CN109568606A - Target TSPO magnetic resonance imaging contrast and preparation method thereof - Google Patents
Target TSPO magnetic resonance imaging contrast and preparation method thereof Download PDFInfo
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- A61K49/06—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations
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Abstract
The present invention relates to a kind of targeting TSPO magnetic resonance imaging contrasts and preparation method thereof, to solve the problems, such as that it is low that the prior art images spatial resolution.Be made of the following steps: the preparation of step (1) CB86-DTPAA: 1) being added DMSO, DTPAA to the bottle of tinfoil paper paper bag, is separately slowly added to CB86 by several times, and stirring while adding；2) triethylamine is added, sealing in magnetic heating stirrer in being protected from light；3) it is put into refrigerator to be lyophilized overnight, next day takes out to be placed on freeze dryer and be dehydrated, and pure water is redissolved in after being lyophilized, and continues freeze-drying overnight, repeated multiple times to obtain powder；4.) it after purification, is evaporated, obtains CB86-DTPAA powder；The CB86-DTPAA connection gadolinium of step (2) after purification: 1) it into the CB86-DTPAA powder of above-mentioned preparation is added gadolinium trichloride, is dissolved in PBS, is stirred overnight；2) after purified, revolving obtains final product CB86-DTPAA-Gd.It can be used for the early diagnosis of periphery chronic inflammation disease and curative effect evaluation, have imaging spatial resolution high, soft tissue resolution is good, and "dead" advantage.
Description
Technical field
The present invention relates to a kind of magnetic resonance contrast agent, more particularly to a kind of targeting TSPO magnetic resonance imaging contrast and its
Preparation method.
Background technique
Magnetic resonance imaging (MRI) has spatial resolution high, and soft tissue resolution is good, clearly displays soft group of perilesional
Knit knot, and it is reproducible the advantages that, it has also become one of clinical important detection methods.Current clinically common gadolinium contrast agent is because of nothing
The defects of targeting, there is poor specificities, therefore develop the hot spot that targeted MRI contrast agent has been the area research.Periphery
High expression level is presented in type benzodiazepine receptors transport protein (TSPO) in inflammatory cell (activated macrophage).TSPO expression
Amount can reflect macrophage accumulation and activation degree to a certain extent, and then reflect the process of course inflammatory activity.Target at present
To the molecular image probe of TSPO be mostly radionuclide (18F or99mTc) mark row PET or SPECT imaging, PET or
The defects of that there is spatial resolutions is low for SPECT imaging, soft tissue resolution difference and radioactivity, therefore develop targeting TSPO magnetic
Resonance image-forming contrast agent has extremely important meaning.
Summary of the invention
Present invention aims to overcome that the drawbacks described above of the prior art, provides a kind of imaging spatial resolution height, soft tissue
The targeting TSPO magnetic resonance imaging contrast of good resolution, the object of the invention also reside in offer preparation method.
To achieve the above object, the present invention targets TSPO magnetic resonance imaging contrast, its special feature is that by following step
CB86-DTPAA-Gd made of rapid.
The preparation of step (1) CB86-DTPAA: 1) to being placed in magnetic heating stirrer and encase body with masking foil
Eggplant-shape bottle DMSO is added, and DTPAA is added, CB86 is separately slowly added to eggplant-shape bottle by several times, and want stirring while adding, it is necessary
Shi Jinhang ultrasonic dissolution assisting；2) triethylamine is added to eggplant-shape bottle, packs tightly bottleneck with sealed membrane or is stoppered with bottle stopper, and heated in magnetic force
It is protected from light on blender；3) it being lyophilized: eggplant-shape bottle being first put into refrigerator freeze-drying overnight, next day takes out to be placed on freeze dryer and be dehydrated,
It is redissolved in pure water after being lyophilized, continues freeze-drying overnight, repeated multiple times obtained powder；4.) it purifies: after activation C18 pillar, (1)
Powder in step 3) is dissolved in NaOH solution, ultrasonic dissolution assisting extracts every time, the C18 pillar after squeezing into activation, after rushed with air
Once；(2) NA is taken2CO3Solution, the C18 pillar of step before rinsing；(3) pure water rinsing once before step C18 pillar, then with empty
Gas punching is primary, discards filtrate；(4) dehydrated alcohol, the C18 pillar of step, air rush again before rinsing, and collect ethanol liquid in centrifugation
Pipe；(5) penicillin bottle is added in the ethanol rinse liquid being collected into batches, is evaporated with revolving instrument, CB86-DTPAA powder is obtained
End.
