CN116375709A - Folic acid receptor targeting drug, metal complex, preparation method and application thereof - Google Patents

Folic acid receptor targeting drug, metal complex, preparation method and application thereof Download PDF

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CN116375709A
CN116375709A CN202211669071.1A CN202211669071A CN116375709A CN 116375709 A CN116375709 A CN 116375709A CN 202211669071 A CN202211669071 A CN 202211669071A CN 116375709 A CN116375709 A CN 116375709A
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cancer
folic acid
acid
metal complex
folate receptor
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杨大参
邵国强
杨凌云
罗志刚
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Nanjing Pet Tracer Co ltd
Nanjing First Hospital
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Nanjing Pet Tracer Co ltd
Nanjing First Hospital
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D475/00—Heterocyclic compounds containing pteridine ring systems
    • C07D475/02—Heterocyclic compounds containing pteridine ring systems with an oxygen atom directly attached in position 4
    • C07D475/04—Heterocyclic compounds containing pteridine ring systems with an oxygen atom directly attached in position 4 with a nitrogen atom directly attached in position 2
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
    • C07B59/00—Introduction of isotopes of elements into organic compounds ; Labelled organic compounds per se
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
    • C07B2200/00—Indexing scheme relating to specific properties of organic compounds
    • C07B2200/05—Isotopically modified compounds, e.g. labelled

Abstract

The invention provides a folic acid receptor targeting drug, a metal complex, a preparation method and application thereof, and relates to the technical field of radiolabeling. The folic acid receptor targeting drug has a structure shown as a formula I, and a metal complex is prepared after the folic acid receptor targeting drug is marked by a radionuclide. The folic acid receptor targeted drug or the metal complex provided by the invention can be used for diagnosing and/or treating diseases with over-expressed folic acid receptors. Has the advantages of high in vivo stability, strong specificity, good targeting property and the like.

Description

Folic acid receptor targeting drug, metal complex, preparation method and application thereof
Cross Reference to Related Applications
The present application claims priority from chinese patent application No. cn202111647679.X, entitled "a folate receptor targeted drug, metal complex, and methods of making and using thereof," filed at 12 months 30 of 2021, the entire contents of which are incorporated herein by reference.
Technical Field
The invention relates to the technical field of radiolabeling, in particular to a folic acid receptor targeting drug, a metal complex, a preparation method and application thereof.
Background
Folate Receptor (FR) is a glycoprotein linked to the cell membrane by the glycan phosphoinositides. FR has a high affinity and specificity for folic acid and its derivatives, such as methyltetrahydrofolate. FR mainly includes the α, β, γ and δ4 subtypes. Fα is the predominant folate receptor and is expressed restrictively in normal tissues, but is overexpressed in tumor tissues of epithelial origin, and the expression levels increase with progression of the tumor. Drugs or other substances that bind to folic acid can act by entering cells through fra-mediated endocytosis. Monoclonal antibodies targeting the folate receptor have good prospects in diagnosis and treatment of tumors.
In recent years, there has been an increasing search for radionuclide-labeled folic acid chelate imaging agents. 111 In-DTPA-folate is the first FR tumor imaging agent to enter and complete a second phase clinical trial, and better distinguishes benign and malignant tumors, but is less desirable In diagnosing recurrent ovarian cancer and endometrial malignant solid tumors (Journal of Nuclear Medicine,2003 (44): 700-707). EC20 is a peptide containing structural analogues of folic acid, 99m Tc-EC20 has higher uptake in tumor cells, shorter half-life, and can be protogenic in the kidneyNo further progress was made after the completion of the primary and secondary clinical trials of ovarian, cervical and renal cancers, etc., in 2015 (Journal of Nuclear Medicine,2004 (45): 857-866). 67/68 Ga-DOTA-fostate has limited clinical application due to poor in vivo stability (Eur J Nucl Med Mol Imaging,2011, 38:108-119). 18 F-click-fonate may show specific uptake in tumor parts where FR is highly expressed, but affects the imaging effect due to excessive abdominal uptake (bioconjugate Chemistry,2008,19 (12): 2462-2470). Currently, most FR-mediated radiopharmaceuticals are limited in their research period, and drugs that enter the clinic are also terminated by certain limitations themselves.
