Glycoside compounds, methods for preparing the same and uses thereof

Abstract

The application provides a kind of glycoside compound and its preparation method and purposes, specifically, the application provides a kind of compound as described in formula I, R is hydroxyl or H;X is or n=0, 1 or 2;Y is selected from monosaccharide or disaccharide;Z is N or CH. The compound is good in water solubility, and precipitates in buffer solution, indicating that the compound can be used as a tracer, and has good effect.

Description

Technical Field

[0001] This invention relates to the field of medicinal chemistry, specifically to a class of glycoside compounds and their preparation methods and uses. Background Technology

[0002] Malignant tumors seriously endanger human health, and timely and thorough surgical removal of cancer cells is an important means of cancer treatment. During surgery, it is necessary to remove as many lymph nodes as possible to prevent cancer cell metastasis and reduce the chance of cancer recurrence. The sentinel lymph node (SLN) is the first lymph node to be removed in a metastatic tumor and is the earliest site of tumor-associated antigen and adaptive immune response. Using lymph node tracers during surgery to dissect lymph nodes in metastatic lesions can reduce tissue damage, shorten surgical time, and thoroughly remove lymph nodes. For non-metastatic lesions, targeted removal of the sentinel lymph node can avoid immunodeficiency caused by extensive resection. This has become one of the guiding principles of tumor resection surgery and is applicable to various cancers such as breast cancer, gastric cancer, and colorectal cancer. Currently, the sentinel lymph node tracers used clinically abroad are mainly radionuclide tracers (such as 99Tcm-sulfur colloid), which are expensive and require the use of radiation tracking equipment during surgery, severely limiting their clinical application. Currently, the only domestically available product of its kind is the nano-carbon tracer, but it accumulates and remains in the patient's body, cannot be metabolized, and poses a risk of embedding and blockage. The only tracer in clinical trials is mitoxantrone, which has high cytotoxicity and can easily cause local tissue necrosis. Therefore, developing tracers with good tracking effects and high safety has significant clinical and industrial value.

[0003] The glycoside-based lymphatic tracer of this invention possesses a blue-black appearance, good water solubility, forms nano-sized particles under physiological conditions, and exhibits excellent lymphatic targeting and tracing effects. Furthermore, it exhibits very low cytotoxicity, rapid in vivo metabolism, and a wide therapeutic window, making it a promising candidate drug for lymphatic tracer therapy. Summary of the Invention

[0004] The technical problem to be solved by the present invention is to overcome the problem of the limited variety of existing tracers. The present invention provides a class of glycoside compounds, their preparation methods and uses. Specifically, it provides a compound as shown in Formula I, which is highly water-soluble and precipitates in buffer solution, indicating that the compound in this application can be used as a tracer and has good effect.

[0005] This invention provides a compound as shown in Formula I or a pharmaceutically acceptable salt thereof:

[0006]

[0007] R is a hydroxyl group or H;

[0008] X is n = 0, 1, or 2;

[0009] Y is selected from monosaccharide or disaccharide;

[0010] Z can be either N or CH.

[0011] In one embodiment, the compounds represented by Formula I may be defined as follows, and the remaining groups may be defined as described in any of the above embodiments:

[0012] n = 0.

[0013] In one embodiment, the compounds represented by Formula I may be defined as follows, and the remaining groups may be defined as described in any of the above embodiments:

[0014] When X is In this case, the b-terminus is preferably connected to Y, and correspondingly, the a-terminus is connected to the methylene group.

[0015] In one embodiment, the compounds represented by Formula I may be defined as follows, and the remaining groups may be defined as described in any of the above embodiments:

[0016] When X is In this case, the d-terminus is preferably connected to Y, and correspondingly, the c-terminus is connected to the methylene group.

[0017] In one embodiment, the compounds represented by Formula I may be defined as follows, and the remaining groups may be defined as described in any of the above embodiments:

[0018] When Y is a monosaccharide, the monosaccharide has an open-ring structure or a cyclic structure. The cyclic structure is preferably a five-membered ring or a six-membered ring, and more preferably a six-membered ring.

[0019] In one embodiment, the compounds represented by Formula I may be defined as follows, and the remaining groups may be defined as described in any of the above embodiments:

[0020] When Y is a monosaccharide, the monosaccharide is preferably xylose, mannosyl, lythose, galactosyl, sorbitol, arabinose, glucosyl, fructose, rhamnose, fucose, sialic acid, ribosyl, deoxyribosyl, alloose, arabinose, 2-aminoglucosyl, 2-aminogalactosyl, 2-amino-galactosyl, 2-amino-mannosyl, 2-aminoethyl-rhamnose, 2-aminoethyl-fucosyl, 4,6-O-ethylidene-D-glucosyl, 3-O-methyl-glucosyl, 4 6-O-benzyl-D-glucosyl, N-acetylglucosyl or N-acetylglucosyl, more preferably D-ribosyl, D-xylose, D-galactosyl, D-lythoseyl, D-glucosyl, D-arabinoseyl, D-2-glucosyl, D-2-glucosyl, D-mannosyl, L-fucosyl, D-3-O-methyl-glucosyl, 2-amino-D-galactosyl, 2-amino-D-mannosyl, 2-aminoethyl-D-rhamnosyl or 2-aminoethyl-D-fucosyl, for example For example Optimal Selection

[0021] In one embodiment, the compounds represented by Formula I may be defined as follows, and the remaining groups may be defined as described in any of the above embodiments:

[0022] When Y is a disaccharide, the monosaccharide in the disaccharide has an open-ring structure or a cyclic structure. The cyclic structure is preferably a five-membered ring or a six-membered ring, and more preferably a six-membered ring.

