CN114989166A - Tumor KRAS G12C mutation targeted positron tracer, preparation method and application - Google Patents
Tumor KRAS G12C mutation targeted positron tracer, preparation method and application Download PDFInfo
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Abstract
The invention relates to a tumor KRAS G12C mutation targeted positron tracer, a preparation method and application thereof. The existing KRAS mutation detection means can not completely meet the target requirement of accurate diagnosis and treatment. The invention provides a KRAS G12C mutant targeting compound for tumors 18 F/ 19 F-QZLO of 18 The positron tracer of F-QZLO is used for nuclide diagnosis of KRAS G12C mutation positive tumors, 19 F-QZLO was used as a stable standard. The invention labels the radioactive nuclide to AMG510 through glycol or polyethylene glycol chain 18 F, constructing a PET molecular imaging probe, using the PET molecular imaging probe as a tumor KRAS G12C mutation targeted positron tracer, and providing a stable standard substance 19 F-QZLO, applianceHas proper physicochemical and radiological properties and ideal biological characteristics, can be used for PET imaging of KRAS G12C mutation tumors, and is helpful for specific diagnosis of the tumors, selection of targeted therapeutic drugs and evaluation of curative effect.
Description
Technical Field
The invention relates to a Positron tracer of Positron Emission Computed Tomography (PET), in particular to a tumor murine sarcoma virus oncogene (KRAS) G12C mutation targeted Positron tracer, a preparation method and application.
Background
Lung cancer is a serious disease seriously threatening human health, and the incidence and the mortality of the lung cancer are the first of all tumors. In recent years, molecular targeted drugs represented by Epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKIs) have revolutionized the efficacy of EGFR mutant lung cancer, and the Progression-free survival (PFS) has been greatly increased from 4.6 to 6.7 months of conventional chemoradiotherapy to 8.4 to 13.1 months. However, clinical studies have also shown that 29% to 63% of EGFR mutant patients do not respond to EGFR-TKI treatment. Therefore, the accurate evaluation and prediction of the treatment effect of the EGFR-TKI of the lung cancer are difficult to realize by simply depending on the screening of EGFR mutation, and a new strategy for predicting the curative effect of the EGFR-TKI is urgently needed to be established.
KRAS mutation is one of the most common gene mutations in lung cancer, and KRAS mutation can remarkably reduce or even completely eliminate the response of lung cancer patients to EGFR-TKI treatment and is an important curative effect prediction marker. Based on the important role of KRAS mutation on lung cancer molecular targeted therapy decision and prognosis, the KRAS mutation screening is recommended to be classified as a Clinical molecular examination accepted by lung cancer patients routinely by a plurality of international organizations such as American Society of Clinical Oncology (ASCO); the National Comprehensive Cancer Network (NCCN) guidelines indicate that "whether KRAS mutation determines whether a patient also needs to detect other molecular therapeutic targets". Therefore, accurate detection of KRAS mutation is a great clinical requirement for accurate diagnosis and treatment of lung cancer. However, in the conventional KRAS mutation detection means in clinic, invasive needle biopsy is limited by the limitation of a puncture part and cannot accurately reflect the heterogeneity of KRAS, and circulating tumor DNA detection has the outstanding problem of too low sensitivity, and both cannot completely meet the target of accurate diagnosis and treatment. The molecular image can carry out real-time, quantitative and visual visualization on the living biological molecules.
In recent years, KRAS mutation targeting molecules AMG510 and ARS1620, which have been approved by FDA and marketed, can inhibit activity of KRAS G12C protein by irreversibly binding to the protein, and provide a new development opportunity for molecular imaging of KRAS mutation. Objective Response Rate (ORR, the proportion of patients with tumor volume reduction ≧ 30%) for treatment with 960mg AMG510 taken once daily orally achieved disease control (proportion of patients achieving complete remission, partial remission, and stable disease for more than 3 months) in 36% (95% CI:28-45), 81% (95% CI: 73-87). The Duration of median remission (DoR) was 10 months.
