优选地，使用冲洗液重复冲洗一次或更多次。
26.根据项19所述的方法，其特征在于，在所述伊文思蓝衍生物分子与放射性核素发生反应的步骤中，所述伊文思蓝衍生物分子与放射性核素之间的摩尔比为1.5–50，优选为5-20。
27.根据项19所述的方法，其特征在于，在所述伊文思蓝衍生物分子与放射性核素发生反应的步骤中，反应温度为50-100℃，优选为60-80℃，反应时间为5-60分钟，优选为10-30分钟。
28.根据项19所述的方法，其特征在于，所述第一稳定剂选自龙胆酸及其盐、抗坏血酸及其盐、组氨酸、半胱氨酸及其盐、甲硫氨酸、硒甲硫氨酸、硫代硫酸盐、麦芽糖、肌醇、苯甲醇、海藻糖、聚维酮、烟酰胺、乙醇、姜黄素、褪黑素中的一种或两种以上，优选为龙胆酸。
29.根据项28所述的方法，其特征在于，在所述伊文思蓝衍生物分子与放射性核素发生反应的步骤中，在反应相溶液中第一稳定剂的浓度为0.6-20.0mg/mL。
30.根据项19所述的方法，其特征在于，所述第二稳定剂选自龙胆酸及其盐、抗坏血酸及其盐、组氨酸、半胱氨酸及其盐、甲硫氨酸、硒甲硫氨酸、硫代硫酸盐、麦芽糖、肌醇、苯甲醇、海藻糖、聚维酮、烟酰胺、乙醇、姜黄素、褪黑素中的一种或两种以上，优选为龙胆酸、乙醇或甲硫氨酸。
31.根据项30所述的方法，其特征在于，在所述放射性药物水溶液中，所述第二稳定剂的浓度为0-400mg/mL。
32.根据项19所述的方法，其特征在于，在所述伊文思蓝衍生物分子与放射性核素发生反应之前加入缓冲盐溶液，优选地，所述缓冲盐溶液存在于所述含有第一稳定剂的溶液中。
33.根据项32所述的方法，其特征在于，所述缓冲盐溶液选自醋酸盐、柠檬酸盐、磷酸盐或甲酸盐溶液，优选为醋酸-醋酸钠缓冲盐溶液。
34.根据项19所述的方法，其特征在于，在反应给定时间后向所述反应容器中加入含有第二稳定剂的溶液的步骤还包括向所述反应容器中加入助溶剂。
35.根据项34所述的方法，其特征在于，所述助溶剂选自聚山梨酯20、聚山梨酯40、聚山梨酯60、聚山梨酯80、泊洛沙姆188、聚氧乙烯蓖麻油、司盘、乙醇、丙二醇、丙三醇、聚乙二醇(平均分子量为200～8000)、山梨醇、二甲基亚砜、十二烷基硫酸钠中的一种或两种以上，优选为聚山梨酯80。
36.根据项19所述的方法，其特征在于，在反应给定时间后向所述反应容器中加入含有第二稳定剂的溶液的步骤还包括向所述反应容器中加入游离核素螯合剂，所述螯合剂可选自喷替酸及其盐，优选为喷替酸。
37.根据项19～36中任一项所述的方法，其特征在于，还包括使放射性药物水溶液通过0.22μm滤膜过滤除菌，优选地，于加入含有第二稳定剂的溶液后进行所述过滤除菌。
38.根据项19～37中任一项所述的方法，其特征在于，还包括对所述放射性水溶液进行稀释，优选地，于加入含有第二稳定剂的溶液后，加入氯化钠注射液进行稀释。
39.放射性药物水溶液，其是由项19～38中任一项所述的方法制备得到。
40.根据项39所述的药物水溶液，其为项1～17中任一项所述的放射性药物水溶液。
41.项19～38中任一项所述的方法在肿瘤的放射治疗和/或诊断中的用途。
42.一种肿瘤的放射性治疗方法，其包括向有需要的受试者施用项1～17中任一项所述的药物水溶液。
发明效果
采用本申请提供的放射性药物水溶液具有如下有益效果：
在一个实施方案中，稳定剂可以为单独一种，也可以选择两种或两种以上的组合方式，在一些实施方案中，优选仅使用龙胆酸或龙胆酸与乙醇，乙醇具有良好的抗辐射分解的作用；同时，稳定剂优选不使用抗坏血酸。抗坏血酸的加入不利于伊文思蓝衍生物的稳定。
在一个实施方案中，在反应结束后添加喷替酸作为螯合剂，用以螯合未反应的游离核素离子，以减少游离的放射性核素离子在体内对健康组织 产生不必要的照射。
在一个实施方案中，采用本申请方案的药物水溶液，在放射性药物制备后的初始放射化学纯度不低于93％，优选不低于95％，更优选地不低于97％。在一个实施方案中，采用本申请方案的伊文思蓝衍生物药物水溶液在32℃，湿度60％RH的储存条件下，其API的放射化学纯度在48小时内保持不低于90％，优选地，在72小时内保持不低于90％。
采用本申请提供的放射性药物水溶液的制备方法具有如下有益效果：
本申请中，控制反应相溶液中龙胆酸的浓度为0.6-20.0mg/mL。低于0.6mg/mL时，龙胆酸的抗辐射分解作用不足，高于20.0mg/mL时，高浓度的龙胆酸将延缓反应动力学，不利地延长反应所需时间。该控制范围是在保证溶液稳定的前提下，尽量减少龙胆酸的浓度以避免对反应动力学的不利影响。
含有所述伊文思蓝衍生物分子的溶液与核素溶液混合前，先将核素溶液与含有第一稳定剂的溶液混合，经过给定时间后，再加入含有所述伊文思蓝衍生物分子的溶液，目的在于使第一稳定剂与核素溶液充分接触，淬灭核素溶液中存在的辐射分解带来的大量自由基，从而保护后加入反应体系的伊文思蓝衍生物分子不被活性自由基攻击，确保产物的初始放化纯。该工艺可使初始放化纯达到95.0-99.5％，而直接将含有所述伊文思蓝衍生物分子的溶液与核素溶液混合的合成工艺得到的产物的初始放化纯约为89％-93％。
采用本申请中的工艺方法至少能够保证标记率在90％以上，优选在95％以上，最优的在99％以上。在本申请提供的一些实施方案中，反应后不含有去除放射性杂质的纯化步骤，如制备液相分离、固相萃取分离等。若该放射性核素络合物溶液有无菌的要求，则在回收产物的步骤中还包含将所得溶液通过0.22μm除菌滤膜，还可进一步根据用量对该溶液进行进一步的稀释步骤。
具体实施方式
下面结合具体实施方式对本申请做进一步的详细描述，给出的实施例是为了能够更透彻地理解本发明，并且能够将本发明的范围完整的传达给本领域的技术人员。
需要说明的是，在说明书及权利要求当中使用了某些词汇来指称特定组件。本领域技术人员应可以理解，技术人员可能会用不同名词来称呼同一个组件。本说明书及权利要求并不以名词的差异作为区分组件的方式，而是以组件在功能上的差异作为区分的准则。如在通篇说明书及权利要求当中所提及的“包含”或“包括”为开放式用语，故应解释成“包含但不限定于”。说明书后续描述为实施本申请的较佳实施方式，然而所述描述乃以说明书的一般原则为目的，并非用以限定本申请的范围。本申请的保护范围当视所附权利要求所界定者为准。
本申请涉及一种放射性药物水溶液，其中包括经伊文思蓝(EB)片段修饰的靶向分子与放射性金属核素形成的络合物以及稳定剂。
在一个具体的实施方式中，所述经伊文思蓝(EB)片段修饰的靶向分子(或本申请说明书中所述“伊文思蓝衍生物分子”)为如式Ⅰ所示的化合物或其药学上可接受的酯、酰胺、溶剂化物、盐、或如式Ⅰ所示的化合物或其药学上可接受的酯的盐、或如式Ⅰ所示的化合物或其药学上可接受的酰胺的盐、或此如式Ⅰ所示的化合物或其药学上可接受的酯的溶剂化物、或如式Ⅰ所示的化合物或其药学上可接受的酰胺的溶剂化物、或如式Ⅰ所示的化合物或其药学上可接受的盐的溶剂化物。
其中，
L1是—(CH2)m—，其中m是0至12的整数，其中每个CH2可以单独地用—O—、—NH(CO)—、或—(CO)NH—替换，条件是没有两个相邻的CH2基团被替换；
L2是C1-C60连接基团，任选地包括—O—、—S—、—S(O)—、—S(O)2—、—N(R)—、—C(＝O)—、—C(＝O)O—、—OC(＝O)—、—N(R)C(＝O)—、—C(＝O)N(R)—、—OC(＝O)O—、—N(R)C(＝O)O—、—OC(＝O)N(R)—、 其中每个R为H或C1-C6烷基；