The CB86- DTPAA connection gadolinium of step (2) after purification: 1) it into the CB86-DTPAA powder of above-mentioned preparation is added
Gadolinium trichloride is dissolved in PBS, is stirred overnight；2) after purified, revolving obtains final product CB86-DTPAA-Gd.It is purified to be
It is purified through above-mentioned steps (one) purification process.High with imaging spatial resolution, soft tissue resolution is good, and "dead" excellent
Point.
As optimization, the step 1) of the CB86-DTPAA preparation is: standby 50ml eggplant-shape bottle one, and is encased with masking foil
Body is placed in magnetic heating stirrer；5ml DMSO is added, and is protected from light lower addition 800mg DTPAA, separately weighs 200mg
CB86 is poured slowly into 50ml eggplant-shape bottle by several times, pays attention to slowly instilling, and wants to rotate in drop, it may be necessary to ultrasound
Hydrotropy.
The step 1) of the CB86- DTPAA connection gadolinium after purification is the CB86-DTPAA powder by above-mentioned preparation by 5:
Gadolinium trichloride is added in 1 ratio thereto, is dissolved in the PBS that PH is 7.4-8.0,1200rpm is stirred overnight.
As optimization, the step 2) of CB86-DTPAA preparation is: the triethylamine of 300ul being added to eggplant-shape bottle, uses sealed membrane
It packs tightly bottleneck or is stoppered with bottle stopper, in being protected from light -36h for 24 hours in magnetic heating stirrer.
As optimization, the step 3) of CB86-DTPAA preparation is freeze-drying: eggplant-shape bottle being first put into -80 DEG C of refrigerators and was lyophilized
Night, next day take out and are placed on freeze dryer and are dehydrated, and 20ml pure water is redissolved in after being lyophilized, and continue freeze-drying overnight, repeatedly 3 times powder
End.
As optimization, the activation C18 pillar in the step 4) of CB86-DTPAA preparation is: 10ml absolute alcohol is primary,
10ml water is primary, and air is primary, and moisture is gone out.
As optimization, (1) is the step of the step 4) of CB86-DTPAA preparation: the powder in step 3) is pressed
The ratio of 20mg:10ml is dissolved in the NaOH of 1mmol/l, and ultrasonic dissolution assisting extracts 5-10ml every time, the C18 pillar after squeezing into activation,
It is rushed afterwards with air primary.
As optimization, CB86-DTPAA preparation step 4) the step of (2) be: syringe takes the NA of 1mmol/l2CO3Solution
10ml, the C18 pillar of step before rinsing.
As optimization, (3) are the step of the step 4) of CB86-DTPAA preparation: pure water 10ml rinse it is primary before step
C18 pillar, then rushed once with air, discard filtrate.
As optimization, (4) are the step of the step 4) of CB86-DTPAA preparation: dehydrated alcohol 5ml, step before rinsing
C18 pillar, air rush again, collect ethanol liquid in 50ml centrifuge tube.
As optimization, purification process purifying of the step 2) through above-mentioned steps one in step (2) is purification step referring to step
Suddenly c, d, e in (one) obtains final product CB86-DTPAA-Gd after revolving.Specifically: being extracted in 50ml eggplant-shape bottle with syringe
CB86-DTPAA reacted with GdCl3 after solution 5ml, be injected into the C18 pillar of activation, then use 10ml pure water rinsing
C18 pillar is primary, then is rushed again once with air, discards filtrate, then then rinse C18 pillar with the dehydrated alcohol of 5ml, uses mould
Plain bottle collects dehydrated alcohol leacheate, is placed on revolving instrument and rotates, obtaining powder is final product CB86-DTPAA-Gd.
The preparation method of targeting TSPO magnetic resonance imaging contrast of the present invention, its special feature is that steps are as follows.
(1) preparation of CB86-DTPAA: 1) to the eggplant for being placed in magnetic heating stirrer and being encased with masking foil body
DMSO is added in shape bottle, and DTPAA is added, and CB86 is separately slowly added to eggplant-shape bottle by several times, and want stirring while adding, when necessary into
Row ultrasonic dissolution assisting；2) triethylamine is added to eggplant-shape bottle, packs tightly bottleneck with sealed membrane or stoppered with bottle stopper, and in magnetic force heating stirring
It is protected from light on device；3) it is lyophilized: eggplant-shape bottle being first put into refrigerator freeze-drying overnight, next day takes out to be placed on freeze dryer and be dehydrated, wait freeze
It is redissolved in pure water after dry, continues freeze-drying overnight, it is repeated multiple times to obtain powder；4.) purify: after activation C18 pillar, (1) is by step
3) powder in is dissolved in NaOH solution, ultrasonic dissolution assisting, extracts every time, squeezes into C18 pillar, after rushed with air it is primary；(2) it takes
NA2CO3Solution, the C18 pillar of step before rinsing；(3) pure water rinsing once before step C18 pillar, then rushed with air it is primary,
Discard filtrate；(4) dehydrated alcohol, the C18 pillar of step, air rush again before rinsing, and collect ethanol liquid in centrifuge tube；(5) will
Penicillin bottle is added in the ethanol rinse liquid being collected into batches, is evaporated with revolving instrument, obtains CB86-DTPAA powder.
