A labeled glucosamine derivative, its preparation method, its precursor and its application

Abstract

The present invention provides a labeled glucosamine derivative, a preparation method thereof, a precursor thereof and an application thereof, belonging to the technical fields of medicine and chemistry. The general formula of the glucosamine derivative is shown in Formula I. For the above-mentioned derivatives protected by the present invention, the overall spatial volume and the overall molecular weight are greatly reduced, which can enhance the potential of such derivatives to penetrate the cell membrane phospholipid layer and the potential to be successfully transported by glucose transporters. Moreover, its residence time in tumors is long and the absolute uptake value is increased, so it can be used as a new type of tumor metabolic imaging agent; in addition, the present application has developed a brand-new precursor. Compared with the prior art, the synthesis of Formula I is simple, the synthesis difficulty of the precursor is greatly reduced, the chemical stability of the precursor is significantly enhanced, which is conducive to the realization of the kit of the technetium-labeled complex, and has important clinical application prospects.

Description

Technical Field

[0001] The present invention relates to the technical fields of medicine and chemistry, and particularly relates to a labeled glucosamide derivative, a preparation method thereof, a precursor thereof, and an application thereof. Background Art

[0002] Currently, malignant tumors have become the number one killer threatening human health and life. The annual incidence and mortality of tumors are still on the rise. Early diagnosis and treatment of tumors are the most effective measures to reduce tumor mortality. Traditional non-invasive tumor detection imaging methods such as X-ray, CT, MRI, etc. mainly show the morphology and structure of organs in the pathological state, while nuclear medicine molecular imaging technologies such as positron emission tomography (PET) and single photon emission computed tomography (SPECT) can reflect the physiological, pathological, metabolic, and functional changes of tumors at the molecular level. Currently, with the organic integration of imaging technologies such as PET / CT, PET / MR, and SPECT / CT playing an increasingly important role in the early diagnosis and personalized treatment of tumors, to meet the wide application and rapid development needs of nuclear medicine molecular imaging technologies in clinical diagnosis and treatment, the research on radioactive tumor molecular probes on which this technology depends has become particularly urgent and important.

[0003] Currently, the most widely used tumor imaging agent in clinical practice is 18 F-fluorodeoxyglucose ( 18 F-FDG), which has good imaging effects, but 18 the radionuclide 99m F is relatively expensive. Therefore, to a certain extent, it limits its wide application, especially in economically underdeveloped countries and regions where its application is not yet popular. In addition, 99m Tc is the most widely used radionuclide in nuclear medicine imaging applications. It has ideal nuclear physical properties (half-life of about 6 h), low cost, easy access, and can form various coordination structures. Currently, there are various labeled methods that have been made into kits, which are convenient for clinical application and drug distribution. In 2019, the market value of only 99m the

[0004] Isocyanide is a class of organic compounds with the general formula R-N + ≡C - . Research shows that the carbon atom in isocyanide can coordinate with 6 99mTc to form a +1-valent + complex 99mTechnetium-methoxyisobutylisonitrile( 99m Tc-MIBI) has been widely used clinically as a myocardial perfusion imaging agent, and it has also been found clinically that this imaging agent also has a certain tumor affinity. The isonitrile molecule connects 99m Tc and the sugar molecule in the tumor molecular probe of the present invention, acting as a bifunctional linker, integrating the tumor-affine sugar metabolism function with 99m Tc tracer function. Based on the above background, someone has previously developed and explored a tumor molecular probe of 99m Tc-labeled glucose, converting glucosamine into a glucose derivative containing isonitrile (abbreviated as CNDG), which has relatively high tumor / blood, tumor / muscle, tumor / liver, and tumor / lung values. However, since each ligand in its complex contains a glucose-like structure with a relatively large space and molecular weight (the molecular weight is close to 2000, which is very large), and its water solubility is too strong (most logP values are lower than -3.50), it may be difficult for such complexes to penetrate the cell membrane phospholipid layer and be transported by glucose transporters, resulting in relatively weak absolute tumor uptake values. In addition, in the prior synthesis of 99m Tc-labeled glucose-like molecules, the precursors used (such as: Patent 201710451094.8) have the defect of instability during long-term storage, and the separation difficulty is relatively large. Therefore, it is of extremely important practical significance to design a glucose amide derivative containing a small molecular weight isocyano ligand. Summary of the Invention

[0005] The object of the present invention is to provide a labeled glucose amide derivative, its preparation method, its precursor and its application. In this derivative, the inventors have studied a series of small molecular weight isocyano ligands to mix with it. Compared with the original isocyano glucose amide tumor imaging agent molecule (pure coordination compound of technetium or rhenium), the overall molecular weight of the complex is greatly reduced, enhancing the potential of this type of derivative to penetrate the cell membrane phospholipid layer and be successfully transported by glucose transporters. In addition, aiming at the defect of instability during long-term storage of the original isocyano monomeric glucose amide labeling precursor, the inventors have developed a new precursor compound, which has greatly reduced separation difficulty and greatly improved chemical stability compared with the previous labeling precursor; and the inventors have also given for the first time a simple method for synthesizing the above precursor and derivative. The present invention solves the deficiencies existing in the prior art.

[0006] The technical solution of the present invention is realized as follows:

[0007] One inventive point of the present invention is to provide a labeled glucose amide derivative, whose general formula is shown in Formula I:

[0008]

[0009] Among them, M is 99m Tc or Re;

[0010] R is H, an aliphatic chain or an alicyclic group;

[0011] A 1 -A 4 are independently H, an aliphatic chain or an alicyclic group;

[0012] J is at least one of, where R 1 , R 2 , R 3 and R 4 are independently H, an aliphatic chain or an alicyclic group;

[0013] X is O or S;

[0014] a is an integer from 1 to 7, b is an integer from 1 to 12, and z is an integer from 1 to 6.

[0015] As a further improvement of the present invention, J is at least one of.

[0016] As a further improvement of the present invention, X is O.

[0017] As a further improvement of the present invention, R, R 1 , R 2 , R 3 and R 4 are independently H or an aliphatic chain; more preferably, the aliphatic chain includes aliphatic hydrocarbons.

[0018] As a further improvement of the present invention, A 1 , A 2 , A 3 , A 4 are independently H or an aliphatic chain; more preferably, the aliphatic chain includes aliphatic hydrocarbons.

[0019] As a further improvement of the present invention, the above-mentioned aliphatic chain is an aliphatic hydrocarbon having 1 to 15 carbon atoms.

[0020] As a further improvement of the present invention, R, R 1 , R 2 , R 3 and R 4 and A 1 -A 4 are independently H atoms or aliphatic hydrocarbons having 1 to 5 carbon atoms, that is, independently -H, -CH 3 , -CH 2 CH 3 , -CH 2 CH2 CH 3 、-CH(CH 3 )CH 3 、-CH 2 CH 2 CH 2 CH 3 、-CH(CH 3 )CH 2 CH 3 、-CH 2 CH(CH 3 )CH 3 、-C(CH 3 ) 3 、-CH 2 CH 2 CH 2 CH 2 CH 3 、-CH(CH 3 )CH 2 CH 2 CH 3 、-CH 2 CH(CH 3 )CH 2 CH 3 、-CH 2 CH(CH 3 )CH 2 CH 3 、-CH 2 CH 2 CH(CH 3 )CH 3 、-C(CH 3 ) 2 CH 2 CH 3 、-CH 2 C(CH 3 ) 2 CH 3 or -CH(CH 3 )CH(CH 3 )CH 3 .

[0021] As a further improvement of the present invention, z is an integer between 1 and 5, such as 1, 2, 3, 4, or 5.

[0022] As a further improvement of the present invention, a is an integer of 1-5, such as 1, 2, 3, 4, 5; b is an integer of 2-10, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10.

[0023] As a further improvement of the present invention, M is 99mTc or Re, z is an integer from 1 to 5, a is an integer from 1 to 5, b is an integer from 2 to 10, R, R 1 , R 2 , R 3 and R 4 are independently -H, -CH 3 or -CH 2 CH 3 , A 1 -A 4 are both -H, when X is an O atom, then General Formula I includes the following compounds: ① When J is , Formula I is the following General Formula II: ② When J is , Formula I is the following General Formula III: ③ When J is , Formula I is the following General Formula IV:

[0024] As a further improvement of the present invention, when z is an integer from 1 to 5, a is an integer from 1 to 5, b is an integer from 2 to 10, R is -H, -CH 3 or -CH 2 CH 3 , A 1 -A 4 are both -H, when X is an O atom, when J is , General Formula II includes but is not limited to the following compounds:

[0025]

[0026] When z is an integer from 1 to 5, a is an integer from 1 to 5, b is an integer from 2 to 10, A 1 -A 4 are both -H, when X is an O atom, when J is R, R 1 are independently -H, -CH 3 or -CH 2 CH 3 , General Formula III includes but is not limited to the following compounds:

[0027] When z is an integer from 1 to 5, a is an integer from 1 to 5, b is an integer from 2 to 10, A 1 -A 4 are both -H, when X is an O atom, when J is R, R 2 are independently -H, -CH 3 or -CH 2 CH 3 , General Formula IV includes but is not limited to the following compounds:

[0028] As a further improvement of the present invention, the structural general formula of the labeled precursor of the glucosamine derivative in the right part of the general formula Ⅰ is a metal salt, that is The structural general formula of the labeled precursor in the left part of the general formula Ⅰ is also a metal salt, that is

[0029] wherein, Q 1 and Q 2 are independently metal cations, and the metal cation is preferably a copper ion, a cuprous ion, a calcium ion, a potassium ion, a sodium ion, a magnesium ion, an aluminum ion, and more preferably a copper ion or a cuprous ion;

[0030] E 1 and E 2 are independently anions, and the anion is preferably a tetrafluoroborate ion (BF 4 - ), a hexafluorophosphate ion (PF 6 - ), a trifluoroacetate ion (CF 3 COO - ), a perchlorate ion (ClO 4 - ), a sulfate ion (SO 4 2- ), a fluoride ion, a chloride ion, a bromide ion, an iodide ion, and more preferably a tetrafluoroborate ion (BF 4 - );

[0031] A 1 ~A 4 , X, J, a and b are all the same as those defined in any of the above paragraphs;

[0032] n and m vary according to different cations and anions, as long as they conform to the coordination rule. For the above-mentioned cations and anions, n and m are usually values between 2 and 6, such as 2, 3, 4, 5, 6. For example, when Q is a cuprous ion and E is a tetrafluoroborate ion, n is 4;

[0033] The metal salts of the left and right parts can directly synthesize the general formula Ⅰ through a one-step labeling method.

[0034] Another inventive point of the present invention is to provide a precursor compound of a glucosamine derivative labeled as described in any of the above paragraphs, and the precursor compound is and / or

[0035]

[0036] wherein, Q 1 and Q 2Independently a metal cation, which is preferably a copper ion, cuprous ion, calcium ion, potassium ion, sodium ion, magnesium ion, aluminum ion, more preferably a copper ion or cuprous ion;

[0037] E 1 and E 2 Independently an anion, which is preferably a tetrafluoroborate ion (BF 4 - ), hexafluorophosphate ion (PF 6 - ), trifluoroacetate ion (CF 3 COO - ), perchlorate ion (ClO 4 - ), sulfate ion (SO 4 2- ), fluoride ion, chloride ion, bromide ion, iodide ion, more preferably a tetrafluoroborate ion (BF 4 - );

[0038] A 1 ~A 4 、X, J, a and b are all the same as those defined in any of the above paragraphs;

[0039] n and m vary according to different cations and anions, as long as they conform to the coordination rules. For the above-mentioned cations and anions, n and m are usually values between 2 and 6, such as 2, 3, 4, 5, 6. For example, when Q is a cuprous ion and E is a tetrafluoroborate ion, n is 4;

[0040] The metal salts on both left and right sides can directly synthesize the general formula I by a one-step labeling method.

[0041] Another inventive point of the present invention is to provide a preparation method of a labeled glucosamine derivative as described in any of the above paragraphs. The method is as follows: First, synthesize the precursors and Then, the or react with to synthesize the derivative shown in the general formula I by a one-step labeling method.

[0042] For example, the precursor compound of the labeled glucosamine derivative described above is a cuprous salt The precursor compound of the labeled glucosamine derivative is a cuprous salt Then, the two cuprous salts can be directly synthesized into the general formula I through a one-step labeling method. Among them, the precursor compounds of the above two labeled glucosamine derivatives can be directly precipitated from the reaction, separated and purified by suction filtration, and their chemical properties are very stable. Previously, derivatives with a similar isocyanoglucosamine structure used isocyanoglucosamine monomeric labeling precursors for labeling preparation, which had relatively poor stability and complex separation. Therefore, the method of the present invention overcomes the instability of the previous isocyanoglucosamine monomeric labeling precursors and the complex silica gel column chromatography separation steps required for the isocyanoglucosamine monomeric labeling precursors, greatly simplifying the separation process.

[0043] Another inventive point of the present invention is to provide an application of the above-labeled glucosamine derivatives in the preparation of tumor imaging agents.

[0044] The last inventive point of the present invention is to provide a tumor imaging agent, which comprises the compound shown in formula I described in any of the above paragraphs.

[0045] The present invention has the following beneficial effects:

[0046] The present invention synthesizes a brand-new structure, which creatively adds a small molecular weight coordination group (mixedly coordinated with technetium or rhenium together with isocyanoglucosamine) to the structure of isocyanoglucosamine compounds. Thus, compared with the original isocyanoglucosamine tumor imaging agent molecules (pure coordination compounds of technetium or rhenium), while maintaining water solubility or slightly increasing water solubility, the spatial volume and overall molecular weight of the complex are greatly reduced, enabling the potential of such derivatives to penetrate the cell membrane phospholipid layer and the potential to be successfully transported by glucose transporters to be enhanced.

[0047] In order to achieve better application of the above derivatives, the inventor of the present invention also made an innovative exploration of the structure of the precursor. The chemical stability of the precursor is greatly improved, and it can be stored stably for a long time, which is beneficial to the realization of later commercial applications. Moreover, it is easy to synthesize glucosamine derivatives from the precursor with good effects.

[0048] In addition, the present inventor has synthesized for the first time isocyanoglucosamide salt-labeled precursors for the preparation of isocyanoglucosamide-based tumor imaging agents. Compared with the original isocyanoglucosamide monomer-labeled precursors (patent application number: 201710451094.8), the products of this type of labeled precursors can directly precipitate from the reaction mixture, and pure copper isocyanoglucosamide salt-labeled precursors can be obtained through simple filtration and washing (the original highly polar isocyanoglucosamide monomers require complex silica gel column chromatography purification operations). This not only greatly reduces the separation difficulty but also significantly improves the chemical stability, facilitating long-term storage and the realization of the kit form of future technetium-labeled drugs; the vast majority of other small-molecular-weight copper isocyanide salt-labeled precursors can also precipitate naturally from the reaction system, thus greatly reducing the separation difficulty of these labeled precursors.

[0049] The labeled glucosamide derivatives claimed in this application are a completely new structural type. These derivatives (imaging agents) have a long retention time in tumors and an increased absolute uptake value. This type of derivative is conducive to the realization of kit form, has important clinical application prospects, and has great potential to become a new type of tumor metabolic imaging agent. Detailed implementation manners

[0050] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Apparently, the described embodiments are only a part of the embodiments of the present invention, rather than all of them. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.

[0051] The specific synthesis processes of the compounds in II, III, and IV separately defined in the present invention will be introduced below. Since it is impossible to list all the protected compounds in the synthesis process, those not listed can refer to the following methods, which can be completely achieved by those skilled in the art. And due to space limitations, only a few structures in this application are used to illustrate the effects, but the structures not listed also have similar effects.

[0052] Example 1

[0053] Select the precursor compounds of the glucosamide derivatives labeled with general formulas I-IV - copper isocyanoglucosamide salt-labeled precursors 19-21 (listing the right parts in general formulas I-IV and b in the general formula being 3, 5, and 7 respectively, Q 1 being a copper ion, E 1 being a tetrafluoroborate ion, and n being 4), as follows:

[0054] (1) The synthesis route is as follows:

[0055]

[0056] (2) The synthesis process is as follows:

[0057] Synthesis of Intermediate 4: At 0 °C, formic acid (22.3187 g) was added to acetic anhydride (49.5006 g). Then, the mixture was stirred at 60 °C for 2 h to obtain acetic-methyl anhydride. The newly prepared acetic-methyl anhydride was added to a DMF solution of starting material 1 (20.0000 g), and the mixture was stirred overnight at room temperature. The mixture was concentrated under vacuum, and an appropriate amount of a mixed solution of ether / ethyl acetate / acetonitrile (the ratio between ether / ethyl acetate / acetonitrile is relatively free, for example, the volume ratio can be 1:(0.1 - 10):(0.1 - 10), here it is 6:1:1) was added to the obtained concentrated solution. Intermediate 4 (white solid, 14.5035 g) was obtained by filtration and was directly used in the next reaction without further purification.

[0058] Synthesis of Intermediate 5: For the specific operation, refer to "Synthesis of Intermediate 4". After acetic anhydride (38.9122 g) and formic acid (17.5446 g) reacted to obtain acetic-methyl anhydride, it was then reacted with starting material 2 (20.0000 g), and Intermediate 5 (white solid, 19.8102 g) was obtained by filtration and was directly used in the next reaction without further purification.

[0059] Synthesis of Intermediate 6: For the specific operation, refer to "Synthesis of Intermediate 4". After acetic anhydride (40.3019 g) and formic acid (18.1712 g) reacted to obtain acetic-methyl anhydride, it was then reacted with starting material 3 (25.1436 g). A white solid precipitated from the mixture. After the white solid was filtered under reduced pressure and washed with ethyl acetate, pure Intermediate 6 (white solid, 20.1269 g) was obtained. Its characterization data: 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.94 (s, 1H), 7.98 (s, 1H), 7.88 - 7.97 (m, 1H), 3.06 (q, J = 6.6 Hz, 2H), 2.19 (t, J = 7.3 Hz, 2H), 1.44 - 1.55 (m, 2H), 1.33 - 1.43 (m, 2H), 1.25 (s, 6H).