L3是—(CH2)n—，其中n是0至12的整数，其中每个CH2可以单独地用—O—、—NH(CO)—、或—(CO)NH—替换，条件是没有两个相邻的CH2基团被替换；
Ch是螯合基团；
Tg是靶向基团。
伊文思蓝(Evans Blue,EB)是一种非膜渗透性的偶氮染料制剂，因其在血液中与血清白蛋白有很高的亲和力,因此利用其与白蛋白可逆结合的性质，以截短的EB片段(truncated Evans Blue,tEB)修饰靶向分子，使靶向分子能够通过tEB片段与内源性血清白蛋白进行可逆的结合，将血清白蛋白作为药物分子的可逆载体，以延长药物分子在血液中的半衰期，增加药物分子可用性，进一步增加药物分子在肿瘤中的积累与保留时间。伊文思蓝(Evans Blue,EB)染料的结构式如式Ⅵ所示：
在本申请一些具体的实施方式中，伊文思蓝衍生物分子为式Ⅰ化合物。在另一些具体的实施方式中，伊文思蓝衍生物分子为式Ⅰ化合药学上可接受的酯、酰胺、溶剂化物或盐。在另一些具体的实施方式中，伊文思蓝衍生物分子为式Ⅰ化合药学上可接受的酯的盐。在另一些具体的实施方式中，伊文思蓝衍生物分子为式Ⅰ化合药学上可接受的酰胺的盐。在另一些具体的实施方式中，伊文思蓝衍生物分子为式Ⅰ化合药学上可接受的酯的溶剂化物。在另一些具体的实施方式中，伊文思蓝衍生物分子为式Ⅰ化合药学上可接受的酰胺的溶剂化物。在另一些具体的实施方式中，伊文思蓝衍生物分子为式Ⅰ化合药学上可接受盐的溶剂化物。所述伊文思蓝衍生物分子可以通过常规化学方法从含有碱性或酸性部分的母体化合物合成。
在一个具体的实施方式中，式Ⅰ中的L1为—NH(CO)—，L3为—NH(CO)CH2—，即式Ⅰ的化合物为式Ⅶ的化合物：
在一个具体的实施方式中，式Ⅰ中的螯合基团Ch选自
优选的，式Ⅰ中的螯合基团Ch为
螯合基团具有两个或两个以上的配位原子并能与同一中心原子结合形成环状结构的基团，其可以与通常为金属离子的单个中心原子形成两个或更多个单独的配位键。本申请中的螯合基团是具有多个N，O或S杂原子的有机基团，并且具有允许两个或更多个杂原子与相同金属离子形成键的结构。在本申请的具体实施方式中，所述螯合基团用于与放射性金属核素形成键的结构。
在本申请一个具体的实施方式中，式Ⅰ中的靶向基团Tg为能够特异性靶向某一生物靶点的化合基团。在一些实施方案中，Tg选自能够靶向生长抑素受体(SSTR)、前列腺特异性膜抗原(PSMA)、成纤维细胞活化蛋白 (FAP)、叶酸受体(FR)、表皮生长因子受体或整合素的化合基团。在一些具体的实施方式中，靶向基团Tg选自
在本申请一个具体的实施方式中，式Ⅰ的化合物为EB-PSMA，结构式如式Ⅱ所示：
在本申请一个具体的实施方式中，式Ⅰ的化合物为DOTA-EB-TATE，结构式如式Ⅲ所示：
在本申请一个具体的实施方式中，式Ⅰ的化合物为EB-FAPI，结构式如式Ⅳ所示：
在本申请一个具体的实施方式中，式Ⅰ的化合物为NMEB-RGD，结构式如式Ⅴ所示：
在本申请的具体实施方式中，与伊文思蓝衍生物分子形成络合物的放射性金属核素选自177Lu、99mTc、68Ga、64Cu、67Cu、111In、86Y、90Y、89Zr、186Re、188Re、153Sm、82Rb、166Ho、225Ac、212Pb、213Bi、212Bi、227Th。所述放射性金属核素可以通过螯合，或通过其它方式，如化学领域中已知的常 规共价键或离子键与螯合基团Ch结合。放射性核素可以是合适的目的，例如放射治疗和/或诊断。
在本申请一个具体的实施方式中，所述放射性金属核素在药物水溶液制剂中以0.037-1850MBq/mL的体积放射性浓度存在。
在本申请的一个具体的实施方式中，所述放射性药物水溶液中的稳定剂为抗放射性分解降解稳定剂，具体的，所述稳定剂选自龙胆酸及其盐、抗坏血酸及其盐、组氨酸、半胱氨酸及其盐、甲硫氨酸、硒甲硫氨酸、硫代硫酸盐、麦芽糖、肌醇、苯甲醇、海藻糖、聚维酮、烟酰胺、乙醇、姜黄素、褪黑素中的一种或两种以上，优选为龙胆酸、乙醇、甲硫氨酸中的一种或两种以上。
在一个具体的实施方式中，所述稳定剂在药物水溶液中的总浓度为0.5-400mg/mL，例如可以为0.5、1、2、5、10、25、50、100、150、200、250、300、350、400mg/mL。优选的，为1-80mg/mL，例如可以为1、2、5、10、15、20、25、30、35、40、45、50、55、60、65、70、75、80mg/mL。
在一个具体的实施方式中，所述稳定剂在形成核素络合物的络合反应期间以及反应结束后分别加入。其中，所述在络合反应期间加入是指，所述稳定剂与形成络合物的放射性核素溶液和伊文思蓝衍生物分子溶液在达成足以发生络合反应的条件时共同组成反应相溶液；所述在反应结束后加入是指，发生所述络合反应一定时间、络合物已形成完成后所加入稳定剂。进一步的，在所述络合反应期间加入的稳定剂为第一稳定剂，在反应结束后加入的稳定剂为第二稳定剂。第一稳定剂一般为具有抗氧化性质的小分子化合物，用以在高辐射下降低辐射分解。第二稳定剂的主要作用是维持制剂在储存期间的稳定性。第一稳定剂和第二稳定剂可以选自相同的稳定剂，也可以选自不同的稳定剂。
在一个具体的实施方式中，第一稳定剂选自龙胆酸及其盐、抗坏血酸及其盐、组氨酸、半胱氨酸及其盐、甲硫氨酸、硒甲硫氨酸、硫代硫酸盐、麦芽糖、肌醇、苯甲醇、海藻糖、聚维酮、烟酰胺、乙醇、姜黄素、褪黑素中的一种或两种以上，优选为龙胆酸。
在一个具体的实施方式中，第二稳定剂选自龙胆酸及其盐、抗坏血酸及其盐、组氨酸、半胱氨酸及其盐、甲硫氨酸、硒甲硫氨酸、硫代硫酸盐、麦芽糖、肌醇、苯甲醇、海藻糖、聚维酮、烟酰胺、乙醇、姜黄素、褪黑 素中的一种或两种以上，优选为龙胆酸、乙醇或甲硫氨酸。
在一个优选的实施方式中，所述第一稳定剂和第二稳定剂相同，均选自龙胆酸或其盐。其中，在络合发应过程中加入龙胆酸(即第一稳定剂)，其在反应体系中的浓度范围为0.6-20mg/mL，优选为2-10mg/mL，例如可以为2、2.5、3、3.5、4、4.5、5、5.5、6、6.5、7、7.5、8、8.5、9、9.5、10mg/mL。在反应结束后的制剂中继续加入龙胆酸(即第二稳定剂)，使龙胆酸在整个药物水溶液中以0.1-10mg/mL的总浓度存在，优选为0.5-5mg/mL，，例如可以为0.5、0.8、1.0、1.2、1.5、2.0、2.3、2.4、2.5、2.6、2.7、2.8、3.0、3.2、3.5、3.8、4.0、4.5、5.0mg/mL。
在另一些优选的实施方式中，所述稳定剂为两种不同的稳定剂。
在一个具体的实施方式中，在所述络合反应期间的反应体系中加入的第一稳定剂为龙胆酸或其盐。其在药物水溶液中以0.5-5mg/mL的浓度存在，优选为0.5-2mg/mL，例如可以为0.5、0.8、1.0、1.2、1.5、1.8、2.0mg/mL。在反应结束后加入的第二稳定剂为乙醇，其在药物水溶液中以0-400mg/mL的浓度存在，优选为10-120mg/mL，例如可以为10、30、50、60、70、80、100、120mg/mL。其体积分数为0％-50％，优选为1％-15％，例如可以为1％、3％、5％、6％、7％、8％、9％、10％、12％、15％。
在一个具体的实施方式中，在所述络合反应期间的反应体系中加入的第一稳定剂为龙胆酸或其盐，其在药物水溶液中以0.5-5mg/mL的浓度存在，优选为0.5-2mg/mL，例如可以为0.5、0.8、1.0、1.2、1.3、1.4、1.5、1.6、1.8、2.0mg/mL。在反应结束后加入的第二稳定剂为L-甲硫氨酸，其在药物水溶液中以0-50mg/mL的浓度存在，优选为1-10mg/mL，例如可以为1、2、3、4、5、6、7、8、9、10mg/mL。
