阅读说明：本技术 新型糖基化二价铂抗肿瘤化合物制备方法 (Preparation method of novel glycosylated bivalent platinum anti-tumor compound ) 是由 邵佳 张强 田金磊 魏金霞 王姗 张弋 李淑娟 于 2020-12-15 设计创作，主要内容包括：本发明提供一种新型糖基化二价铂抗肿瘤化合物制备方法,其具体制备方法为：将单糖的羟基乙酰化保护后,糖环的1位被溴原子取代,得到乙酰溴代糖,随后溴原子被取代为异硫氰酸酯,再与过量水合肼反应,得到对应的乙酰化糖基氨基硫脲,进一步与不同的酮或醛反应得到目标配体。乙酰糖基化缩氨基硫脲配体在强碱条件下,得到另一系列去乙酰化糖基缩氨基硫脲目标配体。本发明的候选药物采用化学合成手段实现不同结构的糖基化配体与铂(II)连接,制备系列乙酰化/去乙酰化糖基缩氨基硫脲铂(II)目标产物,并对其进行结构表征,结构中单糖的引入,改变了铂类药物的水溶性、抗肿瘤活性及靶向性,有望为临床提供一种候选抗肿瘤药物。(The invention provides a preparation method of a novel glycosylated bivalent platinum anti-tumor compound, which comprises the following specific steps: after the hydroxyl of monosaccharide is acetylated and protected, 1 site of a saccharide ring is substituted by bromine atoms to obtain acetylbromosaccharide, then the bromine atoms are substituted by isothiocyanate, and the acetylbromosaccharide reacts with excessive hydrazine hydrate to obtain corresponding acetylated glycosyl thiosemicarbazide, and further reacts with different ketone or aldehyde to obtain the target ligand. The acetyl glycosylation thiosemicarbazone ligand obtains another series of deacetylation glycosyl thiosemicarbazone target ligand under the strong alkaline condition. The candidate drug of the invention adopts a chemical synthesis means to realize the connection of glycosylation ligands with different structures and platinum (II), prepares a series of acetylation/deacetylation glycosyl thiosemicarbazone platinum (II) target products, and carries out structural characterization on the target products, and the introduction of monosaccharide in the structure changes the water solubility, the anti-tumor activity and the targeting property of platinum drugs, and is expected to provide a candidate anti-tumor drug for clinic.)

1. A preparation method of a novel glycosylated bivalent platinum anti-tumor compound comprises the following specific steps:

1) dissolving monosaccharide in excessive acetic anhydride, reacting completely at room temperature, adding excessive HBr solution, reacting completely in a dark place, purifying, adding KSCN, refluxing to obtain acetylated glycosylated isothiocyanate, and reacting with excessive hydrazine hydrate;

2) the fully acetylated glycosylated thiosemicarbazone and different aldehydes or ketones are subjected to condensation reaction to obtain a fully acetylated glycosylated thiosemicarbazone ligand;

3) preparing a fully acetylated glycosylated thiosemicarbazone platinum (II) compound, heating and refluxing a fully acetylated glycosylated thiosemicarbazone ligand and potassium tetrachloroplatinate, and recrystallizing to obtain a pure product;

4) recrystallizing the fully acetylated glycosylated thiosemicarbazone ligand in the presence of sodium methoxide to obtain a deacetylated thiosemicarbazone ligand, heating and refluxing the deacetylated thiosemicarbazone ligand and potassium tetrachloroplatinate, recrystallizing to obtain a pure product, and synthesizing a series of glycosylated platinum (II) compounds;

5) the antitumor activity of the target glycosylation platinum (II) compound is researched, the structure-activity relationship is discussed, and the high-activity target compound is screened to research the antitumor activity.

2. The method for preparing a novel glycosylated divalent platinum anti-tumor compound according to claim 1, wherein said glycosylated platinum (II) compound has the following basic parent structure:

wherein R is₁Represents hydrogen, amino, alkyl and aromatic groups (benzene ring, pyridine ring); r₂Represents glucose, galactose, mannose, rhamnose, xylose, lyxose, ribose and acetylated glucose, acetylated galactose, acetylated mannose, acetylated rhamnose, acetylated xylose, acetylated lyxose, acetylated ribose; r₃Represents a chloride ion group.

3. The method for preparing a novel glycosylated divalent platinum antitumor compound according to claim 2, wherein the alkyl group is a methyl group, an ethyl group, an n-propyl group or an isopropyl group.

4. The method for preparing a novel glycosylated divalent platinum antitumor compound according to claim 2, wherein the aromatic group is a benzene ring or a pyridine ring.

5. The method for preparing a novel glycosylated bivalent platinum anti-tumor compound according to claim 1, wherein the dosage form of the glycosylated platinum (II) compound applied to the preparation of the anti-tumor medicament is tablets, capsules, aerosols, dispersible tablets, oral tablets, sustained release tablets, controlled release tablets, delayed release tablets, oral liquids, suppositories, dropping pills, microcapsules, microspheres, injections, freeze-dried powder injections, ointments or transdermal delivery preparations.

6. The method for preparing a novel glycosylated divalent platinum antitumor compound according to claim 1, wherein the glycosylated platinum (II) compound is a platinum (II) compound having the following different sugar structures:

the glycosylated platinum (II) compound with different structures adopts the following chemical reaction method:

1) and (3) synthesizing a fully acetylated glycosylated thiosemicarbazone ligand L-Ac: the monosaccharide is placed in acetic anhydride to react for 24-48h at room temperature, HBr replaces 1 site in a saccharide ring, bromo-sugar obtained by the reaction reacts with KSCN for 24-48h to generate acetylated saccharide isothiocyanate, and the acetylated saccharide isothiocyanate reacts with hydrazine monohydrate, wherein the feeding ratio is 1: 4-8, generating corresponding acetylated glycosyl thiosemicarbazide, and then reacting with corresponding aldehyde or ketone to obtain a corresponding fully acetylated glycosylated ligand L-Ac;

2) synthesis of deacetylated glycosylated thiosemicarbazone ligand L: reacting the fully acetylated glycosylated thiosemicarbazone ligand for 12-24h at room temperature in the presence of sodium methoxide, and recrystallizing to obtain a deacetylated glycosylated thiosemicarbazone ligand;

3) synthesis of Compounds 1-6: the platinum (II) compound needs to be prepared at the temperature of 50-100 ℃, and the feeding ratio of glycosyl thiosemicarbazone ligand to potassium tetrachloroplatinate is 1: reacting for 6-12h for 1-1.5.