Disclosure of Invention
The invention aims to provide a KRAS G12C mutation targeted positron tracer for tumors, a preparation method and application thereof, wherein radioactive nuclides are carried out on the KRAS G12C mutation targeted positron tracer based on AMG510 18 F labeling, and constructing a positron tracer of the target tumor KRAS G12C so as to solve the problem that the existing KRAS mutation detection means cannot completely meet the accurate diagnosis and treatment target.
In order to achieve the purpose, the technical scheme adopted by the invention is as follows:
providing tumor KRAS G12C mutation targeting compoundsIs composed of 18 F/ 19 F-QZLO with a chemical structure as follows:
wherein n is 0, 1, 2, 3, 4, 5 or 6.
In another aspect, there is provided a positron tracer comprising a tumor KRAS G12C mutation targeting compound as described herein, said positron tracer comprising said compound as 18 F-QZLO with a chemical structure as follows:
wherein n is 0, 1, 2, 3, 4, 5 or 6.
Further, the positron tracer also comprises a solvent.
Further, the solvent is PBS containing ethanol or water for injection or a physiological saline solution.
In another aspect, there is provided the use of a positron tracer as described for nuclide diagnosis of KRAS G12C mutation positive tumors.
In another aspect, there is provided a method of preparing the positron tracer as described, the method comprising:
dissolving a compound 3, namely AMG510, in dimethyl sulfoxide, adding a compound 1, namely an acetonitrile solution of ethylene glycol di-p-toluenesulfonate, adding anhydrous potassium carbonate and potassium iodide for reaction, separating a product by high performance liquid chromatography, and freeze-drying overnight to obtain a labeled precursor compound 5, namely QZLO-OTs;
preparation by means of an automated synthesis module containing 18 Positron tracers of F-QZLO.
Further, the preparation of the catalyst containing the catalyst by an automated synthesis module 18 The positron tracer of the F-QZLO specifically comprises the following steps:
dissolving anhydrous potassium carbonate in water to obtain a solution A, dissolving aminopolyether in acetonitrile to obtain a solution B, and mixing the solution A and the solution B to obtain a No. 1 reagent; dissolving a compound 5, namely QZLO-OTs, in anhydrous acetonitrile to obtain a No. 2 reagent;
prepared from a cyclotron 18 F]The fluoride ion is passed through and adsorbed on the QMA anion exchange column, and then the fluoride ion is adsorbed on the QMA anion exchange column 18 F]F - Eluting with No. 1 reagent into a reaction bottle of a synthesis module, heating and maintaining under the protection of high-purity nitrogen or helium to remove water, and cooling;
adding the reagent No. 2 into the reaction bottle, heating and keeping to obtain a mixture C, diluting the mixture C with injection water after the temperature of the mixture C is reduced to room temperature to obtain a mixture D, passing the mixture D through a C18 reverse phase solid phase extraction column, and sequentially eluting adsorbates on the C18 reverse phase solid phase extraction column into an injector by using absolute ethyl alcohol and the injection water to obtain a mixed solution E;
separating the mixed solution E by a preparative liquid chromatography of a synthesizer by taking acetonitrile as a mobile phase;
passing the separated product through C18 reversed phase solid phase extraction column, washing with injection water, sequentially eluting the adsorbate on C18 reversed phase solid phase extraction column with anhydrous ethanol and injection water, filtering with sterilizing filter to obtain the final product 18 Positron tracers of F-QZLO.
In another aspect, there is provided the use of a tumor KRAS G12C mutation targeting compound as described herein 19 F-QZLO with a chemical structure as follows:
wherein n is 0, 1, 2, 3, 4, 5 or 6;
the above-mentioned 19 F-QZLO was used as a stable standard.
