CN110698515B

CN110698515B - Marked fatty acid derivative containing isocyano group and application thereof

Info

Publication number: CN110698515B
Application number: CN201811364458.XA
Authority: CN
Inventors: 张华北; 强兵朝
Original assignee: Beijing Normal University
Current assignee: Beijing Normal University
Priority date: 2018-07-31
Filing date: 2018-11-16
Publication date: 2021-03-26
Anticipated expiration: 2038-11-16
Also published as: CN110698515A

Abstract

The invention relates to a marked fatty acid derivative containing isocyano and application thereof, wherein the general formula is shown as formula I, wherein M is marked technetium^99mTc or rhenium Re; r, R1, R2, R3, R4 and R5 are all H, aliphatic chains or alicyclic chains, and the five groups are completely identical, completely different or partially identical; z is an integer from 1 to 6, p is 0 or 1, n is a positive integer from 1 to 28; t is an integer of 0 to 5, and a is an integer of 0 to 5. The invention provides a completely novel structure type, which has lower liver background, lung background and kidney background, but has long detention time and high detention rate in cardiac muscle, can be used as a novel cardiac muscle metabolism developer, and has important clinical application prospect.

Description

Marked fatty acid derivative containing isocyano group and application thereof

Technical Field

The invention relates to the technical field of medicine and chemistry, in particular to a marked fatty acid derivative containing isocyano and application thereof.

Background

Fatty acids are the simplest of the lipids, and are a constituent of many more complex lipids. Fatty acids can be oxidatively decomposed to CO with sufficient oxygen supply₂And H₂O, releases a large amount of energy, and thus fatty acids are one of the main energy sources of the body. Especially the main energy source for myocardial contraction.

Action of fatty acids in the cardiac muscle the process of muscle contraction requires higher energy and therefore there must be continuous, efficient ATP to maintain contractile function, basal metabolism and ion exchange balance. In normal human myocardium, fatty acid oxidation provides 60% -80% of the energy required by myocardium, and in addition, myocardium can also obtain the energy required for physiological activities by oxidizing energy substances such as glucose, pyruvic acid, amino acids, lactic acid, and the like. When the organism is at a low blood glucose level or is fasting, the heart has a high uptake of fatty acids, and the oxygen consumption of the myocardium is almost always available for the oxidation of fatty acids.

When the oxygen supply to the cardiac muscle is sufficient, the fatty acid can efficiently provide energy to the cardiac muscle through beta-oxidation; when myocardial ischemia or oxygen supply is low, beta-oxidation of fatty acid is inhibited, glucose metabolism is changed into energy for myocardial metabolism, and utilization rate of fatty acid by myocardium is reduced.

The metabolic pathway of myocardial fatty acid imaging agents is substantially similar to free fatty acids. The oxidation of fatty acids is divided into 3 steps, the first step is the activation of fatty acids in the cell fluid, the fatty acids are catalyzed by fatty acyl-CoA synthetase to produce fatty acyl-CoA, the second step is the transport of fatty acyl-CoA into mitochondria, and the third step is the β -oxidation of fatty acyl-CoA. Long chain fatty acids undergo a single beta-oxidation to remove 2 carbons of acetyl CoA from the beta carbon to form two fewer carbons of acyl CoA, which releases a large amount of energy.

The beta-oxidation of fatty acyl CoA in the myocardium is divided into 4 steps, which are respectively removing one H from alpha and beta positions, adding water, dehydrogenating to generate beta-fatty acyl CoA and thiolating, wherein each step corresponds to different enzymes: acyl-CoA dehydrogenase, enoyl-CoA hydratase, 3-OH acyl-CoA dehydrogenase, beta-ketoacyl-CoA thiolase.

At present, clinically used myocardial metabolism imaging agents mainly include: [11C ] -palmitate and [18F ] -FDG for PET imaging, and [123I ] -IPPA and [123I ] BMIPP for SPECT imaging. The half-life of 11C is only 110min, the requirements on labeling and imaging speed are high, the price of 18F-FDG is high, radioiodinated drugs are easy to deiodinate in vivo, radioiodinated drugs need to be purchased from special companies, and the defects which cannot be overcome by the radioiodinated drugs are overcome by the different drugs. The 99mTc has excellent nuclide property, has a half-life of 6h, is convenient for labeling, transporting and using drugs, is low in price and convenient to obtain, and has the other important advantage that the 99mTc has different valences (+1 to +7) and can form various different coordination structures, and the coordination number is 5, 6 and 7. Among them, there is a labeling method which has realized kit formation, and is convenient for clinical application. However, no 99mTc labelled imaging agent of myocardial metabolism has been successfully used in the clinic.