[0060] Synthesis of Intermediate 8: Compound 7 (15.2101 g) was added to the DMF solution of Intermediate 4 (10.0000 g). Then, the mixture was stirred at 0 °C for 0.5 h, after which a DMF solution of N,N'-dicyclohexylcarbodiimide (DCC, 22.1707 g) was added thereto, and the mixture was stirred overnight at room temperature. The mixture was filtered under reduced pressure, and the filtrate was concentrated in vacuo to obtain crude Intermediate 8, which was used directly in the next reaction without further purification.

[0061] Synthesis of Intermediate 9: For the specific operation, refer to "Synthesis of Intermediate 8". Intermediate 5 (10.0000 g) was reacted with 7 (12.5298 g) and DCC (18.2635 g) to obtain crude Intermediate 9, which was used directly in the next reaction without further purification.

[0062] Synthesis of Intermediate 10: For the specific operation, refer to "Synthesis of Intermediate 8". Intermediate 5 (22.8937 g) was reacted with 7 (24.3866 g) and DCC (35.5460 g) to obtain crude Intermediate 10, which was used directly in the next reaction without further purification. The characterization data thereof are as follows: 1 H NMR (400 MHz, CDCl 3 ): δ 8.16 (s, 1H), 7.03 (tt, J = 9.9, 7.1 Hz, 1H), 5.90 (brs, 1H), 3.20 - 3.23 (m, 2H), 2.67 (t, J = 7.3 Hz, 2H), 1.75 - 1.82 (m, 2H), 1.53 - 1.56 (m, 2H), 1.39 - 1.44 (m, 6H).

[0063] Synthesis of Intermediate 11: Triethylamine (5.6913 g) was added to the CH 2 Cl 2 solution of Intermediate 8. Then, trichloromethyl chloroformate (2.2515 g) in CH 2 Cl 2 solution was added to the mixture at -30 °C. Then, the mixture was stirred at room temperature for 30 min and quenched with an aqueous solution of 20% potassium carbonate. The mixture was extracted with dichloromethane, filtered under reduced pressure, and the filtrate was concentrated in vacuo. The concentrate was separated by silica gel column chromatography to obtain relatively pure Intermediate 11 (0.6367 g). The characterization data thereof are as follows: 1 H NMR (400 MHz, CDCl 3 ) δ 7.03 (tt, J = 9.9, 7.1 Hz, 1H), 3.54 - 3.64 (m, 2H), 2.91 (t, J = 7.1 Hz, 2H), 2.10 - 2.23 (m, 2H).

[0064] Synthesis of Intermediate 12: For the specific operation, refer to "Synthesis of Intermediate 11". React Intermediate 9 with triethylamine (7.8667 g) and triphosgene (19.8845 g) to obtain relatively pure Intermediate 12 (2.4300 g), which is directly used for the next reaction.

[0065] Synthesis of Intermediate 13: For the specific operation, refer to "Synthesis of Intermediate 11". React Intermediate 10 with triethylamine (12.0388 g) and triphosgene (4.7627 g) to obtain Intermediate 13 (4.2301 g). Its characterization data are as follows: 1 H NMR (400 MHz, CDCl 3 ) δ 7.00 (dqd, J = 9.6, 7.2, 3.5 Hz, 1H), 3.40 (qd, J = 4.4, 1.8 Hz, 2H), 2.68 (td, J = 7.4, 2.2 Hz, 2H), 1.81 (p, J = 6.6 Hz, 2H), 1.71 (t, J = 6.2 Hz, 2H), 1.54 - 1.35 (m, 6H); 13 C NMR (101 MHz, CDCl 3 ) δ 169.42, 155.89, 155.83, 155.77, 147.40, 147.36, 147.28, 147.24, 147.16, 147.12, 144.93, 144.89, 144.81, 144.77, 144.65, 141.95, 141.78, 139.47, 139.32, 129.86, 129.72, 129.58, 103.34, 103.12, 102.89, 77.34, 77.03, 76.71, 41.55, 41.48, 41.42, 33.29, 28.95, 28.54, 28.26, 26.10, 24.59.

[0066] Synthesis of Copper(I) Isocyanotetrafluorophenolate Intermediate 15: Stir Intermediate 11 (636.7 mg) and copper(I) tetrafluoroborate tetrakis(acetonitrile) (191.7 mg) in dichloromethane at room temperature for 30 - 60 min. Filter by suction, concentrate the filtrate under vacuum, and separate the concentrate by silica gel column chromatography to obtain copper(I) isocyanotetrafluorophenolate Intermediate 15 (501.5 mg). Its characterization data are as follows: 1 HNMR (400 MHz, CDCl 3 ) δ 7.02 (tt, J = 9.9, 7.0 Hz, 4H), 3.81 (t, J = 6.7 Hz, 8H), 2.93 (t, J = 7.1 Hz, 8H), 2.19 - 2.34 (m, 8H).

[0067] Synthesis of isocyanotetrafluorophenolate copper salt intermediate 16: For the specific operation, refer to "Synthesis of isocyanotetrafluorophenolate copper salt intermediate 15". Isocyanotetrafluorophenolate copper salt intermediate 16 (2.0310 g) was obtained by reacting intermediate 12 (2.4300 g) with copper(I) tetra(acetonitrile) tetrafluoroborate (660.7 mg). Its characterization data: 1 HNMR(400MHz,CDCl 3 )δ7.01(tt,J=10.0,7.0Hz,4H),3.62(t,J=6.6Hz,8H),2.73(t,J=7.2Hz,8H),1.84(p,J=7.2Hz,16H),1.52-1.63(m,8H).

[0068] Synthesis of isocyanotetrafluorophenolate copper salt intermediate 17: For the specific operation, refer to "Synthesis of isocyanotetrafluorophenolate copper salt intermediate 15". Isocyanotetrafluorophenolate copper salt intermediate 17 (373.2 mg) was obtained by reacting isocyanotetrafluorophenolate intermediate 13 (1.1608 g) with copper(I) tetra(acetonitrile) tetrafluoroborate (288.7 mg). Its characterization data: 1 H NMR(400MHz,CDCl 3 )δ7.00(tt,J=9.9,7.0Hz,4H),3.60(t,J=6.9Hz,8H),2.68(t,J=7.2Hz,8H),1.78(p,J=7.3,6.7Hz,16H),1.37-1.51(m,24H).

[0069] Synthesis of isocyanoglucosamide copper salt labeling precursor 19: D-glucosamine hydrochloride (165.2 mg) 18 and sodium hydroxide (40.0 mg) were stirred in methanol at room temperature for 30 min, then isocyanotetrafluorophenolate copper salt intermediate 15 (256.0 mg) was added, and the mixture was stirred overnight at room temperature. The naturally precipitated white solid was filtered by suction and washed with a small amount of methanol. The filter cake was dried under vacuum to remove the residual solvent, obtaining isocyanoglucosamide copper salt labeling precursor 19 (120.5 mg). Its characterization data: 1 H NMR(400MHz,DMSO-d 6) δ 7.72 - 8.07 (m, 4H), 6.50 (dd, J = 36.1, 5.3 Hz, 4H), 4.83 - 5.25 (m, 8H), 4.48 - 4.78 (m, 4H), 4.33 - 4.52 (m, 4H), 3.68 - 3.81 (m, 8H), 3.56 - 3.64 (m, 8H), 3.40 - 3.56 (m, 8H), 3.04 - 3.14 (m, 4H), 2.11 - 2.41 (m, 8H), 1.88 (p, J = 7.0 Hz, 8H), 0.99 - 1.37 (m, 4H); 13 C NMR (101 MHz, DMSO - d 6 ) δ 171.42, 91.03, 72.55, 71.56, 71.24, 70.93, 61.55, 55.38, 54.78, 32.02, 25.03; 19 F NMR (376 MHz, DMSO - d 6 ) δ - 148.24; HRMS (High Resolution): calcd. for C 22 H 36 CuN 4 O 12 + 611.1620 (2M + Cu) + , found 611.1598 (2M + Cu) + 。Here, "M" refers to the exact mass of the isocyanocarboxylic acid monomer in the cuprous salt, "calcd." refers to the theoretically predicted value, and "found" refers to the measured value. The same applies hereinafter.