In another aspect, there is provided a method for preparing the tumor KRAS G12C mutation targeting compound 19 F-QZLO, said 19 The preparation method of the F-QZLO comprises the following steps:
adding a compound 1, namely ethylene glycol di-p-toluenesulfonate and tetrabutylammonium fluoride into acetonitrile, adding anhydrous potassium carbonate and potassium iodide for reaction, separating a product by a high performance liquid chromatography, and freeze-drying overnight to obtain a compound 2;
dissolving compound 3, namely AMG510 in dimethyl sulfoxide, adding acetonitrile solution of compound 2, adding anhydrous potassium carbonate and potassium iodide for reaction, separating a product by high performance liquid chromatography, and freeze-drying overnight to obtain a stable standard compound 4, namely 19 F-QZLO。
Compared with the prior art, the invention has the following beneficial effects:
the invention labels AMG510 with radionuclide 18 And F, constructing a PET molecular imaging probe which is used as a tumor KRAS G12C mutation targeted positron tracer, wherein experiments show that the PET molecular imaging probe has appropriate physicochemical and radiological properties and more ideal biological characteristics, can be used for PET imaging of KRAS G12C mutation tumors, and is beneficial to specific diagnosis of the tumors, selection of targeted therapeutic drugs and evaluation of curative effects.
The invention selects radionuclides 18 F marks AMG 510. Since the molecular skeleton has carbon element, radionuclide is generally used 11 Labeling with C can minimally alter the molecular structure to ensure its affinity and selectivity for the receptor, but does not require any modification 11 Short C half life (T) 1/2 20.3min), an online accelerator is needed, which is not favorable for clinical popularization; while 68 Metal nuclide labels such as Ga need to modify chelating groups such as DOTA, and the like, so that the molecular structure is greatly changed, and the affinity and the selectivity of the metal nuclide labels to receptors are influenced; thus, the difference between these species, 18 f has a suitable half-life (T) 1/2 109.8min) for sufficient time in the labeling preparation and clinical use processes, and after the hydroxyl end of the AMG510 is connected with the glycol or the polyethylene glycol chain, the end is labeled with the radionuclide 18 F has little influence on the molecular structure, can ensure the affinity and the selectivity of the F to a receptor, and meanwhile, the polyethylene glycol chain can reduce the fat solubility of the compound, thereby being beneficial to clinical application.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings of the embodiments can be obtained according to the drawings without creative efforts.
FIG. 1 is a HRMS plot of Compound 2 prepared in example 1 of the present invention.
FIG. 2 is a HRMS plot of Compound 4 prepared in example 1 of the present invention.
FIG. 3 is a HRMS plot of Compound 5 prepared in example 1 of the present invention.
FIG. 4 is a graph of a compound prepared in example 1 of the present invention 18 Schematic diagram of F-QZLO Allinone module label.
FIG. 5 is a photograph of a film prepared in example 1 of the present invention 18 F-QZLO Radioactive high performance liquid chromatography (Radio-HPLC) chart.
FIG. 6 is a photograph prepared in example 1 of the present invention 18 F-QZLO in vitro stability results.
FIG. 7 is a photograph of a film prepared in example 1 of the present invention 18 F-QZLO distribution in normal mice results are shown.
FIG. 8 is a photograph prepared in accordance with example 1 of the present invention 18 F-QZLO KRAS G12C mutation tumor-bearing mouse PET imaging result chart.
Detailed Description
To facilitate an understanding of the invention, the invention will now be described more fully with reference to the accompanying drawings. Preferred embodiments of the present invention are shown in the drawings. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete.
It should be noted that like reference numerals and letters refer to like items and, thus, once an item is defined in one embodiment, it need not be further defined and explained in subsequent embodiments. The specific implementation processes are described in steps in certain embodiments, are only for clear and accurate expression, and are not to be construed as limiting the sequence. In addition, in the process described in the examples, the chemicals used in the steps are all available substances or commercial products.
AMG510 (sotoraib) is a small molecule, specific, irreversible inhibitor of KRAS G12C that specifically binds irreversibly to KRAS G12C protein to inhibit its activity, interferes with the dissociation of GDP from KRAS G12C protein, and inhibits KRAS-mediated signaling by locking KRAS G12C protein in an inactive GDP-bound state. Positron Emission Computed Tomography (PET) is a clinical examination imaging technology in the field of nuclear medicine, and relates to a marker in an examination process, and after the marker is injected into a human body, the condition of life metabolic activity is reflected through the aggregation of the marker in metabolism, so that the purpose of diagnosis is achieved. At present, no report on radionuclide labeling of AMG510 is found, and the AMG510 is not used as a positron tracer for PET imaging to carry out radioactive diagnosis on KRAS G12C mutation positive tumors.