(2) trichlorine 1) CB86- DTPAA connection gadolinium after purification: is added into the CB86-DTPAA powder of above-mentioned preparation
Change gadolinium, is dissolved in PBS, is stirred overnight；2) after purified, revolving obtains final product CB86-DTPAA-Gd.Purified is through upper
State the purifying of step (1) purification process.
As optimization, the step 1) of the CB86-DTPAA preparation is: standby 50ml eggplant-shape bottle one, and is encased with masking foil
Body is placed in magnetic heating stirrer；5ml DMSO is added, and is protected from light lower addition 800mg DTPAA, separately weighs 200mg
CB86 is poured slowly into 50ml eggplant-shape bottle by several times, pays attention to slowly instilling, and wants to rotate in drop, it may be necessary to ultrasound
Hydrotropy.
The step 1) of the CB86- DTPAA connection gadolinium after purification is the CB86-DTPAA powder by above-mentioned preparation by 5:
Gadolinium trichloride is added in 1 ratio thereto, is dissolved in the PBS that PH is 7.4-8.0,1200rpm is stirred overnight.
As optimization, the step 2) of CB86-DTPAA preparation is: the triethylamine of 300ul being added to eggplant-shape bottle, uses sealed membrane
It packs tightly bottleneck or is stoppered with bottle stopper, in being protected from light -36h for 24 hours in magnetic heating stirrer.
As optimization, the step 3) of CB86-DTPAA preparation is freeze-drying: eggplant-shape bottle being first put into -80 DEG C of refrigerators and was lyophilized
Night, next day take out and are placed on freeze dryer and are dehydrated, and 20ml pure water is redissolved in after being lyophilized, and continue freeze-drying overnight, repeatedly 3 times powder
End.
As optimization, the activation C18 pillar in the step 4) of CB86-DTPAA preparation is: 10ml absolute alcohol is primary,
10ml water is primary, and air is primary, and moisture is gone out.
As optimization, (1) is the step of the step 4) of CB86-DTPAA preparation: the powder in step 3) is pressed
The ratio of 20mg:10ml is dissolved in the NaOH of 1mmol/l, and ultrasonic dissolution assisting extracts 5-10ml every time, the C18 pillar after squeezing into activation,
It is rushed afterwards with air primary.
As optimization, CB86-DTPAA preparation step 4) the step of (2) be: syringe takes the NA of 1mmol/l2CO3Solution
10ml, the C18 pillar of step before rinsing.
As optimization, (3) are the step of the step 4) of CB86-DTPAA preparation: pure water 10ml rinse it is primary before step
C18 pillar, then rushed once with air, discard filtrate.
As optimization, (4) are the step of the step 4) of CB86-DTPAA preparation: dehydrated alcohol 5ml, step before rinsing
C18 pillar, air rush again, collect ethanol liquid in 50ml centrifuge tube.
As optimization, purification process purifying of the step 2) through above-mentioned steps one in step (2) is purification step referring to step
Suddenly c, d, e in (one) obtains final product CB86-DTPAA-Gd after revolving.Specifically: being extracted in 50ml eggplant-shape bottle with syringe
CB86-DTPAA reacted with GdCl3 after solution 5ml, be injected into the C18 pillar of activation, then use 10ml pure water rinsing
C18 pillar is primary, then is rushed again once with air, discards filtrate, then then rinse C18 pillar with the dehydrated alcohol of 5ml, uses mould
Plain bottle collects dehydrated alcohol leacheate, is placed on revolving instrument and rotates, obtaining powder is final product CB86-DTPAA-Gd.
After adopting the above technical scheme, the present invention targets TSPO magnetic resonance imaging contrast, it can be used for periphery chronic inflammation
Property disease, such as rheumatoid arthritis, the early diagnosis such as rheumatic arthritis and curative effect evaluation have imaging spatial resolution
Height, soft tissue resolution is good, and "dead" advantage.
