CN113943330A - Sugar-containing structure compound, preparation method, pharmaceutical composition and application thereof - Google Patents
Sugar-containing structure compound, preparation method, pharmaceutical composition and application thereof Download PDFInfo
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Abstract
The invention relates to a compound containing a sugar structure shown as the following general formula I, a preparation method, a pharmaceutical composition and application thereof. Experiments prove that the compound has certain activity on inhibiting the interaction of PD-1/PD-L1 protein, and partial compound can effectively promote the secretion of IFN-gamma and can become a potential drug for treating tumors by inhibiting PD-L1. In addition, compared with the similar compound without the sugar structure, the compound with the sugar structure has obviously reduced cytotoxicity, thus having better safety.
Description
Technical Field
The invention belongs to the field of medicinal chemistry, and particularly relates to a PD-1/PD-L1 protein interaction small molecule inhibitor and pharmaceutically acceptable salts thereof, a method for preparing the PD-L1 small molecule inhibitor, a pharmaceutical composition containing the PD-L1 small molecule inhibitor derivative or pharmaceutically acceptable salts thereof, and application of the PD-L1 small molecule inhibitor derivative and pharmaceutically acceptable salts thereof in preparation of medicines for treating tumors or autoimmune related diseases.
Background
The exploration of immunotherapy has been over a hundred years, and recently, successful cases that the immunosuppressive targets such as CTLA-4, TIM-3, LAG-3 and PD-1/PD-L1 can be effectively treated by clinically inhibiting the immunosuppressive targets provide certain key support for searching immunotherapy of tumors or other diseases.
The main protein target in the immunosuppressive targets is the interaction of PD-1 and PD-L1. The PD-1(CD279) protein expressed on the surface of the T cell can transduce inhibitory signals by combining with the receptor PD-L1(CD274) or PD-L2(CD273), thereby inhibiting the activation of the T cell and reducing the immunity of the antiviral or tumor cells in the body. Due to the immunosuppressive effect of PD-1/PD-L1, T cell activation is inhibited on the surface of many tumor cells by expressing PD-L1 protein, thereby avoiding immune surveillance in the body. Clinical researches in recent years show that the interaction of PD-1/PD-L1 protein is blocked, so that the T cell fatigue phenomenon can be effectively reversed, tumor cells can be efficiently killed, and the success of immunotherapy is improved. PD-1 inhibitory antibodies that have been currently approved by the FDA for marketing are primarily nivolumab (Opdivo, Bristol Myers Squibb) and pembrolizumab (Keytruda, Merck), while PD-L1 inhibitory antibodies are primarily atezolizumab (Teentriq, Genentech/Roche), durvalumab (Imfnzi, AstraZeneca) and avelumab (Bavencio, EMD Serono, Inc.). The PD-1/PD-L1 inhibiting antibodies have already achieved good curative effect on tumor by being combined with anti-tumor drugs clinically at present. However, the antibody drug has poor penetrability in tumor infiltrates, particularly solid tumor tissues, and is high in preparation and cost, and the small-molecule inhibitor has the advantages of low cost, high stability, strong penetrability and low immunogenicity, so that the development of the PD-L1 small-molecule inhibitor has a great development space.
According to the invention, a series of sugar-containing small molecules with novel structures are designed and synthesized, so that the compound has good PD-L1 inhibitory activity and tumor immunotherapy effect in PD-1/PD-L1 blocking experiments, in-vitro immunity enhancement evaluation, in-vivo anti-tumor evaluation and other experiments, and has low cytotoxicity and good development prospect.
Disclosure of Invention
The invention aims to provide a novel compound containing a sugar structure, which has PD-L1 inhibitory activity.
The invention also aims to provide a preparation method of the compound containing the sugar structure.
Another object of the present invention is to provide a pharmaceutical composition, nutraceutical composition or food composition comprising the sugar structure-containing compound.
The invention also aims to provide application of the sugar-containing structural compound or the pharmaceutical composition containing the sugar-containing structural compound as a small molecule inhibitor of PD-L1 and application of the sugar-containing structural compound or the pharmaceutical composition containing the sugar-containing structural compound in treating tumor diseases and immune diseases.
In one aspect, the present invention provides a compound of the following general formula I:
wherein:
x is selected from N or C-R5，
R5Selected from H, halogen, nitro, cyano, selected from C1-C6Amino substituted by 1 or 2 substituents in a linear or branched alkyl radical, C1-C5Straight or branched alkyl, C1-C5Straight or branched alkoxy or C1-C5A linear or branched alkylthio group; preferably, R5Selected from H, halogen, nitro, C1-C5Straight or branched alkyl or C1-C5A linear or branched alkoxy group; more preferably, R5Selected from H, methyl, ethyl, propyl, fluorine, chlorine, nitro, methoxy and ethoxy
A group or propoxy group;
R1and R2Are respectively and independently selected from H, halogen, nitro,Cyano, selected from C1-C5Amino substituted by 1 or 2 substituents in a linear or branched alkyl radical, C1-C5Straight or branched alkyl, C1-C5Straight or branched alkoxy, C1-C5Straight or branched alkylthio or-CH2NH-(CH2)m-Y-Z, and R1And R2At least one is selected from-CH2NH-(CH2)m-Y-Z; or, R1And R2Each independently selected from H, halogen, C1-C5Straight or branched alkyl, C1-C5Straight or branched alkoxy or-CH2NH-(CH2)m-Y-Z, and R1And R2At least one is selected from-CH2NH-(CH2)m-Y-Z; or, R1And R2Each independently selected from H, methyl, ethyl, methoxy, ethoxy, fluorine, chlorine or-CH2NH-(CH2)m-Y-Z, and R1And R2At least one is selected from-CH2NH-(CH2)m-Y-Z wherein
m is an integer between 1 and 5, preferably 1,2,3 or 4;
y is absent or is-S-, -O-, -NR6-、-C(O)NH-、-NHC(O)-、-(CH2)q-or a triazole, which is a nitrogen-containing compound,
wherein:
q is an integer between 1 and 2;
R6selected from H, C1-C5A linear or branched alkyl group;
preferably, Y is-O-, -NH-, -C (O) NH-, -NHC (O) -,
z is selected from a modified or unmodified monosaccharide or disaccharide residue;
R3and R4Each independently selected from H, halogen, nitro, cyano, from C1-C6Amino substituted by 1 or 2 substituents in a linear or branched alkyl radical, C1-C5Straight or branched alkyl, C1-C5Straight or branched alkoxy or C1-C5A linear or branched alkylthio group; preferably, R3And R4Each independently selected from H, halogen, nitro, C1-C5Straight or branched alkyl or C1-C5A linear or branched alkoxy group; more preferably, R3And R4Each independently selected from H, methyl, ethyl, propyl, fluoro, chloro, nitro, methoxy, ethoxy or propoxy;
in one embodiment, in formula I,
x is selected from N or C-R5，
Wherein, when X is N,
R1is-CH2NH–(CH2)m-Y-Z wherein
m is an integer between 1 and 5, preferably 1,2,3 or 4;
y is-O-, -NH-, -C (O) NH-, -NHC (O) -, or triazole, specifically, Y is-O-, -NH-, -C (O) NH-, -NHC (O) -, or,
Z is selected from a modified or unmodified monosaccharide or disaccharide residue;
R2、R3and R4Each independently selected from H, halogen, nitro, cyano, C1-C5 straight or branched chain alkyl or C1-C5 straight or branched chain alkoxy, preferably, R2、R3And R4Each independently selected from H, methyl, ethyl, propyl, fluoro, chloro, nitro, methoxy, ethoxy or propoxy,
wherein, when X is C-R5When the temperature of the water is higher than the set temperature,
R1、R2and R4Each independently selected from H, halogen, nitro, cyano, C1-C5 linear or branched alkyl, C1-C5 linear or branched alkoxy, or-CH2NH–(CH2)m-Y-Z, and R1、R2And R4At least one of them is selected from-CH2NH–(CH2)m-Y-Z; or, R1、R2And R4Each independently selected from H, halogen, C1-C5Straight or branched alkyl, C1-C5Straight or branched alkoxy or-CH2NH–(CH2)m-Y-Z, and R1、R2And R4At least one is selected from-CH2NH–(CH2)m-Y-Z; or, R1、R2And R4Each independently selected from H, methyl, ethyl, methoxy, ethoxy, fluorine, chlorine or-CH2NH–(CH2)m-Y-Z, and R1、R2And R4At least one is selected from-CH2NH–(CH2)m-Y-Z wherein
m is an integer between 1 and 5, preferably 1,2,3 or 4;
y is-O-, -NH-, -C (O) NH-, -NHC (O) -, or triazole; specifically, Y is-O-, -NH-, -C (O) NH-, -NHC (O) -,
z is selected from a modified or unmodified monosaccharide or disaccharide residue;
R3and R5Each independently selected from H, halogen, nitro, cyano, C1-C5 straight or branched chain alkyl or C1-C5 straight or branched chain alkoxy; preferably, R3And R5Each independently selected from H, methyl, ethyl, propyl, fluoro, chloro, nitro, methoxy, ethoxy or propoxy.
In a preferred embodiment, the mono-or disaccharide residue is selected from the following:
in one embodiment, in formula I,
x is selected from N or C-R5，
R1is-CH2NH–(CH2)m-Y-Z, wherein m, Y and Z are as previously defined;
R2and R4Each independently selected from H, halogen, C1-C5 straight or branched chain alkyl or C1-C5 straight or branched chain alkoxy; preferably, R2And R4Each independently selected from H, methyl, ethyl, propyl, methoxy, ethoxy or propoxy; more preferably, R2And R4Each independently selected from H, methyl or methoxy;
R3and R5Each independently selected from H, halogen, nitro, C1-C5 straight or branched chain alkyl or C1-C5 straight or branched chain alkoxy; preferably, R3And R5Each independently selected from H, halogen, nitro, methyl, ethyl, propyl, methoxy, ethoxy or propoxy, more preferably R3And R5Each independently selected from H, fluoro, chloro, nitro, methyl or methoxy.
In one embodiment, in formula I,
x is C-R5，
R1Selected from H or halogen;
R2is-CH2NH-(CH2)m-Y-Z, wherein m, Y and Z are as previously defined, R4Is H;
R3and R5Each independently selected from H, halogen, nitro, C1-C5 straight or branched chain alkyl or C1-C5 straight or branched chain alkoxy; preferably, R3And R5Each independently selected from H, halogen, nitro, methyl, ethyl, propyl, methoxy, ethoxy or propoxy, more preferably R3And R5Each independently selected from H, fluoro, chloro, nitro, methyl or methoxy.
In a preferred embodiment, the compound is selected from one of the following compounds:
in particular embodiments, the pharmaceutically acceptable salts include salts of the compounds of formula I with the following anions: cl-、Br-、OH-、CH3COO-、PO4 3-、HPO4 2-、H2PO4 -、HSO4 -、SO4 2-Maleate ion, citrate ion, malate ion, and tartrate ion.
In another aspect, the present invention provides a process for the preparation of the above compound, said process comprising the steps of:
subjecting the compound of the general formula d and an amine compound containing a monosaccharide or disaccharide residue to a reductive amination reaction to obtain a compound of the general formula I,
wherein, in the above general formula d, R1a、R2aAnd R4aOne is an aldehyde group, and when R is1aWhen it is an aldehyde group, R2aAnd R4aAre as defined for R above2And R4When R is defined as2aWhen it is an aldehyde group, R1aAnd R4aAre as defined for R above1And R4When R is defined as4aWhen it is an aldehyde group, R1aAnd R2aAre as defined for R above1And R2The definition of (1); in the compounds of the formula I, X, R1，R2，R3And R4Are as defined above, respectively.
In the above reaction, the solvent includes halogenated hydrocarbon solvents such as dichloromethane, 1, 2-dichloroethane, chloroform and the like, aromatic hydrocarbon solvents such as benzene, toluene and the like, aprotic solvents such as N, N-dimethylformamide, dimethylsulfoxide, hexamethylphosphoramide and the like, ether solvents such as tetrahydrofuran, diethyl ether, 1, 4-dioxane and the like, protic solvents such as water, methanol, ethanol and the like, or a mixture of these solvents.
In the above reaction, the base used includes organic bases such as triethylamine, pyridine, N-diisopropylethylamine, 4-dimethylaminopyridine, 1, 8-diazabicyclo [5.4.0] -7-undecene, 1,2,2,6, 6-pentamethylpiperidine and the like.
In the above reaction, the reducing agent used includes sodium borohydride, sodium cyanoborohydride, sodium triethoxyborohydride and sodium triacetoxyborohydride.
It will be appreciated that in the above routes, the precise order of the synthetic steps for introducing the various groups and moieties into the molecule may vary. The skilled person will be able to ensure that the groups or moieties introduced in one stage of the process will not be affected by subsequent transformations and reactions and will select the order of the synthetic steps accordingly.
In another aspect, the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of one or more selected from the above-mentioned compounds or pharmaceutically acceptable salts thereof as an active ingredient, and optionally a pharmaceutically acceptable carrier, excipient, adjuvant and/or diluent.
In particular embodiments, the pharmaceutical composition may further comprise other pharmaceutically acceptable therapeutic agents.
In another aspect, the invention provides a use of the above compound or a pharmaceutically acceptable salt thereof, or the above pharmaceutical composition in the preparation of a PD-L1 inhibitor.
In another aspect, the invention provides a use of the above compound or a pharmaceutically acceptable salt thereof, or the above pharmaceutical composition in the preparation of a medicament for treating a tumor disease.
In another aspect, the present invention provides a use of the above compound or a pharmaceutically acceptable salt thereof, or the above pharmaceutical composition in the preparation of a medicament having an immunomodulatory effect.
In a specific embodiment, the immunomodulatory effect is achieved by inhibiting immunosuppression mediated by PD-L1.
Advantageous effects
The compound has certain activity on inhibiting the interaction of PD-1/PD-L1 protein, and partial compound can effectively promote the secretion of IFN-gamma and can become a potential drug for treating tumors by inhibiting PD-L1. Compared with the similar compound without the sugar structure, the compound with the sugar structure provided by the invention has the advantages that the cytotoxicity is obviously reduced, and the safety is better.
Drawings
FIG. 1 shows the test results of the partial compound of the present invention degrading PD-L1 protein.
FIG. 2 shows the results of the test of antitumor activity in a partial compound mouse of the present invention, wherein FIG. 2A shows the effect of the compound on the average tumor volume of tumor-bearing mice, and FIG. 2B shows the effect of the compound on the average body weight of tumor-bearing mice.
FIG. 3 shows the results of the effect of some compounds of the invention on Tumor Infiltrating Lymphocytes (TILs) in mice.
Detailed Description
The invention will be further illustrated in the following examples. These examples are intended to illustrate the invention only and do not limit the scope of protection of the invention in any way.
For the following examples, standard procedures and purification methods known to those skilled in the art may be used. Unless otherwise specified, starting materials are generally available from commercial sources, such as Aldrich Chemicals Co. and Acros Organics. Commercial solvents and reagents were generally used without further purification, anhydrous solvents were processed by standard methods, and other reagents were commercially available as analytical grade. Unless otherwise stated, all temperatures are expressed in degrees Celsius (C.) and room or ambient temperature means 20-25 ℃. The structure of the compounds is determined by nuclear magnetic resonance spectroscopy (NMR) and/or Mass Spectrometry (MS).
The nuclear magnetic resonance hydrogen spectral shift (δ) is given in parts per million (ppm). NMR spectra were measured on a Mercury-600MHz and Bruker (AV-400)400MHz NMR spectrometer, deuterated dimethyl sulfoxide (DMSO-d)6) Deuterated chloroform (CDCl)3) Deuterated methanol (MeOD-d)4) Deuterium oxide (D)2O) is used as solvent, and Tetramethylsilane (TMS) is used as internal standard.
