CN108144072A - For diagnosing the radiopharmaceutical of agglutinin receptor height expression tumour - Google Patents
For diagnosing the radiopharmaceutical of agglutinin receptor height expression tumour Download PDFInfo
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- CN108144072A CN108144072A CN201711450692.XA CN201711450692A CN108144072A CN 108144072 A CN108144072 A CN 108144072A CN 201711450692 A CN201711450692 A CN 201711450692A CN 108144072 A CN108144072 A CN 108144072A
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- A61K51/00—Preparations containing radioactive substances for use in therapy or testing in vivo
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- C07D495/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms in which the condensed system contains two hetero rings
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Abstract
A kind of radiopharmaceutical for being used to diagnose agglutinin receptor height expression tumour, the radiopharmaceutical is a kind of99mThe label complex of Tc, with core sequestration ligand, the ligand is using L1 as ligand precursor, and the drug is as radiation diagnostic imaging agent, for diagnosing Malignancy and entity tumor, particularly for the diagnosis of colorectal cancer.
Description
Technical field
The present invention relates to a kind of diagnosing tumor based on agglutinin receptor pre-targeting strategy radiopharmaceutical more particularly to
A kind of radiopharmaceutical for the colorectal cancer for being used to diagnose agglutinin receptor mediation.
Background technology
Agglutinin (Lectin) is a kind of protein that can be specifically bound with glycosyl in sugar compounds in life entity,
Its studied history is more than more than 100 years.The last century 90's, scientists find to be aggregated in many gastroenteric tumors
Plain expression quantity is significantly larger than Tumor-surrounding tissue.Such as 21 kinds of colorectal cancer cell systems are once carried out molecule life by Ohannesian et al.
Object credit is analysed, and discovery has 20 kinds to express agglutinin.Therefore, agglutinin is undoubtedly a good medicine for colorectal cancer
Object action target spot can be used for the diagnose and treat of tumour.
Avidin (Avidin) is extracted from egg white, and 66-69kDa of molecular weight is a kind of high glycosylation, positively charged
Protein (pI=10.5), glucose and mannose residue containing end N- acetylations.These saccharide residues can be by agglutinin spy
Opposite sex identification, and the combination that high-affinity occurs forms Avidin-Lectin compounds.In vivo, Avidin can be special
The Lectin of the combination tumour cell height expression of the opposite sex, the Avidin not combined with Lectin can reenter blood circulation, quilt
Liver, which absorbs and passes through liver and gall, to be metabolized out in vitro.Therefore, Avidin can be as the ligands specific of Lectin, for being surveyed in physical examination
The targeted molecular or idiosyncratic carrier of Lectin expressions.
Nuclear medicine molecular image technology, such as positron emission fault (PET) and single photon emission tomography (SPECT) skill
Art compared with traditional detection means, has many advantages, such as high sensitivity, noninvasive, quick, real-time, in early diagnosis of tumor
Aspect possesses huge advantage.Correlative study reports that Lectin (agglutinin) high receptor is expressed in the surface of kinds of tumor cells,
Especially in gastroenteric tumor, such as colorectal cancer, expression degree are proportionate with grade malignancy.Therefore, for Lectin
For the tumour of high receptor expression, it is undoubtedly a good radiopharmaceutical action target spot.At present, for Lectin's
For nuclear medicine molecular probe using Avidin as design core, but such probe mostly molecular weight is excessive, structure is excessively complicated, unfavorable
It is prepared in following medicine boxization, secondly, all containing amino acid such as lysine in probe structure, this can cause probe to be expressed with height
Megalin albumen in distal renal tubular position is combined, and probe is caused to absorb the excessive and residence time in renal tract
Long, tumour/kidney ratio is relatively low, and large effect is produced to the diagnosis for being located at abdominal cavity region tumors.In addition, particularly for
It is the research of target spot mostly based on colorectal cancer abdominal cavity dissemination model using Lectin, this class model is with facing for colorectal cancer
Bed actual conditions have certain gap, can not reflect the true medicine of drug function in an acting capacity of for.
Invention content
In order to improve the above problem of the existing technology, the present invention provides a kind of99mThe radiolabeled complexes of Tc,
The complex has core sequestration ligand, and the ligand is using following formula L1 or its stereoisomer form as its ligand precursor：
Wherein：
- L- is selected from linking arm as follows：
The C that R is independently selected from H, is optionally optionally substituted by a hydroxyl group1-6Alkyl, C1-6Alkoxy, halogen ,-NH2；
Integers of the n for 2-8, such as 2,3,4,5,6,7,8；
Integers of the s for 0-6, such as 0,1,2,3,4,5,6；
R1For H ,-SO3H ,-COOH or described sulfonic acid, carboxylic acid pharmaceutically acceptable salt or ester.
According to the present invention, for above-mentioned ligand precursor formula L1 when forming label complex, C=N double bonds break to form N=N
Double bond is coordinated with technetium.L1 is the ligand precursor based on buzane niacinamide (HYNIC) chelating agent.Known HYNIC is a kind of
Effective Tc chelating agents are participated in since HYNIC can only provide 1-2 coordination atom99mTc is complexed, therefore in addition to the present invention is above-mentioned
Outside the core sequestration ligand of definition, can also have the synergy modes L2 and/or L3 of auxiliary.It, can be with as synergy modes L2, L3
It is identical or different, be it is known in the state of the art can conduct99mThose of Tc ligands, wherein common synergy modes L2, L3 includes
Water-soluble phosphine (such as three sulfonate sodium TPPTS of triphenylphosphine), N- tri- (methylol) methylglycine (Tricine), N- bis-
(ethoxy) glycine, gluceptate, ethylenediamine-N, N '-diacetate esters (EDDA), 3- benzoyl pyridines (BP), pyridine-
2- azo-p- dimethylanilines (PADA) etc..
It is described according to the present invention99mTc includes the technetium radioisotope of arbitrary redox state, for example, its+6 ,+5 ,+4,
+ 3 ,+2 ,+1 valence state, preferably99mTc (III),99mTc (IV),99mTc(VII)。
According to the present invention, in the formula L1, R1 is the substituent group on its phenyl ring, and position can be the o-, m-, right of hydrazone key
Position.
Preferably, it is of the invention99mIn the radiolabeled complexes of Tc：
Linking arm L in L1 is selected from formula d structures；Preferably wherein s be 2, n 3；
R1For-SO3H；
Synergy modes may be the same or different, independently selected from：Three N- tri- (methylol) methylglycine, triphenylphosphine sulphurs
Acid sodium-salt.