In view of this, the present invention has been made.
Disclosure of Invention
Aiming at the defects of the prior art, one of the purposes of the invention is to provide a novel folic acid receptor targeting drug and a preparation method thereof.
The second object of the invention is to provide a metal complex and a method for preparing the same.
It is a further object of the invention to provide the use of a folate receptor targeted drug and/or a metal complex as described above.
The invention aims at realizing the following technical scheme:
the invention provides a folic acid receptor targeting drug which is a compound shown in the following formula I or an isomer thereof,
Figure BDA0004014433750000021
wherein L is selected from a substituted or unsubstituted aryl diamine, a substituted or unsubstituted heteroaryl diamine, a substituted or unsubstituted heterocyclic diamine, a substituted or unsubstituted amino alcohol, a substituted or unsubstituted alkyl glycol group, or a polyethylene glycol derivative of the foregoing; the L is connected with the two side structural fragments through a-NH, -N or-O group;
R 1 is a bifunctional chelating agent;
R 2 selected from hydrogen or methyl, or is empty;
R 3 hydrogen, or null.
In particular, the method comprises the steps of,
Figure BDA0004014433750000022
as folic acid molecular structural fragment, when R 2 When hydrogen or methyl, -NR 2 The ortho-carbon atom has chirality, and the configuration can be R, S or RS.
Further, the folic acid molecular structural fragment is selected from the following structures:
Figure BDA0004014433750000023
it is of course understood that the structures at both ends of L are-NH, -N, -O, respectively, i.e., each of the ends of L is independently selected from imino groups, nitrogen atoms, or oxygen atoms.
Further, the L is selected from the following structures:
Figure BDA0004014433750000031
any one of the groups m, n, o, p and q is a natural number of 1 to 9.
Preferably, said L is selected from
Figure BDA0004014433750000032
More preferably, said L is
Figure BDA0004014433750000033
The bifunctional chelating agent is selected from iminodiacetic acid, ethylenediamine tetraacetic acid (EDTA), triethylenetetramine (TETA), diethylenetriamine-N, N, N ', N ', N ' -pentaacetic acid (DTPA), bis- (carboxymethyl imidazole) glycine, 6-hydrazinopyridine-3-carboxylic acid (HYNIC), N, N ' -bis [ 2-hydroxy-5- (carboxyethyl) benzyl ] ethylenediamine-N, any one of N "-diacetic acid (HEBD-CC), N ' - {5- [ acetyl (hydroxy) amino ] pentyl } -N- [5- ({ 4- [ (5-aminopentyl) (hydroxy) amino ] -4-oxobutanoyl } amino) pentyl ] -N-hydroxysuccinamide (DFO), 1,4,7, 10-tetraazacyclododecane-N, N ', N ' -tetraacetic acid (DOTA), 2- (4, 7, 10-tris (carboxymethyl) -1,4,7, 10-tetraazacyclododecane-1-yl) glutaric acid (dotga), 1,4, 7-triazacyclononane-1, 4, 7-triacetic acid (NOTA), 2- (4, 7-bis (carboxymethyl) -1,4, 7-triazo-1-yl) glutaric acid (nodga) and derivatives thereof.
Preferably, the bifunctional chelating agent is selected from DOTA or NOTA.
Specifically, the folic acid receptor targeting drug is selected from any one of compounds shown in the following structural formulas:
Figure BDA0004014433750000034
Figure BDA0004014433750000041
Figure BDA0004014433750000051
the embodiment of the invention also provides a preparation method of the folic acid receptor targeting drug by a solid phase synthesis method, and the folic acid receptor targeting drug can be specifically targeted and combined with FR alpha.
The embodiment of the invention also provides a metal complex which comprises the radionuclide and the folic acid receptor targeting drug. The radionuclide is selected from Al 18 F]、 64 Cu、 67 Cu、 67 Ga、 68 Ga、 89 Zr、 86 Y、 90 Y、 99m Tc、 111 In、 153 Sm、 166 Ho、 177 Lu、 186 Re、 188 Re、 211 At、 212 Bi、 213 Bi、 225 Ac、 227 Any one of Th. The biological distribution and targeting of the probe in vivo are examined by a small animal testAnd specificity.
