CN105001274B - Complex, the Preparation method and use of a kind of Glucosamine derived ligand compound and preparation method, three carbonyl Tc 99ms mark - Google Patents
Complex, the Preparation method and use of a kind of Glucosamine derived ligand compound and preparation method, three carbonyl Tc 99ms mark Download PDFInfo
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Abstract
The invention provides a kind of Glucosamine derived ligand compound and preparation method thereof.Present invention also offers a kind of complex of three carbonyls Tc 99m mark, and it has the structure as shown in formula (B), and present invention also offers the preparation method and purposes of three carbonyl Tc 99ms mark complex.The Glucosamine derived ligand compound of the present invention is coupled Glucosamine with 2,3 diaminopropionic acids, and raw material is easy to get, preparation method is easy.The three carbonyl Tc 99ms mark complex of the present invention is formed by the Glucosamine derived ligand compound and the carbonyl Tc 99m of radioactivity three, chemical stability is strong, preparation is simple, chemical purity is high, biological property is good, can be used for preparing new tumour SPECT developers.
Description
Technical field
The present invention relates to clinical nuclear medicine technical field, and in particular to a kind of Glucosamine derived ligand compound and
Its preparation method and as the technetium-99 m labeled complex of three carbonyls obtained by the Glucosamine derived ligand compound,
Preparation method and its usage.
Background technology
Glucose is important substance needed for energetic supersession, into after blood circulation, relies on glucose transporter in vivo
(glucose transporter, GLUT) enters cell.The abnormal energy metabolism of tumour cell, compared with normal cell, its
Glucose uptake is higher.Therefore intake of the tumor tissues to glucose and the like can be utilized, prepares radioactive grape
Sugared glycometabolism molecular probe, noninvasive test and evaluation for in-vivo tumour.18F- deoxyglucoses (18F-FDG it is) by grape
Hydroxyl on sugared 2nd carbon atom by18F substitutes, and has no effect on GLUTs to its transporting mechanism, thus after entering in vivo by positioned at
The GLUTs mediations of cell membrane surface enter cell, and Phosphorylated products 6- phosphoric acid FDG is had differences due to structure, it is impossible to by phosphorus
It is 6- phosphoric acid-fluoro fructose that sour hexose isomerase, which continues catalyzed transitions, so as to can not further be metabolized, so as in tumour cell
Middle accumulation.18F-FDG is PET developers most widely used at present, is used clinically for the nuclear medicine image of glycometabolism degree.Its
Recurrence and bad after the application in terms of tumour mainly includes malignant and benign lesion diagnosis, clinical stage, discriminating oncotherapy
Extremely, malignancy, prognosis and therapeutic evaluation etc. are evaluated.
Technetium -99m (99mTc) it is a kind of SPECT nucleic with excellent nucleic property.In recent years99mThe glucose of Tc marks
Metabolic molecule probe is increasingly becoming the focus of research.It is difunctional by being introduced in the diverse location of glucose or Glucosamine
Articulation agent, and pass through99mTc is marked, and has obtained a variety of different types of labels.Including：99mTc-ECDG、99mTc-
DTPA-DG、99mTcN-DGDTC、 99mTcO-DGDTC etc..Glucosamine molecules spread out because its amino part can effectively carry out coordination
It is raw, receive substantial amounts of research concern in the research of glucose tracers, many documents are had proven to after nitrogen end functionalization still
It can be identified and over-expressed by GLUTs, and the functional group size modified influences less.Three carbonyl technetium -99m cores
(fac-[99mTc(CO)3]+) there is advantage such as small size and molecular weight, good water solubility and stability.Research report at present
The aminoglucose sugar derivatives of three carbonyl technetium -99m cores mark, mainly introduces tridentate ligand on 2 bit aminos.Such as N-
(2'- hydroxybenzyls) -2- amino deoxies glucose (N- (2'-Hydroxybenzyl) -2-amino-2-deoxy-D-
Glucose), it is coordinated using phenol oxygen, ammonia nitrogen and 3 sugared hydroxyl oxygens and carbonyl technetium, gained label is excessive half
Stability is performed poor in cystine and histidine competitive assay；Lutidines amine is introduced in Glucosamine
(dipicolylamine) derivative is marked, gained mark as bifunctional linking reagent by three carbonyl technetium -99m cores
Thing shows higher stability in cysteine and histidine competitive assay in vitro；Ring penta 2 is introduced in Glucosamine
Alkene (cyclopentadienyl) derivative can also cross three carbonyl technetium -99m cores and be marked as bifunctional linking reagent, its
Three rhenium carbonyls (Re) complex is capable of the phosphorylation of Competitive assays glucose, Ki in the experiment of hexokinase Competitive assays
Value is smaller than FDG three times, it is shown that it has certain bioactivity.
