CN103570793A - Bile acid type compound, intermediate, preparation method and application of bile acid type compound - Google Patents
Bile acid type compound, intermediate, preparation method and application of bile acid type compound Download PDFInfo
- Publication number
- CN103570793A CN103570793A CN201310365746.8A CN201310365746A CN103570793A CN 103570793 A CN103570793 A CN 103570793A CN 201310365746 A CN201310365746 A CN 201310365746A CN 103570793 A CN103570793 A CN 103570793A
- Authority
- CN
- China
- Prior art keywords
- work
- compd
- time
- compound
- reaction
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- TVJSPSVDMUKDSN-SZQRXGOMSA-N C[C@H](CCC(NCCSNCC#C)=O)[C@@H](CC1)[C@@](C)(CC[C@@H]2[C@@](C)(CC[C@H](C3)OC(C)=O)[C@]3(C)C3)[C@@H]1[C@@H]2[C@@H]3OC(C)=O Chemical compound C[C@H](CCC(NCCSNCC#C)=O)[C@@H](CC1)[C@@](C)(CC[C@@H]2[C@@](C)(CC[C@H](C3)OC(C)=O)[C@]3(C)C3)[C@@H]1[C@@H]2[C@@H]3OC(C)=O TVJSPSVDMUKDSN-SZQRXGOMSA-N 0.000 description 1
Images
Landscapes
- Steroid Compounds (AREA)
Abstract
The invention discloses a bile acid type compound, an intermediate, a preparation method and an application of the bile acid type compound. The invention provides a bile acid type compound as shown in a formula A. When the 18F-labeled bile acid type compound A is applied as a positron emission tomography imaging agent, the great in-vivo and in-vitro stability is realized; when PET (positron emission tomography) imaging is performed, defluorination imaging is invisible; and furthermore, the bile acid type compound has the advantages of relatively fast in-vivo clearance speed and huge application potentials in research of liver function imaging, physiology and pathology, as well as diagnosis of diseases of a digestive system, in particular to the diagnosis of liver cancer and colon cancer.
Description
Technical field
The present invention relates to radiate video picture field, relate in particular to a kind of bile acids compound, intermediate and its preparation method and application.
Background technology
Primary hepatocarcinoma (abbreviation liver cancer) occupies the 5th in global cancer morbidity, and mortality ratio occupies the 3rd.In China; liver cancer annual death rate accounts for second (the Siegel AB of tumor mortality rate; Cohen EI; Ocean A, et al.Phase II Trial Evaluating the Clinical and Biologic Effects of Bevacizumab in Unresectable Hepatocellular Carcinoma.J Clin Oncol.2008; 26:2992-2998).The histological type of liver cancer have hepatocellular carcinoma (accounting for 91.5%), cholangiocellular carcinoma and Combination liver cancer (Li Linfa, Zhang Minming etc. tumor targeted molecular image. Beijing: Science Press, 2006,246-247).Liver cancer onset is relatively hidden, and lacks in early days special clinical manifestation, and most of patients has reached local late period or distant metastasis occurs when making a definite diagnosis, can not excision.Even liver cancer that can excision, 2 years recurrence rates are old ten thousand clear up to 50%(, Zou peasant, Zhang Siwei. Chinese mortality of liver cancer is deposited analysis [J] for geographical minute. and practical oncology magazine, 2008,3 (22): 201-203).Therefore, find early and diagnosing liver cancer to patient's treatment with prevent significant.
Positron emission robot calculator tomography (PET) has highly sensitive, the feature (Li Linfa such as resolving power is high, high specificity, Zhang Minming etc. tumor targeted molecular image. Beijing: Science Press, 2006,204), the early diagnosis and the post-operative evaluation that have been widely used at present tumour, have great researching value and market demand.Fluoro-18[
18f] there is good nulcear properties and chemical property, be the preferred nucleic of PET probe, be widely used in the mark of all kinds of PET molecules.
At present, the most frequently used PET molecular probe be fluorodeoxyglucose ([
18f] FDG), still, for the diagnosis of liver cancer, its sensitivity is lower.For the hepatocellular carcinoma PET video picture of having broken up without
18f-FDG gathers, PET video picture is negative or SUV not high, occur false negative result (Li Linfa, Zhang Minming etc. tumor targeted molecular image. Beijing: Science Press, 2006,246-247).Therefore, find the PET molecular imaging agent in each stage that a species specificity is higher, energy exact evaluation liver cancer develops, will not only be conducive to the early stage and staging diagnosis of liver cancer, and also significant for the evaluation of curative effect.
Bile acide is a kind of crude substance in human body, can be by liver receptor specific recognition, thereby become first-selected target molecule in the diagnosis of liver and gall diseases and treatment.Some investigators' discoveries, still can be identified by cholic acid haulage system the cholic acid after structure of modification, and especially 3 and 24 at cholic acid carry out structural modification (Kagedahl, M.; Swaan, P.W.; Redemann, C.T.Pharm.Res.1997,14,176.Stephen, Z.F.; Yurrrachek, E.C.; Sharit, R.Biochem.Pharmacol.1992,43,1969).Therefore, the compound of bile acids is carried out to mark
18f radio-labeling, finds a route that is applicable to synthetic and mark, and the novel liver cancer PET Imaging probe that maybe can develop a kind of excellent performance is that those skilled in the art wish the technical problem solving always.Because bile acide can experience liver sausage circulation, aspect liver function video picture, also having larger application potential.Meanwhile, because some bile acide (as taurolithocholic acid, lithocholic acid etc.) is the membrane receptor of G protein coupling receptor (TGR5) physiological level, TGR5 overexpression in colon cancer cell is found in research.Therefore, such PET Imaging probe also has certain value in the diagnosis and prognosis evaluation of colon cancer-related diseases.And above relevant research all rarely has report both at home and abroad, make this invention there is good novelty and better application value.
Summary of the invention
Technical problem to be solved by this invention is in order to overcome existing
18f radio-labeled compound is unsuitable for the low defect of diagnosis, sensitivity of liver cancer cell, provides a kind of bile acids compound, particularly
18f labeling bile acid compounds, intermediate and its preparation method and application.
The present invention solves the problems of the technologies described above by the following technical programs:
The invention provides a kind of suc as formula the bile acids compound shown in A,
Wherein, R
1, R
2and R
3be H or OH independently of one another; R
4for methylene radical,
or
described
in the carbon that is connected with carbonyl with in described A-NH is connected, described
in the ethylidene that is connected with sulfuryl with in described A-NH is connected; R
5for C
1～C
10straight-chain alkyl-sub-or C
1～C
10branched alkylidene, C
5～C
7cycloalkylidene, poly-sub-ethylene glycol, C
6～C
10arylidene, C
6～C
10arylidene-C
1～C
4alkylidene group or C
1～C
6alkylidene group-C
6～C
10arylidene-C
1～C
4alkylidene group, described C
6～C
10arylidene-C
1～C
4in alkylidene group-C
1～C
4alkylidene group is connected with the fluorine in described A;
Described poly-sub-ethylene glycol is following group:
N2=2～7 wherein.
In the present invention, described F is preferably
18f.When F is
18during F, described bile acids compound is
18f labeling bile acid compounds.When F is common fluorine element, described bile acids compound is reference compound A '.
In the present invention, described C
1～C
10straight-chain alkyl-sub-be preferably following group:
Described C
1～C
10branched alkylidene be preferably following group:
N1=0～8 wherein, preferably 1,2 or 3.
In the present invention, n2 preferably 2 or 3.
In the present invention
,described C
6～C
10arylidene is preferably
In the present invention, described C
6～C
10arylidene-C
1～C
4alkylidene group is preferably following group:
In the present invention, described C
1～C
6alkylidene group-C
6～C
10arylidene-C
1～C
4alkylidene group is preferably following arbitrary group:
n3=1～5，n4=0～4。
In the present invention, described C
5～C
7cycloalkylidene is preferably following group:
N5=1～3 wherein.
Compd A of the present invention is preferably,
A1: work as R
1=-H, R
2=-α OH, R
3=-α OH,
time compd A;
Or, A2: work as R
1=-H, R
2=-H, R
3=-α OH,
time compd A;
Or, A3: work as R
1=-H, R
2=-α OH, R
3=-H,
time compd A;
Or, A4: work as R
1=-H, R
2=-β OH, R
3=-H,
time compd A;
Or, A5: work as R
1=-α OH, R
2=-H, R
3=-H,
time compd A;
Or, A6: work as R
1=-H, R
2=-H, R
3=-H,
time compd A;
Or, A7: work as R
1=-H, R
2=-α OH, R
3=-α OH,
time compd A;
Or, A8: work as R
1=-H, R
2=-H, R
3=-α OH,
time compd A;
Or, A9: work as R
1=-H, R
2=-α OH, R
3=-α OH,
time compd A;
Or, A10: work as R
1=-H, R
2=-α OH, R
3=-α OH,
time compd A;
Or, A11: work as R
1=-H, R
2=-H, R
3=-α OH,
time compd A;
Or, A12: work as R
1=-H, R
2=-α OH, R
3=-H,
time compd A.
