CN103304493A

CN103304493A - 2-fluoro-aniline quinazoline tumour positron imaging agents as well as preparation method and application thereof

Info

Publication number: CN103304493A
Application number: CN2013102564748A
Authority: CN
Inventors: 刘振锋; 董孟杰; 王国林; 张倩; 晋建文; 赵葵
Original assignee: Zhejiang University ZJU
Current assignee: Zhejiang University ZJU
Priority date: 2013-06-24
Filing date: 2013-06-24
Publication date: 2013-09-18
Anticipated expiration: 2033-06-24
Also published as: CN103304493B

Abstract

The invention provides a 2-fluoro-aniline quinazoline compound labelled by F-18, wherein 18 F is produced by a circular accelerator via 18O(pn)18F nuclear reaction and automatically synthesised by a radioactive synthesis module, and can also be produced by the existing domestic F-18 multifunctional synthesis device after process reformation. The 2-fluoro-aniline quinazoline compound provided by the invention is of an aniline quinazoline structure substituted by 2-bit positron nuclide fluorine-18, and can be modified on 6-bit, 7-bit and benzene ring connected with an amino group. The invention provides novel tumour positron imaging agents, wherein compared with 18F-fluoro-deoxyglucose (18FDG), the imaging agents are specific, and capable of identifying the tumours highly expressed by an epidermal growth factor receptor (EGFR). The preparation method is reasonable in design, simple in labelling method, capable of realizing automatic production, and suitable for application. The structural general formula of the 2-fluoro-aniline quinazoline compound is defined in the specification.

Description

2-Fluoroaniline quinazoline ditosylate salt tumour pet imaging agent and preparation and application

Technical field

The invention belongs to field of medicaments, relate to a kind of aniline quinazoline compound, relate in particular to 2-Fluoroaniline quinazoline compounds of F-18 mark and preparation method thereof, and the application in preparation tumour positron emission fault (PET) developer.

Background technology

PET is as present state-of-the-art Medical Imaging Technology, can realize high-resolution video picture is carried out in cellular metabolism and function, from molecular level, physiology, the biological process of human body are carried out nothing wound, three-dimensional, dynamic studies, PET is applied to the diagnosis that tumour comprises tumour, good pernicious discriminating, malignant tumour by stages, early diagnosis and the discriminating of somatotype, Preventive, the observation of the selection for the treatment of plan and the monitoring of chemical therapeutic effect and tumour change procedure and more after the detection of situation.The PET inspection is different from other inspection, and it depends on positron medicine (PET medicine), the purpose that relies on the positron medicine specificity to concentrate and reach diagnosis and estimate in target organ.The topmost positron medicine of using during current PE T checks is ¹⁸The F-fluorodeoxyglucose ( ¹⁸FDG), ¹⁸FDG is as a kind of tumour metabolism class medicine, malignant tumour diagnose with differential diagnosis in brought into play huge effect, still ¹⁸FDG as not distinguishing inflammation and tumour, much is reported as false positive or false negative as a kind of relatively non-specific developer, about the diagnosis of tumour and research also need to develop novel, have more specific developer.The aniline quinazoline compounds is to be that a class is of many uses so far, has the important heterogeneous ring compound than high biological activity, main manifestations is active to the inhibition of EGFR or its Tyrosylprotein kinase (EGFR-TK), vascular endothelial growth factor receptor (VEGFR), platelet derived growth factor receptor (PDGFR), trk C (NGFR) and other a plurality of action target spots, thereby bring into play the multiple pharmacological effect such as anticancer, antibiotic, antiviral, thereby it becomes the focus of the world of medicine and region of chemistry research.

Summary of the invention

The purpose of this invention is to provide a kind of 2-Fluoroaniline quinazoline compound of F-18 mark, structural formula is as follows:

Wherein:

Structural formula be characterized as the aniline quinazoline structure that 2 positron radionuclide fluoro-18 replace, can be at 6,7, and with phenyl ring that amino links to each other on the structure of modifying.

