TW201345553A - Hepatic cell receptor marker precursor containing trisaccharide and disulfur dinitride ligand, preparation method thereof, contrast agent thereof, and pharmaceutical composition thereof - Google Patents

Abstract

This invention relates to a hepatic cell receptor marker precursor useful for nuclear medicine contrast agents or liver cancer medicine for hepatic cell receptor, which structurally is a bifunctional compound which contains a trisaccharide structure as the first functional group having high affinity to sialoglycoprotein receptor on hepatic cell surface and a disulfur dinitride ligand as the second functional group for forming electro-neutral complex with radioactive isotopes to allow the compound to stay on hepatic cell surface as a marker of radioactive isotopes or to cure liver cancer.

Description

Hepatocyte receptor label precursor containing trisaccharide and diazodisulfide ligand, preparation method thereof, contrast agent thereof and pharmaceutical composition thereof

The present invention relates to a hepatocyte receptor label precursor, a preparation method thereof, a contrast agent thereof, and a pharmaceutical composition thereof, and particularly has a trifunctional structure of a trisaccharide and a diazodisulfide ligand.

Under the influence of the global gradual transition to an aging society and increasing social competition, diseases such as malignant tumors, cerebrovascular diseases, neurological diseases and heart diseases have seriously threatened the health of the world. Therefore, in order to enhance the prevention and treatment of diseases such as early diagnosis and early treatment, and to promote the development of preventive medicine, the functional and molecular diagnosis and treatment techniques of the aforementioned diseases will become a top priority for the development of medical science and technology.
In the prior art, the isotope tracking technology and the serum biochemical marker have been used to detect human diseases or dysfunction, which are safe, non-invasive, convenient and accurate. In the treatment of the subject, the most common field of application is the tracking and treatment of liver disease. In liver diseases, liver fibrosis refers to the pathological process in which hepatic cells have abnormal proliferation of fibrous connective tissue in the presence of necrosis and inflammatory irritation. From the perspective of onset, liver fibrosis is a reversible lesion in the early stage of cirrhosis; cirrhosis is an irreversible result of the further development of liver fibrosis.
Nowadays, the detection and diagnosis methods for liver fibrosis mainly rely on the liver puncture method, but the shortcoming is about four points; first, this method is an invasive detection method; second, this method has causes pain, bleeding, peritonitis and the like. The risk of the disease; third, if the liver tissue is very fragmented or the length of the material is not enough, the doctor will often have errors in the examination and clinical diagnosis of the liver; fourthly, it cannot be repeated clinically to dynamically observe the patient's condition, thus reproducing Low sex.
According to international standards, the degree of liver fibrosis is divided into 4 stages, of which F1 and F2 stages have the best therapeutic effect, which can reverse liver fibrosis and restore the normal liver tissue structure. In F3 stage, the treatment effect will be greatly reduced. Liver fibrosis is not easily reversed; F4 is the most severe stage of liver fibrosis, and it is actually cirrhosis. At this time, the possibility of reversing liver fibrosis is very small, so if it can be correctly found at an early stage The patient can get better treatment results.
Since there are special receptors on human cells that can accept certain proteins or peptides, if this particularity is applied, that is, if specific proteins or peptides are radioactive, these proteins or peptides are used. After entering the human body, it can accumulate in specific organs or tissues, and can achieve the purpose of disease treatment or nuclear diagnostic imaging diagnosis.
In the past, bifunctional ligands have been used to allow ruthenium or osmium compounds to target proteins or peptides. For example, the bifunctional compound S-Hynic contains an activated carboxylic acid that can be used to bind to proteins or peptides. The indoleamine bond, additionally containing a pyridyl group and an azo hydrogen structure, can be used to bond ^99m Tc; when combined with an auxiliary chelating agent such as tris(hydroxymethyl)methylglycine, it can be used with hydrazine or hydrazine. Produces a stable complex. However, the solution of S-Hynic is a light-sensitive substance, which is not very convenient to use, so it is necessary to find a stable bifunctional organic compound.
It is known that there are about 200,000 aspiric acid receptors (ASGPR) on the surface of mammalian hepatocytes. This ASGPR is a liver-specific transmembrane glycoprotein whose main function is to remove sialic glycoprotein and wither Dead cells, as well as clearance of lipoproteins. It has a special affinity for galactose (Gal) and N-acetylgalactosamine (GalNAc), especially when the matrix contains three galactose or N-acetylgalactosamine. The affinity of ASGPR on the surface of hepatocytes is stronger, almost 10 ⁶ times that of a single N-acetylgalactosamine. Based on this characteristic, YEE (ah-GalNAc) ₃ has been applied as a carrier of genes and drugs, and successfully loaded genes and drugs into liver cells.
It is conventionally known that carboxylic acids can bond alcohols, sugars, amines, amino acids, peptides and proteins, and N ₂ S ₂ ligands can bond radioactive isotopes such as ruthenium or osmium, but there is currently no galactose Hepatocyte receptor nuclear medicine contrast agent composed of sugar and N-acetylgalactosamine and N ₂ S ₂ complex.
Because galactose and N-acetylgalactosamine are specific to hepatic lectin, nuclear medicine can bind radioisotopes to galactose and N-acetylgalactosamine. On the glycoprotein, it can successfully locate the liver cells, and can be swallowed into the cells by the liver cells to achieve the purpose of functional angiography or radiation therapy. The design of such glycosylation has not been found in the nuclear medicine industry. Therefore, the present invention proposes a preparation method of a hepatocyte receptor label precursor containing a trisaccharide and a diazodisulfide ligand to enhance the sensitivity and specificity of nuclear medicine detection. Sex.

The main object of the present invention is to provide a hepatocyte receptor label precursor containing a trisaccharide and diazodisulfide complex, a preparation method thereof, a contrast agent thereof and a pharmaceutical composition thereof, which have a dinitrogen The sulfur ligand can chelate the radioactive element and has three activated carboxylic acid esters, which can form a guanamine bond with the amino group-containing compound, thereby becoming a difunctional compound, so that it can bond the polyol on the one hand. Classes, sugars, amines, amino acids, peptides or proteins, which are beneficial to stay on the surface of liver cells; on the other hand, compounds such as TcO ³⁺ or ReO ³⁺ can be bonded, which is very suitable for the development of nuclear medicines. .
A secondary object of the present invention is to provide a hepatocyte receptor label precursor containing a trisaccharide and diazodisulfide complex, a preparation method thereof, a contrast agent thereof, and a pharmaceutical composition thereof, which utilizes trityl group The thiol of the diazo disulfide ligand is protected, and the protecting group can be automatically detached in the case of a mismatch reaction without prior removal, which makes the use more convenient.
Another object of the present invention is to provide a hepatocyte receptor label precursor containing a trisaccharide and a diazodisulfide complex, a preparation method thereof, a contrast agent thereof, and a pharmaceutical composition thereof, which utilize galactoside ah -GalNAc ₄ is a functional group which has a very good affinity with the asialoglycoprotein receptor on hepatocytes, and can ensure the efficacy of liver development and the effect of liver cancer treatment.
Still another object of the present invention is to provide a hepatocyte receptor label precursor containing a trisaccharide and a diazodisulfide complex, a preparation method thereof, a contrast agent thereof, and a pharmaceutical composition thereof, which are synthesized in a semi-milk Glycoside ah-GalNAc ₄ uses benzyloxycarbonyl as the amino-protecting group of 6-aminohexanol instead of trifluoroacetamidine, which ensures that this protecting group is easily detached under hydrogenation and is not hydrogenated. Will affect other functional groups of the molecule.
In order to achieve the above object, the present invention discloses a hepatocyte receptor label precursor containing a trisaccharide and a diazodisulfide complex, a preparation method thereof, a contrast agent thereof and a pharmaceutical composition thereof, and the steps of the preparation thereof are The method comprises: synthesizing one of a difunctional chelating agent having a diazo disulfide ligand; and hydrating the carboxylic acid to form a guanamine which is a difunctional chelating agent having a diazo disulfide ligand. Further, the guanamine has three polycarboxylates by hydrolysis; synthesizing three galactosides; using the galactosides to aminate the polyvalent carboxyl groups of the guanamine to form a trisaccharide-containing diazodisulfide complex Hepatocyte receptor marker precursor. Through these steps, the nuclear medicine contrast agent can be prepared and can be made into a pharmaceutical composition for treating liver cancer.

