CN103880887B

CN103880887B - A kind of99mtcO-TYRDTC complex and its preparation method and application

Info

Publication number: CN103880887B
Application number: CN201410129878.5A
Authority: CN
Inventors: 张俊波; 朱菁菁; 胡其然; 杜进; 陆洁; 王学斌; 唐志刚
Original assignee: Beijing Normal University
Current assignee: Beijing Normal University
Priority date: 2014-04-02
Filing date: 2014-04-02
Publication date: 2016-08-24
Anticipated expiration: 2034-04-02
Also published as: CN103880887A

Abstract

The invention discloses one^99mTcO TYRDTC complex, it represents formula and is^99mTcO TYRDTC, by synthesis and the b. of a. part TYRDTC^99mTwo processing steps of the preparation of TcO TYRDTC are prepared from.^99mThe radiochemical purity height of TcO TYRDTC complex, good stability and parent's tumour function admirable, preparation simplicity, it is widely used in the tumor imaging of human body and animal, is a kind of novel amino acids tumor developer having application value.

Description

A kind of99mTcO-TYRDTC complex and its preparation method and application

Art

The present invention relates to^99mThe radiopharmaceutical chemistry of Tc mark and clinical nuclear medicine technical field, particularly relate to A kind of^99mTcO-TYRDTC complex and its preparation method and application.

Background technology

Malignant tumour is current one of principal disease endangering human health, and tumour annual morbidity is still presented with the death rate The trend of liter, early diagnosis of tumor and personalized treatment are significant for life cycle for the life and prolongation patient saving patient.Pass Lesion detection developing method such as x-ray method, CT, MRI etc. of non-wound of system, the mainly form of internal organs and structure under pathological state Make imaging, and radionuclide molecular image technology such as positron emission tomography (PET) and single photon emission tomography are swept Retouch art (SPECT) and the change of the physiology of tumour, pathology, metabolism and function can be reflected.Currently, along with PET/CT, SPECT/CT Play an increasingly important role in terms of early diagnosis of tumor and personalized treatment Deng organically blending of image technology, for adapting to Its extensively application in clinical diagnosis and treatment and fast-developing demand, rely interdependent " targeting ammunition " as this technology, newly Type the best highly sensitive Geigers probe, its research just seems particularly urgent and important.

For tumor developer,¹⁸F-fluorodeoxyglucose (¹⁸F-FDG) it is the widest tumour of current clinical practice Developer, enjoys " battle steed " and the good reputation of " century molecule ".¹⁸F-FDG PET imaging be now widely used for tumour diagnosis, By stages and therapeutic evaluation, but¹⁸F-FDG still has weak point, and its detection malignant tumour is the lowest.Due to¹⁸F-FDG absorbs also The non-tumor cell specific that is only absorbs, and some inflammatory cell, macrophage etc. also can absorb¹⁸, easily there is false positive in F-FDG Result causes false judgment.Therefore, a more preferable tumor developer of species specificity is still needed to clinically.

Due to some inflammatory cell, macrophage etc. will be less than right to amino acid whose picked-up^MakeThe picked-up of F-FDG, therefore puts Penetrating property isotope labeling amino acid being specifically better than as tumor developer¹⁸F-FDG.Research shows specific amino acid transporter egg In vain in many tumour cells, expression raises, and continues and cell out of control growth to meet in tumour, this just for diagnosis and Treatment provides potential target spot.The amino acid of radioisotope labeling can be as substrate targeting high expressed in tumour cell All kinds of amino acid transporters, therefore to amino acid or its modify afterproduct carry out radioactive label become specific tumour show An important directions as agent research.Up to now, existing 20 several amino acids movement systems are confirmed, and are wherein independent of In Na⁺Movement system L (LAT) of ion is the most deep.LAT movement system typically transports that size is relatively big, natural with side chain Amino acid (such as methionine etc.) or aromatics amino acid, such as tyrosine, phenylalanine and tryptophan etc..Up to now, radioactivity Mark amino acid and derivative, as S-(¹¹C-methyl)-L-Methionine (¹¹C-MET), O-(2-¹⁸F-fluoro ethyl)-L-junket ammonia Acid (¹⁸F-FET), have been used to clinical tumor imaging research, but¹¹C and¹⁸F needs to be produced by accelerator, expensive, and one Determine to hinder its extensive application in clinical diagnosis in degree, especially in countries and regions underdeveloped.With¹¹C With¹⁸F compares,^99mTc has a biggest superiority as medical science tracer nuclide: suitably half-life (T_1/2=6.02h), energy Moderate single photon γ (E γ=140keV) launches, decay daughter dose of radiation is low, and⁹⁹Mo-^99mBeing developed into of Tc generator Merit makes^99mThe preparation of Tc radiopharmaceutical is simple, can medicine box, cheap and easy to get, reliable in quality.Therefore, research^99mTc marks ammonia Base acid and derivative thereof are significant for tumor imaging, become a big focus of Today radiation drug development.

