CN111362828A - A kind of18F-labeled fluoropropionylated ornithine as well as preparation method and application thereof - Google Patents
A kind of18F-labeled fluoropropionylated ornithine as well as preparation method and application thereof Download PDFInfo
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- C07C233/46—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by carboxyl groups with the substituted hydrocarbon radical bound to the nitrogen atom of the carboxamide group by an acyclic carbon atom
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Abstract
The invention provides a18F-labeled fluoropropionylated ornithine as well as a preparation method and application thereof belong to the technical field of radiopharmaceutical chemistry. The invention provides18F-labeled fluoropropionylated ornithine as N5‑(2‑[18F]-fluoropropionyl) ornithine. The invention provides18The F-labeled fluoropropionylated ornithine has good stability, higher uptake in tumor tissues and higher tumor/normal tissue ratio, and can be used as a PET-CT tumor imaging agent. In addition, the present invention provides the above18The preparation method of F-marked fluoropropionylated ornithine provided by the invention has the advantages of easily obtained raw materials and simple processThe method is simple, easy to operate and convenient for realizing automatic preparation.
Description
Technical Field
The invention relates to the technical field of radiopharmaceutical chemistry, in particular to a radioactive drug18F-marked fluoropropionylated ornithine as well as a preparation method and application thereof.
Background
Positron Emission Tomography (PET) technology is used as an important representative of functional imaging, is mature day by day in application of tumor diagnosis, staging and re-staging, curative effect monitoring and evaluation and radiotherapy target region delineation, and gradually develops to predictability and specificity. [18F]Fluorodeoxyglucose ([ alpha ], [ alpha18F]FDG) is a PET imaging agent for tumor metabolism which is most clinically applied, but due to the universality of metabolism, high uptake can also exist in non-tumor tissues such as brain, inflammation, tumor-like lesions and the like, and false negative results are easily caused in the diagnosis of tumors which do not cause obvious glucose metabolism abnormality such as mucinous adenocarcinoma, bronchoalveolar carcinoma, prostate cancer and the like. Therefore, the research and development of the [ 2 ]18F]FDG has a higher sensitivity, a better specificity for tumors.
Polyamines are substances that are necessary for maintaining vital activities such as cell growth and differentiation, as metabolic control substances that are widely distributed in the living body. Under normal physiological conditions, polyamines are regulated by a series of mechanisms of synthesis, catabolism and transport across membranes, thereby maintaining the intracellular polyamine content in a state of homeostasis. Studies have demonstrated that tumor development is associated with an imbalance in polyamine metabolism, and rapid proliferation of tumor cells is dependent on high levels of intracellular polyamines, which in turn upregulate polyamine synthase activity. Ornithine is used as a main raw material for in vivo polyamine synthesis, and is decarboxylated under the action of Ornithine Decarboxylase (ODC) to synthesize polyamines. ODC is the first-order rate-limiting enzyme regulating polyamine biosynthesis in cells and plays an important role in regulating polyamine levels in cells. Research shows that the overexpression of oncogenes can induce a large amount of ODC generation, so that intracellular polyamine levels are increased, and the ODC activity is remarkably increased in glioma, gastric cancer, prostatic cancer, melanoma, breast cancer, colorectal cancer and the like. Changes in the intracellular expression levels of ODCs can also affect various growth factor functions, such as stimulating oncogene expression, etc. In addition, the activity of ODC begins to increase early in the process of cell stimulation, which is earlier than the change of DNA synthesis, so that ODC is considered as a biological factor for making "early" response, and has become a target for anti-tumor therapy.
Considering that ornithine is used as a main material for polyamine synthesis, and the synthesis of polyamine is completed by decarboxylation under the action of ODC, a series of single-photon nuclides are reported in a patent (patent number: PCT/SG2011/000160)99mTc-labeled ornithine and derivatives thereof are useful as tumor imaging agents in Single Photon Emission Computed Tomography (SPECT) techniques. Because PET has better resolution than SPECT, the positron nuclide labeled ornithine compound is developed to be applied to nuclear medicine tumor image diagnosis, and the positron nuclide labeled ornithine compound has important significance for early diagnosis and curative effect monitoring of malignant tumors.