[0023] In one embodiment, the compounds represented by Formula I may be defined as follows, and the remaining groups may be defined as described in any of the above embodiments:

[0024] When Y is a disaccharide, the disaccharides in the disaccharide are, independently, xylose, mannose, lysose, galactose, sorbose, arabinose, glucose, fructose, rhamnose, fucose, sialic acid, ribose, deoxyribose, allose, arabinose, 2-glucosamine, 2-galactosamine, 2-amino-galactose, 2-amino-mannose, 2-aminoethyl-rhamnose, 2-aminoethyl-fucose, 4,6-O-ethylidene-D-glucose, 3-O-methyl-glucose, 4,6-O-benzylidene-D-glucose. - One or two of glucose, N-acetylglucosamine, and N-acetylglucosamine, more preferably one or two of D-ribose, D-xylose, D-galactose, D-lythose, D-glucose, D-arabinose, D-2-glucosamine, D-2-galactosamine, D-mannose, L-fucose, D-3-O-methyl-glucose, 2-amino-D-galactose, 2-amino-D-mannose, 2-aminoethyl-D-rhamnose, and 2-aminoethyl-D-fucose, wherein the disaccharide is preferably...

[0025] In one embodiment, the compounds represented by Formula I may be defined as follows, and the remaining groups may be defined as described in any of the above embodiments:

[0026] X is Preferred The b-terminus is preferably connected to Y, and correspondingly, the a-terminus is connected to methylene.

[0027] In one embodiment, the compounds represented by Formula I may be defined as follows, and the remaining groups may be defined as described in any of the above embodiments:

[0028] R is H.

[0029] In one embodiment, the compounds represented by Formula I may be defined as follows, and the remaining groups may be defined as described in any of the above embodiments:

[0030] Y represents a monosaccharide.

[0031] In one embodiment, the compounds represented by Formula I may be defined as follows, and the remaining groups may be defined as described in any of the above embodiments:

[0032] Z stands for CH.

[0033] In one embodiment, the compound represented by Formula I is preferably any of the following embodiments:

[0034] Option 1:

[0035] R is H;

[0036] n = 0;

[0037] Y is a monosaccharide group, and the monosaccharide has a cyclic structure, preferably a six-membered ring.

[0038] Option 2:

[0039] R is H;

[0040] X is The b-terminus is preferably connected to Y, and correspondingly, the a-terminus is connected to the methylene group;

[0041] n = 0;

[0042] Y is

[0043] Option 3:

[0044] R is H;

[0045] X is The b-terminus is preferably connected to Y, and correspondingly, the a-terminus is connected to the methylene group; n = 0;

[0046] Y is

[0047] In a preferred embodiment, the compound represented by Formula I is any of the following compounds:

[0048]

[0049]

[0050]

[0051] The present invention also provides a method for preparing the compound shown in Formula I, comprising the following steps: in a solvent, in the presence of a catalyst and a ligand, the compound shown in Formula II and the compound shown in Formula III undergo a click reaction as shown below to obtain the compound shown in Formula I.

[0052]

[0053] The conditions and steps of the click reaction described herein are conventional conditions and steps for this type of reaction in the art, and the present invention particularly prefers the following:

[0054] In the click reaction, the solvent is preferably an amide solvent, more preferably N,N-dimethylformamide.

[0055] In the click reaction, the catalyst is preferably a monovalent copper catalyst (e.g., Cu2SO4), which can be obtained by reducing divalent copper to a monovalent copper catalyst, for example, reducing CuSO4 with sodium ascorbate to obtain the monovalent copper catalyst Cu2SO4.

[0056] In the click reaction, the ligand is preferably 2-(4-((di((1-(tert-butyl-1H-1,2,3-triazol-4-yl)methyl)amino)methyl)-1H-1,2,3-triazol-1-yl)acetic acid.

[0057] The method for preparing the compound shown in Formula I may further include the following steps: in a solvent, the compound shown in Formula IV undergoes a substitution reaction with propargylamine as shown below to obtain the compound shown in Formula II;

[0058]

[0059] M is a halogen.

[0060] The conditions and steps of the substitution reaction are conventional conditions and steps for this type of reaction in the art, and the present invention particularly prefers the following:

[0061] In the substitution reaction described above, M is preferably fluorine, chlorine, bromine or iodine, more preferably fluorine.

[0062] In the substitution reaction, the solvent is preferably an ether solvent, and more preferably tetrahydrofuran.

[0063] In the substitution reaction, the reaction temperature is preferably 60-100°C, more preferably 80°C.

[0064] The present invention also provides a compound as shown in Formula II:

[0065]

[0066] R represents H or a hydroxyl group, and Z represents N or CH.

[0067] The present invention also provides a pharmaceutical composition comprising substance X and pharmaceutical excipients, wherein substance X is a compound as shown in Formula I or a pharmaceutically acceptable salt thereof. The amount of substance X is preferably a therapeutically effective amount.

[0068] The present invention also provides the use of the above-mentioned substance X or the above-mentioned pharmaceutical composition in the preparation of lymphatic tracers.

[0069] The present invention also provides the use of the above-mentioned substance X or the above-mentioned pharmaceutical composition as a lymphatic tracer.

[0070] Unless otherwise stated, the following terms appearing in this specification and claims have the following meanings:

[0071] The pharmaceutical excipients described herein are those widely used in the pharmaceutical manufacturing industry. Excipients primarily serve to provide a safe, stable, and functional pharmaceutical composition, and may also provide methods to facilitate the dissolution of the active ingredient at a desired rate after administration to a subject, or to promote the effective absorption of the active ingredient after administration of the composition to a subject. The pharmaceutical excipients may be inert fillers, or provide a function such as stabilizing the overall pH of the composition or preventing the degradation of the active ingredient. The pharmaceutical excipients may include one or more of the following: buffers, chelating agents, preservatives, solubilizers, stabilizers, excipients and surfactants, colorants, flavoring agents, and sweeteners.

[0072] In this invention, the term "pharmaceutically acceptable salt" refers to a salt formed with inorganic acids such as phosphoric acid, sulfuric acid, and hydrochloric acid, or organic acids such as acetic acid, tartaric acid, citric acid, and malic acid, or acidic amino acids such as aspartic acid and glutamic acid, or a salt formed with the above acids as an ester or amide and then with an inorganic base, such as sodium, potassium, calcium, aluminum, and ammonium salts.

[0073] The salts of the glycoside compounds described in this invention include, for example, maleate, hydrochloride, sulfate, acetate, trifluoroacetate, citrate, malate, tartrate, etc.

[0074] The term "halogen" refers to fluorine, chlorine, bromine, or iodine.

[0075] The term glycosyl refers to a sugar in which the hydroxyl group has been replaced.