The invention connects ethylene glycol or polyethylene glycol chain on the hydroxyl end of AMG510 and marks radionuclide 18 F, obtaining a tumor KRAS G12C mutation targeting compound, constructing a PET molecular imaging probe, and preparing a PET imaging positron tracer targeting the tumor KRAS G12C, so that the PET imaging positron tracer is feasible for PET examination based on AMG510 preparation, and the method is named as 18 F-Quinazolineone, Quinazolineone 18 F-QZLO having the chemical structure:
comprises 18 The positron tracer of the F-QZLO also comprises a solvent, and the solvent can adopt PBS containing ethanol or injection water or physiological saline solution and is added in corresponding steps in the preparation process. The positron tracer structure contains ligand AMG510 targeting KRAS G12C mutant protein and PEG chain for radionuclide labeling, so that the positron tracer structure can be used for nuclide diagnosis of KRAS G12C mutation positive tumors.
Comprises 18 Preparation of positron tracers for F-QZLOThe preparation method comprises the following steps:
dissolving a compound 3, namely AMG510, in dimethyl sulfoxide, adding a compound 1, namely an acetonitrile solution of ethylene glycol di-p-toluenesulfonate, adding anhydrous potassium carbonate and potassium iodide for reaction, separating a product by high performance liquid chromatography, and freeze-drying overnight to obtain a labeled precursor compound 5, namely QZLO-OTs;
preparation by means of an automated synthesis module containing 18 A positron tracer of F-QZLO, comprising:
dissolving anhydrous potassium carbonate in water to obtain a solution A, dissolving aminopolyether in acetonitrile to obtain a solution B, and mixing the solution A and the solution B to obtain a No. 1 reagent; dissolving a compound 5, namely QZLO-OTs, in anhydrous acetonitrile to obtain a No. 2 reagent;
prepared from a cyclotron 18 F]The fluoride ion is passed through and adsorbed on the QMA anion exchange column, and then the fluoride ion is adsorbed on the QMA anion exchange column 18 F]F - Eluting with No. 1 reagent into a reaction bottle of a synthesis module, heating and maintaining under the protection of high-purity nitrogen or helium to remove water, and cooling;
adding the reagent No. 2 into the reaction bottle, heating and keeping to obtain a mixture C, diluting the mixture C with injection water after the temperature of the mixture C is reduced to room temperature to obtain a mixture D, passing the mixture D through a C18 reverse phase solid phase extraction column, and sequentially eluting adsorbates on the C18 reverse phase solid phase extraction column into an injector by using absolute ethyl alcohol and the injection water to obtain a mixed solution E;
separating the mixed solution E by a preparative liquid chromatography of a synthesizer by taking acetonitrile as a mobile phase;
passing the separated product through C18 reversed phase solid phase extraction column, washing with injection water, sequentially eluting the adsorbate on C18 reversed phase solid phase extraction column with anhydrous ethanol and injection water, filtering with sterilizing filter to obtain the final product 18 Positron tracers of F-QZLO.
The invention also provides another KRAS G12C mutation targeting compound 19 F-Quinazolineone ( 19 F-QZLO) by 18 And F mark as stable standard with the chemical structure:
19 the preparation method of the F-QZLO comprises the following steps:
adding a compound 1, namely ethylene glycol di-p-toluenesulfonate and tetrabutylammonium fluoride into acetonitrile, adding anhydrous potassium carbonate and potassium iodide for reaction, separating a product by a high performance liquid chromatography, and freeze-drying overnight to obtain a compound 2;
dissolving compound 3, namely AMG510, in dimethyl sulfoxide, adding acetonitrile solution of compound 2, adding anhydrous potassium carbonate and potassium iodide for reaction, separating the product by high performance liquid chromatography, and freeze-drying overnight to obtain stable standard compound 4, namely 19 F-QZLO。
The general formula of the two tumor KRAS G12C mutation targeting compounds is as follows:
the structure comprises a KRAS G12C targeting ligand AMG510, a glycol or polyethylene glycol (PEG) chain, and a labeled nuclide 18 F/ 19 F. Wherein n is 0, 1, 2, 3, 4, 5 or 6.