Disclosure of Invention
The invention aims to provide18F-marked fluoropropionylated ornithine as well as a preparation method and application thereof. The invention provides18The F-labeled fluoropropionylated ornithine has good stability, high tumor uptake and good retention, has a high tumor/normal tissue ratio, and can be used as a PET-CT tumor imaging agent.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a18F-labeled fluoropropionylated ornithine as N5-(2-[18F]-fluoropropionyl) ornithine having the structure of formula I:
the invention provides the technical scheme18A method for preparing F-labeled fluoropropionylated ornithine, comprising the steps of:
(1) will N5- (2-bromopropane)Acyl) -N2-Boc-ornithine tert-butyl ester, solvent and [ 2 ]18F]F-Mixing the sources, carrying out nucleophilic reaction to obtain N5-(2-18F-fluoropropionyl) -N2-Boc-ornithine tert-butyl ester; the [ 2 ]18F]F-The source comprises18F]F-And a catalyst;
(2) the N is5-(2-18F-fluoropropionyl) -N2-Boc-ornithine tert-butyl ester is mixed with hydrochloric acid for hydrolysis reaction to obtain18F-labeled fluoropropionylated ornithine.
Preferably, the catalyst comprises a phase transfer catalyst and a base catalyst.
Preferably, the phase transfer catalyst is K222, 18-crown-6 or tetra-n-butyl ammonium bicarbonate, and the base catalyst is K2CO3、KHCO3、K2(COO)2Or Cs2CO3。
Preferably, the temperature of the nucleophilic reaction is 70-130 ℃, and the time is 5-40 min.
Preferably, the nucleophilic reaction is followed by liquid chromatography purification.
Preferably, the temperature of the hydrolysis reaction is 60-140 ℃ and the time is 5-15 min.
Preferably, the hydrolysis reaction further comprises: using NaOH solution or NaHCO3Neutralizing the system obtained after hydrolysis reaction with the solution, filtering with sterile filter membrane, and obtaining the product in the solution18F-labeled fluoropropionylated ornithine.
The invention provides the technical scheme18The F-labeled fluoropropionylated ornithine is applied to the preparation of a PET-CT tumor imaging agent.
Preferably, the tumor imaged by the PET-CT tumor imaging agent is prostate cancer, liver cancer, lung cancer, gastric cancer, glioma, colon cancer, bone fibrosarcoma, melanoma or breast cancer.
The invention provides a18F-labeled fluoropropionylated ornithine as N5-(2-[18F]-fluoropropionyl) ornithine. The invention provides18F-labelled fluorineThe alanylated ornithine has good stability, higher uptake in tumor tissues and higher tumor/normal tissue ratio (namely target/non-target ratio), and can be used as a PET-CT tumor imaging agent.
In addition, the present invention provides the above18The preparation method of F-labeled fluoropropionylated ornithine provided by the invention has the advantages of easily obtained raw materials, simple process, easy operation and convenience for realizing automatic preparation.
Drawings
FIG. 1 is N5-(2-[19F]-fluoropropionyl) -N2-HPLC uv chromatogram of Boc-ornithine tert-butyl ester;
FIG. 2 is N5-(2-[18F]-fluoropropionyl) ornithine from a sample;
FIG. 3 is N5-(2-[18F]-fluoropropionyl) ornithine;
FIG. 4 is N5-(2-18F-fluoropropionyl) -N2-preparative HPLC separation chromatogram of Boc-ornithine tert-butyl ester (A: HPLC UV chromatogram, B: HPLC radiochromatogram);
FIG. 5 is a PET-CT image obtained in application example 2;
FIG. 6 is a PET-CT image obtained in application example 3;
FIG. 7 is a PET-CT image obtained in application example 4.