7. Use of the acetylated/deacetylated glycosyl ligand and glycosylated platinum (II) compound as defined in claim 1 in the preparation of an anti-tumor medicament.

8. Use of the acetylated/deacetylated glycosyl ligand and glycosylated platinum (II) compound of claim 1 in a medicament against non-small cell lung cancer, breast cancer and cervical cancer.

Technical Field

The invention relates to the field of medical chemistry, in particular to a preparation method of a novel glycosylated bivalent platinum anti-tumor compound.

Background

Lung cancer is one of the most common cancers worldwide, with approximately 180 million new cases per year, and approximately 160 million people die of the disease. Statistical data of Chinese cancers published by professor Chenwangqing in the journal CA Cancer J Clin show that lung Cancer is also the tumor with the highest incidence and mortality in China. Lung cancer is largely divided into Small Cell Lung Cancer (SCLC) and non-small cell lung cancer (NSCLC), and more than 85% of lung cancer patients belong to NSCLC, however, NSCLC patients of about 2/3 are found to be in the middle-advanced stage, and most of these patients lose the optimal period of surgical treatment, so chemotherapy becomes one of the important means for treating NSCLC.

According to NCCN clinical practice guidelines: Non-Small Cell Lung Cancer shows that most first-line treatment regimens for NSCLC contain platinum (II) class drugs (e.g., cisplatin, carboplatin). In clinical cancer treatment, platinum (II) drugs have definite curative effects and good anti-tumor effects, and have no mutual influence with other anti-tumor drugs, so the platinum (II) drugs become one of key therapeutic drugs for a plurality of malignant solid tumors. However, the traditional platinum drugs have low selectivity and obvious toxic and side effects, and the drug resistance of tumor cells is easy to generate after long-term application, so that the targeting property of the platinum drugs needs to be improved to increase the anti-tumor activity of the platinum drugs. The "Warburg effect" is a phenomenon specific to tumor cells, and the anaerobic environment in tumors causes efficient glycolysis of cells, so that a large amount of carbohydrates such as glucose are required, thereby causing over-expression of a series of GLUT sugar receptors on the cell surface. Scholars at home and abroad actively explore glycosylated platinum drugs, synthesize a series of glucose functionalized platinum compounds, and introduce glucose into the platinum compounds to form a prodrug which shows a strong targeted GLUT1 effect. However, no literature report on glycosylated thiosemicarbazone platinum (II) compounds has been found so far.

Disclosure of Invention

The invention firstly forms the glycosylation thiosemicarbazone platinum (II) compound by the acetylation/deacetylation glycosyl thiosemicarbazone ligand and the platinum atom and the preparation method thereof, overcomes the defects of the prior art, and expects to obtain the novel glycosylation platinum compound with high water solubility, strong targeting property and anti-tumor drug resistance. According to the technical problems, the invention provides a preparation method of a novel glycosylated bivalent platinum anti-tumor compound, which comprises the following specific steps:

the monosaccharide is placed in acetic anhydride for acetylation, HBr replaces 1 position in a sugar ring to generate corresponding bromosugar, the corresponding bromosugar reacts with KSCN to generate acetylated sugar isothiocyanate, then the acetylated sugar isothiocyanate reacts with hydrazine monohydrate to generate corresponding glycosylated thiosemicarbazide, and then the corresponding glycosylated thiosemicarbazide reacts with corresponding aldehyde or ketone, including the reaction of 2-pyridylaldehyde, 2-acetylpyridine, acetophenone and 2-benzoylpyridine, so that a corresponding peracetylated ligand (L-Ac) is obtained. And heating and refluxing the prepared fully acetylated glycosyl thiosemicarbazone ligand and potassium tetrachloroplatinate in ethanol for 6-12h to obtain a corresponding compound. And reacting the fully acetylated glycosyl ligand for 12-24h at room temperature under the condition of sodium methoxide to obtain the glycosylated ligand (L). And heating and refluxing the prepared glycosylated thiosemicarbazone ligand and potassium tetrachloroplatinate in ethanol for 12-24h to obtain a corresponding glycosylated platinum (II) target product.

The glycosylated platinum (II) compound is characterized by having the following basic parent nucleus structure:

wherein R is₁Represents hydrogen, amino, alkyl (methyl, ethyl, n-propyl, isopropyl) and aromatic groups (benzene, pyridine rings); r₂Represents glucose, galactose, mannose, rhamnose, xylose, lyxose, ribose and peracetylatedGlucose, acetylated galactose, acetylated mannose, acetylated rhamnose, acetylated xylose, acetylated lyxose, acetylated ribose; r₃Represents a chloride ion group.

The glycosylated platinum (II) compound is applied to the preparation of antitumor drugs, and the administration dosage form of the glycosylated platinum (II) compound is tablets, capsules, aerosols, dispersible tablets, oral tablets, sustained-release tablets, controlled-release tablets, delayed-release tablets, oral liquid, suppositories, dropping pills, microcapsules, microspheres, injections, freeze-dried powder injections, ointments or transdermal administration preparations.