Both compounds can be prepared by the same synthetic route, 18 F/ 19 the synthetic route of the F-QZLO is as follows:
step 1: the compound 1 is ethylene glycol di-p-toluenesulfonate, the compound 1 is subjected to reaction conditions 'a' to obtain a compound 2, and the reaction conditions 'a' are as follows: TBAF, K 2 CO 3 ，ACN，KI，90℃。
n is 0, 1, 2, 3, 4, 5 or 6.
The step 1 specifically comprises the following steps:
Step 2: compound 3 is AMG510, compound 3 undergoes reaction condition "b" to give compound 4, and reaction condition "b" is: compound 2, K 2 CO 3 ，KI，DMSO，ACN，90℃。
n is 0, 1, 2, 3, 4, 5 or 6.
The step 2 specifically comprises the following steps:
And step 3: compound 3AMG510, compound 3 was subjected to reaction condition "c" to give compound 5, reaction condition "c" was: compound 1, K 2 CO 3 ，KI，ACN，DMSO，90℃。
n is 0, 1, 2, 3, 4, 5 or 6.
The step 3 specifically comprises the following steps:
dissolving the compound 3 in dimethyl sulfoxide, adding 2 times of the acetonitrile solution of the compound 1, adding 3 times of the anhydrous potassium carbonate and a trace amount of KI, and stirring at 90 ℃ for 12 hours. The product was isolated by HPLC, and lyophilized overnight to give labeled precursor compound 5, designated QZLO-OTs.
And 4, step 4: compound 5 is obtained by reaction conditions "d 18 F-QZLO, reaction conditions "d" are: [ 18 F]F - ， Kryptofix 2.2.2，K 2 CO 3 ，ACN，10min，100℃。
n is 0, 1, 2, 3, 4, 5 or 6.
The step 4 specifically comprises the following steps:
step 4.1: dissolving anhydrous potassium carbonate in water to obtain a solution A, dissolving aminopolyether (Kryptofix 2.2.2) in acetonitrile to obtain a solution B, and mixing the solution A and the solution B to obtain a No. 1 reagent; dissolving the compound 5 in anhydrous acetonitrile to obtain a No. 2 reagent;
step 4.2: the term "prepared by using a cyclotron 18 F]The fluoride ion is passed through and adsorbed on the QMA anion exchange column, and then the fluoride ion is adsorbed on the QMA anion exchange column 18 F]F - Eluting with reagent No. 1 into a reaction bottle of a synthesizer, heating to 60 deg.C under the protection of high-purity nitrogen or helium gas, maintaining for 1.0min, heating to 85 deg.C, maintaining for 2.0min, heating to 120 deg.C, maintaining for 2.0min, and cooling to 60 deg.C;
step 4.3: adding the reagent No. 2 into the reaction bottle, heating to 100 ℃, keeping the temperature for 10.0min to obtain a mixture C, diluting the mixture C with injection water after the mixture C is cooled to room temperature to obtain a mixture D, passing the mixture D through a C18 reverse phase solid phase extraction column, and then sequentially eluting adsorbates on the C18 reverse phase solid phase extraction column into an injector by using absolute ethyl alcohol and the injection water to obtain a mixed solution E;
step 4.4: separating the mixed solution E by a preparative liquid chromatography of a synthesizer by taking 45% acetonitrile as a mobile phase;
step 4.5: diluting the separated product with injection water, passing through C18 reversed phase solid phase extraction column, washing with injection water, eluting with anhydrous ethanol and injection water sequentially, filtering with sterilizing filter to obtain extract containing C18 reversed phase solid phase 18 Positron tracers of F-QZLO.