在另一些具体的实施方式中，两种稳定剂优选不含有抗坏血酸及其盐。
在一个具体的实施方式中，所述药物水溶液中还包括缓冲液。所述缓冲液可以在所述络合反应期间加入，用以调节反应相溶液的pH，也可以在反应结束后再次加入用以调节制剂溶液的pH。两次所加入的缓冲液可以一样也可以不一样。缓冲液可以选用醋酸盐体系(如醋酸-醋酸钠体系、醋酸钠体系)、柠檬酸盐体系(如柠檬酸-柠檬酸钠体系)、磷酸盐体系(如磷酸二氢钠-磷酸氢二钠体系)、甲酸盐体系(如甲酸-甲酸钠体系)的溶液。在一个优选的实施方式中，在反应相溶液中缓冲盐的浓度为0.01-2.0M。 在一个优选的实施方式中，在最终药物水溶液中总的缓冲盐浓度为0.005-0.5M。
在一个具体的实施方式中，所述药物水溶液中还包括助溶剂，其作用在于降低API在各接触材料表面(尤其是玻璃和塑料表面)的吸附。所述助溶剂选自聚山梨酯20、聚山梨酯40、聚山梨酯60、聚山梨酯80、泊洛沙姆188、聚氧乙烯蓖麻油、司盘、乙醇、丙二醇、丙三醇、聚乙二醇(平均分子量为200～8000)、山梨醇、二甲基亚砜、十二烷基硫酸钠中的一种或两种以上，优选为聚山梨酯80。在一个具体的实施方式中，所述助溶剂在药物水溶液中的浓度为0.01-10mg/mL，优选0.05-1.0mg/mL，例如可以为0.05、0.08、0.1、0.2、0.3、0.4、0.5、0.6、0.7、0.8、1.0mg/mL。
在一个具体的实施方式中，所述的药物水溶液中还包括游离金属核素的螯合剂。所述螯合剂的作用为与药物水溶液中未反应的游离核素离子发生络合，以减少游离的放射性核素离子在体内对健康组织产生不必要的照射。因而要求该螯合剂须具备较强的与核素离子络合的能力，即便在注射液进入生物体内被血浆稀释后，仍能在较低的浓度条件下与游离的核素离子迅速反应，该络合反应需要迅速，条件温和，在室温条件下就可完全进行。在一个具体的实施方式中，所述螯合剂为喷替酸或其盐，优选为喷替酸。所述螯合剂在药物水溶液中的浓度为0.005-0.1mg/mL，例如可以为0.005、0.01、0.02、0.03、0.04、0.05、0.06、0.07、0.08、0.09、0.1mg/mL。在该范围内，喷替酸对游离核素离子有充分的络合能力，并且该络合物在辐射分解作用下也能至少能保持至少48h的稳定性，更优的能够保持72h稳定，即不会由于该螯合剂的辐射分解而造成游离核素离子的释放。
本申请还提供了将所述药物水溶液应用于肿瘤的放射治疗和/或诊断中的方案，包括通过向患者施用有效量的所述药物水溶液，或与其他一种或多种肿瘤免疫学治疗剂共同组成的组合物。在一些具体的实施方式中，所述药物水溶液或包含其的组合物可用于治疗神经内分泌瘤、前列腺癌、乳腺癌、卵巢癌、胰腺癌、肝癌、肺癌、结直肠癌、黑色素瘤等。在另一些具体的实施方式中，本申请提供的药物水溶液或包含其的组合物还可用于制备预防或治疗糖尿病、阿尔茨海默病的药物中。
在一个具体的实施方式中，采用本申请所提供的药物水溶液，至少能够使在32℃，湿度60％RH储存条件下，API的放射化学纯度在48h内不 低于90％，更优地在72h内放射化学纯度不低于90％，所述放射化学纯度为通过HPLC测定的数值。
本申请还涉及一种放射性药物水溶液的制备方法，在一个具体的实施方式中，包括如下步骤：
将含有第一稳定剂的溶液与含有放射性核素的溶液在反应容器中混合；
给定时间后，将含有所述伊文思蓝衍生物分子的溶液加入至所述反应容器，优选所述给定时间为0.1分钟～20分钟，进一步优选3分钟～10分钟；
所述伊文思蓝衍生物分子与放射性核素发生反应得到所述放射性核素络合物；
反应给定时间后向所述反应容器中加入含有第二稳定剂的溶液；
回收得到的所述放射性药物水溶液；
其中，所述伊文思蓝衍生物分子为如式Ⅰ所示的化合物或其药学上可接受的酯、酰胺、溶剂化物、盐、或如式Ⅰ所示的化合物或其药学上可接受的酯的盐、或如式Ⅰ所示的化合物或其药学上可接受的酰胺的盐、或此如式Ⅰ所示的化合物或其药学上可接受的酯的溶剂化物、或如式Ⅰ所示的化合物或其药学上可接受的酰胺的溶剂化物、或如式Ⅰ所示的化合物或其药学上可接受的盐的溶剂化物，
其中，
L1是—(CH2)m—，其中m是0至12的整数，其中每个CH2可以单独地用—O—、—NH(CO)—、或—(CO)NH—替换，条件是没有两个相邻的CH2基团被替换；
L2是C1-C60连接基团，任选地包括—O—、—S—、—S(O)—、—S(O)2—、—N(R)—、—C(＝O)—、—C(＝O)O—、—OC(＝O)—、—N(R)C(＝O)—、—C(＝O)N(R)—、—OC(＝O)O—、—N(R)C(＝O)O—、—OC(＝O)N(R)—、 其中每个R为H或C1-C6烷基；
L3是—(CH2)n—，其中n是0至12的整数，其中每个CH2可以单独地用—O—、—NH(CO)—、或—(CO)NH—替换，条件是没有两个相邻的CH2基团被替换；
Ch是螯合基团；
Tg是靶向基团。
在本申请的一个实施方式中，所述放射性药物水溶液包含放射性核素络合物，所述放射性核素络合物是由放射性核素与所述伊文思蓝衍生物分子形成的。
在本申请具体的实施方式中，所述含有放射性核素的溶液为含有放射性金属元素的溶液，在一些具体的实施方式中，所述放射性核素选自177Lu、99mTc、68Ga、64Cu、67Cu、111In、86Y、90Y、89Zr、186Re、188Re、153Sm、82Rb、166Ho、225Ac、212Pb、213Bi、212Bi或227Th。在一个具体的实施方式中，所述放射性核素为177Lu，且在与伊文思蓝衍生物分子发生络合反应的步骤中，所述放射性核素的比活度不低于20Ci/mg，优选为不低于60Ci/mg，最优选为不低于80Ci/mg。过低比活度的放射性核素会影响放射性标记效率。
在本申请中，所述伊文思蓝衍生物分子为以截短的伊文思蓝片段(truncated Evans Blue,tEB)修饰的靶向分子。在一些具体的实施方式中，伊文思蓝衍生物分子为式Ⅰ化合物。在另一些具体的实施方式中，伊文思蓝衍生物分子为式Ⅰ化合药学上可接受的酯、酰胺、溶剂化物或盐。在另一些具体的实施方式中，伊文思蓝衍生物分子为式Ⅰ化合药学上可接受的酯的盐。在另一些具体的实施方式中，伊文思蓝衍生物分子为式Ⅰ化合药学上可接受的酰胺的盐。在另一些具体的实施方式中，伊文思蓝衍生物分子为式Ⅰ化合药学上可接受的酯的溶剂化物。在另一些具体的实施方式中，伊文思蓝衍生物分子为式Ⅰ化合药学上可接受的酰胺的溶剂化物。在另一些具体的实施方式中，伊文思蓝衍生物分子为式Ⅰ化合药学上可接受盐的溶剂化物。
在一个具体的实施方式中，式Ⅰ中的L1为—NH(CO)—，L3为—NH(CO)CH2—，即式Ⅰ的化合物为式Ⅶ的化合物：
在一个具体的实施方式中，式Ⅰ中的螯合基团Ch选自
优选的，式Ⅰ中的螯合基团Ch为
在本申请一个具体的实施方式中，式Ⅰ中的靶向基团Tg为能够特异性靶向某一生物靶点的化合基团。在一些实施方案中，Tg选自能够靶向生长抑素受体(SSTR)、前列腺特异性膜抗原(PSMA)、成纤维细胞活化蛋白(FAP)、叶酸受体(FR)、表皮生长因子受体或整合素的化合基团。在 一些具体的实施方式中，靶向基团Tg选自
在本申请一个具体的实施方式中，式Ⅰ的化合物为EB-PSMA，结构式如式Ⅱ所示：
在本申请一个具体的实施方式中，式Ⅰ的化合物为DOTA-EB-TATE，结构式如式Ⅲ所示：