[0070] Synthesis of the cuprous salt of isocyanoglucosamide precursor 20: For the specific operation, refer to "Synthesis of the cuprous salt of isocyanoglucosamide precursor 19". D - glucosamine hydrochloride (228.4 mg) 18, sodium hydroxide (46.1 mg) and the cuprous salt intermediate of isocyanotetrafluorophenol ester 16 (386.3 mg) reacted to obtain the cuprous salt of isocyanoglucosamide precursor 20 (157.0 mg), and its characterization data: 1 H NMR (400 MHz, DMSO - d 6) δ 7.63 (dd, J = 39.1, 8.3 Hz, 4H), 6.45 (dd, J = 30.5, 5.3 Hz, 4H), 4.75 - 5.03 (m, 8H), 4.57 (dd, J = 30.9, 5.6 Hz, 4H), 4.34 - 4.49 (m, 4H), 3.67 - 3.75 (m, 8H), 3.56 - 3.63 (m, 8H), 3.39 - 3.55 (m, 8H), 3.05 - 3.15 (m, 4H), 1.98 - 2.24 (m, 8H), 1.66 (p, J = 7.0 Hz, 8H), 1.52 (p, J = 7.3 Hz, 8H), 1.33 (hept, J = 7.2, 6.7 Hz, 8H); 13 C NMR (101 MHz, DMSO - d 6 ) δ 172.60, 91.10, 74.79, 72.53, 71.66, 70.92, 61.58, 54.65, 43.07, 35.29, 28.15, 25.80, 24.77; 19 F NMR (376 MHz, DMSO - d 6 ) δ - 148.25; HRMS (high - resolution): calcd. for C 26 H 44 CuN 4 O 12 + 667.2246 (2M + Cu) + , found 667.2261 (2M + Cu) + , calcd. for C 13 H 23 N 2 O 6 + 303.1556 (M + H) + , found 303.1550 (M + H) + , calcd. for C 13 H 22 NaN 2 O 6 + 325.1376 (M + Na) + , found 325.1383 (M + Na) + , calcd. for C 39 H 66 CuN 6 O 18 + 969.3724 (3M + Cu) + , found 969.3732 (3M + Cu)+ , calcd. for C 52 H 88 CuN 8 O 24 + 1271.5202(4M + Cu) + , found 1271.5273(4M + Cu) + 。

[0071] Synthesis of copper isocyanoglucosamide precursor 21: For the specific operation, refer to "Synthesis of copper isocyanoglucosamide precursor 19". D - glucosamine hydrochloride (111.6 mg) 18, sodium hydroxide (22.6 mg) and copper isocyanotetrafluorophenolate intermediate 17 (213.9 mg) react to obtain copper isocyanoglucosamide precursor 20 (92.7 mg), and its characterization data are as follows: 1 H NMR (400 MHz, DMSO - d 6 ) δ 7.58 (dd, J = 58.2, 8.2 Hz, 4H), 6.42 (dd, J = 29.1, 5.4 Hz, 4H), 4.74 - 5.04 (m, 8H), 4.49 - 4.69 (m, 4H), 4.32 - 4.48 (m, 4H), 3.71 (t, J = 6.6 Hz, 8H), 3.56 - 3.64 (m, 8H), 3.44 - 3.54 (m, 8H), 3.17 (d, J = 4.4 Hz, 4H), 3.05 - 3.14 (m, 4H), 1.97 - 2.36 (m, 8H), 1.64 (p, J = 6.7 Hz, 8H), 1.48 (p, J = 7.2 Hz, 8H), 1.21 - 1.40 (m, 24H); 19 F NMR (376 MHz, DMSO - d 6 ) δ - 148.26; ESI - MS (low resolution): calcd. for C 15 H 26 CuN 2 O 6 + 393.1081(2M + Cu) + , found 393.1466(M + Cu) + , calcd. for C 30 H 52 CuN 4 O 12 + 723.2872(2M + Cu) + , found 723.3530(2M + Cu) + , calcd. for C15 H 26 NaN 2 O 6 + 353.1689(M + Na) + , found 353.2016(M + Na) + , calcd. for C 45 H 78 CuN 6 O 18 + 1053.4663(3M + Cu) + , found 1053.5664(3M + Cu) + 。

[0072] For this embodiment, it can be seen from the above experimental results that the present invention has synthesized for the first time the copper isocyanoglucosamide precursor of the right part in the isocyanoglucosamide tumor imaging agent. Compared with the original isocyanoglucosamide precursor existing in monomer form (Patent Application No.: 201710451094.8), the precursor of this isocyanocomplex can directly precipitate from the reaction mixture of the last step, and a pure copper isocyanoglucosamide precursor can be obtained only by simple filtration and washing (the highly polar isocyanoglucosamide monomer existing in monomer form originally requires complex silica gel column chromatography purification operation to be separated). Therefore, the method of the present invention not only greatly reduces the separation difficulty of the isocyanoglucosamide precursor, but also greatly improves its chemical stability, which is beneficial to long-term storage and the realization of the kit of technetium-labeled drugs in future clinical research. In addition, compared with the synthesis routes of other current researchers for the first and second steps in the synthesis steps of this precursor, the more polar reaction products in these two steps (such as intermediate 4-6 and intermediate 8-10 containing formamide groups in Example 1) can be directly used in the next step without purification, which greatly reduces the intermediate links of separation and purification, significantly saves the synthesis time of the precursor and reduces the synthesis workload; moreover, the third step reaction (the reaction for preparing isocyanotetrafluorophenol ester) does not require expensive Burgess reagent, nor does it need to react for 4 h, only 20-30 min of reaction is required, and after adding triphosgene, it can be completely restored to room temperature for stirring, and it does not need to be stirred at about 0 °C, which further saves the time required for precursor synthesis and reduces the synthesis cost; and, for the copper salt of cyanotetrafluorophenol ester, and then reacting the copper isocyanotetrafluorophenol ester with glucosamine, such a product is highly polar due to being tetracoordinated and can directly precipitate a part from the reaction system, thus greatly reducing the separation difficulty, and successfully preparing the copper isocyanoglucosamide precursor of the present invention. The synthesis route after the third step of the present invention has also been greatly improved compared with some existing technologies, that is, after preparing the isocyanotetrafluorophenol ester monomer with very low polarity, unlike the traditional idea, first reacting the isocyanotetrafluorophenol ester monomer with glucosamine to prepare the isocyanoglucosamide monomer with very high polarity (this will cause great separation difficulties in the later stage, and the excessive separation time may lead to the deterioration of the isocyanomonomer class), and then preparing the copper isocyanide salt, but creatively first preparing the intermediate of the isocyanotetrafluorophenol ester monomer with very low polarity into an isocyanoglucosamide copper salt precursor of this kind with relatively low polarity and easy to separate and purify.

[0073] Example 2

[0074] Select the precursor compound of the glucosamide derivative labeled with general formula II in this application - the copper isocyanide-labeled precursor 30 (listing the left part of general formula II and general formula In this case, a is 2 and J is R is -CH 3 , A 1 , A 2 are both H, Q 2 is a cuprous ion, E 2 is a tetrafluoroborate ion), and the Re complexes (standards) 31 - 33 of some typical compounds and their corresponding radioactive 99m Tc complexes 34 - 36 labeling routes and methods (listing the case where a is 2, b is 3, 5, 7, J is R is -CH 3 , and z is 1) are as follows:

[0075] (1) The synthesis and labeling routes are as follows:

[0076] Synthesis route of the labeling precursor:

[0077]

[0078] Re complexes (standards) and their corresponding radioactive 99m Tc complex labeling route:

[0079]

[0080] (2) The synthesis and labeling process are as follows:

[0081] Synthesis of intermediate 26: Dissolve levulinic acid (raw material 22, 17.0000 g), DPPA (raw material 23, 36.7500 g) and triethylamine (19.3500 ml) in toluene. After stirring the mixture at room temperature for 30 min, reflux and stir for 15 min to obtain the highly chemically active intermediate 24. Without separation, directly add benzyl alcohol (raw material 25, 22.5000 g) to it, then reflux and stir the mixture at 80 °C overnight. Concentrate the mixture under vacuum, and purify the concentrate by silica gel column chromatography to obtain intermediate 26 (10.1000 g), and its characterization data: 1 H NMR (400 MHz, DMSO - d 6 ) δ 7.29 (q, J = 6.9 Hz, 5H), 7.17 (t, J = 6.0 Hz, 1H), 4.96 (s, 2H), 3.15 (q, J = 6.9, 6.1 Hz, 2H), 2.55 (t, J = 6.8 Hz, 2H), 2.04 (s, 3H).

[0082] Synthesis of Intermediate 27: To a solution of Intermediate 26 (10.0000 g) in ethyl acetate and ethanol (the ratio between ethyl acetate and ethanol is relatively free, such as the volume ratio is usually 1:(0.01 - 30), here it is 1:1), 10% Pd / C (4.0000 g) and 1.2 mol / L hydrochloric acid (42.5000 ml) were added. Hydrogen was introduced and the mixture was stirred at normal temperature and pressure for 24 h. After the mixture was concentrated under vacuum, the concentrate was purified by silica gel column chromatography to obtain Intermediate 27 (10.1000 g), and its characterization data: 1 HNMR(400MHz,CD 3 OD)δ3.13(t,J=6.5Hz,2H),2.90 - 2.96(m,2H),2.18(s,3H).

[0083] Synthesis of Intermediate 28: To a dichloromethane solution of Intermediate 27 (2.0000 g), triethylamine (1.8000 g) was added. Then, freshly prepared ethyl formate anhydride (1.9000 mL, for the specific operation, refer to the synthesis of Intermediate 4 in Example 1) was added portionwise at 0 °C. After the mixture was concentrated under vacuum, water was added. The aqueous phase was directly extracted with ethyl acetate, and the organic phase concentrate was purified by silica gel column chromatography to obtain Intermediate 28, and its characterization data: 1 H NMR(400MHz,CDCl 3 )δ8.06(s,1H),3.48(dt,J=21.9,6.6Hz,2H),2.72(dt,J=20.9,6.6Hz,2H),2.15(d,J=6.6Hz,3H). The above mixture can also be directly concentrated under vacuum and used directly in the next reaction without further purification.