In order to provide a better understanding and understanding of the features and the efficacies of the present invention, the preferred embodiment and the detailed description are as follows:
In view of the fact that the labeling effect against liver cancer cells in the past is not ideal, or the stability and convenience are not good, the present invention proposes this chemical structure and its preparation method to overcome such problems.
First, please refer to the first figure, which is a chemical structure diagram of a precursor of a hepatocyte receptor marker containing a trisaccharide and a diazodisulfide complex, which has a diazo disulfide ligand structure. In the state where the radioisotope is not chelated, trityl group is used as a protecting group, which is required for protection because the mercaptan is easily oxidized. Before the diazo disulfide ligand structure and the radioisotope compound are mismatched to form an electrically neutral complex, the protecting group of the thiol must be removed first. The trityl group used in the present invention can be automatically detached in the case of a mismatch reaction without being removed in advance, and thus is a convenient choice for use.
In addition to the diazodisulfide ligand structure, the chemical structure of the present invention also has a trisaccharide structure which has high affinity and specificity for ASGPR and can be selectively phagocytized by human hepatoma cell line HepG2. Into the cell, in other words, this structure helps to localize the present invention to liver cancer cells, and is swallowed into cells by liver cancer cells, thereby achieving radioactive labeling or therapeutic purposes.
For the preparation method of the hepatocyte receptor label precursor of the trisaccharide and diazodisulfide ligand disclosed in the present invention, please refer to the second figure; as shown in the figure, the system comprises four basic steps. :
Step S1: synthesizing a carboxylic acid which is a difunctional chelating agent having a diazo disulfide ligand;
Step S2: hydrazine oxidizing the carboxylic acid to form a guanamine which is a difunctional chelating agent having a diazo disulfide ligand, the guanamine further hydrolyzed to have three polycarboxy groups;
Step S3: synthesizing three galactosides;
Step S4: aminating the polyvalent carboxyl groups of the guanamine with the galactoside glycosides to form a hepatocyte receptor label precursor containing a trisaccharide and a diazodisulfide complex.
In these steps, the key technical feature is to establish the structure of the diazo disulfide ligand and the trisaccharide, that is, to synthesize a bifunctional compound having a chelated radioisotope and a high affinity with ASGPR.
In the step S1 of the preparation method of the present invention, the carboxylic acid of the difunctional chelating agent having a diazo disulfide ligand is 7,10-diaza-9-oxo-7-[2- ((tritylmethyl)thio)ethyl]-12-[(trityl)thio]dodecanoic acid, and the synthesis steps of this carboxylic acid are explained in detail in the third figure, the steps are:
Step (1): using 2-thioethylamine hydrochloride and triphenylmethanol, thiol protection reaction under the catalysis of a mixture of boron trifluoride diethyl ether to form a 2-[(trityl)thio] Amine, and the reaction temperature of this thiol protection reaction was 72 ° C, and the reaction time was 4 hours. ;
Step (2): using the 2-[(tritylmethyl)thio]ethylamine and chloroacetic acid chloride to carry out the guanidation reaction in a chloroform solution to form an N-[2-((triphenyl) Methyl)thio)ethyl]chloroacetamide;
Step (3): using the 2-[(trityl)thio]ethylamine and the N-[2-((trityl)thio)ethyl]chloroacetamide, with triethylamine As a reactant, and a substitution reaction in a dichloromethane solvent, a ligand containing an amine-melamine-thiol is formed, and the reaction temperature of the substitution reaction is 55 ° C, and the reaction time is 48 hours;
Step (4): using 6-bromohexanoic acid and sulfinium chloride, esterification reaction in an anhydrous methanol solvent to form a methyl 6-bromohexanoate, and the reaction temperature of the esterification reaction is 25 ° C, the reaction The time is 24 hours;
Step (5): using the amine-melamine-thiol-containing ligand and the 6-bromohexanoic acid methyl ester, using sodium hydroxide as a reactant, performing a substitution reaction in an acetonitrile solution to form a 7,10 -Diazabi-9-oxo-7-[2-((trityl)thio)ethyl]-12-[(trityl)thio]dodecanoate, wherein the substitution The reaction temperature of the reaction is 85 ° C, and the reaction time is 24 hours;
Step (6): hydrolysis of the 7,10-diaza-9-oxo-7-[2-((trityl)thio)ethyl]-12-[(trityl)thio group ] Methyl dodecanoate in an alkaline methanol solution to form the 7,10-diaza-9-oxo-7-[2-((trityl)thio)ethyl]-12-[( Trityl)thio]dodecanoic acid.
After obtaining the carboxylic acid, the carboxylic acid is then aminated in step S2 to form a guanamine type of trimethyl-γ-L-glutamate-L-glutamate-7,10-diaza- 9-oxo-7-[2-((trityl)thio)ethyl]-12-[(trityl)thio]dodecanamine, please refer to the fourth figure for detailed steps. These steps include:
Step (7): using γ-L-glutamic acid-L-glutamic acid and sulfinium chloride to carry out esterification reaction in an anhydrous methanol solvent to form a γ-L-glutamate-L-glutamic acid Trimethyl ester;
Step (8): using the γ-L-glutamate-L-glutamic acid trimethyl ester with the carboxylic acid, and 1,3-dicyclohexylcarbodiimide and N-hydroxybutanediamine The reaction is carried out in a chloroform solution to produce the trimethyl-γ-L-glutamate-L-glutamate-7,10-diaza-9-oxo-7-[ 2-((Triphenylmethyl)thio)ethyl]-12-[(trityl)thio]dodecylamine.
Next, please refer to the fifth figure, this trimethyl-γ-L-glutamate-L-glutamate-7,10-diaza-9-oxo-7-[2-((tritylmethyl) After further hydrolysis of the thio)ethyl]-12-[(trityl)thio]dodecanamine, the compound DODGA having three polyvalent carboxyl groups, which are used for the present invention The moiety bound to galactosidase.
After completion of step S2, the present invention then synthesizes three galactosides in step S3, which is 6'-ethylamino-2-ethinylamino-3,4,6-trioxo-ethenyl-2 -deoxy-β-D-galactoside, a compound ah-GalNAc₄For detailed preparation steps, please refer to the sixth figure, which includes:
Step (9): amino-protective reaction using 6'-aminohexanol with phenyl chlorocarbonate to form a 6'-(N-benzyloxycarbonyl) aminohexanol;
Step (10): reacting with ethyl chlorohydrazine at 10 ° C using N-acetamidine-D-galactose to form 2-ethylamino-2,4,6-trioxo-ethenyl-1-chloro-1 , 2-deoxy-α-D galactopyranosole;
Step (11): using the 6-(N-benzyloxycarbonyl)aminohexanol with the 2-ethylguanidino-2,4,6-trioxo-ethenyl-1-chloro-1,2-deoxy- α-D-galactopyranoside, substituted with methane cyanide as a catalyst in a mixture of toluene and nitromethane to form a 6'-(N-benzyloxycarbonyl)aminohexyl-2-ethyl Amidino-3,4,6-trioxa-ethinyl-2-deoxy-β-D-galactoside;
Step (12): using the 6'-(N-benzyloxycarbonyl)aminohexyl-2-ethenylamino-3,4,6-trioxo-ethenyl-2-deoxy-β-D-galactoside Hydrogenation reduction reaction in ethanol under a palladium carbon catalyst to form the 6'-ethylamino-2-ethinylamino-3,4,6-trioxo-ethenyl-2-deoxy-β-D - galactoside.
In the synthesis of this galactoside, the amino group of galactoside has a wide variety of protecting groups to choose from, and whichever is more appropriate depends on the use of the compound. In order to facilitate the subsequent reaction, the present invention uses a benzyloxycarbonyl group as a protecting group, mainly because the protecting group is easily detached during the hydrogenation reaction, and does not affect other functional groups of the molecule during hydrogenation.
Further, 6'-(N-benzyloxycarbonyl)aminohexyl-2-ethenylamino-3,4,6-trioxa-ethinyl-2-deoxy-β-D produced in the step (11) - Galactose should be isolated and purified using liquid chromatography. The usual method is to separate and purify through a Sephadex LH-20 colloidal filter column. However, the Sephadex LH-20 colloid is quite expensive, so the present invention uses a silicone rubber instead of the Sephadex LH-20 colloid, which is effective and greatly reduces the cost.
And hydrogenation reduction in step (12) to obtain galactoside ah-GalNAc₄After the finished product, in principle, it is no longer necessary to separate and purify. If this galactoside ah-GalNAc₄The purity of the semi-finished product (state before hydrogenation reduction) is high, and only toluene is produced during the hydrogenation. Since toluene is extremely volatile, it should be completely volatilized during the concentration process without residue. But if this galactoside ah-GalNAc₄If the purity of the semi-finished product is insufficient, the product after hydrogenation needs to be separated and purified. At this time, as in the step (11), the liquid chromatography column is prepared by using the tannin extract for separation and purification.
The galactoside ah-GalNAc synthesized here₄It will be combined with the previously prepared compound DODGA having three polyvalent carboxyl groups to form (ah-GalNAc₄)₃- The structure of DODGA, that is, the hepatocyte receptor label precursor of the trisaccharide and diazodisulfide complex of the present invention. Please refer to the seventh figure, the steps of which include:
Step (13): activating the carboxylic acid of the DODGA, followed by the ah-GalNAc₄The amide is reacted with 1,3-dicyclohexylcarbodiimide and N-hydroxybutanediamine as a reagent in a chloroform solution to form a 6-trioxo-(2'-acetamidoamino group- 3',4',6'-Trioxo-acetamido-2'-deoxy-β-D-galactoside)-hexyl-7,10-diaza-9-oxo-7-[2- (trityl)thioethyl]-12-[(trityl)thio]dodecanamine-γ-L-glutamate-L-glutamate;
Step (14): hydrolysis of the 6-trioxo-(2'-acetamido-3',4',6'-trioxo-ethenyl-2'-deoxy-β-D-galactoside)- Hexyl-7,10-diaza-9-oxo-7-[2-((trityl)thio)ethyl]-12-[(trityl)thio]dodecylamine -γ-L-glutamate-L-glutamate under sodium methoxide to form a 6-trioxo-(2'-acetamido-3',4',6'-trihydroxy-2' -deoxy-β-D-galactoside)-hexyl-7,10-diaza-9-oxo-7-[2-(trityl)thioethyl]-12-[(triphenyl) Methyl)thio]dodeamide-γ-L-glutamate-L-glutamate, which is a compound (ah-GalNAc)₄)₃-DODGA.
In the practical application of the present invention, the radioisotope compound MO will be used.³⁺It is bonded to the diazo disulfide ligand as a neutral complex. At this time, the removal of the thiol protecting group on the diazo disulfide ligand is to be (ah-GalNAc)₄)₃-DODGA is dissolved in trifluoroacetic acid, and an excess of triethyl decane is added. Under such operation, the trityl group will be detached from the thiol to form a solid insoluble in trifluoroacetic acid, and the user can filter by It is easy to remove or wash with n-hexane.
Finally, in addition to the hepatocyte receptor marker precursor containing the trisaccharide and diazo disulfide ligand, and the preparation method thereof, the present invention also discloses a pharmaceutical composition which can be applied as a contrast agent or a liver cancer treatment. . Please refer to the eighth figure, which is a chemical structure diagram of a contrast agent or a composition of a pharmaceutical composition of a hepatocyte receptor label precursor containing a trisaccharide and a diazodisulfide complex, wherein M is a radioisotope. Can be^99mTc,¹⁸⁸Re or¹¹¹In etc. can be used for liver cancer signs and treatment elements.
As described above, in addition to the diazodisulfide ligand structure which can chelate a radioisotope, the chemical structure of the present invention has a trisaccharide structure which has high affinity and specificity for ASGPR, and It can be selectively phagocytized into cells by human hepatoma cell line HepG2, so the structure helps to localize the present invention to liver cancer cells, thereby enabling the radioisotope to inhibit or kill the liver cancer cells by means of radiation therapy. The effect of a pharmaceutical composition applied to the treatment of liver cancer. In addition, based on its excellent performance in positioning, it is also a good contrast material for radioactive markers of liver cancer cells, achieving radioactive markers or treating liver cancer.
The compound provided by the invention (ah-GalNAc₄)₃-DODGA contains galactoside ah-GalNAc₄It has a very good affinity with the asialoglycoprotein receptor on hepatocytes, and has a protecting group to protect the thiol before use, has high stability, and it does not have to be removed before forming a complex with the radioisotope. These protective groups are undoubtedly a highly efficient hepatocyte fibrosis nuclear medicine contrast agent or a pharmaceutical composition for treating liver cancer, which has great medical value.
The following are the details of the actual embodiment and related parameters in the experimental operation of the present invention:

[Synthesis of 2-[(trityl)thio]ethylamine]
[2-[(Triphenylmethyl)thio]ethylamine]
2-thioethylamine hydrochloride (5 g, 44.2 mmol), triphenylmethanol (11 g, 42.5 mmol) and triethylamine (7 mL, 49.9 mmol), co-dissolved In chloroform (100 mL). The mixture was heated under reflux, and the mixture was gradually added dropwise to a boron trifluoride ethyl ether complex (15 mL, 119.5 mmol), and the mixture was heated to reflux for 4 hours. After cooling, a sodium hydrogencarbonate aqueous solution was added and stirred, and a white solid precipitated immediately. The solid was taken by suction filtration, washed with water and dried to give 2-[(triphenylmethyl)thio]ethylamine (14.0 g, 99%).
Its analysis data:
IR (neat) n 3381 (NH₂) cm^-1.
¹H NMR (CDCl₃) d 7.42 (m, 3 H, Ph), 7.30 (m, 12 H, Ph), 2.58 (t, J = 6.6 Hz, 2 H, CH₂N), 2.32 (t, J = 6.6 Hz, 2 H, CH₂S), 1.45 (br, 2 H, NH₂).
¹³C NMR (CDCl₃) d 144.80, 192.52, 127.81 and 126.60 (Ph), 66.51 (CPh), 40.94 (CH)₂N), 36.09 (CH₂S).
MS m/z 319 (M⁺), 243 ((CPh₃)⁺)..

[Synthesis of N-[2-((trityl))thio)ethyl] chloroacetamide]
[N-[2-((Triphenylmethyl)thio)ethyl]-
Chloroacetamide
2-[(Trityl)thio]ethylamine (5.24 g, 16.4 mmol) was dissolved in dichloromethane (150 mL) with triethylamine (2.76 mL, 19.6 mmol). Under ice cooling, a solution of chloroacetyl chloride (1.56 mL, 19.6 mmol) in chloroform (20 mL) was slowly added dropwise. After the completion of the dropwise addition, the mixture was stirred at room temperature for 2 hours, and then the organic phase was washed sequentially with 2N aqueous HCl (120 mL) Organic phase via anhydrous sodium sulfate (Na₂SO₄After de-concentration, the residue was evaporated to dryness crystals crystals crystals
Its analysis data:
IR (neat) n 3413 and 3306 (NH), 1662 (CO) cm^-1.
¹H NMR (CDCl₃) d 7.41 (m, 3 H, Ph), 7.24 (m, 12 H, Ph), 6.48 (br, 1 H, NH), 3.97 (s, 2 H, CH₂Cl), 3.12 (q, J = 6.3 Hz, 2 H, CH₂N), 2.43 (t, J = 6.3 Hz, 2 H, CH₂S).
¹³C NMR (CDCl₃) d 165.63 (CO), 144.47, 129.48, 127.97 and 126.81 (Ph), 66.52 (CPh), 42.54 (CH)₂Cl), 38.35 (CH₂N), 31.67 (CH₂S).
MS m/z 397 and 395 (M⁺), 243 ((CPh₃)⁺).