在本申请一个具体的实施方式中，式Ⅰ的化合物为EB-FAPI，结构式如式Ⅳ所示：
在本申请一个具体的实施方式中，式Ⅰ的化合物为NMEB-RGD，结构式如式Ⅴ所示：
在本申请一个具体的实施方式中，所述含有放射性核素的溶液是从原料瓶中取出后加入至所述反应容器中，并且在所述含有放射性核素的溶液被取出后，以冲洗液对原料瓶进行冲洗，以提取原料瓶中残留的核素溶液，并将冲洗后的溶液转入所述反应容器中与所述含有放射性核素的溶液混合。
在一个具体的实施方式中，所述冲洗液为水溶液，优选地选自含有第一稳定剂的溶液、含有缓冲盐的溶液、水或氯化钠注射液。
在一个优选的实施方式中，所述冲洗液选自注射用水或氯化钠注射液。
在一个优选的实施方式中，使用所述冲洗液重复所述冲洗一次或更多次。
在本申请一个具体的实施方式中，所述第一稳定剂选自龙胆酸及其盐、抗坏血酸及其盐、组氨酸、半胱氨酸及其盐、甲硫氨酸、硒甲硫氨酸、硫代硫酸盐、麦芽糖、肌醇、苯甲醇、海藻糖、聚维酮、烟酰胺、乙醇、姜黄素、褪黑素中的一种或两种以上，优选为龙胆酸。
在本申请一个具体的实施方式中，将含有第一稳定剂的溶液与含有放射性核素的溶液在反应容器中混合，经过给定时间后，再将含有所述伊文思蓝衍生物分子的溶液加入至所述反应容器。所述给定时间可使第一稳定剂与核素溶液充分接触，淬灭其中存在的辐射分解带来的大量自由基，从而保护随后加入反应体系的伊文思蓝衍生物分子不被活性自由基攻击，利于提高最终产物的初始放化纯。
在一个优选的实施方式中，所述给定时间为0.1分钟～20分钟，例如可以为0.1、0.2、0.3、0.5、0.8、1、1.5、2、2.5、3、3.5、4、4.5、5、5.5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20分钟；进一步优选为3分钟～10分钟，例如可以为3、3.5、4、4.5、5、5.5、6、6.5、7、7.5、8、8.5、9、9.5、10分钟。
在一个具体的实施方式中，将所述含有伊文思蓝衍生物分子的溶液加入至反应相溶液中与放射性核素发生反应得到所述放射性核素络合物。
在一个具体的实施方式中，所述伊文思蓝衍生物分子(标记前体)溶液选自浓度为0.05-10.0mg/mL的化合物溶液，其制备方法为将标记前体的冻干粉溶解于灭菌注射用水或乙醇中。
在一个优选的实施方式中，所述放射性核素络合物为177Lu-DOTA-EB-TATE。
在一个具体的实施方式中，所述伊文思蓝衍生物分子与放射性核素发生反应得到所述放射性核素络合物的反应过程中，存在的第一稳定剂为龙胆酸，其在上述反应相中的浓度为0.6-20.0mg/mL，优选为2-10mg/mL，例如可以为2、2.5、3、3.5、4、4.5、5、5.5、6、6.5、7、7.5、8、8.5、9、9.5、10mg/mL。最优选为3.0-5.0mg/mL，例如可以为3.0、3.2、3.4、3.5、3.6、3.8、4.0、4.2、4.5、4.8、5.0mg/mL。当龙胆酸在反应相体系中的浓度超过了控制范围，会极大地减慢反应速率，不利于整个合成过程；而龙胆酸浓度低于控制浓度时，则会由于稳定剂浓度的不足而使辐射降解杂质增加。
在一个具体的实施方式中，所述伊文思蓝衍生物分子与放射性核素发生反应所形成的反应相溶液中，所述伊文思蓝衍生物分子与放射性核素之间的摩尔比为1.5–50，优选为5–20，例如可以为5、8、10、12、15、18、20。所述摩尔比指反应体系中伊文思蓝衍生物分子(标记前体)与放射性核素的摩尔量之比。在反应相溶液中，摩尔比的提高有利于放射性核素的完全反应，使标记率提高，但未被标记的标记前体会在生物体内与API产生竞争。而过低的摩尔比使API缺乏载体，在生物体内容易被其他非特异性靶点结合而损失，达不到预想的治疗或诊断效果。
在本申请的实施方式中，还可以对所述反应相溶液中反应相的浓度进行控制。理论上，反应相浓度越高，标记反应速率越快，但同时由放射性核素引起的辐射分解效应也越强，因此反应相浓度不能过高，而过低的反应相浓度则使反应体积变大，限制大批量的核素络合物生产。对本申请的制备方法而言，在反应相溶液中，所述伊文思蓝衍生物分子浓度在0.01-1.0mg/mL范围内，优选0.05-0.5mg/mL，例如可以为0.05、0.1、0.15、0.2、0.25、0.3、0.35、0.4、0.45、0.5mg/mL。
在本申请的实施方式中，所述伊文思蓝衍生物分子与放射性核素发生络合反应的步骤中，对反应温度与时间进行控制，以此来控制达到反应标记率>90％，化学纯>90％，放化纯>90％。在一个具体的实施方式中，所述反应温度为50-100℃，优选为60-80℃，例如可以为60、62、65、68、70、72、75、78、80℃；反应时间为5-60分钟，例如可以为5、10、12、15、18、20、25、30、40、50、60分钟，优选为10-30分钟，最优选10-20分钟，例如可以为10、11、12、13、14、15、16、17、18、19、20分钟。
在一个具体的实施方式中，所述伊文思蓝衍生物分子与放射性核素经过上述反应时间以形成络合物反应结束后，加入所述第二稳定剂。具体的，所述第二稳定剂选自龙胆酸及其盐、抗坏血酸及其盐、组氨酸、半胱氨酸及其盐、甲硫氨酸、硒甲硫氨酸、硫代硫酸盐、麦芽糖、肌醇、苯甲醇、海藻糖、聚维酮、烟酰胺、乙醇、姜黄素、褪黑素中的一种或两种以上，优选为龙胆酸、乙醇或甲硫氨酸。
在一个具体的实施方式中，在所述放射性药物水溶液中，第二稳定剂的浓度为0-400mg/mL，例如可以为0、0.5、1、2、5、10、25、50、100、150、200、250、300、350、400mg/mL。
在本申请的实施方式中，所述制备方法还包括在所述伊文思蓝衍生物分子与放射性核素发生反应之前加入缓冲盐溶液，优选地，所述缓冲盐溶液存在于所述含有第一稳定剂的溶液中。
在一个具体的实施方式中，所述缓冲盐溶液选自醋酸盐、柠檬酸盐、磷酸盐或甲酸盐溶液，优选为醋酸-醋酸钠缓冲盐溶液。
缓冲盐溶液的加入可以调节反应体系的pH值，控制反应相体系的pH值在3.5-6.0范围内，例如可以为，3.5、3.8、4、4.2、4.4、4.6、4.8、5、5.5、6，优选pH值为3.5-5。在一个具体的实施方式中，控制最终制剂溶液的pH值为4–6，例如可以为4、4.2、4.5、4.8、5、5.2、5.5、5.8、6。
在一个具体的实施方式中，在反应给定时间后向所述反应容器中加入含有第二稳定剂的溶液的步骤还包括向所述反应容器中加入助溶剂。
在一个具体的实施方式中，所述助溶剂选自聚山梨酯20、聚山梨酯40、聚山梨酯60、聚山梨酯80、泊洛沙姆188、聚氧乙烯蓖麻油、司盘、乙醇、丙二醇、丙三醇、聚乙二醇(平均分子量为200～8000)、山梨醇、二甲基亚砜、十二烷基硫酸钠中的一种或两种以上，优选为聚山梨酯80。在一个具体的实施方式中，加入所述助溶剂使其在药物水溶液中的浓度为0.01-10mg/mL，优选为0.05-1.0mg/mL、例如可以为0.05、0.08、0.1、0.2、0.3、0.4、0.5、0.6、0.7、0.8、1.0mg/mL。
在一个具体的实施方式中，在反应给定时间后向所述反应容器中加入含有第二稳定剂的溶液的步骤还包括向所述反应容器中加入游离核素螯合剂，所述螯合剂可选自喷替酸及其盐，优选为喷替酸。在一个优选的实施方式中，加入所述螯合剂使其在药物水溶液中的浓度为0.005-0.1mg/mL，例如可以为0.005、0.01、0.02、0.03、0.04、0.05、0.06、0.07、0.08、0.09、0.1mg/mL。