In 2008, Byung Chul Lee and the like introduce carbonyl on the basis of the structure of a compound [99mTc ] CpTT-PA, and the compound 99mTc-CpTT-16-oxo-HAD is designed and synthesized, so that the hydrophilicity is increased, and the liver uptake is reduced. The compound is metabolized in a beta-oxidation mode in the myocardium, and finally the compound 99mTc-CpTT-4-oxo-butyric acid is generated through metabolism, the myocardial uptake value is higher than that of the compound [99mTc ] CpTT-PA, but the liver background, the lung background and the kidney background are higher, and an ideal target-to-non-target ratio is not obtained.

In 2012, the great huahui and the like introduce amide bonds on the basis of the long chain structure of the compound 99mTc-CpTT-16-oxo-HAD to synthesize the compound 99mTc-CpTT-6-oxo-HAUA, the compound is reduced in liver, but the myocardial uptake value is low, and the myocardial uptake is only 4.37% ID/g in 1 min.

Disclosure of Invention

In view of the above, the present invention aims to provide a labeled isocyano group-containing fatty acid derivative and an application thereof, which solve the disadvantages in the prior art.

The purpose of the invention is realized by the following technical scheme:

a labeled isocyano group-containing fatty acid derivative has a general formula shown in formula I:

wherein M is labelled technetium^99mTc or labeled rhenium Re;

r, R1, R2, R3, R4 and R5 are all H, aliphatic chains or alicyclic chains, and the five groups are completely identical, completely different or partially identical;

z is an integer of 1-6, 6-z is 6 minus z, p is 0 or 1, n is a positive integer of 1-28; t is an integer of 0 to 5, and a is an integer of 0 to 5.

Further, when p is 1 and a and t are both 1, formula I is represented by the following general formula II:

when p is 0, a and t are both 1, formula I is of the following formula III:

the application of labeled fatty acid derivative containing isocyano group in myocardial imaging agent.

The invention provides a marked fatty acid derivative containing isocyano and application thereof, which mainly have the following effects: the invention provides a completely novel structure type, namely when at least one ligand end in the novel structure is carboxylic acid or ester group, namely z is not equal to 0, the compound has high uptake and long retention time in cardiac muscle, can be used as a cardiac muscle fatty acid metabolism imaging agent, and has important clinical application prospect.

Drawings

The spectrum referred to in the present invention will be briefly described as follows.

FIG. 1 is a mass spectrum of ligand 1 according to example 4 of the present invention;

FIG. 2 is a mass spectrum of the product I-1-Re of example 3 of the present invention;

FIG. 3 is an HPLC plot of the product I-1-Re of example 3 of the present invention;

FIG. 4 is a mass spectrum of I-2-Re of example 3 of the present invention;

FIG. 5 is an HPLC plot of I-2-Re of example 3 of the present invention;

FIG. 6 shows I-1-^99mHPLC plot of Tc;

FIG. 7 shows I-2-^99mHPLC plot of Tc;

FIG. 8 is a mass spectrum of I-3-Re of example 5 of the present invention;

FIG. 9 is a mass spectrum of I-4-Re of example 5 of the present invention;

FIG. 10 is an HPLC plot of I-4-Re of example 5 of the present invention;

FIG. 11 shows I-4-^99mHPLC plot of Tc;

FIG. 12 is a mass spectrum of I-5-Re of example 6 of the present invention;

FIG. 13 is an HPLC chromatogram of I-5-Re of example 6 of the present invention;

FIG. 14 is an HPLC chromatogram of I-6-Re of example 6 of the present invention;

FIG. 15 is a mass spectrum of I-6-Re of example 6 of the present invention;

FIG. 16 shows I-5-^99mHPLC profile of Tc;

FIG. 17 shows I-6-^99mHPLC profile of Tc;

FIG. 18 is an HPLC chromatogram of I-3-Re of example 5 of the present invention;

FIG. 19 shows I-3-^99mLiquid phase diagram of Tc.

Detailed Description

The detailed description of the embodiments of the present invention provided below is not intended to limit the scope of the invention as claimed, but is merely representative of selected embodiments of the invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

Example 1

wherein M is labelled technetium^99mTc or labeled rhenium Re; r, R1, R2, R3, R4 and R5 are all H, aliphatic chains or alicyclic chains, and these five groups are completely the same, completely different or partially the same groups.

z is an integer from 1 to 6, 6-z is 6 minus z, for example: if z is 2, 6-z is 4; p is 0 or 1, n is a positive integer from 1 to 28; t is an integer of 0 to 5, and a is an integer of 0 to 5.