Detailed Description
The invention provides a18F-labeled fluoropropionylated ornithine as N5-(2-[18F]-fluoropropionyl) ornithine having the structure of formula I:
the invention provides the technical scheme18A method for preparing F-labeled fluoropropionylated ornithine, comprising the steps of:
(1) will N5- (2-bromopropionyl) -N2-Boc-ornithine tert-butyl ester, solvent and [ 2 ]18F]F-The mixing of the sources is carried out,nucleophilic reaction to obtain N5-(2-18F-fluoropropionyl) -N2-Boc-ornithine tert-butyl ester; the [ 2 ]18F]F-The source comprises18F]F-And a catalyst;
(2) the N is5-(2-18F-fluoropropionyl) -N2-Boc-ornithine tert-butyl ester is mixed with hydrochloric acid for hydrolysis reaction to obtain18F-labeled fluoropropionylated ornithine.
In the present invention, unless otherwise specified, the starting materials and reagents used are commercially available products well known to those skilled in the art.
The invention converts N into5- (2-bromopropionyl) -N2-Boc-ornithine tert-butyl ester, solvent and [ 2 ]18F]F-Mixing the sources, carrying out nucleophilic reaction to obtain N5-(2-18F-fluoropropionyl) -N2-Boc-ornithine tert-butyl ester; the [ 2 ]18F]F-The source comprises18F]F-And a catalyst. The invention is directed to said N5- (2-bromopropionyl) -N2The source of Boc-ornithine tert-butyl ester is not particularly limited and may be made by oneself or purchased; in the embodiment of the invention, N is5- (2-bromopropionyl) -N2-Boc-ornithine tert-butyl ester is preferably prepared by the following steps:
will N2mixing-Boc-L-ornithine tert-butyl ester, 2-bromopropionyl bromide, triethylamine and a solvent, and carrying out an acylation reaction to obtain N5- (2-bromopropionyl) -N2-Boc-ornithine tert-butyl ester.
In the present invention, said N2The mol ratio of the-Boc-L-ornithine tert-butyl ester to the 2-bromopropionyl bromide to the triethylamine is preferably 1: 0.5-2: 1-3, and more preferably 1: 0.5-1: 2; triethylamine can neutralize HBr produced by the acylation reaction. In the present invention, the solvent for acylation reaction is preferably dichloromethane, more preferably anhydrous dichloromethane; the dosage of the solvent is not specially limited, and the acylation reaction is ensured to be smoothly carried out; in the embodiment of the present invention, N is2The ratio of the amount of-Boc-L-ornithine tert-butyl ester to the solvent is preferably 1g:20 mL.
In bookIn the invention, said N2The mixing process of the-Boc-L-ornithine tert-butyl ester, the 2-bromopropionyl bromide, the triethylamine and the solvent is preferably carried out at the temperature of-10 ℃, more preferably at the temperature of-5 ℃, and further preferably at the temperature of 0 ℃; in the embodiment of the invention, the four are mixed under the ice-water bath condition. In the present invention, said N2The preferred mode of mixing the tert-butyl-Boc-L-ornithine, 2-bromopropionyl bromide, triethylamine and the solvent is to mix N2Mixing the-Boc-L-ornithine tert-butyl ester and triethylamine with a solvent, uniformly stirring, then dropwise adding 2-bromopropionyl bromide into the obtained mixture, and preferably continuously stirring for 15-60 min, more preferably 30-35 min after dropwise adding is finished so as to uniformly mix the components; the invention adopts a dropwise adding mode to add the 2-bromopropionyl bromide, which can avoid a large amount of heat in the system; the dropping rate is not particularly limited in the present invention, and may be a dropping rate well known to those skilled in the art.
Will N2After the tert-butyl-Boc-L-ornithine, the 2-bromopropionyl bromide, the triethylamine and the solvent are mixed, the temperature of the system is preferably raised to the temperature of the acylation reaction for carrying out the acylation reaction. In the present invention, the temperature of the acylation reaction is preferably room temperature, i.e., no additional heating or cooling is required; in the examples of the present invention, the room temperature is specifically 23 ℃. The temperature rising rate is not specially limited, and the temperature can reach the room temperature; in the embodiment of the present invention, the ice-water bath is removed, and the obtained reaction solution is naturally warmed up to room temperature.
In the invention, the time of the acylation reaction is preferably 0.1-24 h, and more preferably 18-24 h; the time of the acylation reaction is preferably from when the reaction solution is warmed to room temperature. According to the invention, TLC (thin layer chromatography) is preferably adopted to verify whether the acylation reaction is complete, and if the reaction is complete, the subsequent steps are carried out, otherwise, the reaction is continued.