The embodiment of the invention also provides a pharmaceutically acceptable composition, which comprises the folic acid receptor targeted drug or the metal complex, or pharmaceutically acceptable salts thereof and pharmaceutically acceptable excipients.
The embodiment of the invention also provides application of the folic acid receptor targeted drug or the metal complex or the pharmaceutically acceptable composition in diagnosis and/or treatment of folic acid receptor over-expression diseases.
Further, the diseases are brain tumor, breast cancer, ovarian cancer, gastric cancer, lung cancer, kidney cancer, mesothelial tissue cancer, endometrial cancer, colorectal cancer, cervical cancer, head and neck tumor and testicular cancer.
The invention has the following beneficial effects: the folic acid receptor targeting drug provided by the embodiment of the invention consists of a folic acid molecular structure fragment, a linking agent L and a bifunctional chelating agent R 1 The composition is formed. In order not to change the affinity with FR, the invention can regulate the residence time of the compound in vivo by introducing a linking agent L on the basis of folic acid medicine structure, and high tumor specific uptake is realized after radionuclide labeling. The metal complex provided by the invention is mainly excreted through kidneys and can be used for diagnosing or treating tumors of targeted folate receptors. The folic acid receptor targeting drug provided by the invention can specifically target the FR alpha receptor, and the preparation method is simple and convenient; the metal complex has high labeling rate, good in vivo and in vitro stability, strong specificity and higher diagnosis and treatment efficacy.
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In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings that are needed in the embodiments will be briefly described below, it being understood that the following drawings only illustrate some embodiments of the present invention and therefore should not be considered as limiting the scope, and other related drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is an LC-MS spectrum of Compound A provided in example 1 of the present invention;
FIG. 2 is a schematic diagram of embodiment 7 of the present invention 68 A radioactive spectrum of Ga-1;
FIG. 3 is a diagram of embodiment 10 of the present invention 68 Ga-1 biodistribution in ICR mice;
FIG. 4 is a diagram of embodiment 11 of the present invention 68 microPET imaging of Ga-1 in SKOV3 tumor-bearing mice;
FIG. 5 is a view of Al [ in accordance with example 12 of the present invention 18 F]-1 microPET imaging of tumor-bearing mice at SKOV 3;
FIG. 6 is a schematic illustration of an embodiment 13 of the present invention 68 microPET imaging of Ga-2 in SKOV3 tumor-bearing mice;
FIG. 7 is a schematic illustration of an embodiment 14 of the present invention 68 microPET imaging of Ga-1 in SKOV3 tumor-bearing mouse blocking test;
FIG. 8 is a schematic diagram of an embodiment 15 of the present invention 68 microPET imaging of Ga-1 in PC-3 tumor-bearing mice.
Detailed Description
The features and capabilities of the present invention are described in further detail below in connection with the examples. The specific conditions are not noted in the examples and are carried out according to conventional conditions or conditions recommended by the manufacturer. The reagents or apparatus used were conventional products commercially available without the manufacturer's attention.
The features and capabilities of the present invention are described in further detail below in connection with the examples.
Example 1
The embodiment provides a folic acid receptor targeting drug A, the structural formula of which is shown as follows:
Figure BDA0004014433750000061
the embodiment also provides a preparation method of the folic acid receptor targeting drug A, which is prepared by referring to the following synthetic route:
Figure BDA0004014433750000071
the specific operation is as follows:
and (3) preparing resin: the wang resin (1.00 mmol,1.04 eq), fmoc-Glu (OAlly) -OH (3.00 mmol,3.00 eq), HOAT (3.00 mmol,3.00 eq), DIC (3.00 mmol,3.00 eq) and DMAP (0.30 mmol,0.3 eq) were dissolved in DMF (5.00 mL). The mixture was heated at 20deg.C, N 2 Stirring for 3h under the condition. Then Ac is added 2 O/DIEA/DMF (10/5/85) 5.00mL in N 2 Stirring was continued for 15min under the conditions. The resin was washed with DMF (20.0 ml x 5) and the mixture was filtered to give resin 1.
Deprotection: DMF solution (20% piperidine, 40.0 mL) was added to resin 1 above and the mixture was taken under N 2 Stirring for 20min under the condition. Then washed with DMF (40.0 ml x 5) and filtered.