[0076] The term open-ring structure refers to a branched or straight-chain structure that does not contain structures such as glucose that can form cyclic tautomers.

[0077] The term monosaccharide refers to a sugar whose molecular structure contains 3 to 6 carbon atoms; the term disaccharide refers to a disaccharide formed by the ether bond between the two hydroxyl groups of two monosaccharides.

[0078] The term five-membered ring refers to a ring formed by five atoms through chemical bonds, while the term six-membered ring refers to a ring formed by six atoms through compound bonds.

[0079] In this application The keys include both flat keys and straight keys.

[0080] Without violating common sense in the field, the above-mentioned preferred conditions can be combined arbitrarily to obtain various preferred embodiments of the present invention.

[0081] The reagents and raw materials used in this invention are all commercially available.

[0082] The positive and progressive effects of this invention are as follows: the glycoside compounds in this application have good water solubility and precipitate in buffer solutions, indicating that the compounds in this application can be used as tracers and have good effects. Attached Figure Description

[0083] Figure 1 The solubility of compound S4 in the solvent: Figure 1 A represents the solubility of compound S4 in water. Figure 1 B and Figure 1 C represents the solubility of compound S4 in the buffer solution.

[0084] Figure 2 The solubility of compound S21 in solvent: Figure 2 A represents the solubility of compound S21 in water. Figure 2 B represents the solubility of compound S21 in the buffer solution.

[0085] Figure 3 The solubility of compound T1 in the solvent: Figure 3 A represents the solubility of compound T1 in water. Figure 3 B and Figure 3 C represents the solubility of compound T1 in the buffer solution.

[0086] Figure 4 The solubility of compound T2 in the solvent: Figure 4 A represents the solubility of compound T2 in water. Figure 4 B and Figure 4 C represents the solubility of compound T2 in the buffer solution.

[0087] Figure 5 This represents the solubility of compound T3 in water.

[0088] Figure 6 This represents the solubility of compound T4 in water.

[0089] Figure 7 This represents the solubility of compound T5 in water.

[0090] Figure 8 The solubility of compound T6 in solvent: Figure 8 A represents the solubility of compound T6 in water. Figure 8 B and Figure 8 C represents the solubility of compound T6 in the buffer solution.

[0091] Figure 9 The solubility of compound T7 in solvent: Figure 9 A represents the solubility of compound T7 in water. Figure 9 B and Figure 9C represents the solubility of compound T7 in the buffer solution.

[0092] Figure 10 The solubility of compound T8 in solvent: Figure 10 A represents the solubility of compound T8 in water. Figure 10 B and Figure 10 C represents the solubility of compound T8 in the buffer solution.

[0093] Figure 11 This shows the solubility of compound T9 in water.

[0094] Figure 12 This is the particle size distribution diagram of SZ0126.

[0095] Figure 13 This is a plot showing the average particle size of midoanthraquinone.

[0096] Figure 14 The average particle size diagram of SZ0126 is shown.

[0097] Figure 15 Tests were conducted on the cytotoxicity of SZ0126, its hydrochloride, and mitoxantrone against HEK293 and CHO cells.

[0098] Figure 16 The image shows the staining results of SZ0126 and midoanthraquinone on mouse lymph.

[0099] Figure 17 This is a blood concentration-time curve for SZ0126. Detailed Implementation

[0100] The present invention is further illustrated below by way of embodiments, but the invention is not limited to the scope of the embodiments described herein. Experimental methods in the following embodiments that do not specify specific conditions were performed according to conventional methods and conditions, or as selected according to the product instructions.

[0101] For the following examples, standard operating and purification methods known to those skilled in the art can be used. Unless otherwise specified, the raw materials are generally available from commercially available sources such as Aldrich Chemicals Co. and AcrosOrganics. Commercially available solvents and reagents are generally used without further purification. Unless otherwise stated, all temperatures are expressed in °C (degrees Celsius), and room temperature or ambient temperature refers to 20–25 °C. The structures of the compounds were determined by nuclear magnetic resonance (NMR) spectroscopy and / or mass spectrometry (MS).

[0102] The shift (δ) of the proton NMR spectrum is given in parts per million (ppm). The proton NMR spectra are obtained using Bruker (AV) spectra. -The measurements were performed using a 500MHz nuclear magnetic resonance spectrometer, with deuterated dimethyl sulfoxide (DMSO-d6) as the solvent and tetramethylsilane (TMS) as the internal standard.

[0103] High-resolution mass spectrometry was performed using an Agilent 6230 series TOF LC-MS instrument.

[0104] HPLC: Agilent 1260 analytical high-performance liquid chromatography system (Agilent Technologies). Analytical high-performance liquid chromatography conditions: C18 column (5 μm, 4.6 x 250 mm), UV detection band 214, elution conditions: 0-90% acetonitrile (containing 0.1% v / v TFA) gradient wash for 30 min.

[0105] Chromatography columns typically use 200-300 mesh silica gel as a support.

[0106] In the above discussion and the following embodiments, the following abbreviations have the following meanings. If an abbreviation is not defined, it has the generally accepted meaning.

[0107] DMF is N,N-dimethylformamide;

[0108] PE is petroleum ether;

[0109] EA is ethyl acetate;

[0110] TFA stands for trifluoroacetic acid;

[0111] DMC is 2-chloro-1,3-dimethylimidazoline chloride.

[0112] BTTAA is 2-(4-((di((1-(tert-butyl-1H-1,2,3-triazol-4-yl)methyl)amino)methyl)-1H-1,2,3-triazol-1-yl)acetic acid

[0113] CuSO4·5H2O is copper sulfate pentahydrate.

[0114] Preparation of intermediates

[0115] Preparation of 1,6,9-propargylamine-difluorobenzisoquinoline-5,10-dione (1)

[0116]

[0117] Weigh 200 mg (0.8 mmol) of 6,9-difluorobenzisoquinoline-5,10-dione into a 50 mL round-bottom flask, then add 20 mL of acetonitrile and mix well. Add propargylamine (525 mL, 8.2 mmol) while stirring, and heat to 80°C for 16 h. During the reaction, the solution gradually changes from red to purple and finally to blue. After evaporating the solvent, perform column chromatography with PE:EA = 4:1 elution. Evaporation of the solvent yields a blue solid (108 mg, 42.0%). High-resolution mass spectrometry (ESI) N 19 H 14 N3O2, [M+H] + The theoretical value is 316.1081, and the measured value is 316.1085. 1 H NMR(500MHz,DMSO-d6)δ10.70(d,J=32.8Hz,2H),9.43(s,1H),9.00(t,J=4.8Hz,1H) ,8.06(d,J=5.1Hz,1H),7.71–7.54(m,2H),4.40(s,4H),3.32(dt,J=3.8,1.9Hz,2H).