After the acylation reaction is completed, the material obtained in the acylation reaction is preferably filtered, the obtained filtrate is extracted by ethyl acetate and water, the organic phases are combined, and anhydrous Na is utilized2SO4Drying and evaporating the solvent to obtain N5- (2-bromopropionyl) -N2-Boc-ornithine tert-butyl ester; what is needed isThe filtration enables the removal of insoluble materials from the mass obtained from the acylation reaction. The extraction times and the amounts of ethyl acetate and water used in the present invention are not particularly limited, and the technical solutions well known to those skilled in the art can be adopted.
To obtain N5- (2-bromopropionyl) -N2After Boc-ornithine tert-butyl ester, N is introduced into the reaction mixture5- (2-bromopropionyl) -N2-Boc-ornithine tert-butyl ester, solvent and [ 2 ]18F]F-Mixing the sources, carrying out nucleophilic reaction to obtain N5-(2-18F-fluoropropionyl) -N2-Boc-ornithine tert-butyl ester. In the present invention, the solvent for nucleophilic reaction is preferably acetonitrile, dimethyl sulfoxide, N-dimethylformamide or tetrahydrofuran, more preferably acetonitrile; the dosage of the solvent is not specially limited, and the nucleophilic reaction is ensured to be smoothly carried out5- (2-bromopropionyl) -N2The dosage ratio of the Boc-ornithine tert-butyl ester to the solvent is preferably 1-20 mg: 0.5-2.0 mL, and more preferably 10-15 mg: 0.8-1.5 mL.
In the present invention, the term18F]F-The source comprises18F]F-And a catalyst; the catalyst preferably comprises a phase transfer catalyst, preferably K222 (i.e. Kryptofix2.2.2), 18-crown-6 or tetra-n-butylammonium bicarbonate (TBAHCO)3) The base catalyst is preferably K2CO3、KHCO3、K2(COO)2Or Cs2CO3。
The present invention is directed to the [ 2 ]18F]F-The method for producing the source is not particularly limited, and those conventional in the art are used18F]F-The method of producing a source, in the present embodiment, the [ alpha ], [ beta ], [18F]F-The method of preparation of the source preferably comprises the steps of:
capture [ 2 ] with Sep Pak QMA column18F]F-Then, eluting by using an eluant to obtain an eluent; removing the solvent in the eluent, mixing the remainder with acetonitrile, and removing the solvent in the mixed material liquid, wherein the remainder is the [ alpha ], [ beta ] -isomer, or [ beta18F]F-A source.
In the present invention, the term18F]F-Preferably a value produced by a nuclear reaction using a cyclotron18F]HF. The present invention captures the protein using Sep Pak QMA column18F]F-The method (2) is not particularly limited, and a method known to those skilled in the art may be used. In the invention, the dosage of the eluent is preferably 0.5-3 mL, and more preferably 1.5 mL; the eluent contains a phase transfer catalyst and an alkali catalyst, the mass ratio of the phase transfer catalyst to the alkali catalyst is preferably 17.7:4.1, and the concentration of the phase transfer catalyst in the eluent is preferably 11-12 mg/mL; the solvent of the eluent is preferably an acetonitrile-water mixed solvent, wherein the volume ratio of acetonitrile to water is preferably 10: 1. In the present example, the obtained [ alpha ], [18F]F-The source is preferably marked with 1-25 mg of N5- (2-bromopropionyl) -N2-Boc-ornithine tert-butyl ester, more preferably 10 to 13 mg. The method for removing the solvent in the present invention is not particularly limited, and the solvent may be evaporated by heating.
In the present invention, said N5- (2-bromopropionyl) -N2-Boc-ornithine tert-butyl ester, solvent and [ 2 ]18F]F-The source is preferably mixed in such a way that N is5- (2-bromopropionyl) -N2-Boc-ornithine tert-butyl ester is dissolved in a solvent and then reacted with [ [ solution ] ] [ ]18F]F-And (4) mixing the sources.