Coupling: a solution of pteroic acid (1.50 eq) in DMSO (6 mg/ml), a solution of HOAT (3.00 eq) and EDCI (3.00 eq) in DIEA (3.00 eq) was added to the resin at 20℃N 2 Stirring is carried out for 12h under the condition. Washed with DMF (40.0 ml x 5) and filtered to give resin 2.
Deprotection: phSiH (10.0 eq) and Pd (PPh) 3 ) 4 DMF (40.0 mL) solution of (B) was added to the above resin 2 at N 2 Stirring for 1h, washing with DMF (40.0 ml 5) and then filtering gave resin 3.
Coupling: a solution of piperazine (3.00 eq) with single Boc protection in DMSO (6 mg/ml), HOAT (2.00 eq) and DIEA (0.5 ml) in DIC (2.00 eq) was added to resin 3 at 20℃N 2 Stirring is carried out for 12h under the condition. Washed with DMF (40.0 ml x 5) and filtered to give resin 4.
Cracking: resin 4 was washed 3 times with methanol and dried in vacuo. Then treated with 5mL of TFA for 45min. The peptide was precipitated with cold isopropyl ether, centrifuged (3000 rpm,2 min) and washed 2 times with isopropyl ether. The crude peptide was dried under vacuum for 2h. Preparation of crude peptide using high performance liquid chromatography (A: H 2 O (0.075% TFA), B: ACN) to give compound 4' (0.08 g,119 umol) as a yellow solid in yield: 11.9%, purity: 93%.
1 H NMR(500MHz,Chloroform-d)δ8.47(t,J＝6.0Hz,1H),7.71–7.65(m,2H),6.81–6.74(m,2H),6.59(d,J＝7.1Hz,1H),6.27(d,J＝7.1Hz,1H),4.94(dd,J＝8.9,2.1Hz,2H),4.33(q,J＝11.7Hz,1H),3.57(ddd,J＝6.4,3.8,1.1Hz,4H),2.98–2.89(m,4H),2.50–2.34(m,2H),1.87–1.76(m,2H)。
Coupling: a mixture of compound 4' (0.08 g,119.32umol,1 eq) and DIEA (74.2 mg, 514 umol,0.1mL,4.81 eq) was dissolved in DMSO (8.00 mL) and stirred at 25℃for 0.5h, followed by the addition of 2- [4- (carboxymethyl) -7- [2- (4-nitrophenoxy) -2-oxo-ethyl ] -1,4, 7-triazol-1-yl ] acetic acid (NOTA-PNP) (55.7 mg,131umol,1.1 eq). Stirring was continued for 1h at 25 ℃. The reaction was purified by preparative HPLC (TFA) to give product a (22.0 mg,26.9 umol) as a yellow solid in 22.5% yield with 97.5% HPLC purity.
LCMS：398.4[M/2+H] + ；795.5[M+H] + 。
1 H NMR(500MHz,Chloroform-d)δ8.47(s,1H),7.74–7.68(m,2H),6.82–6.75(m,2H),6.59(d,J＝7.1Hz,1H),6.27(d,J＝6.9Hz,1H),4.99(d,J＝8.9Hz,2H),4.32(q,J＝11.7Hz,1H),3.53(s,6H),3.58–3.48(m,2H),3.46(d,J＝0.9Hz,4H),3.17(d,J＝4.1Hz,2H),2.60–2.50(m,12H),2.48–2.37(m,2H),1.83–1.70(m,2H)。
Example 2
The embodiment provides a folic acid receptor targeting drug B, the structure of which is as follows:
Figure BDA0004014433750000081
synthetic procedure referring to example 1, 2- [4- (carboxymethyl) -7- [2- (4-nitrophenoxy) -2-oxo-ethyl ] -1,4, 7-triazol-1-yl ] acetic acid (NOTA-PNP) of example 1 was replaced with 2,2',2"- (10- (2- (4-nitrophenoxy) -2-acetoxy) -1,4,7, 10-tetraazacyclododecane-1, 4, 7-tri-yl) triacetic acid (DOTA-PNP).