Due to^99mTcO mark gluceptate (^99mTcO-GH) be with [^99mTcO]³⁺Centered by the complex of core, structure Determine, and easily and the generation of other part generation ligand exchange reactions new [^99mTcO]³⁺Core marker ligand compound, so [^99mTcO]³⁺Caryogamy compound causes the extensive concern of people.At present,^99mThe preparation of TcO-GH has been realized in medicine box, for [^99mTcO]³⁺Radiopharmaceutic research and the application of core mark lay a solid foundation.Tyrosine is converted into its dithiocarbamic acid Salt part (Tyrosine dithiocarbamate, be called for short TYRDTC), utilize S atom in dithiocarbamate with^99mTcO caryogamy Position preparation^99mTcO-TYRDTC complex (^99mThe tyrosine dithiocarbamate of TcO core mark) come as novel^99mTc labeled amino acid Class tumor developer, is the vital task that currently faces of this area.

Summary of the invention

It is an object of the invention to provide one and prepare simplicity, radiochemical purity is high and close tumour function admirable^99mTcO-TYRDTC complex, also provides for its preparation method simultaneously.

In order to achieve the above object, the present invention is by the following technical solutions: a kind of^99mTcO-TYRDTC complex complex Preparation method, it represents formula and is^99mTcO-TYRDTC, described part TYRDTC structural formula is as follows:

^99mThe preparation method of TcO-TYRDTC is as follows:

A. the synthesis of part TYRDTC:

In reaction bulb, it is separately added into a certain amount of L-tyrosine, NaOH, stirs as solvent, ice-water bath using absolute methanol Mix dissolving, instill the CS being dissolved in ether₂, react 3 hours in ice-water bath after being added dropwise to complete, after filtering with sand core funnel, take filter Liquid, adds the extremely layering of a small amount of water in filtrate, takes off layer aqueous phase and extracts three times with ether and ethyl acetate respectively, and steaming is revolved in water intaking mutually And be dried, obtain faint yellow solid, i.e. tyrosine dithiocarbamate (TYRDTC)；Concrete synthetic route is:

b.^99mThe preparation of TcO-TYRDTC:

Appropriate Na is added in GH medicine box^99mTcO₄(37～740MBq), shake up rear room temperature and place 15 minutes, i.e. obtain^99mTcO-GH.1～2mg TYRDTC is joined^99mIn TcO-GH solution, boiling water bath heats 30 minutes, i.e. obtains institute of the present invention State^99mTcO_-TYRDTC complex.

^99mThe preparation employing ligand exchange reaction of TcO-TYRDTC:

Synthetic route is as follows:

^99mTcO₄ ^-+SnCl₂: 2H₂O+GH→^99mTcO-GH

^99mTcO_-GH+TYRDTC→^99mTcO_-TYRDTC。

Above-mentioned chemical synthesis reagent is all commercial goods, and wide material sources are easy to get.

Synthesized by said method^99mTcO-TYRDTC is a kind of novel tumor developer, and it prepares simplicity, activation Learn purity more than 90%.Tumor-bearing mice vivo biodistribution distribution experiments result shows:^99mTcO_-TYRDTC absorbs height in tumour, stagnant Staying the time long, tumour/muscle ratio is good, can become a kind of novel tumor developer.