Preferably, R, R1, R2, R3, R4 and R5 are all H or aliphatic chains, preferably aliphatic hydrocarbons, and more preferably aliphatic hydrocarbons of 1-28C.

Preferably, a and t are integers from 1 to 3, more preferably: a and t are both 1.

Preferably, n is a positive integer from 1 to 15, such as 1, 3, 5, 7, 9, 11, 13, 15, and the like.

Preferably, if p is 1 and a and t are both 1, formula I is of the following general formula II:

when p is 0, a and t are both 1, formula I is of the following formula III:

more preferably, if p is 1 and a and t are both 1, formula I includes, but is not limited to, the following compounds:

if p is 0 and a and t are both 1, formula I includes, but is not limited to, the following compounds:

wherein M is labelled technetium^99mTc, R, R1, R2, R3, R4 and R5 are all H or aliphatic hydrocarbons of 1-4C, and n is a positive integer of 1-28.

In the present embodiment, R is more preferably H, CH₃Or CH₂CH₃R1, R2, R3, R4 and R5 are all more preferably methyl CH₃

Example 2

The synthetic route and method for selecting the compound of example 1 is as follows:

1. synthesis of ligand 1

The synthesis route is as follows:

where R6 is the same as R in example 1, and the parameters of n and a are the same as in example, and the description thereof will not be repeated.

The synthesis process is as follows:

synthesis of Compound 1: adding 30mL (0.37mol) of ethyl formate into a round-bottom flask, slowly dropwise adding 0.07mol of the raw material 1 by using a constant-pressure dropping funnel, heating to 60 ℃ for 36h, and then removing the solvent by spinning off to obtain 6.78g of a product with the yield of 97.3%.

Synthesis of Compound 3: a round-bottomed flask was charged with 7.96mmol of Compound 2, 16mL of ethanol, and 3.79g (31.84mmol) of SOCl was added dropwise from a constant pressure dropping funnel at room temperature₂Then the temperature is increased and the reflux is carried out for 8 hours. Removing solvent, adding small amount of water, extracting with dichloromethane twice, mixing organic phases, and adding anhydrous NaSO₄Drying, filtering and removing the solvent by spinning to obtain the product.

Synthesis of Compound 4: 7.96mmol of compound 3 are placed in a round-bottom flask and placed in an ice bath, 31.84mmol of formic acid are added and triethylamine is slowly added dropwise. After the addition, the mixture was heated to 50 ℃ to react overnight. Adding brine, extracting with ethyl acetate, washing the organic phase with water, washing with 1mol/L hydrochloric acid, washing with saturated sodium carbonate, and washing with brine. After drying the sodium sulfate, the solvent is removed by rotation to obtain a crude product which is directly subjected to the next reaction. Adding methanol and 1 drop of concentrated hydrochloric acid, reacting at room temperature for 2h, removing most of methanol by spinning, adding a small amount of water and ethyl acetate for extraction, washing with saturated sodium carbonate, washing with water and brine to obtain an organic phase, drying, spinning off the solvent, and purifying by TLC to obtain the product.

Synthesis of Compound 5: a round-bottomed flask was charged with 1.02g (10.33mmol) of Compound 1, 10mL of anhydrous dichloromethane, 10.33mmol of Compound 4, 12.8mol of Mercury acetate were added, the mixture was stirred at room temperature for 14 hours, filtered, the solvent was removed by spinning, the mixture was dissolved in dichloromethane, and 10.84mmol of NaBH was added slowly in portions₄Stirring was continued for 2h and then filtered. And (5) removing the solvent from the filtrate by spinning to obtain the product.

Synthesis of ligand 1: adding 10.33mmol of compound 5 into a round-bottom flask, adding 10mL of anhydrous dichloromethane and 10.33mmol of triethylamine, slowly dropwise adding 4.08g (12.8mol) of phosphorus oxychloride, stirring at normal temperature for 6h to stop the reaction after the dropwise adding is finished, adding 80mL of 20% sodium carbonate aqueous solution, stirring for 45min, extracting with dichloromethane twice, combining organic phases, washing with brine and anhydrous MgSO₄And (5) drying. Filtering, selecting and adjusting the solvent to obtain a crude product, and purifying the crude product by a liquid phase.