[0118] Example 1

[0119]

[0120] In the first step, commercially available D-xylose (37.5 mg, 0.25 mmol) and sodium azide (162.5 mg, 2.5 mmol) were dissolved in 0.65 mL of double-distilled water. Then, triethylamine (0.35 mL) and DMC (127 mg, 0.75 mmol) were added separately, and the mixture was stirred in an ice bath for 1 hour. This solution was then used directly in the second step of the reaction.

[0121] In the second step, 4 μL of CuSO4·5H2O (100 mM), 12 μL of BTTAA (200 mM), and 5.6 μL of sodium ascorbate (1 M) were added sequentially. During this process, the solution changed from light blue to dark blue and then to colorless. Then, 200 μL (0.05 mmol) of 1-azido-D-xylose prepared in the first step was added and mixed thoroughly. Next, the intermediate 6,9-propyne-difluorobenzisoquinoline-5,10-dione (4 mg, 0.0125 mmol) was added, followed by 200 μL of DMF. The mixture was incubated in a 37°C water bath for 1 hour. The reaction was monitored by HPLC. HPLC: C18 column (5 μm, 4.6 x 250 mm), UV detection band 214 nm, elution conditions: 2-90% acetonitrile (containing 0.1% v / v TFA) gradient wash for 30 minutes. The crude product was then prepared by liquid chromatography (HPLC). 18C Flash column), freeze-dried to give a deep blue solid S1 (7.5 mg, 81.2%); high-resolution mass spectrometry (ESI+) C 29 H 32 N9O 10 [M+H] + The theoretical value is 666.2272, and the measured value is 666.1970; [M+Na] + The theoretical value is 688.2092, and the measured value is 688.1787. 1 H NMR (500MHz, DMSO-d6) δ11.21(s,1H),11.12(s,1H),9.41(d,J=7.3Hz,1H),8.96(d,J=5.2Hz,1H),8.30(s,2H),8.05(t,J=5.7Hz,1H),7 .73–7.63(m,2H),5.50(dd,J=9.3,0.9Hz,2H),4.81(d,J=6.9Hz,5H),3.88–3.74(m,4H),3.48(td,J=9.5,5.2Hz,2H),3.39–3.30(m,4H). 13 C NMR (126MHz, DMSO-d6) δ152.53,148.67,146.80,146.29,144.56,144.45,139.51,127.62, 126.23,125.49,122.71,118.76,109.67,88.62,77.52,72.42,69.56,68.78,38.21,38.17.

[0122] Example 2

[0123]

[0124] In Example 1, D-xylose was replaced with D-galactose. The remaining raw materials, reagents, and preparation methods were the same as in Example 1, yielding S2 (7.1 mg, 77%). High-resolution mass spectrometry (ESI+) C 31 H 36 N9O 12 [M+H] + The theoretical value is 726.2483, and the measured value is 726.2212. 1H NMR (500MHz, DMSO-d6) δ11.25(s,1H),11.16(s,1H),9.44(d,J=3.6Hz,1H),8.97(dd,J=5.2,2.5Hz,1H),8.25(d,J=3.9Hz,2H),8.06(d,J=5 .1Hz,1H),7.93–7.67(m,2H),5.51(dd,J=9.2,2.7Hz,2H),4.88–4.77(m,4H),4.64–3.88(m,2H),3.72(t,J=6.1Hz,2H),3.58–3.41(m,6H).

[0125] 13 C NMR (126MHz, DMSO-d6) δ180.48,152.83,148.88,146.84,146.33,144.61,144.50,139.45,126. 37,125.63,122.41,118.68,109.63,108.91,88.61,78.93,74.14,69.80,68.93,60.89,38.16.

[0126] Example 3

[0127]

[0128] In Example 1, D-xylose was replaced with D-ribose. The remaining raw materials, reagents, and preparation methods were the same as in Example 1, yielding S3 (6.4 mg, 76.2%). High-resolution mass spectrometry (ESI+) C 29 H 32 N9O 10 The theoretical value of [M+H]+ is 666.2272, and the measured value is 666.2159. 1 H NMR (500MHz, DMSO-d6) δ11.22(s,1H),11.12(s,1H),9.41(s,1H),8.97(d,J=5.2Hz,1H),8.32(d,J=41.2Hz,2H),8.05(d,J=5.2H z,1H),7.80–7.51(m,2H),5.65(d,J=9.6Hz,2H),4.82(d,J=6.9Hz,4H),4.04–3.94(m,4H),3.77–3.66(m,3H),3.62–3.49(m,1H).

[0129] 13C NMR (126MHz, DMSO-d6) δ180.36,179.36,152.62,148.73,146.81,146.30,144.55,144.44,139. 50,125.51,122.81,118.73,109.67,108.94,85.39,71.53,70.86,69.60,66.87,65.61,38.21.

[0130] Example 4

[0131]

[0132] In Example 1, D-xylose was replaced with D-lythose. The remaining raw materials, reagents, and preparation methods were the same as in Example 1, yielding S4 (5.5 mg, 65.5%). High-resolution mass spectrometry (ESI+) C 29 H 32 N9O 10 [M+H] + The theoretical value is 666.2272, and the measured value is 666.2260. 1 H NMR (500MHz, DMSO-d6) δ11.23(s,1H),11.13(s,1H),9.41(d,J=2.9Hz,1H),8. 99–8.94(m,1H),8.28(s,2H),8.08–8.02(m,1H),7.75–7.68(m,2H),5.69(d,J =9.2Hz,2H),4.83(d,J=7.2Hz,5H),4.27–4.21(m,2H),4.11(s,1H),4.00–3.9 3(m,2H),3.91(d,J=7.0Hz,1H),3.91(s,1H),3.71–3.61(m,5H),3.39(s,1H). 13 C NMR (126MHz, DMSO-d6) δ146.82,146.30,144.51,144.41,139.50,127.64,126. 28,125.54,122.65,118.76,108.92,85.89,71.42,69.60,67.73,67.15,38.21.