The glycosylated platinum (II) compound is a glycosylated platinum (II) compound containing the following different sugar structures:

the chemical reaction method of the glycosylated platinum (II) compounds with different structures is as follows (the following synthetic routes and methods are all described by taking the structures of representative 6 glycosylated platinum (II) compounds as examples, the synthesis or preparation method of the compounds with similar reaction principles to the compounds can refer to the routes or methods, and the specific groups or experimental conditions of the synthesis can be adjusted according to the actual conditions):

1) and (3) synthesizing a fully acetylated glycosylated thiosemicarbazone ligand L-Ac: monosaccharide is placed in acetic anhydride to react at room temperature for 12-24h at a feeding ratio of 1: 6-10, HBr replaces 1 position in a sugar ring at a feeding ratio of 1: 2-5, bromo-sugar obtained by the reaction reacts with KSCN at 24-48h at a feeding ratio of 1: 2-3 to generate acetylated sugar isothiocyanate, and the acetylated sugar isothiocyanate reacts with hydrazine monohydrate at a feeding ratio of 1: 4-8, generating corresponding acetylated glycosyl thiosemicarbazide, and then reacting with corresponding aldehyde or ketone with the feeding ratio of 1: 1-1.5, and obtaining a corresponding fully acetylated glycosylated ligand L-Ac; some of the intermediate synthesis methods are described in the references: [1] Alia-Cristina Tencishi (Deleanu), Ionanis D.Kostas, Dimitra Kovala-Demertzi, et al, Synthesis and characterization of new aromatic aldehyde/ketone 4- (b-D-glucopyranosyl) thiosemiconductor, Carbohydrate Research,2009,344: 1352. 1364.[2] Kyra-Melinda Alexaacuo, Alia-Cristina Tencishi (Deleanu), Evanlia D.Chrysysina, The polarization of b-D-glucopyranosyl-thiosemiconductor modification to phosphorus enzyme, A. simulation A. restriction of biological, 9. Biochemical of chemical, 9. Biochemical, 11. Biochemical, et al.

2) Synthesis of deacetylated glycosylated thiosemicarbazone ligand L: the fully acetylated glycosylated thiosemicarbazone ligand is added in a ratio of 1: 6-10 in the presence of sodium methoxide, the mixture reacts for 12-24 hours at room temperature, and the deacetylated glycosylated thiosemicarbazone ligand is obtained through recrystallization;

3) synthesis of Compounds 1-6: the platinum (II) compound needs to be prepared at the temperature of 50-100 ℃, and the feeding ratio of glycosyl thiosemicarbazone ligand to potassium tetrachloroplatinate is 1: reacting for 6-12h for 1-1.5.

The invention also aims to provide the application of the acetylated/deacetylated glycosyl ligand and the glycosylated platinum (II) compound in preparing antitumor drugs.

The invention also aims to provide the application of the acetylated/deacetylated glycosyl ligand and the glycosylated platinum (II) compound in medicines for resisting non-small cell lung cancer, breast cancer and cervical cancer.

The candidate drug of the invention realizes the connection of glycosylation ligands with different structures and platinum (II) by a chemical synthesis means, synthesizes a series of glycosylation platinum (II) target products, researches the antitumor activity of a target compound, discusses the structure-activity relationship, screens out a high-activity target compound and researches the antitumor activity mechanism of the high-activity target compound. The introduction of monosaccharide changes the water solubility, the anti-tumor activity and the targeting property of platinum drugs, the invention carries out original innovative researches on the design, the synthesis, the anti-cancer activity, the action mechanism and the like of a series of glycosylated bivalent platinum anti-tumor compounds, is expected to discover a novel anti-tumor compound with broad spectrum, high efficiency and low toxicity and independent intellectual property rights, and provides a novel candidate drug for clinical treatment of tumors. The invention of the innovative medicine from the source has important theoretical value and practical significance for national economy, social development, people's health and the like.

A preparation method of a novel glycosylated bivalent platinum anti-tumor compound comprises the following specific steps:

the terms used in the present invention have meanings conventionally understood by those skilled in the art, and some of the terms used herein are defined as follows in order to facilitate understanding of the present invention:

the glycosyl platinum (II) compound referred to in the invention is not particularly specified, but refers to an acetylated glycosyl platinum (II) compound and a deacetylated glycosyl platinum (II) compound; the glycosylated ligands are not specifically described, but both of the acetylated glycosyl ligands and the deacetylated glycosyl ligands are referred to. L represents a deacetylated glycosylated thiosemicarbazone ligand; L-Ac represents a fully acetylated glycosylated thiosemicarbazone ligand; gal represents galactose, glu represents glucose; l is₁-gal～L₈-gal represents an intermediate in the synthesis of the galactosylated thiosemicarbazone ligand; l is₁-glu～L₆-glu represents an intermediate in the synthesis of a glucosylated thiosemicarbazone ligand; a. the₁～A₄Represents a galactose platinum (II) compound, B₁～B₄Represents a platinum (II) glucose compound. CDDP represents the positive control cisplatin.

The different glycoplatinum (II) compounds of the present invention are prepared by conventional techniques known to those skilled in the art or by commercially available reagents, all of which are mentioned herein, except where specifically indicated, from high purity reagents that the reagent company meets the experimental requirements.

As used in the specification and in the claims, the singular form of "a", "an", and "the" include plural references unless the context clearly dictates otherwise. All numerical designations such as temperature, time, concentration, including ranges, are approximations. The description herein is merely exemplary and equivalents thereof known in the art.

1) Dissolving monosaccharide in excessive acetic anhydride at a feeding ratio of 1: 6-10, adding excessive HBr solution at a feeding ratio of 1: 2-5 after complete reaction at room temperature, purifying after complete reaction in a dark place, adding KSCN at a reflux feeding ratio of 1: 2-3 to obtain fully acetylated glycosylated isothiocyanate, and reacting with excessive hydrazine hydrate at a feeding ratio of 1: 4-8; partial intermediate synthesis methods are described in references: [1] Alia-Cristina Tencishi (Deleanu), Ionanis D.Kostas, Dimitra Kovala-Demertzi, et al, Synthesis and characterization of new aromatic aldehyde/ketone 4- (b-D-glucopyranosyl) thiosemiconductor, Carbohydrate Research,2009,344: 1352. 1364.[2] Kyra-Melinda Alexaacuo, Alia-Cristina Tencishi (Deleanu), Evanlia D.Chrysysina, The polarization of b-D-glucopyranosyl-thiosemiconductor modification to phosphorus enzyme, A. simulation A. restriction of biological, 9. Biochemical of chemical, 9. Biochemical, 11. Biochemical, et al.

2) The ratio of the total acetylated and saccharified thiosemicarbazide to different aldehydes or ketones for condensation reaction is 1: 1-1.5, obtaining a fully acetylated glycosylated thiosemicarbazone ligand; synthetic methods are described in references: Zhong-Ying Ma, Jia Shao, Wei-Guo Bao, et al, a thiosemiconductor baseband chip (II) complex as a potential anti agent, Journal of coding Chemistry 2015,68,2: 277-.