In the present invention ,-Z all refers to Z group, for example-CH
3nail base ,-OH refers to hydroxyl.
The present invention also provides the preparation method of bile acids compd A, and it comprises the steps: in solvent, under the catalysis of Cu (I), compd B and Compound D is carried out to nitrine and the Huisgen1 that holds position alkynyl, 3-Dipolar Cycloaddition;
Wherein, R
1, R
2, R
3, R
4and R
5definition ditto described in.
Wherein, described F is preferably
18f.
In the present invention, described Huisgen1, the method for 3-Dipolar Cycloaddition and condition can be method and condition used in this type of reaction of organic synthesis field, and described Cu (I) is monovalence copper, and generally the form with cupprous salt participates in reaction.
The inventor, through great many of experiments, particularly preferably goes out following method and condition: in solvent, pH is 3～12, under the catalysis of Cu (I), compd B and Compound D is carried out to nitrine and 1 of end position alkynyl, 3-Dipolar Cycloaddition.
Wherein, described pH is preferably 5～8.Described preferred solvents ground is water, the trimethyl carbinol, acetonitrile, tetrahydrofuran (THF), DMF(N, dinethylformamide) and DMSO(dimethyl sulfoxide (DMSO)) in one or more, the concentration of compd B in reaction solution is preferably 0.1～50mmol/L, and that better is 1～7mmol/L.Compound D is (2.0 * 10 with the molecular volume of solvent than preferably
-13mol～2.0 * 10
-7mol)/(0.2～1mL), the radioactive activity of Compound D in solvent is preferably 0.1mCi～4Ci.The amount of described Cu (I) is preferably 0.5 times～11 times of compd B molar weight, and better is 1.5 times～6 times.The concentration of described Cu (I) in reaction solution is preferably 1mol/L～5mol/L.
Described 1, the temperature of 3-Dipolar Cycloaddition can be according to the stability that participates in the compd B of reaction, and the boiling point of the reaction solvent system adopting, and suitably regulates temperature, is preferably 20～110 ℃, and better is 35～70 ℃.
Described 1,3-Dipolar Cycloaddition can complete in a short period of time, and as 1～80 minute, better was 10～20 minutes.
Wherein, described pH value can regulate by this area ordinary method, as adds the phosphate buffered saline buffer of required pH scope.
Described Cu (I) can be the common form of the Cu (I) of this type of reaction in organic field, the present invention particularly preferably Cu of following form (I) participates in reaction: cupric strong acid salt and xitix or its highly basic salt are carried out to reduction reaction, make Cu (I);
Described cupric strong acid salt can be one or more in copper sulfate, cupric nitrate and cupric chloride, preferably sulfuric acid copper.The highly basic salt of described xitix can be one or more in sodium ascorbate, potassium ascorbate and calcium ascorbate etc., because sodium ascorbate is more soluble in water than xitix, more common and cost is lower than other ascorbate salts, preferably sodium ascorbate.The mol ratio of described cupric strong acid salt and xitix or its highly basic salt is preferably 1:1.2～1:8, and that better is 1:2～1:4.
For described
18f labeling bile acid compounds, above-mentioned 1, after 3-Dipolar Cycloaddition finishes, available radioactivity HPLC separation and purification marked product, before radioactivity HPLC separation and purification, also can first carry out purifying with Sep-Pak C18 post to product.
The solution decompression that HPLC separation and purification is obtained is concentrated, then is dissolved in the mixing solutions (ethanol content <10%) of ethanol and physiological saline, through aseptic membrane filtration, obtains
18f mark preparation.
Described
18the vitro stability of F labeling bile acid compounds is measured
At PBS(phosphate buffer soln) in stability: get described
18 f mark preparation 50 μ L(approximately 50 μ Ci), be placed in 1mL PBS, be placed at 37 ℃, after oscillation incubation 30,60,120,180,240min, with HPLC, measure its radiochemical purity, to observe its stability in PBS.
Stability in serum: described in getting
18 f mark preparation 50 μ L(approximately 50 μ Ci), be placed in 1000 μ L calf serums, be placed at 37 ℃, after oscillation incubation 30,60,120,240,360min, sample 100 μ L and be placed in the dactylethrae that contains 200 μ L methyl alcohol, mix latter centrifugal two minutes, get supernatant liquor and measure its radiochemical purity with HPLC, to observe its stability in serum.
Described
18the mensuration of the lipotropy of F labeling bile acid compounds (Log P)
Log P is the lipid of compound, can reflect a lipophilic power of compound.It measures calculation formula:
Get 10 μ L
18f mark preparation (contains 600 μ L n-Octanols and 590 μ LPBS) in 2mL dactylethrae, good seal, and at room temperature, after abundant vortex 2min, high speed centrifugation 3min, to biphase equilibrium.With pipettor, from organic phase and water, respectively sample 100 μ L and be placed in two V counter tubes, with gamma counter, measure counting.Repeated sampling is measured three times.Parallelly carry out two batches of experiments, calculate
18the average Log P value of F mark preparation.
In the present invention, described Compound D can be made by following method: by compd E and
18f
-carry out nucleophilic substitution reaction:
Wherein, R
6for leavings group conventional in nucleophilic substitution reaction, as-OTs ,-OMs or-OTf, R
5definition ditto described in.
Wherein, described F is
18during F, the method for described nucleophilic substitution reaction and condition can be this area this type of
18the ordinary method of F labeled reactant and condition, the present invention is following method and condition particularly preferably: in organic solvent, under protection of inert gas, will contain K
222, K
2cO
3with
18f
-mixture and compd E carry out nucleophilic substitution reaction.
Wherein, described organic solvent is preferably one or more in anhydrous acetonitrile, anhydrous dimethyl formamide and anhydrous dimethyl sulfoxide, preferably anhydrous acetonitrile.Described K
222and K
2cO
3mol ratio be preferably 1:4～8:1, that better is 1:1～3:1.
18f
-activity be preferably 10 μ Ci～5Ci, that better is 5mCi～1Ci.The concentration of compd E in reaction solution is preferably 0.005～1mol/L, and that better is 0.05～0.25mol/L.K
222with the mass ratio of compd E be preferably 1:2～8:1, that better is 2:1-4.5:1.Described rare gas element is preferably nitrogen and/or argon gas.The temperature of described nucleophilic substitution reaction is preferably 80～150 ℃.The time of described nucleophilic substitution reaction is preferably 2～15min.
The described K that contains
222, K
2cO
3with
18f
-mixture can make by following method: use K
222(being Kryptofix222) solution drip washing enrichment
18f
-qMA post, solvent evaporated.
Wherein, K
222solution can make by following method: by K
222, K
2cO
3, acetonitrile and water wiring solution-forming.Wherein, each component content scope is as follows: in every 1mL acetonitrile, have 30～140 μ L water, 1～8mg K
2cO
3, 4.5～25mg K
222.The method of configuration can be for adding 30～140 μ L water, 1～8mg K in 1mL acetonitrile
2cO
3, 4.5～25mg K
222.The most frequently used a kind of proportioning is: in every 960 μ L acetonitriles, have 14.4mg K
222, 3.0mgK
2cO
3, 40 μ L water, wiring solution-forming.
After above-mentioned nucleophilic substitution reaction completes, aftertreatment and the method for purification of available this area routine are purified.The preferred following method of purification of the present invention and condition: when the boiling point of Compound D is during lower than 200 ℃, in reaction solution, add acetonitrile, using nitrogen as carrier gas, adopt distillating method separating impurity, collect the acetonitrile condensing soln of Compound D.Described distillation temperature is preferably 85～150 ℃, and distillation time is preferably 5～30 minutes.
In the present invention, described compd B can be made by following method:
When
time, in organic solvent, Compound C x is reacted with propargylamine;
When
time, in organic solvent, Compound C y is reacted with propargylamine;
Wherein, R
1, R
2and R
3definition ditto described in.