[0084] Synthesis of Intermediate 29: To a dichloromethane solution of Intermediate 28 (crude product mass 0.5000 g), triethylamine (1.8300 mL) was added. Then, phosphorus oxychloride (0.5000 mL) was added portionwise at 0 °C. The mixture was first stirred at 0 °C for 40 min and then at room temperature for 30 min. The mixture was quenched with a 20% aqueous potassium carbonate solution, extracted with dichloromethane. The combined organic phases were dried, filtered by suction and concentrated under vacuum. The obtained concentrate was purified by silica gel column chromatography to obtain Intermediate 29 (0.2000 g), and its characterization data: 1 HNMR(400MHz,CDCl 3 )δ3.63(tt,J=6.8,2.0Hz,2H),2.87(tt,J=6.7,1.9Hz,2H),2.21(s,3H).

[0085] Synthesis of isocyanide cuprous salt labeled precursor 30: For the specific operation, refer to Example 1 "Synthesis of isocyanide tetrafluorophenol ester cuprous salt intermediate 15". Intermediate 29 (0.2000 g) was reacted with copper(I) tetra(acetonitrile) tetrafluoroborate (161.9 mg) to obtain isocyanide cuprous salt labeled precursor 30 (75.1 mg). Its characterization data: 1 H NMR (400 MHz, CD 3 OD) δ 3.81 (t, J = 6.2 Hz, 8H), 2.98 (t, J = 5.7 Hz, 8H), 2.17 (s, 12H); 13 C NMR (101 MHz, CD 3 OD) δ 205.24, 40.94, 36.37, 28.28; HRMS: calcd. for C 10 H 14 CuN 2 O 2 + 257.0346 (2M + Cu) + , found 257.0391 (2M + Cu) + 。

[0086] Labeling and separation of Re complex 31: Based on the small molecular weight isocyanide cuprous salt labeled precursor 30 (1 mg, 5.00 equivalents) in this example, isocyanide glucosamide cuprous salt labeled precursor 19 (2.00 equivalents), ammonium perrhenate (36.00 equivalents), stannous chloride dihydrate (36.00 equivalents) in Example 1 and a freeze-dried blank kit were added. The mixture was dissolved in pure water or physiological saline and reacted at 100 °C for 30 min. After cooling, the mixture was filtered, and the filtrate was separated by HPLC (high performance liquid chromatography) C-18 reversed-phase semi-preparative column to obtain Re complex 31 (methanol / water containing one-thousandth trifluoroacetic acid = 33 / 67, total flow rate 0.5 mL / min, C-18 reversed-phase semi-preparative column, retention time 88.835 min). Its characterization data; HRMS: calcd. for C 36 H 53 N 7 O 11 Re + 946.3355 (M) + , found 946.3363 (M) + 。

[0087] Labeling and separation of Re complex 32: For the specific operation, refer to "Labeling of Re complex 31". Using the small-molecular-weight isocyanocopper salt labeling precursor 30 in this example, the isocyanoglucosamide copper salt labeling precursor 20 in Example 1, ammonium perrhenate, stannous chloride dihydrate, and one of the above freeze-dried blank cartridges, Re complex 32 was obtained (methanol / water containing one-thousandth trifluoroacetic acid = 36 / 64, total flow rate 0.5 mL / min, C-18 reverse-phase semi-preparative column, retention time 86.010 min). Its characterization data: HRMS: calcd. for C 38 H 57 N 7 O 11 Re + 974.3668(M) + ,found 974.3658(M) + 。

[0088] Labeling and separation of Re complex 33: For the specific operation, refer to "Labeling of Re complex 31". Using the small-molecular-weight isocyanocopper salt labeling precursor 30 in this example, the isocyanoglucosamide copper salt labeling precursor 21 in Example 1, ammonium perrhenate, stannous chloride dihydrate, and one of the above freeze-dried blank cartridges, Re complex 32 was obtained (methanol / water containing one-thousandth trifluoroacetic acid = 38 / 62, total flow rate 0.5 mL / min, C-18 reverse-phase semi-preparative column, retention time 85.368 min). Its characterization data: HRMS: calcd. for C 40 H 61 N 7 O 11 Re + 1002.3981(M) + ,found 1002.3984(M) + 。

[0089] 99m Labeling of Tc complex 34: In this example, since the labeled technetium 99m Tc complex is radioactive and its mass spectrum cannot be directly measured in most cases, usually the corresponding Re complex is first synthesized, its mass spectrum and liquid-phase retention time are measured, etc. If the product is okay, then the technetium 99m Tc complex is prepared by the same method, then its liquid-phase retention time is measured, and the corresponding Re complex and the technetium 99m Tc-labeled target product structure. The same method is also used to confirm in other examples of the present invention.

[0090] Based on the small-molecular-weight isocyanocopper salt labeling precursor 30 (1 mg, 5.00 equivalents) in this example, the isocyanoglucosamide copper salt labeling precursor 19 (2.00 equivalents) in Example 1 and a lyophilized blank kit were added. The mixture was dissolved in pure water or physiological saline, and then Na 99m TcO 4 saline eluate (370 - 3700 MBq, this eluate is from the Mo-Tc generator of Beijing Senke Pharmaceutical Co., Ltd.) was added, and the reaction was carried out at 100 °C for 30 min. After cooling, the mixture was filtered, and the filtrate was separated by HPLC (high-performance liquid chromatography) C-18 reversed-phase semi-preparative column to obtain 99m Tc radioactive complex 34 (methanol / water containing one-thousandth trifluoroacetic acid = 33 / 67, total flow rate 0.5 mL / min, C-18 reversed-phase semi-preparative column, elution time was 88.319 min). Then 99m the elution time of Tc radioactive complex 34 (88.319 min) was basically the same as that of the corresponding Re complex 31 (89.835 min).

[0091] 99m Labeling of Tc complex 35: For the specific operation, refer to " 99m Labeling of Tc complex 34". Using the small-molecular-weight isocyanocopper salt labeling precursor 30 in this example, the isocyanoglucosamide copper salt labeling precursor 20 in Example 1 and a lyophilized blank kit as described above, and then adding Na 99m TcO 4 saline eluate (370 - 3700 MBq, this eluate is from the Mo-Tc generator of Beijing Senke Pharmaceutical Co., Ltd.), 99m Tc radioactive complex 35 (methanol / water containing one-thousandth trifluoroacetic acid = 36 / 64, total flow rate 0.5 mL / min, C-18 reversed-phase semi-preparative column, elution time was 85.466 min) was obtained. Then 99m the elution time of Tc radioactive complex 35 (85.466 min) was basically the same as that of the corresponding Re complex 32 (86.010 min).

[0092] 99m Labeling of Tc complex 36: For the specific operation, refer to " 99m Labeling of Tc complex 34". Using the small-molecular-weight isocyanocopper salt labeling precursor 30 in this example, the isocyanoglucosamide copper salt labeling precursor 20 in Example 1 and a lyophilized blank kit as described above, and then adding Na 99m TcO 4Normal saline eluent (370 - 3700 MBq, the eluent is from the Mo - Tc generator of Beijing Senke Pharmaceutical Co., Ltd.) was used to obtain 99m Tc radioactive complex 36 (methanol / water containing one - thousandth trifluoroacetic acid = 38 / 62, total flow rate 0.5 mL / min, C - 18 reversed - phase semi - preparative column, elution time is 84.810 min), then 99m The elution time of Tc radioactive complex 35 (84.810 min) is basically the same as that of the corresponding Re complex 32 (85.368 min).

[0093] For this example, when synthesizing intermediate 28, some people adopted the following synthetic route:

[0094]

[0095] In this synthetic route, acetic anhydride and metallic sodium are both strictly controlled hazardous chemicals. There are not only great potential safety hazards, but also they are very difficult to buy now. And for the raw materials in the second - step reaction, butenone and formamide both require complex and strict dehydration and distillation purification, which is very inconvenient. Even if the dehydration is complete, when metallic sodium is added, it is extremely easy to release heat. Even when metallic sodium is added extremely slowly under the condition of introducing nitrogen, due to the high chemical reactivity of butenone, the product system of the reaction is extremely complex, which is very unfavorable for the separation of intermediate 28 and the accumulation of its quantity. And if no separation is carried out, the next - step reaction cannot proceed. For these deficiencies, in the synthesis of this application, the inventor creatively adopted a new route. This route does not use such difficult - to - buy hazardous chemicals at all. The effective ingredient of the generated intermediate 28 is high, and for the reaction step of generating intermediate 28, without separation, it still does not affect the generation of isocyano monomer intermediate 29 in the next - step reaction. In this way, the feasibility of the synthesis of such labeling precursors can be greatly enhanced, and the intermediate links of the reaction can be significantly saved.

[0096] Example 3

[0097] The corresponding radioactivity of some typical compounds of general formula II in this application was selected 99m Tc complexes 34 - 36 were studied for their in - vivo biodistribution in female S180 - tumor - bearing mice, as shown below:

[0098] (1) The experimental method is as follows:

[0099] Before the experiment, female S180 - tumor - bearing mice (20 - 22 g, n = 5) with tumors growing for about 1 week were fasted for 12 h. The purified radioactive 99mThe Tc complexes 34 - 36 were respectively formulated into physiological saline solutions of approximately 70 μCi / mL. 100 μL of the above solutions were respectively taken and diluted to 10 mL as %ID. By injecting 100 μL of the above solutions into the caudal veins of female Kunming mice, the mice were decapitated at different time points after caudal vein injection, and blood, brain, heart, liver, spleen, lung, kidney, muscle, bone, large intestine, small intestine, stomach, tumor and tail were collected, weighed and counted, and the counting distribution of each tissue and organ was calculated (unit: %ID / g, the units of the large intestine, small intestine and stomach are %ID), and the data are the average ± standard deviation of five mice in each time phase.