In the invention, the temperature of the nucleophilic reaction is preferably 70-130 ℃, and more preferably 90-110 ℃; the time is preferably 5 to 50min, and more preferably 20 to 30 min. In the present invention, the nucleophilic reaction is preferably performed under a sealing condition.
After the nucleophilic reaction is completed, the reaction solution obtained by the nucleophilic reaction is preferably subjected to liquid chromatography purification in the invention. In the present invention, the liquid chromatography purification can remove the phase transfer catalyst from the reaction solution and can also completely remove the unreacted N5- (2-bromopropionyl) -N2In the present invention, the operating conditions for the liquid chromatography purification preferably include a semi-preparative C18 reverse phase column (5 μm, 250 × 10mm)The mobile phase is acetonitrile and water, the flow rate of the mobile phase is 3mL/min, gradient elution is adopted, and the conditions of the gradient elution are as follows: keeping the water for 0-1 min, and keeping the water for 90%; reducing the water content from 90% to 20% in 1-25 min; keeping the water for 20% in 25-35 min; collecting the eluent with the retention time of 17-19 min.
In the present invention, the eluate obtained by collection contains the product N5-(2-18F-fluoropropionyl) -N2-Boc-ornithine tert-butyl ester; removing the solvent in the eluent to obtain N5-(2-18F-fluoropropionyl) -N2-Boc-ornithine tert-butyl ester; the method for removing the solvent in the present invention is not particularly limited, and the solvent may be evaporated by heating.
To obtain N5-(2-18F-fluoropropionyl) -N2After Boc-ornithine tert-butyl ester, the invention provides said N5-(2-18F-fluoropropionyl) -N2-Boc-ornithine tert-butyl ester is mixed with hydrochloric acid for hydrolysis reaction to obtain18F-labeled fluoropropionylated ornithine. In the invention, the concentration of the hydrochloric acid is preferably 2-5 mol/L, and more preferably 4 mol/L; in the present invention, said N5- (2-bromopropionyl) -N2The dosage ratio of the Boc-ornithine tert-butyl ester to the hydrochloric acid is preferably 13.0mg: 0.2-1 mL.
In the invention, the temperature of the hydrolysis reaction is preferably 60-140 ℃, more preferably 105-115 ℃, and the time is preferably 5-15 min, more preferably 10 min.
In the present invention, the hydrolysis reaction preferably further comprises: using NaOH solution or NaHCO3Neutralizing the system obtained after hydrolysis reaction with the solution, filtering with sterile filter membrane, and obtaining the product in the solution18F-labeled fluoropropionylated ornithine. In the present invention, the NaOH solution or NaHCO3The concentration of the solution is preferably 0.2-6 mol/L, and more preferably 4 mol/L. In the present invention, NaOH solution or NaHCO is added3The solution has the function of neutralizing hydrochloric acid in a system obtained after hydrolysis reaction to obtain a solution with a proper pH value (the pH value is preferably 6.0-7.0), and the solution contains18F-labeled fluoropropionylated ornithine (referred to as18F-labeled fluoropropionylated ornithine injection), may be usedCan be directly used as a PET-CT imaging agent. The method of filtration with the sterile filtration membrane is not particularly limited in the present invention, and a method known to those skilled in the art may be used.
In the present invention, the18F-labeled fluoropropionylated ornithine injection18The radiochemical yield of F-labeled fluorobromopropionylated ornithine is 5-8%. In addition, the raw materials used in the preparation method provided by the invention are easy to obtain, the process is simple, the operation is easy, and the automatic preparation can be realized by using a conventional multifunctional synthesis mode.
The invention provides the technical scheme18The F-labeled fluoropropionylated ornithine is applied to the preparation of a PET-CT tumor imaging agent. In the invention, the preparation method of the technical scheme obtains18The F-labeled fluoropropionylated ornithine injection can be directly used as a PET-CT tumor imaging agent. In the present invention, the tumor imaged by the PET-CT tumor imaging agent is preferably any one of prostate cancer, liver cancer, lung cancer, stomach cancer, glioma, colon cancer, bone fibrosarcoma, melanoma and breast cancer.