High resolution mass spectrometry was determined using Agilent 6230 series TOF LC-MS and if the intensity of the chloride or bromide containing ion was described, the expected intensity ratio (including35Cl/37Ion of Cl about 3:1, comprising79Br/81Ion 1:1) of Br and only gives the intensity of ions of lower mass.
HPLC: agilent 1260 analytical high performance liquid chromatography system (Agilent). Analytical high performance liquid chromatography conditions: c18 column (5 μm,4.6X 250mm), UV detection band 214 and 280nm, elution conditions 0-90% acetonitrile (containing 0.1% v/v TFA) gradient for 30 min.
The chromatographic column generally uses 200-300 mesh silica gel as a carrier.
In the above discussion and in the examples below, the following abbreviations have the following meanings. An abbreviation has a generally accepted meaning if it is not defined.
TLC is thin layer chromatography
DIPEA is N, N-diisopropylethylamine;
NHS is N-hydroxysuccinimide;
EDCI is 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride;
DMF is N, N-dimethylformamide;
PE is petroleum ether;
DCM is dichloromethane;
EtOAc is ethyl acetate;
MsCl is methylsulfonyl chloride;
CsCO3is cesium carbonate;
Pd/C is palladium carbon;
TFA is trifluoroacetic acid;
Pd2(dba)3tris (dibenzylideneacetone) dipalladium;
X-PHOS is 2-dicyclohexyl phosphonium-2, 4, 6-triisopropyl biphenyl.
BTTAA is {4- [ (bis { [1- (2-methyl-2-propyl) -1H-1,2, 3-triazol-4-yl ] methyl } amino) methyl ] -1H-1,2, 3-triazol-1-yl } acetic acid.
Preparation of intermediates
Preparation example 1: preparation of 2, 6-dimethoxy-4- (2-methyl-3-phenylphenylmethoxy) benzaldehyde (1)
Methanesulfonyl chloride (2mL,25.8mmol) was slowly added dropwise to 40mL of a solution of 3-hydroxymethyl-2-methylbiphenyl 1a (4.3g,21.5mmol) and triethylamine (3.6mL,25.8mmol) in dichloromethane under ice-cooling, and after completion of the dropwise addition, the reaction was carried out at room temperature for 40 min. The reaction mixture was washed twice with a saturated aqueous sodium bicarbonate solution (10mL), distilled water (10mL) and a saturated aqueous sodium chloride solution (10mL) in this order, and then dried over anhydrous sodium sulfate as a solid. Filtering and concentrating to obtain a white solid crude product of the mesylate 1b of the 3-hydroxymethyl-2-methylbiphenyl, and directly carrying out the next reaction. Crude mesylate 1b of 3-hydroxymethyl-2-methylbiphenyl (2g,7.2mmol) and 2, 6-dimethoxy-4-hydroxybenzaldehyde (1.4g,7.7mmol) were dissolved in 30mL DMF, and anhydrous potassium carbonate solid (1.5g,10.8mmol) was added and stirred at room temperature overnight. The residue was concentrated to remove the solvent, slurried with petroleum ether (20mL), and filtered to give white 2, 6-dimethoxy-4- (2-methyl-3-phenylphenylmethoxy) benzaldehyde 1(1.95g, yield 75%). High resolution mass spectrometry (ESI +) C23H22O4Theoretical value of 363.1596, found value of 363.1568[ M + H ]]+。
Preparation example 2: preparation of 2, 6-dimethyl-4- (2-methyl-3-phenylphenylmethoxy) benzaldehyde (2)
The required raw materials, reagents and preparation method were the same as in preparation example 1 except that 2, 6-dimethoxy-4-hydroxybenzaldehyde in preparation example 1 was replaced with 2, 6-dimethyl-4-hydroxybenzaldehyde to obtain 2, 6-dimethyl-4- (2-methyl-3-phenylbenzyloxy) benzaldehyde 2.
1H NMR(500MHz,CDCl3):δ(ppm)＝10.47(s,1H),7.46–7.36(m,3H),7.37-7.21(m,5H),6.69(s,2H),5.11(s,2H),2.60(s,6H),2.22(s,3H).
13C NMR(500MHz,CDCl3):δ(ppm)＝191.68,162.06,144.59,143.09,141.82,134.58,134.38,130.41,129.39,128.13,128.02,126.95,126.22,125.69,115.58,77.23,69.07,29.73,22.71,21.17,16.20,14.15.
Preparation example 3: preparation of 3-fluoro-4- (2-methyl-3-phenylphenylmethoxy) benzaldehyde (3)
3-fluoro-4- (2-methyl-3-phenylbenzyloxy) benzaldehyde 3 was obtained in the same manner as in preparation example 1, except that 2, 6-dimethoxy-4-hydroxybenzaldehyde in preparation example 1 was replaced with 3-fluoro-4-hydroxybenzaldehyde.
1H NMR(500MHz,CDCl3):δ(ppm)＝9.80(d,J＝2.1Hz,1H),7.59-7.55(m,2H),7.38-7.33(m,3H),7.31-7.26(m,1H),7.26-7.22(m,2H),7.21-7.18(m,2H),7.13(t,J＝8.2Hz,1H),5.19(s,2H),2.19(s,3H).
13C NMR(500MHz,CDCl3):δ(ppm)＝189.92,153.86,152.30,151.87,143.16,141.69,134.30,133.84,130.58,130.35,129.39,128.10,127.85,127.00,125.74,115.98,114.35,70.45,16.21.
Preparation example 4: preparation of 3-chloro-4- (2-methyl-3-phenylphenylmethoxy) benzaldehyde (4)
3-chloro-4- (2-methyl-3-phenylbenzyloxy) benzaldehyde 4 was obtained in the same manner as in preparation example 1, except that 2, 6-dimethoxy-4-hydroxybenzaldehyde in preparation example 1 was replaced with 3-chloro-4-hydroxybenzaldehyde.
1H NMR(500MHz,CDCl3):δ(ppm)＝δ9.82(s,1H),7.90(d,J＝2.0Hz,1H),7.74(dd,J＝8.4,2.0Hz,1H),7.45(dd,J＝6.9,2.1Hz,1H),7.41-7.37(m,2H),7.34-7.30(m,1H),7.30-7.26(m,2H),7.25-7.21(m,2H),7.13(d,J＝8.4Hz,1H),5.23(s,2H),2.24(s,3H).
Preparation example 5: preparation of 3-nitro-4- (2-methyl-3-phenylphenylmethoxy) benzaldehyde (5)
3-Nitro-4- (2-methyl-3-phenylbenzyloxy) benzaldehyde 5 was obtained in the same manner as in preparation example 1, except that 2, 6-dimethoxy-4-hydroxybenzaldehyde in preparation example 1 was replaced with 3-nitro-4-hydroxybenzaldehyde.
1H NMR(500MHz,CDCl3):δ(ppm)＝9.92(s,1H),8.34(d,J＝2.0Hz,1H),8.06(dd,J＝8.7,2.1Hz,1H),7.44–7.38(m,3H),7.35–7.31(m,2H),7.29–7.25(m,2H),5.33(s,2H),2.24(s,3H).
13C NMR(500MHz,CDCl3):δ(ppm)＝188.77,156.25,143.23,141.52,140.27,134.60,134.06,132.94,130.69,129.40,129.19,128.19,127.49,127.46,127.06,125.85,114.97,70.97,16.25.
Preparation example 6: preparation of 3- (2-methyl-3-phenylphenylmethoxy) -4-methoxybenzaldehyde (6)
The required raw materials, reagents and preparation method were the same as in preparation example 1 except that 2, 6-dimethoxy-4-hydroxybenzaldehyde in preparation example 1 was replaced with 3-hydroxy-4-methoxybenzaldehyde, to give 3- (2-methyl-3-phenylbenzyloxy) -4-methoxybenzaldehyde 6.
1H NMR(500MHz,CDCl3):δ(ppm)＝9.83(s,1H),7.51(d,J＝1.9Hz,1H),7.47(dd,J＝8.2,1.9Hz,1H),7.43(dd,J＝7.2,1.9Hz,1H),7.39(m,2H),7.34–7.27(m,3H),7.23–7.19(m,2H),6.99(d,J＝8.2Hz,1H),5.17(s,2H),3.93(s,3H),2.24(s,3H).
13C NMR(500MHz,CDCl3):δ(ppm)＝191.02,155.25,148.93,142.91,141.96,134.71,134.27,130.21,130.01,129.46,128.09,127.89,127.21,126.86,125.63,111.19,110.79,70.02,56.20,16.18.
Preparation example 7: preparation of 3- (2-methyl-3-phenylphenylmethoxy) -4-chlorobenzaldehyde (7)
The required raw materials, reagents and preparation method were the same as in preparation example 1 except that 2, 6-dimethoxy-4-hydroxybenzaldehyde in preparation example 1 was replaced with 3-hydroxy-4-chlorobenzaldehyde, to give 3- (2-methyl-3-phenylbenzyloxy) -4-chlorobenzaldehyde 7.
1H NMR(500MHz,CDCl3):δ(ppm)＝9.93(s,1H),7.58-7.21(m,11H),5.21(s,2H),2.25(s,3H).
13C NMR(500MHz,CDCl3):δ(ppm)＝191.06,154.99,142.98,141.79,136.02,134.24,134.11,130.99,130.48,130.30,129.44,128.14,127.53,126.94,125.69,124.66,111.59,70.05,16.25.
Preparation example 8: preparation of 2-chloro-5- (2-methyl-3-phenylphenylmethoxy) benzaldehyde (8)
The required raw materials, reagents and preparation method were the same as in preparation example 1 except that 2, 6-dimethoxy-4-hydroxybenzaldehyde in preparation example 1 was replaced with 2-chloro-5-hydroxybenzaldehyde, to give 2-chloro-5- (2-methyl-3-phenylbenzyloxy) benzaldehyde 8.
1H NMR(500MHz,CDCl3):δ(ppm)＝10.33(s,1H),7.43(d,J＝3.1Hz,1H),7.33–7.10(m,9H),7.07(dd,J＝8.8,3.2Hz,1H),5.00(s,2H),2.11(s,3H).
13C NMR(500MHz,CDCl3):δ(ppm)＝189.81,157.90,143.12,141.79,134.40,132.96,131.62,130.47,130.09,129.40,128.14,128.02,126.96,125.71,123.56,112.80,69.66,16.22.
Preparation example 9: preparation of 2- (2-methyl-3-phenylphenylmethoxy) pyridine-5-carbaldehyde (9)
3-hydroxymethyl-2-methylbiphenyl 1a (2g,10.1mmol) and 2-chloropyridine-5-carbaldehyde 8a (1.4g,9.9mmol) were mixed with cesium carbonate (4.9g,15.1mmol), Pd2(dba)3(0.5g,0.5mmol) and X-PHOS (0.3g,0.6mmol) were mixed in toluene (30mL) and reacted at 85 ℃ for 8h under nitrogen. The residue was concentrated to remove the solvent, slurried with petroleum ether (15mL), and filtered to give off-white 2- (2-methyl-3-phenylphenylmethoxy) pyridine-5-carbaldehyde 9(2g, yield 65.3%).
1H NMR(500MHz,CDCl3):δ(ppm)＝9.97(s,1H),8.67(d,J＝2.3Hz,1H),8.10(dd,J＝8.7,2.3Hz,1H),7.47–7.40(m,3H),7.38–7.30(m,3H),7.29–7.25(m,2H),6.92(d,J＝8.6Hz,1H),5.55(s,2H),2.28(s,3H).
Preparation example 10: preparation of 2-methoxy-6- (2-methyl-3-phenylphenylmethoxy) pyridine-3-carbaldehyde (10)
2-methoxy-6-chloro-pyridine-3-carbaldehyde (10a)
After sec-butyllithium (5g,77mmol) was added to 60mL of tetrahydrofuran at-78 ℃ to obtain a uniform solution, 150mL of a tetrahydrofuran solution of 2-chloro-6-methoxypyridine (10g,70mmol) was slowly dropped. After stirring for 1h, DMF (7.5mL) was added dropwise slowly. After the reaction was carried out for another 1.5 hours, acetic acid (8.4mL) was slowly added dropwise to quench the reaction, the cooling bath was removed until the temperature of the reaction system naturally increased to 0 ℃ and then MTBE (100mL) and water (30mL) were added thereto to separate the layers. The organic layer was washed twice with saturated aqueous sodium hydrogencarbonate (30mL) and saturated brine (30mL) in this order, and then dried over anhydrous sodium sulfate as a solid. Filtering, concentrating to obtain crude product, adding 20mL n-hexane, pulping for 1h, and vacuum filtering to obtain light yellow 2-methoxy-6-chloro-pyridine-3-formaldehyde 10b solid (7.4g, 61.6%)
1H NMR(500MHz,CDCl3):δ(ppm)＝10.24(s,1H),8.00(d,J＝7.9Hz,1H),6.96(d,J＝7.7Hz,1H),4.03(s,3H).
2-methoxy-6- (2-methyl-3-phenylphenylmethoxy) pyridine-3-carbaldehyde (10)
The required starting materials, reagents and preparation were the same as in preparation example 9 except that 2-chloropyridine-5-carbaldehyde in preparation example 9 was replaced with 2-methoxy-6-chloro-pyridine-3-carbaldehyde, to give 2-methoxy-6- (2-methyl-3-phenylbenzyloxy) pyridine-3-carbaldehyde 10. High resolution mass spectrometry (ESI +) C23H22O4Theoretical value of 334.1443, found value of 334.1434[ M + H ]]+。
Preparation example 11: preparation of 2, 6-dimethoxy-4- (2-methyl-3-phenylphenylmethoxy) 2-propynyl-1-ylamine (11)
5.5mL of glacial acetic acid was slowly added to 20mL of a solution of 2, 6-dimethoxy-4- (2-methyl-3-phenylphenylmethoxy) benzaldehyde (1.0g) in DCM and propynylamine (0.306g) was added, after stirring well, sodium cyanoborohydride (0.352g) was added, after reaction at room temperature for 2h, the solvent was removed by distillation under the reduced pressure, and column chromatography was performed to give white 2, 6-dimethoxy-4- (2-methyl-3-phenylphenylmethoxy) 2-propynyl-1-ylamine 11(0.83g, yield 75%). High resolution mass spectrometry (ESI +) C26H27NO3Theoretical value of 402.2069, found value of 402.1913[ M + H ]]+。
Preparation example 12: preparation of 2-methoxy-6- (2-methyl-3-phenylphenylmethoxy) pyridin-3-propynyl-1-ylamine (12)
The required raw materials, reagents and preparation method were the same as in preparation example 11 except that 2, 6-dimethoxy-4- (2-methyl-3-phenylphenylmethoxy) benzaldehyde in preparation example 11 was replaced with 2-methoxy-6- (2-methyl-3-phenylphenylmethoxy) pyridine-3-carbaldehyde, to give 12. High resolution mass spectrometry (ESI)+):C24H24N2O2[M+H]+Theoretical value is 373.1916, found 373.1902.