3) Preparing a fully acetylated glycosylated thiosemicarbazone platinum (II) compound, wherein the feeding ratio of a fully acetylated glycosylated thiosemicarbazone ligand to potassium tetrachloroplatinate is 1: 1-1.5, heating and refluxing ethanol, and recrystallizing to obtain a pure product; synthetic methods are described in references: jia Shao, Wei-Guo Bao, He Tian, et al, nucleic activity and protein-binding properties of a novel thiosemiconductor Pt (II) complex, Dalton trans, 2014,43: 1663-.

4) The peracetylated glycosylated thiosemicarbazone ligand was reacted in the presence of sodium methoxide 1: 6-10, and recrystallizing to obtain the deacetylated glycosyl thiosemicarbazone ligand. The material ratio of the deacetylated glycosyl thiosemicarbazone ligand to potassium tetrachloroplatinate is 1: 1-1.5, heating and refluxing ethanol, recrystallizing to obtain a pure product, and synthesizing a series of glycosylated platinum (II) compounds;

5) the antitumor activity of the target glycosylation platinum (II) compound is researched, the target compound with high antitumor activity is screened out, and the structure-activity relationship is discussed.

The invention has the beneficial effects that: the candidate drug of the invention realizes the connection of glycosylated ligands with different sugar structures and platinum (II) by a chemical synthesis means, and synthesizes a series of glycosylated platinum (II) target products. The invention designs and synthesizes a class of acetylated/deacetylated glycosyl thiosemicarbazone platinum (II) compounds, performs structural characterization on the compounds, and measures the corresponding antitumor activity so as to obtain new platinum (II) medicines with high water solubility, strong targeting property and antitumor drug resistance. The introduction of monosaccharide changes the water solubility, the anti-tumor activity and the targeting property of platinum drugs.

The invention relates to a preparation method of a novel glycosylated platinum (II) compound and an antitumor activity test thereof. The glycosylated platinum (II) medicine is not clinically applied at present, but part of known glycosylated platinum compounds have strong advantages in the aspects of water solubility, targeting and anti-tumor drug resistance. The invention creatively prepares a glycosylation platinum (II) compound which is not reported in the literature, the antitumor activity of the compound in specific cells is superior to that of cisplatin, and in addition, the glycosylation ligand can endow the platinum compound with higher targeting property to tumor cells, and the selectivity of the medicament to the tumor cells can be improved. Meanwhile, the glycosyl ligand is introduced, so that the water solubility of the platinum drugs is effectively improved, and various adverse reactions generated in the chemotherapy process of the platinum drugs can be effectively reduced. The invention is expected to find a novel broad-spectrum, high-efficiency and low-toxicity anti-tumor compound with independent intellectual property rights, and provides a novel candidate active molecule for clinical tumor treatment. The invention of innovative medicaments at the source has great significance for further development of human beings in defeating cancer and national economy.

Drawings

The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate embodiments of the invention and, together with the description, serve to explain the invention without limitation.

FIG. 1 is a scheme showing the synthesis of the peracetylated glycosylated thiosemicarbazone ligand L-Ac.

FIG. 2 is a scheme showing the synthesis of a deacetylated glycosylated thiosemicarbazone ligand L.

FIG. 3 is a general scheme for the synthesis of platinum (II) compounds 1-6.

FIG. 4 is the mother nucleus structure of glycosylated platinum compound in the invention.

FIG. 5 is a graph showing the survival rate of A549 cells relative to the concentration of glycosylated platinum compound in the invention.

FIG. 6 is a graph showing the concentration of glycosylated platinum compound and the survival rate of H460 cells in the invention.

FIG. 7 is a graph showing the concentration of glycosylated platinum compound and the survival rate of MCF7 cells in the invention.

FIG. 8 is a graph showing the survival rate of Hela cells in the invention with respect to the concentration of glycosylated platinum compound.

Detailed Description

As used in the specification and in the claims, the singular form of "a", "an", and "the" include plural references unless the context clearly dictates otherwise. All numerical designations such as temperature, time, concentration, including ranges, are approximations. The description herein is merely exemplary and equivalents thereof known in the art.

The glycosyl platinum (II) compound referred to in the invention is not particularly specified, but refers to an acetylated glycosyl platinum (II) compound and a deacetylated glycosyl platinum (II) compound; the glycosylated ligands are not specifically described, but both of the acetylated glycosyl ligands and the deacetylated glycosyl ligands are referred to. L represents a deacetylated glycosylated thiosemicarbazone ligand; L-Ac represents a fully acetylated glycosylated thiosemicarbazone ligand; gal represents galactose, glu represents glucose; l is₁-gal～L₈-gal represents an intermediate in the preparation of the galactosylated thiosemicarbazone ligand; l is₁-glu～L₆-glu represents an intermediate in the preparation of a glucosylated thiosemicarbazone ligand; a. the₁～A₄Represents a galactose platinum (II) complex, B₁～B₄Represents a platinum (II) glucose complex. CDDP represents the positive control cisplatin.

The different glycoplatinum (II) compounds of the present invention are prepared by conventional techniques known to those skilled in the art or as off-the-shelf reagents, and the various reagents mentioned herein are obtained from reagent companies, unless otherwise specified, as high purity reagents meeting the experimental requirements.

Example 1

The invention provides a preparation method of a novel glycosylated bivalent platinum anti-tumor compound, which comprises the following steps:

the preparation method of the glycosylated platinum (II) compound comprises the steps of placing monosaccharide in acetic anhydride for acetylation, substituting HBr for 1 position in a sugar ring to generate corresponding bromosugar, reacting with KSCN to generate acetylated sugar isothiocyanate, then reacting with hydrazine monohydrate to generate corresponding glycosyl thiosemicarbazide, and then reacting with corresponding aldehyde or ketone, including the reaction of 2-pyridylaldehyde, 2-acetylpyridine, acetophenone, 2-benzoylpyridine and the like to obtain the corresponding peracetylated glycosylated ligand (L-Ac). And heating and refluxing the prepared fully acetylated glycosyl thiosemicarbazone ligand and potassium tetrachloroplatinate in ethanol for 6-12h to obtain a corresponding glycosylated platinum (II) compound. And recrystallizing the fully acetylated glycosylated thiosemicarbazone ligand in the presence of sodium methoxide to obtain the deacetylated glycosyl thiosemicarbazone ligand. And (3) heating and refluxing the deacetylated glycosyl thiosemicarbazone ligand, potassium tetrachloroplatinate and ethanol for 6-12h, and recrystallizing to obtain a pure product to obtain a series of glycosylated platinum (II) compounds. The antitumor activity of the target glycosylation platinum (II) compound is researched, the target compound with high antitumor activity is screened out, and the structure-activity relationship is discussed. .