[0133] Example 5

[0134]

[0135] In Example 1, D-xylose was replaced with D-glucose. The remaining raw materials, reagents, and preparation methods were the same as in Example 1, yielding S5 (5.5 mg, 59.7%). High-resolution mass spectrometry (ESI+) C 31 H 36 N9O 12 [M+H] + The theoretical value is 726.2483, and the measured value is 726.2352. 1 H NMR (500MHz, DMSO-d6) δ11.20(s,1H),11.10(s,1H),9.39(d,J=7.5Hz,1H),8.95(d,J=5.2Hz,1H),8.32(d,J=5.1Hz,2H),8.04(dd,J=8. 7,5.2Hz,1H),7.76–7.65(m,2H),5.57(d,J=9.3Hz,2H),4.81(d,J=7.1Hz,3H),3.86–3.66(m,5H),3.55–3.36(m,7H),3.31–3.17(m,3H). 13 C NMR (126MHz, DMSO-d6) δ180.12,179.11,152.26,148.52,146.82,146.29,144.42,144.31,139.58,127. 66,126.27,125.53,122.89,118.85,109.60,108.84,87.99,80.44,77.43,72.53,70.04,61.19,38.15.

[0136] Example 6

[0137]

[0138] In Example 1, D-xylose was replaced with D-arabinose. The remaining raw materials, reagents, and preparation methods were the same as in Example 1, yielding S6 (6.7 mg, 81.7%). High-resolution mass spectrometry (ESI+) C 29 H 32 N9O 10 [M+H] + The theoretical value is 666.2272, and the measured value is 666.2260. 1H NMR (500MHz, DMSO-d6) δ11.23(s,1H),11.13(s,1H),9.42(d,J=9.1Hz,1H),8.96(d,J=5.2Hz,1H),8.24(s,2H),8.05(d,J=5.1Hz,1H),7.76–7.67(m ,2H),5.44(d,J=9.1Hz,2H),4.83(d,J=7.8Hz,5H),4.04(td,J=9.2,1.6H z,2H),3.87–3.80(m,1H),3.80–3.74(m,5H),3.57(dd,J=9.3,3.3Hz,2H). 13 C NMR(126MHz,DMSO-d6)δ180.32,152.58,148.72,146.82,146.30,144.60,144.50,139.50,1 27.64,126.28,125.53,122.38,118.77,109.65,108.91,88.89,73.68,69.79,68.82,38.19.

[0139] Example 7

[0140]

[0141] In Example 1, D-xylose was replaced with D-2-glucosamine. All other required raw materials, reagents, and preparation methods were the same as in Example 1. S7 (6.5 mg, 70.1%) was obtained using high-resolution mass spectrometry (ESI+). 31 H 38 N 11 O 10 [M+H] + The theoretical value is 724.2803, and the measured value is 724.2748. 1 H NMR (500MHz, DMSO-d6) δ11.31–11.11(m,2H),9.45(s,1H),9.00(s,1H),8.46(s,1H),8.06(d,J=5.0Hz,1H),7.79(q,J=6.5,4.7Hz, 2H),6.41(d,J=5.5Hz,1H),5.99(d,J=9.6Hz,1H),4.88(dt,J=12.2,5.9Hz,4H),4.46–3.56(m,6H),3.40–3.17(m,4H),2.64(s,1H). 13C NMR(126MHz,DMSO-d6)δ180.71,179.71,158.79,158.53,149.00,146.81,146.30,145.16,139.30,125.90, 123.95,84.20,82.09,80.62,76.98,73.43,70.71,70.51,69.77,60.65,54.47,53.10,44.94,38.19,31.65.

[0142] Example 8

[0143]

[0144] In Example 1, D-xylose was replaced with D-2-galactosamine. The remaining raw materials, reagents, and preparation methods were the same as in Example 1. S8 (7.2 mg, 78.3%) was obtained using high-resolution mass spectrometry (ESI+). 31 H 38 N 11 O 10 [M+H] + The theoretical value is 724.2803, and the measured value is 724.2758. 1 H NMR(500MHz,DMSO-d6)δ11.22(d,J=40.9Hz,1H),9.44(s,1H),8.99(d,J=5.3Hz,1H) ,8.41(s,1H),8.32(s,1H),8.05(dd,J=5.7,2.4Hz,1H),7.79(h,J=5.6,5.1Hz,2H),6 .43(d,J=5.6Hz,1H),5.93(d,J=9.6Hz,1H),4.87(dt,J=11.9,5.9Hz,4H),4.58(dt,J =11.1,2.4Hz,1H),3.54(td,J=14.0,12.5,6.2Hz,3H),3.41(dd,J=11.0,6.2Hz,1H). 13C NMR(126MHz,DMSO-d6)δ180.73,179.74,153.25,149.03,146.79,146.29, 139.29,127.50,126.03,125.63,123.91,118.55,109.02,84.63,82.48,7 9.09,75.73,70.76,67.67,67.44,67.37,60.61,60.51,51.57,49.61,44. 94,38.17,38.05,31.74,29.48,29.27,29.15,29.03,27.02,22.55,14.41.

[0145] Example 9

[0146]

[0147] In Example 1, D-xylose was replaced with D-mannose. The remaining raw materials, reagents, and preparation methods were the same as in Example 1, yielding S9 (6.5 mg, 70.1%). High-resolution mass spectrometry (ESI+) C 31 H 36 N9O 12 [M+H] + The theoretical value is 726.2483, and the measured value is 726.2473.

[0148] Example 10

[0149]

[0150] In Example 1, D-xylose was replaced with L-fucose. The remaining raw materials, reagents, and preparation methods were the same as in Example 1, yielding S10 (5.4 mg, 61.4%). High-resolution mass spectrometry (ESI+) C 31 H 36 N9O 10 [M+H] + The theoretical value is 694.2585, and the measured value is 694.2382.