在一个具体的实施方式中，本申请的制备方法还包括对放射性药物水溶液进行过滤除菌，在一个具体的实施方式中，使放射性药物水溶液通过0.22μm滤膜过滤除菌。
在一个具体的实施方式中，本申请的制备方法还包括对所述放射性水溶液进行稀释，优选地，加入氯化钠注射液进行稀释以进行回收。
在一个优选的实施方式中，所述过滤除菌和稀释在加入含有第二稳定剂的溶液后进行。本申请中并不限定所述过滤除菌和稀释步骤的先后顺序， 即可以先进行过滤除菌再进行稀释，或者先进行稀释，再通过滤膜过滤除菌，随后进行回收。
在一个具体的实施方式中，本申请提供了一种以如下顺序制备177Lu-DOTA-EB-TATE放射性药物水溶液的方法：
a.将含有500mCi 177Lu和盐酸的核素溶液从原料瓶转移至反应瓶中；
b.将含有2.0M甲酸-甲酸钠缓冲盐和50mg/mL龙胆酸的冲洗液1mL加入上述原料瓶中，用以冲洗原料瓶中残留的177Lu溶液；
c.将冲洗后原料瓶中的混合溶液一并转入反应瓶中；
d.用3mL注射用水加入上述原料瓶中，用以冲洗原料瓶；
e.将冲洗后原料瓶中的混合溶液一并转入反应瓶中；
f.将含有上述溶液的反应瓶于室温下静置10分钟；
g.将0.5mL DOTA-EB-TATE溶液加入反应瓶中；
h.将该反应瓶升温至90℃，反应15分钟；
i.反应结束后冷却反应瓶，向反应瓶中加入含有0.5mg/mL喷替酸、45mg/mL龙胆酸、2.0mg/mL聚山梨酯80的混合溶液10mL；
j.将所得溶液通过0.22μm滤膜过滤除菌；
k.用35mL氯化钠注射液稀释所得溶液；
l.回收所得产物。
实施例
下述实施例中所使用的实验方法如无特殊要求，均为常规方法。
下述实施例中使用的前体EB-PSMA按照文献方法(Bioconjugate Chem.2018,29,3213-3221)合成得到。
下述实施例中使用的前体DOTA-EB-TATE按照文献方法(Theranostics.2018；8:735-745)合成得到。
下述实施例中使用的前体EB-FAPI按照文献方法(Theranostics.2022；12(1):422-433)合成得到。
下述实施例中所使用的龙胆酸购自成都普瑞法科技开发有限公司，喷替酸购自江西阿尔法高科药业有限公司。
其他材料、试剂等，如无特殊说明，均可从商业途径得到。
实施例1：药物水溶液中稳定剂的选择
处方(1)：[177Lu]Lu-DOTA-EB-TATE药物水溶液的制备
反应相溶液的配制：在反应容器中加入10mCi无载体氯化镥[177Lu]溶液(约10μL)、20μL甲酸-甲酸钠缓冲盐溶液(其中含有50mg/mL龙胆酸)、60μL注射用水，混合均匀后，将此混合溶液在室温下静置3分钟。然后继续向该反应容器中加入10μL DOTA-EB-TATE前体溶液，并混合均匀，该混合溶液为反应相溶液。
加热反应与冷却：将上述反应相溶液置于预热至90℃的加热器中反应15分钟，反应结束后取出反应容器冷却15分钟。
配辅与稀释：在反应相溶液冷却至室温后，向反应容器中加入200μL配辅溶液，配辅溶液中含有45mg/mL龙胆酸、0.5mg/mL喷替酸和2.0mg/mL聚山梨酯80。最后向反应容器中加入氯化钠注射液将总体积稀释至1.0mL，得到最终制剂溶液。
最终制剂溶液中含有10mg/mL龙胆酸、0.1mg/mL喷替酸、0.4mg/mL聚山梨酯80，其中API分子[177Lu]Lu-DOTA-EB-TATE在校准时刻的活度浓度为10mCi/mL，校准时刻是指生产结束时间(T0)。将制剂溶液储存于稳定性箱中，储存温度设置为32℃，储存湿度设置为60％RH。
在T0时刻使用Radio-HPLC检测制剂溶液的放射化学纯度为100％，在T0时刻使用ITLC检测制剂溶液的放射化学纯度为100％。
在T0+48h时刻取出稳定箱中的制剂溶液150μL用于稳定性检验。使用Radio-HPLC检测制剂溶液的放射化学纯度为94％，使用ITLC检测制剂溶液的放射化学纯度为100％。
在T0+72h时刻取出稳定箱中的制剂溶液150μL用于稳定性检验。使用Radio-HPLC检测制剂溶液的放射化学纯度为92％，使用ITLC检测制剂溶液的放射化学纯度为100％。
处方(2)：[177Lu]Lu-DOTA-EB-TATE药物水溶液的制备
反应相溶液的配制：在反应容器中加入10mCi无载体氯化镥[177Lu]溶液(约10μL)、20μL甲酸-甲酸钠缓冲盐溶液(其中含有50mg/mL龙胆酸)、160μL注射用水，混合均匀后，将此混合溶液在室温下静置3分钟。然后继续向该反应容器中加入10μL DOTA-EB-TATE前体溶液，并混合均匀，该混合溶液为反应相溶液。
加热反应与冷却：将上述反应相溶液置于预热至65℃的加热器中反应40分钟，反应结束后取出反应容器冷却15分钟。
配辅与稀释：在反应相溶液冷却至室温后，向反应容器中加入100μL 配辅溶液，配辅溶液中含有0.3mg/mL喷替酸和1.0mg/mL聚山梨酯80。再加入无水乙醇50mg，最后向反应容器中加入氯化钠注射液将总体积稀释至1.0mL，得到最终制剂溶液。
最终制剂溶液中含有1mg/mL龙胆酸、50mg/mL乙醇、0.03mg/mL喷替酸、0.1mg/mL聚山梨酯80，其中API分子[177Lu]Lu-DOTA-EB-TATE在校准时刻的活度浓度为10mCi/mL，校准时刻是指生产结束时间(T0)。将制剂溶液储存于稳定性箱中，储存温度设置为32℃，储存湿度设置为60％RH。
在T0时刻使用Radio-HPLC检测制剂溶液的放射化学纯度为100％，在T0时刻使用ITLC检测制剂溶液的放射化学纯度为100％。
在T0+48h时刻取出稳定箱中的制剂溶液150μL用于稳定性检验。使用Radio-HPLC检测制剂溶液的放射化学纯度为93％，使用ITLC检测制剂溶液的放射化学纯度为100％。
在T0+72h时刻取出稳定箱中的制剂溶液150μL用于稳定性检验。使用Radio-HPLC检测制剂溶液的放射化学纯度为92％，使用ITLC检测制剂溶液的放射化学纯度为99％。
处方(3)：[177Lu]Lu-EB-FAPI药物水溶液的制备
反应相溶液的配制：在反应容器中加入20mCi无载体氯化镥[177Lu]溶液(约20μL)、20μL醋酸铵缓冲盐溶液(其中含有50mg/mL龙胆酸)、40μL注射用水，混合均匀后，将此混合溶液在室温下静置10分钟。然后继续向该反应容器中加入20μL EB-FAPI前体溶液，并混合均匀，该混合溶液为反应相溶液。其中前体EB-FAPI为式Ⅳ的化合物，R＝H。
加热反应与冷却：将上述反应相溶液置于预热至95℃的加热器中反应30分钟，反应结束后取出反应容器冷却15分钟。
配辅与稀释：在反应相溶液冷却至室温后，向反应容器中加入500μL配辅溶液，配辅溶液中含有20mg/mL甲硫氨酸、4.0mg/mL龙胆酸、0.2mg/mL喷替酸和0.8mg/mL聚山梨酯80。最后向反应容器中加入氯化钠注射液将总体积稀释至1.0mL，得到最终制剂溶液。
最终制剂溶液中含有3.0mg/mL龙胆酸、10mg/mL甲硫氨酸、0.1mg/mL喷替酸、0.4mg/mL聚山梨酯80，其中API分子[177Lu]Lu-EB-FAPI在校准时刻的活度浓度为20mCi/mL，校准时刻是指生产结束时间(T0)。将制剂溶液储存于稳定性箱中，储存温度设置为32℃，储存湿度设置为60％RH。