Described compd B is preferably,
B1: work as R
1=-H, R
2=-α OH, R
3=-α OH,
time compd B;
Or, B2: work as R
1=-H, R
2=-H, R
3=-α OH,
time compd B;
Or, B3: work as R
1=-H, R
2=-α OH, R
3=-H,
time compd B;
Or, B4: work as R
1=-H, R
2=-β OH, R
3=-H,
time compd B;
Or, B5: work as R
1=-α OH, R
2=-H, R
3=-H,
time compd B;
Or, B6: work as R
1=-H, R
2=-H, R
3=-H,
time compd B;
Or, B7: work as R
1=-H, R
2=-α OH, R
3=-α OH,
time compd B;
Or, B8: work as R
1=-H, R
2=-H, R
3=-α OH,
time compd B;
Or, B9: work as R
1=-H, R
2=-α OH, R
3=-α OH,
time compd B;
Described Compound C x is preferably,
C1: work as R
1=-H, R
2=-α OH, R
3compound C x during=-α OH;
Or, C2: work as R
1=-H, R
2=-H, R
3compound C x during=-α OH;
Or, C3: work as R
1=-H, R
2=-α OH, R
3compound C x during=-H;
Or, C4: work as R
1=-H, R
2=-β OH, R
3compound C x during=-H;
Or, C5: work as R
1=-α OH, R
2=-H, R
3compound C x during=-H;
Or, C6: work as R
1=-H, R
2=-H, R
3compound C x during=-H;
Described Compound C y is preferably,
C7: work as R
1=-H, R
2=-α OH, R
3compound C y during=-α OH;
Or, C8: work as R
1=-H, R
2=-H, R
3compound C y during=-α OH;
Or, C9: work as R
1=-H, R
2=-α OH, R
3compound C y during=-α OH.
Wherein, the method of described reaction and condition can be ordinary method and the condition of this type of reaction of this area, the present invention is following method and condition particularly preferably: in the mixture of methylene dichloride, 1-ethyl-3-(3-dimethylaminopropyl) phosphinylidyne diimmonium salt hydrochlorate (EDC.HCl), I-hydroxybenzotriazole (HOBt) and triethylamine, Compound C x or Cy are reacted with propargylamine.Wherein, the temperature of described reaction is preferably 10～30 ℃.Described Compound C x or Cy:1-ethyl-3-(3-dimethylaminopropyl) phosphinylidyne diimmonium salt hydrochlorate: I-hydroxybenzotriazole: triethylamine=1:(1～5): (1～5): (1～8).After this reaction completes, aftertreatment and the method for purification of available this area routine are purified.The preferred following method of purification of the present invention and condition: water, saturated sodium bicarbonate solution, dilute hydrochloric acid are washed once successively, then wash twice with sodium chloride solution, after organic layer anhydrous sodium sulfate drying, column chromatography purification (developping agent: sherwood oil, ethyl acetate).
Work as R
1=-H, R
2=-α OH, R
3=-α OH,
time, the preparation method of compd B can comprise the steps:
1. step, reacts Compound C 10 with diacetyl oxide, obtain C10-1;
2. step, reacts C10-1 with sulfur oxychloride, obtain C10-2;
3. step, reacts C10-2 with propargylamine, obtain C10-3;
4. step, reacts C10-3 with hydrochloric acid, obtain compd B;
Work as R
1=-H, R
2=-H, R
3=-α OH,
time, the preparation method of compd B can comprise the steps:
1. step, reacts Compound C 11 with diacetyl oxide, obtain C11-1;
2. step, reacts C11-1 with sulfur oxychloride, obtain C11-2;
3. step, reacts C11-2 with propargylamine, obtain C11-3;
4. step, reacts C11-3 with hydrochloric acid, obtain compd B;
Work as R
1=-H, R
2=-α OH, R
3=-H,
time, the preparation method of compd B can comprise the steps:
1. step, reacts Compound C 12 with diacetyl oxide, obtain C12-1;
2. step, reacts C12-1 with sulfur oxychloride, obtain C12-2;
3. step, reacts C12-2 with propargylamine, obtain C12-3;
4. step, reacts C12-3 with hydrochloric acid, obtain compd B;
Wherein, the method for described step reaction 1. and condition can be ordinary method and the condition of this type of reaction of this area, and the present invention is following method and condition particularly preferably: in pyridine, Compound C 10 is reacted with diacetyl oxide, obtain C10-1; Or, in pyridine, Compound C 11 is reacted with diacetyl oxide, obtain C11-1; Or, in pyridine, Compound C 12 is reacted with diacetyl oxide, obtain C12-1.The temperature of this reaction is preferably 25 ℃.For C10, the mol ratio of described C10 and described diacetyl oxide is preferably 1:3～1:20, is more preferably 1:4; For C11 or C12, the mol ratio of described C11 or C12 and described diacetyl oxide is preferably 1:2～1:10, is more preferably 1:3.After this reaction completes, aftertreatment and the method for purification of available this area routine are purified.The preferred following method of purification of the present invention and condition: in reaction solution, add distilled water, with 1mmol/L hydrochloric acid, be adjusted to without solid and separate out, filter, filter cake distilled water wash, grinds dry.
Wherein, the method of described step reaction 2. and condition can be ordinary method and the condition of this type of reaction of this area, the present invention is following method and condition particularly preferably: C10-1 and sulfur oxychloride are added in the 50mL round-bottomed flask that drying tube, prolong are housed, oil bath heating, 80 ℃ of stirring reaction 6h, remove solvent under reduced pressure in 40 ℃; Or, C11-1 and sulfur oxychloride are added in the 50mL round-bottomed flask that drying tube, prolong are housed, oil bath heating, 80 ℃ of stirring reaction 6h, remove solvent under reduced pressure in 40 ℃; Or, C12-1 and sulfur oxychloride are added in the 50mL round-bottomed flask that drying tube, prolong are housed, oil bath heating, 80 ℃ of stirring reaction 6h, remove solvent under reduced pressure in 40 ℃.
Wherein, the method for described step reaction 3. and condition can be ordinary method and the condition of this type of reaction of this area, and the present invention is following method and condition particularly preferably: in the mixture of DMF and pyridine, C10-2 is reacted with propargylamine; Or, in the mixture of DMF and pyridine, C11-2 is reacted with propargylamine; Or, in the mixture of DMF and pyridine, C12-2 is reacted with propargylamine.The mol ratio of described C10-2, C11-2 or C12-2 and described propargylamine is preferably 1:1～1:3.After this reaction completes, aftertreatment and the method for purification of available this area routine are purified.The preferred following method of purification of the present invention and condition: filter, remove solvent under reduced pressure, add distilled water to stir and separate out solid, filter, be dried, with purification by silica gel column chromatography, petrol ether/ethyl acetate gradient elution, collect elutriant, remove solvent under reduced pressure, solid drying.
Wherein, the method for described step reaction 4. and condition can be ordinary method and the condition of this type of reaction of this area, and the present invention is following method and condition particularly preferably: in DMF, C10-3 is reacted with hydrochloric acid, obtain compd B; Or, in DMF, C11-3 is reacted with hydrochloric acid, obtain compd B; Or, in DMF, C12-3 is reacted with hydrochloric acid, obtain compd B.The volumetric concentration of described hydrochloric acid is preferably 36.5%; Described C10-3, C11-3 or C12-3 are 0.1～0.5mol/ml with the molecular volume of described hydrochloric acid than preferably.After this reaction completes, aftertreatment and the method for purification of available this area routine are purified.The preferred following method of purification of the present invention and condition: remove solvent under reduced pressure, with purification by silica gel column chromatography, petrol ether/ethyl acetate gradient elution, collect elutriant, remove solvent under reduced pressure, solid drying.
The present invention also provides compd B,
Wherein, R
1, R
2and R
3be H or OH independently of one another, R
4for methylene radical,
or
Wherein, described compd B can be used as the intermediate for the preparation of above-mentioned bile acids compd A.
In the present invention, described compd B is preferably,
B1: work as R
1=-H, R
2=-α OH, R
3=-α OH,
time compd B;
Or, B2: work as R
1=-H, R
2=-H, R
3=-α OH,
time compd B;
Or, B3: work as R
1=-H, R
2=-α OH, R
3=-H,
time compd B;
Or, B4: work as R
1=-H, R
2=-β OH, R
3=-H,
time compd B;
Or, B5: work as R
1=-α OH, R
2=-H, R
3=-H,
time compd B;
Or, B6: work as R
1=-H, R
2=-H, R
3=-H,
time compd B;
Or, B7: work as R
1=-H, R
2=-α OH, R
3=-α OH,
time compd B;
Or, B8: work as R
1=-H, R
2=-H, R
3=-α OH,
time compd B;
Or, B9: work as R
1=-H, R
2=-α OH, R
3=-α OH,
time compd B;
Or, B10: work as R
1=-H, R
2=-α OH, R
3=-α OH,
time compd B;
Or, B11: work as R
1=-H, R
2=-H, R
3=-α OH,
time compd B;
Or, B12: work as R
1=-H, R
2=-α OH, R
3=-H,
time compd B.