18F-FDG has a wide range of applications in nuclear medicine, still18F half-life period is relatively short, is needed in preparation process
The reasons such as cyclotron is relatively expensive use upper still limited.Compared with PET, SPECT equipment is more popular and cheap.Hair
Exhibition SPECT nucleic is particularly technetium-99 m labeled glucosan derivative for extension Nuclear medicine means, increase clinical practice
Have great importance.Find at present a kind of technetium-99 m labeled with higher label stability and good biological performance
Aminoglucose sugar derivatives, the SPECT for tumour are imaged, and are the important topics that the art needs to solve.
The content of the invention
To overcome drawbacks described above existing for existing tumour SPECT developers, there is provided a kind of chemical stability is good, biological
Can excellent new SPECT developers, an object of the present invention is to provide a kind of SPECT developers that can be used conveniently to prepare
Glucosamine derived ligand compound and preparation method thereof.
It is a further object of the present invention to provide a kind of technetium-99 m labeled complex of three carbonyl, and preparation method thereof and use
In the purposes for preparing SPECT developers.
Glucosamine derived ligand compound provided by the invention, it has the structure as shown in formula (A),
Wherein ,-(CH2)n- it is straight chain or the structure comprising side chain, n represents 2~8 integer.
In Glucosamine derived ligand compound provided by the invention, preferably described-(CH2)n- it is linear chain structure, n tables
Show 2~5 integer.
The preparation method of the Glucosamine derived ligand compound provided by the invention, comprises the following steps：
(1) it is that initiation material is reduced to the tertiary fourth of 2- amino -2- cyanoacetic acids with 2- oximido -2- t-butylcyanoacetates (I)
Ester (II)；
(2) amino of the 2- amino -2- t-butylcyanoacetates (II) obtained by step (1) is carried out into radical protection to obtain
Midbody compound (III), and with halogenated carboxylic ester R ' OOC (CH2)nX reacts to obtain midbody compound (IV-n)；
(3) cyano reduction in the midbody compound (IV-n) obtained by step (2) is amino and carries out radical protection
Obtain midbody compound (V-n)；
(4) by the hydrolysis of ester group of the midbody compound (V-n) obtained by step (3) obtain midbody compound (VI-
n)；
(5) midbody compound (VI-n) obtained by step (4) and four acetoxyl group Glucosamines are reacted in obtaining
Intermediate compounds therefor (VII-n)；
(6) midbody compound (VII-n) obtained by step (5) is subjected to group deprotection, hydrolysis and obtains the ammonia
Base Derived from D-Glucose ligand compound；
In above-mentioned preparation process, the α amino at 2,3- diaminopropionic acids (DAP) end is protected using Boc, and carboxyl uses tertiary fourth
Ester is protected, and such blocking group can be compared with sloughing under temperate condition, and does not influence the acetyl protecting group on glucose
(A)。
Synthetic route is as follows：
In above-mentioned preparation method, the halogenated carboxylic ester R ' OOC (CH2)nIn X, X expressions F, Cl or Br, R ' expressions C1~
The group such as the straight or branched alkyl of C10 straight or branched alkyl, preferably C1~C5, more preferably methyl, ethyl, isopropyl.
The technetium-99 m labeled complex of three carbonyl provided by the invention, it has the structure as shown in formula (B),
Wherein ,-(CH2)n- it is straight chain or the structure comprising side chain, n represents 2~8 integer, and M is represented99mTc。
In the technetium-99 m labeled complex of three carbonyl provided by the invention ,-(CH2)n- it is linear chain structure, n represents 2~5
Integer.
The preparation method of the technetium-99 m labeled complex of three carbonyl provided by the invention, comprises the following steps：
(1) prepare three carbonyl technetium -99m intermediate hydrated ions fac- [99mTc(CO)3(H2O)3]+；
(2) by the fac- of gained [99mTc(CO)3(H2O)3]+With the Glucosamine derived ligand of claim 1 or 2
The complex is made in compound reaction.