2. Synthesis of ligand 2

The synthesis route is as follows:

wherein R5 is the same as R5 in example 1, and the t parameter is the same as in example, and the description thereof is not repeated.

The synthesis process comprises the following steps:

Synthesis of ligand 2: adding 10.33mmol of compound 6 into a round-bottom flask, adding 10mL of anhydrous dichloromethane and 10.33mmol of triethylamine, slowly dropwise adding 4.08g (12.8mol) of phosphorus oxychloride, stirring at normal temperature for 6h to stop the reaction after the dropwise adding is finished, adding 80mL of 20% sodium carbonate aqueous solution, stirring for 45min, extracting with dichloromethane twice, combining organic phases, washing with brine and anhydrous MgSO₄And (5) drying. Filtering, selecting and adjusting the solvent to obtain a crude product, and purifying the crude product by a liquid phase.

3. Synthesis of ligand 3

The synthesis route is as follows:

the synthesis process comprises the following steps:

20ml of methanol was added to a round-bottom flask, 5mmol of Compound 1 was dissolved in methanol, then 20ml of ethyl formate and 750. mu.l of triethylamine were added, refluxed for 36 hours, the solvent was removed by spinning, the residue was dissolved in dichloromethane, washed three times with water, and the oil phase was spin-dried to give colorless oil 2.

Compound 2(3.0mol) was dissolved in 15ml of methylene chloride, and 15.0mol of triethylamine was added. 3.3mmol of POCl was added dropwise at 0 deg.C₃At 30min, room temperature for 3h and then saturated NaHCO₃Quenching withMgSO water₄Drying, filtering, concentrating, and purifying with flash silica gel column (mobile phase is dichloromethane) to obtain compound 3.

4. Synthesis of the general formula II

If M is^99mWhen Tc, the synthesis method is as follows: ligand 1 and ligand 2 were dosed in a molar ratio [ i.e., ligand 1: ligand 2 ═ m: (6-m)]A total of about 1-5mg, was dissolved in 700 μ L of physiological saline in a 10ml vial, and then 200 μ L of CB (citrate buffer, 0.1m, pH 6.6) and 20 μ L of sncl2h2O solution (1 mg of sncl2h2O/1 ml) were added to the vial. After addition of 100. mu.l of a 99 MTcCO 4-containing brine (370-3700Mbq), the solution was heated at 100 ℃ for 30 minutes. Extracting with dichloromethane, purging with nitrogen to remove dichloromethane, dissolving in 200 μ L methanol, treating with 0.3N NaOH 50 μ L, reacting at 70C for 10min, cooling, acidifying with 0.1N HCl 150 μ L, extracting with dichloromethane, and purifying with HPLC (water: methanol: 8:2) to obtain the final product^99mTc-labelled formula II.

If M is rhenium, the synthesis method is as follows: two ligands, NH₄ReO₄And anhydrous stannous chloride in water, adjusting the pH to 6 with gentisic acid, boiling for 30 minutes, extracting with dichloromethane, and removing dichloromethane to obtain the general formula ii, and obtaining the ester, i.e. R is aliphatic.

The resulting ester was then dissolved in methanol and reacted with 0.3N NaOH at 70 ℃ for 10min, cooled and acidified to pH 2 with 0.1N HCl, extracted with dichloromethane and purified by HPLC (water: methanol 8:2) to give formula ii and the acid, i.e. R ═ H.

5. Synthesis of formula III

Ligand 2 and ligand 3 were dosed in molar ratio [ i.e., ligand 2: ligand 3 ═ (6-m): m ], the specific procedure is identical to that of the synthesis of formula II and is not repeatedly defined herein.

Example 3

With M ═ Re and^99mtc, m-1, n-5 were used as examples to synthesize the following compounds I-1-Re (ester), I-2-Re (acid), I-1-^99mTc (ester) and I-2-^99mTc (acid).

Synthetic route to ligand 1:

synthesis of compound 1: adding 30mL (0.37mol) of ethyl formate into a round-bottom flask, slowly dropwise adding 5g (0.07mol) of 2-methylacrylamine (raw material 1) by using a constant-pressure dropping funnel, heating to 60 ℃, heating for 36h, and then removing the solvent to obtain 6.78g of a product with the yield of 97.3%.