Detailed description of the invention
Fig. 1 is the synthetic route chart of present invention targeting TSPO magnetic resonance imaging contrast；Fig. 2 is present invention targeting TSPO
The chromatogram that magnetic resonance imaging contrast is identified with HPLC；Fig. 3 be the present invention targeting TSPO magnetic resonance imaging contrast into
The Micro-MRI image results figure of row left side arthritis Balb/c mouse.
Specific embodiment
It is the CB86-DTPAA-Gd made of the following steps that the present invention, which targets TSPO magnetic resonance imaging contrast,.
The preparation of step (1) CB86-DTPAA: 1) to being placed in magnetic heating stirrer and encase body with masking foil
Eggplant-shape bottle DMSO is added, and DTPAA is added, CB86 is separately slowly added to eggplant-shape bottle by several times, and want stirring while adding, it is necessary
Shi Jinhang ultrasonic dissolution assisting；2) triethylamine is added to eggplant-shape bottle, packs tightly bottleneck with sealed membrane or is stoppered with bottle stopper, and heated in magnetic force
It is protected from light on blender；3) it being lyophilized: eggplant-shape bottle being first put into refrigerator freeze-drying overnight, next day takes out to be placed on freeze dryer and be dehydrated,
It is redissolved in pure water after being lyophilized, continues freeze-drying overnight, repeated multiple times obtained powder；4.) it purifies: after activation C18 pillar, (1)
Powder in step 3) is dissolved in NaOH solution, ultrasonic dissolution assisting extracts every time, the C18 pillar after squeezing into activation, after rushed with air
Once；(2) NA is taken2CO3Solution, the C18 pillar of step before rinsing；(3) pure water rinsing once before step C18 pillar, then with empty
Gas punching is primary, discards filtrate；(4) dehydrated alcohol, the C18 pillar of step, air rush again before rinsing, and collect ethanol liquid in centrifugation
Pipe；(5) penicillin bottle is added in the ethanol rinse liquid being collected into batches, is evaporated with revolving instrument, CB86-DTPAA powder is obtained
End.
The CB86- DTPAA connection gadolinium of step (2) after purification: 1) it into the CB86-DTPAA powder of above-mentioned preparation is added
Gadolinium trichloride is dissolved in PBS, is stirred overnight；2) after purified, revolving obtains final product CB86-DTPAA-Gd.It is purified to be
It is purified through above-mentioned steps (one) purification process.High with imaging spatial resolution, soft tissue resolution is good, and "dead" excellent
Point.
The step 1) of the CB86-DTPAA preparation is: standby 50ml eggplant-shape bottle one, and encases body with masking foil, is placed in
In magnetic heating stirrer；5ml DMSO is added, and is protected from light lower addition 800mg DTPAA, it is another to weigh 200mg CB86, delay by several times
Slowly 50ml eggplant-shape bottle is poured into, pays attention to slowly instilling, and want to rotate in drop, it may be necessary to ultrasonic dissolution assisting.CB86-
The step 2) of DTPAA preparation is: the triethylamine of 300ul is added to eggplant-shape bottle, packs tightly bottleneck with sealed membrane or is stoppered with bottle stopper, in
- 36h for 24 hours is protected from light in magnetic heating stirrer.The step 3) of CB86-DTPAA preparation is freeze-drying: be first put into eggplant-shape bottle-
Overnight, next day takes out to be placed on freeze dryer and be dehydrated, and 20ml pure water is redissolved in after being lyophilized, continues to be lyophilized for 80 DEG C of refrigerator freeze-dryings
Night, repeatedly 3 times powder.
The step 1) of the CB86- DTPAA connection gadolinium after purification is the CB86-DTPAA powder by above-mentioned preparation by 5:
Gadolinium trichloride is added in 1 ratio thereto, is dissolved in the PBS that PH is 7.4-8.0,1200rpm is stirred overnight.
Activation C18 pillar in the step 4) of CB86-DTPAA preparation is: 10ml absolute alcohol is primary, and 10ml water is primary,
Air is primary, and moisture is gone out.