Example 2

The glycosylated platinum (II) compound has the following basic parent nucleus structure:

wherein R is₁Represents hydrogen, amino, alkyl (methyl, ethyl, n-propyl, isopropyl) and aromatic groups (benzene, pyridine rings); r₂Represents glucose, galactose, mannose, rhamnose, xylose, lyxose, ribose and acetylated glucose, acetylated galactose, acetylated mannose, acetylated rhamnose, acetylated xylose, acetylated lyxose, acetylated ribose; r₃Represents a chloride ion group.

Representative compounds of glycosylplatinum (II) compounds provided by the present invention can be provided in table 1 below, but are not limited to the following compounds:

table 1 Structure of the Compounds

Example 3

The preparation method of the glycosylated thiosemicarbazone ligand comprises the following steps:

taking galactose as an example, a complete synthetic route for one embodiment of the glycosyl ligand is as follows:

the feed ratio of acetylated galactose in acetic anhydride is 1: 6-10, the feed ratio of 1 site in HBr bromo-sugar ring is 1: 2-5, bromo-acetylgalactosamine is generated, the bromo-acetylgalactosamine further reacts with KSCN to generate acetylgalactosamine isothiocyanate with the feed ratio of 1: 2-3, and the reaction feed ratio of the bromo-acetylgalactosamine to hydrazine monohydrate is 1: 4-8, producing the corresponding 4-acetylgalactosaminthiosemicarbazide. Reacting 4-acetylgalactosaminthiosemicarbazide with corresponding aldehyde or ketone, wherein the reaction comprises the reaction of 2-pyridylaldehyde, 2-acetylpyridine, acetophenone and 2-benzoylpyridine (the feeding ratio is 1: 1-1.5), and obtaining the corresponding fully acetylated glycosylated ligand. The feeding ratio of the fully acetylated glycosylated thiosemicarbazone ligand in the presence of sodium methoxide is 1: and 6-10, recrystallizing to obtain the glycosylated thiosemicarbazone ligand.

L₂-gal：¹H NMR(400MHz,Chloroform-d):δ6.63(d,J＝3.9Hz,1H),5.45(dd,J＝3.3,1.4Hz,1H),5.34(dd,J＝10.7,3.3Hz,1H),4.98(dd,J＝10.7,4.0Hz,1H),4.42(td,J＝6.5,1.3Hz,1H),4.12(dd,J＝11.4,6.3Hz,1H),4.07–4.01(m,1H),2.09(s,3H),2.05(s,3H),2.00(s,3H),1.95(s,3H).¹³C NMR(400MHz,Chloroform-d):δ170.33,170.08,169.91,169.77,88.12,71.05,67.98,67.76,66.96,60.83,20.77,20.66,20.61,20.58。

L₃-gal：¹H NMR(400MHz,Chloroform-d)δ5.44(d,J＝3.3Hz,1H),5.36(dd,J＝10.3,8.7Hz,1H),5.11–4.99(m,2H),4.17(d,J＝6.5Hz,2H),4.01(t,J＝6.5Hz,1H),2.21(s,3H),2.16(s,3H),2.09(s,4H),2.03(s,3H).¹³C NMR(101MHz,Chloroform-d)δ170.37,170.09,169.96,169.17,143.75,84.00,72.96,70.62,69.29,66.87,61.30,61.27,61.24,20.70,20.65,20.55。

L₄-gal：¹H NMR(400MHz,Chloroform-d)δ8.14(d,J＝9.3Hz,1H),7.76(s,1H),5.67(t,J＝9.1Hz,1H),5.40(d,J＝3.2Hz,1H),5.30–5.06(m,2H),4.09(dt,J＝5.9,3.4Hz,2H),4.02(q,J＝6.6,5.2Hz,1H),3.79(s,2H),2.10(s,3H),2.00(d,J＝6.1Hz,6H),1.94(s,3H).¹³C NMR(101MHz,Chloroform-d)δ183.77,171.18,169.82,82.48,72.44,70.95,68.48,67.34,61.23,20.90,20.79,20.69,20.61。

L₅-gal：¹H NMR(400MHz,Chloroform-d)δ8.95(s,1H),8.61(dt,J＝4.8,1.4Hz,1H),8.47(dd,J＝16.0,9.0Hz,1H),8.25(d,J＝8.0Hz,1H),7.78(td,J＝7.8,1.8Hz,1H),7.32(ddd,J＝7.5,4.8,1.2Hz,1H),5.73(t,J＝9.0Hz,1H),5.55–5.46(m,1H),5.36(td,J＝9.8,9.2,1.8Hz,1H),5.25(td,J＝9.8,3.4Hz,1H),4.21–4.01(s,3H),2.43(s,J＝2.1Hz,3H),2.18(s,J＝2.9Hz,3H),2.06(s,J＝4.8Hz,6H),2.02(s,3H).¹³C NMR(101MHz,Chloroform-d)δ179.63,171.45,170.46,170.12,169.70,154.16,149.04,148.69,136.47,124.26,120.96,82.68,72.39,70.74,68.34,67.29,61.08,20.83,20.72,20.67,20.54,11.33。

L₆-gal：¹H NMR(400MHz,DMSO-d₆)δ10.71(s,1H),8.61(d,J＝4.8Hz,1H),8.51(d,J＝9.2Hz,1H),8.29(d,J＝8.1Hz,1H),7.86(td,J＝7.7,1.8Hz,1H),7.43(dd,J＝7.3,5.0Hz,1H),5.35(t,J＝9.0Hz,1H),5.02(d,J＝5.6Hz,1H),4.83(d,J＝5.3Hz,1H),4.62(t,J＝5.4Hz,1H),4.39(d,J＝5.4Hz,1H),3.72(dt,J＝10.3,4.6Hz,2H),3.54(qd,J＝8.8,5.5Hz,1H),3.44(dq,J＝9.3,5.5Hz,3H),2.43(s,3H).¹³C NMR(101MHz,DMSO-d₆)δ179.63,154.48,149.27,148.62,136.56,124.17,120.79,84.37,76.80,74.01,69.58,68.19,60.12,12.52。