1 H NMR(500MHz,Chloroform-d)δ8.47(s,1H),7.72–7.65(m,2H),6.82–6.76(m,2H),6.59(d,J＝7.0Hz,1H),6.27(d,J＝7.1Hz,1H),4.99(d,J＝8.9Hz,2H),4.32(q,J＝11.7Hz,1H),3.58–3.48(m,8H),3.46(d,J＝0.7Hz,6H),3.17(d,J＝4.0Hz,2H),2.56(d,J＝1.8Hz,
16H),2.48–2.37(m,2H),1.83–1.70(m,2H)。
Example 3
The embodiment provides a folic acid receptor targeting drug C, the structure of which is as follows:
Figure BDA0004014433750000091
synthetic procedure referring to example 1, piperazine in example 1 was exchanged for 3- (2-hydroxyethoxy) -1- (piperazin-1-yl) propan-1-one.
1 H NMR(500MHz,Chloroform-d)δ8.47(s,1H),7.72–7.65(m,2H),6.82–6.76(m,2H),6.62–6.51(m,1H),6.27(d,J＝6.9Hz,1H),4.99(d,J＝8.9Hz,2H),4.38–4.26(m,3H),3.87–3.73(m,2H),3.61(td,J＝7.1,2.0Hz,2H),3.58–3.54(m,5H),3.54–3.42(m,7H),3.17(d,J＝4.1Hz,2H),2.61–2.50(m,14H),2.39–2.28(m,2H),2.09–1.96(m,2H)。
Example 4
The embodiment provides a folic acid receptor targeting drug D, the structure of which is as follows:
Figure BDA0004014433750000092
synthetic procedure reference is made to example 1, wherein the pteroic acid in example 1 is replaced by 4- [ [ [ (6R) -2-amino-3, 4,5,6,7, 8-hexahydro-4-oxo-6-pteridinyl ] methyl ] amino ] -benzoic acid.
1 H NMR(500MHz,Chloroform-d)δ7.76–7.70(m,2H),7.53(s,2H),6.65–6.58(m,2H),4.32(q,J＝11.7Hz,1H),3.68–3.58(m,1H),3.58–3.49(m,8H),3.49–3.42(m,7H),3.40–
3.34(m,1H),3.17(d,J＝4.1Hz,2H),2.60–2.50(m,12H),2.48–2.37(m,2H),1.83–1.70(m,2H)。
Example 5
The embodiment provides a folic acid receptor targeting drug E, the structure of which is as follows:
Figure BDA0004014433750000093
synthetic procedure referring to example 1, piperazine in example 1 was exchanged for 2, 5-dimethylamine-thiazole.
1 H NMR(500MHz,Chloroform-d)δ8.47(s,1H),7.73–7.67(m,2H),7.51(s,1H),6.82–6.76(m,2H),6.59(d,J＝7.1Hz,1H),6.27(d,J＝6.9Hz,1H),4.99(d,J＝8.9Hz,2H),4.77(dd,J＝8.0,0.9Hz,2H),4.55(dd,J＝8.7,0.7Hz,2H),4.32(q,J＝11.7Hz,1H),3.46(d,J＝1.0Hz,4H),3.21(s,2H),2.60–2.50(m,12H),2.38–2.30(m,1H),2.30–2.20(m,1H),1.79–1.65(m,2H)。
Example 6
The embodiment provides a folic acid receptor targeting drug F, the structure of which is as follows:
Figure BDA0004014433750000101
synthetic procedure reference is made to example 1, substituting pteroic acid from example 1 for 4- [ [ (6R) 2-amino-3, 4,5,6,7, 8-hexahydro-5-methyl-4-oxo-6-pteridinyl ] methyl ] amino ] -benzoic acid.
1 H NMR(500MHz,Chloroform-d)δ7.76–7.70(m,2H),7.61(s,2H),6.66–6.59(m,2H),4.32(q,J＝11.7Hz,1H),3.90–3.81(m,1H),3.63–3.42(m,16H),3.17(d,J＝4.1Hz,2H),2.92(d,J＝1.6Hz,3H),2.60–2.50(m,12H),2.48–2.37(m,2H),1.83–1.70(m,2H)。
Example 7
This example provides a folic acid metal complex 68 Ga-1, the structure of which is shown below:
Figure BDA0004014433750000102
taking an aqueous solution (20. Mu.L, 1 mg/mL) of Compound A prepared in example 1 and pretreatment 68 GaCl 3 Sodium acetate buffer solution (1.1 ml) of the above-mentioned materials are fully mixed, the pH of the reaction solution is regulated and kept between 4 and 4.5, the reaction solution is marked at room temperature, the radiochemical purity is 95.5%, and the HPLC radioactive spectrum is shown in figure 2.