The step of step 4) of the CB86-DTPAA preparation (1), is: the ratio by the powder in step 3) by 20mg:10ml
Example is dissolved in the NaOH of 1mmol/l, and ultrasonic dissolution assisting extracts 5-10ml every time, the C18 pillar after squeezing into activation, after with air punching one
It is secondary.The step of step 4) of CB86-DTPAA preparation (2) is: syringe takes the NA of 1mmol/l2CO3Solution 10ml, step before rinsing
Rapid C18 pillar.The step of step 4) of CB86-DTPAA preparation (3) is: the C18 that pure water 10ml rinses primary preceding step is small
Column, then rushed once with air, discard filtrate.The step of step 4) of CB86-DTPAA preparation (4) is: dehydrated alcohol 5ml is rinsed
The C18 pillar of preceding step, air rush again, collect ethanol liquid in 50ml centrifuge tube.
Purification process purifying of the step 2) through above-mentioned steps one in step (2) is purification step referring in step (1)
C, d, e, obtain final product CB86-DTPAA-Gd after revolving.Specifically: extracting CB86- in 50ml eggplant-shape bottle with syringe
DTPAA reacted with GdCl3 after solution 5ml, be injected into the C18 pillar of activation, then use 10ml pure water rinsing C18 pillar
Once, then with air it rushes again once, filtrate is discarded, then then rinse C18 pillar with the dehydrated alcohol of 5ml, with penicillin bottle
Dehydrated alcohol leacheate is collected, is placed on revolving instrument and rotates, obtaining powder is final product CB86-DTPAA-Gd.
High with imaging spatial resolution, soft tissue resolution is good, and "dead" advantage.
The step of preparation method of targeting TSPO magnetic resonance imaging contrast of the present invention, is as follows.
(1) preparation of CB86-DTPAA: 1) to the eggplant for being placed in magnetic heating stirrer and being encased with masking foil body
DMSO is added in shape bottle, and DTPAA is added, and CB86 is separately slowly added to eggplant-shape bottle by several times, and want stirring while adding, when necessary into
Row ultrasonic dissolution assisting；2) triethylamine is added to eggplant-shape bottle, packs tightly bottleneck with sealed membrane or stoppered with bottle stopper, and in magnetic force heating stirring
It is protected from light on device；3) it is lyophilized: eggplant-shape bottle being first put into refrigerator freeze-drying overnight, next day takes out to be placed on freeze dryer and be dehydrated, wait freeze
It is redissolved in pure water after dry, continues freeze-drying overnight, it is repeated multiple times to obtain powder；4.) purify: after activation C18 pillar, (1) is by step
3) powder in is dissolved in NaOH solution, ultrasonic dissolution assisting, extracts every time, squeezes into C18 pillar, after rushed with air it is primary；(2) it takes
NA2CO3Solution, the C18 pillar of step before rinsing；(3) pure water rinsing once before step C18 pillar, then rushed with air it is primary,
Discard filtrate；(4) dehydrated alcohol, the C18 pillar of step, air rush again before rinsing, and collect ethanol liquid in centrifuge tube；(5) will
Penicillin bottle is added in the ethanol rinse liquid being collected into batches, is evaporated with revolving instrument, obtains CB86-DTPAA powder.
(2) trichlorine 1) CB86- DTPAA connection gadolinium after purification: is added into the CB86-DTPAA powder of above-mentioned preparation
Change gadolinium, is dissolved in PBS, is stirred overnight；2) after purified, revolving obtains final product CB86-DTPAA-Gd.Purified is through upper
State the purifying of step (1) purification process.
The step 1) of the CB86-DTPAA preparation is: standby 50ml eggplant-shape bottle one, and encases body with masking foil, is placed in
In magnetic heating stirrer；5ml DMSO is added, and is protected from light lower addition 800mg DTPAA, it is another to weigh 200mg CB86, delay by several times
Slowly 50ml eggplant-shape bottle is poured into, pays attention to slowly instilling, and want to rotate in drop, it may be necessary to ultrasonic dissolution assisting.CB86-
The step 2) of DTPAA preparation is: the triethylamine of 300ul is added to eggplant-shape bottle, packs tightly bottleneck with sealed membrane or is stoppered with bottle stopper, in
- 36h for 24 hours is protected from light in magnetic heating stirrer.The step 3) of CB86-DTPAA preparation is freeze-drying: be first put into eggplant-shape bottle-
Overnight, next day takes out to be placed on freeze dryer and be dehydrated, and 20ml pure water is redissolved in after being lyophilized, continues to be lyophilized for 80 DEG C of refrigerator freeze-dryings
Night, repeatedly 3 times powder.Activation C18 pillar in the step 4) of CB86-DTPAA preparation is: 10ml absolute alcohol is primary,
10ml water is primary, and air is primary, and moisture is gone out.