L₇-gal:¹H NMR(400MHz,Chloroform-d)δ9.95(s,1H),8.67(d,J＝4.7Hz,1H),8.47(d,J＝8.6Hz,1H),8.17(d,J＝8.0Hz,1H),7.99(s,1H),7.83(td,J＝7.8,1.8Hz,1H),7.37(dd,J＝7.3,5.0Hz,1H),5.74(t,J＝8.9Hz,1H),5.54(d,J＝3.4Hz,1H),5.40(t,J＝9.7Hz,1H),5.29(dd,J＝10.3,3.4Hz,1H),4.22(d,J＝5.8Hz,2H),4.19–4.13(m,1H),2.22(s,3H),2.10(d,J＝2.4Hz,6H),2.06(s,3H).¹³C NMR(101MHz,Chloroform-d)δ179.32,171.54,170.54,170.18,169.79,152.21,149.48,143.50,136.90,124.66,120.96,82.63,72.39,70.74,68.36,67.27,61.09,20.86,20.78,20.70,20.59。

L₈-gal：¹H NMR(400MHz,DMSO-d₆)δ11.98(s,1H),8.68(d,J＝9.1Hz,1H),8.63–8.57(m,1H),8.33(d,J＝8.0Hz,1H),8.18(s,1H),7.87(td,J＝7.7,1.7Hz,1H),7.42(ddd,J＝7.5,4.9,1.2Hz,1H),5.35(t,J＝9.0Hz,1H),4.94(s,2H),4.66(s,1H),4.47(s,1H),3.88–3.67(m,2H),3.61–3.43(m,4H).¹³C NMR(101MHz,DMSO-d₆)δ179.36,149.94,143.75,137.08,124.82,121.05,85.01,77.38,74.69,69.91,68.72,60.72。

Taking glucose as an example, a complete synthetic route for one embodiment of glycosyl ligands is as follows:

the above synthetic routes are not exclusive, and the specific reaction conditions can be checked according to the published documents in the prior art, and other reaction routes capable of obtaining the target reactant can be implemented.

And (3) synthesizing a glucose thiosemicarbazone ligand: reacting bromoacetylated glucose with KSCN at a feed ratio of 1: 2-3, refluxing for 24-48h to generate acetyl glucose isothiocyanate, and reacting according to the weight ratio of 1: adding hydrazine monohydrate in a ratio of 4-8, and stirring at room temperature for 48 hours to generate the corresponding 4-acetyl glucosyl thiosemicarbazide. Reacting 4-acetyl glucosyl thiosemicarbazide with corresponding aldehyde or ketone, wherein the reaction comprises the reaction of 2-pyridylaldehyde, 2-acetylpyridine and 2-benzoylpyridine (the feeding ratio is 1: 1-1.5), and obtaining a corresponding three-coordination glycosylation ligand. The peracetylated glycosylated thiosemicarbazone ligand was reacted in the presence of sodium methoxide 1: and 6-10, recrystallizing to obtain the glycosylated thiosemicarbazone ligand.

L₁-glu：¹H NMR(400MHz,Chloroform-d)δ5.15(td,J＝9.4,4.0Hz,1H),5.05(td,J＝9.9,9.4,4.3Hz,2H),4.96(dd,J＝8.5,4.4Hz,1H),4.17(dd,J＝10.9,5.9Hz,1H),4.14–4.01(m,1H),3.68(d,J＝9.1Hz,1H),2.10–1.86(m,12H).¹³C NMR(101MHz,Chloroform-d)δ169.54,169.07,168.20,168.03,143.20,82.46,73.03,71.46,70.85,66.62,60.49,19.68,19.51。

L₂-glu：¹H NMR(400MHz,Chloroform-d)δ8.08(d,J＝9.2Hz,1H),7.66(s,1H),5.69(t,J＝8.5Hz,1H),5.31(td,J＝9.5,2.6Hz,1H),5.03(tdd,J＝9.8,7.2,2.6Hz,2H),4.32–4.19(m,1H),4.09–4.01(m,1H),3.80(ddd,J＝9.8,4.9,2.5Hz,3H),2.05–1.92(m,12H).¹³C NMR(101MHz,Chloroform-d)δ183.70,170.86,170.69,169.88,169.61,82.07,73.50,72.75,70.64,68.26,61.61,20.77,20.61。

L₃-glu：¹H NMR(400MHz,Chloroform-d)δ8.92(s,1H),8.64(d,J＝4.8Hz,1H),8.53(s,1H),8.24(d,J＝8.0Hz,1H),7.82(t,J＝7.8Hz,1H),7.43–7.32(m,1H),5.77(t,J＝9.0Hz,1H),5.44(q,J＝9.8Hz,1H),5.19(dt,J＝25.3,9.7Hz,2H),4.40(dd,J＝12.5,4.6Hz,1H),4.16(dd,J＝12.4,2.2Hz,1H),3.95(ddd,J＝10.3,4.7,2.2Hz,1H),2.44(s,3H),2.15–1.88(m,12H).¹³C NMR(101MHz,Chloroform-d)δ178.64,170.14,169.65,168.82,168.59,147.55,146.95,135.70,123.30,122.89,119.89,81.37,72.56,71.64,69.57,67.37,60.63,19.76,19.72,19.60,10.25。

L₄-glu：¹H NMR(400MHz,DMSO-d₆)δ10.70(s,1H),8.61(q,J＝6.1Hz,2H),8.43–8.31(m,1H),7.85(p,J＝7.6Hz,1H),7.43(ddd,J＝15.3,9.7,5.5Hz,1H),5.43(p,J＝8.8Hz,1H),5.07(dt,J＝15.0,7.4Hz,2H),4.94(q,J＝5.2Hz,1H),4.50(dt,J＝11.4,5.6Hz,1H),4.11(dq,J＝10.5,5.3Hz,1H),3.66(p,J＝6.4Hz,1H),3.39–3.31(m,1H).¹³C NMR(101MHz,DMSO-d₆)δ180.22,154.97,149.77,149.05,136.96,124.64,121.51,84.51,79.14,77.98,72.53,70.28,61.25,12.94。