And (3) identification:
standard substance preparationThe preparation method comprises the following steps: an aqueous solution (5 ml,10 mg/ml) of Compound A obtained in example 1 was taken and mixed with GaCl 3 Sodium acetate buffer solution (5 ml,20mg/ml, pH 4-4.5), and then reacted at room temperature for 24 hours. Purification by preparative HPC (YMC-Pack ODS-A-HG,10 μm, 150X 20mm,5ml/min,20% ethanol and 80% water for injection, UV 254 nm). The resulting solution was lyophilized to obtain a white solid powder.
HPLC: standard UV peak rt= 3.382min, radioactivity peak rt=3.785 min, consistent with standard peak position.
Will label the product 68 Ga-1 is respectively placed in mouse serum and normal saline, and after incubation for 12 hours in a 37 ℃ water bath box, the radiochemical purity is 94% and 95% respectively, which shows that the Ga-1 has good in vitro stability.
Example 8
This example provides a folic acid metal complex Al 18 F]-1, the structure of which is shown below:
Figure BDA0004014433750000111
2mM AlCl was prepared using 0.1mM acetate buffer pH 4.0 as solvent 3 A solution; 0.1mM acetic buffer, pH 4.0, 0.1mL AlCl was taken 3 6 mu L of solution is added with 50 to 120MBq 18 F - (0.1 mL) was allowed to stand at room temperature for 5 minutes, and then 5. Mu.L (1 mg/mL) of the solution of Compound A prepared in example 1 was added thereto, followed by reaction at 110℃for 10 minutes to give the objective compound. The target compound is separated and purified by Sep-pak C18 Column Light, before using, the Sep-pak Column is activated by 10mL of absolute ethyl alcohol and 10mL of high-purity water, and the target compound Al is eluted by 0.5mL of ethyl alcohol and 3mL of normal saline in sequence 18 F-1, its amplification purity was 94.4%.
And (3) identification:
and (3) preparation of a standard substance: to 0.5ml of a fluorine acetate solution were added the compound A (500. Mu.g) prepared in example 1, 0.8ml of acetonitrile and 0.05ml of 0.4M AlCl 3 The aqueous solution was reacted at 110℃for 10min, and the resulting product was isolated and purified by preparative HPLC.
HPLC: standard UV peak rt=4.129 min, radioactivity peak rt= 4.305min, consistent with standard peak position.
Will mark the product Al 18 F]-1 was placed in mouse serum and physiological saline respectively, and after incubation in 37 ℃ water bath for 2h, the radiochemical purity was 92.8% and 93.9% respectively, indicating good in vitro stability.
Example 9
This example provides a folic acid metal complex 68 Ga-2, the structure of which is shown below:
Figure BDA0004014433750000121
preparation of complexes according to reference example 7 68 Ga-2, compound A from example 7 is replaced by compound F, which is placed in a purity of 95.6%.
And (3) identification:
and (3) preparation of a standard substance: reference example 7 was made to the standard Ga-2, and compound A in example 7 was exchanged for compound F.
HPLC: standard UV peak rt= 4.677min, radioactivity peak rt= 4.830min, consistent with standard peak position.
Will label the product 68 Ga-2 is respectively placed in mouse serum and normal saline, and after incubation for 12 hours in a 37 ℃ water bath box, the radiochemical purity is 94.5% and 95.8% respectively, which shows that the Ga-2 has good in vitro stability.
Example 10
ICR mice were taken 20 animals, and were subjected to a leaf-acid-free diet for one week before the experiment, and randomly divided into 5 groups of 4 animals each by a lottery method, and 0.1ml (3.7 MBq) was injected into the tail vein respectively 68 Blood is taken through cardiac puncture and euthanized at 5min, 15min, 30min, 60min and 120min after Ga-1, important viscera are weighed by dissection, wet weight is weighed by dissection, radioactivity is counted through a gamma counter, time is recorded, radioactivity attenuation correction is carried out, radioactivity intake of each viscera, namely the percentage of radioactivity intake of unit mass tissue to injection dose (% ID/g) is calculated, and biological distribution is shown in figure 3.