As example, core sequestration ligand precursor formula L1 of the invention is following formula or its stereoisomer：
As example, label complex of the invention (is abbreviated as99mTc-HYNIC-PEG3- B) be：
The label complex of the present invention as above since the combination of ligand acts synergistically, is kept in the solution and in vivo
Stablize.The complex in urine sample by being recovered to the complete complex after intravenous injection, without finding physics and chemistry degradation
Object, this is enough the stability for illustrating the complex.
The present invention also provides described99mThe preparation method of the radiolabeled complexes of Tc, includes the following steps：
(1) it after the solution of the solution of ligand precursor L1, the solution of L2 and/or L3 is mixed, is added in into mixed solution99mTc sources, reaction produce the label complex；Or ligand precursor L1, ligand L 2 and/or L3 are dissolved in buffer solution,
It is added in into mixed solution99mTc sources, reaction produce the label complex；
Preferably, the ligand precursor formula L1 is obtained by following steps (2) coupling, it will be appreciated by those skilled in the art that
Wherein described L is the structure except the linking arm L removings-NH- during the above-mentioned L1 of the present invention is defined：
(2)
The present invention99mThe above-mentioned preparation method of the radiolabeled complexes of Tc, is known in the art method.
Preparation in accordance with the present invention in the labeling method of step (1), using known addition or is not added with SnCl2
Labelling method, wherein reaction temperature according to specific reaction raw materials and whether need add in SnCl2It can difference, such as 15-100
DEG C, it is then generally heated if you need to heat using water-bath or air bath, the reaction time is usually dozens of minutes to a few houres, preferably
20-60min.The precursor of ligand L 1 can use deionized water dissolving, and synergy modes L2, L3 can be selected accordingly according to property
Buffer solution dissolves, such as succinate buffer may be selected in TPPTS, Tricine.Described99mTc sources are usually its salt, such as
Na99mTcO4。
Preparation in accordance with the present invention, usually after the label of step (1), such as with known Radio-HPLC
Method, Sep-Pak reverse-phase chromatographic columns, Sephadex G-25 columns, YMC-Pack ODS-A C18Analytical column etc. calculates its mark after purification
Note rate.
Preparation in accordance with the present invention, in the coupling reaction of step (2), the raw material SBZ-HYNIC is known substance
Matter, structure are：Another raw material is commercially available, such as BioMatrik (is won in Jiaxing
It is beautiful) series of products of company.R in the present invention1The coupling agent of the step of being defined for other (2) can be prepared by raw material SBZ-HYNIC
It obtains, such as sulfonic esterification, it can also be byIt is anti-that hydrazone key occurs with the aldehyde with carboxylic acid phenyl
It should.
The coupling reaction carries out in the mixed solution of reactant, the optional dimethylformamide freely (DMF) of solvent, N,
N- dimethylacetylamides (DMAC), dimethyl sulfoxide (DMSO) (DMSO) etc.；PH value in reaction can be adjusted between 8-9, such as 8.5；Temperature
It can be 15-50 DEG C, be preferably close to room temperature, such as 25 DEG C；The time of the reaction can be 2-30 hours, in reaction process,
Reaction process can be detected with HPLC and receive sample.
Can the use of mobile phase be volume ratio in 0-5min it be 100 when using high-efficient liquid phase color time spectrum:0 mobile phase
A：Mobile phase B；The use of mobile phase is later volume ratio to 35min or so it is 20:80 mobile phase A:Mobile phase B；Mobile phase A can
Think the deionized water containing 0.05% trifluoroacetic acid, Mobile phase B can be the acetonitrile containing 0.05% trifluoroacetic acid.
The present invention it has been investigated that, the present invention99mThe radiolabeled complexes of Tc have specific binding with Avidin,
And Avidin can specificity combination tumour cell height expression Lectin receptors, therefore, label complex of the invention can
As nuclear medicine image molecular probe of the Lectin receptors after Avidin pre-targeting.
Therefore, the present invention also provides a kind of nuclear medicine molecular probes, as defined above for the present invention99mTc radioactivity marks
Remember complex.
The present invention also provides a kind of pharmaceutical composition, the composition includes a effective amount of above-mentioned label complex.
Preferably, pharmaceutical composition of the invention is a kind of diagnostic medicine, is a kind of radiation diagnostic imaging agent.It is described to put
Penetrating property diagnosing developing agent can be used in combination with Avidin, after subject Avidin is given, be directly applied to individual, lead to
It crosses nuclear medical imaging device and detects gamma (γ) ray that the complex applied to subject is emitted, obtain visualising image
Diagnosis.Preferably, pharmaceutical composition of the invention is a kind of injectable formulation, and it includes above-mentioned label complex and injectables
Carrier.It is preferred that the developer refers to the developer as SPECT.
According to the present invention, described pharmaceutical composition may include described99mOne kind of the radiolabeled complexes of Tc, two kinds
Or more kind.
The present invention also provides a kind of drug system, the system includes a effective amount of99mThe radiolabeled complexes of Tc and
Avidin。
Preferably, the drug system is a kind of kit, and label complex of the invention and Avidin are divided in described
In kit.
In addition, the present invention also provides it is a kind of it is convenient prepare it is above-mentioned99mTc marks the medicine box of complex, and the medicine box, which contains, matches
The lyophilized preparation of the mixture of body precursor L1, ligand L 2 and/or L3.Preferably, before the medicine box can be by that will contain ligand
The buffer solution of body L1, ligand L 2 and/or L3 are lyophilized to be prepared into cillin bottle.In each label cooperation for carrying out the present invention
When prepared by object, the medicine box prepared only need to be taken out, adds in corresponding radioactive activity99mTc sources, such as Na99mTcO4, in 80-
120 DEG C, preferably 100 DEG C of water-baths heat 10-60 minutes, preferably 20-30 minutes, you can the label complex of the present invention is made.More
Preferably, the ligand precursor 15-25 μ g of above-mentioned formula L1, such as 20 μ g are weighed；Weigh N- tri- (methylol) methylglycine
(Tricine) 7.5-12.5mg, such as 10mg；Three sulfonate sodium (TPPTS) 3.75-6.25mg of triphenylphosphine are weighed, such as
5mg；It is a certain amount of to weigh succinic acid；Extract water is a certain amount of；More than substance is placed in cillin bottle and adjusts pH using sodium hydroxide
Value is lyophilized after filtering to 5.5 and obtains medicine box.