L₅-glu：¹H NMR(400MHz,Chloroform-d)δ14.72(s,1H),8.77(d,J＝4.9Hz,1H),8.40(d,J＝9.3Hz,1H),7.91(td,J＝7.8,1.7Hz,1H),7.48(d,J＝7.9Hz,1H),7.42(dd,J＝7.6,5.0Hz,1H),7.22(s,1H),5.93(s,1H),5.44(d,J＝9.4Hz,1H),5.26–5.11(m,2H),4.40(dd,J＝12.5,4.5Hz,1H),4.18(d,J＝12.5Hz,1H),4.00–3.91(m,1H),2.14(s,3H),2.09(t,J＝2.2Hz,10H).¹³C NMR(101MHz,Chloroform-d)δ180.12,170.73,169.94,169.66,151.77,148.58,137.61,134.30,125.83,124.35,82.09,73.65,72.98,70.66,68.39,61.78,20.82,20.72,20.67。

L₆-glu：¹H NMR(400MHz,DMSO-d₆)δ11.02(s,1H),9.09(s,1H),8.85(d,J＝5.2Hz,1H),8.39(d,J＝3.8Hz,2H),7.85(s,1H),5.37(t,J＝8.9Hz,1H),3.86(d,J＝2.7Hz,1H),3.84(d,J＝2.7Hz,1H),3.81(d,J＝2.7Hz,1H),3.77(s,1H),3.57(dd,J＝12.1,8.7Hz,2H),3.49–3.41(m,4H).¹³C NMR(101MHz,DMSO-d₆)δ179.44,153.41,149.74,143.56,137.22,124.84,121.31,84.59,79.18,78.01,72.33,70.23,61.23。

Example 4

Synthesis of glycosylated platinum (II) compounds

Taking galactose as an example, an embodiment of a galactoplatinum (II) compound is fully synthesized as follows:

respectively heating galactosylation ligand and potassium tetrachloroplatinate in ethanol for 12-24h, wherein the feeding ratio is 1: 1-1.5, obtaining the corresponding galactosylation platinum compound, and recrystallizing to obtain a pure product.

A₁:¹H NMR(400MHz,Chloroform-d)δ8.69(s,1H),7.90(t,J＝7.9Hz,1H),7.40(d,J＝8.0Hz,1H),7.34(s,1H),5.46(s,1H),5.18(d,J＝7.2Hz,3H),4.21(d,J＝6.5Hz,2H),4.04(t,J＝6.7Hz,1H),2.37(s,3H),2.17(s,3H),2.05(d,J＝11.7Hz,7H),1.97(s,2H).¹³C NMR(101MHz,DMSO-d₆)δ170.50,170.13,169.82,169.40,146.76,141.21,127.87,126.64,83.34,73.56,72.56,71.33,68.31,62.20,21.07,20.99,20.89,20.85,13.88.C₂₂H₂₇ClN₄O₉PtS(M+H⁺) Theoretical value 755.0790, test value 755.0893.

A₂:¹H NMR(400MHz,DMSO-d₆)δ9.20(s,1H),8.78(d,J＝5.7Hz,1H),8.52(s,1H),8.24(t,J＝7.8Hz,1H),7.85(d,J＝7.9Hz,1H),7.78(t,J＝6.8Hz,1H),5.49(s,1H),5.41–5.28(m,2H),5.19(t,J＝9.4Hz,1H),4.37(s,1H),4.09(d,J＝6.8Hz,2H),2.19(s,3H),2.06(d,J＝2.4Hz,6H),1.98(s,3H).¹³C NMR(101MHz,DMSO-d₆)δ170.35,169.88,169.37,159.65,147.35,141.46,127.32,126.77,83.76,71.90,71.42,69.16,67.93,61.80,21.01,20.96,20.84。C₂₁H₂₅ClN₄O₉PtS(M+H⁺) Theoretical value 741.0520, test value 741.0728.

A₃:¹H NMR(400MHz,Chloroform-d)δ8.69(s,1H),7.90(t,J＝7.9Hz,1H),7.40(d,J＝8.0Hz,1H),7.34(s,1H),5.46(s,1H),5.18(d,J＝7.2Hz,3H),4.21(d,J＝6.5Hz,2H),4.04(t,J＝6.7Hz,1H),2.37(s,3H),2.17(s,3H),2.05(d,J＝11.7Hz,7H),1.97(s,2H).¹³C NMR(101MHz,DMSO-d₆)δ182.09,159.97,146.46,141.07,127.47,126.20,86.91,77.40,74.67,70.11,68.68,60.89,13.74。C₁₄H₁₉ClN₄O₅PtS(M+H⁺) Theoretical value 586.9310, test value 587.0010.

A₄:¹H NMR(400MHz,DMSO-d₆)δ8.86(d,J＝8.9Hz,1H),8.72(d,J＝5.8Hz,1H),8.46(s,1H),8.16(t,J＝7.9Hz,1H),7.74(d,J＝8.0Hz,1H),7.70(d,J＝7.0Hz,1H),4.83(dd,J＝20.9,12.1Hz,3H),4.64(s,1H),4.47(s,1H),3.68(s,1H),3.48(dt,J＝16.8,10.3Hz,5H).¹³C NMR(101MHz,DMSO-d₆)δ184.42,159.73,149.18,147.12,141.33,126.97,126.33,86.71,77.32,74.58,70.13,68.50,60.80。C₁₃H₁₇ClN₄O₅PtS(M+H⁺) Theoretical value 572.9040, test value 573.0299.

A complete synthetic route to an embodiment of a glycosylplatinum (II) compound, exemplified by glucose, is the following:

and respectively heating the glucose glycosylation ligand and potassium tetrachloroplatinate in ethanol for 12-24h, wherein the feeding ratio is 1: 1-1.5, obtaining a corresponding glucosyl platinum compound, and recrystallizing to obtain a pure product.