[0100] (2) Radioactivity 99m The in - vivo biodistribution results of the Tc complexes 34 - 36 in female Kunming mice are as follows:

[0101] Table 1 Radioactivity 99m The biodistribution data of the Tc complex 35 in female S180 - tumor - bearing mice (n = 5, ID% / g)

[0102]

[0103] Radioactivity 99m The absolute uptake values of the Tc complex 35 in the tumor tissues of female S180 - tumor - bearing mice at 30, 60 and 120 min after caudal vein injection were respectively (1.81 ± 0.26), (1.61 ± 0.10) and (1.56 ± 0.29) %ID / g. As time passed after caudal vein injection, its value decreased very slowly. Compared with the previously best - performing isocyanoglucose amide - based radioactive 99m Tc complex - 99m Tc(CN5DG) 6 + (Patent Application No.: 201710451094.8), compared with 99m Tc(CN5DG) 6 + The absolute uptake values of 99m Tc(CN5DG) in the tumor tissues of female S180 - tumor - bearing mice at 30, 60 and 120 min after caudal vein injection were respectively (1.83 ± 0.18), (1.07 ± 0.21) and (0.75 ± 0.07) %ID / g], at 60 and 120 min after caudal vein injection, the tumor uptake values of the radioactive 99m Tc complex 35 of the present invention were significantly higher than those of - 6 + Tc(CN5DG). During the period from 30 to 120 min after caudal vein injection, the tumor retention of the radioactive 99m Tc complex 35 of the present invention was significantly better than that of - 99m Tc(CN5DG)6 + is somewhat prolonged. Moreover, the radioactivity of the present invention 99m The myocardial uptake values of Technetium complex 35 at 30, 60, and 120 min after tail vein injection are also significantly higher than those of 99m Tc(CN5DG) 6 + the myocardial uptake values [at 30, 60, and 120 min after tail vein injection, 99m Tc(CN5DG) 6 + the absolute myocardial uptake values in female S180 tumor-bearing mice are (0.64 ± 0.12), (0.41 ± 0.02), and (0.22 ± 0.04) %ID / g respectively], indicating that the drug of the present invention may very likely have a certain degree of glucose metabolism property. This shows that reducing the molecular weight of the complex may have played a role.

[0104] Table 2 Biodistribution data of radioactive 99m Technetium complex 36 in female S180 tumor-bearing mice (n = 5, ID% / g)

[0105]

[0106]

[0107] Radioactivity 99m After tail vein injection of Technetium complex 36, with the passage of time, the uptake value of the tumor tissue in female S180 tumor-bearing mice shows a gradually decreasing trend, but compared with 99m Technetium complex 35, indicators such as the absolute tumor uptake value and tumor / blood value are significantly lower than 99m Technetium complex 35.

[0108] Table 3 Biodistribution data of radioactive 99m Technetium complex 34 in female S180 tumor-bearing mice (n = 5, ID% / g)

[0109]

[0110] Radioactivity 99m Technetium complex 34 compared with 99m Technetium complex 35, at 120 min after tail vein injection, the absolute tumor uptake value and tumor / blood value are significantly lower than 99m Technetium complex 35, but in terms of tumor / muscle and tumor / bone values, it is superior to 99m Technetium complex 35.

[0111] Example 4

[0112] Select the labeled precursors for the left part of the general formula III glucosamide compounds - isocyanocopper salts labeled precursors 54 - 57 and 61 (labeled precursors 54 - 57 correspond to the left part of the general formula II and the general formula where a is 2 - 5 and J is R, R 1 are both -H, A 1 and A 2 are both H, Q 2 is cuprous ion, E 2 is the case of tetrafluoroborate ion; labeled precursor 61 corresponds to the left part of the general formula II and the general formula where a is 2 and J is R is -CH 3 , R 1 is -H, A 1 and A 2 are both H, Q 2 is cuprous ion, E 2 is the case of tetrafluoroborate ion).

[0113] (1) The synthesis and labeling routes are as follows

[0114]

[0115] (2) The synthesis process is as follows:

[0116] Synthesis of intermediate 42: React raw material 37 (5.0000 g) and ethyl formate (raw material 41, 123.2612 g) under reflux with stirring overnight. After concentrating the mixture under vacuum to remove the excess raw material 41, intermediate 42 (8.5359 g) is obtained and directly used in the next reaction without further purification. Its characterization data: 1 H NMR (400 MHz, DMSO - d 6 ) δ 7.83 - 8.18 (m, 4H), 3.13 - 3.16 (m, 4H).

[0117] Synthesis of intermediate 43: Refer to the "Synthesis of intermediate 42". React raw material 38 (5.0000 g) and raw material 41 (99.9326 g) to obtain intermediate 42 (7.2036 g), which is directly used in the next reaction without further purification. Its characterization data: 1 H NMR (400 MHz, DMSO - d 6 ) δ 7.92 - 8.02 (m, 4H), 3.11 (q, J = 7.0 Hz, 4H), 1.56 (p, J = 7.0 Hz, 2H).

[0118] Synthesis of Intermediate 44: Referring to "Synthesis of Intermediate 42", raw material 39 (5.0000 g) and raw material 41 (84.0386 g) were reacted to obtain Intermediate 44 (7.0258 g), which was directly used in the next reaction without further purification.

[0119] Synthesis of Intermediate 45: Referring to "Synthesis of Intermediate 42", raw material 40 (5.0000 g) and raw material 41 (72.4995 g) were reacted to obtain Intermediate 44 (6.1252 g), which was directly used in the next reaction without further purification.

[0120] Synthesis of Intermediate 46: At 0 °C, phosphorus oxychloride (11.0910 g) was added portionwise to a dichloromethane solution of Intermediate 42 (3.0000 g) and triethylamine (26.1428 g), and then the mixture was stirred at 0 °C for 2 - 5 h. The mixture was poured into ice water (136.4887 mL) containing anhydrous potassium carbonate (27.2977 g) for quenching, and then extracted with dichloromethane. The organic phase was dried over anhydrous sodium sulfate, filtered by suction, and the filtrate was concentrated under vacuum. The concentrate was separated by silica gel column chromatography to obtain Intermediate 46 (1.4362 g), and its characterization data: 1 H NMR (400 MHz, CDCl 3 ) δ 3.72 (s, 4H).

[0121] Synthesis of Intermediate 47: Referring to "Synthesis of Intermediate 46", Intermediate 43 (3.0344 g), triethylamine (23.5921 g) and phosphorus oxychloride (10.0089 g) were reacted to obtain Intermediate 47 (1.5378 g), and its characterization data: 1 H NMR (400 MHz, CDCl 3 ) δ 3.63 (t, J = 6.8 Hz, 4H), 2.04 (s, 2H).

[0122] Synthesis of Intermediate 48: Referring to "Synthesis of Intermediate 46", Intermediate 44 (5.0000 g), triethylamine (35.0940 g) and phosphorus oxychloride (14.8885 g) were reacted to obtain Intermediate 48 (2.6225 g), and its characterization data: 1 H NMR (400 MHz, CDCl 3 ) δ 3.49 (t, J = 2.8 Hz, 4H), 1.87 (s, 4H).

[0123] Synthesis of Intermediate 49: Referring to "Synthesis of Intermediate 46", Intermediate 45 (5.0000 g), triethylamine (31.9817 g) and phosphorus oxychloride (13.5681 g) were reacted to obtain Intermediate 49 (2.5400 g), and its characterization data: 1¹H NMR (400 MHz, CDCl 3 ) δ 3.39 - 3.49 (m, 4H), 1.69 - 1.80 (m, 4H), 1.59 - 1.66 (m, 2H).

[0124] Synthesis of Intermediate 50: After adding 0.5 mol / L aqueous citric acid solution (47.8000 mL) to the aqueous solution containing Intermediate 46 (727.0 mg), the mixture was stirred at room temperature for 40 min, quenched with an excess of saturated sodium bicarbonate solution, extracted with ethyl acetate. After the organic phase was dried and filtered by suction, the filtrate was concentrated under vacuum. The concentrate was separated by silica gel column chromatography to obtain Intermediate 50 (108.5 mg), and its characterization data: 1 ¹H NMR (400 MHz, CDCl 3 ) δ 8.24 (s, 1H), 6.70 (brs, 1H), 3.54 - 3.62 (m, 4H).

[0125] Synthesis of Intermediate 51: Referring to the "Synthesis of Intermediate 50", 0.5 mol / L aqueous citric acid solution (38.3770 mL) and Intermediate 47 (686.3 mg) reacted to obtain relatively pure Intermediate 51 (160.0 mg), and its characterization data: 1 ¹H NMR (400 MHz, CDCl 3 ) δ 8.11 (s, 1H), 6.66 (brs, 1H), 3.34 - 3.45 (m, 4H), 1.83 - 1.91 (m, 2H); 13 ¹³C NMR (101 MHz, CDCl 3 ) δ 161.94, 39.27, 39.20, 39.14, 34.63, 29.64, 28.84.