The present invention also provides Compound C 10-1, C10-2, C10-3, C11-1, C11-2, C11-3, C12-1, C12-2 and C12-3,
Wherein, described Compound C 10-1, C10-2, C10-3, C11-1, C11-2, C11-3, C12-1, C12-2 and C12-3 can be used as the intermediate for the preparation of above-mentioned bile acids compd A.
The present invention also provides the preparation method of above-claimed cpd B, and it comprises the steps:
When
time, in organic solvent, Compound C x is reacted with propargylamine;
When
time, in organic solvent, Compound C y is reacted with propargylamine;
Wherein, R
1, R
2and R
3definition ditto described in.
Described compd B is preferably,
B1: work as R
1=-H, R
2=-α OH, R
3=-α OH,
time compd B;
Or, B2: work as R
1=-H, R
2=-H, R
3=-α OH,
time compd B;
Or, B3: work as R
1=-H, R
2=-α OH, R
3=-H,
time compd B;
Or, B4: work as R
1=-H, R
2=-β OH, R
3=-H,
time compd B;
Or, B5: work as R
1=-α OH, R
2=-H, R
3=-H,
time compd B;
Or, B6: work as R
1=-H, R
2=-H, R
3=-H,
time compd B;
Or, B7: work as R
1=-H, R
2=-α OH, R
3=-α OH,
time compd B;
Or, B8: work as R
1=-H, R
2=-H, R
3=-α OH,
time compd B;
Or, B9: work as R
1=-H, R
2=-α OH, R
3=-α OH,
time compd B;
Described Compound C x is preferably,
C1: work as R
1=-H, R
2=-α OH, R
3compound C x during=-α OH;
Or, C2: work as R
1=-H, R
2=-H, R
3compound C x during=-α OH;
Or, C3: work as R
1=-H, R
2=-α OH, R
3compound C x during=-H;
Or, C4: work as R
1=-H, R
2=-β OH, R
3compound C x during=-H;
Or, C5: work as R
1=-α OH, R
2=-H, R
3compound C x during=-H;
Or, C6: work as R
1=-H, R
2=-H, R
3compound C x during=-H;
Described Compound C y is preferably,
C7: work as R
1=-H, R
2=-α OH, R
3compound C y during=-α OH;
Or, C8: work as R
1=-H, R
2=-H, R
3compound C y during=-α OH;
Or, C9: work as R
1=-H, R
2=-α OH, R
3compound C y during=-α OH.
Wherein, the method of described reaction and condition can be ordinary method and the condition of this type of reaction of this area, the present invention is following method and condition particularly preferably: in the mixture of methylene dichloride, 1-ethyl-3-(3-dimethylaminopropyl) phosphinylidyne diimmonium salt hydrochlorate (EDC.HCl), I-hydroxybenzotriazole (HOBt) and triethylamine, Compound C x or Cy are reacted with propargylamine.Wherein, the temperature of described reaction is preferably 10～30 ℃.Described Compound C x or Cy:1-ethyl-3-(3-dimethylaminopropyl) phosphinylidyne diimmonium salt hydrochlorate: I-hydroxybenzotriazole: triethylamine=1:(1～5): (1～5): (1～8).After this reaction completes, aftertreatment and the method for purification of available this area routine are purified.The preferred following method of purification of the present invention and condition: water, saturated sodium bicarbonate solution, dilute hydrochloric acid are washed once successively, then wash twice with sodium chloride solution, after organic layer anhydrous sodium sulfate drying, column chromatography purification (developping agent: sherwood oil, ethyl acetate).
Work as R
1=-H, R
2=-α OH, R
3=-α OH,
time, the preparation method of compd B can comprise the steps:
1. step, reacts Compound C 10 with diacetyl oxide, obtain C10-1;
2. step, reacts C10-1 with sulfur oxychloride, obtain C10-2;
3. step, reacts C10-2 with propargylamine, obtain C10-3;
4. step, reacts C10-3 with hydrochloric acid, obtain compd B;
Work as R
1=-H, R
2=-H, R
3=-α OH,
time, the preparation method of compd B can comprise the steps:
1. step, reacts Compound C 11 with diacetyl oxide, obtain C11-1;
2. step, reacts C11-1 with sulfur oxychloride, obtain C11-2;
3. step, reacts C11-2 with propargylamine, obtain C11-3;
4. step, reacts C11-3 with hydrochloric acid, obtain compd B;
Work as R
1=-H, R
2=-α OH, R
3=-H,
time, the preparation method of compd B can comprise the steps:
1. step, reacts Compound C 12 with diacetyl oxide, obtain C12-1;
2. step, reacts C12-1 with sulfur oxychloride, obtain C12-2;
3. step, reacts C12-2 with propargylamine, obtain C12-3;
4. step, reacts C12-3 with hydrochloric acid, obtain compd B;
Wherein, the method for described step reaction 1. and condition can be ordinary method and the condition of this type of reaction of this area, and the present invention is following method and condition particularly preferably: in pyridine, Compound C 10 is reacted with diacetyl oxide, obtain C10-1; Or, in pyridine, Compound C 11 is reacted with diacetyl oxide, obtain C11-1; Or, in pyridine, Compound C 12 is reacted with diacetyl oxide, obtain C12-1.The temperature of this reaction is preferably 25 ℃.For C10, the mol ratio of described C10 and described diacetyl oxide is preferably 1:3～1:20, is more preferably 1:4; For C11 or C12, the mol ratio of described C11 or C12 and described diacetyl oxide is preferably 1:2～1:10, is more preferably 1:3.After this reaction completes, aftertreatment and the method for purification of available this area routine are purified.The preferred following method of purification of the present invention and condition: in reaction solution, add distilled water, with 1mmol/L hydrochloric acid, be adjusted to without solid and separate out, filter, filter cake distilled water wash, grinds dry.
Wherein, the method of described step reaction 2. and condition can be ordinary method and the condition of this type of reaction of this area, the present invention is following method and condition particularly preferably: C10-1 and sulfur oxychloride are added in the 50mL round-bottomed flask that drying tube, prolong are housed, oil bath heating, 80 ℃ of stirring reaction 6h, remove solvent under reduced pressure in 40 ℃; Or, C11-1 and sulfur oxychloride are added in the 50mL round-bottomed flask that drying tube, prolong are housed, oil bath heating, 80 ℃ of stirring reaction 6h, remove solvent under reduced pressure in 40 ℃; Or, C12-1 and sulfur oxychloride are added in the 50mL round-bottomed flask that drying tube, prolong are housed, oil bath heating, 80 ℃ of stirring reaction 6h, remove solvent under reduced pressure in 40 ℃.
Wherein, the method for described step reaction 3. and condition can be ordinary method and the condition of this type of reaction of this area, and the present invention is following method and condition particularly preferably: in the mixture of DMF and pyridine, C10-2 is reacted with propargylamine; Or, in the mixture of DMF and pyridine, C11-2 is reacted with propargylamine; Or, in the mixture of DMF and pyridine, C12-2 is reacted with propargylamine.The mol ratio of described C10-2, C11-2 or C12-2 and described propargylamine is preferably 1:1～1:3.After this reaction completes, aftertreatment and the method for purification of available this area routine are purified.The preferred following method of purification of the present invention and condition: filter, remove solvent under reduced pressure, add distilled water to stir and separate out solid, filter, be dried, with purification by silica gel column chromatography, petrol ether/ethyl acetate gradient elution, collect elutriant, remove solvent under reduced pressure, solid drying.
Wherein, the method for described step reaction 4. and condition can be ordinary method and the condition of this type of reaction of this area, and the present invention is following method and condition particularly preferably: in DMF, C10-3 is reacted with hydrochloric acid, obtain compd B; Or, in DMF, C11-3 is reacted with hydrochloric acid, obtain compd B; Or, in DMF, C12-3 is reacted with hydrochloric acid, obtain compd B.The volumetric concentration of described hydrochloric acid is preferably 36.5%; Described C10-3, C11-3 or C12-3 are 0.1～0.5mol/ml with the molecular volume of described hydrochloric acid than preferably.After this reaction completes, aftertreatment and the method for purification of available this area routine are purified.The preferred following method of purification of the present invention and condition: remove solvent under reduced pressure, with purification by silica gel column chromatography, petrol ether/ethyl acetate gradient elution, collect elutriant, remove solvent under reduced pressure, solid drying.