In above-mentioned preparation method, in the step (2), fac- [99mTc(CO)3(H2O)3]+Described in claim 1 or 2
Glucosamine derived ligand compound is heated to 70~100 DEG C of 5~30min of reaction and the cooperation is made in neutral conditions
Thing.
In above-mentioned preparation method, the neutrallty condition is the phosphate buffer that pH value is 7.
In above-mentioned preparation method, intermediate hydrated ion fac- [99mTc(CO)3(H2O)3]+Preparation can use it is existing
Preparation method.
Preferably can be procedure below in above-mentioned preparation method：
A, sodium borohydride, sodium potassium tartrate tetrahydrate, sodium carbonate are placed in penicillin bottle, aluminium lid seals after covering plug, uses
It is evacuated pin and extracts air, is passed through CO gases 10-20min thereto.
B, will be from medical99Mo-99mEluted in Tc generators acquisition Sodium Pertechnetate Na [99mTcO4] leacheate, add step
In reaction bulb prepared by rapid a, 10~60min is reacted under 70~100 DEG C of heating conditions after shaking up dissolving, that is, obtains three carbonyls
Technetium -99m intermediates fac- [99mTc(CO)3(H2O)3]+。
C, hydrochloric acid will be added in step b resulting solutions, regulation pH value is 6~8.
D, 2,3- diaminopropionic acids are coupled into aminoglucose carbohydrate ligands (DAP-n-DG) to be dissolved in cushioning liquid, add step
In rapid c solution, 5~30min is reacted under 70~100 DEG C of heating conditions.
The concentration of hydrochloric acid used in above-mentioned steps c is 1mol/L.
Buffer solution described in above-mentioned steps d can be 0.5mol/L pH7.0 phosphate buffers.
It is used to prepare tumour SPECT developers present invention also offers the technetium-99 m labeled complex of three carbonyl
Purposes.
The Glucosamine derived ligand compound of the present invention is coupled Glucosamine, raw material with 2,3- diaminopropionic acids
It is easy to get, prepares simplicity.The methylene with certain length is also included in the Glucosamine derived ligand compound structure of the present invention
Base bridge linkage group, it is possible to reduce due to chelation group mark radioactive metal after volume it is larger and for glucose performance produce
Influence so that mark coordinates physical performance more stable.
The technetium-99 m labeled complex of three carbonyls of the present invention is by the Glucosamine derived ligand compound and radioactivity
Three carbonyl technetium -99m are formed, and are had the characteristics that small volume, are facilitated penetration of cell membrane, and chemical stability is strong, it is simple to prepare,
Chemical purity is high, biological property is good, can be used for preparing new tumour SPECT developers.
Brief description of the drawings
Fig. 1 is the synthetic route chart of the Glucosamine derived ligand compound described in the embodiment of the present invention.
Embodiment
Below by embodiment, the present invention is described in detail, so that the features and advantages of the present invention become apparent from.But should
This points out that embodiment is used for the design for understanding the present invention, and the scope of the present invention is not limited only to reality listed herein
Apply example.
Such as be not particularly illustrated, raw material used in embodiment be commercially available prod, it is used operation be this area
Routine operation.
The preparation of the Glucosamine derived ligand compound (DAP-n-DG) of embodiment 1
As shown in figure 1, comprise the following steps：
(1) synthesis of 2- amino -2- t-butylcyanoacetates (II)
11.98g raw material 2- oximido -2- t-butylcyanoacetates (I) are added in 88mL deionized waters, added pre-
The 80mL saturated sodium bicarbonate aqueous solutions first prepared, stir lower reaction and carry out about 40min.Reaction bulb is placed in 40 DEG C of oil bath pans
In, it is added portionwise into the common 33g of sodium hydrosulfite.React about 9h.15g sodium chloride is added, fully dissolving is cooled to room temperature.Mixed liquor is with two
Chloromethanes is extracted repeatedly (every time with 50mL), and organic extractant phase liquid is dried with anhydrous magnesium sulfate, and revolving removes solvent
Obtain brown oil (II) 2.10g, yield 19%.