[0126] Synthesis of Intermediate 52: Referring to the "Synthesis of Intermediate 50", 0.5 mol / L aqueous citric acid solution (48.6800 mL) and Intermediate 48 (1.0000 g) reacted to obtain relatively pure Intermediate 52 (250.3 mg), and its characterization data: 1 ¹H NMR (400 MHz, CDCl 3 ) δ 8.19 (s, 1H), 5.88 (brs, 1H), 3.40 - 3.50 (m, 2H), 3.27 - 3.40 (m, 2H), 1.66 - 1.79 (m, 4H).

[0127] Synthesis of Intermediate 53: Referring to the "Synthesis of Intermediate 50", 0.5 mol / L aqueous citric acid solution (47.0600 mL) and Intermediate 49 (1.0917 g) were reacted to obtain relatively pure Intermediate 53 (57.1 mg), and its characterization data: 1 H NMR (400 MHz, CDCl 3 ) δ 8.17 (s, 1H), 6.13 (brs, 1H), 3.37 - 3.48 (m, 2H), 3.17 - 3.36 (m, 2H), 1.65 - 1.82 (m, 2H), 1.44 - 1.65 (m, 4H).

[0128] Synthesis of Isocyanide Cuprous Salt Labeling Precursor 54: For the specific operation, refer to Example 1 "Synthesis of Isocyanotetrafluorophenol Ester Cuprous Salt Intermediate 15". Intermediate 50 (108.5 mg) was reacted with Copper(I) Tetrakis(acetonitrile) Tetrafluoroborate (87.0 mg), and Isocyanide Cuprous Salt Labeling Precursor 54 (43.2 mg) precipitated from the reaction mixture. Its characterization data: 1 H NMR (400 MHz, CD 3 OD) δ 8.19 (s, 4H), 3.82 (t, J = 5.8 Hz, 8H), 3.57 (t, J = 5.8 Hz, 8H); 13 C NMR (101 MHz, CD 3 OD) δ 163.08, 42.28, 36.22; 19 F NMR (376 MHz, CD 3 OD) δ -154.51.

[0129] Synthesis of Isocyanide Cuprous Salt Labeling Precursor 55: For the specific operation, refer to Example 1 "Synthesis of Isocyanotetrafluorophenol Ester Cuprous Salt Intermediate 15". Intermediate 51 (160.0 mg) was reacted with Copper(I) Tetrakis(acetonitrile) Tetrafluoroborate (112.2 mg), and Isocyanide Cuprous Salt Labeling Precursor 55 (78.6 mg) precipitated from the reaction mixture. Its characterization data: 1 H NMR (400 MHz, CD 3 OD) δ 8.11 (s, 4H), 3.73 (t, J = 6.5 Hz, 8H), 3.37 (t, J = 6.7 Hz, 8H), 1.95 (p, J = 6.8, 6.3 Hz, 8H); 13 C NMR (101 MHz, CD 3 OD) δ 163.05, 40.19, 34.25, 28.13; 19 F NMR (376 MHz, CD 3 OD) δ -154.49.

[0130] Synthesis of isocyanide copper salt labeling precursor 56: For the specific operation, refer to Example 1 "Synthesis of isocyanotetrafluorophenol ester copper salt intermediate 15". Intermediate 52 (250.3 mg) was reacted with copper(I) tetra(acetonitrile) tetrafluoroborate (156.0 mg) to obtain relatively pure isocyanide copper salt labeling precursor 56 (107.2 mg). Its characterization data: 1 H NMR(400MHz,CD 3 OD)δ8.08(s,1H),3.72(t,J=6.6Hz,2H),3.29(t,J=6.0Hz,2H),1.79(p,J=6.1,4.8Hz,2H),1.57-2.70(m,2H); 13 CNMR(101MHz,CD 3 OD)δ162.59,36.03,25.97,24.30; 19 F NMR(376MHz,CD 3 OD)δ-155.50.

[0131] Synthesis of isocyanide copper salt labeling precursor 57: For the specific operation, refer to Example 1 "Synthesis of isocyanotetrafluorophenol ester copper salt intermediate 15". Intermediate 52 (57.1 mg) was reacted with copper(I) tetra(acetonitrile) tetrafluoroborate (32.0 mg) to obtain relatively pure isocyanide copper salt labeling precursor 57 (67.4 mg). Its characterization data: 1 H NMR(400MHz,CD 3 OD)δ8.05(s,3H),3.69(t,J=6.6Hz,8H),3.25(t,J=6.9Hz,8H),1.78(p,J=7.5Hz,8H),1.53-1.63(m,8H),1.43-1.53(m,8H); 13 C NMR(101MHz,CD 3 OD)δ162.46,36.38,28.03,27.87,23.25; 19 F NMR(376MHz,CD 3 OD)δ-155.44.

[0132] Synthesis of intermediate 59: Referring to "Synthesis of intermediate 42", raw material 58 (1.0516 g) and raw material 41 (15.2541 g) were reacted to obtain intermediate 59 (1.3102 g), which was directly used in the next step without further purification. Its characterization data: 1 H NMR(400MHz,CDCl 3) δ 8.16 (s, 1H), 7.31 (s, 1H), 7.11 (s, 1H), 3.33 - 3.45 (m, 4H), 1.99 (s, 3H).

[0133] Synthesis of Intermediate 60: Referring to "Synthesis of Intermediate 46", raw material 43 (1.3102 g), triethylamine (2.8217 g) and phosphorus oxychloride (1.6978 g) were reacted to obtain Intermediate 60 (756.4 mg), and its characterization data: 1 H NMR (400 MHz, CDCl 3 ) δ 6.61 (s, 1H), 3.57 (t, J = 5.6 Hz, 2H), 3.44 - 3.52 (m, 2H), 2.03 (s, 3H); 13 C NMR (101 MHz, CDCl 3 ) δ 170.95, 53.49, 41.74, 41.67, 41.60, 38.74, 23.00.

[0134] Synthesis of Isocyanocopper Salt Labeling Precursor 61: For the specific operation, refer to Example 1 "Synthesis of Isocyanotetrafluorophenol Ester Copper Salt Intermediate 15". Intermediate 60 (756.4 mg) was reacted with copper(I) tetra(acetonitrile) tetrafluoroborate (530.5 mg) to obtain copper salt labeling precursor 61 (372.3 mg), and its characterization data: 1 H NMR (400 MHz, CD 3 OD) δ 3.79 (t, J = 5.7 Hz, 8H), 3.51 (t, J = 5.9 Hz, 8H), 2.01 (s, 12H); 13 C NMR (101 MHz, CD 3 OD) δ 172.44, 46.99, 42.41, 42.34, 42.28, 37.68, 21.22; 19 F NMR (376 MHz, CD 3 OD) δ -155.16.

[0135] Example 5

[0136] Select the labeling precursors of the left part of the general formula IV glucosamide compounds in this application - isocyanocopper salt labeling precursors 67 - 69 (labeling precursor 67 corresponds to listing the left part of the general formula IV and the general formula where a is 2, J is R, R 2 are both -H, A 1 and A 2 are both H, Q 2 is a copper ion, E 2For the case of tetrafluoroborate ion; the labeled precursors 68 - 69 correspond to the left part of the general formula IV and the general formula where a is 1 - 2 and J is R is -CH 3 , R 2 is -H, A 1 and A 2 are both H, Q 2 is cuprous ion, E 2 is for the case of tetrafluoroborate ion), and the Re complexes (standards) 70 of some typical compounds and their corresponding radioactive 99m Tc complexes 71 labeling routes and methods (listing the case where a is 2, b is 7, and J is R, R 2 are both -H, A 1 and A 2 are both H, and z is 1).

[0137] (1) The synthesis and labeling routes are as follows:

[0138] Synthesis route of the labeled precursor:

[0139]

[0140] Re complexes (standards) and their corresponding radioactive 99m Tc complex labeling route:

[0141]

[0142] (2) The synthesis process is as follows:

[0143] Synthesis of intermediate 64: The raw material 63 (5.6600 g) and a 2 mol / L methanol solution of ammonia (50.0353 mL) were stirred overnight at room temperature. After the mixture was concentrated under vacuum, the concentrate was separated by silica gel column chromatography to obtain pure intermediate 64 (1.5600 g), and its characterization data: 1 H NMR (400 MHz, DMSO-d 6 ) δ 7.46 (brs, 1H), 7.00 (brs, 1H), 3.61 - 3.67 (m, 2H), 2.40 - 2.46 (m, 2H).

[0144] Synthesis of intermediate 65: The raw material 62 (1.0000 g) and a 2 mol / L methanol solution of methylamine (45.0000 mL) were stirred overnight at room temperature. After the mixture was concentrated under vacuum, the concentrate was separated by silica gel column chromatography to obtain pure intermediate 65 (700.0 mg), and its characterization data: 1HNMR (400 MHz, CDCl 3 ) δ 6.60 (s, 1H), 4.14 (s, 2H), 2.87 (d, J = 4.8 Hz, 3H); 13 C NMR (101 MHz, CDCl 3 ) δ 163.36, 162.41, 45.38, 45.31, 45.24, 26.66.