在T0时刻使用Radio-HPLC检测制剂溶液的放射化学纯度为98％，在T0时刻使用ITLC检测制剂溶液的放射化学纯度为100％。
在T0+48h时刻取出稳定箱中的制剂溶液150μL用于稳定性检验。使用Radio-HPLC检测制剂溶液的放射化学纯度为96％，使用ITLC检测制剂溶液的放射化学纯度为100％。
在T0+72h时刻取出稳定箱中的制剂溶液150μL用于稳定性检验。使用Radio-HPLC检测制剂溶液的放射化学纯度为93％，使用ITLC检测制剂溶液的放射化学纯度为100％。
处方(4)：[177Lu]Lu-EB-PSMA药物水溶液的制备
反应相溶液的配制：在反应容器中加入10mCi无载体氯化镥[177Lu]溶液(约10μL)、20μL醋酸-醋酸钠缓冲盐溶液(其中含有50mg/mL龙胆酸)、60μL注射用水，混合均匀后，将此混合溶液在室温下静置3分钟。然后继续向该反应容器中加入10μL EB-PSMA前体溶液，并混合均匀，该混合溶液为反应相溶液。
加热反应与冷却：将上述反应相溶液置于预热至80℃的加热器中反应15分钟，反应结束后取出反应容器冷却15分钟。
配辅与稀释：在反应相溶液冷却至室温后，向反应容器中加入100μL配辅溶液，配辅溶液中含有30mg/mL抗坏血酸、1.0mg/mL喷替酸和4.0mg/mL聚山梨酯80。最后向反应容器中加入氯化钠注射液将总体积稀释至1.0mL，得到最终制剂溶液。
最终制剂溶液中含有1.0mg/mL龙胆酸、3.0mg/mL抗坏血酸、0.1mg/mL喷替酸、0.4mg/mL聚山梨酯80，其中API分子[177Lu]Lu-EB-PSMA在校准时刻的活度浓度为10mCi/mL，校准时刻是指生产结束时间(T0)。将制剂溶液储存于稳定性箱中，储存温度设置为32℃，储存湿度设置为60％RH。
在T0时刻使用Radio-HPLC检测制剂溶液的放射化学纯度为93％。
在T0+48h时刻取出稳定箱中的制剂溶液150μL，用Radio-HPLC检测制剂溶液的放射化学纯度为81％。
在T0+72h时刻取出稳定箱中的制剂溶液150μL，用Radio-HPLC检测制剂溶液的放射化学纯度为66％。
实验结果分析：采用如上述处方(1)～(3)中以龙胆酸、龙胆酸和乙醇、龙胆酸和甲硫氨酸作为稳定剂制备的药物水溶液，在48h和72h均能得到90％以上的API放化纯，明显优于处方(4)中采用的抗坏血酸的稳定剂选择。这表明本申请发明内容所提供的药物水溶液能保持更佳的稳定性。 实施例2反应温度与时间的控制
反应相溶液的配制：与处方(3)相同。
加热反应与冷却：将上述反应相溶液置于室温或已预热至不同温度的加热器中反应120分钟。考察的温度包含室温、50℃、60℃、70℃、80℃、90℃、95℃、100℃。
标记率检测：在反应进行至不同时间点时，取出5μCi反应相溶液进行ITLC检测，取样前后反应相溶液均置于加热器中反应，即取样过程不影响反应的持续进行。计算标记率(ITLC)＝与前体分子螯合的放射性活度÷总放射性活度，考察的时间包括1、5、10、30、45、60、90、120分钟。
对标记率而言，反应温度与反应时间是两个互补的工艺条件，“在较低温度下反应较长时间”或“在较高温度下反应较短时间”均能使反应标记率(ITLC)≥99％，此时我们认为标记反应已完全进行。但考虑到高温会促使化学杂质和放射化学杂质生成，而大于60分钟的反应时间则不利于工艺过程的控制，因此选择将反应温度控制在50-100℃，将反应时间控制在5-60分钟，优选的，将反应温度控制在60-80℃，将反应时间控制在10-30分钟。实施例3投料比例的控制
反应相溶液的配制：在反应容器中加入10mCi无载体氯化镥[177Lu]溶液(约10μL)和20μL甲酸-甲酸钠缓冲盐溶液(其中含有50mg/mL龙胆酸)，将此混合溶液在室温下静置3分钟。然后继续向该反应容器中加入不同体积的注射用水与DOTA-EB-TATE前体溶液，使反应相中DOTA-EB-TATE与氯化镥[177Lu]的投料摩尔比分别为1、1.5、2、3、5、10、15、30、50，并使溶液总体积为0.1mL，将溶液混合均匀，该混合溶液为反应相溶液。
加热反应与冷却：与处方(1)相同。
配辅与稀释：与处方(1)相同。
标记率检测：取5μCi反应相溶液进行ITLC检测，计算标记率(ITLC)＝与前体分子螯合的放射性活度÷总放射性活度。
当前体分子与核素的投料摩尔比为1.5-50时，其标记率≥95％，当前体分子与核素的投料摩尔比为3-50时，其标记率≥99％，此时我们认为标记反应已完全进行。
实施例4淬灭时间的控制
反应相溶液A的配制：在反应容器中依次加入10mCi无载体氯化镥[177Lu]溶液(约10μL)、10μL EB-FAPI前体溶液、60μL注射用水、20μL醋酸-醋酸钠缓冲盐溶液(其中含有50mg/mL龙胆酸)，将其混合均匀，该混合溶液为反应相溶液A。其中前体EB-FAPI为式Ⅳ的化合物，R＝H。
反应相溶液B的配制：在反应容器中加入10mCi无载体氯化镥[177Lu]溶液、20μL醋酸-醋酸钠缓冲盐溶液(其中含有50mg/mL龙胆酸)、60μL注射用水，混合均匀后，将此混合溶液在室温下静置3分钟(即淬灭时间)。然后继续向该反应容器中加入10μL EB-FAPI前体溶液，并混合均匀，该混合溶液为反应相溶液B。其中前体EB-FAPI为式Ⅳ的化合物，R＝H。
反应相溶液C的配制：除淬灭时间为15分钟外，其余与反应相溶液B相同。
加热反应与冷却：与处方(1)相同。
配辅与稀释：与处方(1)相同。
放射化学纯度检测：配辅稀释结束后，立即取150μL反应相溶液进行HPLC检测，计算放射化学纯度(HPLC)＝标记化合物的峰面积÷总峰面积。
反应相溶液A、B、C的放射化学纯度分别为93％、99％、99％。实验结果表明，在加入前体溶液之前，将含有核素溶液、缓冲盐与第一稳定剂的混合溶液静置一小段时间后(淬灭时间)再加入前体分子进行反应的投料顺序，能够明显提高API的初始放射化学纯度。因为经过一段淬灭时间，第一稳定剂与核素溶液充分接触，使溶液中由于高放射性而产生的大量自由基被第一稳定剂所淬灭，以减少后续加入前体分子时自由基对标记前体分子的破坏。淬灭时间取决于核素的种类与起始活度，笼统而言，淬灭时间控制在0.1-20分钟，优选3-10分钟。
实施例5[68Ga]Ga-EB-FAPI的标记
反应相溶液的配制：用0.1M盐酸淋洗市售锗镓发生器得到68Ga盐酸溶液，取68Ga盐酸溶液5mCi于反应容器中，加入20μL醋酸钠溶液(其中含有50mg/mL龙胆酸)、150μL注射用水，混合均匀后，将此混合溶液在室温下静置6分钟，然后向该反应容器中加入5μL EB-FAPI前体溶液， 加入注射用水使溶液总体积为0.3mL，并混合均匀，该混合溶液为反应相溶液。其中前体EB-FAPI为式Ⅳ的化合物，R＝H。
加热反应与冷却：将上述反应相溶液置于预热至95℃的加热器中反应30分钟，反应结束后取出反应容器冷却5分钟。
纯化：使用C18小柱对反应相溶液进行纯化，随后用0.4mL无水乙醇将标记络合物淋洗至产品瓶中。
配辅与稀释：向产品瓶中继续加入200μL配辅溶液，配辅溶液中含有45mg/mL龙胆酸、0.5mg/mL喷替酸和2.0mg/mL聚山梨酯80。最后向产品瓶中加入氯化钠注射液将总体积稀释至1.0mL，得到最终制剂溶液。
最终制剂溶液中含有9mg/mL龙胆酸、体积分数为40％的乙醇(即315.6mg/mL)、0.1mg/mL喷替酸、0.4mg/mL聚山梨酯80，其中API分子[177Lu]Lu-EB-FAPI在校准时刻的活度浓度为5mCi/mL，校准时刻是指生产结束时间(T0)。将制剂溶液储存于稳定性箱中，储存温度设置为25℃，储存湿度设置为60％RH。