Synthesis of Compound 3: a round-bottomed flask was charged with 1.56g (7.96mmol) of 6-bromohexanoic acid (i.e., Compound 2) and 16mL of ethanol, and 3.79g (31.84mmol) of SOCl was added dropwise from an isopiestic dropping funnel at room temperature₂Then the temperature is increased and the reflux is carried out for 8 hours. Removing solvent, adding small amount of water, extracting with dichloromethane twice, mixing organic phases, and adding anhydrous NaSO₄Drying, filtering and removing the solvent by spinning to obtain 1.59g of the product, namely the compound 3, with the yield of 89.2%.

Synthesis of Compound 4: a round-bottomed flask was charged with 1.56g (7.96mmol) of ethyl 6-bromohexanoate (Compound 3), placed in an ice bath, and after addition of 3.79g (31.84mmol) of formic acid triethylamine was slowly added dropwise. After the addition, the mixture was heated to 50 ℃ to react overnight. Adding brine, extracting with ethyl acetate, washing the organic phase with water, washing with 1mol/L hydrochloric acid, washing with saturated sodium carbonate, and washing with brine. Drying the sodium sulfate, and then removing the solvent by spinning to obtain a crude product which is directly subjected to the next reaction; directly adding methanol and 1 drop of concentrated hydrochloric acid, reacting at room temperature for 2h, removing most of methanol by spinning, adding a small amount of water and ethyl acetate for extraction, washing with saturated sodium carbonate, washing with water and brine to obtain an organic phase, drying, spinning off a solvent, and purifying by TLC to obtain a product, namely the compound 4 with a yield of 82.2%.

Synthesis of Compound 5: a round-bottomed flask was charged with 1.02g (10.33mmol) of Compound 1, 10mL of anhydrous dichloromethane and 1.80g (10)33mmol) 6-hydroxyacetic acid ethyl ester (Compound 4), 4.08g (12.8mol) mercuric acetate, stirring at room temperature for 14h, filtering, removing the solvent by spinning, dissolving in dichloromethane, slowly adding 0.41g (10.84mmol) NaBH in portions₄Stirring was continued for 2h and then filtered. The solvent is removed from the filtrate by spinning to obtain the product, namely the compound 5 with the yield of 70 percent.

Synthesis of ligand 1: adding 1.02g (10.33mmol) of compound 5 into a round-bottom flask, adding 10mL of anhydrous dichloromethane and 1.80g (10.33mmol) of triethylamine, slowly dropwise adding 4.08g (12.8mol) of phosphorus oxychloride, stirring at normal temperature for 6h after the dropwise adding is finished, stopping the reaction, adding 80mL of 20% sodium carbonate aqueous solution, stirring for 45min, extracting with dichloromethane twice, combining organic phases, washing with brine, and washing with anhydrous MgSO₄And (5) drying. Filtering, selecting and adjusting a solvent to obtain a crude product, and purifying a liquid phase to obtain a product, namely the ligand 1, wherein the yield is 75%.

Ligand 2

The synthesis of (2): the synthesis method is the same as example 2 and is not described herein.

Synthesizing a target product I-1-Re: then feeding according to the following molar charge ratio, ligand 2: ligand 1: NH (NH)₄ReO₄: anhydrous stannous chloride 5:1:1:4, these starting materials were dissolved in water, adjusted to pH 6 using gentisic acid, boiled for 30 minutes, extracted with dichloromethane, and the dichloromethane was spun off to give the ester, i.e., the following compound i-1-Re, which was purified by HPLC (water: methanol 8:2), see fig. 2 and 3, ESI 993.48.

The resulting compound i-1-Re was dissolved in methanol and reacted with 0.3N NaOH at 70 ℃ for 10min, cooled and acidified to pH 2 with 0.1N HCl, extracted with dichloromethane and purified by HPLC (water: methanol 8:2) to give the product acid (R ═ H), i.e. the final product i-2-Re, see fig. 4-5, ESI ═ 965.50.

Target product I-1-^99mTc synthesis: ligand 1mg (ligand 2 to ligand 1 molar ratio 5: 1) was added, dissolved in 700. mu.L of physiological saline in a 10ml vial, and then 200. mu.L of LCB (citrate buffer, 0.1m, pH 6.6) and 20. mu.L of SnCl were added₂·2H₂O solution (1 mg SnCl)₂·2H₂O/1 ml) was added to the vial. Adding 100 μ L of water^99mTcO₄ ^-After the brine of (370) -3700Mbq), the solution was heated at 100 ℃ for 30 minutes. Extracting with dichloromethane, purging with nitrogen to remove dichloromethane to obtain corresponding ester, i.e. target product I-1-^99mTc, peak position by HPLC (water: methanol ═ 8:2) (see liquid phase fig. 6) consistent with the corresponding ester of i-1-Re, indicating the corresponding structure.