Results： 68 Ga-1 is cleared more quickly in blood, and other than kidney, the radioactive uptake of heart, liver, spleen, lung and other normal tissues is lower, and the radioactive uptake (% ID/g) of most organs is reduced to below 2 at 120min, so that the medicine is mainly excreted through the kidney.
Example 11
Nude mice with human ovarian cancer (SKOV 3: FR alpha high expression) are 1, the age of the mice is 10-12 weeks, and the tumor length is 0.8cm. The tumor-bearing mice begin to take a free-leaf acid diet one week before imaging, and isoflurane is inhaled for anesthesia and tail vein injection 68 Ga-1 (3.7 MBq/0.1 mL) microPET imaging, image acquisition and processing parameters: the energy peak 350KeV-650KeV,timing window is 3.438ns, the acquisition time is 600s, the reconstruction method is OSEM3D/MAP iterative reconstruction, the tomographic image analysis and the delineation of tumor boundaries are carried out by using Inveon Research Workplace to obtain a region of interest (Regions of Interest, ROI), the radioactive uptake of the tumor (% ID/g) and the tumor/liver ratio are calculated, and the PET imaging result is shown in figure 4.
Results: 68 ga-1 is excreted mainly through urinary system (kidney and bladder), from 0.5-2.5 h, the radioactive uptake of tumor is continuously increased, the affinity to FR alpha is high, the imaging effect is good, and the imaging agent is excellent.
Example 12
microPET imaging of tumor-bearing mice with reference to example 11, tail intravenous injection in example 11 68 Ga-1 (3.7 MBq/0.1 mL) is changed to Al [ 18 F]-1 (3.7 MBq/0.1 mL), PET imaging results are shown in FIG. 5.
Results: intravenous injection of Al 18 F]After the medicine-1, the radioactive uptake of the tumor is continuously increased within 0.5-3.5 h, and the uptake of other non-target organs is lower. Description of Al [ 18 F]The-1 medicine is mainly excreted through the urinary system, has strong specificity and good imaging effect, and is a good imaging agent.
Example 13
microPET imaging of tumor-bearing mice with reference to example 11, tail intravenous injection in example 11 68 Ga-1 (3.7 MBq/0.1 mL) is exchanged for 68 Ga-2 (3.7 MBq/0.1 mL), PET imaging results are shown in FIG. 6.
Results: intravenous injection 68 Ga-2 medicineAfter the tumor uptake peaks in 0.5h, no obvious uptake occurs in 2.5h, and other non-target organs are cleared faster. Description of the invention 68 Ga-2 medicine is mainly excreted through urinary system, and has strong specificity and good imaging effect.
Example 14
microPET imaging of tumor-bearing mice with reference to example 11, tail intravenous injection in example 11 68 The PET imaging results are shown in FIG. 7, which shows that the normal folic acid-blocked FR (150. Mu.g/0.15 mL) dissolved in physiological saline was injected 20min before Ga-1 (3.7 MBq/0.1 mL).
Results: intravenous injection 68 After Ga-1 drug, the radioactive uptake of the tumor is extremely low in 0.5h, no obvious uptake is caused by the tumor in 1.5-2.5 h, and almost no uptake is caused by the tumor compared with the non-blocking group (example 11). Description of the invention 68 Ga-1 medicine can specifically target FR alpha in vivo and is a good imaging agent.
Example 15
microPET imaging was performed on tumor-bearing mice in accordance with example 11, and the results of PET imaging were shown in FIG. 8, with the nude mice of human ovarian cancer (SKOV 3: FR.alpha.high expression) in example 11 replaced with nude mice of human prostate cancer (PC 3: FR.alpha.low expression).
Results: intravenous injection 68 After Ga-1 medicine, no obvious uptake of tumor radioactivity exists in 0.5-2.5 h, and the uptake of other non-target organs except kidney and bladder is lower. Description of the invention 68 Ga-1 medicine is excreted mainly through urinary system, has strong specificity to FR alpha and is a good imaging agent.