在T0时刻使用Radio-HPLC检测制剂溶液的放射化学纯度为99％，在T0时刻使用ITLC检测制剂溶液的放射化学纯度为100％。
在T0+5h时刻取出稳定箱中的制剂溶液150μL，使用Radio-HPLC检测制剂溶液的放射化学纯度为93％，使用ITLC检测制剂溶液的放射化学纯度为100％。
虽然本案已以实施例揭露如上，然其并非用以限定本案，任何所属技术领域中具有通常知识者，在不脱离本案的精神和范围内，当可作些许的改动与润饰，故本案的保护范围当视后附的专利申请范围所界定者为准。

Claims (63)

  1. 放射性药物水溶液，其包括：如式Ⅰ所示的化合物或其药学上可接受的酯、酰胺、溶剂化物、盐、或如式Ⅰ所示的化合物或其药学上可接受的酯的盐、或如式Ⅰ所示的化合物或其药学上可接受的酰胺的盐、或如式Ⅰ所示的化合物或其药学上可接受的酯的溶剂化物、或如式Ⅰ所示的化合物或其药学上可接受的酰胺的溶剂化物、或如式Ⅰ所示的化合物或其药学上可接受的盐的溶剂化物，与放射性金属核素形成的络合物，以及以总浓度为0.5-400mg/mL存在的稳定剂，
    其中，
    L1是—(CH2)m—，其中m是0至12的整数，其中每个CH2可以单独地用—O—、—NH(CO)—、或—(CO)NH—替换，条件是没有两个相邻的CH2基团被替换；
    L2是C1-C60连接基团，任选地包括—O—、—S—、—S(O)—、—S(O)2—、—N(R)—、—C(＝O)—、—C(＝O)O—、—OC(＝O)—、—N(R)C(＝O)—、—C(＝O)N(R)—、—OC(＝O)O—、—N(R)C(＝O)O—、—OC(＝O)N(R)—、其中每个R为H或C1-C6烷基；
    L3是—(CH2)n—，其中n是0至12的整数，其中每个CH2可以单独地用—O—、—NH(CO)—、或—(CO)NH—替换，条件是没有两个相邻的CH2基团被替换；
    Ch是螯合基团；
    Tg是靶向基团。
  2. 根据权利要求1所述的药物水溶液，其中，L1为—NH(CO)—，L3为—NH(CO)CH2—。
  3. 根据权利要求1所述的药物水溶液，其中，Ch选自
  4. 根据权利要求3所述的药物水溶液，其中，Ch为
  5. 根据权利要求1所述的药物水溶液，其中，Tg选自能够靶向生长抑素受体(SSTR)、前列腺特异性膜抗原(PSMA)、成纤维细胞活化蛋白(FAP)、叶酸受体(FR)、表皮生长因子受体或整合素的化合基团。
  6. 根据权利要求5所述的药物水溶液，其中，Tg选自
  7. 根据权利要求1所述的药物水溶液，其中，式Ⅰ为式Ⅱ、式Ⅲ、式Ⅳ 或式Ⅴ，
  8. 根据权利要求1所述的药物水溶液，其中，所述放射性金属核素选自177Lu、99mTc、68Ga、64Cu、67Cu、111In、86Y、90Y、89Zr、186Re、188Re、153Sm、82Rb、166Ho、225Ac、212Pb、213Bi、212Bi或227Th。
  9. 根据权利要求1～8中任一项所述的药物水溶液，其中，所述稳定剂 选自龙胆酸及其盐、抗坏血酸及其盐、组氨酸、半胱氨酸及其盐、甲硫氨酸、硒甲硫氨酸、硫代硫酸盐、麦芽糖、肌醇、苯甲醇、海藻糖、聚维酮、烟酰胺、乙醇、姜黄素、褪黑素中的一种或两种以上。
  10. 根据权利要求9所述的药物水溶液，其中，所述稳定剂选自龙胆酸、乙醇、甲硫氨酸中的一种或两种以上。
  11. 根据权利要求10所述的药物水溶液，其中，在形成所述络合物的反应期间以及反应结束后分别加入所述稳定剂。
  12. 根据权利要求11所述的药物水溶液，其中，在形成所述络合物的反应期间加入的稳定剂选自龙胆酸及其盐、抗坏血酸及其盐、组氨酸、半胱氨酸及其盐、甲硫氨酸、硒甲硫氨酸、硫代硫酸盐、麦芽糖、肌醇、苯甲醇、海藻糖、聚维酮、烟酰胺、乙醇、姜黄素、褪黑素中的一种或两种以上；
    在形成所述络合物的反应结束后加入的稳定剂选自龙胆酸及其盐、抗坏血酸及其盐、组氨酸、半胱氨酸及其盐、甲硫氨酸、硒甲硫氨酸、硫代硫酸盐、麦芽糖、肌醇、苯甲醇、海藻糖、聚维酮、烟酰胺、乙醇、姜黄素、褪黑素中的一种或两种以上。
  13. 根据权利要求12所述的药物水溶液，其中，在形成所述络合物的反应期间加入的稳定剂为龙胆酸；在形成所述络合物的反应结束后加入的稳定剂为龙胆酸、乙醇或甲硫氨酸。
  14. 根据权利要求12所述的药物水溶液，其中，在形成所述络合物的反应期间和反应结束后加入的稳定剂均选自龙胆酸，且所述龙胆酸在所述药物水溶液中以0.1-10mg/mL的总浓度存在。
  15. 根据权利要求14所述的药物水溶液，其中，所述龙胆酸在所述药物水溶液中以0.5-5mg/mL的总浓度存。
  16. 根据权利要求12所述的药物水溶液，其中，在形成所述络合物的反应期间加入的稳定剂为龙胆酸，在形成所述络合物的反应结束后加入的稳定剂为乙醇，并且在所述药物水溶液中龙胆酸以0.1-10mg/mL的浓度存在，乙醇以0-400mg/mL的浓度存在。
  17. 根据权利要求16所述的药物水溶液，其中，在所述药物水溶液中龙胆酸以0.5-5mg/mL的浓度存在。
  18. 根据权利要求12所述的药物水溶液，其中，在形成所述络合物的 反应期间加入的稳定剂为龙胆酸，在形成所述络合物的反应结束后加入的稳定剂为甲硫氨酸，并且在所述药物水溶液中龙胆酸以0.1-10mg/mL的浓度存在，甲硫氨酸以0-50mg/mL的浓度存在。
  19. 根据权利要求18所述的药物水溶液，其中，在所述药物水溶液中龙胆酸以0.5-5mg/mL的浓度存在。
  20. 根据权利要求12的药物水溶液，其中还包括缓冲液，所述缓冲液可选自醋酸盐、柠檬酸盐、磷酸盐或甲酸盐溶液；所述缓冲液中的缓冲盐在所述药物水溶液中的浓度为0.005-0.5M。
  21. 根据权利要求12所述的药物水溶液，其中还包括助溶剂，所述助溶剂选自聚山梨酯20、聚山梨酯40、聚山梨酯60、聚山梨酯80、泊洛沙姆188、聚氧乙烯蓖麻油、司盘、乙醇、丙二醇、丙三醇、聚乙二醇(平均分子量为200～8000)、山梨醇、二甲基亚砜、十二烷基硫酸钠中的一种或两种以上。
  22. 根据权利要求21所述的药物水溶液，其中，所述助溶剂为聚山梨酯80。
  23. 根据权利要求12所述的药物水溶液，其中还包括游离核素螯合剂，所述游离核素螯合剂可选自喷替酸及其盐；所述游离核素螯合剂在所述药物水溶液中以0.005-0.1mg/mL的浓度存在。
  24. 根据权利要求1～23中任一项所述的药物水溶液，其特征在于，所述络合物在药物水溶液中以0.037-1850MBq/mL的活度浓度存在。
  25. 制备放射性药物水溶液的方法，所述放射性药物水溶液包含放射性核素与伊文思蓝衍生物分子形成的络合物，其特征在于，包括如下步骤：
    将含有第一稳定剂的溶液与含有放射性核素的溶液在反应容器中混合；
    给定时间后，将含有所述伊文思蓝衍生物分子的溶液加入至所述反应容器，所述给定时间为0.1分钟～20分钟
    所述伊文思蓝衍生物分子与放射性核素发生反应得到所述放射性核素络合物；
    反应给定时间后向所述反应容器中加入含有第二稳定剂的溶液；
    回收得到的所述放射性药物水溶液；