Then the target product I-1-^99mTc is dissolved in 200 mu L methanol, treated with 50 mu L0.3N NaOH and reacted at 70 ℃ for 10min, cooled, acidified with 150 mu L0.1N HCl, and extracted with dichloromethane to obtain the target product I-2-^99mTc, purified by HPLC (water: methanol ═ 8:2), and the spectrum is shown in fig. 7, which is substantially identical to the liquid phase spectrum of i-2-Re, indicating the same structure.

Target product I-1-^99mBiodistribution of Tc: female KM mice (22-25g, n ═ 5) were fasted for 12h before the experiment, and the purified radioactive complex I-1-^99mTc was prepared as a 100. mu. Ci/mL solution in physiological saline (10% ethanol). 100. mu.L of the above solution was diluted to 10mL as% ID. Injecting 100 μ L above solution via tail vein, cutting neck at five time points of 5,15,30, 60min and 120min, collecting blood, brain, heart, liver, spleen, lung, kidney, muscle, bone and tail, weighing, and making into capsuleThe counts were measured and the distribution of counts (unit:% ID/g) for each tissue and organ was calculated

TABLE 2M ═^99m(ii) a biodistribution of Tc, m 1, n 5 (% ID/g, n 5 mean. + -. SD)

From the above results, it was found that the compound of the present invention is very excellent in effect.

In this example, due to the labelled technetium^99mTc has radioactivity and cannot be directly measured, so that a rhenium Re-labeled target product is generally synthesized, and a mass spectrum, a liquid phase and the like of the target product are measured, and if the product has no problem, technetium is prepared by the same method^99mMeasuring the liquid phase of Tc-labeled target product, comparing the liquid phase spectrum with that of Re-labeled target product, and determining technetium if the liquid phase spectrum is basically consistent with that of Re-labeled target product^99mStructure of Tc labeled target product. Other embodiments of the invention are also identified by this method.

Example 4

Wherein m is 1, n is 3, R, R1, R2, R3, R4 and R5 are all CH₃The following compound I-2 was synthesized as an example.

1、

150mL of anhydrous methanol was charged into a 250mL round-bottom flask, 0.760mL (0.84g) of gamma-butyrolactone (99%) was added, 8.4mL of Et3N was measured, and the mixture was slowly added dropwise from a dropping funnel having a constant pressure and refluxed at 60 ℃ for 21 hours. The anhydrous methanol was distilled under reduced pressure, extracted with dichloromethane, and the dichloromethane was distilled under reduced pressure to obtain 1.06g of a product in 92% yield. The process can also be prepared by adopting the synthesis process of the ligand 1 in the example 2, namely, the compound 3 can be synthesized from the compound 2, and the compound 4 can be synthesized from the compound 3, namely the product.

2、

150mL of ethyl formate was added to a 250mL round bottom flask, 5.0g of 2-methylallyl amine (97%) was weighed into a constant pressure dropping funnel, refluxed in an oil bath at 60 ℃ for 3h, warmed to 70 ℃ and refluxed for 14h, and distilled under reduced pressure to give 6.49g of the product in 96% yield.

3、

20mL of anhydrous dichloromethane was weighed into a 100mL round-bottom flask, and the mixture was stirred at a raw material ratio of 1:1, 1.25N mercuric acetate is weighed and slowly added into the reaction solution, and the mixture is stirred for 14 hours at normal temperature. Filtering with filter paper, placing the filtrate into ice bath, and slowly adding 0.4N NaBH₄Stirring for 30min, and stirring for 4h at normal temperature. And filtering by using filter paper, pouring the filtrate into a fleroxacin funnel, performing suction filtration under reduced pressure, and distilling under reduced pressure to obtain a 0.73N product with the yield of 73 percent.

4、

Weigh 1.0N starting material into a round bottom flask with 20mL of anhydrous dichloromethane, add 2.45N of anhydrous Et₃N, weighing 1.1N anhydrous POCl₃The mixture was added to a constant pressure dropping funnel containing 10mL of anhydrous dichloromethane, and slowly added dropwise under ice bath conditions. Stirring for 15h at normal temperature. The reaction solution was added to 20% Na₂CO₃The solution was stirred for 3H, filtered, extracted 3 times with dichloromethane and distilled under reduced pressure to give the 0.82N product, ligand 1, in 82% yield, as shown in mass spectrum fig. 1, ESI (M + H: 199.9).