Preparation example 13: preparation of 3- (2-methyl-3-phenylphenylmethoxy) -4-propynyl-1-ylamine (13)
Propyleneamine (0.25g) was slowly added to a DMF solution (20mL) of 3- (2-methyl-3-phenylphenylmethoxy) -4-methoxybenzaldehyde (1.0g), and 1.0mL of DIPEA was added, followed by reaction at 55 ℃ for 1 hour. The reaction was stopped, cooled to room temperature, and TFA was added to adjust pH 5. During the reaction, sodium cyanoborohydride (0.94g) was slowly added, and after reacting at room temperature for 2 hours, sodium cyanoborohydride (0.47g) was additionally added. After 4 hours of the reaction, the solvent was removed by distillation under the reduced pressure, and the resulting product was subjected to column chromatography to give 3- (2-methyl-3-phenylphenylmethoxy) -4-propynyl-1-ylamine 13(0.54g, yield 48.3%) as a white solid. High resolution mass spectrometry (ESI +) C25H25NO2Theoretical value of 372.1963, found value of 372.1977[ M + H ]]+。
Preparation example 14: preparation of 2-aminoethyl-beta-D-glucopyranoside (14)
1,2,3,4, 6-O-acetyl-D-glucopyranoside (14a)
D-glucose (5g,27.9mmol) was dissolved in 25ml pyridine and 25ml acetic anhydride was added slowly at 0 ℃ in an ice bath. Stirring at room temperature for 8h, evaporating under reduced pressure to remove solvent, dissolving the residual liquid in ethyl acetate, washing with 1N hydrochloric acid, pure water and saturated saline solution sequentially, drying the anhydrous sodium sulfate solid, filtering, and evaporating under reduced pressure to remove residual ethyl acetate. To give 1,2,3,4, 6-O-acetyl-D-glucopyranoside 14a (10.6g, yield 97%) as a transparent viscous oil,
2- (benzyloxycarbonyl) aminoethyl 2,3,4, 6-tetra-O-acetyl-. beta. -D-glucopyranoside (14b)
1,2,3,4, 6-O-acetyl-D-glucopyranoside 14a (4g,10.2mmol) and N- (benzyloxycarbonyl) ethanolamine (3.4g,16.7mmol) are dissolved in anhydrous acetonitrile (50mL), under nitrogen and in an ice bath a 48% strength boron trifluoride ether solution (5.7mL,22.2mmol) is slowly added dropwise. After stirring for 1h, the ice bath is removed, after reacting for 16h at room temperature, triethylamine is added for quenching. After the solvent was distilled off under reduced pressure, dichloromethane (50mL) was added and washed twice with saturated aqueous sodium bicarbonate (15mL), distilled water (15mL) and saturated brine (15mL) in this order, the anhydrous sodium sulfate solid was dried, filtered, and the residual dichloromethane was distilled off under reduced pressure and subjected to column chromatography (ethyl acetate/hexane 1:3 to 1:1, v/v) to give 2- (benzyloxycarbonyl) aminoethyl 2,3,4, 6-tetra-O-acetyl-. beta. -D-glucopyranoside 14b (2.6g, 48.5%).
2- (benzyloxycarbonyl) aminomethyl- β -D-glucopyranoside (14c)
2- (benzyloxycarbonyl) aminoethyl 2,3,4, 6-tetra-O-acetyl-. beta. -D-glucopyranoside 14b (0.5g,0.95mmol) was dissolved in a 4N methanol solution of sodium methoxide, and the mixture was reacted at room temperature for 3 hours while maintaining the pH at 10, followed by addition of a strong acid ion resin to adjust the pH at 7 to 8, filtration and evaporation of the solvent under reduced pressure to give 2- (benzyloxycarbonyl) aminomethyl-. beta. -D-mannoside 14c as a colorless oil (321.7mg, yield 94.7%).
2-aminoethyl-beta-D-glucopyranoside (14)
2- (benzyloxycarbonyl) aminomethyl- β -D-mannoside 14C (300mg,0.84mmol) was dissolved in methanol, 30mg of Pd-C (20%) was added, and H was introduced into the reaction mixture2(pressure 1.5Pa), stirred at room temperature for 3h, filtered through a sand-core funnel to give 2-aminoethyl- β -D-glucopyranoside 14(178.6mg, yield 95.2%).
2-aminoethyl-beta-D-glucopyranoside (14) high resolution mass spectrometry (ESI)+):C8H17NO6[M+H]+Theoretical value is 224.1134, found 224.1135.
Preparation example 15: preparation of 2-aminoethyl-beta-D-galactopyranoside (15)
The required starting materials, reagents and preparation were the same as in preparation example 14 except that glucose was replaced with galactose in preparation example 14 to give 15a-c and 15.
2-aminoethyl-beta-D-galactopyranoside (15) high-resolution Mass Spectrometry (ESI)+):C8H17NO6[M+H]+Theoretical value is 224.1134, found 224.1133.
Preparation example 16: preparation of 2-aminoethyl-alpha-D-mannopyranoside (16)
The required raw materials, reagents and preparation method were the same as in preparation example 14 except that glucose was replaced with mannose in preparation example 14, to give 16a-c and 16.
2-aminoethyl-alpha-D-mannopyranoside (16) high resolution Mass Spectrometry (ESI)+):C16H23NO8[M+Na]+Theoretical value is 380.1321, found 380.1316.
Preparation example 17: preparation of 2-aminoethyl-beta-D-xylopyranoside (17)
The required raw materials, reagents and preparation method were the same as in preparation example 14 except that glucose was replaced with xylose in preparation example 14, to give 17a-c and 17.
Preparation example 18: preparation of 2-aminoethyl-alpha-L-rhamnopyranoside (18)
The required starting materials, reagents and preparation were the same as in preparation example 14 except that glucose in preparation example 14 was replaced with rhamnose, to give 18a-c and 18.
1H NMR(400MHz,D2O):3.92-3.81(m,1H),3.73–3.54(m,3H),3.43(ddd,J＝10.4,6.1,4.0Hz,1H),3.34(t,J＝9.7Hz,1H),2.88–2.65(m,2H),1.19(d,J＝6.3Hz,3H).
Preparation example 19: preparation of 2-aminoethyl-beta-L-fucopyranoside (19)
The required raw materials, reagents and preparation method were the same as in preparation example 14 except that glucose in preparation example 14 was replaced with fucose, to give 19a-c and 19.
2-aminoethyl-beta-L-fucopyranoside (19) high resolution Mass Spectrometry (ESI)+):C8H17NO5[M+H]+Theoretical value is 208.1185, found 208.1306.
Preparation example 20: preparation of 2-aminoethyl-2-acetylamino-2-deoxy-beta-D-glucopyranoside (20)
1,3,4, 6-O-acetyl-2-acetamido-D-glucopyranoside (20a)
2-acetamido-D-glucopyranose (5g,22.6mmol) was dissolved in 25ml pyridine, ice-cooled at 0 ℃ and slowly added 25ml acetic anhydride. Stirring at room temperature for 8h, evaporating under reduced pressure to remove solvent, dissolving the residual liquid in ethyl acetate, washing with 1N hydrochloric acid, pure water and saturated saline solution sequentially, drying the anhydrous sodium sulfate solid, filtering, and evaporating under reduced pressure to remove residual ethyl acetate. 1,3,4, 6-O-acetyl-2-acetamido-D-glucopyranoside 20a (8.6g, yield 97.5%) was obtained as a pale yellow transparent viscous oil,
2- (benzyloxycarbonyl) aminoethyl-2-acetylamino-3, 4, 6-tetra-O-acetyl-beta-D-glucopyranoside (20b)
1,3,4, 6-O-acetyl-2-acetamido-D-glucopyranoside 20a (4g,10.3mmol) and N- (benzyloxycarbonyl) ethanolamine (3.5g,17.9mmol) were dissolved in anhydrous acetonitrile (50mL), and tin tetrachloride (5.7mL,22.2mmol) was slowly added dropwise under nitrogen protection and in an ice bath. Stirring for 30min, removing ice bath, reacting at room temperature for 30min, gradually heating to 75 deg.C, and stopping reaction after 24 hr. After the reaction temperature is naturally cooled to room temperature, triethylamine is added for quenching. After the solvent was distilled off under reduced pressure, dichloromethane (50mL) was added and washed twice with saturated aqueous sodium bicarbonate solution (15mL), distilled water (15mL) and saturated brine (15mL) in this order, the anhydrous sodium sulfate solid was dried, filtered, and the residual dichloromethane was distilled off under reduced pressure and subjected to column chromatography (ethyl acetate/hexane 1:1 to 3:1, v/v) to give 2- (benzyloxycarbonyl) aminoethyl-2-acetylamino-3, 4, 6-tetra-O-acetyl-. beta. -D-glucopyranoside 20b (3.2g, 59.2%).
2- (benzyloxycarbonyl) aminomethyl-2-acetylamino-beta-D-glucopyranoside (20c)
2- (benzyloxycarbonyl) aminoethyl-2-acetylamino-3, 4, 6-tetra-O-acetyl-. beta. -D-glucopyranoside 20b (0.6g,1.1mmol) was dissolved in a 4N methanol solution of sodium methoxide, and the reaction was carried out at room temperature for 3 hours while maintaining the pH at 10, followed by addition of a strong acid ion resin to adjust the pH at 7 to 8, filtration and evaporation of the solvent under reduced pressure to give 2- (benzyloxycarbonyl) aminomethyl-2-acetylamino-. beta. -D-glucopyranoside 20c (417.6mg, yield 95.3%).
2-aminoethyl-beta-D-glucopyranoside (20)
2- (benzyloxycarbonyl) aminomethyl-2-acetylamino-. beta. -D-glucopyranoside 20C (0.4mg,1.0mmol) was dissolved in methanol, 40mg of Pd-C (20%) was added thereto, and H was introduced into the reaction mixture2Stirring for 3h at room temperature under the pressure of 1.5Pa, and filtering with a sand core funnel to obtain 2-aminoethyl-beta-D-glucopyranoseGlycoside 20(252.8mg, yield 95.7%). High resolution mass spectrometry (ESI)+):C10H20N2O6Theoretical value of 265.1400, found value of 265.1404[ M + H ]]+。
Preparation example 21: preparation of N- (3-aminopropyl) -D-glucamide (21)
Gluconolactone (2g, 11.4mmol) was dissolved in 15mL of methanol, and N-tert-butoxycarbonyl-1, 3-propanediamine (2.37g,13.63mmol) was added. Refluxing for 4h, distilling off methanol under reduced pressure to obtain white solid 21a, washing with ethyl acetate, distilling off solvent under reduced pressure, dissolving 21a in 5mL of methanol, adding 4N hydrochloric acid, stirring at room temperature for 1h, distilling off solvent under reduced pressure to obtain N- (3-aminopropyl) -D-glucamide 21, and high resolution mass spectrometry (ESI)+)C9H20N2O6Theoretical value of 253.1400, found value of 253.1381[ M + H ]]+。
Preparation example 22: preparation of N- (3-aminopropyl) -D-lactase (22)
22a and 22 were obtained by using the same raw materials, reagents and production methods as in production example 21 except that gluconolactone in production example 21 was replaced with lactobionic acid.
High resolution mass spectrometry (ESI) of N- (3-aminopropyl) -D-lactase (22)+)C15H30N2O11Theoretical value of 415.1928, found value of 415.1901[ M + H ]]+。
Preparation example 23: preparation of 2-Aminon-propyl-beta-D-glucopyranoside (23)
The required starting materials, reagents and preparation were the same as in preparation example 14 except that N- (benzyloxycarbonyl) ethanolamine in preparation example 14 was replaced with N- (benzyloxycarbonyl) N-propanolamine, to give 23a-b and 23.
2-amino-n-propyl-beta-D-glucopyranoside (23) high resolution mass spectrum (ESI)+)C9H19NO6Theoretical value of 238.1290, found value of 238.1917[ M + H ]]+。
Preparation example 24: preparation of 2-amino-n-butyl-. beta. -D-glucopyranoside (24)
The required starting materials, reagents and preparation were the same as in preparation example 14 except that N- (benzyloxycarbonyl) ethanolamine in preparation example 14 was replaced with N- (benzyloxycarbonyl) N-butylaminoamine, to give 24a-b and 24.
2-amino n-butyl-beta-D-glucopyranoside (24) high resolution mass spectrum (ESI)+)C10H21NO6Theoretical value of 252.1447, found value of 252.2045[ M + H ]]+。
Preparation example 25: preparation of N- (3-aminoethyl) -D-lactase (25)
The required raw materials, reagents and preparation method were the same as in preparation example 21 except that N-t-butoxycarbonyl-1, 3-propanediamine in preparation example 21 was replaced with N-t-butoxycarbonyl-1, 3-ethanediamine, to give 25a and 25.
N- (3-aminoethyl) -D-lactase (25) high resolution Mass Spectrometry (ESI)+)C14H28N2O11Theoretical value of 401.1771, found value of 401.2239[ M + H ]]+。
Preparation example 26: preparation of N- (3-aminobutyl) -D-lactonamide (26)
The required raw materials, reagents and preparation method were the same as in preparation example 21 except that N-t-butoxycarbonyl-1, 3-propanediamine in preparation example 21 was replaced with N-t-butoxycarbonyl-1, 3-butanediamine, to give 26a and 26.
N- (3-Aminobutyl) -D-Lactamide (26) high resolution Mass Spectrometry (ESI)+)C16H32N2O11Theoretical value of 429.2084, found value of 429.2534[ M + H ]]+。
Preparation example 27: preparation of (2R,3R,4R,5S) -6- (2-Aminoethylamino) -1,2,3,4, 5-hexanpentaol (27)
D-glucose (5g,27.9mmol) was reacted with N-tert-butoxycarbonyl-1, 3-ethylenediamine (6.7g,41.9mmol) dissolved in methanol (50mL) at 60 ℃ for 1h, cooled to room temperature, adjusted to pH 6 with TFA, and sodium cyanoborohydride (3.5g,55.8mmol) was added. After reacting at room temperature for 3 hours, the solvent was distilled off under reduced pressure, and the crude product of 27a obtained was washed twice with methylene chloride, and 50mL of 2N HCl in methanol was added. After reacting for 1h, the reaction solution was distilled under reduced pressure to obtain (2R,3R,4R,5S) -6- (2-aminoethylamino) -1,2,3,4, 5-hexanpentol 27(4.9g, 79.6%) and high resolution mass spectrum (ESI)+)C8H20N2O5Theoretical value of 225.1540, found value of 225.1803[ M + H ]]+。
Preparation example 28: preparation of (2R,3R,4R,5S) -6- (2-aminopropylamino) -1,2,3,4, 5-hexanpentaol (28)
28a and 28 were obtained by the same procedures as in production example 27 except that N-t-butoxycarbonyl-1, 3-ethylenediamine in production example 27 was replaced with N-t-butoxycarbonyl-1, 3-propanediamine.
(2R,3R,4R,5S) -6- (2-aminopropylamino) -1,2,3,4, 5-hexanpentol (28) high resolution mass spectrum (ESI)+)C9H22N2O5Theoretical value of 239.1607, found value of 239.1610[ M + H ]]+。
Preparation example 29: preparation of 2- (. beta. -aminopropionylamino) -D-glucopyranose (29)
2- [ beta- (benzyloxycarbonyl) aminopropionylamino ] -D-glucopyranose (29a)
After N- (benzyloxycarbonyl) - β -alanine (2g,8.4mmol) and NHS (1.9g,16.8mmol), EDCI (3.2g,16.8mmol) and DIPEA (2.9mL,16.8mmol) were dissolved in dichloromethane (20mL) and reacted at room temperature for 4h, the reaction mixture was washed twice with distilled water (5mL) and saturated brine (5mL), respectively, and after drying the anhydrous sodium sulfate solid, the solvent was removed by suction filtration and distillation under reduced pressure. Yield 29a was dissolved in DMF (30mL) and glucosamine hydrochloride (2.0g,9.2mmol) and DTPEA (1.5mL,8.4mmol) were added. After 3h reaction at room temperature, the solvent was removed by autoclaving and the residue was washed twice with diethyl ether to give 2- [ β - (benzyloxycarbonyl) aminopropionylamino ] -D-glucopyranose 29a (2.5g, 78.1%).
2- (. beta. -aminopropionylamino) -D-glucopyranose (29)
2- [ beta- (benzyloxycarbonyl) aminopropionylamino]-D-glucopyranose 29a (1g,2.6mmol) was dissolved in methanol, 0.1g Pd-C (20%) was added, and H was passed through the reaction solution2Stirring at room temperature for 3h (pressure 1.5Pa), filtering with a sand core funnel to obtain 2- (2-aminopropionylamino) -D-glucopyranose 29(0.63g, 96.8%), and high resolution mass spectrometry (ESI)+)C9H18N2O6Theoretical value of 251.1243, found value of 251.1706[ M + H ]]+。
Preparation example 30: preparation of 2- (gamma-aminobutyrylamino) -D-glucopyranose (30)
The required starting materials, reagents and preparation were the same as in preparation 29 except that N- (benzyloxycarbonyl) - β -alanine in preparation 29 was replaced with N- (benzyloxycarbonyl) - γ -butyric acid, to give 30a and 30 a.