    其中，所述伊文思蓝衍生物分子为如式Ⅰ所示的化合物或其药学上可接 受的酯、酰胺、溶剂化物、盐、或如式Ⅰ所示的化合物或其药学上可接受的酯的盐、或如式Ⅰ所示的化合物或其药学上可接受的酰胺的盐、或如式Ⅰ所示的化合物或其药学上可接受的酯的溶剂化物、或如式Ⅰ所示的化合物或其药学上可接受的酰胺的溶剂化物、或如式Ⅰ所示的化合物或其药学上可接受的盐的溶剂化物，
    其中，
    L1是—(CH2)m—，其中m是0至12的整数，其中每个CH2可以单独地用—O—、—NH(CO)—、或—(CO)NH—替换，条件是没有两个相邻的CH2基团被替换；
    L2是C1-C60连接基团，任选地包括—O—、—S—、—S(O)—、—S(O)2—、—N(R)—、—C(＝O)—、—C(＝O)O—、—OC(＝O)—、—N(R)C(＝O)—、—C(＝O)N(R)—、—OC(＝O)O—、—N(R)C(＝O)O—、—OC(＝O)N(R)—、其中每个R为H或C1-C6烷基；
    L3是—(CH2)n—，其中n是0至12的整数，其中每个CH2可以单独地用—O—、—NH(CO)—、或—(CO)NH—替换，条件是没有两个相邻的CH2基团被替换；
    Ch是螯合基团；
    Tg是靶向基团。
  26. 根据权利要求25所述的方法，其特征在于，在给定时间后，将含有所述伊文思蓝衍生物分子的溶液加入至所述反应容器的步骤中，所述给定时间为3分钟～10分钟。
  27. 根据权利要求25所述的方法，其特征在于，所述放射性核素选自177Lu、99mTc、68Ga、64Cu、67Cu、111In、86Y、90Y、89Zr、186Re、188Re、153Sm、82Rb、166Ho、225Ac、212Pb、213Bi、212Bi或227Th。
  28. 根据权利要求25所述的方法，其特征在于，式Ⅰ中的Ch选自
  29. 根据权利要求25所述的方法，其特征在于，式Ⅰ中的Ch为
  30. 根据权利要求25所述的方法，其特征在于，式Ⅰ中的Tg选自能够靶向生长抑素受体(SSTR)、前列腺特异性膜抗原(PSMA)、成纤维细胞活化蛋白(FAP)、叶酸受体(FR)、表皮生长因子受体或整合素的化合基团。
  31. 根据权利要求25所述的方法，其特征在于，所述伊文思蓝衍生物分子选自选自如式Ⅱ、式Ⅲ、式Ⅳ或式Ⅴ所示的化合物，

  32. 根据权利要求25～31中任一项所述的方法，其中，所述含有放射性核素的溶液是从原料瓶中取出后加入至所述反应容器中，所述方法还包括：
    以冲洗液对所述原料瓶进行冲洗，并将冲洗后的溶液转入所述反应容器中与所述含有放射性核素的溶液混合。
  33. 根据权利要求32所述的方法，其特征在于，所述冲洗液为水溶液。
  34. 根据权利要求32所述的方法，其特征在于，所述冲洗液选自含有第一稳定剂的溶液、含有缓冲盐的溶液、水或氯化钠注射液。
  35. 根据权利要求32～34中任一项所述的方法，其特征在于，使用所述冲洗液重复冲洗一次或更多次。
  36. 根据权利要求25所述的方法，其特征在于，在所述伊文思蓝衍生 物分子与放射性核素发生反应的步骤中，所述伊文思蓝衍生物分子与放射性核素之间的摩尔比为1.5–50。
  37. 根据权利要求36所述的方法，其特征在于，所述伊文思蓝衍生物分子与放射性核素之间的摩尔比为5–20。
  38. 根据权利要求25所述的方法，其特征在于，在所述伊文思蓝衍生物分子与放射性核素发生反应的步骤中，反应温度为50-100℃，反应时间为5-60分钟。
  39. 根据权利要求38所述的方法，其特征在于，所述反应温度为60-80℃。
  40. 根据权利要求38所述的方法，其特征在于，所述反应时间为10-30分钟。
  41. 根据权利要求25所述的方法，其特征在于，所述第一稳定剂选自龙胆酸及其盐、抗坏血酸及其盐、组氨酸、半胱氨酸及其盐、甲硫氨酸、硒甲硫氨酸、硫代硫酸盐、麦芽糖、肌醇、苯甲醇、海藻糖、聚维酮、烟酰胺、乙醇、姜黄素、褪黑素中的一种或两种以上。
  42. 根据权利要求41所述的方法，其特征在于，所述第一稳定剂为龙胆酸。
  43. 根据权利要求41或42任一项所述的方法，其特征在于，在所述伊文思蓝衍生物分子与放射性核素发生反应的步骤中，在反应相溶液中第一稳定剂的浓度为0.6-20.0mg/mL。
  44. 根据权利要求25所述的方法，其特征在于，所述第二稳定剂选自龙胆酸及其盐、抗坏血酸及其盐、组氨酸、半胱氨酸及其盐、甲硫氨酸、硒甲硫氨酸、硫代硫酸盐、麦芽糖、肌醇、苯甲醇、海藻糖、聚维酮、烟酰胺、乙醇、姜黄素、褪黑素中的一种或两种以上。
  45. 根据权利要求44所述的方法，其特征在于，所述第二稳定剂选自龙胆酸、乙醇或甲硫氨酸。
  46. 根据权利要求44或45任一项所述的方法，其特征在于，在所述放射性药物水溶液中，所述第二稳定剂的浓度为0-400mg/mL。
  47. 根据权利要求25所述的方法，其特征在于，在所述伊文思蓝衍生物分子与放射性核素发生反应之前加入缓冲盐溶液。
  48. 根据权利要求47所述的方法，其特征在于，所述缓冲盐溶液存在 于所述含有第一稳定剂的溶液中。
  49. 根据权利要求47或48任一项所述的方法，其特征在于，所述缓冲盐溶液选自醋酸盐、柠檬酸盐、磷酸盐或甲酸盐溶液。
  50. 根据权利要求49所述的方法，其特征在于，所述缓冲盐溶液为醋酸-醋酸钠缓冲盐溶液。
  51. 根据权利要求25所述的方法，其特征在于，在反应给定时间后向所述反应容器中加入含有第二稳定剂的溶液的步骤还包括向所述反应容器中加入助溶剂。
  52. 根据权利要求51所述的方法，其特征在于，所述助溶剂选自聚山梨酯20、聚山梨酯40、聚山梨酯60、聚山梨酯80、泊洛沙姆188、聚氧乙烯蓖麻油、司盘、乙醇、丙二醇、丙三醇、聚乙二醇(平均分子量为200～8000)、山梨醇、二甲基亚砜、十二烷基硫酸钠中的一种或两种以上。
  53. 根据权利要求52所述的方法，其特征在于，所述助溶剂为聚山梨酯80。
  54. 根据权利要求25所述的方法，其特征在于，在反应给定时间后向所述反应容器中加入含有第二稳定剂的溶液的步骤还包括向所述反应容器中加入游离核素螯合剂，所述螯合剂可选自喷替酸及其盐。
  55. 根据权利要求54所述的方法，其特征在于，所述螯合剂为喷替酸。
  56. 根据权利要求25所述的方法，其特征在于，还包括使放射性药物水溶液通过0.22μm滤膜过滤除菌。
  57. 根据权利要求56所述的方法，其特征在于，于加入含有第二稳定剂的溶液后进行所述过滤除菌。
  58. 根据权利要求25所述的方法，其特征在于，还包括对所述放射性水溶液进行稀释。
  59. 根据权利要求58所述的方法，其特征在于，于加入含有第二稳定剂的溶液后，加入氯化钠注射液进行稀释。
  60. 放射性药物水溶液，其是由权利要求25～59中任一项所述的方法制备得到。
  61. 根据权利要求60所述的放射性药物水溶液，其中，所述放射性药物水溶液为权利要求1～24中任一项所述的放射性药物水溶液。
  62. 权利要求1～24中任一项所述的药物水溶液或权利要求25～59中任 一项所述的方法在肿瘤的放射治疗和/或诊断中的用途。
  63. 一种肿瘤的放射性治疗方法，其包括向有需要的受试者施用权利要求1～24中任一项所述的药物水溶液。
PCT/CN2023/101452 2022-04-20 2023-06-20 放射性伊文思蓝衍生物药物水溶液及其制备方法和用途 WO2023202730A2 (zh)

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