5、

Ligand 1(n ═ 3, R ═ H) and ligand 2 were dosed in a molar ratio of 1: 5, a total of about 4mg, dissolved in 700. mu.L of physiological saline in a 10ml vial, followed by 200. mu.L of CB (citrate buffer, 0.1m, pH 6.6) and 20. mu.L of SnCl₂·2H₂O solution (1 mg SnCl)₂·2H₂O/1 ml) was added to the vial. Adding 100 μ L of water^99mTcO₄ ^-After the brine of (370) -3700Mbq), the solution was heated at 100 ℃ for 30 minutes. Dichloromethane extraction, nitrogen purge to remove dichloromethane, then dissolved in 200 μ L methanol and treated with 50 μ L0.3N NaOH for 10min at 70C, cooled and acidified with 150 μ L0.1N HCl, extracted with dichloromethane and purified by HPLC (water: methanol 8:2) to give the final product i-2.

6. Biodistribution of the end product I-2

Female KM mice (22-25g, n ═ 5) were fasted for 12h before the experiment, and the purified radio-complex I-2 was prepared as a 100. mu. Ci/mL solution in physiological saline (containing 10% ethanol). 100. mu.L of the above solution was diluted to 10mL as% ID. 100 μ L of the above solution was injected via tail vein, and at five time points of 5,15,30 and 60min, 120min, respectively, neck was cut off and sacrificed, blood, brain, heart, liver, spleen, lung, kidney, muscle, bone and tail were collected, their masses were weighed and counted, and the distribution of counts (unit:% ID/g) of each tissue and organ was calculated

TABLE 1 biodistribution (% ID/g, n. RTM. 5, mean. + -. SD) of the final product I-2

The above results show that the compounds of the present invention are excellent in effect.

In this example, the target product identification method is identical to example 3, and the final product spectrum and the synthesis of the corresponding rhenium marker are omitted.

Example 5

By M ═^99mTc, m 6, n 5, R6 is ethyl or H, as examples to test the biodistribution of the following compounds I-3 and I-4The situation is.

The synthetic route is as follows:

the synthesis of ligand 1 is in accordance with example 3 and is not described in detail here.

Synthesis of Compounds I-3-Re: ligand 1, NH₄ReO₄And anhydrous stannous chloride dissolved in water (fed according to the following molar feed ratio, ligand 1: NH)₄ReO₄: anhydrous stannous chloride 6:1:4 in water), pH 6 adjusted using gentisic acid, boiling for 30 minutes, extraction with dichloromethane, and removal of dichloromethane to give the ester (R is aliphatic), i.e. i-3-Re, see figures 8 and 18, ESI 1633.76.

The compound i-3-Re was then dissolved in methanol and reacted with 0.3N NaOH at 70 ℃ for 10min, cooled and acidified to pH 2 with 0.1N HCl, extracted with dichloromethane and purified by HPLC (water: methanol 8:2) to give the acid (R ═ H), i.e. the compound i-4-Re, see fig. 9-10, ESI ═ 1465.64.

Target product I-3-^99mTc synthesis: ligand (n ═ 5, R ═ CH)₂CH₃)1mg, dissolved in 700. mu.L of physiological saline in a 10ml vial, followed by 200. mu.L of LCB (citrate buffer, 0.1m, pH 6.6) and 20. mu.L of SnCl₂2H2O solution (1 mg SnCl)₂·2H₂O/1 ml) was added to the vial. Adding 100 μ L of water^99mTcO₄ ^-After the brine of (370) -3700Mbq), the solution was heated at 100 ℃ for 30 minutes. Extracting with dichloromethane, purging with nitrogen to remove dichloromethane to obtain corresponding ester, i.e. target product I-3-^99mTc, liquid phase diagram thereof seeFIG. 19, which shows a peak time substantially identical to that of I-3-Re.

Target product I-4-^99mTc synthesis: then dissolving the product obtained above in 200 μ L methanol, treating with 0.3N NaOH 50 μ L at 70C for 10min, cooling, acidifying with 0.1N HCl 150 μ L, extracting with dichloromethane to obtain final product I-4-^99mTc, purified by HPLC (water: methanol ═ 8:2), and the liquid phase spectrum is shown in fig. 11, the peak time substantially coincides with that of i-4-Re.