Example 1:
preparation of a composition comprising
I.e. having a tetrapolyethylene glycol chain 18 Positron tracer of F-QZLO:
step 1: compound 1, tetraethylene glycol di-p-toluenesulfonate (1.05g, 2.09mmol), tetrabutylammonium fluoride (TBAF, 0.55g, 2.10mmol), anhydrous potassium carbonate (K) 2 CO 3 0.89g, 6.44mmol) and a trace of KI) were added to 2mL of acetonitrile and stirred under reflux at 90 ℃ for 12 hours. The product was isolated by HPLC and lyophilized overnight to give compound 2 as a yellow oil (0.51mg, 69.6%). HRMS (ESI-Tof), [ M + H] + :m/z calculated for C 15 H 24 FO 6 S + 351.1272,found 351.0597。
Step 2: compound 3, AMG510(3, 26.2mg, 0.047mmol) was dissolved in 0.2mL of anhydrous DMSO and 0.8mL of compound 2(27.6mg, 0.079mmol) in acetonitrile was added to form a light yellow clear solution. After anhydrous potassium carbonate (25.9mg, 0.19mmol) was added, the yellow color of the solution was deepened, and the reaction was refluxed for 12 hours. Purification by HPLC to give the product, freeze-drying overnight to give compound 4 as a white powder, a stable standard: ( 19 F-QZLO,13.8mg，39.8％)。HRMS(ESI-Tof),[M+ H] + :m/z calculated for C 38 H 46 F 3 N 6 O 6 + 739.3425,found 739.1376。
And step 3: compound 1(136.0mg, 0.28mmol) and compound 3(81.2mg, 0.14mmol) were synthesized in the same manner as Compound 4 to give the labeled precursor compound 5 as a white solid, i.e., a labeled precursor (QZLO-OTs,43.0mg, 15.5%). HRMS (ESI-Tof), [ M + H] + :m/z calculated for C 45 H 52 F 2 N 6 O 9 S + 890.3485,found 891.1248。
And 4, step 4: 18 F-QZLO was prepared by the Trasis Allinone automated synthesis module.
Step 4.1: 3.0mg of potassium carbonate (K) 2 CO 3 ) Dissolving in 0.2mL of water to obtain solution A, and adding 13.0mg of aminopolyether (Kryptofix 2.2.2, K) 2.2.2 ) Dissolving in 0.8mL Acetonitrile (ACN) to obtain a solutionB, mixing the solution A and the solution B to obtain a No. 1 reagent;
2.0mg of QZLO-OTs was dissolved in 1.0mL of anhydrous acetonitrile to obtain reagent No. 2. The QZLO-OTs are compounds having the structure shown as follows:
step 4.2: prepared from a cyclotron 18 F]Fluorine ion ([ alpha ]) 18 F]F - 300mCi, 2.5mL) was passed through and adsorbed on the QMA anion exchange column, and then the solution was adsorbed on the QMA anion exchange column 18 F]F - Elute with reagent No. 1 into the reaction flask of the synthesizer, then under high purity nitrogen (N) 2 ) Heating to 60 deg.C under protection and maintaining for 1.0min, heating to 85 deg.C and maintaining for 2.0min, heating to 120 deg.C and maintaining for 2.0min, and cooling to 60 deg.C;
step 4.3: adding the reagent No. 2 into the reaction bottle, heating to 100 ℃, keeping the temperature for 10.0min to obtain a mixture C, diluting the mixture C to 15 mL by using water for injection after the mixture C is cooled to room temperature to obtain a mixture D, passing the mixture D through a C18 reverse phase solid phase extraction column, and then sequentially eluting adsorbates on the C18 reverse phase solid phase extraction column into a syringe by using 2.0mL of absolute ethyl alcohol (anhydrous ethanol) and 2.0mL of water for injection to obtain a mixed solution E;
step 4.4: separating the mixed solution E by using 45% acetonitrile as a mobile phase at the flow rate of 4mL/min through a preparative liquid chromatogram of a synthesizer, and collecting a separation product with the retention time of 21-23 min on a gamma chromatogram in a 50 mL transfer bottle (containing 25mL of injection water);
step 4.5: passing the separated product through C18 reverse phase solid phase extraction column, washing with 10mL injection water, sequentially eluting the adsorbate on the reverse phase solid phase extraction column with 1.5mL anhydrous ethanol (ethanol) and 13.5mL injection water, filtering with sterilizing filter (Merck Millipore Milllex-GV 0.22 μm SLGVR33RB) to obtain injectable product 18 F-QZLO solution (10% ethanol).