The present invention also provides the purposes of the label complex in medicine preparation.According to the present invention, the drug is made
For developer for diagnosing Malignancy and entity tumor, for example, blood, liver, intestines, kidney, stomach, spleen, lung, muscle,
The malignant tumour at the positions such as bone.It is preferred that the cancer of those agglutinins Lectin receptors height expression, preferably gastrointestinal cancer, such as
Colorectal cancer and the hepatic metastases by its initiation.
The present invention also provides a kind of methods for diagnosing hematological system and solid malignant, particularly colorectal cancer, will have
The label complex of the present invention of effect amount is applied to the individual of this demand.
According to the present invention, the individual can be mammal, such as mankind.
Advantageous effect
The present invention it has been investigated that, efficient specificity occurs for label complex of the invention and ligand Avidin Avidin
With reference to, and Avidin can with the agglutinin Lectin specific bindings of expression high in tumour, and affinity higher than polypeptide with by
Affinity between body, is slightly inferior between Ag-Ab, therefore the label complex of the present invention is enriched with by we using Avidin
In tumor locus.In addition, we also demonstrate that the label complex can be imaged as developer for the SPECT of tumour,
By imaging demonstrate the label complex can specificity concentrate in tumor locus, there is very strong in situ tumor to be imaged
Ability, and property is superior in vivo.More, it is surprising that the distribution of the label complex of the present invention in vivo has very
High tumour/non-tumour ratio.
Colorectal cancer original site be concentrated mainly on it is intraperitoneal, and the present invention99mTc marks complex to be visited as molecule
Needle because molecular weight is small, avoids the deficiency that imaging abdominal cavity background is high in the prior art.In addition, the present invention99mTc label cooperations
For object not only in common colorectal cancer abdominal cavity dissemination model, tumor locus uptake ratio is high, and renal tract uptake ratio is substantially reduced,
With excellent tumour/kidney ratio, and the tumor imaging in colorectal cancer model and liver metastasis model is induced, it is also the same to have
There is good tumour/kidney ratio, this is of great significance for the diagnosis of colorectal cancer.
Therefore, label complex of the invention, pre-targeting system include but not limited to advantages below：
(1) label complex of the invention is combined rapidly with Avidin, and mixing 30min at room temperature can tie mutually
It closes；
(2) pre-targeting system and tumour cell binding force of the invention are strong, such as are with the Percentage bound of colorectal cancer cell
48.23 ± 3.16%；
(3) pre-targeting systemic characteristic of the invention is strong, and pre-targeting system normal tissue organ of the present invention is without significantly taking the photograph
It takes, but there is high uptake ratio in tumor locus；
(4) property is superior in pre-targeting system body of the invention, and distribution in vivo has excellent tumour/non-tumour ratio
Value, especially tumour/kidney ratio.
Term defines and explanation
Unless otherwise defined, the connotation that all scientific and technical terminologies have herein and claim theme fields technology
The normally understood connotation of personnel is identical.Unless otherwise indicated, all patents, patent application, the public material being cited in full text herein
It is integrally incorporated by reference herein.If there are multiple definition to term herein, it is subject to the definition of this chapter.
Numberical range described herein, when the numberical range is defined as " integer ", it should be understood that describe this
Two endpoints of range and each integer in the range of this.For example, " 0~10 integer " should be understood to describe 0,1,
2nd, 3,4,5,6,7,8,9 and 10 each integer.When the numberical range is defined as " counting ", it should be understood that describe this
Two endpoints of range, each integer in the range of this and each decimal in the range of this.For example, " 0~10 number "
It should be understood to not only describe 0,1,2,3,4,5,6,7,8,9 and 10 each integer, also at least describe wherein each
A integer respectively with 0.1,0.2,0.3,0.4,0.5,0.6,0.7,0.8,0.9 and.
Term " optionally/arbitrary " or " optionally/arbitrarily " refer to the event then described or situation may occur or can
It can not occur, which includes the event or situation occurs and the event or situation does not occur.
Term " C1-6 alkyl " is interpreted as the preferred direct-connected or branch saturation monovalent hydrocarbon for representing to have 1~6 carbon atom
Base represents the direct-connected or branch saturation monovalent hydrocarbon with 1,2,3,4,5,6 carbon atom.The alkyl is such as methyl, second
Base, propyl, butyl, amyl, hexyl, isopropyl, isobutyl group, sec-butyl, tertiary butyl, isopentyl, 2- methyl butyls, 1- methyl fourths
Base, 1- ethyl propyls, 1,2- dimethyl propyls, neopentyl, 1,1- dimethyl propyls, 4- methyl amyls, 3- methyl amyls, 2- first
Base amyl, 1- methyl amyls, 2- ethyl-butyls, 1- ethyl-butyls, 3,3- dimethylbutyls, 2,2- dimethylbutyls, 1,1- bis-
Methyl butyl, 2,3- dimethylbutyls, 1,3- dimethylbutyls or 1,2- dimethylbutyls etc. or their isomers.
Alkyl in term " C1-6 alkoxies " has the meaning identical with above-mentioned " C1-6 alkyl ".
Term " halogen " refers to F, Cl, Br, I.
Term " sulfonic group, carboxylic acid group " is understood to mean that the univalent perssad-SO such as lower structure3H ,-COOH structures.They
Ester group refer to carboxylic acid group, sulfonic group formed Arrcostab, acid imide base ester, the definition of alkyl for example above-mentioned " C1-6 alkyl ".
Term " acid imide " is interpreted as the structure that general formula is R-C (O)-N (R)-C (O)-R, such as phthalyl Asia
Amine, succinimide, N-bromosuccinimide, glutarimide, maleimide etc..
Term " stereoisomer " refers to the isomers as caused by the spatially arrangement mode difference of atom in molecule.This
Invention L1 ligands contain asymmetric or chiral centre, and accordingly, there exist different stereoisomeric forms in any ratio.All stereochemical structures of L1 and mixed
It is the same to close object, including racemic mixture, as the part applied at present.
Term " pharmaceutically acceptable salt " refers to remain the free acid of appointed compound and the biopotency of free alkali,
And there is no the salt of ill-effect on biology or other aspects.The application ligand precursor L1, which is further included, can pharmaceutically receive
Sulfonate or carboxylate, such as sodium salt, sylvite, calcium salt etc..Pharmaceutically acceptable salt refers to the sulphur in parent compound
Acidic group or carboxylic acid group are converted into the form of salt.The application pharmaceutically acceptable salt can be synthesized by parent compound, i.e. parent
Sulfonic group or carboxylic acid group in compound are reacted with the alkali of 1-4 equivalents in a solvent system.Suitable salt is enumerated
Remingtong’s Pharmaceutical Scicences,17thed.,Mack Publishing Company,Easton,
Pa., 1985, p.1418 with Journal of Pharmaceutical Science, in 66,2 (1977), such as sodium salt.