2- (gamma-aminobutyrylamino) -D-glucopyranose (30) high resolution mass spectrometry (ESI)+)C10H20N2O6Theoretical value of 265.1399, found value of 265.18695 [ M + H ]]+。
Preparation example 31: preparation of 1-beta-azido-4, 6-O-ethylene-D-glucose (31)
Dissolving 4, 6-O-ethylene-D-glucose (0.616g,3mmol) in a mixed system of 2mL of water and 8mL of acetonitrile under ice bath, adding 2mL of triethylamine after the sugar is fully dissolved, adding 2.6g of 2-azido-1, 3-dimethylimidazole hexafluorophosphate (9 mmol), stirring for 1h under ice bath, gradually returning to room temperature, stirring overnight, and storing the reaction system at 4 ℃ for later use.
Preparation example 32: preparation of 1-beta-azido-3-O-methylglucose (32)
Preparation 31 was repeated except that 3-O-methylglucose was used instead of 4, 6-O-ethylene-D-glucose in preparation 31, to give 32.
Preparation example 33: preparation of 1-beta-azido-4, 6-O-benzylidene-D-glucose (33)
33 was obtained by following the same procedures as in production example 31 except that 4, 6-O-ethylene-D-glucose was replaced with 4, 6-O-benzylidene-D-glucose in production example 31.
Preparation example 34: preparation of 1-beta-azido-D-glucose (34)
34 was obtained by following the same procedures as in preparation example 31 except that 4, 6-O-ethylene-D-glucose was replaced with D-glucose in preparation example 31.
Preparation example 35: preparation of 1-beta-azido-L-arabinose (35)
The required raw materials, reagents and preparation methods were the same as in preparation example 31 except that 4, 6-O-ethylene-D-glucose in preparation example 31 was replaced with L- (+) -arabinose, to give 35.
Preparation example 36: preparation of 1-beta-Azide-D- (+) -xylose (36)
The required raw materials, reagents and preparation methods were the same as in preparation example 31 except that 4, 6-O-ethylene-D-glucose in preparation example 31 was replaced with D- (+) -xylose, to give 36.
Preparation example 37: preparation of 1-beta-azido-D-ribose (37)
Preparation 31 was repeated except that 4, 6-O-ethylene-D-glucose in preparation 31 was replaced with D-ribose, and the required raw materials, reagents and preparation method were the same as in preparation 31 to obtain 37.
Preparation example 38: preparation of 1-alpha-azido-D-lyxose (38)
The required raw materials, reagents and preparation methods were the same as in preparation example 31 except that 4, 6-O-ethylene-D-glucose in preparation example 31 was replaced with D-lyxose, to give 38.
Preparation example 39: preparation of 1-beta-azido-N-acetylglucosamine (39)
Preparation 31 was repeated except that 4, 6-O-ethylene-D-glucose in preparation 31 was replaced with N-acetylglucosamine, and the required raw materials, reagents and preparation were changed to those in preparation 31 to obtain 39.
Preparation example 40: preparation of 1-beta-azido-D-galactose (40)
Preparation 31 was repeated except that 4, 6-O-ethylene-D-glucose in preparation 31 was replaced with D-galactose, and the required starting materials, reagents and preparation were the same as in preparation 31 to give 40.
Preparation example 41: preparation of 1-beta-azido-L-fucose (41)
The required raw materials, reagents and preparation method were the same as in preparation example 31 except that 4, 6-O-ethylene-D-glucose in preparation example 31 was replaced with L-fucose, to give 41.
Preparation example 42: preparation of 1-alpha-azido-N-acetamidomanmannose (42)
42 was obtained by following the same procedures as in production example 31 except that 4, 6-O-ethylene-D-glucose in production example 31 was replaced with N-acetamidomanmannose.
Preparation example 43: preparation of 1-alpha-azido-L-lyxose (43)
The required starting materials, reagents and preparation were the same as in preparation example 31 except that 4, 6-O-ethylene-D-glucose in preparation example 31 was replaced with L-lyxose, to give 43.
Preparation example 44: preparation of R-2- (2, 6-dimethoxy-4- (2-methyl-3-phenylbenzyloxy) benzylamino) -propanol (44)
D-aminopropanol (75mg, 1mmol), 2, 6-dimethoxy-4- (2-methyl-3-phenyl benzyloxy) benzaldehyde 1(120mg, 0.33mmol) and sodium triethoxy borohydride (210mg, 1mmol) are mixed and dissolved in 25mL dichloromethane, after full dissolution, the mixture is put in oil bath at 85 ℃ for reaction for 1h, the solvent is removed by reduced pressure distillation, and column chromatography separation (dichloromethane/methanol 100:1, v/v) is carried out to obtain off-white solid R-2- (2, 6-dimethoxy-4- (2-methyl-3-phenyl benzyloxy) benzylamino) -propanol 44.
R-2- (2, 6-dimethoxy-4- (2-methyl-3-phenyl benzyloxy) benzylamino) -propanol (44) high resolution mass spectrum (ESI)+)C26H31NO4Theoretical value of 422.2331, found value of 422.2352[ M + H ]]+。
Preparation example 45: preparation of R-2- (3-chloro-4- (2-methyl-3-phenylphenylmethoxy) benzylamino) -propanol (45)
D-aminopropanol (75mg, 1mmol), 3-chloro-4- (2-methyl-3-phenyl benzyloxy) benzaldehyde 4(111mg, 0.33mmol) and sodium triethoxy borohydride (210mg, 1mmol) are mixed and dissolved in 25mL dichloromethane, after full dissolution, the mixture is put in oil bath at 85 ℃ for reaction for 1h, the solvent is removed by reduced pressure distillation, and column chromatography separation (dichloromethane/methanol 100:1, v/v) is carried out to obtain off-white solid R-2- (3-chloro-4- (2-methyl-3-phenyl benzyloxy) benzylamino) -propanol 45.
R-2- (3-chloro-4- (2-methyl-3-phenyl benzyloxy) benzylamino) -propanol (45) high resolution mass spectrum (ESI)+)C24H26ClNO2Theoretical value of 396.1730, found value of 396.1725[ M + H ]]+。
Preparation example 46: preparation of N- (2- (((2-methoxy- [1, 1' -biphenyl ] -3-yl) methoxy) pyridin-3-yl) methyl) amino) ethyl) acetamide (46)
N-acetylethylenediamine (102mg, 1mmol), 2-methoxy-6- (2-methyl-3-phenylphenylmethoxy) pyridine-3-carbaldehyde 10(119mg, 0.33mmol) was dissolved in DCM (25mL), and reacted at room temperature for 2h after adding acetic acid (2 mL). Sodium triethoxyborohydride (118.7mg,0.56mmol) was added. After 4h of reaction, the solvent was removed by distillation under reduced pressure, and column chromatography (dichloromethane/methanol 100:1 to 30:1, V/V) was performed to give off-white solid N- (2- (((2-methoxy- [1, 1' -biphenyl ] -3-yl) methoxy) pyridin-3-yl) methyl) amino) ethyl) acetamide 46.
N- (2- (((2-methoxy- [1, 1' -biphenyl)) benzene]-3-yl) methoxy) pyridin-3-yl) methyl) amino) ethyl) acetamide (46) high resolution mass spectrometry (ESI)+)C25H29N3O3Theoretical value of 420.2287, found value of 420.2264[ M + H ]]+。
Example (b): product preparation
Example 1
2-aminoethyl-. beta. -D-galactopyranoside (0.42g) was dissolved in 2mL of distilled water and added slowly to a 20mL solution of 2-methoxy-6- (2-methyl-3-phenylphenylmethoxy) pyridine-3-carbaldehyde (0.51g) in DMF and 0.5mL of DIPEA and reacted at 55 ℃ for 4 h. Stopping reaction and naturally coolingAfter cooling to room temperature, TFA was added to adjust pH 6. Sodium triethoxyborohydride (0.63g) was slowly added to the reaction, and after 2h of reaction, sodium triethoxyborohydride (0.32g) was added. After 4 hours of reaction, the solvent was distilled off under reduced pressure and separated by column chromatography (dichloromethane/methanol 100:1, v/v) to give PDL1-017(0.48g) as an off-white solid. High resolution mass spectrometry (ESI +) C29H36N2O8Theoretical value of 541.2550, found value of 541.5746[ M + H ]]+。
1H NMR(400MHz,MeOD-d4):δ(ppm)＝7.73(d,J＝8.1Hz,1H),7.44(t,J＝7.4Hz,2H),7.37(d,J＝7.5Hz,1H),7.31-7.16(m,3H),6.50(d,J＝8.0Hz,1H),5.52(s,2H),4.34(d,J＝7.6Hz,1H),4.22(s,2H),4.12(ddt,J＝12.3,6.7,3.1Hz,1H),4.08(s,3H),3.98(ddd,J＝12.2,6.8,3.3Hz,1H),3.85(d,J＝2.8Hz,1H),3.82-3.71(m,2H),3.58(dd,J＝9.7,7.6Hz,2H),3.52(d,J＝3.3Hz,1H),3.32(s,2H),2.27(s,3H).
13C NMR(400MHz,MeOD-d4):163.75,161.20,143.40,142.91,142.04,135.66,133.97,129.46,128.94,127.95,127.80,126.58,125.04,104.27,103.39,101.91,75.69,73.40,71.04,68.87,66.79,64.15,61.19,53.16,45.33,15.03.
Example 2
The required raw materials, reagents and preparation methods were the same as in example 1 except that 2-methoxy-6- (2-methyl-3-phenylphenylmethoxy) pyridine-3-carbaldehyde in example 1 was replaced with 2, 6-dimethoxy-4- (2-methyl-3-phenylphenylmethoxy) benzaldehyde, whereby PDL1-018 was obtained. High resolution mass spectrometry (ESI +) C31H39NO9Theoretical value of 570.2703, found value of 570.6023[ M + H ]]+。
1H NMR(400MHz,MeOD-d4):δ(ppm)＝7.44(t,J＝7.4Hz,3H),7.38(d,J＝7.2Hz,1H),7.33–7.18(m,4H),6.44(s,2H),5.22(s,2H),4.32(d,J＝7.5Hz,1H),4.27(s,2H),4.09(dd,J＝6.0,2.6Hz,1H),4.00-3.95(m,1H),3.92(s,6H),3.85(d,J＝3.1Hz,1H),3.76(dd,J＝6.0,3.1Hz,2H),3.65-3.54(m,2H),3.51(dd,J＝9.7,3.2Hz,1H),3.21(ddd,J＝10.0,6.4,3.4Hz,2H),2.26(s,3H).
13C NMR(400MHz,MeOD-d4):162.41,159.90,143.02,141.97,135.19,134.22,129.71,128.94,128.01,127.80,126.60,125.15,103.41,99.10,91.11,75.65,73.38,71.03,69.15,68.82,64.06,61.14,55.14,46.61,39.44,14.99.
Example 3
The required raw materials, reagents and preparation method were the same as in example 2 except that 2-aminoethyl- β -D-galactopyranoside in example 2 was replaced with 2-aminoethyl- α -L-rhamnopyranoside, to give PDL 1-019. High resolution mass spectrometry (ESI +) C31H39NO8Theoretical value of 554.2754, found value of 554.2656[ M + H ]]+。
1H NMR(400MHz,MeOD-d4):δ(ppm)＝7.47-7.41(m,3H),7.39-7.34(m,1H),7.30-7.24(m,3H),7.21(dd,J＝7.7,1.6Hz,1H),6.44(s,2H),5.22(s,2H),4.76(d,J＝1.6Hz,1H),4.26(s,2H),3.97(ddd,J＝11.6,5.6,4.1Hz,2H),3.92(s,7H),3.71(td,J＝9.3,8.4,3.7Hz,2H),3.58(dq,J＝9.4,6.2Hz,1H),3.44(t,J＝9.4Hz,1H),3.30-3.18(m,2H),2.26(s,3H),1.30(d,J＝6.2Hz,3H).
13C NMR(400MHz,D2O):162.37,162.02,161.58,159.60,142.12,141.48,135.00,133.85,128.94,127.67,117.90,114.99,100.14,98.76,90.70,71.78,70.20,69.86,68.88,68.63,61.78,55.31,44.88,38.88,16.7,15.41.
Example 4
The required raw materials, reagents and preparation method were the same as example 2 except that 2-aminoethyl- β -D-galactopyranoside in example 2 was replaced with 2-aminoethyl- α -D-mannopyranoside, to obtain PDL 1-020.High resolution mass spectrometry (ESI +) C31H39NO9Theoretical value of 570.2703, found value of 570.3485[ M + H ]]+。
1H NMR(400MHz,MeOD-d4):δ(ppm)＝7.39-7.33(m,1H),7.30-7.20(m,4H),7.47-7.40(m,3H),6.44(s,2H),5.22(s,2H),4.84(d,J＝1.7Hz,1H),4.27(s,2H),4.08–3.98(m,1H),3.92(s,8H),3.79–3.69(m,3H),3.65(t,J＝9.4Hz,1H),3.55(ddd,J＝9.2,6.4,2.2Hz,1H),3.26–3.20(m,2H),2.26(s,3H).
13C NMR(500MHz,D2O):162.28,162.01,161.59,159.63,142.14,141.52,135.02,133.85,128.98,127.69,117.66,115.33,100.07,98.93,90.74,73.02,70.52,69.81,68.59,66.45,61.94,60.72,55.34,45.11,39.06,15.41.
Example 5
The required raw materials, reagents and preparation method were the same as in example 2 except that 2-aminoethyl- β -D-galactopyranoside in example 2 was replaced with 2-aminoethyl-2-acetylamino-2-deoxy- β -D-glucopyranoside, to obtain PDL 1-021. High resolution mass spectrometry (ESI +) C33H42N2O9Theoretical value of 611.2968, found value of 611.3624[ M + H ]]+。
1H NMR(400MHz,MeOD-d4):δ(ppm)＝7.46-7.34(m,4H),7.30-7.20(m,4H),6.43(s,2H),5.22(s,2H),4.45(d,J＝8.4Hz,1H),4.25(d,J＝1.3Hz,2H),4.08(ddd,J＝11.3,7.6,3.4Hz,1H),3.92(s,8H),3.71(ddd,J＝21.7,11.0,7.1Hz,2H),3.47(dd,J＝10.3,8.0Hz,1H),3.39–3.31(m,1H),3.26–3.07(m,2H),2.26(s,3H),2.00(s,3H).
13C NMR(500MHz,D2O):174.32,162.41,162.13,161.61,159.63,142.18,141.49,135.00,133.92,128.97,127.72,117.65,115.33,101.08,98.93,90.80,75.93,73.63,69.89,68.67,64.26,60.67,55.31,45.69,39.13,22.15,15.44.
Example 6
The required raw materials, reagents and preparation method were the same as in example 2 except that 2-aminoethyl- β -D-galactopyranoside in example 2 was replaced with 2-aminoethyl- β -L-fucopyranoside, to obtain PDL 1-022. High resolution mass spectrometry (ESI +) C31H39NO8Theoretical value of 554.2754, found value of 554.2775[ M + H ]]+。
1H NMR(400MHz,MeOD-d4):δ(ppm)＝7.44(dd,J＝8.2,6.7Hz,3H),7.40–7.35(m,1H),7.31–7.21(m,4H),6.44(s,2H),5.23(s,2H),4.30(d,J＝6.9Hz,1H),4.25(s,2H),4.06(m,1H),3.92(s,7H),3.74-3.61(m,2H),3.58-3.46(m,2H),3.22(m,2H),2.26(s,3H),1.27(d,J＝6.4Hz,3H).