Particular compound relates to:

。

Another object of the present invention provides the preparation method of the 2-Fluoroaniline quinazoline compound of F-18 mark, by the following method 1 or method 2 realize:

Method 1: use magnetic resonance acceleator and use bombardment ¹⁸O water, by ¹⁸O (p n) ¹⁸The F nuclear reaction is produced and is obtained ¹⁸The F ion is with 1mL mixing solutions (mixing solutions: 12.5 mg 4,7,13,16,21,24-six oxygen-1,10-diaza-bicyclo [8.8.8] hexacosane (K ₂₂₂) add 2.5mg K ₂CO ₃Be dissolved in 0.1mL water and 0.9mL second eyeball) will ¹⁸F drip washing is in reaction tubes, evaporate to dryness, add 2mL second eyeball, evaporate to dryness again, 2 halogens that add 5mg replace organic solution (methyl-sulphoxide, the N of 4-aniline quinazoline precursors, dinethylformamide, second eyeballs etc.), 100-140 ℃ of reaction 10min obtains the 4-aniline quinazoline compounds that corresponding 2 fluoro-18 replace; Separate to obtain product with HPLC through the C-18 pillar, separator column is the C-18 post of preparation type or half preparation type, and moving phase is the leacheate of second eyeball and water or the leacheate of ethanol and water.Reaction formula:

。

Method 2: use magnetic resonance acceleator and use bombardment ¹⁸O water, by ¹⁸O (p n) ¹⁸The F nuclear reaction is produced and is obtained ¹⁸The F ion is with 1mL solution (mixing solutions: 12.5 mg 4,7,13,16,21,24-six oxygen-1,10-diaza-bicyclo [8.8.8] hexacosane (K ₂₂₂) add 2.5mg K ₂CO ₃Be dissolved in 0.1mL water and 0.9mL second eyeball) will ¹⁸F drip washing is in reaction tubes, evaporate to dryness adds 2mL second eyeball, again evaporate to dryness, 2 halogens of 4-N-tertiary butyl carbonyl that add 5mg replace the organic solution (methyl-sulphoxide of 4-aniline quinazoline precursor, DMF, second eyeball etc.), 100-140 ℃ of reaction 10min, add hydrochloric acid, slough protection, obtain the 4-aniline quinazoline compounds that corresponding 2 fluoro-18 replace; Separate to obtain product with HPLC through the C-18 pillar, separator column is the C-18 post of preparation type or half preparation type, and moving phase is the leacheate of second eyeball and water or the leacheate of ethanol and water.Reaction formula:

。

Halogen described in the reaction is chlorine, bromine or iodine.

The 2-Fluoroaniline quinazoline compound that the 3rd purpose of the present invention provides the F-18 mark prepares the application in the PET developer, experimental results show that the 2-Fluoroaniline quinazoline compound of F-18 mark can be used for the especially PET video picture of Urogastron (EGFR) high expression level tumour of tumour.Be a class new at the tumour pet imaging agent.

The present invention is passed through by magnetic resonance acceleator ¹⁸O (p n) ¹⁸The F nuclear reaction is produced ¹⁸F, it is synthetic that such developer can carry out automatization by the radiation synthesis module.The present invention can also can pass through the multi-functional synthesizer of existing domestic F-18 (Beijing Paite Co., Ltd.) by manually synthetic preparation, produces after workflow reengineering, and production process is simple, also passes through other commercial full-automatic synthesizer.

The invention provides the novel pet imaging agent of a class (2-Fluoroaniline quinazoline ditosylate salt pet imaging agent), with and preparation method thereof with application, with ¹⁸The F-fluorodeoxyglucose ( ¹⁸FDG) compare, such developer has specificity, can identify those Urogastrons (EGFR) high expression level tumour; In the preparation method, 2 fluoro-18 mark aniline quinazoline compounds, marking method is simple, and can realize automatic production, can satisfy the needs of scientific research and clinical experiment.

Description of drawings

Fig. 1 is the automatization synthesizer schematic flow sheet of 2 fluorine marks of 4-aniline quinazoline class.

Fig. 2 is 2- ¹⁸F-6,7-dimethoxyquinazoline-4-aniline radioactivity HPLC collection of illustrative plates.

Fig. 3 is 2- ¹⁸F-6, the outer HPLC collection of illustrative plates of 7-dimethoxyquinazoline-4-mauvein (e).