The technical solution of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. It is to be understood that the described embodiments are merely exemplary of the invention, and not restrictive of the full scope of the invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1
Preparation of N5-(2-[18F]-fluoropropionyl) ornithine, wherein the reaction formulae of the acylation reaction, the nucleophilic reaction and the hydrolysis reaction are represented by the formulae a, b and c, respectively:
will N2-Boc-L-ornithine tert-butyl ester (1g, 3.473mmol), triethylamine (6.946mmol) and anhydrous dichloromethane (20mL) were mixed, stirred well in an ice water bath (0 ℃ C.), and slowly added dropwiseAdding 2-bromopropionyl bromide (0.25mL, 2.315mmol) into the reaction system, stirring thoroughly for 30min, returning to room temperature (23 deg.C), continuing stirring for acylation reaction for 20h, detecting by TLC that the reaction is finished, filtering, adding 50mL water into the obtained filtrate, extracting with ethyl acetate (3 × 50mL), combining organic phases, anhydrous Na2SO4Drying and evaporating the solvent to dryness to obtain 0.51g of a yellow powdery solid, i.e. N5- (2-bromopropionyl) -N2-Boc-ornithine tert-butyl ester; determination of N by high resolution Mass Spectrometry5- (2-bromopropionyl) -N2-Boc-ornithine tert-butyl ester has a molecular weight of [ M + Na]445.12, theoretical molecular weight 422.14;
capture [ 2 ] with Sep Pak QMA column18F]F-(produced from a cyclotron by a nuclear reaction18F]HF), elution is carried out using 1.5mL of an eluent containing 17.7mg of phase transfer catalyst K222 and 4.1mg of base catalyst K2CO3The solvent of the eluent is acetonitrile-water mixed solvent, and the volume ratio of acetonitrile to water is 10: 1; heating the eluate to 110 deg.C, removing solvent by azeotropic evaporation, adding 1mL acetonitrile, and evaporating solvent again to obtain the final product18F]F-A source;
the activity of which is 37MBq18F]F-Source and 1mLN5- (2-bromopropionyl) -N2-Boc-ornithine tert-butyl ester in acetonitrile (containing N)5- (2-bromopropionyl) -N213.0mg of-Boc-ornithine tert-butyl ester), sealing and heating to 100 ℃ for nucleophilic reaction for 30min, after the reaction is finished, purifying by HPLC, wherein a chromatographic column used for purification is a semi-preparative C18 reverse phase column (the specification is 5 mu m and 250 × 10mm), a mobile phase is acetonitrile and water, the flow rate of the mobile phase is 3mL/min, gradient elution is adopted for purification, the conditions of gradient elution are that water is kept for 90% for 0-1 min, water is reduced from 90% to 20% for 1-25 min, water is kept for 25-35 min and 20%, eluent with the retention time of 17-19 min is collected, and the solvent is evaporated to obtain N5-(2-18F-fluoropropionyl) -N2-Boc-ornithine tert-butyl ester;
at 100 ℃ to the N5-(2-18F-fluoropropionyl) -N2-BocAdding 0.2mL of hydrochloric acid with the concentration of 4mol/L into tert-butyl ornithine for hydrolysis reaction for 10min, then adding a NaOH solution with the concentration of 4mol/L for neutralization, and filtering the obtained mixed solution by using a sterile filter membrane to obtain N5-(2-[18F]-fluoropropionyl) ornithine injection, the pH value of which is 6.0-7.0, and the N is suitable for carrying out PET-CT imaging on a mouse with tumor, and normal saline is used for carrying out PET-CT imaging on the N5-(2-[18F]-fluoropropionyl) ornithine injection was adjusted to an activity concentration of 25 MBq/mL.
Will be corrected N5-(2-[18F]The activity of the (F) -fluoropropionyl) ornithine injection solution is divided by the value of the activity of the [ 2 ], [ solution of ] used18F]F-The activity of the source gave a corrected radiochemical yield of about 5% in this example.