Description of the drawings
Fig. 1：HYNIC-PEG in embodiment 13The mass spectral analysis figure of-B；
Fig. 2：In embodiment 199mTc-HYNIC-PEG3The Radio-HPLC analysis charts of-B；
Fig. 3：Embodiment 299mTc-HYNIC-PEG3- B/Avidin mixed liquors with individually99mTc-HYNIC-PEG3- B is in phase
With the elution curve in PD-10 desalting columns；
Fig. 4:Fig. 4 a of embodiment 3 are the streaming of FITC-Avidin, FITC-neutrAvidin and LS180-Luci cell
Cell results figure；Fig. 4 b scheme to be quantitative, * * * P<0.001；
Fig. 5：Embodiment 499mTc-HYNIC-PEG3Biodistribution data figure (ns of-the B in LS180 belly cavity tumor models
=4).
Fig. 6：Embodiment 499mTc-HYNIC-PEG3- B in different time points divide by the quantitative of tumour/kidney ratio (n=4)
Analysis figure.
Fig. 7：The AOM/DSS of embodiment 5 induces colorectal cancer Animal Model timeline；
Fig. 8：The AOM/DSS of embodiment 5 induces model mouse and the Colon and rectum white light photo with batch normal control mouse；
Fig. 9：The AOM/DSS of embodiment 5 induces model mouse (a) and is dyed with the intestinal tissue H＆E with batch normal control mouse (b)
Figure；
Figure 10：Embodiment 599mTc-HYNIC-PEG3- B is induced in normal rat (normal mice) and AOM/DSS to be tied
SPECT/CT images in carcinoma of the rectum model mouse (induced model) (white arrow meaning is tumour)；
Figure 11：The induction model mouse dissection white light photo (a, b) of embodiment 5, the SPECT/CT images (c) of intestinal tissue
With two at colorectal carcinoma H＆E colored graphs (d, e) (at white arrow meaning be tumour) comparison diagram；
Figure 12：Embodiment 699mTc-HYNIC-PEG3- B is in LS180-Luci colorectal cancer liver metastasis models
SPECT/CT schemes (a) and archebiosis light (BLI) imaging figure (b) (being tumour at white arrow meaning)；
Specific embodiment
Marker of the present invention and its preparation method and application is done below in conjunction with specific embodiment further detailed
It describes in detail bright.It should be appreciated that the following example is merely illustrative the ground description and interpretation present invention, and it is not necessarily to be construed as to the present invention
The limitation of protection domain.All technologies realized based on the above of the present invention are encompassed by the range the present invention is directed to protection
It is interior.
Unless otherwise indicated, the raw materials and reagents used in following embodiment are commercial goods or can be by
It is prepared by perception method.
The marker of 1 formula III of embodiment (is abbreviated as99mTc-HYNIC-PEG3- B) preparation
HYNIC-PEG3The synthesis of-B
HPLC methods：YMC-Pack ODS-A C18 analytical columns are prepared using Agilent 1260Infinity hplc devices
(250 × 4.6mml.D.S-5 μm, 12nm)/semi-preparative column (250 × 10mml.D.S-5 μm, 12nm), gradient elution time is
42min, flow velocity are 1mL/min (analysis) or 4mL/min (receiving sample), and flowing A phases are deionized water (containing 0.05%TFA), flow B
It is mutually acetonitrile (containing 0.05%TFA).Eluting gradient is：100%A and 100%A and 0%B, 35min when 0%B, 5min during 0min
When 20%A and 100%A and 0%B when 80%B, 42min.
HYNIC-PEG3- B passes through NH2-PEG3The amino of-B (structure is shown in following synthetic route) and the hydroxyl of SBZ-HYNIC
Succinimide activated ester is coupled to obtain, and synthetic route is as follows.Accurately weigh 4mg NH2-PEG3(9.5 μm of oL, are dissolved in-B
200 μ L DMF) it is placed in 1.5mL EP pipes；Accurately SBZ-HYNIC 1.8mg (4.3 μm of oL are dissolved in 300 μ L DMF) are weighed to be placed in
In EP pipes；Mix NH2-PEG3B solution and SBZ-HYNIC solution, add in micro DIEA (about 1.5 μ L) by reacting solution pH value tune
System 8.5 to 9.Room temperature concussion reaction 2h.Reaction process is detected using HPLC, gradient used is shown in above-mentioned HPLC methods.Through HPLC points
Analysis is it is found that under the gradient, NH2-PEG3The retention time of-B is 6.25min, and the retention time of SBZ-HYNIC is 10.25min.
After reaction carries out 2 hours, HPLC detection reactions find occur new ultraviolet absorption peak at 13.90min, and with the reaction time
Extension gradually increase, and raw material peak is gradually reduced.After reaction 24 hours, raw material ultraviolet absorption peak completely disappears, and use is identical
HPLC methods carry out receipts sample, collect the substance that ultraviolet absorption peak is located at 13.90min, are put into freeze dryer freeze-drying, obtain white powder
Last shape substance.It is dissociated through ground substance assistant laser and absorbs ionization time of flight mass spectrometry (MALDI-TOF-MS) identification, at 13.90min
For product HYNIC-PEG3-B：M/z=722.34 ([M+H]+), C31H43N7O9S2Theoretical molecular weight is 721.84.Attached drawing 1 is production
The mass spectral analysis figure of object.
99mTc-HYNIC-PEG3The preparation of-B
Using TPPTS and Tricine one-step method to HYNIC-PEG3- B is carried out99mThe label of Tc.Detailed step is as follows：Essence
Really weigh 20 μ g HYNIC-PEG3- B (1 μ g/ μ L are dissolved in deionized water), (100 μ g/ μ L are dissolved in 25mM ambers to 5mg TPPTS
In acid buffer, pH=5.5) and 10mg Tricine (100 μ g/ μ L are dissolved in 25mM succinate buffers, pH=5.5) additions
Into 1.5mL EP pipes.150 μ L Na are added in into mixed liquor99mTcO4(about 1100MBq).It is placed in air bath and is heated to 100
DEG C, standing treats that it is cooled to room temperature after reacting 20min.After label, mark rate is detected using Radio-HPLC>98%, nothing
Need to purify can be used for cell or zoopery, Radio-HPLC methods and HYNIC-PEG3- B synthesizes HPLC analysis method phases
Together.Attached drawing 2 is the Radio-HPLC analysis charts of the marker.