(2) synthesis of 2.2- (N- t-butoxycarbonyl aminos) -2- t-butylcyanoacetates (III)
8.94mmol 2- amino -2- t-butylcyanoacetates (II) are added in 12mL toluene, after abundant dissolving
The DIPEA (DIPEA) that volume is 1.477mL is added, it is then disposable to add di-tert-butyl dicarbonate
(Boc2O) 3.44g, reaction vessel is placed on heated overnight at reflux in 115 DEG C of oil bath pans.Add 5mL water and extract reaction of going out, use
50mL × 3 time ethyl acetate extraction.Using column chromatography, pure pale yellow oil (III) 1.483g (5.8mmol) is obtained,
Yield is 65%.
1H NMR(400MHz,CDCl3)δ:1.57(s,18H,tBu),5.35(m,1H,H-C)。
(3) synthesis of midbody compound (IV-n)
50mL Schlenk bottles are removed water into deoxygenation under vacuum line, the hydrogen sodium that content is 60% is added under argon gas protection
74.1mg(2.75mmol).By 2- (N- t-butoxycarbonyl aminos) -2- t-butylcyanoacetates (III) 281.9mg
(1.1mmol) with adding in reaction bulb after steaming tetrahydrofuran 20mL dissolving again, back flow reaction half is small under the conditions of 60 DEG C of oil bath
When.
When (3-1) n is 2, the synthesis of midbody compound (IV-2)
Ethyl bromide 298mg (1.5eq.) is added, reacts the heated overnight at reflux under the conditions of 70 DEG C.Solvent is spin-dried for, is added
Extracted after water with 50mL × 3 time ethyl acetate, with saturated common salt water washing, magnesium sulfate spins off solvent after drying and obtains brown color
Oily crude product, is purified by column chromatography, and product is yellow oil (IV-2) 169mg (0.5mmol), and reaction yield is
45%.
1H NMR(400MHz,CDCl3)δ:1.27(s,3H,CH3), 1.44~1.54 (2s, 18H, tBu), 1.98~
2.04(m,2H,CH2), 2.35~2.40 (m, 2H, CH2),4.13(m,2H, CH2-O)。
When (3-2) n is 3, the synthesis of midbody compound (IV-3)
Bromobutyrate 321.8mg (1.5eq.) is added, reacts the heated overnight at reflux under the conditions of 70 DEG C.It is spin-dried for solvent,
Add and extracted with 50mL × 3 time ethyl acetate after water, with saturated common salt water washing, magnesium sulfate spun off after drying solvent obtain it is pale brown
Color oily crude product 378.9mg.Column chromatography purifying is carried out to crude product, product is yellow oil (IV-3) 202.9mg
(0.55mmol), reaction yield 50%.
1H NMR(400MHz,CDCl3)δ:1.26(s,3H,CH3),1.46(s,9H,tBu), 1.53(s,9H,tBu),
1.79(m,2H,CH2), 2.00~2.20 (m, 2H, CH2),2.38(m,2H, CH2),4.13(m,2H,CH2-O)。
When (3-3) n is 5, the synthesis of midbody compound (IV-5)
Bromocaproic acid ethyl ester 368mg (1.5eq.) is added, reacts the heated overnight at reflux under the conditions of 70 DEG C.Solvent is spin-dried for, is added
Extracted after water with 50mL × 3 time ethyl acetate, with saturated common salt water washing, magnesium sulfate spins off solvent after drying and obtains brown color
Oily crude product 378.9mg.Column chromatography purifying is carried out to crude product, product is yellow oil (IV-5) 182mg
(0.46mmol), reaction yield 42%.
1H NMR(400MHz,CDCl3)δ:1.27(s,3H,CH3), 1.44~1.54 (2s, 18H, tBu), 1.78~
2.03(m,6H,CH2), 2.35~2.40 (m, 2H, CH2),4.13(m,2H, CH2-O)。
(4) synthesis of midbody compound (V-n)
15mL methanol is added after 50mL Schlenk bottles are removed water into deoxygenation applying argon gas under vacuum line.1.38mmol is former
Material (IV-n) is added in reaction bulb, weighs NiCl249.2mg(0.21mmol), Boc2O 602.37mg (2.76mmol) are respectively
It is added in reaction bulb, ice bath adds sodium borohydride 426.62mg (11.27mmol) reactions overnight after abundant dissolving.Add
Diethyl triamine 0.264mL, reaction dissolvent is spin-dried for after reacting 1h.50mL × 3 time second is used after adding saturated sodium bicarbonate aqueous solution
Acetoacetic ester is extracted, and extract is washed twice with saturated common salt, and magnesium sulfate rotates to obtain oily crude product after drying.With post layer
Analysis purifies to product, and product is yellow oil (V-n), and reaction yield is about 56%.