Determination of N by HPLC5-(2-[18F]-fluoropropionyl) chemical purity and radiochemical purity of ornithine injection. FIG. 1 is N5-(2-19F-fluoropropionyl) -N2HPLC ultraviolet chromatogram of-Boc-ornithine tert-butyl ester, retention time corresponding to ultraviolet peak of 17.5min, FIG. 2 is N5-(2-[18F]-fluoropropionyl) ornithine with a retention time of 2.5min, and further determining radiochemical purity by Thin Layer Chromatography (TLC) using silica gel plate (1cm × 20cm) and developing solvent of 85% acetonitrile aqueous solution, the determination result is shown in FIG. 3, and determined by TLC, N5-(2-[18F]The Rf value of the-fluoropropionyl) ornithine injection solution is 0.42 (under the TLC condition [ [ solution ] ] [, ]18F]F-Rf value of less than 0.1), N is seen5-(2-[18F]-fluoropropionyl) ornithine injection having a radiochemical purity of greater than 99%. FIG. 4 is N5-(2-18F-fluoropropionyl) -N2-Boc-ornithine tert-butyl ester preparative HPLC separation chromatogram, wherein a is an HPLC uv chromatogram and B is an HPLC radioactive chromatogram. As can be seen from FIGS. 1 to 4, intermediate N5-(2-18F-fluoropropionyl) -N2The emission peak of-Boc-ornithine tert-butyl ester is 17-19 min, and N5-(2-19F-fluoropropionyl) -N2Retention of UV peak of-Boc-Ornithine tert-butyl esterMeta-proximity, indicating that the product obtained after nucleophilic reaction by purification is N5-(2-18F-fluoropropionyl) -N2-Boc-ornithine tert-butyl ester.
Will N5-(2-[18F]-fluoropropionyl) ornithine injection was left at room temperature, and the N was added using physiological saline5-(2-[18F]-fluoropropionyl) ornithine injection, the activity concentration of which is adjusted to 25MBq/mL, and the radiochemical purity of which is measured by HPLC at 0min, 30min, 60min and 120min after the adjustment of the activity concentration, the results show that the radiochemical purity is respectively 98%, 97% and 96% at 30min, 60min, 90min and 120min, and are all higher than 95%, which indicates that N is higher than 95%5-(2-[18F]-fluoropropionyl) ornithine has excellent in vitro stability.
Application example 1
Abnormal toxicity test:
Mice tail vein injected with decayed N5-(2-[18F]-fluoropropionyl) ornithine injection has no adverse reaction and death phenomenon after 48 hours and one week of observation, and no organ injury is observed after the observation after the dissection.
The above results illustrate that N5-(2-[18F]-fluoropropionyl) ornithine injection has no toxicity to organisms and can be further used for in vivo research.
Application example 2
PET-CT imaging test:
3 DU-145 nude mice with prostate cancer were fixed in a Minerve mouse animal chamber (Minerve) with a constant temperature of 37 ℃ after being anesthetized by 2% isoflurane gas induction
Verterinaire, Esternay, France) to keep the animal's body temperature constant while the animals were given 2L/min air and 2% isoflurane mixed gas inhalation anaesthesia and then developed by tail vein injectionImaging agent (i.e., N obtained in example 1)5-(2-[18F]-fluoropropionyl) ornithine injection, 0.2mL, 5MBq), while performing PET static scan and CT scan for 10min after 30min, 60min, 90min and 120min of injection of imaging agent, respectively, and performing image reconstruction and quantitative analysis of each tissue of interest, determining the percentage injection dose rate per gram of tissue (% ID/g), and calculating the uptake ratio of tumor to normal tissue (T/NT); the resulting PET-CT image is shown in FIG. 5.
As can be seen from FIG. 5, in the case of injecting the developer (N)5-(2-[18F]-fluoropropionyl) ornithine injection) has obvious imaging agent uptake at tumor sites and good retention, and at 60min, the relative uptake ratio of tumor/muscle is about 3.1, and the contrast with surrounding tissues is obvious. Except for kidney and pancreas, the uptake of imaging agents was lower in normal tissues or organs than in tumor tissues.