It is above-mentioned that the present invention also provides
18f labeling bile acid compounds A is as the application of positron emission tomography agent.
Without prejudice to the field on the basis of common sense, above-mentioned each optimum condition, can arbitrary combination, obtains the preferred embodiments of the invention.
Agents useful for same of the present invention and raw material be commercially available obtaining all.
Positive progressive effect of the present invention is:
1, simple for radiolabeled precursor synthesis step in the present invention, productive rate is high, is easy to a large amount of preparations.
2, PET developer preparation method of the present invention is simple, and radio chemistry productive rate is high, and the reaction times is short, is more applicable to radiopharmaceuticals clinical application, prepares
18f tagged compound radiochemical purity is greater than 99%.
3, PET developer of the present invention has in good body and vitro stability, has no defluorinate video picture during PET video picture.
4, the present invention
18the bile acids developer of F mark has faster removing speed in body, the diagnosis of liver function video picture, physiology and pathological research and digestive system particularly the diagnosis of liver cancer and colorectal carcinoma have huge application potential.
5, in the present invention, copper sulfate used, sodium ascorbate etc. are commercialization reagent, and raw material is cheap and easy to get.
Accompanying drawing explanation
Fig. 1 is that the marked product A6-second of example one of embodiment 4 37 ℃ of Radio-HPLC while hatching 360min in PBS detect spectrogram.
Fig. 2 is that the marked product A6-second of example one of embodiment 4 37 ℃ of Radio-HPLC while hatching 360min in calf serum detect spectrogram.
Fig. 3 is that the marked product A3-second of example two of embodiment 4 37 ℃ of Radio-HPLC while hatching 360min in calf serum detect spectrogram.
Fig. 4 is that the marked product A3-second of example two of embodiment 4 37 ℃ of Radio-HPLC while hatching 360min in PBS detect spectrogram.
Striograph when Fig. 5 is the PET-CT scanning 1min of example two marked product A3-second of embodiment 4.
Striograph when Fig. 6 is the PET-CT scanning 10min of example two marked product A3-second of embodiment 4.
Striograph when Fig. 7 is the PET-CT scanning 1h of example two marked product A3-second of embodiment 4.
Fig. 8 is that the preparation of example two marked product A3-second of embodiment 4 is at the SUV-t of the liver of normal mice curve.
Embodiment
Mode below by embodiment further illustrates the present invention, but does not therefore limit the present invention among described scope of embodiments.The experimental technique of unreceipted actual conditions in the following example, according to ordinary method and condition, or selects according to catalogue.
Embodiment 1
Synthesizing of labelled precursor compound
Synthesizing of [example one], CDCA acyl propargylamine (B3)
Chenodiol 393mg (1mmol) joins 30mL CH
2cl
2in, under stirring, add EDC.HCl230mg (1.2mmol), HOBt162mg (1.2mmol), triethylamine 280 μ L (2mmol), add propargylamine 82.5 μ L(1.2mmol after stirred for several minute).Above-mentioned reaction solution stirring at room reaction two days.Once, saturated sodium bicarbonate solution is washed once in reaction solution washing, and dilute hydrochloric acid is washed once, and saturated nacl aqueous solution washes twice, after organic layer anhydrous sodium sulfate drying, and column chromatography purification (developping agent: sherwood oil, ethyl acetate).Obtain white solid 315.6mg.Yield is 73.5%, purity 99%(HPLC).
The appraising datum of B3 is as follows: TOF-ESI-MS:M (C
27h
43nO
3)=429.64 (m/z), 452.3[M+Na]
+.
13C?NMR(400MHz,CDCl
3)δ:11.79，18.39，20.58，22.28，23.73，28.21，29.19，30.67，31.54，32.84，33.25，34.62，35.05，35.33，35.45，39.43，39.65，39.91，41.48，42.71，50.47，55.81，68.53，71.56，72.02，79.70，173.09.
1H-NMR(400MHz,CDCl
3)δ5.70(br?s,1H,NH),δ4.06(d,1H,C
25-αH,J=2.4),δ4.04(d,1H,C
25-βH,J=2.4),δ3.85(m,1H,C
7-H),δ3.46(m,C
3-H),δ2.3-0.92（m,32H）,δ0.90(s,3H,C
19-H),δ0.66(s,3H,C
18-H).
Synthesizing of [example two], stone courage acyl propargylamine (B6)
Lithocholic acid 377mg (1mmol) joins in 30mL CH2Cl2, under stirring, add EDC.HCl 230mg (1.2mmol), HOBt162mg (1.2mmol), triethylamine 280 μ L (2mmol), add propargylamine 82.5 μ L(1.2mmol after stirred for several minute).Above-mentioned reaction solution stirring at room reaction two days.Once, saturated sodium bicarbonate solution is washed once in reaction solution washing, and dilute hydrochloric acid is washed once, and saturated nacl aqueous solution washes twice, after organic layer anhydrous sodium sulfate drying, and column chromatography purification (developping agent: sherwood oil, ethyl acetate).Obtain white solid 307.5mg, yield is 74.3%, purity 99%(HPLC).
The appraising datum of B6 is as follows: TOF-ESI-MS:M (C
27h
43nO
2)=413.64 (m/z), 414.4[M+H]
+, 436.4[M+Na], 827.8[2M+H
+], 849.8[2M+Na
+].
13C?NMR(400MHz,CDCl
3)δ:12.06，18.39，20.83，23.38，24.21，26.42，27.20，28.26，29.19，30.56，31.57，33.31，34.58，35.35，35.45，35.86，36.47，40.19，40.44，42.10，42.76，55.98，56.51，71.56，71.89，79.69，173.11.
1H-NMR(400MHz,CDCl
3)δ5.64(br?s,1H,NH),δ4.06(d,1H,C
25-αH,J=2.4),δ4.04(d,1H,C
25-βH,J=2.4),δ3.63(m,1H,C
3-H),δ2.3-0.92（m,32H）,δ0.92(s,3H,C
19-H),δ0.64(s,3H,C
18-H).
Synthesizing of [example three], cow-bezoar CDCA acyl propargylamine (B12)
C12-1's is synthetic:
By C12750mg(1.5mmol), diacetyl oxide 142uL(1.5mmol) and pyridine 8mL be added in 50mL round-bottomed flask, under 25 ℃ of conditions, stirring reaction spends the night.In reaction solution, add 60mL distilled water, with 1mmol/L hydrochloric acid, be adjusted to without solid and separate out, filter, filter cake distilled water wash, grinds and is dried, and obtains product C 12-1.
C12-2's is synthetic:
C12-1 (1mmol) and sulfur oxychloride (5mL) are added in the 50mL round-bottomed flask that drying tube, prolong are housed, oil bath heating, 80 ℃ of stirring reaction 6h, remove solvent under reduced pressure in 40 ℃ and obtain compound (C12-2).
C12-3's is synthetic:
In the 100mL three-necked bottle of drying tube, constant pressure funnel is housed, add 2-propargyl amine 96uL(1.5mmol), dry DMF(5mL) and pyridine 121uL(1.5mmol), under ice bath, slowly drip the above-mentioned acyl chlorides (C12-2 for preparing, DMF solution 5mL 1mmol), about 0.5h drips complete, stirring reaction 20min, remove ice bath, stirring at room reaction is spent the night, and filters, and removes solvent under reduced pressure, add distilled water (20mL) to stir and separate out solid, filter, be dried.Crude product purification by silica gel column chromatography, petrol ether/ethyl acetate gradient elution, collects and merges product elutriant, removes solvent under reduced pressure, and solid drying obtains product C 12-3.
B12's is synthetic:
C12-3 (0.5mmol) is added to 50mL round-bottomed flask, add 5mL DMF and 1mL concentrated hydrochloric acid, 50 ℃ of stirring reaction 1h, are cooled to room temperature, remove solvent under reduced pressure, with purification by silica gel column chromatography, petrol ether/ethyl acetate gradient elution, collects and merges product elutriant, remove solvent under reduced pressure, solid drying obtains product B 12, yield 55%, purity 97%(HPLC).
Synthesizing of reference compound
Synthesizing of [example one], the corresponding reference compound A ' of A3-second 3-second
Under magnetic agitation, in 2mL pH=6.0 phosphate buffered saline buffer, successively add 170 μ L copper-baths (0.45mol/L) and 100 μ L sodium ascorbate solutions (1.5mol/L), after mixing, add B3 (25 μ mol, DMF solution 11mg) (500 μ L), acetonitrile solution (the 125 μ mol that add 2-nitrine-1-fluoroethane after 10min clock, V=3mL), 50 ℃ of stirring reaction 3h.Be cooled to room temperature, reaction solution adds the dilution of 30mL water, chloroform extraction three times (15mL/ time) for product.Merge organic layer, with saturated nacl aqueous solution, 15mL washes organic layer, and after concentrating under reduced pressure, obtaining product is white solid.Quality: 11.2mg, yield: 86.5%, purity 99%(HPLC).