Fig. 4 is 2- ¹⁸F-6,7-dimethoxyquinazoline-4-(3 '-bromine) the radioactivity HPLC spectrogram of amine.

Fig. 5 is 2- ¹⁸F-6,7-dimethoxyquinazoline-4-(3 '-bromine) the ultraviolet HPLC spectrogram of amine.

Fig. 6 is 2- ¹⁸F-6,7-dimethoxyquinazoline-4-(3 '-alkynyl) the radioactivity HPLC spectrogram of aniline.

Fig. 7 is 2- ¹⁸F-6,7-dimethoxyquinazoline-4-(3 '-alkynyl) the ultraviolet HPLC spectrogram of aniline.

Fig. 8 is 2- ¹⁸The bio distribution of F-PD153035.

Fig. 9 is the 2-of PC9 mice with tumor ¹⁸The video picture of F-PD153035 small animal position emission tomography (PET).

Embodiment

The present invention is further described in conjunction with the accompanying drawings and embodiments.But the present invention is not limited.

The C18 pillar is Sep-pak Plus C18 among the embodiment, Sep-pak Plus tC18, Sep-Pak Light C18 pillar, Oasisis HLB Plus pillar [being U.S. Waters company product] or also can make the C18 pillar by oneself.

Separator column is the C-18 post of preparation type or half preparation type, and moving phase is the leacheate of second eyeball and water or the leacheate of ethanol and water, and separation obtains target product to reaction mixture through HPLC.

Embodiment 1 2- ¹⁸F-6, the radiosynthesis of 7-dimethoxyquinazoline-4-aniline

Referring to Fig. 1, use magnetic resonance acceleator and use bombardment ¹⁸O water, by ¹⁸O (p n) ¹⁸The F nuclear reaction is produced and is obtained 100mCi ¹⁸The F ion will speed up that device produces ¹⁸F is caught by the QMA post;

⑴ 1mL solution (the 12.5 mg K in the headpin ₂₂₂Add 2.5mg K ₂CO ₃Be dissolved in the mixing solutions of 0.1mL water and 0.9mL second eyeball) will ¹⁸F drip washing is in reaction tubes;

⑵ dry up the solution in the reaction tubes in 116 ℃ of nitrogen;

⑶ the dry second eyeball of 2mL in No. 2 bottles of adding dries up in 116 ℃ of nitrogen in the reaction tubes;

⑷ 5mg 2-chloro-6 in No. 3 bottles of adding in the reaction tubes, the dry DMSO of 7-dimethoxyquinazoline-4-aniline precursor and 0.8mL is in 140 ℃ of reaction 10min;

⑸ minute three adding kingpin 30mL water in the reaction tubes, by the C-18 post between V7 and the V10 valve, marked product is trapped on the C-18 post, and the impurity such as precursor negative ion are transferred in the waste liquid bottle by the V10 T-valve with mixing liquid;

⑹ add the 2mL ethanolic soln of No. 6 bottles in the reaction tubes, the marker on the C-18 post is washed, thick product application HPLC separates, and separation condition is 50% ethanol: the moving phase of water, the C-18 semipreparative column obtains product.

Embodiment 2 2- ¹⁸F-6, the radiosynthesis of 7-dimethoxyquinazoline-4-aniline

Use magnetic resonance acceleator and use bombardment ¹⁸O water, by ¹⁸O (p n) ¹⁸The F nuclear reaction is produced and is obtained 100mCi ¹⁸The F ion will speed up that device produces ¹⁸F is caught by the QMA post;

⑵ dry up the solution in the reaction tubes in 116 ℃ of nitrogen;

⑷ 5mg 2-bromo-6 in No. 3 bottles of adding in the reaction tubes, the dry DMF of 7-dimethoxyquinazoline-4-N-tertiary butyl carbonyl-aniline and 0.8mL is in 140 ℃ of reaction 10min;

⑸ the 2mL hydrochloric acid soln that adds No. 6 bottles in the reaction tubes, hydrolysis 15min

⑹ minute three adding kingpin 30mL water in the reaction tubes, by the C-18 post between V7 and the V10 valve, marked product is trapped on the C-18 post, and the impurity such as precursor negative ion are transferred in the waste liquid bottle by the V10 T-valve with mixing liquid;

Add the 2mL ethanolic soln of No. 4 bottles in the reaction tubes, the marker on the C-18 post is washed, thick product application HPLC separates, and separation condition is 50% ethanol: the moving phase of water, the C-18 semipreparative column obtains product.