1H NMR(400MHz,CDCl3)(V-3)δ:1.24(m,3H,CH3), 1.43~1.60 (m, 27H, tBu), 1.61
~1.75 (m, 6H, CH2), 2.25~2.30 (m, 2H, CH2),3.66(m, 2H,CH2-N),4.11(m,2H,CH2-O)。ESI-
MS:m/z 497.2722[M+Na]。
(5) synthesis of midbody compound (VI-n)
After reaction raw materials (V-n) (0.126mmol) are dissolved with 10 μ L methanol, it is that 1 M sodium hydroxides are water-soluble to add concentration
The μ L of liquid 315, nearly 8h is stirred in oil bath pan under 60 DEG C of heating conditions.Place room temperature cooling.Adjusted under ice bath with 1M aqueous hydrochloric acid solutions
To slant acidity (pH value is in 4-6), 10mL water is added, product is repeatedly extracted with dichloromethane, extract is washed with saturated common salt
Once, magnesium sulfate is dried, and white solid product (VI-n) is obtained after being spin-dried for, reaction yield is about 78%.
1H NMR(400MHz,CDCl3)(VI-3)δ:1.40~1.46 (m, 27H, tBu), 1.58,1.72 (m, 2H,
CH2),2.16(m,2H,CH2), 2.30~2.38 (m, 2H, CH2),3.67(m,2H, CH2-N)。ESI-MS:m/z 469.2442
[M+Na]。
(6) synthesis of midbody compound (VII-n)
Four acetoxyl group glucose hydrochloride salt 0.144g (0.37mmol) are added into 6mL dichloromethane, add 4- diformazan ammonia
Yl pyridines (DMAP) 0.135g (1.10mmol).Stir add after half an hour raw material (VI-n) 0.44mmol and 1- ethyls-
(3- dimethylaminopropyls) phosphinylidyne diimmonium salt hydrochlorate (EDC) 0.085g (0.44mmol), sealing reaction carry out 12h.Add
10mL dichloromethane, organic phase are cleaned with 5mL × 2 time saturated sodium bicarbonate aqueous solution, and magnesium sulfate rotates after drying, column chromatography
Method purifies to product, obtains white solid powder (VII-n), and reaction yield is about 47%.
1H NMR(400MHz,CDCl3)(VII-3)δ:1.39~1.45 (m, 27H, tBu), 1.62 (m, 2H, CH2),
2.01 (s, 8H, Ac), 2.07~2.09 (m, 4H, CH2),3.62(m,2H, CH2-N),3.79,4.10,4.26,4.82,5.59,
5.91(m,7H,glucose)。ESI-MS:m/z 798.3546[M+Na]。
(7) synthesis of midbody compound (VIII-n)
0.05mmol raw materials (VII-n) are dissolved in 0.333mL dichloromethane, 0.333mL is slowly added dropwise thereto
TFA, 5h is reacted at room temperature.Revolving removes reaction dissolvent.The white solid for being spin-dried for obtaining is dissolved in a small amount of dichloromethane, added
Enter into 50mL ether to obtain white flock insoluble matter.Dissolved after filtering with a small amount of dichloromethane, add TEA and neutralize instead
Half an hour is answered, white solid powder (VIII-n) is obtained after being spin-dried for, reaction yield is about 36%.
1H NMR(400MHz,D2O)(VIII-3)δ:1.64(m,2H,CH2),2.03(s,8H, Ac),2.08(m,2H,
CH2),2.23(m,2H,CH2),3.50(m,2H,CH2- N), 3.80,4.15,4.30,5.05,5.36~5.51 (m, 7H,
glucose)。
(8) DAP-n-DG synthesis
Raw material (VIII-n) 20mg is taken, it is 2molL that 0.5mL concentration is added in penicillin bottle-1Hydrochloric acid, gland.