[Synthesis of N-[2-((trityl))thio)ethyl][2-((trityl)thio)ethyl-amino]acetamide]
[N-[2-((Triphenylmethyl)thio)ethyl]]
[2-((triphenylmethyl)thio)ethyl-amino]acetamide]
Take N-[2-((trityl)thio)ethyl]chloroacetamide (5.4 g, 13.8 mmol) with 2-[(trityl)thio]ethylamine (4.4 g, 13.8) Methyl acetate was dissolved in dichloromethane (100 mL), then triethylamine (3.0 mL, 20.8 mmol). After cooling, sodium bicarbonate (NaHCO)₃The aqueous solution (100 mL) was washed and the organic layer was taken. The organic phase was dried over anhydrous sodium sulfate and concentrated and purified using EtOAc EtOAc (EtOAc:EtOAc Ethylthio)ethyl][2-((trityl)thio)ethyl-amino]acetamide (2.2 g, 41.8%).
Its analysis data:
IR (neat) n 3330 (NH), 1670 (CO) cm^-1.
¹H NMR (CDCl₃) d 7.42 (m, 4 H, HNCO and Ph), 7.20 (m, 30 H, Ph), 3.07 (m, 4 H, CH₂NCO and CH₂CO), 2.38 (m, 6 H, CH ₂ NHCH₂CO and CH₂S), 1.94 (br, 1 H, NHCH₂CO).
¹³C NMR (CDCl₃) d 170.84 (CO), 144.61, 129.47, 127.88 and 126.69 (Ph), 66.72 and 66.65 (CPh₃), 51.62 (CH₂CO), 48.19 (CH₂NHCH₂CO), 37.70 (CH₂NHCO), 32.12 and 31.97 (CH₂S).
MS m/z 243 ((CPH₃)⁺)

[Synthesis of methyl 6-bromohexanoate]
[Methyl 6-bromohexadecanoate]
6-bromohexanoic acid (4.1 g, 21.1 mmol) was added to anhydrous methanol (100 mL), and Thionyl chloride (30 mL) was slowly added dropwise in an ice bath, and Stir at room temperature overnight, concentrate and add chloroform to dissolve. The mixture was filtered off with suction, and the filtrate was concentrated to give ethyl 6-bromohexanoate (4.4 g, 100%).
Its analysis data:
IR (neat) n11739 (CO) cm^-1.
¹H NMR (CDCl₃) d 3.64 (s, 3 H, OCH ₃ ), 3.38 (t, 2 H, BrCH ₂ ), 2.30 (t, 2H, CH ₂ COOCH₃), 1.82 (m, 2 H, CH₂CH ₂ CH₂CH₂CH₂), 1.63(m, 2 H, CH₂CH₂CH₂CH ₂ CH₂), 1.46 (m, 2 H, CH₂CH₂CH ₂ CH₂CH₂).
¹³C NMR (CDCl₃) d 173.77 (CO), 51.73 (COOCH₃), 33.72 (BrCH₂), 33.34 (CH₂CH₂CH₂CH₂ CH₂), 32.30 (CH₂ CH₂CH₂CH₂CH₂), 27.56 (CH₂CH₂CH₂ CH₂CH₂), 23.98 (CH₂CH₂ CH₂CH₂CH₂).

[Synthesis of 7,10-diaza-9-oxo-7-[2-((trityl))thio)ethyl]-12-[(trityl)thio]dodecanoate ester】
[Methtyl 7,10-diaza-9-oxo-7-[2-((triphenylmethyl)thio)-
Ethyl]-12-[(triphenylmethyl)thio]dodecanoate]
N-[2-((trityl)thio)ethyl][2-((trityl)thio)ethyl-amino]acetamide (14.2 g, 21.0 mmol) was added to 6- Methyl bromohexanoate (17.6 g, 12.6 mmol) and potassium hydroxide (1.76 g, 31.4 mmol) and acetonitrile (100 mL) were then warmed to reflux overnight. After cooling, it was suction filtered, and the filtrate was concentrated under reduced pressure. The residue was dissolved in methylene chloride (100 mL). The organic phase was separated from water by anhydrous sodium sulfate, and concentrated and purified by liquid chromatography (c.c., ethyl acetate:hexane = 1:1) to give the product 7,10-diaza-9-oxo. Methyl-7-[2-((trityl)thio)ethyl]-12-[(trityl)thio]dodecanoate (7.4 g, 44%).
Its analysis data:
IR (neat) n 2928 (NH), 1735 and 1674 (CO) cm^-1.
¹H NMR (CDCl₃) d 7.40(NH), 7.30 (m, 30H, Ph), 3.64 (s, 3 H, OCH₃), 3.02 (q, 2 H, NHC)H ₂ CH₂S), 2.83 (s, 2 H, CO CH₂N ), 2.36 (m, 4 H, NHCH₂CH ₂ S and NCH ₂ CH₂S ), 2.24 (m, 6 H, NCH₂CH ₂ S and CH ₂ CH₂CH₂CH₂CH ₂ ), 1.53 (m, 2 H, NCH)₂CH ₂ CH₂CH₂CH₂), 1.29 (m, 4 H, CH₂CH₂CH ₂ CH ₂ CH₂).
¹³C NMR (CDCl₃) d 173.90 and 171.13 (CO), 144.68, 144.70, 129.51, 127.86, 126.66, (Ph), 77.43, 77.10 and 76.58 (CPh₃), 66.74 (CH₂N), 58.23 (CH₃O), 54.54 and 53.82 (CH₂S), 51.40 (NHCH₂), 37.89 (NCH₂CH₂S), 33.89 (CH₂CH₂CH₂CH₂CH₂), 31.97 (CH₂CH₂CH₂CH₂ CH₂), 29.94 (CH)₂ CH₂CH₂CH₂CH₂), 26.72 (CH₂CH₂CH₂ CH₂CH₂) , 24.67 (CH₂CH₂ CH₂CH₂CH₂).

[Synthesis of 7,10-diaza-9-oxo-7-[2-((trityl))thio)ethyl]-12-[(trityl)thio]dodecanoic acid]
[7,10-Diaza-9-oxo-7-[2-((triphenylmethyl)thio)-
Ethyl]-12-[(triphnyl-methyl)thio] dodecanoic
Acid]
Potassium hydroxide (20 g) was dissolved in anhydrous methanol (200 mL). Adding methyl 7,10-diaza-9-oxo-7-[2-((trityl)thio)ethyl]-12-[(trityl)thio]dodecanoate (7.4 g, 9.2 mmol) was stirred at room temperature overnight. After concentration under reduced pressure at room temperature, water (30 mL) and methanol (30 mL) were added, and the pH of the reaction mixture was adjusted to pH = 7.0 with concentrated hydrochloric acid, extracted with dichloromethane (2 × 50 mL), and the aqueous phase was discarded. The organic phase layer was taken, the water was removed with anhydrous sodium sulfate and concentrated under reduced pressure to give the product 7,10-diaza-9-oxo-7-[2-((triphenylmethyl)thio)ethyl]-12 -[(Trityl)thio]dodecanoic acid (6.9 g, 94.8%).
Its analysis data:
IR (neat) n 3210 (OH), 2926(NH),1679 (CO) cm^-1.
¹H NMR (CD₃OD) d 7.30 (m, 30H, Ph), 3.07(t, 2 H, COCH ₂ N), 2.84 (m, 2 H, NHCH ₂ CH₂S), 2.75 (m, 2 H, NCH ₂ CH₂S), 2.64 (m, 2 H, NCH₂CH ₂ S), 2.37(t, 2H, NHCH₂CH ₂ S ), 2.28 ( t, 2 H, NCH ₂ CH₂CH₂CH₂CH₂), 1.55 (m, 2 H, CH ₂ COOH ), 1.44 (m, 2 H, CH₂CH₂CH₂CH ₂ CH₂), 1.24(m, 2 H, CH₂CH₂CH ₂ CH₂CH₂).
¹³C NMR (CD₃OD) d 176.94 and 165.29 (CO), 146.03, 145.45, 130.69, 129.28 and 127.81, 128.99, 128.28, 127.90 (Ph), 68.83 and 67.89 (CPh₃), 56 (COCH₂), 55.29 (NCH₂CH₂S), 55.13 (NCH₂CH₂CH₂CH₂CH₂), 39.68 (NHCH₂), 34.44 (NCH₂ CH₂S), 32.55 (NHCH₂ CH₂S), 26.81 (CH₂COOH), 25.29 (CH₂ CH₂CH₂CH₂CH₂), 24.52 (CH₂CH₂ CH₂CH₂CH₂).
MS m/z 243 ((CPh₃)⁺).