It, will using sterile saline after Radio-HPLC mark rates detect99mTc-HYNIC-PEG3- B is diluted to
Suitable concentration is accordingly to be tested.After interior animal experiment is filtered degerming using 0.22 μm of filter membrane to marker, then root
Requirement is diluted to the parenteral solution of corresponding radioactive concentration according to the experiment, is injected into animal body and is tested.
Embodiment 299mTc-HYNIC-PEG3- B is combined the measure of activity with Avidin in vitro
200 μ g Avidin are taken, are dissolved in 100 μ L PBS (pH=7.4), add in 37MBq99mTc-HYNIC-PEG3- B is mixed
It is even, it is placed at room temperature for 0.5h.Pass through PD-10 desalting columns pair99mTc-HYNIC-PEG3- B/Avidin mixed liquors carry out separation analysis,
It is described in detail below.First, 5-10 is carried out to PD-10 desalting columns using PBS solution all over cleaning, removes anti-corrosion remaining in column
Liquid.After cleaning, residue PBS in column is discharged, then mixed liquor is added in column and after it is submerged in column, takes PBS pairs
PD-10 desalting columns carry out in batches repeatedly elution (200 μ L every time, totally 18 times, 3.6mL), and leacheate is respectively placed in 1.5mL
In EP pipes (200 μ L/ pipes), each EP pipes are finally respectively placed in radiological measuring borehole measurement radioactive activity and are recorded.
It treats99mTc-HYNIC-PEG3It is with 10mL PBS that PD-10 desalting columns is clear after-B/Avidin mixed liquors elute
Wash clean then applies same procedure to individual in the PD-10 desalting columns99mTc-HYNIC-PEG3- B is eluted.
The data obtained application software Prism 5.0 carries out the description of elution curve respectively in this experiment.Attached drawing 3 is bent for elution
Line.
We by Avidin with99mTc-HYNIC-PEG3- B is after mixed at room temperature 30min, using PD-10 desalting columns to this
Mixed liquor and individually99mTc-HYNIC-PEG3- B is eluted respectively, calculates respective elution curve.According to PD-10 desalinations
Column principle is it is found that the big substance of molecular weight can be eluted at first, the required leacheate small volume used, and molecular weight is smaller
Substance can be eluted by after, then required elution volume is larger.As a result as shown in Figure 3,99mTc-HYNIC-PEG3-B/
The elution volume section of Avidin mixed liquors is 0.8mL-1.6mL, and individual99mTc-HYNIC-PEG3B solution is identical
The elution section of PD-10 desalting columns be 1.4mL-3.0mL, it can be seen that, Avidin with99mTc-HYNIC-PEG3- B is at room temperature
Mixing 30min can be combined with each other, and cause required elution volume significantly greater than individual99mTc-HYNIC-PEG3-B.The reality
Verification understands the radionuclide obtained as the ligand described in L1 of the present invention99mThe marker of Tc has to be specifically bound with Avidin
Ability, can be used for subsequent zoopery.
3 flow cytometry of embodiment
Cell culture
LS180-Luci colorectal cancer cells are incubated at that (fetal calf serum is before use in 56 DEG C of water containing 10% fetal calf serum
Carry out inactivating for 30 minutes in bath) DMEM (high sugar) culture medium in.Cell is placed in moisture-saturated in incubation, and temperature is
37 DEG C, while maintain 5%CO2In the constant temperature cell incubator of concentration.
Cell culture is once passed on for 2 to 3 days, and cell passage needs to be operated in aseptic operating platform.During passage
Old culture medium in culture bottle is discarded first, and cell is rinsed one time with sterile PBS, adds in 0.25% in right amount containing EDTA later
Trypsase cell is digested, place 2 minutes after, reject trypsin solution.New culture is added in into culture bottle
Cell is blown and beaten from culture bottle inner wall with sterile dropper after base, and piping and druming is uniform in the medium.Cell is added in new
Culture bottle, and cultivated under optimum conditions.
The synthesis of FITC-Avidin/FITC-neutrAvidin
Accurately weighing Avidin 2mg, (3 μm of ol are dissolved in the Na of 200 μ L pH=8.52HPO4) be placed in 1.5mL EP pipes；
Accurately 1mg NHS-FITC are weighed to be dissolved in 50 μ L DMSO；The two is uniformly mixed, 4 DEG C, after reacting 8h, through PD-10 desalting columns
It is purified.PD-10 desalination column purification steps are：PD-10 columns PBS is rinsed 5-10 times, until after PBS is not present in column,
1mL sample F ITC-Avidin to be purified are added in, until after sample is all flowed into column, 1.5mL PBS is added in, exists using EP pipes
The sample of pillar received down after purification.
Sample F ITC-Avidin after purification is subjected to ultraviolet specrophotometer measure.Respectively determination sample in 280nm and
Absorption value at 490nm, after being computed, general chain is connected to 5 FITC molecules on 1 Avidin molecule, and concentration is about 1mg/mL.
The preparation process of FITC-neutrAvidin is same as above.
Flow cytometry
By 1 × 107LS180-Luci colorectal cancer cells are inoculated in culture dish, and DMEM (high sugar) culture medium is incubated overnight
It is adherent to cell.After cell is adherent, culture medium is discarded, ice PBS is cleaned 3 times.NeutrAvidin is a kind of change of Avidin
Body does not have the ability combined with Lectin, therefore this experiment chooses neutrAvidin as control.Experiment is divided into two groups,
FTIC-Avidin incubations group and FITC-neutrAvidin control groups.Divide in the adherent culture dish for there are LS180-Luci cells
It Jia Ru not 10mL (10 μ g/mL) FITC-Avidin or FITC-neutrAvidin incubation 96h.Then discard Incubating Solution, ice PBS
Cell dissociation, is placed in 1.5mL centrifuge tubes by cleaning 3 times using 0.05% pancreatin containing EDTA, 1000 revs/min of centrifugations,
It is cleaned after centrifugation using ice PBS, in triplicate.
After ice PBS is cleaned, each group is separately added into 1mL PBS and cell is resuspended, and sieving moves into streaming pipe, thin using streaming
Born of the same parents' instrument (Becton Dickinson, Germany) is analyzed.Flow cytometry results are shown in attached drawing 4.