Embodiment 3 2- ¹⁸F-6, the quality control of 7-dimethoxyquinazoline-4-aniline

The HPLC collection of illustrative plates is referring to Fig. 2,3, the Quality Control condition is the 50% second eyeball aqueous solution, the post of C-18(4.5mm * 250mm), ultraviolet 254nm and radioactivity seeker, uv-absorbing is consistent with the radioactivity absorption recklessly for the display standard product as a result, be same substance, generated time 45min does not proofread and correct synthetic yield 20-30%, radiochemical purity〉98%.The standard substance spectrogram is: ESI-MS (m/z, %): 300 ([M+H] ⁺, 100), ¹H NMR (CDCl3,400MHz): 7.69 (2H, d, J=7.93,4 phenyl ring 2 '-H, 6 '-H); 7.45 (1H, br.s ,-NH); 7.42 (2H, dd, J ₁=7.53Hz, J ₂=8.32Hz, 4 phenyl ring 3 '-H, 5 '-H); 7.20 (1H, dd, J ₁=7.14Hz, J ₂=6.34Hz, 4 phenyl ring 4 '-H); (7.15 1H, s, 8-H); (7.04 1H, s, 5-H); 4.00 (6H, s, 2-OCH ₃).

Embodiment 4 2- ¹⁸F-6,7-dimethoxyquinazoline-4-(3 '-bromine)-radiation of aniline is synthetic

⑵ dry up the solution in the reaction tubes in 116 ℃ of nitrogen;

⑷ 5mg 2-chloro-6 in No. 3 bottles of adding in the reaction tubes, 7-dimethoxyquinazoline-4-(3 '-bromine)-aniline precursor and the dry DMSO of 0.8mL, in 140 ℃ of reaction 10min;

⑹ the 2mL ethanolic soln that adds No. 6 bottles in the reaction tubes,, the marker on the C-18 post is washed, thick product application HPLC separates, and separation condition is 50% ethanol: the moving phase of water, the C-18 semipreparative column obtains product.

Embodiment 5 2- ¹⁸F-6,7-dimethoxyquinazoline-4-(3 '-bromine)-radiation of aniline is synthetic

⑵ dry up the solution in the reaction tubes in 116 ℃ of nitrogen;

⑷ 5mg 2-iodo-6 in No. 3 bottles of adding in the reaction tubes, the dry DMF of 7-dimethoxyquinazoline-4-N-tertiary butyl carbonyl-(3 '-bromine)-aniline precursor and 0.8mL is in 140 ℃ of reaction 10min;

⑸ the 2mL hydrochloric acid soln that adds No. 6 bottles in the reaction tubes, hydrolysis 15min;

Embodiment 6 2- ¹⁸F-6,7-dimethoxyquinazoline-4-(3 '-bromine) quality control of amine

The HPLC collection of illustrative plates is seen Fig. 4,5, the Quality Control condition is the 60% second eyeball aqueous solution, the post of C-18(4.5mm * 250mm), ultraviolet 254nm and radioactivity seeker, display standard product uv-absorbing is consistent with the radioactivity absorption as a result, is same substance, generated time 45min, do not proofread and correct synthetic yield 10-20%, radiochemical purity〉98%.The standard substance spectrogram is: ESI-MSm/z (%): 378 ([M+1], 100), and 380 ([M+3], 100), ¹H NMR (CDCl ₃, 400 Hz) δ: 7.88 (1H, t, J=1.76 Hz, 2 '-H), 7.71 (1H, dt, J ₁=7.82 Hz, J ₂=1.76 Hz, 6 '-H), 7.32 (1H, br s ,-NH-), 7.30 (1H, dd, J ₁=7.82 Hz, J ₂=7.44 Hz, 5 '-H), 7.26 (1H, d, J=7.44 Hz, 4 '-H), 7.16 (1H, s, 8-H), 6.99 (1H, s, 5-H), 4.02 (3H, s ,-OCH ₃), 4.01 (3H, s ,-OCH ₃).