The appraising datum of A ' 3-second is as follows: TOF-ESI-MS:M (C
29h
47fN
4o
3)=518.71 (m/z), 519.5[M+H]
+, 541.4[M+Na]
+.
1H-NMR(400MHz,CDCl
3)δ7.67(s,1H,CH),δ6.35(br?s,1H,NH),δ4.85(t,1H,-CHF,J=4.8),δ4.73(t,1H,-CHF,J=4.8),δ4.68(t,1H,CH,J=4.8),δ4.61(t,1H,CH,J=4.8)，δ4.51(d,2H,CH
2,J=3.6),δ3.85(d,1H,CH,J=2.4),δ3.46(m,1H,CH),δ2.96(s,1H,OH),δ2.88(s,1H,OH),δ2.3-0.9(m,29H),δ0.90(s,3H,C
19-H),δ0.64(s,3H,C
18-H)
13C?NMR(400MHz,CDCl
3)δ:173.70，129.74，123.25，82.24，80.52，72.01，68.51，55.71，50.67，42.70，41.50，39.89，39.64，39.45，35.48，35.33，35.05，34.81，34.60，33.30，32.84，31.59，30.68，28.21，23.71，22.78，20.58，18.38，11.76
Synthesizing of [example two], the corresponding reference compound A ' of A6-second 6-second
Under magnetic agitation, in 1.5mL pH=6.0 phosphate buffered saline buffer, successively add 130 μ L copper-baths (0.45mol/L) and 80 μ L sodium ascorbate solutions (1.5mol/L), after mixing, add B6 (20umol, DMF solution (400uL) 8.3mg), acetonitrile solution (the 100umol that adds 2-nitrine-1-fluoroethane after 10min clock, V=2.4mL), 50 ℃ of stirring reaction 3h.Be cooled to room temperature, reaction solution adds the dilution of 25mL water, chloroform extraction three times (15mL/ time) for product.Merge organic layer, with saturated nacl aqueous solution, 15mL washes organic layer, and after concentrating under reduced pressure, obtaining product is white solid.Quality: 8.6mg, yield: 85.3%, purity 99%(HPLC).
The appraising datum of A ' 6-second is as follows: TOF-ESI-MS:M (C
29h
47fN
4o
2)=502.71 (m/z), 503.55[M+H]
+, 525.55[M+Na]
+.
1H-NMR(400MHz,CDCl3)δ7.65(s,1H,CH),δ6.18(br?s,1H,NH),δ4.85(t,1H,-CHF,J=4.8),δ4.73(t,1H,-CHF,J=4.8),δ4.68(t,1H,CH,J=4.8),δ4.61(t,1H,CH,J=4.8),δ4.52(d,2H,CH
2,J=5.6),δ3.62(m,1H,C
3H),δ2.25(m,1H,CH),δ2.09(m,1H,CH),δ1.94(m,1H,CH),δ1.9-0.92(m,29H),δ0.91(s,3H,C
19H),δ0.64(s,3H,C
18H).
13C?NMR(400MHz,CDCl
3)δ:173.69，145.15，123.17，82.24，80.52，71.88，56.48，55.97，50.67，42.74，42.10，40.43，40.17，35.85，35.48，35.35，34.82，34.58，33.42，31.62，30.55，28.25，27.19，26.41，24.21，23.38，20.82，18.37，12.03
Embodiment 3
Click mark synthon 2-nitrine-1-[
18f] Radio-synthesis of fluoroethane, the putting productive rate of this step can reach more than 70%.
[example one] 20mCi
18f
-by quaternary ammonium type anion post QMA(U.S. Waters company product,
18f
-by Shanghai, Ke Xing pharmaceutcal corporation, Ltd provides) catch after, getting 1.0mL K222(is Kryptofix222) solution (14.4mg K222,3.0mg K
2cO
3, 960 μ L acetonitriles, the solution that 40 μ L water are made into) and will
18f
-be flushed in reaction flask, reaction flask immerses the oil bath of 95 ℃, and nitrogen dries up, and then adds 500 μ L anhydrous acetonitriles to dry up, and repeats twice of aforesaid operations; By the anhydrous acetonitrile of 400 μ L2-azidoethyl p-toluenesulfonic esters (5mg), under nitrogen protection, add rapidly reaction flask, confined reaction 5min at 95 ℃, stopped reaction, ice-water bath is cooling.Mark rate can reach more than 95%.
95 ℃ of auxiliary distillations of nitrogen, condensed fluid collection, in the receiving flask that contains 100 μ L acetonitriles, is added acetonitrile 100 μ L in reaction solution, continues distillation.Total distillation time 9-20min, distillation efficiency can reach 79%.
Containing 2-nitrine-1-[
18f] the acetonitrile condensation of fluoroethane collects liquid (approximately 600 μ L) and can be used in lower step mark reaction.
[example two], 30mCi
18f
-by quaternary ammonium type anion post QMA(U.S. Waters company product,
18f
-you Kexing pharmaceutcal corporation, Ltd provides) catch after, get 1.2mL K
222(being Kryptofix222) solution (17.5mg K
222, 3.5mg K
2cO
3, 1152 μ L acetonitriles, the solution that 48 μ L water are made into) and will
18f is flushed in reaction flask, and reaction flask immerses the oil bath of 95 ℃, and nitrogen dries up, and then adds 500 μ L anhydrous acetonitriles to dry up, and repeats anhydrous acetonitrile and dries up twice; Then 5mg2-azidoethyl p-toluenesulfonic esters is dissolved in to 500 μ L anhydrous acetonitriles, under nitrogen protection, adds rapidly reaction flask, confined reaction 10min at 95 ℃, stopped reaction, ice-water bath is cooling.Mark rate can reach more than 97%.
In reaction solution, add acetonitrile 200 μ L, nitrogen is assisted current-carrying, and distill, and collect phlegma, distillation 10-20min, distillation efficiency can reach 75%.
Containing 2-nitrine-1-[
18f] the acetonitrile condensation of fluoroethane collects liquid (approximately 700 μ L) and can be used in lower step mark reaction.
The nuclear magnetic data of 2-nitrine-1-fluoroethane is as follows:
1H?NMR(300MHz,CDCl
3)δ:4.685～4.498(dt,2H,CH
2-F,
2J
FH=47Hz,?
2J
HH=4.5Hz);3.579～3.456(dt,2H,CH
2-CH
2-F,
3J
FH=27Hz,
2J
HH=4.5Hz).
The radio-labeling of bile acids compound
[example one], the radio-labeling to stone courage acyl propargylamine (B6)
In 200 μ L pH=6.0 phosphate buffered saline buffers, successively add 60 μ L copper-baths (0.45mol/L) and 60 μ L sodium ascorbate solutions (1.5mol/L), the DMF solution (400 μ L) that adds stone courage acyl propargylamine (B6) 4.0mg after mixing, after vortex mixed 1min, the 2-nitrine-1-[that adds distillation condensation to obtain
18f] acetonitrile solution (V=300 μ L, 1mCi) of fluoroethane, 50 ℃ of reaction 15min of vibrator.
Marked product detects also separation with HPLC, and (U.S. Agilent1100HPLC system, semipreparative column is Agilent C18column (9.3mm * 250mm).Moving phase is to have added water (X) and the acetonitrile (Y) of 0.1% trifluoroacetic acid, and gradient separations condition is: 0-25min, 30%Y → 70%Y.Flow velocity is 4.0mL/min.Through UV(263nm) detect and radioassay.) collection product peak (t
r=23.8min), concentrating under reduced pressure is removed solvent, with dissolving containing the PBS (pH=7.4) of 8% ethanol or containing 0.9% medical saline of 8% ethanol, obtains the preparation of A6-second after aseptic membrane filtration.
Same A ' the 6-of the appraising datum second of A6-second.
[example two], the radio-labeling to CDCA acyl propargylamine (B3)
In 200 μ L pH=6.0 phosphate buffered saline buffers, successively add 60 μ L copper-baths (0.45mol/L) and 60 μ L sodium ascorbate solutions (1.5mol/L), the DMF solution (400 μ L) that adds CDCA acyl propargylamine (B3) 4.0mg after mixing, after vortex mixed 1min, the 2-nitrine-1-[that adds distillation condensation to obtain
18f] acetonitrile solution (V=300 μ L, 1.5mCi) of fluoroethane, 50 ℃ of reaction 15min of vibrator.