10min is reacted under 95 DEG C of metal baths, sloughs the Ac protections functional group on glucose.It is 2molL to add 0.5mL concentration-1's
NaOH solution is neutralized to pH 7 or so.Obtain DAP-n-DG.
The preparation of the technetium-99 m labeled complex of the carbonyl of embodiment 2 three
Sodium borohydride 5.5mg, sodium potassium tartrate tetrahydrate 20mg, sodium carbonate 4mg are weighed respectively to be placed in penicillin bottle, are covered
Aluminium lid seals after plug, extracts air with pumping pin, is passed through CO gases 10-20min thereto.Will be from medical99Mo-99mTc is sent out
Eluted in raw device acquisition Sodium Pertechnetate Na [99mTcO4] leacheate 2mL (about 2mCimL-1), add in reaction bulb, shake up molten
30min is reacted after solution under 75 DEG C of metal bath heating conditions, that is, obtains three carbonyl technetium -99m intermediates.Add 1molL-1Salt
Acid, regulation pH value are 7.
10mg DAP-2-DG are dissolved in 0.5mL 0.5molL-1PH7.0 phosphate buffers in, add three carbonyl technetiums-
In 99m midbody solutions.15min is reacted under 95 DEG C of heating conditions.Obtain99mTc(CO)3-DAP-2-DG。
Identified by thin-layer chromatography and HPLC, its radiochemical purity is all higher than 99%.Retention time is in HPLC
10.31min.HPLC conditions are：High performance liquid chromatograph HITACHI (D-2000)；Kromaisl C18 posts 250 × 4.6mm, 5
μmA phases are water (0.1%TFA), and B phases are methanol (0.1%TFA)；Eluting gradient is：0-3min：5% B phases；
3-3.1min：5%~25%B phases；3.1-9min：25%B phases；9-9.1min：25%~34%B phases；9.1-20min：34%
~100%B phases；20-25min：100%B phases；25-25.1 min：100%~5%B phases；25.1-30min：5%B phases.Flow velocity
1mL·min-1。
The preparation of the technetium-99 m labeled complex of the carbonyl of embodiment 3 three
Sodium borohydride 5.5mg, sodium potassium tartrate tetrahydrate 20mg, sodium carbonate 4mg are weighed respectively to be placed in penicillin bottle, are covered
Aluminium lid seals after plug, extracts air with pumping pin, is passed through CO gases 10-20min thereto.Will be from medical99Mo-99mTc is sent out
Eluted in raw device acquisition Sodium Pertechnetate Na [99mTcO4] leacheate 2mL (about 2mCimL-1), add in reaction bulb, shake up molten
30min is reacted after solution under 75 DEG C of metal bath heating conditions, that is, obtains three carbonyl technetium -99m intermediates.Add 1molL-1Salt
Acid, regulation pH value are 7.
To, 2mg DAP-3-DG are dissolved in 0.5mL 0.5molL-1In pH7.0 phosphate buffers, three carbonyl technetiums of addition-
In 99m midbody solutions.15min is reacted under 95 DEG C of heating conditions.Obtain99mTc(CO)3-DAP-3-DG。
Identified by thin-layer chromatography and HPLC, its radiochemical purity is all higher than 99%.Retention time is in HPLC
10.17min.HPLC conditions are：High performance liquid chromatograph HITACHI (D-2000)；Kromaisl C18 posts 250 × 4.6mm, 5
μmA phases are water (0.1%TFA), and B phases are methanol (0.1%TFA)；Eluting gradient is：0-3min：5% B phases；
3-3.1min：5%~25%B phases；3.1-9min：25%B phases；9-9.1min：25%~34%B phases；9.1-20min：34%
~100%B phases；20-25min：100%B phases；25-25.1 min：100%~5%B phases；25.1-30min：5%B phases.Flow velocity
1mL·min-1。
The preparation of the technetium-99 m labeled complex of the carbonyl of embodiment 4 three
Sodium borohydride 5.5mg, sodium potassium tartrate tetrahydrate 20mg, sodium carbonate 4mg are weighed respectively to be placed in penicillin bottle, are covered
Aluminium lid seals after plug, extracts air with pumping pin, is passed through CO gases 10-20min thereto.Will be from medical99Mo-99mTc is sent out
Eluted in raw device acquisition Sodium Pertechnetate Na [99mTcO4] leacheate 2mL (about 2mCimL-1), add in reaction bulb, shake up molten
30min is reacted after solution under 75 DEG C of metal bath heating conditions, that is, obtains three carbonyl technetium -99m intermediates.Add 1molL-1Salt
Acid, regulation pH value are 7.