The step 1) of the CB86- DTPAA connection gadolinium after purification is the CB86-DTPAA powder by above-mentioned preparation by 5:
Gadolinium trichloride is added in 1 ratio thereto, is dissolved in the PBS that PH is 7.4-8.0,1200rpm is stirred overnight.
The step of step 4) of the CB86-DTPAA preparation (1), is: the ratio by the powder in step 3) by 20mg:10ml
Example is dissolved in the NaOH of 1mmol/l, and ultrasonic dissolution assisting extracts 5-10ml every time, the C18 pillar after squeezing into activation, after with air punching one
It is secondary.The step of step 4) of CB86-DTPAA preparation (2) is: syringe takes the NA of 1mmol/l2CO3Solution 10ml, step before rinsing
Rapid C18 pillar.The step of step 4) of CB86-DTPAA preparation (3) is: the C18 that pure water 10ml rinses primary preceding step is small
Column, then rushed once with air, discard filtrate.The step of step 4) of CB86-DTPAA preparation (4) is: dehydrated alcohol 5ml is rinsed
The C18 pillar of preceding step, air rush again, collect ethanol liquid in 50ml centrifuge tube.
Purification process purifying of the step 2) through above-mentioned steps one in step (2) is purification step referring in step (1)
C, d, e, obtain final product CB86-DTPAA-Gd after revolving.Specifically: extracting CB86- in 50ml eggplant-shape bottle with syringe
DTPAA reacted with GdCl3 after solution 5ml, be injected into the C18 pillar of activation, then use 10ml pure water rinsing C18 pillar
Once, then with air it rushes again once, filtrate is discarded, then then rinse C18 pillar with the dehydrated alcohol of 5ml, with penicillin bottle
Dehydrated alcohol leacheate is collected, is placed on revolving instrument and rotates, obtaining powder is final product CB86-DTPAA-Gd.
The present invention targets TSPO magnetic resonance imaging contrast and preparation method more specifically.
(1) preparation of CB86-DTPAA, 1. for 50ml eggplant-shape bottle one, and encases body with masking foil, is placed in magnetic force
On heating stirrer.5ml DMSO is added, and 800mg DTPAA(is added and is preferably protected from light), it is another to weigh 200mg CB86, delay by several times
Slowly 50ml eggplant-shape bottle is poured into, pays attention to slowly instilling, and want to rotate in drop.It when necessary can be with ultrasonic dissolution assisting.
2. the triethylamine of 300ul is added to eggplant-shape bottle, pack tightly bottleneck (or being stoppered with bottle stopper) with sealed membrane, in magnetic force plus
- 36h for 24 hours is protected from light on thermal agitation device.3. eggplant-shape bottle is first put into -80 DEG C of refrigerator freeze-dryings overnight by freeze-drying, next day taking-up is set
In being dehydrated on freeze dryer, it is redissolved in 20ml pure water after being lyophilized, continues freeze-drying overnight, 3 times repeatedly.4. purifying, first activates
The NaOH solution of C18 pillar (10ml absolute alcohol is primary, and 10ml water is primary, and air is primary, and moisture is gone out) preparation 1mmol/l
With the NA2CO3 solution of 1mmol/l.A., powder in step 3 is dissolved in the NaOH of 1mmol/l in the ratio of 20mg:10ml, is surpassed
Sound hydrotropy extracts 5 ~ 10ml every time, squeezes into C18 pillar, after rushed with air it is primary.B. syringe takes the NA2CO3 of 1mmol/l molten
The C18 pillar of liquid 10ml, rinsing step a.C. pure water 10ml rinses the C18 pillar of a step b, then primary, abandoning is rushed with air
Go filtrate.D. the C18 pillar of dehydrated alcohol 5ml, rinsing step C, air rush again, collect ethanol liquid in 50ml centrifuge tube.e.
Penicillin bottle is added in the ethanol rinse liquid being collected into batches, is evaporated with revolving instrument, obtains CB86-DTPAA powder.
(2) CB86- DTPAA connection gadolinium after purification: 1. are added the powder of above-mentioned preparation in the ratio of 5:1 thereto
Enter gadolinium trichloride (GdCl3), it is dissolved in the PBS that PH is 7.4 ~ 8.0,1200rpm is stirred overnight.2. after purified, purifying
Step obtains final product CB86-DTPAA-Gd after revolving referring to c, d, e in step (1).Step 2) warp in step (2)
The purification process purifying of above-mentioned steps one is c, d, e in purification step reference step (1), obtains final product CB86- after revolving
DTPAA-Gd.Specifically: the solution 5ml after being reacted with CB86-DTPAA in syringe extraction 50ml eggplant-shape bottle with GdCl3, injection
It is then primary with the pure water rinsing C18 pillar of 10ml into the C18 pillar of activation, then rushed again once with air, filtrate is discarded,
C18 pillar is then rinsed with the dehydrated alcohol of 5ml again, collects dehydrated alcohol leacheate with penicillin bottle, is placed on revolving instrument
Revolving, obtaining powder is final product CB86-DTPAA-Gd.CB86-DTPAA-Gd synthetic line is as shown in Figure 1.