Marked product detects also separation with HPLC, and (U.S. Agilent1100HPLC system, semipreparative column is Agilent C18column (9.3mm * 250mm).Moving phase is to have added water (X) and the acetonitrile (Y) of 0.1% trifluoroacetic acid, and gradient separations condition is: 0-10-20-25min, 40%Y → 65%Y → 55%Y → 40%Y.Flow velocity is 2.0mL/min.Through UV(205nm) detect and radioassay.) collection product peak (t
r=14.3min), concentrating under reduced pressure is removed solvent, with dissolving containing the PBS (pH=7.4) of 8% ethanol or containing 0.9% medical saline of 8% ethanol, obtains the preparation of A3-second after aseptic membrane filtration.
Same A ' the 3-of the appraising datum second of A3-second.
The vitro stability of tagged compound is measured
The vitro stability of the marked product A6-second of the example one of [example one], embodiment 4 is measured
1. the stability in PBS: get
18 f mark preparation 50 μ L(approximately 60 μ Ci), be placed in 1mL PBS, be placed at 37 ℃, after oscillation incubation 30,60,120,240,360min, with HPLC, measure its radiochemical purity, to observe its vitro stability.HPLC analytical results shows that the tagged compound of this embodiment produces there are no radioimpurity radioactive impurity, shows that it has satisfactory stability in PBS, as shown in Figure 1.
2. the stability in serum: get
18 f mark preparation 50 μ L(approximately 80 μ Ci), be placed in 1000uL calf serum, be placed at 37 ℃, after oscillation incubation 30,60,120,240,360min, sample 100 μ L and be placed in the dactylethrae that contains 200 μ L methyl alcohol, mix latter centrifugal two minutes, get supernatant liquor and measure its radiochemical purity with HPLC, to observe its stability in serum.HPLC analytical results shows that the tagged compound of this embodiment produces there are no radioimpurity radioactive impurity, shows that it has satisfactory stability in calf serum, as shown in Figure 2.
The vitro stability of the marked product A3-second of the example two of [example two], embodiment 4 is measured
1. the stability in PBS: get
18 f mark preparation 50 μ L(approximately 80 μ Ci), be placed in 1mL PBS, be placed at 37 ℃, after oscillation incubation 30,60,120,240,360min, with HPLC, measure its radiochemical purity, to observe its vitro stability.HPLC analytical results shows that the tagged compound of this embodiment produces there are no radioimpurity radioactive impurity, shows that it has satisfactory stability in PBS, as shown in Figure 4.
2. the stability in serum: get
18 f mark preparation 50 μ L(approximately 80 μ Ci), be placed in 1000uL calf serum, be placed at 37 ℃, after oscillation incubation 30,60,120,240,360min, sample 100 μ L and be placed in the dactylethrae that contains 200 μ L methyl alcohol, mix latter centrifugal two minutes, get supernatant liquor and measure its radiochemical purity with HPLC, to observe its stability in serum.HPLC analytical results shows that the tagged compound of this embodiment produces there are no radioimpurity radioactive impurity, shows that it has satisfactory stability in calf serum, as shown in Figure 3.
Embodiment 6
The mensuration of the lipotropy of tagged compound (Log P)
The lipotropy of tagged compound is measured in the system of n-Octanol and water-soluble buffered soln (PBS) composition.This index is to weigh an important factor of medicine absorption, distribution, metabolism and elimination in vivo.
Log P is the lipid of compound, can reflect a lipophilic power of compound.It measures calculation formula:
Get 10 μ L
18f mark preparation (contains 600 μ L n-Octanols and 590 μ LPBS) in 2mL dactylethrae, good seal, and at room temperature, after abundant vortex 2min, high speed centrifugation 3min, to biphase equilibrium.With pipettor, from organic phase and water, respectively sample 100 μ L and be placed in two V counter tubes, with gamma counter, measure counting.Repeated sampling is measured three times.Parallelly carry out two batches of experiments, calculate
18the average Log P value of F mark preparation.
The mensuration of the lipotropy (Log P) of the tagged compound A6-second of the example one of [example one], embodiment 4
According to twice parallel laboratory test, three groups of panel datas that at every turn record, calculate Log P=1.78.Lipotropy data presentation, tagged compound has good lipotropy, is easy to enter liver, passes through liver metabolism.
The mensuration of the lipotropy (Log P) of the tagged compound A3-second of the example two of [example two], embodiment 4
According to twice parallel laboratory test, three groups of panel datas that at every turn record, calculate Log P=1.61.Lipotropy data presentation, tagged compound has good lipotropy, is easy to enter liver, passes through liver metabolism.
Embodiment 7
The application of the preparation of the tagged compound A3-second of the example two of embodiment 4 in mouse PET-CT video picture
After mouse anesthesia, carry out behind CT scan location,
18f mark preparation A3-second 100 μ L(approximately 50 μ Ci) by tail vein injection in Mice Body, Mobile state PET.In whole scanning process, guarantee the Oxygen Flow of 2L/min, be mixed with 1.5% isoflurane.Inveon Acquisition Workplace (IAW) controls whole scanning process.Utilize OSEM3D (Three-Dimensional Ordered Subsets Expectation Maximum) algorithm to rebuild image.
Dynamically PET-CT video picture shows: tracer agent (
18f mark preparation) A3-second by tail vein injection to after in Mice Body, liver organization is picked-up tracer agent rapidly, during 1min, a large amount of tracer agent (shown in Fig. 5) has concentrated at liver position, tracer agent is secreted into stones in intrahepatic bile duct and total hepatic duct subsequently, by common bile duct, be secreted into enteron aisle, remaining tracer agent is in gall-bladder accumulation (shown in Fig. 6).Finally, tracer agent, substantially from hepatic clearance, is mainly distributed to enteron aisle or concentrates at gall-bladder (shown in Fig. 7), and tracer agent is 1h in mouse body, still finds no defluorinate video picture, has good stability.
Fig. 8 is tracer agent A3-second at the SUV-t of the liver of normal mice curve (SUV refer to radioactive activity and the whole body of the tracer agent of local organization picked-up on average inject the ratio of activity), this curve shows the metabolic process removed from concentrating at liver of tracer agent, as can be seen from the figure, half clean-up time of tracer agent in Mice Body is 25min, and this shows that tracer agent has the interior removing speed of body faster.
Claims (15)
1. suc as formula the bile acids compound shown in A,
Wherein, R
1, R
2and R
3be H or OH independently of one another; R
4for methylene radical,
or
described
in the carbon that is connected with carbonyl with in described A-NH is connected, described
in the ethylidene that is connected with sulfuryl with in described A-NH is connected; R
5for C
1～C
10straight-chain alkyl-sub-or C
1～C
10branched alkylidene, C
5～C
7cycloalkylidene, poly-sub-ethylene glycol, C
6～C
10arylidene, C
6～C
10arylidene-C
1～C
4alkylidene group or C
1～C
6alkylidene group-C
6～C
10arylidene-C
1～C
4alkylidene group, described C
6～C
10arylidene-C
1～C
4in alkylidene group-C
1～C
4alkylidene group is connected with the fluorine in described A;
Described poly-sub-ethylene glycol is following group:
N2=2～7 wherein.
2. bile acids compound as claimed in claim 1, is characterized in that, described F is
18f.
3. bile acids compound as claimed in claim 1, is characterized in that, described C
1～C
10straight-chain alkyl-sub-be following group:
Described C
1～C
10branched alkylidene be following group:
N1=0～8 wherein.
4. bile acids compound as claimed in claim 3, is characterized in that, n1 is 1,2 or 3.
5. bile acids compound as claimed in claim 1, is characterized in that, n2 is 2 or 3.
6. bile acids compound as claimed in claim 1, is characterized in that, described C
6～C
10arylidene is
7. bile acids compound as claimed in claim 1, is characterized in that, described C
6～C
10arylidene-C
1～C
4alkylidene group is following group:
8. bile acids compound as claimed in claim 1, is characterized in that, described C
1～C
6alkylidene group-C
6～C
10arylidene-C
1～C
4alkylidene group is following arbitrary group:
n3=1～5，n4=0～4。
9. bile acids compound as claimed in claim 1, is characterized in that, described C
5～C
7cycloalkylidene is following group:
N5=1～3 wherein.