20mg DAP-5-DG are dissolved in 0.5mL 0.5molL-1 pH7.0 phosphate buffers, three carbonyl technetiums of addition-
In 99m midbody solutions.15min is reacted under 95 DEG C of heating conditions.Obtain99mTc(CO)3-DAP-5-DG。
Identified by thin-layer chromatography and HPLC, its radiochemical purity is all higher than 99%.Retention time is in HPLC
10.45min.HPLC conditions are：High performance liquid chromatograph HITACHI (D-2000)；Kromaisl C18 posts 250 × 4.6mm, 5
μmA phases are water (0.1%TFA), and B phases are methanol (0.1%TFA)；Eluting gradient is：0-3min：5% B phases；
3-3.1min：5%~25%B phases；3.1-9min：25%B phases；9-9.1min：25%~34%B phases；9.1-20min：34%
~100%B phases；20-25min：100%B phases；25-25.1 min：100%~5%B phases；25.1-30min：5%B phases.Flow velocity
1mL·min-1。
Application examples 1
With gained in embodiment 399mTc(CO)3Exemplified by-DAP-3-DG, Bioexperiment process is as follows：
The Kunming small white mouse of 15 lotus S180 tumours is taken, in tail vein injection 0.1mL99mTc(CO)3- DAP-3-DG is (about
0.185MBq).The sacrificed by decapitation after 30,120 and 240min after injection.Take out the heart, liver, lung, kidney, muscle, bone, tumour, blood etc.
Tissue, weigh and its radiocounting is surveyed in gamma counter.Its bio distribution in tumor-bearing mice the results are shown in Table 1.
Table 1.99mTc(CO)3- DAP-3-DG is at Biodistribution in mice data (%ID/g ± sd, n=5)
The result of table 1 shows,99mTc(CO)3- DAP-3-DG after injection can be in tumor region concentration, after injection
30min, it is 1.01%ID/g in the intake of tumor locus, it is relatively low in the intake of non-target organ, and tumour/muscle ratio is reachable
2.05, mainly excreted by kidney tachymetabolism.
4h after injection, tumour/muscle ratio is 2.31, and Clinical practice under equal conditions18F-FDG is in lotus S180
After mice with tumor injection 4h, tumour/muscle ratio is only 0.51.
The Kunming small white mouse of lotus S180 tumours, in tail vein injection 0.1mL99mTc(CO)3- DAP-3-DG is (about
7.4MBq).After isoflurane anesthesia, it is acquired after 30,60 and 120min after injection using SPECT scanners.Lotus knurl
Mouse SPECT imaging results are consistent with bio distribution, and 30min has obvious radioactivity concentration in tumor region after injection.
Its removing in vivo is very fast, mainly enters bladder by renal metabolism, and finally excrete.
Application examples 2
Will99mTc(CO)3- DAP-n-DG be respectively placed in physiological saline, 0.001mol/L cysteine,
In 0.001mol/L histidine, 2% hyclone solution, at 37 DEG C warm bath after 24 hours radiochemicsl purity it is unchanged, explanation99mTc(CO)3- DAP-n-DG has preferable stability.
Above-mentioned application examples explanation, 2, the 3- diaminopropionic acids coupling amino of three carbonyl technetium -99m cores of the invention mark
Glucose coordination compound (99mTc(CO)3- DAP-n-DG) there is high radiochemical purity (being more than 99%), excellent is external steady
It is qualitative, there is preferable intake in tumour, and removing speed in vivo is very fast, can reduce unnecessary radioactive damage.With
Clinical practice18F-FDG can show compared to there is preferable tumour/muscle target to non-target ratio value as new tumour SPECT
As agent.
Unless limited otherwise, term used herein is the implication that those skilled in the art are generally understood that.
Embodiment described in the invention is not used to limit the scope of the invention merely for exemplary purpose,
Those skilled in the art can be made within the scope of the invention various other replacements, changes and improvements, thus, the present invention is unlimited
In above-mentioned embodiment, and only it is defined by the claims.