The foregoing describes in detail preferred embodiments of the present invention. It should be understood that active modifications and variations can be made in accordance with the concepts of the invention by one of ordinary skill in the art without the need for creative efforts. Therefore, the technical solutions obtained by those skilled in the art through logic analysis, reasoning or limited experiments based on the prior art according to the present invention should be within the protection scope defined by the claims.

Claims (9)

1. A folic acid receptor targeting drug is characterized in that the folic acid receptor targeting drug is a compound shown in the following formula I or an isomer thereof,
Figure FDA0004014433740000011
wherein L is selected from a substituted or unsubstituted aryl diamine, a substituted or unsubstituted heteroaryl diamine, a substituted or unsubstituted heterocyclic diamine, a substituted or unsubstituted amino alcohol, a substituted or unsubstituted alkyl glycol group, or a polyethylene glycol derivative of the foregoing; the L is connected with the two side structural fragments through a-NH, -N or-O group;
R 1 is a bifunctional chelating agent;
R 2 selected from hydrogen or methyl, or is empty;
R 3 hydrogen, or null.
2. The folate receptor targeted drug of claim 1, wherein L is selected from the group consisting of
Figure FDA0004014433740000012
Any one of the groups m, n, o, p and q are natural numbers of 1-9;
the bifunctional chelating agent is selected from iminodiacetic acid, ethylenediamine tetraacetic acid (EDTA), triethylenetetramine (TETA), diethylenetriamine-N, N, N ', N ', N ' -pentaacetic acid (DTPA), bis- (carboxymethyl imidazole) glycine or 6-hydrazinopyridine-3-carboxylic acid (HYNIC), N, N ' -bis [ 2-hydroxy-5- (carboxyethyl) benzyl ] ethylenediamine-N, any one of N "-diacetic acid (HEBD-CC), N ' - {5- [ acetyl (hydroxy) amino ] pentyl } -N- [5- ({ 4- [ (5-aminopentyl) (hydroxy) amino ] -4-oxobutanoyl } amino) pentyl ] -N-hydroxysuccinamide (DFO), 1,4,7, 10-tetraazacyclododecane-N, N ', N ' -tetraacetic acid (DOTA), 2- (4, 7, 10-tris (carboxymethyl) -1,4,7, 10-tetraazacyclododecane-1-yl) glutaric acid (dotga), 1,4, 7-triazacyclononane-1, 4, 7-triacetic acid (NOTA), 2- (4, 7-bis (carboxymethyl) -1,4, 7-triazo-1-yl) glutaric acid (nodga) and derivatives thereof.
3. The folate receptor targeted drug of claim 2, selected from the following structural formulae:
Figure FDA0004014433740000021
Figure FDA0004014433740000031
4. the folate receptor targeted drug of any one of claims 1-3, wherein the folate receptor targeted drug is prepared using a solid phase synthesis method.
5. A metal complex comprising a radionuclide and the folate receptor targeted drug of any one of claims 1-3.
6. The metal complex of claim 5, wherein the radionuclide is selected from the group consisting of Al [ ] 18 F]、 64 Cu、 67 Cu、 67 Ga、 68 Ga、 89 Zr、 86 Y、 90 Y、 99m Tc、 111 In、 153 Sm、 166 Ho、 177 Lu、 186 Re、 188 Re、 211 At、 212 Bi、 213 Bi、 225 Ac、 227 Any one of Th.
7. A pharmaceutically acceptable composition comprising the folate receptor targeted drug of any one of claims 1-3, or the metal complex of claim 5 or 6, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.
8. Use of a folate receptor targeted drug according to any one of claims 1-3 or a metal complex according to claim 5 or 6 or a pharmaceutically acceptable composition according to claim 7 for the diagnosis and/or treatment of a folate receptor over-expression disease.
9. The use according to claim 8, wherein the disease is brain tumor, breast cancer, ovarian cancer, gastric cancer, lung cancer, renal cancer, mesothelial cancer, endometrial cancer, colorectal cancer, cervical cancer, head and neck tumor, testicular cancer.
CN202211669071.1A 2021-12-30 2022-12-23 Folic acid receptor targeting drug, metal complex, preparation method and application thereof Pending CN116375709A (en)

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