Such as attached drawing 4b, under identical incubation conditions, the Percentage bound of FITC-Avidin and LS180-Luci cells is
48.23 ± 3.16%, and the Percentage bound of FITC-neutrAvidin and LS180-Luci cells are only 1.47 ± 0.56%, hence it is evident that
Less than FITC-Avidin.NeutrAvidin is the variant agent of Avidin, and neutrAvidin lacks saccharification compared with Avidin
Structure is learned, does not have the ability for having and being combined with Lectin.Therefore, this experiment is right by the use of FITC-neutrAvidin as feminine gender
According to, it was demonstrated that FITC-Avidin can be combined by selectively targeted Lectin receptors with LS180-Luci tumour cells.
The marker of 4 present invention of embodiment99mTc-HYNIC-PEG3The biodistribution experiments of-B
Animal Model
LS180 human colon cancer cells are purchased from ATCC (Manassas, VA).LS180 is incubated at the DMEM high sugar containing 10%FBS
Cell culture medium.Cell is at 37 DEG C, containing 5%CO2Incubator in routine passage culture.4-5 week old female BAl BIc/c nude mices purchase
From experimental animal portion of Department Of Medicine, Peking University, every in intraperitoneal inoculation 1 × 106A LS180 tumour cells for bio distribution and
Imaging, all zoopery operations follow Department Of Medicine, Peking University animal and use regulation.After tumor cell inoculation 7 days, tumour
Model can be used for biodistribution experiments.
Biodistribution experiments
Lotus LS180 tumour nude mices weight in 20-25g, is randomly divided into four groups, every group 4.Every nude mice is through abdominal cavity
Inject 200 μ g pre-targeting drugs Avidin (being dissolved in 100 μ L physiological saline).After pre-targeting four hours, four groups of mouse are through tail vein
Inject 222kBq's (6 μ Ci)99mTc-HYNIC-PEG3- B is to evaluate its biodistribution characteristics in vivo.After injection 0.5h,
Animal is put to death after 1h, 2h, 4h, takes blood and main organs, weigh and measures radioactivity cpm countings, is counted after decay correction
Calculate the percentage injection dosage (%ID/g) of per gram of tissue.Bio distribution result be expressed as average value ± standard deviation (means ±
SD, n=4).Experimental result is shown in attached drawing 5 and attached drawings 6.Originally it is demonstrated experimentally that after Avidin pre-targeting,99mTc-HYNIC-
PEG3- B has very high tumour/non-tumour ratio, and quantitative analysis in belly cavity tumor99mTc-HYNIC-PEG3- B exists
Different time points tumour/kidney ratio (n=4) that LS180 colorectal cancers abdominal cavity is sent out in tumor model is respectively 0.5 hour
It is (3.78 ± 0.86) for (0.56 ± 0.15), 1 hour, be within 2 hours (6.99 ± 2.61), is within 4 hours (1.47 ± 0.16), it can
See with very high tumour/kidney ratio.
Embodiment 5AOM/DSS induces the SPECT/CT of colorectal cancer mouse
AOM/DSS induces colorectal cancer Establishment of mouse model
AOM/DSS induces colorectal cancer Animal Model and refers to pertinent literature[1,2]Settling time line as shown in Figure 7,
Process is simply described as follows：20 4-5 week old male Balb/c small white mouses are chosen, in intraperitoneal injection 10mg/kg AOM injections in 0 day
Liquid then feeds 2.5%DSS drinking-water 7 days (- 7 days 0 day), is changed to normal water later 14 days (- 21 days 7 days), is so far 1
Cycle, repeats this cycle 3 times, until the 80th day, part mouse is taken to put to death, dissect and observe whether Colon and rectum position has tumour to deposit
It is taking pictures and is taking tumor tissues, fixed using 4% paraformaldehyde, for paraffin section, carrying out H＆E staining analysis.
Meanwhile same batch of 10 4-5 week old male Balb/c small white mouses of raising are as a control group.In 0 day, physiological saline is utilized
It is injected intraperitoneally instead of AOM parenteral solutions, subsequent normal water, until 80 days, part mouse is taken to put to death, dissect and observe Colon and rectum
Position takes pictures and takes intestinal tissue, fixed using 4% paraformaldehyde, for paraffin section, carries out H＆E staining analysis.
It is a kind of tumor model of foundation on inflammatory basis that AOM/DSS, which induces colorectal cancer model, passes through carcinogen
(AOM) and to scorching agent (DSS) process of mankind's colorectal carcinoma occurrence and development is simulated.AOM is injected intraperitoneally, mouse intestines can be promoted
DNA methylation occurs for road normal cell, largely increases cell and the several of DNA mistakes pairing occur in breeding
Rate, subsequent three periods 2.5%DSS drinking-water feeding, destroy mouse Colon and rectum intestinal mucosa cells of superficial layer, in addition enteron aisle there are
A large amount of floras result in Colon and rectum inflammation, further promote the generation and growth of tumour.
During model foundation, the feeding of 2.5%DSS drinking-water can cause mouse apparent pus and blood stool occur, and with
Compared with batch normal rat of raising, weight is declined, it was demonstrated that the drinking-water of 2.5%DSS can lead to the hair of mouse Colon and rectum inflammation
It is raw.After 80 days model foundations, we have chosen department pattern mouse and control group mouse is put to death, dissected, observation knot
Rectum position situation.As shown in Figure 8, model mouse is the same as compared with batch normal rat, rectal intestinal tract length is obviously shortened, and can
See that the polypoid object that swells of prominent enteric cavity is formed.Then, we swell polypoid object and the intestines group with batch normal rat corresponding position
Taking-up is knitted, is placed in 4% paraformaldehyde fixed, progress paraffin section, passes through H＆E and dyes progress fabric analysis.Such as 9 institute of attached drawing
Show, visible glandular epithelium size, form are normal under control group mouse intestinal tissue mirror, have no heterocyst, have no tumour shape
Into.And found in model mouse intestinal tissue, normal gland structure disappears, and cell size, form differ, cell polar to disappearance,
And there are a large amount of inflammatory cell infiltrations, hence it is evident that visual tumors tissue.
AOM/DSS induces colorectal cancer mouse SPECT/CT imagings
It is as follows that AOM/DSS induces colorectal cancer model SPECT/CT imaging process flows：
First, carry out no Avidin pre-targeting imaging.Induction model mouse is taken, in tail vein injection 37MBq99mTc-
HYNIC-PEG3After-B, 2h, SPECT/CT nuclear medicines are carried out.