13C NMR(500MHz,D2O):164.30,161.88,161.65,160.97,159.04,141.54,140.87,134.41,133.32,128.36,127.10,116.80,114.86,102.06,98.20,90.06,72.13,70.43,70.25,69.72,63.54,54.71,44.91,36.28,30.75,14.82.
Example 7
The required starting materials, reagents and preparation were the same as in example 2 except that 2-aminoethyl- β -D-galactopyranoside from example 2 was replaced with N- (3-aminopropyl) -D-lactosamide, to give PDL 1-023. High resolution mass spectrometry (ESI +) C38H52N2O14Theoretical value of 761.3497, found value of 761.3462[ M + H ]]+。
1H NMR(400MHz,MeOD-d4):δ(ppm)＝7.50-7.18(m,8H),6.43(s,2H),5.22(s,2H),4.51(d,J＝7.6Hz,1H),4.39(d,J＝2.2Hz,1H),4.23(dd,J＝4.4,2.2Hz,1H),4.19(s,2H),3.96(dd,J＝6.6,4.4Hz,2H),3.92(s,6H),3.90-3.68(m,6H),3.64-3.56(m,2H),3.50(ddd,J＝16.6,9.1,4.2Hz,2H),3.05(m,2H),2.26(s,3H),1.95(s,2H).
13C NMR(500MHz,D2O):174.64,162.48,162.20,161.54,159.58,142.18,141.47,134.97,133.89,128.97,127.71,117.63,115.30,103.49,99.34,90.80,81.16,75.41,72.51,72.44,71.44,71.06,70.57,68.62,61.95,61.11,55.36,43.68,38.99,35.60,25.18,15.42.
Example 8
The required starting materials, reagents and preparation were the same as in example 2 except that 2-aminoethyl- β -D-galactopyranoside in example 2 was replaced by 2-aminoethyl- β -D-xylopyranoside, giving PDL 1-024. High resolution mass spectrometry (ESI +) C30H37NO8Theoretical value of 540.6328, found value of 540.2629[ M + H ]]+。
1H NMR(400MHz,MeOD-d4):δ(ppm)＝7.49-7.41(m,3H),7.39-7.34(m,1H),7.31-7.20(m,4H),6.44(s,2H),5.22(s,2H),4.30(d,J＝7.5Hz,1H),4.25(s,2H),4.07(dt,J＝11.9,4.6Hz,1H),3.91(s,6H),3.94-3.83(m,2H),3.51(m,1H),3.37(d,J＝8.9Hz,1H),3.29-3.20(m,4H),2.26(s,3H).
13C NMR(500MHz,D2O):δ(ppm)＝162.37,162.10,161.60,159.63,142.14,141.52,135.03,133.87,128.97,127.68,117.66,115.33,102.89,98.88,90.72,75.62,72.88,69.16,68.59,65.21,64.18,55.31,45.49,38.94,15.42.
Example 9
The required raw materials, reagents and preparation method were the same as example 2 except that 2-aminoethyl- β -D-galactopyranoside in example 2 was replaced with 2-aminoethyl- β -D-glucopyranoside, to obtain PDL 1-025. High resolution mass spectrometry (ESI +) C31H39NO9Theoretical value of 570.2703, found value of 570.2856[ M + H ]]+。
1H NMR(400MHz,MeOD-d4):δ(ppm)＝7.49-7.41(m,3H),7.39-7.34(m,1H),7.31-7.20(m,4H),6.44(s,2H),5.22(s,2H),4.30(d,J＝7.5Hz,1H),4.25(s,2H),4.07(dt,J＝11.9,4.6Hz,1H),3.96–3.84(m,8H),3.51(ddd,J＝10.3,8.7,5.3Hz,1H),3.37(d,J＝8.9Hz,1H),3.29-3.20(m,4H),2.26(s,3H).
13C NMR(500MHz,D2O):δ(ppm)＝161.84,161.61,160.95,159.01,141.55,140.86,134.36,133.32,128.37,127.11,116.82,114.88,101.53,98.23,90.12,75.41,74.97,72.35,68.91,63.71,60.00,54.71,44.97,38.41,14.83.
Example 10
The required starting materials, reagents and preparation were the same as in example 2 except that 2-aminoethyl- β -D-galactopyranoside in example 2 was replaced by N- (3-aminopropyl) -D-glucamide, to give PDL 1-026. High resolution mass spectrometry (ESI +) C32H42N2O9Theoretical value of 599.2968, found value of 599.3211[ M + H ]]+。
1H NMR(400MHz,MeOD-d4):δ(ppm)＝7.48-7.42(m,3H),7.40-7.35(m,1H),7.31-7.21(m,4H),6.43(s,2H),5.23(s,2H),4.27(d,J＝3.3Hz,1H),4.19(s,2H),4.13(t,J＝3.0Hz,1H),3.92(s,6H),3.84-3.61(m,4H),3.47(td,J＝8.5,4.2Hz,1H),3.28(m,1H),3.14–3.00(m,2H),2.27(s,3H),1.96(m,2H).
13C NMR(500MHz,D2O):δ(ppm)＝174.76,162.49,162.22,161.56,159.59,142.17,141.54,135.02,133.87,129.01,127.71,117.65,115.32,99.35,90.75,73.30,72.15,71.13,70.41,62.62,55.32,43.70,39.04,35.59,25.13,15.40.
Example 11
Except that the 2-aminoethyl group in example 1The required starting materials, reagents and preparation were the same as in example 1 except that-beta-D-galactopyranoside was replaced with 2-aminoethyl-alpha-L-rhamnopyranoside to give PDL 1-027. High resolution mass spectrometry (ESI +) C29H36N2O7Theoretical value of 525.2601, found value of 525.2534[ M + H ]]+。
1H NMR(400MHz,MeOD-d4):δ(ppm)＝7.71(d,J＝8.1Hz,1H),7.48–7.39(m,3H),7.38–7.33(m,1H),7.29-7.17(m,4H),6.50(d,J＝8.1Hz,1H),5.51(s,2H),4.77(d,J＝1.7Hz,1H),4.21(s,2H),4.07(s,3H),4.03–3.88(m,2H),3.71(ddd,J＝13.0,8.2,3.7Hz,2H),3.58(dq,J＝9.4,6.2Hz,1H),3.44(t,J＝9.4Hz,1H),3.30(s,1H),2.26(s,3H),1.30(d,J＝6.2Hz,3H).
13C NMR(500MHz,D2O):δ(ppm)＝163.07,162.35,162.07,161.79,160.87,142.03,141.49,135.25,133.69,128.91,127.62,117.61,115.29,103.72,101.65,100.07,71.83,70.20,69.85,68.89,66.53,61.99,53.33,45.29,44.96,16.72,15.41.
Example 12
The required raw materials, reagents and preparation method were the same as in example 1 except that 2-aminoethyl- β -D-galactopyranoside in example 1 was replaced with 2-aminoethyl- α -D-mannopyranoside, to obtain PDL 1-028. High resolution mass spectrometry (ESI +) C29H36N2O8Theoretical value of 541.2550, found value of 541.2846[ M + H ]]+。
1H NMR(400MHz,MeOD-d4):δ(ppm)＝7.72(d,J＝8.2Hz,1H),7.43(dd,J＝8.1,6.6Hz,3H),7.39-7.34(m,1H),7.29-7.17(m,4H),6.51(d,J＝8.0Hz,1H),5.52(s,2H),4.84(d,J＝1.7Hz,1H),4.21(s,2H),4.08(s,3H),3.91-3.87(m,2H),3.81-3.69(m,2H),3.63(t,J＝9.4Hz,1H),3.55(ddd,J＝9.3,6.6,2.2Hz,1H),3.31(s,2H),2.27(s,3H).
13C NMR(500MHz,D2O):δ(ppm)＝163.07,162.41,162.13,160.87,143.60,142.04,141.49,135.23,133.71,128.94,127.63,117.60,115.27,103.82,101.58,100.02,73.00,70.49,69.79,66.48,62.11,60.73,53.35,45.49,45.14,15.40.
Example 13
The required starting materials, reagents and preparation were the same as in example 1 except that 2-aminoethyl- β -D-galactopyranoside in example 1 was replaced with 2-aminoethyl-2-acetylamino-2-deoxy- β -D-glucopyranoside, to give PDL 1-029. High resolution mass spectrometry (ESI +) C31H39N3O8Theoretical value of 582.2815, found value of 582.3145[ M + H ]]+。
1H NMR(400MHz,MeOD-d4):δ(ppm)＝7.73(d,J＝8.1Hz,1H),7.47-7.41(m,3H),7.39-7.33(m,1H),7.31-7.15(m,4H),6.50(d,J＝8.0Hz,1H),5.52(s,2H),4.44(d,J＝8.4Hz,1H),4.26-4.10(m,3H),4.07(s,3H),3.98-3.87(m,2H),3.71(ddd,J＝22.5,11.0,7.1Hz,2H),3.46(dd,J＝10.3,8.1Hz,1H),3.40–3.34(m,2H),3.24(dddd,J＝24.5,13.4,7.3,3.4Hz,2H),2.26(s,3H),2.00(s,3H).
13C NMR(500MHz,D2O):δ(ppm)＝174.32,163.08,162.49,162.21,160.84,143.57,142.06,141.48,135.23,133.71,128.93,127.64,117.60,115.27,103.89,101.63,101.02,75.92,73.64,69.93,66.55,64.35,60.68,55.25,53.35,46.04,45.14,22.19,15.42.
Example 14
The required raw materials, reagents and preparation method were the same as in example 1 except that 2-aminoethyl- β -D-galactopyranoside in example 1 was replaced with 2-aminoethyl- β -L-fucopyranoside, to obtain PDL 1-030. High resolution mass spectrometry (ESI +) C29H36N2O7Theoretical value of 525.2601, found value of 525.2893[ M + H ]]+。
1H NMR(400MHz,MeOD-d4):δ(ppm)＝7.73(d,J＝8.1Hz,1H),7.48-7.40(m,3H),7.39–7.34(m,1H),7.30-7.17(m,4H),6.50(d,J＝8.0Hz,1H),5.52(s,2H),4.31(d,J＝6.8Hz,1H),4.20(s,2H),4.07(s,4H),3.92(ddd,J＝11.9,6.6,3.8Hz,1H),3.69(q,J＝6.5Hz,2H),3.64(d,J＝2.8Hz,1H),3.58–3.46(m,2H),3.33–3.22(m,2H),2.26(s,3H),1.26(d,J＝6.4Hz,3H).
13C NMR(500MHz,D2O):δ(ppm)＝163.08,162.47,162.19,160.89,143.67,142.06,141.49,135.28,133.71,128.94,127.64,117.61,115.28,103.84,102.68,101.63,72.78,71.08,70.86,70.37,66.52,64.25,53.35,45.99,44.89,15.43.
Example 15
The required raw materials, reagents and preparation method were the same as in example 1 except that 2-aminoethyl-. beta. -D-galactopyranoside in example 1 was replaced with N- (3-aminopropyl) -D-lactosamide, to give PDL 1-031. High resolution mass spectrometry (ESI +) C36H49N3O13Theoretical value of 732.3343, found value of 732.3342[ M + H ]]+。
1H NMR(400MHz,MeOD-d4):δ(ppm)＝7.73(d,J＝8.0Hz,1H),7.44(dd,J＝8.0,6.6Hz,3H),7.37(t,J＝7.2Hz,1H),7.30–7.18(m,4H),6.51(d,J＝8.1Hz,1H),5.52(s,2H),4.52(d,J＝7.6Hz,1H),4.41(d,J＝2.3Hz,1H),4.25(dd,J＝4.3,2.4Hz,1H),4.20–4.04(m,5H),3.97(dd,J＝6.6,4.3Hz,1H),3.92-3.70(m,6H),3.63-3.49(m,3H),3.32(s,2H),3.15-3.06(m,2H),2.27(s,3H),1.95(dq,J＝21.9,12.4,9.8Hz,2H).
13C NMR(500MHz,D2O):δ(ppm)＝174.68,163.08,160.82,143.38,142.10,141.45,135.24,133.74,129.42,128.95,127.69,126.34,117.58,115.26,112.93,103.49,81.15,75.40,72.50,72.43,71.43,71.06,70.58,68.63,66.67,61.95,61.12,53.36,45.24,44.15,35.64,25.34,15.41.
Example 16
The required starting materials, reagents and preparation were the same as in example 1 except that 2-aminoethyl- β -D-galactopyranoside in example 1 was replaced by 2-aminoethyl- β -D-xylopyranoside to give PDL 1-032. High resolution mass spectrometry (ESI +) C28H34N2O7Theoretical value of 511.2444, found value of 511.2444[ M + H ]]+。
1H NMR(400MHz,MeOD-d4):δ(ppm)＝7.71(d,J＝8.1Hz,1H),7.43(dd,J＝7.9,6.5Hz,3H),7.38–7.33(m,1H),7.29-7.16(m,4H),6.50(d,J＝8.0Hz,1H),5.51(s,2H),4.32(d,J＝7.5Hz,1H),4.20(s,2H),4.06(s,4H),3.88(ddd,J＝11.4,8.7,5.2Hz,2H),3.52(ddd,J＝10.3,8.6,5.3Hz,1H),3.37(d,J＝8.9Hz,1H),3.31–3.20(m,4H),2.26(s,3H).
13C NMR(500MHz,D2O):δ(ppm)＝163.09,162.40,162.13,160.83,143.55,142.03,141.49,135.24,133.69,128.93,127.60,117.61,115.29,103.77,102.86,101.61,75.62,72.89,69.19,66.52,65.22,64.25,53.30,45.85,45.02,15.41.
Example 17
The required raw materials, reagents and preparation method were the same as in example 1 except that 2-aminoethyl- β -D-galactopyranoside in example 1 was replaced with 2-aminoethyl- β -D-glucopyranoside, to give PDL 1-033. High resolution mass spectrometry (ESI +) C29H36N2O8Theoretical value of 541.2550, found value of 541.5741[ M + H ]]+。
1H NMR(400MHz,MeOD-d4):δ(ppm)＝7.72(d,J＝8.1Hz,1H),7.48-7.41(m,3H),7.39–7.34(m,1H),7.31–7.15(m,4H),6.51(d,J＝8.1Hz,1H),5.52(s,2H),4.37(d,J＝7.8Hz,1H),4.22(s,2H),4.19-4.08(m,1H),4.07(s,3H),4.00-3.88(m,2H),3.66(dd,J＝11.7,6.1Hz,1H),3.42–3.35(m,2H),3.31-3.21(m,4H),2.27(s,3H).
13C NMR(500MHz,D2O):δ(ppm)＝163.10,160.87,143.60,142.08,141.47,135.24,133.76,128.95,127.97,127.66,125.12,117.61,115.28,103.85,102.16,101.58,76.05,75.61,73.00,69.62,66.65,64.50,60.69,53.37,46.07,45.16,15.43.
Example 18
The required starting materials, reagents and preparation were the same as in example 1 except that 2-aminoethyl-. beta. -D-galactopyranoside in example 1 was replaced by N- (3-aminopropyl) -D-glucamide, to give PDL 1-034. High resolution mass spectrometry (ESI +) C30H39N3O8Theoretical value of 570.2815, found value of 570.2708[ M + H ]]+。
1H NMR(400MHz,MeOD-d4):δ(ppm)＝7.72(d,J＝8.1Hz,1H),7.44(td,J＝7.3,6.5,1.7Hz,3H),7.39–7.34(m,1H),7.30-7.17(m,4H),6.49(d,J＝8.1Hz,1H),5.51(s,2H),4.29(d,J＝3.4Hz,1H),4.19–4.04(m,6H),3.85-3.75(m,2H),3.77-3.62(m,2H),3.50(ddd,J＝13.7,8.5,5.0Hz,1H),3.32-3.25(m,1H),3.12(p,J＝5.9Hz,2H),2.26(s,3H),1.95(dt,J＝14.9,7.4Hz,2H).