Claims (11)
1. a kind of Glucosamine derived ligand compound, it has the structure as shown in formula (A),
Wherein ,-(CH2)n- it is straight chain or the structure comprising side chain, n represents 2~8 integer.
2. Glucosamine derived ligand compound according to claim 1, wherein ,-(CH2)n- it is linear chain structure, n tables
Show 2~5 integer.
3. the preparation method of the Glucosamine derived ligand compound of claim 1 or 2, comprises the following steps：
(1) it is that initiation material is reduced to 2- amino -2- t-butylcyanoacetates with 2- oximido -2- t-butylcyanoacetates (I)
(II)；
(2) amino of the 2- amino -2- t-butylcyanoacetates (II) obtained by step (1) is subjected to radical protection and obtains intermediate
Compound (III), and with halogenated carboxylic ester R ' OOC (CH2)nX reacts to obtain midbody compound (IV-n)；
(3) it is amino by the cyano reduction in the midbody compound (IV-n) obtained by step (2) and carries out radical protection and obtain
Midbody compound (V-n)；
(4) hydrolysis of ester group of the midbody compound (V-n) obtained by step (3) is obtained into midbody compound (VI-n)；
(5) midbody compound (VI-n) obtained by step (4) and four acetoxyl group Glucosamines are reacted to obtain intermediate
Compound (VII-n)；
(6) midbody compound (VII-n) obtained by step (5) is subjected to group deprotection, hydrolysis and obtains the aminoglucose
Sugared derived ligand compound (A)；
Synthetic route is as follows：
4. preparation method according to claim 3, wherein, the halogenated carboxylic ester R ' OOC (CH2)nIn X, X represents F, Cl
Or Br, R ' represent C1~C10 straight or branched alkyl.
5. preparation method according to claim 4, wherein, R ' represents C1~C5 straight or branched alkyl.
6. a kind of technetium-99 m labeled complex of three carbonyl, it has the structure as shown in formula (B),
Wherein ,-(CH2)n- it is straight chain or the structure comprising side chain, n represents 2~8 integer, and M is represented99mTc。
7. complex according to claim 6, wherein ,-(CH2)n- it is linear chain structure, n represents 2~5 integer.
8. the preparation method of the technetium-99 m labeled complex of three carbonyl of claim 6 or 7, comprises the following steps：
(1) prepare three carbonyl technetium -99m intermediate hydrated ions fac- [99mTc(CO)3(H2O)3]+；
(2) by the fac- of gained [99mTc(CO)3(H2O)3]+With the Glucosamine derived ligand compound of claim 1 or 2
The complex is made in reaction.
9. preparation method according to claim 8, wherein, in the step (2), fac- [99mTc(CO)3(H2O)3]+Will with right
Ask the 1 or 2 Glucosamine derived ligand compounds to be heated to 70~100 DEG C of 5~30min of reaction in neutral conditions to be made
The complex.
10. preparation method according to claim 9, wherein, the neutrallty condition is the phosphate buffer that pH value is 7.
11. the complex described in claim 6 or 7 is used for the purposes for preparing tumour SPECT developers.
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| CN102146098A (en) * | 2011-03-02 | 2011-08-10 | 北京师范大学 | Preparation method and application of 99mTc labeled D-glucose coordination compound |
| CN102977174A (en) * | 2012-12-19 | 2013-03-20 | 北京师范大学 | 99mTc(CO)3 nuclear-labeled macrocyclic polyamine triazole ring glucose-based complex as well as preparation and application of complex |
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| CN102146098A (en) * | 2011-03-02 | 2011-08-10 | 北京师范大学 | Preparation method and application of 99mTc labeled D-glucose coordination compound |
| CN102977174A (en) * | 2012-12-19 | 2013-03-20 | 北京师范大学 | 99mTc(CO)3 nuclear-labeled macrocyclic polyamine triazole ring glucose-based complex as well as preparation and application of complex |
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| Glucosamine conjugates bearing N, N, O-donors: potential imaging agents utilizing the [M (CO) 3]+ core (M= Re, Tc);Bowen M L, Lim N C, Ewart C B, 等;《Dalton Transactions》;20090925(第42期);第9216-9227页 * |
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