Gradient condition used (flow velocity is 0.5 ml/min) is such as Fig. 2 institute to be identified to product CB86-DTPAA-Gd with HPLC
Show: the appearance time of the product is about are as follows: 25.7min.
Arthritis animal model MRI imaging: Micro- of the CB86-DTPAA-Gd in left side arthritis Balb/c mouse
MRI image results as shown in figure 3, from figure 3, it can be seen that as a control group with magnevist (Magnevist Solution injection),
For the mouse arthritis disease model constructed with modeling mode of the same race, pass through tail vein injection same concentrations, same dose
CB86-DTPAA-Gd and magnevist compare their signal to noise ratio (snr), it can be seen that on t1 weighted image, CB86-
DTPAA-Gd has the function of stronger signal enhancing at left side arthritis position.And by inhibiting to test visible CB86-
DTPAA-Gd has stronger selectively targeted ability to arthritis position.This for arthritis early detection, by stages
Evaluation, curative effect monitoring, prognostic evaluation have huge potential.A:CB86-DTPAA-Gd in Fig. 3, SNR value 28；B: Ma Genwei
It is aobvious, SNR value 19.5；C: inhibit test, SNR value 10.8.
In short, after adopting the above technical scheme, the present invention targets TSPO magnetic resonance imaging contrast and preparation method is available
In periphery chronic inflammation disease, such as rheumatoid arthritis, the early diagnosis such as rheumatic arthritis and curative effect evaluation have aobvious
Image space high resolution, soft tissue resolution is good, and "dead" advantage.
Claims (10)
1. a kind of targeting TSPO magnetic resonance imaging contrast, it is characterised in that the CB86-DTPAA-Gd made of the following steps:
The preparation of step (1) CB86-DTPAA: 1) to the eggplant for being placed in magnetic heating stirrer and being encased with masking foil body
DMSO is added in shape bottle, and DTPAA is added, and CB86 is separately slowly added to eggplant-shape bottle by several times, and want stirring while adding, when necessary into
Row ultrasonic dissolution assisting；2) triethylamine is added in step eggplant-shape bottle upwards, packs tightly bottleneck with sealed membrane or is stoppered with bottle stopper, and heats in magnetic force
It is protected from light on blender；3) it is lyophilized: upper step eggplant-shape bottle being first put into refrigerator freeze-drying overnight, next day, which takes out to be placed on freeze dryer, to be taken off
Water is redissolved in pure water after being lyophilized, and continues freeze-drying overnight, repeated multiple times to obtain powder；4.) it purifies: after activation C18 pillar, (1)
Powder in step 3) is dissolved in NaOH solution, ultrasonic dissolution assisting extracts every time, the C18 pillar after squeezing into activation, after rushed with air
Once；(2) NA is taken2CO3Solution, the C18 pillar of step before rinsing；(3) pure water rinsing once before step C18 pillar, then with empty
Gas punching is primary, discards filtrate；(4) dehydrated alcohol, the C18 pillar of step, air rush again before rinsing, and collect ethanol liquid in centrifugation
Pipe；(5) penicillin bottle is added in the ethanol rinse liquid being collected into batches, is evaporated with revolving instrument, CB86-DTPAA powder is obtained
End；
The CB86- DTPAA connection gadolinium of step (2) after purification: 1) into the CB86-DTPAA powder of above-mentioned preparation it is added trichlorine
Change gadolinium, is dissolved in PBS, is stirred overnight；2) after purified, revolving obtains final product CB86-DTPAA-Gd.
2. targeting TSPO magnetic resonance imaging contrast according to claim 1, it is characterised in that the CB86-DTPAA preparation
Step 1) be: standby 50ml eggplant-shape bottle one, and encase body with masking foil is placed in magnetic heating stirrer；5ml is added
DMSO, and it is protected from light lower addition 800mg DTPAA, it is another to weigh 200mg CB86, it is poured slowly into 50ml eggplant-shape bottle by several times, pays attention to delay
It slowly instills, and wants to rotate in drop, it may be necessary to ultrasonic dissolution assisting；
The step 1) of the CB86- DTPAA connection gadolinium after purification is the CB86-DTPAA powder by above-mentioned preparation by 5:1's
Gadolinium trichloride is added in ratio thereto, is dissolved in the PBS that PH is 7.4-8.0,1200rpm is stirred overnight.