Will^99mTcO-TYRDTC and¹⁸F-FDG compares at lotus S180 mice with tumor vivo biodistribution distributed data, the results are shown in Table 1

Table 1:^99mTcO-TYRDTC and¹⁸F-FDG is lotus S180 mice with tumor vivo biodistribution distribution (%ID/g) after injection 4h

Result above shows, although^99mThe tumour of TcO-TYRDTC/blood ratio to be less than¹⁸F-FDG, but taking the photograph in tumour Value is higher than¹⁸F-FDG, has reached 10.09 ± 0.87%ID/g, and tumour/muscle ratio is far above simultaneously¹⁸F-FDG, Ke Yizuo For novel tumor developer popularization and application.

Experiment shows,^99mThe performance of TcO-TYRDTC is as follows:

1.^99mThe chromatography of TcO-TYRDTC is identified:

Thin-layer chromatography chromatogram (TLC) is identified: with polyamide film as support, respectively with physiological saline and acetonitrile conduct Solvent, the tomographic results of mensuration is shown in Table 2.

Table 2: the tomographic results (R of each component_fResult)

The radiochemical purity of the label measured by above-mentioned chromatography is identified is more than 90%.

2.^99mThe mensuration of the lipid of TcO-TYRDTC

Take the phosphate buffer (0.025mol/L) of 1.0mL pH=7.4 in 10mL centrifuge tube, at centrifuge tube Middle addition 1.0mL n-octyl alcohol and 0.01mL^99mTcO-TYRDTC complex solution, covers stopper, fully shakes up, centrifugal 5min (14000r/min).From organic phase and aqueous phase, take out 0.1mL the most respectively, measure the radiocounting of two-phase, and calculate it Lipid P (radioactive activity of the radioactive activity/aqueous phase of P=organic phase), in triplicate, records logP=- 2.36+0.03, explanation^99mTcO-TYRDTC is hydroaropic substance.

3.^99mThe vitro stability of TcO-TYRDTC and serum stability measure

By marked^99mTcO-TYRDTC at room temperature places different time (1,2,4,6 hours), and to measure it afterwards activation Learn purity, test result indicate that^99mAfter TcO-TYRDTC at room temperature places 6 hours, radiochemical purity is more than 90%, explanation^99mTcO-TYRDTC at room temperature vitro stability is good, meets the needs of clinical practice.By marked^99mTcO-TYRDTC adds Entering in 0.1% haemocyanin, after placing 4h at 37 DEG C, radiochemical purity is more than 90%, explanation^99mTcO-TYRDTC is at blood Have good stability in Qing.4.^{Ovum m}TcO-TYRDTC biodistribution experiments in bearing mouse model:

Tail vein injection 0.10mL by the kunming mice (female, body weight about 18-20g) of lotus S180 sarcoma model^99mTcO-TYRDTC solution (about 7.4 × 10⁵Bq), the sacrificed by decapitation when 0.5h, 2h, 4h respectively after injection.Take its heart, liver, lung, Related organization and the organs such as kidney, spleen, muscle, bone, blood, tumour, weigh after cleaning respectively, and on FM-2000 type technetium analyzer Surveying its radiocounting, time each, the tumor-bearing mice number of item is 3.Calculate every gram of percentage injection dosage (%ID/ of each tissue g).The results are shown in Table 3.

Table 3:^99mTcO-TYRDTC is at lotus S180 sarcoma Biodistribution in mice (n=3) %ID/g

5.⁹⁹MTcO-TYRDTC tumor-bearing mice Animal imaging is tested

Tail vein injection 0.10mL by the kunming mice (female, body weight about 18-20g) of lotus S180 sarcoma model^99mTcO-TYRDTC solution (about 3.7 × 10⁷Bq), after injection, prostrate its four limbs of fixing of 4h carry out SPECT imaging (imaging SPECT Instrument is Eplus-166, uses the sensitivity of 38cps/MBq to be scanned, and the static scanning time is 10min).SPECT images Result shows^99mTcO-TYRDTC has obvious concentration in tumour stove.

Illustrated by the above detection data implemented,^99mTcO-TYRDTC complex can be as a kind of novel tumor imaging Agent carries out popularization and application.