[0151] Example 11

[0152]

[0153] In Example 1, D-xylose was replaced with D-3-oxo-methyl-glucose. The remaining raw materials, reagents, and preparation methods were the same as in Example 1, yielding S11 (7.3 mg, 74.0%). High-resolution mass spectrometry (ESI+) C 33 H 40 N9O 12The theoretical value of [M+H]+ is 754.2796, and the measured value is 754.2599.

[0154] Example 12

[0155]

[0156] In Example 1, D-xylose was replaced with D-merinobisaccharide. The remaining raw materials, reagents, and preparation methods were the same as in Example 1, yielding S12 (7.3 mg, 74.0%). High-resolution mass spectrometry (ESI+) C 43 H 56 N9O 22 [M+H] + The theoretical value is 1050.3540, and the measured value is 1050.3223.

[0157] Example 13

[0158]

[0159] In the first step, commercially available 2-amino-D-glucose (44.8 mg, 0.25 mmol), imidazole sulfonyl azide (80.5 mg, 0.375 mmol), potassium carbonate (69.2 mg, 0.5 mmol), and CuSO4·5H2O (6.3 mg, 0.025 mmol) were weighed and added to a 10 mL reaction flask. Then, 0.8 mL of methanol and 0.2 mL of dd H2O were added, and the mixture was stirred overnight at room temperature. This solution was then used directly in the second step of the reaction.

[0160] In the second step, 4 μL of CuSO4·5H2O (100 mM), 12 μL of BTTAA (200 mM), and 5.6 μL of sodium ascorbate (1 M) were added sequentially. During this process, the solution changed from light blue to dark blue and then to colorless. Then, 200 μL of 2-azido-D-glucose (0.05 mmol) prepared in the first step was added and mixed thoroughly. After that, the intermediate 6,9-propynylamine-difluorobenzisisoquinoline-5,10-dione (4 mg, 0.0125 mmol) was added, and 200 μL of DMF was added. The mixture was then incubated in a constant temperature water bath at 37 degrees Celsius for 1 h. The reaction was monitored by HPLC. HPLC: C18 column (5 μm, 4.6 x 250 mm), UV detection band was 214 nm, elution conditions: 2-90% acetonitrile (containing 0.1% v / v TFA) gradient wash for 30 min. The crude product was prepared by reverse-phase C18 chromatography and freeze-dried to obtain a deep blue solid S13 (7.0 mg, 76.1%); high-resolution mass spectrometry (ESI+) C13... 31 H 36 N9O 12 The theoretical value of [M+H]+ is 726.2483, and the measured value is 726.2265.

[0161] Example 14

[0162]

[0163] The 2-amino-D-glucose in Example 13 was replaced with 2-amino-D-galactose. The remaining raw materials, reagents, and preparation methods were the same as in Example 13, yielding S14 (6.7 mg, 72.8%). High-resolution mass spectrometry (ESI+) C 31 H 36 N9O 12 [M+H] + The theoretical value is 726.2483, and the measured value is 726.2249.

[0164] Example 15

[0165]

[0166] The 2-amino-D-glucose in Example 13 was replaced with 2-amino-D-mannose. The remaining raw materials, reagents, and preparation methods were the same as in Example 13, yielding S15 (5.7 mg, 62.0%). High-resolution mass spectrometry (ESI+) C 31 H 36 N9O 12 [M+H] + The theoretical value is 726.2483, and the measured value is 726.2246.

[0167] Example 16

[0168]

[0169] The 2-amino-D-glucose in Example 13 was replaced with 2-aminoethyl-D-rhamnose. The remaining raw materials, reagents, and preparation methods were the same as in Example 13, yielding S16 (9.9 mg, 65.7%). High-resolution mass spectrometry (ESI+) C 31 H 36 N9O 12 [M+H] + The theoretical value is 782.3109, and the measured value is 782.2905.

[0170] Example 17

[0171]

[0172] The 2-amino-D-glucose in Example 13 was replaced with 2-aminoethyl-D-fucose. The remaining raw materials, reagents, and preparation methods were the same as in Example 13, yielding S17 (8.8 mg, 58.4%). High-resolution mass spectrometry (ESI+) C 31 H 36 N9O12 [M+H] + The theoretical value is 782.3109, and the measured value is 782.2874.

[0173] Preparation of II 1,4-bis(propargylamino)anthraquinone (2)

[0174]

[0175] Weigh 200 mg (0.8 mmol) of 1,4-difluoroanthraquinone into a 50 mL round-bottom flask, then add 4 mL of tetrahydrofuran and mix well. Add 2 mL of propargylamine while stirring, and heat to 80°C for 16 h. During the reaction, the solution gradually changes from red to purple and finally to blue. After evaporating the solvent, perform column chromatography with PE:EA = 4:1 elution. Evaporation yields a blue solid, 1,4-di(propargylamino)anthraquinone II (108 mg, 42.0%). High-resolution mass spectrometry (ESI+) N 20 H 15 N₂O₂, [M+H] + The theoretical value is 315.1128, and the measured value is 315.1085. 1 H NMR (500MHz, DMSO) δ10.50 (t, J=5.9Hz, 2H), 8.24 (dd, J=5.8, 3.3Hz, 2H), 7.83 (dd, J=5.9,3.3Hz,2H),7.54(s,2H),4.36(dd,J=5.9,2.3Hz,4H),3.29(t,J=2.3Hz,2H).

[0176] Example 18

[0177]

[0178] In the first step, commercially available D-xylose (37.5 mg, 0.25 mmol) and sodium azide (162.5 mg, 2.5 mmol) were dissolved in 0.65 mL of dd water. Then, triethylamine (0.35 mL) and DMC (127 mg, 0.75 mmol) were added separately, and the mixture was stirred in an ice bath for 1 hour. This solution was then used directly in the second step reaction.