The preparation time of this example was about 90 minutes, the total radiochemical yield was 21.53 ± 1.46% (n-3, corrected for decay) and the radiochemical purity was greater than 99%.
The performance of the positron tracer prepared in this example was determined as follows:
(1) radioactive high performance liquid chromatography (Radio-HPLC) identification:
HPLC conditions are as follows: the column was an octadecyl bonded silica gel reverse phase column (Inertsil ODS-SP,4.6 mm. times.250 mm,5 μm, Shimadzu Corp.) and the mobile phase was acetonitrile and water (v: v. 50: 50), and the flow rate was 1 mL/min. 18 The retention time of F-QZLO is 14.5min, the radiochemical purity is more than 99%, and is higher than that in pharmacopeia 18 The radiochemical purity of F-deoxyglucose is higher than the specified standard of 90% (2020 edition of Chinese pharmacopoeia, second part). The Radio-HPLC results are shown in FIG. 5.
(2) And (4) checking:
the pH value is 5.0-8.0 (China pharmacopoeia 2020 edition, second part, appendix VI H). And (3) detecting bacterial endotoxin: taking appropriate amount of the product (i.e. filtering with a sterilizing filter to obtain the product for injection 18 F]QZLO solution), diluted 60 times with water for bacterial endotoxin test, and tested according to standard methods (chinese pharmacopoeia 2020 edition, section two, appendix XI E), with endotoxin content per 1mL less than 15 EU. And (4) sterile inspection: a proper amount of the product is taken and detected according to a standard method (China pharmacopoeia 2020 edition, second part, appendix XI H), and the product meets the requirements.
(3) The radioactive concentration: accurately measuring a certain volume of the product, placing the product in an activity meter to measure activity, and calculating the radioactive concentration according to the volume and the activity of the sample. The radioactive concentration of the product is more than 110 MBq/mL.
(4) The validity period is as follows: 6h is calculated from the calibration time.
(5) Fat solubility: measured by the shake flask method 18 The liposolubility of F-QZLO is log D of 2.17 +/-0.08.
(6) Normal mice in vivo distribution: FIG. 7 is a schematic view of 18 F-QZLO distribution in normal mice results are shown. The results show that it is possible to display, 18 the uptake value of F-QZLO is similar in each organ, and the F-QZLO is mainly discharged out of the body through intestinal metabolism. And can see 18 F-QZLO showed no significant in vivo defluorination and was injected at 2 hours post bone uptake of about 4% ID/g.
(7) PET imaging of KRAS G12C mutant tumor-bearing mice: in H358 xenografted mice, dynamic PET/CT imaging was performed for 1 hour. As shown in fig. 7, tumors were observed immediately after the administration, and the uptake thereof was 3.98% ID/g and slightly increased with the lapse of time. This indicates that 18 F-QZLO reaches the tumor site with the blood immediately after tail vein injection and is taken up by irreversible binding to Cys 12. Meanwhile, the uptake of muscle also increased with the passage of time, the ratio of tumor to muscle (T/M) reached a maximum value at 5 minutes (T/M2.27), and the ratio of tumor to muscle (T/M) remained substantially at about 1.7 after 30 minutes, indicating that 18 The distribution of F-QZLO reaches equilibrium 30 minutes after intravenous injection. We can also see from the results of the imaging, 18 F-QZLO has higher initial intake in the liver, is rapidly metabolized and excreted through bile and intestinal tracts, and is consistent with the distribution result in normal mice.
FIG. 6 is a drawing showing 18 In vitro stability results of F-QZLO after incubation for 2h at RT 18 The radiochemical purity of F-QZLO in physiological saline is still more than 99 percent, which shows that 18 The F-QZLO has high physiological saline stability and is favorable for clinical use.
The present invention has been described in terms of specific examples, which are provided to aid understanding of the invention and are not intended to be limiting. For a person skilled in the art to which the invention pertains, several simple deductions, modifications or substitutions may be made according to the idea of the invention.