[Synthesis of γ-L-glutamate-L-glutamic acid trimethyl ester]
[Methty γ-L-Glutamyl-L-glutamate]
γ-L-Glutamyl-L-glutamic acid (H-Glu(Glu-OH)-OH) (2.0 g, 7.3 mmol) was added to anhydrous methanol (50 mL) In the ice bath, thioxanthene chloride (20 mL) was slowly added dropwise, and stirred at room temperature overnight, and concentrated to give the product γ-L-glutamate-L-glutamic acid trimethyl ester (2.3 g, 100) %).
Its analysis data:
IR (neat) n 3368 (NH₂), 2924 (NH), 1735 and 1657 (CO) cm^-1.
¹H NMR (CDCl₃) d 6.9 (br, NH), 4.52 (m, 1 H, NH₂CH), 3.67 (s, 3 H, OCH₃), 3.65(s, 3 H, OCH₃), 3.44 (s, 3 H, OCH₃) , 3.40 (q, 1 H, NHC)H), 2.32 (q, 2 H, CH₂CH ₂ CONH), 2.00 (m, 4H, CH ₂ CH₂CONH and CH ₂ CH₂COOCH₃), 1.78(m, 2 H, CH₂CH ₂ COOCH₃).
¹³C NMR (CDCl₃) d175.88 , 173.16 and 172.34 (CO), 53.62 (NHCH), 52.35 (NH₂CH), 52.07 (CH₂ CH₂CONH), 51.71 (CH₂ CH₂COO), 51.60(CH₂CH₂CONH), 51.50 (CH₂CH₂COO).
MS m/z 320 ( M⁺-Cl ).

[Synthesis of trimethyl-γ-L-glutamate-L-glutamate-7,10-diaza-9-oxo-7-[2-((trityl)thio)ethyl) ]-12-[(trityl)thio]dodecanamine]
[Trimethyl-γ-L-glutamyl-L-glutamyl-7, 10-Diaza-9-oxo
-7-[2-(triphenylmethyl)thioethyl]-12-
[(triphenylmethyl)-thio]dodecanamide]
Take 7,10-diaza-9-oxo-7-[2-((trityl)thio)ethyl]-12-[(trityl)thio]dodecanoic acid (0.55 g, 0.69 mmol) γ-L-glutamate-L-glutamic acid trimethyl ester (0.22 g, 0.69 mmol) and triethylamine (0.24 mL, 1.7 mmol) and N-hydroxybutanediamine ( N-hydroxysuccinimide) (0.10 g, 0.83 mmol), 1,3-dicyclohexylcarbodiimide (0.2 g, 0.1 mmol) and chloroform (50 mL) at room temperature Stir for 48 hours. The mixture was filtered with suction, and the filtrate was washed with water. The residue was dissolved in acetone (100 mL), filtered, and filtered. The organic phase was separated from water by anhydrous sodium sulfate, concentrated and purified by liquid chromatography (c.c., chloroform:methanol = 95:5) to give the product trimethyl- gamma-L- glutamine-L. - bran 醯-7,10-diaza-9-oxo-7-[2-((trityl)thio)ethyl]-12-[(trityl)thio] decene Diamine (0.31 g, 42.4%).
Its analysis data:
IR (neat) n 3308 and 2927 (NH), 1741 and 1659(CO) cm^-1.
¹H NMR (CDCl₃) d7.46 (br, 1 H, NHCH₂CH₂S), 7.25 (m, 30 H, Ph), d6.80 (br, 1 H, CH₂CH₂CH₂CH₂CH₂CONH), 6.50 (br, 1 H, CH₂CH₂CONH), 4.57(m, 2H, NHCHCOOH₃), 3.72 (s, 6H, OCH₃), 3.66 (s, 3H, OCH₃), 3.00 (q, 2 H, NHC)H ₂ CH₂S), 2.83 (s, 2 H, COCH₂N), 2.19 (m, 16 H, NHCH₂CH ₂ S, NCH ₂ CH ₂ S, NCH ₂ CH₂CH₂CH₂CH ₂ And CH ₂ CH ₂ CO), 1.56 (m, 2 H, NCH₂CH ₂ CH₂CH₂CH₂), 1.23 (m, 4 H, NCH)₂CH₂CH ₂ CH ₂ CH₂).
¹³C NMR (CDCl₃) d 173.19, 173.14, 172.39, 172.14, 172.77 and 171.28(CO), 144.71, 144.67, 129.51, 127.87 and 126.67 (Ph), 77.45, 77.03 and 76.60 (CPh₃), 66.47 (COCH₂), 58.13 and 54.56 (NHCH), 53.81 (CH₂ CH₂CONH), 52.45 (CH₂CH₂CO), 51.76 (CH₂ CH₂CO), 37.90 (NCH₂CH₂), 36.06 (NHCH₂CH₂), 32.18 (NHCH₂ CH₂) 31.95 (NCH₂ CH₂S), 30.02 (NCH₂CH₂ CH₂CH₂CH₂), 27.90 (NCH₂ CH₂CH₂CH₂CH₂), 27.09, 26.80 and 26.61 (OCH₃), 251.15 (NCH₂CH₂CH₂CH₂ CH₂).
MS m/z 243 ((CPh₃)⁺).


[synthetic DODGA]
[7,10-Diaza-9-oxo-7-[2-(triphenylmethyl)-
Thioethyl]-12-[(triphenylmethyl)thio]-
dodecanamide-γ-L-glutamyl-L-glutamic
Acid (DODGA)]
Potassium hydroxide (2 g) was taken up in anhydrous methanol (20 mL). Adding trimethyl-γ-L-glutamate-L-glutamate-7,10-diaza-9-oxo-7-[2-((trityl)thio)ethyl] -12-[(Trityl)thio]dodedecylamine (0.15 g, 0.15 mmol) was stirred at room temperature for 30 min. Add water (10mL), adjust the pH of the reaction solution to pH = 7.0 with concentrated hydrochloric acid, extract with dichloromethane (2 × 20 mL), discard the aqueous phase, take the organic phase layer, remove the water with anhydrous sodium sulfate, reduce The product was concentrated by pressure to give the product DODGA (0.15 g, 100%).
Its analysis data:
IR (neat) n 3310 (OH), 3008 and 2927 (NH), 1741 and 1659(CO) cm^-1.
¹H NMR (CD₃OD) d 7.22 (m, 30 H, Ph), 4.42 (m, 2 H, NHCH), 3.65 (s, 2 H, COCH₂), 3.05 (t, 2 H, NHC)H ₂CH₂S), 2.86 (m, 2 H, CH ₂ CH₂CH₂CH₂CH₂), 2.77 (m, 2 H, NCH ₂ CH₂S), 2.65(m, 2 H, NCH₂CH ₂ S), 2.35 (m, 4 H, NHCH₂CH ₂ S and CH₂CH ₂ CO), 2.22 (m, 6 H, CH ₂ CH₂CONH, CH ₂ CH₂COOH and CH₂CH₂CH₂CH₂CH ₂ ), 1.99 (m, 2 H, CH₂CH ₂ COOH), 1.58 (m, 2 H, CH₂CH ₂ CH₂CH₂CH₂), 1.45 (m, 2 CH)₂CH₂CH₂CH ₂ CH₂), 1.27 (m, 2 H, CH₂CH₂CH ₂ CH₂CH₂).
¹H NMR (CD₃OD)d 175.14, 174.72, 174.58,173.83,173.79 and 173.76(CO), 144.91, 144.33, 129.58, 128.19, 127.91, 127.18 and 126.81 (Ph), 78.38 (CPh₃), 67.71 (CH), 66.77, (COCH₂), 54.03 (NHCH₂CH₂S), 51.85 (CH₂CH₂CH₂CH₂CH₂₎,38.59 (NCH₂CH₂S), 34.99 (NCH₂ CH₂S), 31.42 (NHCH₂ CH₂S), 30.18 (CH₂CH₂CO), 26.63 (CH₂ CH₂CO), 25.59 (CH₂ CH₂CH₂CH₂CH₂), 24.81 (CH)₂CH₂CH₂ CH₂CH₂), 23.30 (CH₂CH₂ CH₂CH₂CH₂).
MS m/z 243 ((CPh₃)⁺).