13C NMR(500MHz,D2O):δ(ppm)＝174.78,163.07,162.44,162.16,160.79,143.38,142.03,141.48,135.20,133.66,128.93,127.60,117.60,115.27,104.15,101.51,73.32,72.14,71.13,70.42,66.51,62.61,53.29,45.18,44.15,35.62,25.29,15.38.
Example 19
Except that 2-aminoethyl-beta-D-galactopyranoside from example 2 was replaced by 2-amino-n-propyl-beta-D-glucopyranosideThe materials, reagents and preparation were the same as in example 2 to obtain PDL 1-051. High resolution mass spectrometry (ESI +) C32H41NO9Theoretical value of 584.2859, found value of 584.4314[ M + H ]]+。
1H NMR(500MHz,MeOD-d4):δ(ppm)＝7.47–7.41(m,3H),7.39–7.35(m,1H),7.31-7.20(m,4H),6.43(s,2H),5.22(d,J＝1.9Hz,2H),4.33(d,J＝7.8Hz,1H),4.21(s,2H),4.07-4.01(m,1H),3.99–3.53(m,10H),3.39(t,J＝8.8Hz,1H),3.32-3.25(m,2H),3.22–3.12(m,2H),2.26(s,3H),2.04(p,J＝6.4Hz,2H).
13C NMR(500MHz,MeOD-d4):δ(ppm)＝162.38,159.90,143.01,141.99,135.22,134.26,129.74,128.97,128.05,127.84,126.62,125.19,102.99,99.28,91.07,76.74,76.69,73.59,70.23,69.15,67.38,61.23,55.15,45.16,39.62,25.50,15.03.
Example 20
The required raw materials, reagents and preparation method were the same as in example 2 except that 2-aminoethyl- β -D-galactopyranoside in example 2 was replaced with 2-amino-n-butyl- β -D-glucopyranoside, to obtain PDL 1-052. High resolution mass spectrometry (ESI +) C33H43NO9Theoretical value of 598.3016, found value of 598.3068[ M + H ]]+。
1H NMR(500MHz,MeOD-d4):δ(ppm)＝7.47-7.41(m,3H),7.39-7.34(m,1H),7.31-7.19(m,4H),6.43(s,2H),5.22(s,2H),4.29(d,J＝7.8Hz,1H),4.20(d,J＝5.1Hz,2H),3.99-3.80(m,8H),3.71–3.35(m,4H),3.31–3.16(m,2H),3.05(q,J＝10.7,9.1Hz,2H),2.26(s,3H),1.91-1.82(m,2H),1.78-1.67(m,2H).
13C NMR(500MHz,MeOD-d4):δ(ppm)＝162.36,159.85,143.01,141.99,135.22,134.25,129.73,128.97,128.05,127.84,126.62,125.18,102.94,99.42,98.75,91.06,76.71,73.72,72.52,70.26,69.14,68.69,66.76,61.30,55.12,39.41,26.19,22.71,15.03.
Example 21
The required raw materials, reagents and preparation method were the same as in example 2 except that 2-aminoethyl- β -D-galactopyranoside in example 2 was replaced with N- (3-aminoethyl) -D-lactosamide, to give PDL 1-053. High resolution mass spectrometry (ESI +) C37H50N2O14Theoretical value of 747.3340, found value of 747.3309[ M + H ]]+。
1H NMR(500MHz,MeOD-d4):δ(ppm)＝7.46-7.38(m,3H),7.38-7.31(m,1H),7.30-7.17(m,4H),6.41(s,2H),5.20(s,2H),4.48(d,J＝7.6Hz,1H),4.35(d,J＝2.4Hz,1H),4.24-4.18(m,1H),4.16(s,2H),3.92(dd,J＝6.8,3.9Hz,1H),3.89(s,7H),3.82-3.73(m,4H),3.68(dd,J＝11.5,4.4Hz,1H),3.60-3.45(m,3H),3.38(dt,J＝13.5,6.8Hz,1H),3.21(q,J＝7.2Hz,3H),2.99(td,J＝7.5,4.2Hz,2H),2.24(s,3H),1.72(q,J＝7.7Hz,2H),1.60(dt,J＝11.3,6.8Hz,2H),1.31(t,J＝7.3Hz,4H).
13C NMR(500MHz,MeOD-d4):δ(ppm)＝174.01,162.36,159.84,143.01,141.99,135.22,134.25,129.73,128.96,128.05,127.83,126.62,125.18,104.35,99.42,91.03,81.72,75.87,73.38,72.61,71.59,71.36,71.11,69.14,68.94,62.30,61.35,55.13,39.43,37.51,26.28,22.48,15.01,7.78.
Example 22
The required raw materials, reagents and preparation method were the same as in example 2 except that 2-aminoethyl-. beta. -D-galactopyranoside in example 2 was replaced with N- (3-aminobutyl) -D-lactosamide, to give PDL 1-054. High resolution mass spectrometry (ESI +) C39H54N2O14Theoretical value of 775.3653, found value of 775.4547[ M + H ]]+。
1H NMR(500MHz,MeOD-d4):δ(ppm)＝7.48–7.41(m,3H),7.33–7.19(m,5H),6.43(d,J＝2.4Hz,2H),5.22(d,J＝2.3Hz,2H),4.89(d,J＝13.1Hz,3H),4.52(d,J＝7.6Hz,1H),4.43(d,J＝2.1Hz,1H),4.25(s,3H),4.00-3.48(m,20H),3.26-3.12(m,6H),2.26(s,3H),1.33(t,J＝7.3Hz,5H).
13C NMR(500MHz,MeOD-d4):δ(ppm)＝175.47,162.44,159.88,143.03,141.99,135.21,134.24,129.73,128.96,128.04,127.83,126.62,125.18,104.33,99.12,91.12,81.75,75.87,73.39,72.58,71.67,71.36,71.18,69.18,68.94,62.35,61.32,55.20,46.51,39.75,35.50,15.01,7.80.
Example 23
The required starting materials, reagents and preparation were the same as in example 2 except that 2-aminoethyl- β -D-galactopyranoside in example 2 was replaced with 2- (β -aminopropionylamino) -D-glucopyranoside to give PDL 1-055. High resolution mass spectrometry (ESI +) C32H40N2O9Theoretical value of 597.2812, found value of 597.4282[ M + H ]]+。
1H NMR(500MHz,MeOD-d4):δ(ppm)＝7.44(tt,J＝7.8,1.4Hz,3H),7.39-7.35(m,1H),7.30-7.20(m,4H),6.46–6.42(m,2H),5.22(s,2H),4.23(d,J＝2.2Hz,2H),3.94–3.63(m,12H),3.50–3.37(m,2H),3.27–3.21(m,2H),2.72(dt,J＝12.4,5.8Hz,2H),2.26(s,3H).
13C NMR(500MHz,MeOD-d4):δ(ppm)＝162.37,159.80,143.01,141.98,135.19,134.23,129.72,128.95,128.02,127.81,126.61,125.17,99.13,91.14,71.73,69.17,63.38,55.16,54.17,39.77,31.65,26.73,25.51,22.33,15.02.
Example 24
Except that 2-aminoethyl-beta-D-galactopyranose is used in example 2The required starting materials, reagents and preparation methods were the same as in example 2 except that the glycoside was replaced with 2- (γ -aminobutyrylamino) -D-glucopyranose, to give PDL 1-056. High resolution mass spectrometry (ESI +) C33H42N2O9Theoretical value of 611.2968, found value of 611.3482[ M + H ]]+。
1H NMR(500MHz,MeOD-d4):δ(ppm)＝7.44(t,J＝7.8Hz,3H),7.40–7.35(m,1H),7.32–7.19(m,4H),6.44–6.41(m,2H),5.22(s,2H),4.19(s,2H),3.94–3.63(m,11H),3.47-3.33(m,2H),3.06(dq,J＝7.7,4.7Hz,2H),2.50–2.42(m,2H),2.26(s,3H),2.04-1.95(m,2H).
13C NMR(500MHz,MeOD-d4):δ(ppm)＝173.66,162.35,159.81,143.02,141.99,135.22,134.25,129.74,128.97,128.05,127.83,126.62,125.18,99.44,91.01,71.70,71.27,71.12,70.78,69.14,61.32,55.12,54.45,39.52,32.64,21.31,15.01.
Example 25
The required raw materials, reagents and preparation method were the same as in example 2 except that 2-aminoethyl- β -D-galactopyranoside in example 2 was replaced with (2R,3R,4R,5S) -6- (2-aminoethylamino) -1,2,3,4, 5-hexanetriol, to obtain PDL 1-057. High resolution mass spectrometry (ESI +) C31H42N2O8Theoretical value of 571.3019, found value of 571.3266[ M + H ]]+。
1H NMR(500MHz,MeOD-d4):δ(ppm)＝7.44-7.40(m,3H),7.37-7.32(m,1H),7.29-7.18(m,4H),6.42(s,2H),5.20(s,2H),4.27(s,2H),4.08(dt,J＝8.3,4.2Hz,1H),3.89(s,6H),3.85(dd,J＝4.7,1.6Hz,1H),3.78(dd,J＝10.7,2.8Hz,1H),3.71-3.63(m,3H),3.45(dd,J＝7.4,5.2Hz,2H),3.39(dd,J＝7.3,5.5Hz,2H),3.29–3.22(m,2H),2.24(s,3H).
13C NMR(500MHz,MeOD-d4):δ(ppm)＝162.66,159.94,143.03,141.98,135.16,134.27,129.76,128.96,128.06,127.83,126.63,125.19,98.58,91.08,71.46,70.75,70.53,69.16,68.36,63.24,55.16,50.13,43.17,42.19,40.19,15.00.
Example 26
The required raw materials, reagents and preparation method were the same as in example 2 except that 2-aminoethyl- β -D-galactopyranoside in example 2 was replaced with (2R,3R,4R,5S) -6- (2-aminopropylamino) -1,2,3,4, 5-hexanetriol, to obtain PDL 1-058. High resolution mass spectrometry (ESI +) C32H44N2O8Theoretical value of 585.3176, found value of 585.6343[ M + H ]]+。
1H NMR(500MHz,MeOD-d4):δ(ppm)＝7.45-7.37(m,3H),7.37-7.30(m,1H),7.29-7.15(m,4H),6.44-6.38(m,2H),5.22-5.16(m,2H),4.20(s,2H),4.08(dp,J＝11.2,4.6Hz,1H),3.91-3.83(m,7H),3.78(dq,J＝10.5,3.1Hz,1H),3.68(pd,J＝11.0,9.3,4.9Hz,3H),3.26-3.03(m,6H),2.26-2.00(m,5H).
13C NMR(500MHz,MeOD-d4):δ(ppm)＝162.46,159.89,143.00,141.98,135.20,134.25,129.74,128.97,128.05,127.83,126.62,125.19,99.07,91.04,71.51,70.82,70.59,69.14,68.34,63.26,55.13,49.71,44.58,43.63,39.62,22.25,15.03.
Example 27
The required raw materials, reagents and preparation methods were the same as in example 7 except that 2, 6-dimethoxy-4- (2-methyl-3-phenylphenylmethoxy) benzaldehyde was replaced with 2, 6-dimethyl-4- (2-methyl-3-phenylphenylmethoxy) benzaldehyde in example 7, to obtain PDL 1-059. High resolution mass spectrometry (ESI +) C38H52N2O12Theoretical value of 729.3598, found value of 719.7785[ M + H ]]+。
Example 28
The required raw materials, reagents and preparation methods were the same as those of example 7 except that 2, 6-dimethoxy-4- (2-methyl-3-phenylphenylmethoxy) benzaldehyde was replaced with 3-fluoro-4- (2-methyl-3-phenylphenylmethoxy) benzaldehyde in example 7, to obtain PDL 1-060. High resolution mass spectrometry (ESI +) C36H47FN2O12Theoretical value of 719.3191, found value of 719.7339[ M + H ]]+。
Example 29
The required raw materials, reagents and preparation method were the same as in example 7 except that 2, 6-dimethoxy-4- (2-methyl-3-phenylphenylmethoxy) benzaldehyde was replaced with 3-chloro-4- (2-methyl-3-phenylphenylmethoxy) benzaldehyde in example 7 to obtain PDL 1-061. High resolution mass spectrometry (ESI +) C36H47ClN2O12Theoretical value of 735.2896, found value of 735.7127[ M + H ]]+。
Example 30
The required starting materials, reagents and preparation were the same as in example 7 except that 2, 6-dimethoxy-4- (2-methyl-3-phenylphenylmethoxy) benzaldehyde was replaced with 3-nitro-4- (2-methyl-3-phenylphenylmethoxy) benzaldehyde in example 7 to obtain PDL 1-062. High resolution mass spectrometry (ESI +) C35H47N3O12Theoretical value of 746.3136, found value of 746.7412[ M + H ]]+。
Example 31
The required raw materials, reagents and preparation methods were the same as in example 7 except that 2, 6-dimethoxy-4- (2-methyl-3-phenylphenylmethoxy) benzaldehyde was replaced with 3- (2-methyl-3-phenylphenylmethoxy) -4-methoxybenzaldehyde in example 7 to obtain PDL 1-063. High resolution mass spectrometry (ESI +) C37H50N2O13Theoretical value of 731.3391, found value of 731.7591[ M + H ]]+。
Example 32
The required starting materials, reagents and preparation were the same as in example 7 except that 2, 6-dimethoxy-4- (2-methyl-3-phenylphenylmethoxy) benzaldehyde was replaced with 3- (2-methyl-3-phenylphenylmethoxy) -4-chlorobenzaldehyde as in example 7 to obtain PDL 1-064. High resolution mass spectrometry (ESI +) C36H47ClN2O12Theoretical value of 735.2896, found value of 735.7139[ M + H ]]+。
Example 33
The required raw materials, reagents and preparation method were the same as those of example 7 except that 2, 6-dimethoxy-4- (2-methyl-3-phenylphenylmethoxy) benzaldehyde was replaced with 2-chloro-5- (2-methyl-3-phenylphenylmethoxy) benzaldehyde in example 7, to obtain PDL 1-065. High resolution mass spectrometry (ESI +) C36H47ClN2O12Theoretical value of 735.2896, found value of 735.7129[ M + H ]]+。
Example 34
Except that 2, 6-dimethoxy-4- (2-methyl-3-phenylbenzyloxy) benzaldehyde in example 7 was replaced with 2- (2-methyl-3-Phenylbenzyloxy) pyridine-5-carbaldehyde and the other necessary raw materials, reagents and preparation method were the same as in example 7, to give PDL 1-066. High resolution mass spectrometry (ESI +) C35H47N3O12Theoretical value of 702.3238, found value of 702.7326[ M + H ]]+。
Example 35
2, 6-dimethoxy-4- (2-methyl-3-phenylphenylmethoxy) 2-propynyl-1-ylamine (0.16g) was dissolved in 4mL of DMF, and 3.2mL of 1-. beta. -azido-4, 6-O-ethylene-D-glucose reaction solution was added. 100mM CuSO is prepared4·5H2O solution (25mg CuSO)4·5H2O in 1mL of distilled water), 200mM BTTAA solution (86mg in 1mL of distilled water), 1M sodium ascorbate solution (198mg in 1mL of distilled water) for use. Mixing CuSO sequentially4·5H2O solution (600. mu.L), BTTAA solution (180. mu.L) and sodium ascorbate solution (800. mu.L) were added to the reaction system and reacted at 37 ℃ overnight. The solvent was dried by spinning and then subjected to column chromatography to obtain PDL 1-087. High resolution mass spectrometry (ESI +) C34H40N4O8Theoretical value of 633.2924, found value of 633.2964[ M + H ]]+。
Example 36
The required raw materials, reagents and preparation methods were the same as in example 35 except that the reaction solution of 1- β -azido-4, 6-O-ethylene-D-glucose was replaced with the reaction solution of 1- β -azido-3-O-methylglucose in example 35, to obtain PDL 1-088. High resolution mass spectrometry (ESI +) C33H40N4O8Theoretical value of 621.2964, found value of 621.2589[ M + H ]]+。
Example 37
The required raw materials, reagents and preparation methods were the same as in example 35 except that the reaction solution of 1- β -azido-4, 6-O-ethylene-D-glucose was replaced with the reaction solution of 1- β -azido-4, 6-O-benzylidene-D-glucose in example 35, to obtain PDL 1-089. High resolution mass spectrometry (ESI +) C39H42N4O8Theoretical value of 695.3081, found value of 695.2445[ M + H ]]+。
Example 38
The required raw materials, reagents and preparation methods were the same as in example 35 except that the reaction solution of 1- β -azido-4, 6-O-ethylene-D-glucose was replaced with the reaction solution of 1- β -azido-D-glucose in example 35, to obtain PDL 1-090. High resolution mass spectrometry (ESI +) C32H32N4O8Theoretical value of 607.2768, found value of 607.2272[ M + H ]]+。
Example 39
The required raw materials, reagents and preparation methods were the same as in example 35 except that the reaction solution of 1- β -azido-4, 6-O-ethylene-D-glucose was replaced with the reaction solution of 1- β -azido-L-arabinose in example 35, to obtain PDL 1-091. High resolution mass spectrometry (ESI +) C31H36N4O7Theoretical value of 577.2662, found value of 577.2188[ M + H ]]+。
Example 40
Except for the 1-. beta. -stack in example 35The required raw materials, reagents and preparation methods were the same as in example 35 except that the reaction solution of N-4, 6-O-ethylene-D-glucose was replaced with the reaction solution of 1- β -azido-D- (+) -xylose, to obtain PDL 1-092. High resolution mass spectrometry (ESI +) C31H36N4O7Theoretical value of 577.2662, found value of 577.2631[ M + H ]]+。
EXAMPLE 41
The same procedures used in example 35 were repeated except for replacing the reaction solution of 1- β -azido-4, 6-O-ethylene-D-glucose in example 35 with the reaction solution of 1- β -azido-D-ribose, thereby obtaining PDL 1-093. High resolution mass spectrometry (ESI +) C31H36N4O7Theoretical value of 577.2662, found value of 577.2679[ M + H ]]+。