3. targeting TSPO magnetic resonance imaging contrast according to claim 2, it is characterised in that the step of CB86-DTPAA preparation
It is rapid 2) to be: the triethylamine of 300ul being added to eggplant-shape bottle, packs tightly bottleneck with sealed membrane or is stoppered with bottle stopper, in magnetic force heating stirring
- 36h for 24 hours is protected from light on device.
4. targeting TSPO magnetic resonance imaging contrast according to claim 3, it is characterised in that the step of CB86-DTPAA preparation
3) rapid is freeze-drying: eggplant-shape bottle being first put into -80 DEG C of refrigerator freeze-dryings overnight, next day takes out to be placed on freeze dryer and be dehydrated, after being lyophilized
Be redissolved in 20ml pure water, continue freeze-drying overnight, repeatedly 3 times powder.
5. targeting TSPO magnetic resonance imaging contrast according to claim 4, it is characterised in that the step of CB86-DTPAA preparation
It is rapid 4) in activation C18 pillar be: 10ml absolute alcohol is primary, and 10ml water is primary, and air is primary, and moisture is gone out.
6. -5 any targeting TSPO magnetic resonance imaging contrast according to claim 1, it is characterised in that the CB86-
The step of step 4) of DTPAA preparation (1), is: the powder in step 3) is dissolved in 1mmol/l's in the ratio of 20mg:10ml
NaOH, ultrasonic dissolution assisting extract 5-10ml every time, the C18 pillar after squeezing into activation, after rushed with air it is primary.
7. targeting TSPO magnetic resonance imaging contrast according to claim 6, it is characterised in that the step of CB86-DTPAA preparation
It is rapid 4) the step of (2) be: syringe takes the NA of 1mmol/l2CO3Solution 10ml, the C18 pillar of step before rinsing.
8. targeting TSPO magnetic resonance imaging contrast according to claim 7, it is characterised in that the step of CB86-DTPAA preparation
It is rapid 4) the step of (3) be: pure water 10ml rinse it is primary before step C18 pillar, then rushed with air primary, discard filtrate.
9. targeting TSPO magnetic resonance imaging contrast according to claim 8, it is characterised in that the step of CB86-DTPAA preparation
It is rapid 4) the step of (4) be: dehydrated alcohol 5ml, the C18 pillar of step before rinsing, air rushes again, collect ethanol liquid in 50ml from
Heart pipe.
10. targeting the preparation method of TSPO magnetic resonance imaging contrast described in claim 1, it is characterised in that steps are as follows:
(1) preparation of CB86-DTPAA: 1) to the eggplant-shape bottle for being placed in magnetic heating stirrer and being encased with masking foil body
DMSO is added, and DTPAA is added, CB86 is separately slowly added to eggplant-shape bottle by several times, and want stirring while adding, is surpassed when necessary
Sound hydrotropy；2) triethylamine is added to eggplant-shape bottle, packs tightly bottleneck with sealed membrane or stoppered with bottle stopper, and in magnetic heating stirrer
It is protected from light；3) it is lyophilized: eggplant-shape bottle being first put into refrigerator freeze-drying overnight, next day takes out to be placed on freeze dryer and be dehydrated, after being lyophilized
It is redissolved in pure water, continues freeze-drying overnight, it is repeated multiple times to obtain powder；4.) purify: after activation C18 pillar, (1) will be in step 3)
Powder be dissolved in NaOH solution, ultrasonic dissolution assisting extracts every time, squeezes into C18 pillar, after rushed with air it is primary；(2) NA is taken2CO3It is molten
Liquid, the C18 pillar of step before rinsing；(3) pure water rinsing once before step C18 pillar, then rushed with air primary, discard filter
Liquid；(4) dehydrated alcohol, the C18 pillar of step, air rush again before rinsing, and collect ethanol liquid in centrifuge tube；(5) it will be collected into
Ethanol rinse liquid penicillin bottle is added in batches, be evaporated with revolving instrument, obtain CB86-DTPAA powder；
(2) CB86- DTPAA connection gadolinium after purification: 1) being added gadolinium trichloride into the CB86-DTPAA powder of above-mentioned preparation,
It is dissolved in PBS, is stirred overnight；2) after purified, revolving obtains final product CB86-DTPAA-Gd.
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