Second day carries out Avidin pre-targeting imagings.First is taken with only induction model mouse and with the normal of batch raising
Mouse is injected intraperitoneally 200 μ g Avidin and carries out pre-targeting, after 4h, tail vein injection 37MBq respectively99mTc-HYNIC-PEG3-
SPECT/CT nuclear medicines are carried out after B, 2h.After living imaging, mouse is put to death, dissect and is taken pictures, by whole enteron aisles
Tissue takes out, and is cleaned up intestinal contents using physiological saline, then carries out enteron aisle SPECT/CT nuclear medicines.Imaging
After, tumor tissues are taken out, is placed in 4% paraformaldehyde solution and fixes, for paraffin section, H＆E staining analysis.
We establish AOM/DSS and induce colorectal cancer tumor model, probe into Avidin/99mTc-HYNIC-PEG3- B is pre-
Targeted system is to the image checking ability of Colon and rectum primary tumo(u)r.As a result as shown in Figure 10, first day, no Avidin pre-targeting
SPECT/CT imaging experiments the result shows that (attached drawing 10c),99mTc-HYNIC-PEG3- B injects through tail vein and induces model mouse body
After interior, apparent radiated signal, abdominal cavity and other "dead" signals of normal structure organ are only detected at bladder.Second
Day, same induced model mouse and imaging experiment (attached drawing 10b) is carried out after Avidin pre-targeting, with acquired results phase on the firstth
Than in addition to bladder, it is dense poly- that apparent radiated signal at two occurs in abdominal cavity middle and lower part.After imaging, this model mouse is dissected, is seen
There is apparent bump (attached drawing 11-a, b) at two at rectum.Whole enteron aisles are taken out, and using PBS that enteron aisle content is clear
It removes, then carries out enteron aisle SPECT/CT and be imaged in vitro, it is seen that the cohesion of bump radioactivity is apparent, and normal bowel tissue is without radiation
Property signal (attached drawing 11-c).Finally, the intestinal tissue of protuberance is taken out, is placed in 4% formalin solution and fixes, carried out paraffin and cut
Piece, H＆E dyeing, determine to be tumor tissues (attached drawing 11-d, e) at two.
Then, in order to further verify Avidin/99mTc-HYNIC-PEG3- B is to the specificity of tumor imaging, Wo Men
With the SPECT/CT imaging experiments that Avidin pre-targeting has been carried out in batch normal rat body of raising, as a result such as attached drawing 10-a.99mTc-HYNIC-PEG3- B is absorbed in the intestinal tissue and histoorgan of normal mouse without apparent, is existed only in bladder.
This experimental result explanation：After Avidin is to inducing model mouse progress pre-targeting,99mTc-HYNIC-PEG3- B can
With the tumor locus concentrated at Colon and rectum of specificity, there is very strong in situ tumor imaging capability, and property is superior in vivo,
With higher tumour/non-tumour ratio.
SPECT/CT and archebiosis the light imaging of embodiment 6LS180-Luci colorectal cancer hepatic metastases
LS180-Luci colorectal cancer liver metastasis models are established
Splenic capsule injection-spleen reservation method, method detailed reference are taken in the foundation of LS180-Luci colorectal cancer liver metastasis models
Pertinent literature[3].Process is simply described as follows：4-5 week old BALB/c (nu/nu) nude mice abdominal cavity is taken to inject 1% yellow Jackets
75mg/kg is anaesthetized, and upper abdomen right-hand cutout is taken to enter abdomen after anesthesia, into abdomen after along greater curvature side expose spleen, ophthalmology Smooth forceps
Spleen is drawn out to outside notch by auxiliary medical cotton stick, and 29G insulin injection syringes paste splenic capsule inserting needle about 5mm at Jin Pi subordinates,
Slowly injection 0.05mL (5 × 106) under LS180-Luci colorectal cancer tumor cell suspensions to splenic capsule, it is seen that spleen at injection
Lighter and swelling oppress injection point 2min with the medical cotton stick that 75% alcohol infiltrates after injection, stop blooding and kill spilling
Spleen after confirming no bleeding, is also included in abdominal cavity, alcohol swab stick disinfection notch surrounding skin simultaneously closes abdomen, by model by tumour cell
Mouse is placed in SPF grades of environment raisings.After model foundation 2 weeks, after 100 μ L Luciferin of intraperitoneal injection (3mg/100 μ L), 10min,
Archebiosis light imaging is carried out using toy IVIS optical imaging systems, determines liver region metastases situation.
SPECT/CT and archebiosis the light imaging (BLI) of LS180-Luci colorectal cancer liver metastasis models
We are injected by splenic capsule-and spleen reservation method establishes LS180-Luci colorectal cancer liver metastasis models, and should
Archebiosis light (BLI) imaging is carried out with IVIS petty actions object optical system and determines tumour growth situation, and it is suitable big to treat that tumour reaches
It is small, carry out Avidin/99mTc-HYNIC-PEG3- B SPECT/CT are tested.As shown in result attached drawing 12, mouse whole body removes bladder
It is dense poly- more outer to locate radioactivity, left side liver region also shows apparent radiated signal aggregation, subsequent BLI it is experimentally confirmed that in
Nude mice liver lobus sinister lower edge is corresponding with SPECT/CT acquired results there are apparent colorectal cancer transfer stove.This experiment is demonstrate,proved
Real, Avidin can significantly be stranded in the metastatic tumour affected area of liver, and specificity intake after internal 4h is injected into99mTc-
HYNIC-PEG3- B carries out the metastatic lesion clearly SPECT/CT and is imaged, while have very high tumour/non-tumour ratio,
Result above proves Avidin/99mTc-HYNIC-PEG3B system can be used in the diagnosis of colorectal cancer hepatic metastases lesion.
The intermediate image of 1 attached drawing 10 of table and tumour, kidney probe intake degree (%ID) and tumour/kidney ratio in attached drawing 12
Value
| Induced tumor model (n=4) | Liver m etastases model (n=4) | |
| Tumor uptake (%ID) | 4.41±0.50 | 5.73±1.17 |
| Kidney absorbs (%ID) | 1.99±0.69 | 2.21±0.63 |
| Tumour/kidney | 2.43±0.92 | 2.91±1.44 |
Embodiment 799mTc-HYNIC-PEG3The medicine box of-B
Step 1 accurately weighs the HYNIC-PEG that embodiment 1 is prepared3-B 20μg。
Step 2 accurately weighs N- tri- (methylol) methylglycine (Tricine) 10mg.
Step 3 accurately weighs three sulfonate sodium (TPPTS) 5mg of triphenylphosphine.