10. bile acids compound as claimed in claim 1, is characterized in that, described A is,
A1: work as R
1=-H, R
2=-α OH, R
3=-α OH, R
4=
time compd A;
Or, A2: work as R
1=-H, R
2=-H, R
3=-α OH,
time compd A;
Or, A3: work as R
1=-H, R
2=-α OH, R
3=-H,
time compd A;
Or, A4: work as R
1=-H, R
2=-β OH, R
3=-H,
time compd A;
Or, A5: work as R
1=-α OH, R
2=-H, R
3=-H,
time compd A;
Or, A6: work as R
1=-H, R
2=-H, R
3=-H,
time compd A;
Or, A7: work as R
1=-H, R
2=-α OH, R
3=-α OH,
time compd A;
Or, A8: work as R
1=-H, R
2=-H, R
3=-α OH,
time compd A;
Or, A9: work as R
1=-H, R
2=-α OH, R
3=-α OH,
time compd A;
Or, A10: work as R
1=-H, R
2=-α OH, R
3=-α OH,
time compd A;
Or, A11: work as R
1=-H, R
2=-H, R
3=-α OH,
time compd A;
Or, A12: work as R
1=-H, R
2=-α OH, R
3=-H,
time compd A.
The preparation method of 11. bile acids compounds as described in any one in claim 1～10, it is characterized in that, it comprises the steps: in solvent, under the catalysis of Cu (I), compd B and Compound D are carried out to nitrine and the Huisgen1 that holds position alkynyl, 3-Dipolar Cycloaddition;
Wherein, R
1, R
2, R
3, R
4and R
5definition all with described in any one in claim 1～10.
12. 1 kinds of compd Bs,
Wherein, R
1, R
2and R
3be H or OH independently of one another, R
4for methylene radical,
or
13. compd Bs as claimed in claim 12, is characterized in that, compd B is,
B1: work as R
1=-H, R
2=-α OH, R
3=-α OH,
time compd B;
Or, B2: work as R
1=-H, R
2=-H, R
3=-α OH,
time compd B;
Or, B3: work as R
1=-H, R
2=-α OH, R
3=-H,
time compd B;
Or, B4: work as R
1=-H, R
2=-β OH, R
3=-H,
time compd B;
Or, B5: work as R
1=-α OH, R
2=-H, R
3=-H,
time compd B;
Or, B6: work as R
1=-H, R
2=-H, R
3=-H,
time compd B;
Or, B7: work as R
1=-H, R
2=-α OH, R
3=-α OH,
time compd B;
Or, B8: work as R
1=-H, R
2=-H, R
3=-α OH,
time compd B;
Or, B9: work as R
1=-H, R
2=-α OH, R
3=-α OH,
time compd B;
Or, B10: work as R
1=-H, R
2=-α OH, R
3=-α OH,
time compd B;
Or, B11: work as R
1=-H, R
2=-H, R
3=-α OH,
time compd B;
Or, B12: work as R
1=-H, R
2=-α OH, R
3=-H,
time compd B.
14. 1 kinds of arbitrary Compound C 10-1, C10-2, C10-3, C11-1, C11-2, C11-3, C12-1, C12-2 or C12-3 as follows,
15. 1 kinds of bile acids compounds as described in any one in claim 1～10 are as the application of positron emission tomography agent, and wherein the F in compd A is
18f.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310365746.8A CN103570793B (en) | 2013-08-20 | 2013-08-20 | Bile acids compound, intermediate and its preparation method and application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310365746.8A CN103570793B (en) | 2013-08-20 | 2013-08-20 | Bile acids compound, intermediate and its preparation method and application |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103570793A true CN103570793A (en) | 2014-02-12 |
| CN103570793B CN103570793B (en) | 2015-08-12 |
Family
ID=50043569
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310365746.8A Active CN103570793B (en) | 2013-08-20 | 2013-08-20 | Bile acids compound, intermediate and its preparation method and application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103570793B (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101891791A (en) * | 2009-05-22 | 2010-11-24 | 中国科学院上海应用物理研究所 | Derivate for labeling bile acid and reference compound, preparation method and application thereof |
| CN102351941A (en) * | 2011-06-08 | 2012-02-15 | 中国科学院上海应用物理研究所 | Method for labeling functional molecules with <18>F |
-
2013
- 2013-08-20 CN CN201310365746.8A patent/CN103570793B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101891791A (en) * | 2009-05-22 | 2010-11-24 | 中国科学院上海应用物理研究所 | Derivate for labeling bile acid and reference compound, preparation method and application thereof |
| CN102351941A (en) * | 2011-06-08 | 2012-02-15 | 中国科学院上海应用物理研究所 | Method for labeling functional molecules with <18>F |
Non-Patent Citations (2)
| Title |
|---|
| NAMDEV S. VATMURGE ET AL.: "Synthesis and antimicrobial activity of β-lactam–bile acid conjugates linked via triazole", 《BIOORGANIC & MEDICINAL CHEMISTRY》 * |
| NAMDEV S. VATMURGE ET AL.: "Synthesis and biological evaluation of bile acid dimers linked with 1,2,3-triazole and bis-b-lactam", 《ORGANIC & BIOMOLECULAR CHEMISTRY》 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103570793B (en) | 2015-08-12 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106967152B (en) | A kind of compound and the preparation method and application thereof of Value linear label | |
| Fujinaga et al. | Synthesis and evaluation of 6-[1-(2-[18F] fluoro-3-pyridyl)-5-methyl-1H-1, 2, 3-triazol-4-yl] quinoline for positron emission tomography imaging of the metabotropic glutamate receptor type 1 in brain | |
| Liu et al. | A new 18 F-heteroaryltrifluoroborate radio-prosthetic with greatly enhanced stability that is labelled by 18 F–19 F-isotope exchange in good yield at high specific activity | |
| CN111138504B (en) | A kind of99mTc-CNPEDG complex and preparation method and application thereof | |
| CN112175025B (en) | Glucose derivative containing benzene ring and application thereof | |
| CN115010629B (en) | Prostate specific membrane antigen inhibitor, nuclide marker, preparation method and application | |
| Xu et al. | 18F–labeled estradiol derivative for targeting estrogen receptor-expressing breast cancer | |
| CN115974962A (en) | FAP (FAP-associated protein) targeted probe as well as preparation method and application thereof | |
| Shi et al. | [68Ga] Ga-HBED-CC-DiAsp: A new renal function imaging agent | |
| CN101768208B (en) | Novel 18F-labelled polypeptide, preparation method and application thereof in tumor imaging | |
| CN101723850B (en) | Novel 18F labeled aromatic amino acids, preparation method and application thereof in tumor imaging | |
| CN117209476A (en) | The method comprises the following steps of 99m Tc-labeled radioactive probe for targeting fibroblast activation protein and preparation method and application thereof | |
| CN113583066B (en) | Mannose derivative and application thereof | |
| CN103570793B (en) | Bile acids compound, intermediate and its preparation method and application | |
| Jain et al. | A systematic comparative evaluation of 68 Ga-labeled RGD peptides conjugated with different chelators | |
| Kong et al. | Development of Tyrosine‐Based Radiotracer 99mTc‐N4‐Tyrosine for Breast Cancer Imaging | |
| JP6037330B2 (en) | 11C-labeled thiamine and derivatives thereof, 11C-labeled fursultiamine, thiamine precursor, and probe for PET and imaging method using them | |
| Yang et al. | Synthesis and biological evaluation of 99mTc-DMP-NGA as a novel hepatic asialoglycoprotein receptor imaging agent | |
| Yang et al. | Synthesis and evaluation of 111 In-labeled d-glucose as a potential SPECT imaging agent | |
| Liu et al. | Synthesis and biological evaluation of novel technetium-99m-labeled HYNIC-D-glucose as a potential tumor imaging agent | |
| CN105985406B (en) | A kind of18F-labeled polypeptide compound and preparation method and application thereof | |
| Farn et al. | Synthesis, radiolabeling, and preliminary in vivo evaluation of [68ga] ipcat-nota as an imaging agent for dopamine transporter | |
| CN110577478A (en) | Positron probe and preparation method and application thereof | |
| CN115368342B (en) | Fibroblast active protein inhibitor, radionuclide marker, preparation method and application thereof | |
| Brown et al. | Radiosynthesis and analysis of (S)-4-(3-[18f] Fluoropropyl)-L-Glutamic acid |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20200512 Address after: 201815 Four and Six Frost Bamboo Highway 4288, Jiading District, Shanghai Patentee after: Shanghai Shenjing Pharmaceutical Technology Co.,Ltd. Address before: 201800 Shanghai city Jiading District Baojia Highway No. 2019 Patentee before: SHANGHAI INSTITUTE OF APPLIED PHYSICS, CHINESE ACADEMY OF SCIENCES |
|
| TR01 | Transfer of patent right |