[0179] In the second step, 4 μL of CuSO4·5H2O (100 mM), 12 μL of BTTAA (200 mM), and 5.6 μL of sodium ascorbate (1 M) were added sequentially. During this process, the solution changed from light blue to dark blue and then to colorless. Then, 200 μL of 1-azido-D-xylose (0.05 mmol) prepared in the first step was added and mixed thoroughly. After that, the intermediate 1,4-bis(propargylamino)anthraquinone (4 mg, 0.0125 mmol) was added, and 200 μL of DMF was added. The mixture was then incubated in a constant temperature water bath at 37 degrees Celsius for 1 h. The reaction was monitored by HPLC. HPLC: C18 column (5 μm, 4.6 x 250 mm), UV detection band was 214 nm, elution conditions: 2-90% acetonitrile (containing 0.1% v / v TFA) gradient wash for 30 min. The crude product was prepared by reverse-phase C18 chromatography, and freeze-dried to obtain a deep blue solid S18 (7.5 mg, 81.2%); high-resolution mass spectrometry (ESI+) C18... 30 H 33 N8O 10 [M+H] + The theoretical value is 665.2314, and the measured value is 665.2350.

[0180] Example 19

[0181]

[0182] In Example 18, D-xylose was replaced with D-ribose, and the remaining raw materials, reagents, and preparation methods were the same as in Example 18, yielding S19 (3 ​​mg, 48%); high-resolution mass spectrometry (ESI+) C 30 H 33 N8O 10 [M+H] + The theoretical value is 665.2314, and the measured value is 665.2359.

[0183] Example 20

[0184]

[0185] The D-xylose in Example 18 was replaced with D-lythose. The remaining raw materials, reagents, and preparation methods were the same as in Example 18, yielding S20 (5.5 mg, 65.5%). High-resolution mass spectrometry (ESI+) C 30 H 33 N8O 10 [M+H] + The theoretical value is 665.2314, and the measured value is 666.2260.

[0186] Example 21

[0187]

[0188] The D-xylose in Example 18 was replaced with D-glucose. The remaining raw materials, reagents, and preparation methods were the same as in Example 18, yielding S21 (5.5 mg, 59.7%). High-resolution mass spectrometry (ESI+) C 32 H 37 N8O 12 [M+H] + The theoretical value is 725.2525, and the measured value is 725.2552. 1 H NMR (500MHz, DMSO) δ11.23(d,J=5.8Hz,1H),11.14(d,J=5.8Hz,1H),9.42(s,1H),8.97(d,J=5.1Hz,1 H),8.31(s,2H),8.04(d,J=5.1Hz,1H),7.77(q,J=10.0Hz,2H),5.54(d,J=9.2Hz,2H),5.41(dd,J=6.0 ,1.8Hz,2H),5.29(d,J=5.0Hz,2H),5.16(d,J=5.5Hz,2H),4.83(t,J=6.2Hz,4H),4.65(t,J=5.1Hz,2H ),3.83–3.72(m,2H),3.72–3.62(m,3H),3.43(d,J=7.3Hz,4H),3.40–3.36(m,3H),3.25–3.17(m,2H).

[0189] Example 22

[0190]

[0191] The D-xylose in Example 18 was replaced with D-mannose. The remaining raw materials, reagents, and preparation methods were the same as in Example 18, yielding S22 (6.5 mg, 70.1%). High-resolution mass spectrometry (ESI+) C 32 H 37 N8O 12 [M+H] + The theoretical value is 725.2525, and the measured value is 725.2493.

[0192] Example 23: Analysis of the physicochemical properties of the compound

[0193] The physicochemical properties of the above compounds are shown in Table 1:

[0194] The steps for preparing the stock solution are as follows: Add the test compound in the table below to water to form a stock solution with a concentration of 20 mg / mL, and observe the precipitation.

[0195] The steps for dilution using buffer solutions are shown in Table 1 below:

[0196] After observing the precipitation, take 20 μL of each of the above-mentioned compounds with good water solubility (20 mg / mL) and add them to 180 μL of phosphate buffer solution (pH = 7.4, 50 mM). Observe the precipitation, then centrifuge at high speed for 15 minutes and observe the precipitation again.

[0197] For a compound to produce good in vivo imaging, it needs to be highly water-soluble in aqueous solution but precipitate in buffer solution to achieve good imaging results.

[0198] (1) Properties of crystallization in water-soluble, neutral pH buffer solutions.

[0199] Crystals precipitate in water-soluble solutions and in blood pH (i.e., buffer solutions with a pH of 7.4).

[0200] Table 1

[0201]

[0202]

[0203]

[0204] (2) Particle size analysis:

[0205] The particle size distribution of SZ0126 is shown below. Figure 12 :

[0206] It is known that the crystal particles precipitated from SZ0126 in the solvent have nanoparticle properties and passive lymphatic targeting function.

[0207] For the average particle size distribution of mitogen anthraquinone, please refer to [link / reference]. Figure 13 .

[0208] For the average particle size diagram of SZ0126, please refer to [link / details]. Figure 14 .

[0209] Example 24 Compound Toxicity Analysis

[0210] Cytotoxicity test

[0211] Cellular-level toxicity tests were performed on the compounds of this invention. Compound concentrations were diluted according to the 2006 CLSI (Clinical Laboratory Standards Institute) dilution method, and their toxicity was assessed. Results are as follows: Figure 15 As shown.

[0212] Assay method: HEK293 cells and CHO cells were seeded into 96-well plates. The test compound was dissolved in a certain volume of DMSO to prepare a 10 mM solution, which was used as the initial concentration. This solution was then diluted 3-fold with cell culture medium. 10 μL of each concentration of the compound was added to the cells and incubated at 37°C with 5% CO2 for 48 h. MTT was then added, and the cells were incubated at 37°C for 4 h. Finally, 100 μL of triplet solution was added, and the cells were incubated overnight. OD values ​​were read at 570 nm and 690 nm using a microplate reader.

[0213] Depend on Figure 15 It can be seen that SZ0126 and its hydrochloride have almost no effect on the activity of HEK293 cells and CHO cells, while mitoanthraquinone significantly reduces the activity of HEK293 cells and CHO cells.

[0214] Mouse toxicity test:

[0215] Local toxicity:

[0216] The SZ0126 group, the mitoxantrone group, and the saline group each contained 3 SD rats.

[0217] SZ0126 was injected subcutaneously into the hind legs of each animal (2 mg / hind leg, 1 mg / hind leg, 0.05 mL). The animals were administered SZ0126 once a week for four consecutive weeks, followed by a 14-day observation period after drug withdrawal.