[Synthesis of 6'-(N-benzyloxycarbonyl)aminohexanol]
[6-(N-Benzyloxycarbonyl)aminohexanol]
6'-aminohexanol (5.9 g, 50.0 mmol) was dissolved in water (20 mL), and sodium carbonate (3.2 g, 30.0 mmol) was added and placed in an ice bath. Slowly, a solution of benzyl chlorocarbonate (7.3 g, 50.0 mmol) in diethyl ether (20 mL) was added dropwise. After the completion of the dropwise addition, the mixture was stirred at room temperature for 2 hours, and the solid was washed with a small amount of diethyl ether. The solvent was removed from the system to give the product 6'-(N-benzyloxycarbonyl)aminohexanol (9.2 g, 73.2%).
Its analysis data:
IR (neat) n 3382 and 1530 (NH), 3336 (OH), 1688 (CO) cm^-1.
¹H NMR (CDCl₃) d 7.34 (m, 5 H, Ph), 5.08 (s, 2 H, PhCH₂), 4.90 (br, 1 H, NH), 3.60 (t, J = 6.5 Hz, 2 H, CH ₂OH), 3.17 (q, J = 6.6 Hz, 2 H, NHCH ₂), 1.93 (br, 1 H, OH), 1.52 (m, 4 H, CH ₂CH₂CH₂CH ₂CH₂O), 1.35 (m, 4 H, CH ₂CH ₂CH₂CH₂O).
¹³C NMR (CDCl₃) d 156.45 (CO), 136.55, 128.42 and 127.99 (Ph), 66.51 (CH)₂OH), 62.52 (PhCH₂), 40.82 (NHCH₂), 32.45 (CH₂CH₂OH), 29.84 (NHCH₂ CH₂), 26.28 (CH₂CH₂CH₂OH), 25.22 (CH₂CH₂CH₂CH₂OH).MS m/z 251 (M⁺).

[Synthesis of 2-acetamidoamino-2,4,6-trioxo-ethenyl-1-chloro-1,2-deoxy-α-D galactopyranosole]
[2-Acetamido-3,4,6,-tri-O-acetyl-1-chloro
-1,2-dideoxy-α-D-galactopyranose]
Add acetyl chloride (30 mL) to 0 ° C, add N-acetyl-D-galactosamine (3.0 g, 13.6 mmol), cover and seal, displace Stir in a thermostat at 10 °C. After 5 days, dichloromethane (80 mL) was added and mixed, and then ice water (160 mL) was added. After stirring, the phases were separated and the organic phase was washed with saturated aqueous sodium hydrogen carbonate (1×50 mL) and then evaporated. Water, the solvent was evaporated under reduced pressure to give a viscous oily product, 2- </RTI> </RTI> </RTI> <RTIgt; </RTI> <RTIgt; </RTI> <RTIgt; </RTI> <RTIgt; Lactose (2.45 g, 51%).
Its analysis data:
IR (neat) n 3289 and 1544 (NH), 1750 and 1666 (CO) cm^-1.
¹H NMR (CDCl₃) d 6.28 (d, J = 3.6 HZ, 1 H, H₁), 5.94 (d, J = 8.7 Hz, 1 H, NH), 5.46 (dd, J = 3.2 and 1.4 Hz, 1 H, H₄), 5.29 (dd, J = 11.4 and 3.3 Hz, 1 H, H₃), 4.79 (m, 1 H, H₂), 4.48 (t, J = 6.9 Hz, 1 H, H₅), 4.19 (m, 2 H, H₆), 2.17 (s, 3 H, CH₃), 2.10 (s, 3 H, CH₃), 2.03 (s, 3 H, CH₃), 2.01 (s, 3 H, CH₃).
¹³C NMR (CDCl₃) d 170.65, 170.48, 170.26 and 169.95 (CO), 94.97 (C₁), 69.73 (C₅), 67.27 (C₄), 66.48 (C₃), 61.06 (C₂), 49.12 (C₆), 22.91, 20.56, 20.52 and 20.49 (CH)₃).
MS m/z 330 (M⁺- Cl)

[Synthesis of 6'-(N-benzyloxycarbonyl)aminohexyl-2-ethenylamino-3,4,6-trioxa-ethinyl-2-deoxy-β-D-galactoside]
[6'-(N-Benzyloxycarbonyl)aminohexyl
2-acetamido-3,4,6-tri-O-acetyl-2-deoxy-β-
D-galactopyranoside]
Take 6'-(N-benzyloxycarbonyl)aminohexanol (0.72g, 2.86mmol), 2-ethylguanidinoamino-2,4,6-trioxo-ethenyl-1-chloro-1,2-deoxy -α-D galactopyranosole (1.05 g, 2.86 mmol), anhydrous calcium sulfate (0.3 g) and mercuric cyanide (0.88 g, 3.5 mmol) co-located in toluene (15 mL) with nitromethane (15 mL), the mixture was stirred at room temperature for 24 hours, then filtered, and the filtrate was evaporated, evaporated, mjjjjjjjjjjjjjjj After concentration under reduced pressure, it was separated and purified using liquid chromatography (e.c., chloroform:methanol = 95:5) to give the colorless solid product 6'-(N-benzyloxycarbonyl)aminohexyl-2-ethylhydrazine. Amino-3,4,6-trioxa-ethinyl-2-deoxy-β-D-galactoside (0.58 g, 35 %).
Its analysis data:
IR (KBr) n 3318 and 1543 (NH), 1748 and 1668 (CO) cm^-1.
¹H NMR (CDCl₃) d 7.35 (m, 5 H, Ph), 5.94 (d, J = 8.4 Hz, NH), 5.33 (d, J = 3.0 H2, H₄), 5.26 (dd, J = 11.3 and 3.2 Hz, 1 H, H₃), 5.11 (AB, J = 12.3 Hz, 2 H, CH₂Ph), 4.90 (br, 1 H, NH), 4.65 (d, J = 8.4 Hz, 1 H, H₁), 4.12 (m, 2 H, H₆), 4.02-3.81 (m, 3 H, H₂, H₅, and OCH ₂CH₂), 3.48 (m, 1 H, OCH ₂CH₂), 3.21 (m, 2 H, CH₂N), 2.13 (s, 3 H, CH₃), 2.05 (s, 3 H, CH₃), 2.0 (s, 3 H, CH₃), 1.94 (s 3 H, CH₃), 1.51 (m, 4 H, OCH)₂CH ₂CH₂CH₂CH ₂), 1.34 (m, 4 H, OCH₂CH₂CH ₂CH ₂).
¹³C NMR (CDCl₃) d 170.44 and 156.56 (CO), 136.64, 128.49, 128.04 and 127.84 (Ph), 100.73 (C₁), 70.51, (C₅), 69.36 (C₄), 69.34 (C₃), 66.79 (CH₂Ph), 66.53 (OCH₂CH₂), 61.44 (C₆), 51.54 (C₂), 40.55 (CH₂NH), 29.70 (OCH₂ CH₂), 28.93 (NHCH₂ CH₂), 25.89 (OCH₂CH₂ CH₂), 25.07 (CH₂CH₂CH₂N), 23.35 (CH₃CONH), 20.67 (CH₃COO).
¹H NMR (CD₃OD) d 7.34 (m, 5 H, Ph), 5.32 (d, J = 3.3 Hz, 1 H, H₄), 5.05 (m, 3 H, H₃And CH₂Ph), 4.54 (d, J = 8.4 Hz, 1 H, H₁), 4.15-3.95 (m, 4 H, H₂, H₅And H₆), 3.83 (m, 1 H, OCH₂), 3.48 (m, 1 H, OCH₂), 3.10 (t, J = 6.9 Hz, 2 H, CH₂N), 2.13 (s 3 H, CH₃), 2.01 (s, 3 H, CH₃), 1.94 (s, 3 H, CH₃), 1.91 (s, 3 H, CH₃), 1.51 (m, 4 H, OCH)₂CH ₂CH₂CH₂CH ₂), 1.34 (m, 4 H, OCH₂CH₂CH ₂CH ₂).
¹³C NMR (CD₃OD) d 173.52, 172.13 and 171.11 (CO), 158.88, 138.55, 129.49 and 128.96 and 128.74 (Ph), 102.66 (C₁), 72.17 (C₅), 71.76 (C₄), 70.70 (C₃), 68.25 (CH₂Ph), 67.27 (OCH₂CH₂), 62.79 (C₆), 51.69 (C₂), 41.73 (CH₂N), 30.89 (OCH₂ CH₂), 30.51 (NHCH₂ CH₂), 27.45 (OCH₂CH₂ CH₂), 26.68 (CH₂CH₂CH₂N), 22.93 (CH₃), 20.65 (CH₃).
MS m/z 521 (M⁺- CH₃).