B₁:¹H NMR(400MHz,Chloroform-d)δ8.76–8.54(m,1H),7.87(t,J＝7.8Hz,1H),7.46–7.24(m,2H),6.26(s,1H),5.33(t,J＝9.4Hz,1H),5.18(t,J＝9.3Hz,1H),5.05(t,J＝9.7Hz,1H),4.96(t,J＝9.2Hz,1H),4.25(dd,J＝12.5,5.1Hz,1H),4.18–4.07(m,1H),3.78(d,J＝7.9Hz,1H),2.30(s,3H),2.01(dd,J＝19.8,6.3Hz,12H).¹³C NMR(101MHz,Chloroform-d)δ169.95,169.58,168.95,168.61,159.41,145.56,138.99,125.30,124.78,83.06,72.65,71.63,69.86,67.56,61.04,59.38,19.83,19.79,19.59,13.19,13.01。C₂₂H₂₇ClN₄O₉PtS(M+H⁺) Theoretical value 755.0790, test value 755.0893.

B₂:¹H NMR(400MHz,Chloroform-d)δ8.65(s,1H),7.93–7.78(m,2H),7.42(d,J＝7.8Hz,1H),7.35(d,J＝6.6Hz,1H),6.65(s,1H),5.30(t,J＝9.5Hz,1H),5.18(t,J＝9.3Hz,1H),5.04(t,J＝9.7Hz,1H),4.97(d,J＝9.7Hz,1H),4.26(dd,J＝12.5,4.8Hz,1H),4.10(dd,J＝12.6,2.3Hz,1H),3.85–3.72(m,1H),2.04(d,J＝3.8Hz,6H),1.98(d,J＝5.7Hz,6H).¹³C NMR(101MHz,DMSO-d₆)δ170.44,170.01,169.75,169.33,159.63,147.37,141.49,127.38,126.85,83.20,73.31,72.50,71.47,68.12,62.08,21.02,20.88,20.85,20.79。C₂₁H₂₅ClN₄O₉PtS(M+H⁺) Theoretical value 741.0520, test value 741.0745.

B₃:¹H NMR(400MHz,DMSO-d₆)δ9.05(s,1H),8.79(d,J＝5.5Hz,4H),8.62(s,3H),8.19(t,J＝7.9Hz,4H),7.89–7.58(m,9H),4.87(s,3H),4.07(s,38H),3.66(d,J＝11.8Hz,4H),3.45(dd,J＝12.3,4.8Hz,4H),3.32–3.00(m,19H),2.33(s,13H).¹³C NMR(101MHz,DMSO-d₆)δ182.11,160.03,157.50,146.61,141.15,127.62,126.25,86.46,79.37,78.13,72.82,70.33,61.27,13.76。C₁₄H₁₉ClN₄O₅PtS(M+H⁺) Theoretical value 586.9310, test value 587.0460.

B₄:¹H NMR(400MHz,DMSO-d₆)δ8.86(d,J＝8.8Hz,1H),8.73(s,1H),8.51(s,1H),8.18(t,J＝7.8Hz,1H),7.82–7.59(m,2H),5.06(s,2H),4.95(d,J＝4.4Hz,1H),4.87(s,1H),4.61(s,1H),3.66(s,1H),3.46(s,1H),3.14(d,J＝39.7Hz,4H).¹³C NMR(101MHz,DMSO-d₆)δ183.87,159.21,148.94,146.66,140.87,126.55,125.85,85.70,78.67,77.41,72.34,69.64,60.73。C₁₃H₁₇ClN₄O₅PtS(M+H⁺) Theoretical value 572.9040, test value 573.0031.

Example 5

Glycosylated platinum (II) Compounds antitumor Activity test

The experimental process comprises the following steps: the MTT colorimetric method has high sensitivity, and can be widely applied to activity detection of bioactive factors, large-scale antitumor drug screening, cytotoxicity tests, tumor radiosensitivity determination, tumor cell drug resistance determination and the like. The MTT method is used for determining the growth inhibition capacity of the glycosylated platinum compound on non-small cell lung cancer A549 and H460, breast cancer MCF-7, cervical cancer HeLa cells and the like. The method comprises the following basic steps: collecting tumor cells in logarithmic growth phase, counting trypsinized cells, inoculating the cells into a 96-well plate at a cell density of 3000-5000 cells per well, and culturing at 37 ℃ with 5% CO₂Under the constant condition, the mixture is incubated overnight, after the medicines with different concentration gradients are added, the mixture acts for 24 hours or 48 hours, MTT is added, and the culture is continued for 4 hours. Then, the original culture broth was discarded, and 100. mu.L of DMSO was added to each well. Succinate dehydrogenase in mitochondria of living cells can reduce exogenous MTT to water-insoluble blue-purple formazan and leave it in the cells, while dead cells cannot produce blue-purple formazan. Dimethyl sulfoxide was able to lyse formazan produced by cells and its light absorption (OD value) was determined with a microplate reader at a wavelength of 570 nm. A larger OD value indicates a relatively stronger cell activity and less drug toxicity. After the test is finished, the following equation can be obtained: cell survival (%) ═ (OD)_treated–OD_blank)/(OD_control–OD_blank) X 100%. Semitained IC for sensitivity of cells to drugs₅₀And expressing the value. Data processing and growth curve plotting were performed using Graphpad software.

The experimental results are as follows:

as shown in Table 2, the representative compound acts on non-small cell lung cancer A549 cells and H460 cells, breast cancer MCF-7 cells and cervical cancer HeLa cells respectively, and after 48 hours, the representative compound has obvious inhibition effect on four cells, which indicates that the platinum compound has good anti-tumor activity on four cells and can be used as a candidate anti-cancer drug in the future.

Table 2 represents the half-maximal Inhibition (IC) of glycosylated platinum (II) compounds on 4 cell lines₅₀) Value of

The foregoing shows and describes the general principles and broad features of the present invention and advantages thereof. While the invention has been described with respect to the above embodiments, it will be understood by those skilled in the art that the invention is not limited to the above embodiments, which are described in the specification and illustrated only to illustrate the principles of the invention, but that various changes and modifications may be made without departing from the spirit and scope of the invention as defined by the appended claims. The scope of the invention is defined by the appended claims and equivalents thereof.