Embodiment 7 2- ¹⁸F-6,7-dimethoxyquinazoline-4-(3 '-alkynyl) aniline is synthetic

⑵ dry up the solution in the reaction tubes in 116 ℃ of nitrogen;

⑷ 5mg 2-chloro-6 in No. 3 bottles of adding in the reaction tubes, 7-dimethoxyquinazoline-4-(3 '-alkynyl)-aniline precursor and the dry DMSO of 0.8mL, in 140 ℃ of reaction 10min;

Embodiment 8 2- ¹⁸F-6,7-dimethoxyquinazoline-4-(3 '-alkynyl) aniline is synthetic

⑵ dry up the solution in the reaction tubes in 116 ℃ of nitrogen;

⑷ 5mg 2-iodo-6 in No. 3 bottles of adding in the reaction tubes, the dry acetonitrile of 7-dimethoxyquinazoline-4-N-tertiary butyl carbonyl-(3 '-alkynyl)-aniline precursor and 0.8mL is in 140 ℃ of reaction 10min;

(7) the 2mL ethanolic soln of No. 4 bottles of adding in the reaction tubes washes the marker on the C-18 post get off, and thick product application HPLC separates, and separation condition is 50% ethanol: the moving phase of water, the C-18 semipreparative column obtains product.

Embodiment 9 2- ¹⁸F-6,7-dimethoxyquinazoline-4-(3 '-alkynyl) aniline is manually synthetic

Use magnetic resonance acceleator and use bombardment ¹⁸O water, by ¹⁸O (p n) ¹⁸The F nuclear reaction is produced and is obtained 20mCi ¹⁸The F ion will speed up that device produces ¹⁸F is caught by the QMA post; 1mL solution (12.5 mg K ₂₂₂Add 2.5mg K ₂CO ₃Be dissolved in the mixing solutions of 0.1mL water and 0.9mL second eyeball) will ¹⁸F drip washing is in the 10mL glass reaction tube; Solution in the reaction tubes is dried up in 116 ℃ of nitrogen; Add the dry second eyeball of 2mL in the reaction tubes, in 116 ℃ of nitrogen evaporates to dryness, add 2-iodo-6,7-dimethoxyquinazoline-4-(3 '-alkynyl in the reaction tubes)-the aniline precursor, and the dry DMSO of 0.8mL, in 140 ℃ of reaction 10min; Adding 10mL water joins in the reaction tubes, manual transfer is to the C-18 pillar, continue again the pillar with 20mL water flushing C-18, use the 2mL ethanolic soln, compound with the thick step purification on the C-18 post, wash and use the HPLC separation, separation condition is 50% ethanol: the moving phase of water, the C-18 semipreparative column obtains product.

Embodiment 10 2- ¹⁸F-6,7-dimethoxyquinazoline-4-(3 '-alkynyl) quality control of aniline

The HPLC collection of illustrative plates is referring to Fig. 6,7, the Quality Control condition is the 65% second eyeball aqueous solution, the post of C-18(4.5mm * 250mm), ultraviolet 254nm and radioactivity seeker, uv-absorbing is consistent with the radioactivity absorption recklessly for the display standard product as a result, is same substance, generated time 45min, do not proofread and correct synthetic yield 10-20%, radiochemical purity〉98%.The standard substance spectrogram is: ESI-MSm/z (%): 324 ([M+1], 100), ¹H NMR (CDCl ₃, 400 Hz) δ: 7.78 (1H, d, J=8.21 Hz, 6 '-H), 7.75 (1H, s, 2 '-H), 7.40 (1H, br s ,-NH-), 7.36 (1H, dd, J ₁=7.82 Hz, J ₂=7.83 Hz, 5 '-H), 7.29 (1H, d, J=7.83 Hz, 4 '-H), 7.14 (1H, s, 8-H), 7.01 (1H, s, 5-H), 4.01 (3H, s ,-OCH ₃), 3.99 (3H, s ,-OCH ₃).