[0218] Observe the injection site daily for any abnormal reactions such as edema or ulceration, and record the weight of each rat and the amount of food consumed by each group of rats.

[0219] After the observation period ended, the rats were euthanized by cervical dislocation, and pathological sections (HE staining) were prepared from the drug administration site.

[0220] Experimental results:

[0221] Group SZ0126 was in normal condition, with no deaths or wound ulceration.

[0222] In the mitoxantrone group, some rats died and their wounds ulcerated in week 1, and all rats died in week 2. The specific experimental results of the rat body weight and mortality are shown in Table 2 below:

[0223] Table 2

[0224]

[0225] Example 25: Lymphatic Tracing Effect of Compound

[0226] Mouse lymph node staining experiment

[0227] Three Bal / bc mice were included in each of the following groups: SZ0126 group (3 mg / mL, 1 mg / mL), mitoxantrone group (1 mg / mL), and saline group. Each group received an injection of 100 μL of SZ0126, mitoxantrone, or saline via the paw, respectively. One mouse in each group was euthanized by cervical dislocation at 15 min, 30 min, and 60 min after administration. Lymph nodes were dissected and their staining was observed. The staining results are shown below. Figure 16 .

[0228] Example 26 Pharmacokinetic Experiment

[0229] (1) Reagents: Glutinous glycoside reference standard was purchased from Shanghai Yuanye Biotechnology Co., Ltd.; acetonitrile, formic acid and acetonitrile were purchased from Merck, Germany; other reagents were of analytical grade.

[0230] (2) Chromatographic column

[0231] Column: BDS Hypersil C 18 Chromatographic column (2.1 mm × 50 mm, 2.4 μm, Thermo); mobile phase: acetonitrile (A) containing 0.2% formic acid : water containing 0.2% formic acid (B), flow rate: 0.35 mL / min, gradient elution: 0.0 min–0.5 min, 20% A; 0.5 min–0.8 min, 20% A–90% A; 0.8 min–3.3 min, 90% A–20% A; 3.3 min–4.5 min, 20% A. Column temperature: 40 °C, injection volume: 4 μL.

[0232] (3) Administration

[0233] Rats were subcutaneously injected with 5 mg / kg SZ0126, and the drug concentration in the rat blood was tested at different time points. The blood drug concentration-time curve is shown in the figure below. Figure 17 As shown.

[0234] (4) Sample pretreatment methods

[0235] Take 100 μL of plasma sample, add 300 μL of acetonitrile solution containing internal standard (30 ng / ml acetonitrile) as protein precipitant, vortex for 5 min, centrifuge at 13000 rpm for 10 min, transfer 300 μL of supernatant to a new EP tube, concentrate under vacuum to dryness, add 75 μL of 50% acetonitrile, vortex for 5 min, centrifuge at 13000 rpm for 10 min, aspirate the supernatant to the sample vial, and test the drug concentration in plasma.

[0236] Combination Figure 17 The blood drug concentration-time curve was calculated using the blood drug concentrations at the first five points (0.5h, 1h, 2h, 4h, 8h), and the specific parameters are shown in Table 3 below:

[0237] Table 3

[0238] AUC(0-t) ng / mL*h 2663.92 4433.9 4146.3 3748.04 949.824 AUC(0-∞) ng / mL*h 2673.82 4607.557 4248.405 3843.261 1028.563 AUMC(0-t) h*h*ng / mL 4182.86 8348.2 7787.4 6772.82 2260.43 AUMC(0-∞) h*h*ng / mL 4275.689 10134.731 8815.077 7741.832 3073.432 MRT(0-t) h 1.57 1.883 1.878 1.777 0.179 MRT(0-∞) h 1.599 2.2 2.075 1.958 0.317 <![CDATA[t 1 / 2 from]]> h 0.954 1.585 1.431 1.323 0.329 Tmax h 0.5 0.5 0.5 0.5 0 CLz / F L / h / kg 1.87 1.085 1.177 1.377 0.429 Cmax ng / mL 1270 1810 1790 1623.333 306.159

[0239] Note:

[0240] AUC: Area under the curve during drug administration; AUMC: Area under the curve of first moment; MRT: Mean residence time; t 1 / 2 Half-life; Tmax: Time to peak concentration; CL: Clearance; F: Bioavailability; Cmax: Peak concentration.

[0241] The parameters mentioned above, when denoted by z, are statistical moment parameters; when denoted by 'z', they are room-compartment model parameters.

[0242] The above description is merely a specific embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any changes or substitutions that can be conceived by those skilled in the art within the scope of the technology disclosed in the present invention without creative effort should be included within the scope of protection of the present invention.

Claims

1. A compound as shown in Formula I or a pharmaceutically acceptable salt thereof: I R is H; X is n = 0, 1, or 2; Y is a monosaccharide group; Z is either N or CH. The monosaccharide is , , , , , , , , , , , , , , , , , , , or .

2. The compound of formula I as claimed in claim 1, or a pharmaceutically acceptable salt thereof, characterized in that: n=0； And / or, the b-end of X is connected to Y, and correspondingly, the a-end is connected to the methylene group; And / or, the monosaccharide is or .

3. The compound of formula I as claimed in claim 1, or a pharmaceutically acceptable salt thereof, characterized in that: The b-terminus of X is connected to Y, and correspondingly, the a-terminus is connected to the methylene group; And / or, Z is CH.

4. The compound of formula I as claimed in claim 1, or a pharmaceutically acceptable salt thereof, characterized in that, The compound represented by Formula I is any one of the following compounds: , , , , , , , , , , , , , , , , , , , or .

5. A method for preparing a compound of formula I as described in any one of claims 1 to 4, characterized in that, It includes the following steps: in a solvent, in the presence of a catalyst and a ligand, the compound shown in Formula II undergoes a click reaction with the compound shown in Formula III as shown below to obtain the compound shown in Formula I. 。 6. A pharmaceutical composition comprising substance X and a pharmaceutical excipient, wherein substance X is a compound of formula I as described in any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof.

7. The pharmaceutical composition of claim 6, wherein the amount of substance X is a therapeutically effective amount.

8. The use of a substance X or a pharmaceutical composition as described in claim 6 or 7 in the preparation of a lymphatic tracer, wherein the substance X is a compound of formula I as described in any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof.

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