[synthetic ah-GalNAc₄】
[6'-Aminohexyl 2-acetamio-3,4,6-tri-O-acetyl-2-deoxy-β-D-galactopyranoside (ah-GalNAc₄)]
Take the compound 6'-(N-benzyloxycarbonyl)aminohexyl-2-ethenylamino-3,4,6-trioxa-ethinyl-2-deoxy-β-D-galactoside (0.66 g, 1.14) Methyl) was dissolved in ethanol (20 mL), 10% palladium on carbon catalyst (0.08 g) was added and placed in a hydrogenation apparatus and shaken under 50 psi of hydrogen. After about 15 to 24 hours, the filtrate was filtered, and the filtrate was evaporated to dryness to give compound ah-GalNAc.₄(0.51 g, 100%).
Its analysis data:
IR (neat) n 3256 and 3377 (NH₂), 1747 and 1657 (CO) cm^-1.
¹H NMR (CD₃OD) d 5.33 (d, J = 2.7 Hz, 1 H, H₄), 5.05 (dd, J = 11.4 and 3.3 Hz, 1 H, H₃), 4.55 (d, J = 8.4 Hz, 1 H, H₁), 4.18-3.97 (m, 4 H, H₂, H₅And H₆), 3.86 (m, 1 H, OCH ₂), 3.52 (m, 1 H, OCH ₂), 2.92 (t, J = 7.5 Hz, 2 H, CH₂N), 2.14 (s, 3 H, CH₃), 2.02 (s, 3 H, CH₃), 1.94 (s, 3 H, CH₃), 1.93 (s, 3 H, CH₃), 1.46 (m, 4 H, OCH)₂CH ₂CH₂CH₂CH ₂), 1.42 (m, 4 H, OCH₂CH₂CH ₂CH ₂).
¹³C NMR (CD₃OD) d 172.03, 171.97 and 171.61 (CO), 102.66 (C₁), 72.12 (C₅), 71.76 (C₄), 70.64 (C₃), 68.17 (OCH₂), 67.72 (C₆), 51.52 (C₂), 40.75 (CH₂N), 30.19 (OCH₂ CH₂), 28.36 (CH₂CH₂N), 27.02 (OCH₂CH₂ CH₂), 26.35 (CH₂CH₂CH₂N), 22.95 (CH₃), 20.58 (CH₃).
MS m/z 446 (M⁺), 387 (M⁺- CH₃COO).

[Synthesis of 6-trioxo-(2'-acetamido-3',4',6'-trioxo-ethenyl-2'-deoxy-β-D-galactoside)-hexyl-7,10 -diaza-9-oxo-7-[2-(trityl)thioethyl]-12-[(trityl)thio]dodecanamide-γ-L-glutamate醯-L-glutamate salt
[6-Tri-o-(2^'-acetamido-3^', 4^',6^'-
Tri-o-acetyl-2^'-deoxy-β-D-galactopyranoside)
Hexyl 7,10-diaza-9-oxo-7-[2-
(triphenylmethyl)thioethyl]-12-
[(triphenylmethyl)thio]-dodecanamido-γ-
L-glutamyl-L-glutamate]
Take compound DODGA (1.32g, 1.31mmol), compound ah-GalNAc₄(1.21 g, 5.87 mmol), triethylamine (0.82 mL, 5.87 mmol), 1,3-dicyclohexylcarbodiimide (1.21 g, 5.87 mmol) and N-hydroxybutanediamine (0.68 g, 5.87 mmol), chloroform (80 mL) was added and placed in a 250 mL round bottom flask, and heated (65 ° C) to reflux for 24 hours. Concentrated, dissolved in EA, concentrated in EA, separated and purified by liquid chromatography (cerium oxide, chloroform: methanol = 95: 5) to give the product 6-trioxo-(2'-ethylamino) -3',4',6'-trioxo-ethenyl-2'-deoxy-β-D-galactoside)-hexyl-7,10-diaza-9-oxo-7-[2 -(trityl)thioethyl]-12-[(trityl)thio]dodecanamine-γ-L-glutamate-L-glutamate (1.12 g, 37% ).
Its analysis data:
IR (neat) n 3287 and 2929 and 1538 (NH), 1748 and 1658(CO) cm^-1.
¹H NMR (CDCl₃) d7.42 (NH), 7.40-7.15 (m, 12H, Ph), 6.8-6.4 (NH), 5.35 (s, CHOAC), 5.30 (m, NHCHCO), 5.15 (m, OCHO), 4.67 ( t, CH ₂OAC), 4.14 (m, CHNHAC), 3.91 (m, CHCH₂OAC), 3.70 (m, CH₂CH ₂O), 3.45 (m, CH ₂CH₂O), 3.20 (m, NHCH ₂CH₂S), 3.00 (q, NCH ₂CH₂S), 2.83 (s, COCH₂N), 2.37 (m, NHCH₂CH ₂S and NCH₂CH ₂S), 2.22 (m, CH ₂CH₂CO), 2.05 (m, OAC), 1.54 (m, CH ₂CH ₂CH ₂ CH ₂ CH ₂CH ₂), 1.34 (m, CH₂CH₂CH ₂ CH ₂ CH₂CH₂).
MS m/z 2335 (M⁺), 2357 (M⁺Na)⁺

[synthesis (ah-GalNAc₄)₃-DODGA]
[6-Tri-o-(2^'-acetamido-3^', 4^',6^'-
Trihydroxy-2^'-deoxy-β-D-galactopyranoside)
Hexyl 7,10-diaza-9-oxo-7-[2-(triphenylmethyl)-
Thioethyl]-12-[(triphenylmethyl)thio)-
dodecanamido-γ-L-glutamyl-L-glutamate]
Take 6-trioxo-(2'-acetamido-3',4',6'-trioxo-acetamido-2'-deoxy-β-D-galactoside)-hexyl-7,10- Diaza-9-oxo-7-[2-(trityl)thioethyl]-12-[(trityl)thio]dodecanamide-γ-L-glutamate The -L-glutamate salt (1.12 g, 0.48 mmol) was dissolved in anhydrous methanol (20 mL). Slowly drip 0.1N HCl aqueous solution in an ice bath, adjust the pH to 6 and concentrate under reduced pressure to obtain the product (ah-GalNAc₄)₃-DODGA (0.94g, 100%).
Its analysis data:
IR (KBr) n 3400 (OH), 2930 and 1539 (NH), 1630 (CO) cm^-1.
¹H NMR (CD₃OD) d7.32-7.17 (m,12H, Ph) , 7.46( d, CHOH), 4.20 (m, CHCO), 3.95 (m, CHCHOH), 3.81 ( m,OCHO), 3.77 (m, COCH₂), 3.68 (d, CH₂OH), 3.53 (m, CH₂CH ₂O), 3.41 (m, CH ₂CH₂O), 3.23 (m, NHCH ₂CH₂S), 2.95 (m, NCH ₂CH₂S), 2.60 (t, NHCH ₂CH₂), 2.29 (m, CH₂CH ₂CO), 2.10 (m, CH ₂CH₂CO), 1.90 (s, NHAc), 1.45 (m, CH ₂CH ₂CH ₂ CH ₂ CH ₂CH ₂), 1.26 (m, CH₂CH₂CH ₂ CH ₂ CH₂CH₂).
MS m/z 1957 (M⁺), 1981 (M⁺Na)⁺.

The above is only the preferred embodiment of the present invention, and is not intended to limit the scope of the present invention, and the variations, modifications, and modifications of the shapes, structures, features, and spirits described in the claims of the present invention. All should be included in the scope of the patent application of the present invention.
The invention is a novelty, progressive and available for industrial use, and should meet the requirements of the patent application stipulated in the Patent Law of China, and the invention patent application is filed according to law, and the prayer bureau will grant the patent as soon as possible. prayer.

no.

First: it is the chemical structure diagram of the hepatocyte receptor marker precursor containing the trisaccharide and diazodisulfide complex in the present invention;
Figure 2 is a flow chart showing the preparation of a hepatocyte receptor label precursor containing a trisaccharide and diazodisulfide ligand in the present invention;
Third: it is a partial chemical synthesis flow chart of a preferred embodiment of the present invention;
Fourth: it is a partial chemical synthesis flow chart of a preferred embodiment of the present invention;
Figure 5 is a partial chemical synthesis flow diagram of a preferred embodiment of the present invention;
Figure 6 is a partial chemical synthesis flow diagram of a preferred embodiment of the present invention;
Figure 7 is a partial chemical synthesis flow diagram of a preferred embodiment of the present invention; and an eighth diagram: it is a hepatocyte receptor marker containing a trisaccharide and diazodisulfide ligand in the present invention. A chemical structure diagram of a contrast agent or a pharmaceutical composition of a precursor.