Step 4 accurately weighs succinic acid 29.55mg.
The accurate extract water 1mL of step 5.
Step 6 accurately weighs sodium hydroxide 17mg.
More than substance is placed in 10mL cillin bottles by step 7, and concussion dissolving, filtering are placed in freeze drier and are lyophilized,
Obtain product medicine box.
Step 899mTc-HYNIC-PEG3- B preparation process：(1) medicine box 1 prepared is taken；(2) it is drawn with syringe
The Na of 1mL10-30mCi99mTcO4It is injected into medicine box, shakes mixing；(3) medicine box is placed in 100 DEG C of heating 20- in water-bath
30min.(4) it treats after reaction to take out medicine box, is placed in room temperature cooling, then sampling is analyzed using Radio-HPLC, analysis
Step, method are consistent with the hand labeled of embodiment 1, with HYNIC-PEG3- B synthesis HPLC analysis methods are identical.Prepare patent medicine
Mark rate is identical with 1 result of embodiment after box.
Bibliography：
[1]Tanaka T,Kohno H,Suzuki R et al.A novel inflammation-related mouse
colon carcinogenesis model induced by azoxymethane and dextran sodium sμ
Lfate.Cancer Sci.2003,94(11):965-973
[2]Neufert C,Becker C and Neurath MF.An inducible mouse model of
colon carcinogenesis for the analysis of sporadic and inflammation-driven
tumor progression.Nat Protoc.2007,2(8):1998-2004
[3]Price JE.Spontaneous and experimental metastasis models:nude
mice.Methods Mol Biol.2014,1070:223-233
More than, embodiments of the present invention are illustrated.But the present invention is not limited to the above embodiments.It is all
Within the spirit and principles in the present invention, any modification, equivalent substitution, improvement and etc. done should be included in the guarantor of the present invention
Within the scope of shield.
Claims (10)
- It is 1. a kind of99mThe radiolabeled complexes of Tc, the complex have a core sequestration ligand, the ligand with following formula L1 or Its stereoisomer form is its ligand precursor：Wherein：- L- is selected from linking arm as follows：The C that R is independently selected from H, is optionally optionally substituted by a hydroxyl group1-6Alkyl, C1-6Alkoxy, halogen ,-NH2；Integers of the n for 2-8, such as 2,3,4,5,6,7,8；Integers of the s for 0-6, such as 0,1,2,3,4,5,6；R1For H ,-SO3H ,-COOH or described sulfonic acid, carboxylic acid pharmaceutically acceptable salt or ester.
- 2. label complex according to claim 1, which is characterized in that the complex has the synergy modes of auxiliary, It is selected from water-soluble phosphine (such as three sulfonate sodium TPPTS of triphenylphosphine), N- tri- (methylol) methylglycine (Tricine), N- bis- (ethoxy) glycine, gluceptate, ethylenediamine-N, N '-diacetate esters (EDDA), 3- benzoyls Pyridine (BP), pyridine -2- azo-p- dimethylanilines (PADA)；Preferably, L is selected from the linking arm of formula d structures, and n is the integer of 3-5, and s is the integer of 2-3, it is preferable that n 3, s 2；Preferably, R1For H ,-SO3H ,-COOH；Preferably, synergy modes may be the same or different, independently selected from：N- tri- (methylol) methylglycine, triphenylphosphine three Between sulfonate sodium；Preferably, L1 is the compound or its stereoisomer of following formula：
- 3. complex is marked according to claim 1-2 any one of them, it is characterised in that the label complex is following formula：
- 4. a kind of molecular probe, it is characterised in that：It has such as claim 1-3 label complexs as defined in any one.
- 5. a kind of pharmaceutical composition, the composition includes a effective amount of claim 1-3 label cooperations as defined in any one Object.Preferably, described pharmaceutical composition is injectable formulation, the carrier containing the label complex and injectable.
- 6. pharmaceutical composition according to claim 5, which is characterized in that described pharmaceutical composition is a kind of radiodiagnosis Developer.It is preferred that the developer refers to the developer as SPECT.
- 7. a kind of drug system, the system includes the label complex and Avidin of any one of a effective amount of claim 1-3. Preferably, the drug system is a kind of kit, and complex and Avidin is marked to be divided in the kit.
- 8. a kind of any one of convenient preparation claim 1-399mTc marks the medicine box of complex, and the medicine box contains ligand precursor L1, synergy modes mixture lyophilized preparation.Preferably, the medicine box can be by that will contain ligand precursor L1, and cooperate with and match The buffer solution of body is lyophilized to be prepared into cillin bottle.When the preparation of complex is marked every time, the medicine prepared is taken out Box adds in corresponding exit dose99mTc sources, such as Na99mTcO4, at 80-120 DEG C, preferably 100 DEG C of water-baths, 10-60 points of heating Clock, preferably 20-30 minute, you can be made described99mTc marks complex.It is highly preferred that weigh the ligand precursor of the formula L1 15-25 μ g, such as 20 μ g；Weigh N- tri- (methylol) methylglycine (Tricine) 7.5-12.5mg, such as 10mg；It weighs Three sulfonate sodium (TPPTS) 3.75-6.25mg of triphenylphosphine, such as 5mg；It is a certain amount of to weigh succinic acid；Extract water is a certain amount of； More than substance is placed in cillin bottle and adjusts pH value to 5.5 using sodium hydroxide, is lyophilized after filtering and obtains medicine box.Particularly preferably Ground weighs the 20 μ g of Formula II ligand precursor described in claim 2, weighs N- tri- (methylol) methylglycine (Tricine) 10mg weighs three sulfonate sodium (TPPTS) 5mg of triphenylphosphine, weighs succinic acid 29.55mg, extract water 1mL weighs hydrogen-oxygen Change sodium 17mg, more than substance is placed in 10mL cillin bottles, be lyophilized after concussion dissolving, filtering, obtain product medicine box.
- 9. the application of the label complex of any one of claim 1-3 in medicine preparation, the drug is as radiodiagnosis Developer.
- 10. application according to claim 9, the drug is used to diagnose Malignancy and entity tumor, example Such as blood, liver, intestines, kidney, stomach, spleen, lung, muscle, bone position malignant tumour.It is preferred that those agglutinins Lectin receptors The cancer of height expression, preferably gastrointestinal cancer, such as colorectal cancer or the hepatic metastases by its initiation.
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| CN111991570A (en) * | 2020-07-24 | 2020-11-27 | 北京大学 | FAP-alpha specific tumor diagnosis SPECT imaging agent |
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