Application example 3
PET-CT imaging test:
3 mice with lotus S180 (bone fibrosarcoma) are fixed in a Minerve mouse animal chamber (Minerve) with constant temperature of 37 ℃ after being induced and anesthetized by isoflurane gas with 2 percent
In Veertilaire, Esternay, France) to keep the animal's body temperature constant, the animals were given 2L/min air and 2% isoflurane mixed gas for inhalation anesthesia and then injected via tail vein (i.e. N obtained in example 1)5-(2-[18F]-fluoropropionyl) ornithine injection, 0.2mL, 5MBq), while PET static and CT scans were performed for 10min 60min after injection of imaging agent, and the uptake ratio (T/NT) of tumor to normal tissue was calculated; the resulting PET-CT image is shown in FIG. 6.
As can be seen from FIG. 6, in the case of injecting the developer (N)5-(2-[18F]-fluoropropionyl) ornithine injection) 60min, the contrast agent uptake at the tumor site was significant, the relative tumor/muscle uptake ratio was about 2.5, and a significant differentiation of tumor from surrounding tissues was observed.
Application example 4
PET-CT imaging test:
3 gliomas were anesthetized by 2% isoflurane gas induction and fixed in a Minerve rat animal chamber (Minerve) with a constant temperature of 37 deg.C
In Veertilaire, Esternay, France) to keep the body temperature of the animals constant, the animals were given 3L/min of air mixed with 2% isoflurane for inhalation anesthesia and then injected via tail vein (i.e. N obtained in example 1)5-(2-[18F]-fluoropropionyl) ornithine injection, 0.3mL, 7.5MBq), while PET static and CT scans were performed for 10min 60min after injection of imaging agent and the uptake ratio (T/NT) of tumor to normal tissue was calculated; the resulting PET-CT image is shown in FIG. 7.
As can be seen from FIG. 7, in the case of injecting the developer (N)5-(2-[18F]-fluoropropionyl) ornithine injection) has a relatively clear boundary at the tumor site, and the relative uptake ratio of tumor/normal brain tissue is about 2.3, which is clearly distinguished from the surrounding tissue.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (10)
1. A kind of18F-labeled fluoropropionylated ornithine characterized by being N5-(2-[18F]-fluoropropionyl) ornithine having the structure of formula I:
2. the method of claim 118The preparation method of F-labeled fluoropropionylated ornithine is characterized by comprising the following steps of:
(1) will N5- (2-bromopropionyl) -N2-Boc-ornithine tert-butyl ester, solvent and [ 2 ]18F]F-Mixing the sources, carrying out nucleophilic reaction to obtain N5-(2-18F-fluoropropionyl) -N2-Boc-ornithine tert-butyl ester; the [ 2 ]18F]F-The source comprises18F]F-And a catalyst;
(2) the N is5-(2-18F-fluoropropionyl) -N2-Boc-ornithine tert-butyl ester is mixed with hydrochloric acid for hydrolysis reaction to obtain18F-labeled fluoropropionylated ornithine.
3. The method of claim 2, wherein the catalyst comprises a phase transfer catalyst and a base catalyst.
4. The process according to claim 2 or 3, wherein the phase transfer catalyst is K222, 18-crown-6 or tetra-n-butylammonium hydrogen carbonate and the base catalyst is K2CO3、KHCO3、K2(COO)2Or Cs2CO3。
5. The preparation method according to claim 2, wherein the temperature of the nucleophilic reaction is 70-130 ℃ and the time is 5-40 min.
6. The method according to claim 2 or 5, wherein the nucleophilic reaction is followed by liquid chromatography purification.
7. The method according to claim 2, wherein the hydrolysis reaction is carried out at a temperature of 60 to 140 ℃ for 5 to 15 min.
8. The method according to claim 2 or 7, further comprising, after the hydrolysis reaction: using NaOH solution or NaHCO3Neutralizing the system obtained after hydrolysis reaction with the solution, filtering with sterile filter membrane, and dissolving in the solutionTo obtain18F-labeled fluoropropionylated ornithine.
9. The method of claim 118The F-labeled fluoropropionylated ornithine is applied to the preparation of a PET-CT tumor imaging agent.
10. The use of claim 9, wherein the tumor imaged by the PET-CT tumor imaging agent is prostate cancer, liver cancer, lung cancer, gastric cancer, glioma, colon cancer, fibrosarcoma of bone, melanoma, or breast cancer.
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