End product I-4-^99mBiodistribution of Tc: female KM mice (22-25g, n ═ 5) were fasted for 12h before the experiment, and the purified radioactive complex I-4-^99mTc was prepared as a 100. mu. Ci/mL solution in physiological saline (10% ethanol). 100. mu.L of the above solution was diluted to 10mL as% ID. 100 μ L of the above solution was injected via tail vein, and at five time points of 5,15,30 and 60min, 120min, respectively, neck was cut off and sacrificed, blood, brain, heart, liver, spleen, lung, kidney, muscle, bone and tail were collected, their masses were weighed and counted, and the distribution of counts (unit:% ID/g) of each tissue and organ was calculated.

TABLE 3M ═^99m(iv) a biodistribution of Tc, m 6, n 5 (% ID/g, n 5 mean. + -. SD)

The above results show that the overall effect is better.

Example 6

Synthesis of the target products I-5 and I-6 in the general formula III.

The synthesis process comprises the following steps:

5mmol of Compound 1 are dissolved in 20ml of methanol in a round-bottomed flask, 20ml of ethyl formate and 750. mu.l of triethylamine are then added, refluxed for 36 hours, the solvent is removed by spinning off, the residue is dissolved in dichloromethane and washed three times with water, and the oily phase is spun off as 46% colorless oil 2.

Compound 2(3.0mol) was dissolved in 15ml of methylene chloride, and 15.0mol of triethylamine was added. 3.3mmol of POCl was added dropwise at 0 deg.C₃At 30min, room temperature for 3h and then saturated NaHCO₃Quenching with anhydrous MgSO₄Drying, filtering and concentrating, and passing through a flash silica gel column (mobile phase is dichloromethane) to obtain a compound 3, namely the ligand 3, wherein the yield is 89%, and mass spectra are measured and comprise the following mass spectrum data: ESI (M + H) ═ 170.3.

Ligand 2

See example 3 for a synthetic method of (1).

Synthesis of the final product I-5-Re: feeding according to the following molar charge ratio, and feeding a ligand 2: ligand 3: NH (NH)₄ReO₄: anhydrous stannous chloride 5:1:1:4 was dissolved in water, the solution was adjusted to pH 5 using gentisic acid and boiled for 30 minutes, extracted with dichloromethane and the dichloromethane was spun off to give the corresponding ester complex i-5-Re, which was purified by HPLC (water: methanol 8:2) as shown in fig. 12-13, ESI 921.78.

Synthesis of the final product I-6-Re: i-5-Re was dissolved in methanol, reacted with 0.3N NaOH at 70 ℃ for 10min, cooled and acidified to pH 2 with 0.1N HCl, and extracted with dichloromethane to give the corresponding acid i-6-Re, which was purified by HPLC (water: methanol 8:2), as shown in fig. 14-15, ESI 907.49.

Target product I-5-^99mTc synthesis: ligand 2 and ligand 3 were dosed in a molar ratio of 5:1, approximately 4mg total, dissolved in 700 μ L physiological saline in a 10ml vial, followed by 200 μ L CB (citrate buffer, 0.1m, pH 6.6) and 20 μ L SnCl₂·2H₂O solution (1 mg SnCl)₂·2H₂O/1 ml) was added to the vial. Adding 100 μ L of water^99mTcO₄ ^-After the brine of (370-3700Mbq), the solution was heated at 100 ℃ for 30 minutes; extracting with dichloromethane, purging with nitrogen to remove dichloromethane to obtain corresponding ester, i.e. target product I-5-^99mTc, corresponding structure is shown by the consistent time of peak position by HPLC (water: methanol 8:2) (see liquid phase FIG. 16) with ester I-5-Re.

Target product I-6-^99mTc synthesis: the I-5-^99mTc is dissolved in 200 mul methanol, treated by 50 mul of 0.3N NaOH and reacted for 10min at 70 ℃, cooled and acidified by 150 mul of 0.1N HCl, and then extracted by dichloromethane to obtain the target product I-6-^99mTc, purified by HPLC (water: methanol ═ 8:2), as shown in fig. 17, which is essentially identical to the corresponding liquid phase pattern for i-6-Re, indicating the same structure.

The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A labeled isocyano group-containing fatty acid derivative, characterized in that: it is the following compound:

wherein M is labelled technetium^99mTc or Re, R, R₁、R₂、R₃、R₄And R₅Selected from H or aliphatic hydrocarbon with 1-4C, and n is a positive integer of 1-10.

2. Use of the labeled isocyano group-containing fatty acid derivative according to claim 1 in the preparation of a myocardial imaging agent.