Embodiment 11 2- ¹⁸The stability experiment of F-PD153035

2- ¹⁸The stability experiment of F-PD153035 shows, observes 8 hours in 37 degrees centigrade calf serum, and radiochemical purity still can reach more than 98%, has good stability.Bio distribution at normal Kunming mouse is seen Fig. 8, shows that drug main will pass through the liver sausage metabolism, and the blood removing speed is fast, and radioactive uptake is not high in the bone, and not defluorinate in the prompting medicine body is stable in vivo.

Embodiment 12

2- ¹⁸The PC9 mice with tumor small animal position emission tomography (PET) video picture (Fig. 9) of F-PD153035 shows that tumor locus has radioactive uptake.This explanation 2-Fluoroaniline quinazoline compounds is expressed high tumour to which EGFR and can be carried out PET tumor imaging, and such medicine can be used as the new tumor developer of a class.

Claims

1. the 2-Fluoroaniline quinazoline compound of a F-18 mark, structural formula is as follows:

。

2. the 2-Fluoroaniline quinazoline compound of a kind of F-18 mark according to claim 1, it is characterized in that, described compound is the aniline quinazoline structure that 2 positron radionuclide fluoro-18 replace, at 6,7, and with phenyl ring that amino links to each other on the structure of modifying, be specifically related to following compound:

。

3. the preparation method of the 2-Fluoroaniline quinazoline compound of F-18 mark according to claim 1 is characterized in that, realizes by following steps:

Use magnetic resonance acceleator and use bombardment ¹⁸O water, by ¹⁸O (p n) ¹⁸The F nuclear reaction is produced and is obtained ¹⁸The F ion will with the 1mL mixing solutions ¹⁸F drip washing is in reaction tubes, and evaporate to dryness adds 2mL second eyeball, evaporate to dryness again, and 2 halogens that add 5mg replace the organic solution of 4-aniline quinazoline precursors, and 100-140 ℃ of reaction 10min obtains the 4-aniline quinazoline compounds that 2 fluoro-18 replace; Separate with HPLC through the C-18 pillar and to obtain product, separator column is the C-18 post of preparation type or half preparation type, moving phase is the leacheate of second eyeball and water or the leacheate of ethanol and water, wherein mixing solutions forms: 12.5 mg 4,7,13,16,21,24-, six oxygen-1,10-diaza-bicyclo [8.8.8] hexacosane adds 2.5mg K ₂CO ₃Be dissolved in 0.1mL water and 0.9mL second eyeball, reaction formula:

。

4. the preparation method of the 2-Fluoroaniline quinazoline compound of F-18 mark according to claim 1 is characterized in that, realizes by following steps: use magnetic resonance acceleator and use bombardment ¹⁸O water, by ¹⁸O (p n) ¹⁸The F nuclear reaction is produced and is obtained ¹⁸The F ion will with 1mL solution ¹⁸F drip washing is in reaction tubes, and evaporate to dryness adds 2mL second eyeball, evaporate to dryness again, 2 halogens of 4-N-tertiary butyl carbonyl that add 5mg replace the organic solution of 4-aniline quinazoline precursor, 100-140 ℃ of reaction 10min, add hydrochloric acid, slough protection, obtain the 4-aniline quinazoline compounds that 2 fluoro-18 replace; Separate to obtain product with HPLC through the C-18 pillar, separator column is the C-18 post of preparation type or half preparation type, and moving phase is the leacheate of second eyeball and water or the leacheate of ethanol and water;

Reaction formula:

。

5. according to claim 3 or the preparation method of the 2-Fluoroaniline quinazoline compound of 4 described F-18 marks, it is characterized in that, wherein said halogen is chlorine, bromine or iodine.

6. according to claim 3 or the preparation method of the 2-Fluoroaniline quinazoline compound of 4 described F-18 marks, it is characterized in that, organic solution is selected methyl-sulphoxide, DMF or second eyeball.

7. according to claim 3 or the preparation method of the 2-Fluoroaniline quinazoline compound of 4 described F-18 marks, it is characterized in that, ¹⁸F is passed through by magnetic resonance acceleator ¹⁸O (p n) ¹⁸The F nuclear reaction is produced and is obtained.

8. the application of the 2-Fluoroaniline quinazoline compound of a kind of F-18 mark according to claim 1 in preparation PET developer.