Example 42
The required raw materials, reagents and preparation methods were the same as in example 35 except that the reaction solution of 1- β -azido-4, 6-O-ethylene-D-glucose was replaced with the reaction solution of 1- α -azido-D-lyxose in example 35, to obtain PDL 1-094. High resolution mass spectrometry (ESI +) C31H36N4O7Theoretical value of 577.2662, found value of 577.2237[ M + H ]]+。
Example 43
The required raw materials, reagents and preparation methods were the same as in example 35 except that the reaction solution of 1- β -azido-4, 6-O-ethylene-D-glucose was replaced with the reaction solution of 1- β -azido-N-acetylglucosamine in example 35, to obtain PDL 1-095. High resolution qualitySpectrum (ESI +) C34H41N5O8Theoretical value of 648.3033, found value of 648.2338[ M + H ]]+。
Example 44
The required raw materials, reagents and preparation methods were the same as in example 35 except that the reaction solution of 1- β -azido-4, 6-O-ethylene-D-glucose was replaced with the reaction solution of 1- β -azido-D-galactose in example 35, to obtain PDL 1-096. High resolution mass spectrometry (ESI +) C32H38N4O8Theoretical value of 607.2768, found value of 607.2222[ M + H ]]+。
Example 45
The required raw materials, reagents and preparation methods were the same as in example 35 except that the reaction solution of 1- β -azido-4, 6-O-ethylene-D-glucose was replaced with the reaction solution of 1-azido-L-fucose in example 35, to obtain PDL 1-097. High resolution mass spectrometry (ESI +) C31H36N4O8Theoretical value of 591.6848, found value of 591.6246[ M + H ]]+。
Example 46
The required raw materials, reagents and preparation methods were the same as in example 35 except that the reaction solution of 1- β -azido-4, 6-O-ethylene-D-glucose was replaced with the reaction solution of 1- α -azido-N-acetylaminomannose in example 35, to obtain PDL 1-098. High resolution mass spectrometry (ESI +) C34H41N5O8Theoretical value of 648.3033, found value of 648.2859[ M + H ]]+。
Example 47
The required raw materials, reagents and preparation methods were the same as in example 35 except that the reaction solution of 1- β -azido-4, 6-O-ethylene-D-glucose was replaced with the reaction solution of 1- α -azido-L-lyxose in example 35, to obtain PDL 1-099. High resolution mass spectrometry (ESI +) C31H36N4O7Theoretical value of 577.2662, found value of 577.2090[ M + H ]]+。
Example 48
2-methoxy-6- (2-methyl-3-phenylphenylmethoxy) pyridin-3-propynyl-1-ylamine (0.15g) was dissolved in 4mL of DMF, and 3.2mL of 1-. beta. -azido-4, 6-O-ethylene-D-glucose reaction solution was added. Prepared with 100mM CuSO4·5H2O solution (25mg CuSO)4·5H2O in 1mL of distilled water), 200mM BTTAA solution (86mg in 1mL of distilled water), 1M sodium ascorbate solution (198mg in 1mL of distilled water) for use. Mixing CuSO sequentially4·5H2O solution (600. mu.L), BTTAA solution (180. mu.L) and sodium ascorbate solution (800. mu.L) were added to the reaction system and reacted at 37 ℃ overnight. The solvent was dried by spinning and then subjected to column chromatography to obtain PDL 1-100. High resolution mass spectrometry (ESI +) C32H37N5O7Theoretical value of 604.2771, found value of 604.2713[ M + H ]]+。
Example 49
The same procedures as in example 4 were repeated except that the reaction solution of 1-. beta. -azido-4, 6-O-ethylene-D-glucose in example 48 was replaced with the reaction solution of 3-O-methyl-glucose8, obtaining PDL 1-101. High resolution mass spectrometry (ESI +) C31H37N5O7Theoretical value of 592.2771, found value of 592.2798[ M + H ]]+。
Example 50
PDL1-102 was obtained in the same manner as in example 48 except that the reaction solution of 1- β -azido-4, 6-O-ethylene-D-glucose was replaced with 1- β -azido-4, 6-O-benzylidene-D-glucose in example 48. High resolution mass spectrometry (ESI +) C37H39N5O7Theoretical value of 666.2927, found value of 666.2961[ M + H ]]+。
Example 51
PDL1-103 was obtained in the same manner as in example 48 except that the reaction solution of 1- β -azido-4, 6-O-ethylene-D-glucose was replaced with the reaction solution of 1- β -azido-D-glucose in example 48. High resolution mass spectrometry (ESI +) C30H35N5O7Theoretical value of 578.2614, found value of 578.2713[ M + H ]]+。
Example 52
The required raw materials, reagents and preparation methods were the same as in example 48 except that the reaction solution of 1- β -azido-4, 6-O-ethylene-D-glucose was replaced with the reaction solution of 1- β -azido-D- (+) -xylose in example 48, to obtain PDL 1-104. High resolution mass spectrometry (ESI +) C29H33N5O6Theoretical value of 548.2509, found value of 548.2535[ M + H ]]+。
Example 53
The required raw materials, reagents and preparation methods were the same as in example 48 except that the reaction solution of 1- β -azido-4, 6-O-ethylene-D-glucose was replaced with the reaction solution of 1- β -azido-D-ribose in example 48, to obtain PDL 1-105. High resolution mass spectrometry (ESI +) C29H33N5O6Theoretical value of 548.2509, found value of 548.2582[ M + H ]]+。
Example 54
PDL1-106 was obtained in the same manner as in example 48 except that the reaction solution of 1- β -azido-4, 6-O-ethylene-D-glucose was replaced with the reaction solution of 1- α -azido-D-lyxose in example 48. High resolution mass spectrometry (ESI +) C29H33N5O6Theoretical value of 548.2509, found value of 548.2562[ M + H ]]+。
Example 55
The required raw materials, reagents and preparation methods were the same as in example 48 except that the reaction solution of 1- β -azido-4, 6-O-ethylene-D-glucose was replaced with the reaction solution of 1- β -azido-N-acetylglucosamine in example 48, to obtain PDL 1-107. High resolution mass spectrometry (ESI +) C32H38N6O7Theoretical value of 619.2880, found value of 619.2913[ M + H ]]+。
Example 56
PDL1-108 was obtained in the same manner as in example 48 except that the reaction solution of 1- β -azido-4, 6-O-ethylene-D-glucose was replaced with the reaction solution of 1- β -azido-D-galactose in example 48, and the required raw materials, reagents and preparation method were changed. High resolution mass spectrometry (ESI +) C30H35N5O7Theoretical value of 578.2614, found value of 578.2588[ M + H ]]+。
Example 57
The required raw materials, reagents and preparation methods were the same as in example 48 except that the reaction solution of 1- β -azido-4, 6-O-ethylene-D-glucose was replaced with the reaction solution of 1- β -azido-L-fucose in example 48, to obtain PDL 1-109. High resolution mass spectrometry (ESI +) C29H33N5O7Theoretical value of 562.2665, found value of 562.2246[ M + H ]]+。
Example 58
The required raw materials, reagents and preparation methods were the same as in example 48 except that the reaction solution of 1- β -azido-4, 6-O-ethylene-D-glucose was replaced with the reaction solution of 1- α -azido-N-acetylaminomannose in example 48, to obtain PDL 1-110. High resolution mass spectrometry (ESI +) C32H38N6O7Theoretical value of 619.2880, found value of 619.2275[ M + H ]]+。
Example 59
Dissolving 3- (2-methyl-3-phenyl benzyloxy) -4-propynyl-1-amine (0.15g) in 4mL of DMF, and adding 1-beta-azido-3-O-methyl-D-glucose3.2mL of the reaction solution. Prepared with 100mM CuSO4·5H2O solution (25mg CuSO)4·5H2O in 1mL of distilled water), 200mM BTTAA solution (86mg in 1mL of distilled water), 1M sodium ascorbate solution (198mg in 1mL of distilled water) for use. Mixing CuSO sequentially4·5H2O solution (600. mu.L), BTTAA solution (180. mu.L) and sodium ascorbate solution (800. mu.L) were added to the reaction system and reacted at 37 ℃ overnight. The solvent was dried by spinning and then subjected to column chromatography to obtain PDL 1-111. High resolution mass spectrometry (ESI +) C32H38N4O7Theoretical value of 591.2818, found value of 591.2634[ M + H ]]+。
Example 60
The required raw materials, reagents and preparation methods were the same as in example 59 except that the reaction solution of 1- β -azido-3-O-methyl-D-glucose was replaced with the reaction solution of 1- β -azido-4, 6-O-benzylidene-D-glucose in example 59, to obtain PDL 1-112. High resolution mass spectrometry (ESI +) C38H40N4O7Theoretical value of 665.2975, found value of 665.2928[ M + H ]]+。
Example 61
The required raw materials, reagents and preparation methods were the same as in example 59 except that the 1- β -azido-3-O-methyl-D-glucose reaction solution in example 59 was replaced with a 1- β -D-glucose reaction solution, to obtain PDL 1-113. High resolution mass spectrometry (ESI +) C31H36N4O7Theoretical value of 577.2662, found value of 577.2655[ M + H ]]+。
Example 62
The required raw materials, reagents and preparation methods were the same as in example 59 except that the reaction solution of 1- β -azido-3-O-methyl-D-glucose was replaced with the reaction solution of 1- β -azido-L-arabinose in example 59, to obtain PDLs 1-114. High resolution mass spectrometry (ESI +) C30H34N4O6Theoretical value of 547.2826, found value of 547.2327[ M + H ]]+。
Example 63
The required raw materials, reagents and preparation methods were the same as in example 59 except that the reaction solution of 1- β -azido-3-O-methyl-D-glucose was replaced with the reaction solution of 1- β -azido-D- (+) -xylose in example 59, to obtain PDL 1-115. High resolution mass spectrometry (ESI +) C30H34N4O6Theoretical value of 547.2826, found value of 547.2854[ M + H ]]+。
Example 64
The required raw materials, reagents and preparation methods were the same as in example 59 except that the reaction solution of 1- β -azido-3-O-methyl-D-glucose was replaced with the reaction solution of 1- β -azido-D-ribose in example 59, to obtain PDL 1-116. High resolution mass spectrometry (ESI +) C30H34N4O6Theoretical value of 547.2826, found value of 547.2184[ M + H ]]+。
Example 65
Except that the reaction solution of 1-beta-azido-3-O-methyl-D-glucose in example 59 was replaced with the reaction solution of 1-alpha-azido-D-lyxoseThe starting materials, reagents and preparation were carried out in the same manner as in example 59 to obtain PDL 1-117. High resolution mass spectrometry (ESI +) C30H34N4O6Theoretical value of 547.2826, found value of 547.2645[ M + H ]]+。
Example 66
The required raw materials, reagents and preparation methods were the same as in example 59 except that the reaction solution of 1- β -azido-3-O-methyl-D-glucose was replaced with the reaction solution of 1- β -azido-N-acetylglucosamine in example 59, to give PDL 1-118. High resolution mass spectrometry (ESI +) C33H39N5O7Theoretical value of 618.2927, found value of 618.2960[ M + H ]]+。
Example 67
The required raw materials, reagents and preparation methods were the same as in example 59 except that the reaction solution of 1- β -azido-3-O-methyl-D-glucose was replaced with the reaction solution of 1- β -azido-D-galactose in example 59, to obtain PDL 1-119. High resolution mass spectrometry (ESI +) C31H36N4O7Theoretical value of 577.2662, found value of 577.2213[ M + H ]]+。
Example 68
The required raw materials, reagents and preparation methods were the same as in example 59 except that the reaction solution of 1- β -azido-3-O-methyl-D-glucose was replaced with the reaction solution of 1- β -azido-L-fucose in example 59, to obtain PDL 1-120. High resolution mass spectrometry (ESI +) C30H34N4O7Theoretical value of 561.2173, found value of 561.2322[ M + H ]]+。
Example 69
The procedure of example 59 was repeated except for using 1-. beta. -azido-3-O-methyl-D-glucose reaction solution in example 59 instead of 1-. alpha. -azido-L-lyxose reaction solution, to obtain PDL 1-121. High resolution mass spectrometry (ESI +) C29H33N5O6Theoretical value of 548.2509, found value of 548.2576[ M + H ]]+。
And (3) activity test:
experimental example 1: test for inhibiting PD-1/PD-L1 protein interaction activity
The 27 compounds of the invention were tested for their in vitro activity in inhibiting the interaction of PD-1/PD-L1 protein. Compound concentration dilutions were performed according to the 2006 CLSI (clinical laboratory standards institute) dilution method and tested for activity in inhibiting PD-1/PD-L1 protein interactions. The results are shown in table 1 below.
The test method comprises the following steps: the test compound was dissolved in a certain volume of DMSO to prepare a solution having a concentration of 10mM as an initial concentration, and then diluted 10-fold with distilled water. Adding 2 μ L of compounds with different concentrations into 384-well plates mixed with 4 μ L of Tag1-PD-L1 protein and 4 μ L of Tag2-PD1 protein, mixing, standing for 15 min, adding 5 μ L of anti-Tag1-Eu, mixing3+And 5. mu.L of anti-Tag2-XL665, while using the compound of preparation example 45 as a control, and left to stand at room temperature for 2 hours before detection in a multifunctional microplate reader (Synergy H4 from BioTek).
TABLE 1 inhibition of PD-1/PD-L1 protein interaction Activity results for the inventive Compounds
The test result of the activity for inhibiting the interaction of the PD-1/PD-L1 protein shows that the compound has certain activity for inhibiting the interaction of the PD-1/PD-L1 protein. Meanwhile, compared with the similar preparation example 45 without the sugar structure, the sugar structure-containing compound provided by the invention has stronger activity of inhibiting the interaction of PD-1/PD-L1 protein.