Detailed description of the invention:

Below by the embodiment detailed description present invention:

A kind of^99mTcO-TYRDTC complex complex preparation method, it represents formula and is^99mTcO-TYRDTC, its part TYRDTC structural formula is as follows:

Its preparation method is as follows:

A. the synthesis of part TYRDTC:

1025mg (5.70mmol) L-tyrosine and 456mg (11.4mmol) NaOH it is separately added in reaction bulb, with 10mL absolute methanol, as solvent, ice-water bath stirring and dissolving, is added dropwise over 5mL dissolved with 1mL (17.4mmol) C8₂Ether molten Liquid, reacts 3 hours after being added dropwise to complete in ice-water bath, takes filtrate, add a small amount of water extremely in filtrate after filtering with sand core funnel Layering, takes off layer aqueous phase and extracts three times with ether and ethyl acetate respectively, and water intaking rotation mutually is steamed and is dried, and obtains faint yellow solid 0.81g, productivity is the tyrosine dithiocarbamate (TYRDTC) of 48%, records its specific rotatory power for [α]_D ²⁵(H₂=+67.03 ° O)；

Its concrete synthetic route is:

Infrared analysis qualification result IR (KBr)/cm^-1: 3432.60 (OH), 1631.14 (C=O), 1111.56 (C=S).

Hydrogen spectrum qualification result¹H-NMR(D₂O) δ: 7.00-7.02 (d, 2H), 6.71-6.73 (d, 2H), 4.75-4.77 (dd, 1H), 3.06-3.11 (dd, 1H), 2.84-2.89 (dd, 1H).

Carbon spectrum qualification result¹³C-NMR(D₂O) δ: 210.45 (N-CS₂), 177.94 (COONa), 154.36 (COH), 130.68(CHCCH), 129.22 (CCH₂), 115.45 (CHCOHCH), 63.79 (CHCOONa), 36.38 (CH₂CH).

C₁₀H₉NNa₂O₃S₂·1.5H₂O elementary analysis result theoretical value: C, 36.59；N, 4.27；H, 3.35. measured value For: C, 36.98；N, 4.33；H, 3.50.

b.^99mTcO_-The synthesis of TYRDTC

Prepared by employing ligand exchange method^99mTcO-TYRDTC, its synthetic route is:

^99mTcO₄ ^-+SnCl₂.2H₂O+GH→^99mTcO-GH

^99mTcO-GH+TYRDTC→^99mTcO_-TYRDTC

Appropriate Na is added in GH medicine box (gluceptate medicine box)^99mTcO₄(37～740MBq), shake up rear room temperature and place 15 minutes, i.e. obtain^99mTcO-GH.1～2mg TYRDTC is joined^99mIn TcO-GH solution, boiling water bath heating 30 minutes, i.e. Obtain of the present invention^99mTcO-TYRDTC complex.

Claims

1. one kind^99mThe preparation method of TcO-TYRDTC complex, it represents formula and is^99mTcO-TYRDTC, described part TYRDTC structural formula is as follows:

Its preparation method is as follows:

A. the synthesis of part TYRDTC:

Being separately added into a certain amount of TYR, NaOH in reaction bulb, using absolute methanol as solvent, ice-water bath stirring is molten Solve, instill the CS being dissolved in ether₂, react 3 hours in ice-water bath after being added dropwise to complete, after filtering with sand core funnel, take filtrate, Adding the extremely layering of a small amount of water in filtrate, take off layer aqueous phase and extract three times with ether and ethyl acetate respectively, water intaking is revolved mutually and is steamed also It is dried, obtains faint yellow solid, i.e. TYRDTC；Concrete synthetic route is:

b.^99mThe preparation of TcO-TYRDTC:

The Na of 37～740MBq is added in GH medicine box^99mTcO₄, shake up rear room temperature and place 15 minutes, i.e. obtain^99mTcO-GH；By 1 ～2mg TYRDTC joins^99mIn TcO-GH solution, boiling water bath heats 30 minutes, i.e. obtains described^99mTcO-TYRDTC joins Compound:

^99mThe preparation employing ligand exchange reaction of TcO-TYRDTC:

Synthetic route is as follows:

^99mTcO₄ ^-+SnCl₂·2H₂O+GH→^99mTcO-GH

^99mTcO-GH+TYRDTC→^99mTcO-TYRDTC。

The most as claimed in claim 1^99mThe preparation method of TcO-TYRDTC complex is used for preparing tumor imaging at the field of nuclear medicine Application in agent.