Claims (20)

A hepatocyte receptor marker precursor containing a trisaccharide and diazodisulfide ligand, the system is:


. A method for preparing a hepatocyte receptor label precursor containing a trisaccharide and a diazodisulfide complex, the steps of which comprise:
a carboxylic acid which synthesizes a difunctional chelating agent having a diazo disulfide ligand;
Amidating the carboxylic acid to form one of the difunctional chelating agents having a diazo disulfide ligand, the guanamine further hydrolyzed to have three polycarboxy groups;
Synthesis of three galactosides;
The galactosides are used to aminate the polycarboxyl groups of the guanamine to form a hepatocyte receptor label precursor containing a trisaccharide and a diazodisulfide ligand. The preparation method of claim 2, wherein the carboxylic acid is 7,10-diaza-9-oxo-7-[2-((trityl)thio)ethyl] -12-[(trityl)thio]dodecanoic acid. The preparation method of claim 3, wherein in the step of synthesizing the carboxylic acid, the method further comprises the steps of:
(1) using 2-thioethylamine hydrochloride and triphenylmethanol, thiol protection reaction under the catalysis of a mixture of boron trifluoride diethyl ether to form a 2-[(trityl)thio]ethylamine;
(2) using the 2-[(tritylmethyl)thio]ethylamine and chloroacetic acid chloride to carry out a guanidation reaction in a chloroform solution to form an N-[2-((tritylmethyl) group) Thio)ethyl] chloroacetamide;
(3) using the 2-[(trityl)thio]ethylamine and the N-[2-((trityl)thio)ethyl]chloroacetamide, reacted with triethylamine And carrying out a substitution reaction in a dichloromethane solvent to form a ligand containing an amine-melamine-thiol;
(4) using 6-bromohexanoic acid and sulfinium chloride, esterification reaction in an anhydrous methanol solvent to produce a methyl 6-bromohexanoate;
(5) using the amine-melamine-thiol-containing ligand and the 6-bromohexanoic acid methyl ester, using sodium hydroxide as a reactant, and performing a substitution reaction in an acetonitrile solution to form a 7,10-two Methyl aza-9-oxo-7-[2-((trityl)thio)ethyl]-12-[(trityl)thio]dodecanoate;
(6) Hydrolysis of the 7,10-diaza-9-oxo-7-[2-((trityl)thio)ethyl]-12-[(trityl)thio] Methyl diacid in an alkaline methanol solution to form the 7,10-diaza-9-oxo-7-[2-((trityl)thio)ethyl]-12-[(triphenyl) Methyl)thio]dodecanoic acid. The preparation method according to claim 2, wherein the difunctional chelating agent of the diazonium disulfide ligand of the carboxylic acid is protected with trityl group. The preparation method according to claim 4, wherein in the step (1), the reaction temperature of the thiol protection reaction is 72 ° C, and the reaction time is 4 hours. The preparation method according to the fourth aspect of the invention, wherein in the step (3), the reaction temperature of the substitution reaction is 55 ° C, and the reaction time is 48 hours. The preparation method according to the fourth aspect of the invention, wherein in the step (4), the reaction temperature of the esterification reaction is 25 ° C, and the reaction time is 24 hours. The preparation method according to the fourth aspect of the invention, wherein in the step (5), the reaction temperature of the substitution reaction is 85 ° C, and the reaction time is 24 hours. The preparation method according to claim 2, wherein the guanamine is trimethyl-γ-L-glutamate-L-glutamate-7,10-diaza-9-oxo- 7-[2-((Triphenylmethyl)thio)ethyl]-12-[(trityl)thio]dodecylamine. The preparation method of claim 10, wherein in the step of hydrating the carboxylic acid, the method further comprises the steps of:
(7) using γ-L-glutamic acid-L-glutamic acid and sulfoxide chloride to carry out esterification reaction in an anhydrous methanol solvent to form a γ-L-glutamate-L-glutamic acid trimethyl Ester;
(8) using the γ-L-glutamic acid-L-glutamic acid trimethyl ester and the carboxylic acid, and reacting with 1,3-dicyclohexylcarbodiimide and N-hydroxybutanediamine Amidoxime in a chloroform solution to form the trimethyl-γ-L-glutamate-L-glutamate-7,10-diaza-9-oxo-7-[2- ((Trityl)thio)ethyl]-12-[(trityl)thio]dodecylamine. The preparation method according to claim 11, wherein in the step (7), the reaction temperature of the esterification reaction is 25 ° C, and the reaction time is 24 hours. The preparation method according to claim 11, wherein in the step (8), the reaction temperature of the esterification reaction is 25 ° C, and the reaction time is 48 hours. The preparation method according to claim 10, wherein the guanamine is hydrolyzed to form a compound DODGA, and the structure is:


. The preparation method according to claim 2, wherein the galactoside is 6'-ethylamino-2-ethinylamino-3,4,6-trioxa-ethinyl-2-deoxy-β- D-galactoside is a compound ah-GalNAc ₄ whose structure is:


. The preparation method of claim 15, wherein in the step of synthesizing the galactoside, the method further comprises the steps of:
(9) using 6'-aminohexanol and phenyl chlorocarbonate for amino protection reaction to form a 6'-(N-benzyloxycarbonyl) aminohexanol;
(10) reacting with ethyl chlorohydrazine at 10 ° C using N-acetamidine-D-galactose to form 2-ethylamino-2,4,6-trioxo-ethenyl-1-chloro-1,2 - deoxy-α-D galactopyranosole;
(11) using the 6-(N-benzyloxycarbonyl)aminohexanol with the 2-ethylguanidino-2,4,6-trioxo-ethenyl-1-chloro-1,2-deoxy-α- D-galactopyranoside, substituted by a solution of mercury cyanide in a mixture of toluene and nitromethane to form a 6'-(N-benzyloxycarbonyl)aminohexyl-2-ethenylamino group -3,4,6-trioxo-ethenyl-2-deoxy-β-D-galactoside;
(12) using the 6'-(N-benzyloxycarbonyl)aminohexyl-2-ethenylamino-3,4,6-trioxa-ethinyl-2-deoxy-β-D-galactoside, Hydrogenation reduction reaction in ethanol under a palladium carbon catalyst to form the 6'-ethylamino-2-ethenylamino-3,4,6-trioxo-ethenyl-2-deoxy-β-D-half Glycoside. The preparation method according to claim 16, wherein in the step (9), the reaction temperature of the esterification reaction is 25 ° C, and the reaction time is 24 hours. The preparation method of claim 2, 14 or 15, wherein in the step of aminating the polyvalent carboxyl groups of the guanamine, the step further comprises the steps of:
(13) activating the carboxylic acid of the DODGA, followed by melamine in the chloroform solution with the ah-GalNAc ₄ as a reagent of 1,3-dicyclohexylcarbodiimide and N-hydroxybutanediamine. The reaction produces a 6-trioxo-(2'-acetamido-3',4',6'-trioxa-ethinyl-2'-deoxy-β-D-galactoside)-hexyl-7 ,10-diaza-9-oxo-7-[2-(trityl)thioethyl]-12-[(trityl)thio]dodecanamine-γ-L- Gluten-L-glutamate;
(14) Hydrolysis of the 6-trioxo-(2'-acetamido-3',4',6'-trioxa-ethinyl-2'-deoxy-β-D-galactoside)-hexyl- 7,10-diaza-9-oxo-7-[2-((trityl)thio)ethyl]-12-[(trityl)thio]dodecanamine-γ -L-glutamate-L-glutamate under sodium methoxide to form a 6-trioxo-(2'-acetamido-3',4',6'-trihydroxy-2'-deoxy -β-D-galactoside)-hexyl-7,10-diaza-9-oxo-7-[2-(trityl)thioethyl]-12-[(tritylmethyl) a thiol]dodecanamine-γ-L-glutamate-L-glutamate salt which is a compound (ah-GalNAc ₄ ) ₃ -DODGA. A contrast agent for a hepatocyte receptor label precursor containing a trisaccharide and a diazodisulfide complex, the structure of which is:


Wherein M is selected from the group consisting of ^99m Tc, ¹⁸⁸ Re and ¹¹¹ In. A pharmaceutical composition comprising a hepatocyte receptor marker precursor of a trisaccharide and a diazodisulfide complex, which is used for the treatment of liver cancer, comprising a compound having the following structure:


Wherein M is selected from the group consisting of ^99m Tc, ¹⁸⁸ Re and ¹¹¹ In.