Experimental example 2: cytotoxicity test
The compounds of the invention were tested for toxicity at the cellular level. Compound concentration dilutions were performed and toxicity tested according to the 2006 CLSI (clinical laboratory standards institute) dilution method. The results are shown in table 2 below.
The test method comprises the following steps: jurkat clone E6-1 cells were inoculated into a 96-well plate, the test compound was dissolved in a certain volume of DMSO to prepare a 10mM solution, which was used as the initial concentration, and then diluted 3-fold with the cell culture medium, 10. mu.L of different concentrations of the compound was added to the cells, placed at 37 ℃ and 5% CO2After 48 hours of incubation, MTT was added, incubated at 37 ℃ for 4 hours, and 100. mu.L of the triple solution was added overnight. OD values were read on a microplate reader at 570nm and 690nm wavelengths. The compounds of Tecntriq, preparations 44 to 46 were used as controls.
TABLE 2 inhibitory Activity of the Compounds of the present invention on cell proliferation in vitro culture of Jurkat clone E6-1
The test results of the compounds show that compared with the similar preparation examples 44, 45 and 46 without sugar structures, the sugar structure-containing compound provided by the invention has the advantages that the cytotoxicity is obviously reduced, the high-concentration compound has no influence on the cell growth, and the compound has better safety.
Experimental example 3: PD-L1 protein degradation assay
The compounds of the invention were tested for cellular level PD-L1 protein degradation. Compound concentration dilutions were performed according to the 2006 CLSI (clinical laboratory standards institute) dilution method and tested for their activity to degrade PD-L1.
The test method comprises the following steps: the degradation effect of the compound on PD-L1 protein is detected by using a Western Blot method. MC-38/hPD-L1 cells were seeded in a six-well plate, test compounds were dissolved in a volume of DMSO to prepare a 10mM solution as an initial concentration, and then diluted with cell culture medium, and after 72 hours of action at various concentrations, the cells were lysed with 1 XSDS gel loading buffer (50mM Tris-HCl (pH 6.8), 100mM DTT, 2% SDS, 10% glycerol, 0.1% bromophenol blue). Heating and denaturing the cell lysate in a boiling water bath, performing SDS-PAGE electrophoresis, transferring the protein to a PVDF membrane by a wet transfer system after the electrophoresis is finished, sealing the PVDF membrane in a sealing solution (5% skimmed milk powder is diluted in TBS/T) at room temperature for 1 hour, and then performing I and II anti-reaction; after washing, the film was developed with ECL reagent. The results are shown in FIG. 1.
FIG. 1 shows that PDL-028 has a certain degradation effect on MC-38/H-11 expressed PD-L1 protein at a concentration of 10. mu.M.
Experimental example 4: activity test of Compounds on the Effect of IFN- γ secretion in Mixed Lymphocyte Reaction (MLR) reactions
The compounds of the invention were subjected to Dendritic Cell (DC) and CD4+Activity test of the Effect of IFN- γ secretion in T-cell Mixed Lymphocyte Reaction (MLR). Dilute of compound concentration and assay for Dendritic Cells (DCs) and CD4 according to the 2006 CLSI (clinical laboratory standards institute) dilution method+Role of IFN- γ secretion in T cell Mixed Lymphocyte Reaction (MLR). The results are shown in table 3 below.
Human Peripheral Blood Mononuclear Cells (PBMCs) were purchased from the shanghai hospital, shanghai, and cultured in RPMI1640 medium containing 10% Fetal Bovine Serum (FBS).
The experimental method comprises the following steps:
1. CD14+ monocytes were sorted from donor PBMC using a CD14+ monocyte sorting kit, and then induced with GM-CSF and rh IL-4 to differentiate into mature DC cells.
2. CD4 was sorted from PBMCs using CD4+ T cell magnetic bead sorting kit+T cells.
3. Will CD4+Mixing T cells and DC cells, and addingDifferent concentrations of the compound were added and incubated at 37 ℃ for 5 days.
4. And detecting the content of IFN-gamma in the supernatant by using an IFN-gamma detection kit.
TABLE 3 IFN-gamma secretion stimulating Activity of the Compounds of the invention in response to MLR
The activity test result of the compound on the influence of IFN-gamma secretion in MLR reaction shows that most compounds effectively promote the secretion of IFN-gamma. Meanwhile, compared with the preparation examples 44 and 46 of the same kind without sugar structure, the sugar structure-containing compound provided by the invention has stronger activity of promoting the secretion of IFN-gamma.
Experimental example 5: in vivo tumor inhibition Activity test in mice
Compounds PDL1-025, PDL1-029 and PDL1-031 of the present invention were tested for anti-tumor activity in mice.
BALB/cA-nude mice, 5-6 weeks old, female, SPF grade, purchased from Shanghai Ling Biotech, Inc.; growing license number: SCXK (Shanghai) 2013-0018; animal certification number 2013001816884; a breeding environment: SPF grade.
The experimental method comprises the following steps: nude mice are inoculated with MC-38/hPD-L1 cells subcutaneously until the tumor grows to 100mm3Thereafter, animals were randomly grouped. Dosing according to the experimental protocol, the solvent group was intragastrically perfused with the same volume of normal saline; tumor size and mouse body weight were recorded every 3-4 days. The results are shown in table 4 and fig. 2.
TABLE 4 test results of antitumor Activity of Compounds of the present invention in mice
The results show that PDL1-025 has a tumor inhibition rate of 21.9% on colon cancer MC-38/hPD-L1, and can effectively inhibit the growth of tumors.
Experimental example 6: analysis of Tumor Infiltrating Lymphocyte (TIL) proportion in mice
The compounds PDL1-025 and PDL1-031 of the present invention were analyzed for the proportion of tumor-infiltrating lymphocytes in mice.
The experimental method comprises the following steps: mice at the end point of the experiment of experiment example 5 were taken, the mice were sacrificed by dislocation of the cervical vertebrae, tumors were taken out with scissors and forceps, cut into pieces with scissors and transferred to a 50ml sterile centrifuge tube, collagenase and hyaluronidase were added, shaken on a shaker at 37 ℃ for 1 hour, a certain proportion of mouse lymphocyte separation fluid was added for centrifugation, TIL was extracted, a control group (negative control), a tecentriq group and an experiment group were set, an antibody was added for staining, and then flow (BD Accuri C6) detection was performed. The results are shown in FIG. 3. FIG. 3 shows that PDL1-025 and PDL1-031 have a promoting effect on TIL and improved CD3+CD4+T cells and CD3+CD8+Proportion of T cells.
Claims (10)
1. A compound of formula I or a pharmaceutically acceptable salt thereof:
wherein:
x is selected from N or C-R5，
R5Selected from H, halogen, nitro, cyano, selected from C1-C6Amino substituted by 1 or 2 substituents in a linear or branched alkyl radical, C1-C5Straight or branched alkyl, C1-C5Straight or branched alkoxy or C1-C5A linear or branched alkylthio group; preferably, R5Selected from H, halogen, nitro, C1-C5Straight or branched alkyl or C1-C5A linear or branched alkoxy group; more preferably, R5Selected from H, methyl, ethyl, propyl, fluorine, chlorine, nitro, methoxy, ethoxy or propoxy;
R1and R2Each independently selected from H, halogen, nitro, cyano, from C1-C5Of straight-chain or branched alkyl groupsAmino substituted by 1 or 2 substituents, C1-C5Straight or branched alkyl, C1-C5Straight or branched alkoxy, C1-C5Straight or branched alkylthio or-CH2NH–(CH2)m-Y-Z, and R1And R2At least one is selected from-CH2NH–(CH2)m-Y-Z; or, R1And R2Each independently selected from H, halogen, C1-C5Straight or branched alkyl, C1-C5Straight or branched alkoxy or-CH2NH–(CH2)m-Y-Z, and R1And R2At least one is selected from-CH2NH–(CH2)m-Y-Z; or, R1And R2Each independently selected from H, methyl, ethyl, methoxy, ethoxy, fluorine, chlorine or-CH2NH–(CH2)m-Y-Z, and R1And R2At least one is selected from-CH2NH–(CH2)m-Y-Z wherein
m is an integer between 1 and 5, preferably 1,2,3 or 4;
y is absent or is-S-, -O-, -NR6–、–C(O)NH–、–NHC(O)–、–(CH2)q-or a triazole, which is a nitrogen-containing compound,
wherein:
q is an integer between 1 and 2;
R6selected from H, C1-C5A linear or branched alkyl group;
preferably, Y is-O-, -NH-, -C (O) NH-, -NHC (O) -,
z is selected from a modified or unmodified monosaccharide or disaccharide residue;
R3and R4Each independently selected from H, halogen, nitro, cyano, from C1-C6Amino substituted by 1 or 2 substituents in a linear or branched alkyl radical, C1-C5Straight or branched alkyl, C1-C5Straight or branched alkoxy or C1-C5A linear or branched alkylthio group; preferably, R3And R4Each independently selected from H, halogen, nitro, C1-C5Straight or branched alkyl or C1-C5A linear or branched alkoxy group; more preferably, R3And R4Each independently selected from H, methyl, ethyl, propyl, fluoro, chloro, nitro, methoxy, ethoxy or propoxy.
2. A compound of formula I or a pharmaceutically acceptable salt thereof according to claim 1, wherein,
in the general formula I, the compound of formula I,
x is selected from N or C-R5，
Wherein, when X is N,
R1is-CH2NH–(CH2)m-Y-Z wherein
m is an integer between 1 and 5, preferably 1,2,3 or 4;
y is-O-, -NH-, -C (O) NH-, -NHC (O) -, or triazole, specifically, Y is-O-, -NH-, -C (O) NH-, -NHC (O) -, or,
Z is selected from a modified or unmodified monosaccharide or disaccharide residue;
R2、R3and R4Each independently selected from H, halogen, nitro, cyano, C1-C5 straight or branched chain alkyl or C1-C5 straight or branched chain alkoxy, preferably, R2、R3And R4Each independently selected from H, methyl, ethyl, propyl, fluoro, chloro, nitro, methoxy, ethoxy or propoxy,
wherein, when X is C-R5When the temperature of the water is higher than the set temperature,
R1、R2and R4Each independently selected from H, halogen, nitro, cyano, C1-C5 straight chain or branched chain alkyl, C1-C5 straight or branched chain alkoxy, or-CH2NH–(CH2)m-Y-Z, and R1、R2And R4At least one of them is selected from-CH2NH–(CH2)m-Y-Z; or, R1、R2And R4Each independently selected from H, halogen, C1-C5Straight or branched alkyl, C1-C5Straight or branched alkoxy or-CH2NH–(CH2)m-Y-Z, and R1、R2And R4At least one is selected from-CH2NH–(CH2)m-Y-Z; or, R1、R2And R4Each independently selected from H, methyl, ethyl, methoxy, ethoxy, fluorine, chlorine or-CH2NH–(CH2)m-Y-Z, and R1、R2And R4At least one is selected from-CH2NH–(CH2)m-Y-Z wherein
m is an integer between 1 and 5, preferably 1,2,3 or 4;
y is-O-, -NH-, -C (O) NH-, -NHC (O) -, or triazole; specifically, Y is-O-, -NH-, -C (O) NH-, -NHC (O) -,
z is selected from a modified or unmodified monosaccharide or disaccharide residue;
R3and R5Each independently selected from H, halogen, nitro, cyano, C1-C5 straight or branched chain alkyl or C1-C5 straight or branched chain alkoxy; preferably, R3And R5Each independently selected from H, methyl, ethyl, propyl, fluoro, chloro, nitro, methoxy, ethoxy or propoxy.
3. A compound of formula I or a pharmaceutically acceptable salt thereof according to claim 1, wherein,
the mono-or disaccharide residue is selected from one of the following residues:
4. a compound of formula I or a pharmaceutically acceptable salt thereof according to claim 1, wherein,
in the general formula I, the compound of formula I,
x is selected from N or C-R5，
R1is-CH2NH–(CH2)m-Y-Z, wherein m, Y and Z are each as defined in claim 1;
R2and R4Each independently selected from H, halogen, C1-C5 straight or branched chain alkyl or C1-C5 straight or branched chain alkoxy; preferably, R2And R4Each independently selected from H, methyl, ethyl, propyl, methoxy, ethoxy or propoxy; more preferably, R2And R4Each independently selected from H, methyl or methoxy;
R3and R5Each independently selected from H, halogen, nitro, C1-C5 straight or branched chain alkyl or C1-C5 straight or branched chain alkoxy; preferably, R3And R5Each independently selected from H, halogen, nitro, methyl, ethyl, propyl, methoxy, ethoxy or propoxy, more preferably R3And R5Each independently selected from H, fluoro, chloro, nitro, methyl or methoxy.
5. A compound of formula I or a pharmaceutically acceptable salt thereof according to claim 1, wherein,
in the general formula I, the compound of formula I,
x is C-R5，
R1Selected from H or halogen;
R2is-CH2NH–(CH2)m-Y-Z, wherein m, Y and Z are each as defined in claim 1; r4Is H; r3And R5Each independently selected fromH. Halogen, nitro, C1-C5 straight or branched chain alkyl or C1-C5 straight or branched chain alkoxy; preferably, R3And R5Each independently selected from H, halogen, nitro, methyl, ethyl, propyl, methoxy, ethoxy or propoxy, more preferably R3And R5Each independently selected from H, fluoro, chloro, nitro, methyl or methoxy.
6. A compound of formula I according to claim 1, or a pharmaceutically acceptable salt thereof, wherein the compound of formula I is selected from one of the following compounds:
7. a process for the preparation of a compound according to claim 1, said process comprising the steps of:
as in the above reaction formula, the compound of the general formula d and the amine compound containing monosaccharide or disaccharide residue are subjected to reductive amination reaction to obtain the compound of the general formula I, wherein the reductive amination reaction is carried out in the presence of alkali and solvent under the action of a reducing agent,
wherein, in the above general formula d, R1a、R2aAnd R4aOne is an aldehyde group, and when R is1aWhen it is an aldehyde group, R2aAnd R4aAre as defined in claim 1 for R2And R4When R is defined as2aWhen it is an aldehyde group, R1aAnd R4aAre as defined in claim 1 for R1And R4When R is defined as4aWhen it is an aldehyde group, R1aAnd R2aAre as defined in claim 1 for R1And R2The definition of (1); in the compounds of the formula I, X, R1，R2，R3And R4Are as defined in claim 1, respectively,
preferably, in the reductive amination reaction,
the solvent includes halogenated hydrocarbon solvents such as dichloromethane, 1, 2-dichloroethane and chloroform, aromatic hydrocarbon solvents such as benzene and toluene, aprotic solvents such as N, N-dimethylformamide, dimethylsulfoxide and hexamethylphosphoramide, ether solvents such as tetrahydrofuran, diethyl ether and 1, 4-dioxane, protic solvents such as water, methanol and ethanol, or a mixture of these solvents;
the base includes triethylamine, pyridine, N-diisopropylethylamine, 4-dimethylaminopyridine, 1, 8-diazabicyclo [5.4.0] -7-undecene, and 1,2,2,6, 6-pentamethylpiperidine; and
the reducing agent includes sodium borohydride, sodium cyanoborohydride, sodium triethoxyborohydride and sodium triacetoxyborohydride.
8. A pharmaceutical composition comprising as an active ingredient a therapeutically effective amount of one or more selected from the compounds of any one of claims 1-6 or pharmaceutically acceptable salts thereof, and optionally a pharmaceutically acceptable carrier, excipient, adjuvant and/or diluent.
9. Use of a compound of any one of claims 1-6, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of claim 8, in the preparation of a PD-L1 inhibitor.
10. Use of a compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 8, for the manufacture of a medicament for the treatment of a neoplastic disease and/or a medicament having an immunomodulatory effect.
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