Claims (9)
1. Tumor KRAS G12C mutation targeting compound characterized by:
the compound is 18 F/ 19 F-QZLO having the chemical structure:
wherein n is 0, 1, 2, 3, 4, 5 or 6.
2. A positron tracer comprising a tumor KRAS G12C mutation targeting compound as claimed in claim 1 wherein:
the positron tracer contains the compound 18 F-QZLO having the chemical structure:
wherein n is 0, 1, 2, 3, 4, 5 or 6.
3. The positron tracer of claim 2, wherein:
the positron tracer further comprises a solvent.
4. A positron tracer according to claim 3, wherein:
the solvent is PBS containing ethanol or water for injection or physiological saline solution.
5. Use of a positron tracer according to claim 4 wherein:
the positron tracer is used for nuclide diagnosis of KRAS G12C mutation positive tumors.
6. The method for preparing a positron tracer according to claim 4, wherein:
the preparation method comprises the following steps:
dissolving a compound 3, namely AMG510, in dimethyl sulfoxide, adding a compound 1, namely an acetonitrile solution of ethylene glycol di-p-toluenesulfonate, adding anhydrous potassium carbonate and potassium iodide for reaction, separating a product by high performance liquid chromatography, and freeze-drying overnight to obtain a labeled precursor compound 5, namely QZLO-OTs;
preparation by means of an automated synthesis module containing 18 Positron tracers of F-QZLO.
7. The method for preparing a positron tracer according to claim 6, wherein:
preparation of a peptide containing 18 The positron tracer of the F-QZLO specifically comprises the following steps:
dissolving anhydrous potassium carbonate in water to obtain a solution A, dissolving aminopolyether in acetonitrile to obtain a solution B, and mixing the solution A and the solution B to obtain a No. 1 reagent; dissolving a compound 5, namely QZLO-OTs, in anhydrous acetonitrile to obtain a No. 2 reagent;
the term "prepared by using a cyclotron 18 F]The fluoride ion is passed through and adsorbed on the QMA anion exchange column, and then the fluoride ion is adsorbed on the QMA anion exchange column 18 F]F - Eluting with No. 1 reagent into a reaction bottle of a synthesis module, heating and maintaining under the protection of high-purity nitrogen or helium to remove water, and cooling;
adding the reagent No. 2 into the reaction bottle, heating and keeping to obtain a mixture C, diluting the mixture C with injection water after the temperature of the mixture C is reduced to room temperature to obtain a mixture D, passing the mixture D through a C18 reverse phase solid phase extraction column, and sequentially eluting adsorbates on the C18 reverse phase solid phase extraction column into an injector by using absolute ethyl alcohol and the injection water to obtain a mixed solution E;
separating the mixed solution E by a preparative liquid chromatography of a synthesizer by taking acetonitrile as a mobile phase;
passing the separated product through C18 reversed phase solid phase extraction column, washing with injection water, sequentially eluting the adsorbate on C18 reversed phase solid phase extraction column with anhydrous ethanol and injection water, filtering with sterilizing filter to obtain the final product 18 Positron tracers of F-QZLO.
8. The use of the tumor KRAS G12C mutation targeting compound of claim 1, wherein:
the compound is 19 F-QZLO having the chemical structure:
wherein n is 0, 1, 2, 3, 4, 5 or 6;
the described 19 F-QZLO was used as a stable standard.
9. The method for preparing a tumor-targeting KRAS G12C mutant compound according to claim 1, wherein the compound is selected from the group consisting of:
the compound is 19 F-QZLO, said 19 The preparation method of the F-QZLO comprises the following steps:
adding a compound 1, namely ethylene glycol di-p-toluenesulfonate and tetrabutylammonium fluoride into acetonitrile, adding anhydrous potassium carbonate and potassium iodide for reaction, separating a product by a high performance liquid chromatography, and freeze-drying overnight to obtain a compound 2;
dissolving compound 3, namely AMG510 in dimethyl sulfoxide, adding acetonitrile solution of compound 2, adding anhydrous potassium carbonate and potassium iodide for reaction, separating a product by high performance liquid chromatography, and freeze-drying overnight to obtain a stable standard compound 4, namely 19 F-QZLO。
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