[0145] Synthesis of Intermediate 66: Raw material 63 (1.0000 g) and a methanol solution of 2 mol / L methylamine (45.0000 mL) were stirred overnight at room temperature. After the mixture was concentrated under vacuum, the concentrate was separated by silica gel column chromatography to obtain pure Intermediate 66 (800.0 mg). Its characterization data: 1 HNMR (400 MHz, CDCl 3 ) δ 5.96 (s, 1H), 3.71 (tt, J = 6.8, 1.9 Hz, 2H), 2.81 (d, J = 4.8 Hz, 3H), 2.54 (tt, J = 6.8, 2.2 Hz, 2H); 13 C NMR (101 MHz, CDCl 3 ) δ 169.12, 156.82, 156.76, 156.71, 38.01, 37.94, 37.88, 35.90, 26.48.

[0146] Synthesis of Isocyanide Copper Salt Precursor 67: For the specific operation, refer to Example 1 "Synthesis of Isocyanide Tetrafluorophenol Ester Copper Salt Intermediate 15". Intermediate 64 (1.0000 g) was reacted with copper(I) tetra(acetonitrile) tetrafluoroborate (801.5 mg), and the copper salt precursor 67 (700.0 mg) was directly precipitated from the reaction mixture. Its characterization data: 1 HNMR (600 MHz, D 2 O) δ 8.13 (s, 4H), 4.60 (s, 8H), 2.60 (s, 12H); HRMS (High Resolution): calcd. for C 8 H 12 CuN 4 O 2 + 259.0251 (2M + Cu) + , found 259.0241 (2M + Cu) + .

[0147] Synthesis of isocyanocopper salt labeling precursor 68: For the specific operation, refer to Example 1 "Synthesis of isocyanotetrafluorophenol ester copper salt intermediate 15". Intermediate 65 (700.0 mg) was reacted with copper(I) tetra(acetonitrile) tetrafluoroborate (561.1 mg), and isocyanocopper salt labeling precursor 68 (413.0 mg) was directly precipitated from the reaction mixture. Its characterization data: 1 HNMR(600MHz,DMSO-d 6 )δ3.75(t,J=6.1Hz,8H),2.63(t,J=4.0Hz,8H); 13 C NMR(101MHz,DMSO-D 6 )δ170.80,39.64,39.43,34.17.

[0148] Synthesis of isocyanocopper salt labeling precursor 69: For the specific operation, refer to Example 1 "Synthesis of isocyanotetrafluorophenol ester copper salt intermediate 15". Intermediate 66 (800.0 mg) was reacted with copper(I) tetra(acetonitrile) tetrafluoroborate (561.1 mg), and isocyanocopper salt labeling precursor 69 (485.0 mg) was directly precipitated from the reaction mixture. Its characterization data: 1 H NMR(600MHz,CD 3 OD)δ3.87(d,J=7.4Hz,8H),2.74(s,12H),2.62(d,J=7.3Hz,8H); 13 C NMR(101MHz,CD 3 OD)δ170.22,39.10,39.04,38.97,34.08,25.10。

[0149] Labeling and separation of Re complex 70: For the specific operation, refer to "Labeling of Re complex 31" in Example 2. Using the small-molecular-weight isocyanocopper salt labeling precursor 67 in this example, the isocyanoglucosamide copper salt labeling precursor 21 in Example 1, ammonium perrhenate, stannous chloride dihydrate, and a lyophilized blank kit described in Example 2, Re complex 70 was obtained (methanol / water containing one-thousandth trifluoroacetic acid = 25 / 75, total flow rate 0.5 mL / min, C-18 reversed-phase semi-preparative column, retention time 92.948 min). Its characterization data: HRMS: calcd. for C 35 H 56 N 12 O 11 Re + 1007.3744(M) + ,found1007.3719(M) + 。

[0150] 99m Labeling of Tc complex 71: For specific operations, refer to " 99m Labeling of Tc complex 34" in Example 2. It is labeled with a small-molecular-weight isocyanide cuprous salt precursor 67 in this example, an isocyanoglucosamide cuprous salt precursor 21 in Example 1, and a freeze-dried blank kit described in Example 2. Then add and then add Na 99m TcO 4 saline eluate (370 - 3700 MBq, and this eluate comes from the Mo-Tc generator of Beijing Senke Pharmaceutical Co., Ltd.) to obtain 99m Tc radioactive complex 34 (methanol / water containing one-thousandth trifluoroacetic acid = 25 / 75, total flow rate 0.5 mL / min, C-18 reverse-phase semi-preparative column, elution time is 92.241 min). Then 99m the elution time of Tc radioactive complex 71 (92.241 min) is basically the same as that of the corresponding Re complex 70 (92.948 min).

[0151] From the experimental results of Examples 2, 4, and 5, it can be seen that the present invention has synthesized for the first time other small-molecular-weight isocyanide cuprous salt precursors for the left-middle part of isocyanoglucosamide tumor imaging agents. The synthesis steps of such labeling precursors are simple, and most of the labeling precursors can be directly precipitated from the reaction mixture, which greatly reduces the intermediate links of separation and purification, saves the synthesis time of the labeling precursor, reduces the workload, and is conducive to the realization of kitization in future clinical research.

[0152] Example 6

[0153] Select the corresponding radioactive 99m Tc complex 71 of the typical compound in Part Ⅳ of the general formula in this application for in vivo biodistribution study in female S180 tumor-bearing mice. For the specific experimental method, refer to the experimental method of "In vivo biodistribution study of radioactive 99m Tc complexes 34 - 36 in female S180 tumor-bearing mice" in Example 3. The results are as follows:

[0154] Table 4 Biodistribution data of radioactive 99m Tc complex 71 in female S180 tumor-bearing mice (n = 5, ID% / g)

[0155]

[0156] Radioactive 99m Tc complex 71 compared with 99m Tc complex 35, at 60 min and 120 min after tail vein injection, the absolute tumor uptake value, tumor retention, and tumor / blood value are significantly lower than 99mTc complex 35, but is superior in terms of tumor / lung, tumor / muscle, and tumor / bone values 99m to Tc complex 35. Further investigation is required. The poor tumor retention may be due to the presence of a primary amide bond in this compound, which has stronger water solubility than 99m the simple carbonyl group in Tc complex 35 and is more easily metabolized.

[0157] In addition, the synthesis / marking routes and methods of other unlisted products (such as a, b, z are other values not listed in the examples of this application, Q and E are other cations and anions, etc.) are the same as the corresponding steps in the above examples. Due to space limitations, they will not be elaborated here.

[0158] The freeze-dried drug kit or freeze-dried blank drug kit described in this application includes the following components: stannous chloride dihydrate, anhydrous sodium citrate, cysteine hydrochloride monohydrate, and mannitol, and can be directly mixed for preparation. Of course, this drug kit can also be purchased or prepared by other methods according to the prior art. Since this is not the inventive point of this application, this application does not make any restrictions on this.

[0159] The above are only the preferred embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention shall be included within the protection scope of the present invention.

Claims

1. A labeled glucosamine derivative, Characterized in that, The general formula of the derivative is shown in Formula I: In Formula I, M is 99m Tc; z is an integer from 1 to 5, a is an integer from 1 to 5, and b is an integer from 2 to 10; J is or at least one of; X is O; R, R 2 and A 1 -A 4 are independently -H, -CH 3 , -CH 2 CH 3 , -CH 2 CH 2 CH 3 , -CH(CH 3 )CH 3 , -CH 2 CH 2 CH 2 CH 3 , -CH(CH 3 )CH 2 CH 3 , -CH 2 CH(CH 3 )CH 3 , -C(CH 3 ) 3 , -CH 2 CH 2 CH 2 CH 2 CH 3 , -CH(CH 3 )CH 2 CH 2 CH 3 , -CH 2 CH(CH 3 )CH 2 CH 3 , -CH 2 CH 2 CH(CH 3 )CH 3 , -C(CH 3 ) 2 CH 2 CH 3 , -CH 2 C(CH 3 ) 2 CH 3 or -CH(CH 3 )CH(CH 3 )CH 3 .

2. A precursor compound of the labeled glucosamine derivative according to claim 1, Characterized in that, The precursor compound is and / or ; Among them, Q 1 and Q 2 are independently metal cations, and the metal cation is a copper ion or a cuprous ion; E 1 and E 2 are independently anions, and the anion is a tetrafluoroborate ion; A 1 ~A 4 、X, J, a and b are the same as those defined in claim 1; n and m vary according to different cations and anions, as long as they conform to the coordination law, and n and m are the values of 2, 4, and 6.

3. A preparation method of the labeled glucosamine derivative according to claim 1, Characterized in that, The method is as follows: First, synthesize the precursors and ; Then, synthesize the derivative shown in General Formula I by a one-step labeling method; Among them, Q 1 and Q 2 are independently metal cations, and the metal cation is a copper ion or a cuprous ion; E 1 and E 2 are independently anions, and the anion is a tetrafluoroborate ion; A 1 ~A 4 、X, J, a and b are the same as those defined in claim 1; n and m vary according to different cations and anions, as long as they conform to the coordination law, and n and m are the values of 2, 4, and 6.

4. Use of the labeled glucosamine derivative according to any one of claim 1 in the preparation of a tumor imaging agent.

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