CN113527386B - Intermediate of vaccine adjuvant MPLA, synthesis and application - Google Patents
Intermediate of vaccine adjuvant MPLA, synthesis and application Download PDFInfo
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- CN113527386B CN113527386B CN202010306826.6A CN202010306826A CN113527386B CN 113527386 B CN113527386 B CN 113527386B CN 202010306826 A CN202010306826 A CN 202010306826A CN 113527386 B CN113527386 B CN 113527386B
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- GWAOFSOXGNJTAQ-UHFFFAOYSA-N benzyl phosphono hydrogen phosphate Chemical compound OP(O)(=O)OP(O)(=O)OCC1=CC=CC=C1 GWAOFSOXGNJTAQ-UHFFFAOYSA-N 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 125000002091 cationic group Chemical group 0.000 description 1
- 210000002421 cell wall Anatomy 0.000 description 1
- 230000007969 cellular immunity Effects 0.000 description 1
- 229940032077 cervical cancer vaccine Drugs 0.000 description 1
- 229910052801 chlorine Inorganic materials 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 229940125782 compound 2 Drugs 0.000 description 1
- 229940126214 compound 3 Drugs 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
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- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- NQGIJDNPUZEBRU-UHFFFAOYSA-N dodecanoyl chloride Chemical compound CCCCCCCCCCCC(Cl)=O NQGIJDNPUZEBRU-UHFFFAOYSA-N 0.000 description 1
- 239000003602 elastase inhibitor Substances 0.000 description 1
- 238000000105 evaporative light scattering detection Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 235000019387 fatty acid methyl ester Nutrition 0.000 description 1
- IRXSLJNXXZKURP-UHFFFAOYSA-N fluorenylmethyloxycarbonyl chloride Chemical compound C1=CC=C2C(COC(=O)Cl)C3=CC=CC=C3C2=C1 IRXSLJNXXZKURP-UHFFFAOYSA-N 0.000 description 1
- SPSXSWRZQFPVTJ-ZQQKUFEYSA-N hepatitis b vaccine Chemical compound C([C@H](NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CO)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC=1C2=CC=CC=C2NC=1)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](N)CCSC)C(=O)N[C@@H](CC1N=CN=C1)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C(C)C)C(=O)OC(=O)CNC(=O)CNC(=O)[C@H](C)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CC(C)C)NC(=O)CNC(=O)[C@@H](N)CCCNC(N)=N)C1=CC=CC=C1 SPSXSWRZQFPVTJ-ZQQKUFEYSA-N 0.000 description 1
- 229940124736 hepatitis-B vaccine Drugs 0.000 description 1
- 229930195733 hydrocarbon Natural products 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 150000002466 imines Chemical class 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 230000001571 immunoadjuvant effect Effects 0.000 description 1
- 230000002163 immunogen Effects 0.000 description 1
- 239000000568 immunological adjuvant Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 150000007529 inorganic bases Chemical class 0.000 description 1
- 239000003456 ion exchange resin Substances 0.000 description 1
- 229920003303 ion-exchange polymer Polymers 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 238000011031 large-scale manufacturing process Methods 0.000 description 1
- 239000010410 layer Substances 0.000 description 1
- 229920006008 lipopolysaccharide Polymers 0.000 description 1
- 239000002502 liposome Substances 0.000 description 1
- 229940124590 live attenuated vaccine Drugs 0.000 description 1
- 229940023012 live-attenuated vaccine Drugs 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 229940124735 malaria vaccine Drugs 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000012046 mixed solvent Substances 0.000 description 1
- JDAKPBBWYIWXJF-UHFFFAOYSA-N n,n-diethyl-1,5-dihydro-2,4,3-benzodioxaphosphepin-3-amine Chemical compound C1OP(N(CC)CC)OCC2=CC=CC=C21 JDAKPBBWYIWXJF-UHFFFAOYSA-N 0.000 description 1
- 150000002825 nitriles Chemical class 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 229910052763 palladium Inorganic materials 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 239000012071 phase Substances 0.000 description 1
- 230000007112 pro inflammatory response Effects 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 102000005962 receptors Human genes 0.000 description 1
- 108020003175 receptors Proteins 0.000 description 1
- 239000004627 regenerated cellulose Substances 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- YAYGSLOSTXKUBW-UHFFFAOYSA-N ruthenium(2+) Chemical compound [Ru+2] YAYGSLOSTXKUBW-UHFFFAOYSA-N 0.000 description 1
- 239000004576 sand Substances 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000007086 side reaction Methods 0.000 description 1
- 230000008961 swelling Effects 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- USFPINLPPFWTJW-UHFFFAOYSA-N tetraphenylphosphonium Chemical compound C1=CC=CC=C1[P+](C=1C=CC=CC=1)(C=1C=CC=CC=1)C1=CC=CC=C1 USFPINLPPFWTJW-UHFFFAOYSA-N 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- ITMCEJHCFYSIIV-UHFFFAOYSA-N triflic acid Chemical compound OS(=O)(=O)C(F)(F)F ITMCEJHCFYSIIV-UHFFFAOYSA-N 0.000 description 1
- JABYJIQOLGWMQW-UHFFFAOYSA-N undec-4-ene Chemical compound CCCCCCC=CCCC JABYJIQOLGWMQW-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H15/00—Compounds containing hydrocarbon or substituted hydrocarbon radicals directly attached to hetero atoms of saccharide radicals
- C07H15/18—Acyclic radicals, substituted by carbocyclic rings
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups
Abstract
The invention discloses an intermediate of a vaccine adjuvant MPLA, synthesis and application thereof. The intermediate provided by the invention takes the allyl phosphate ligand as a phosphate group source in the MPLA, and Nap is taken as a protecting group, so that the allyl phosphate ligand can be conveniently removed in the subsequent operation; the synthesized intermediate has short route and obviously increased total yield. Provides a basis for the synthesis and amplification of MPLA.
Description
Technical Field
The invention relates to an intermediate of a vaccine adjuvant MPLA, synthesis and application thereof.
Background
MPLA is the innermost liposomal (lipid a) fraction of endotoxin (LPS) from the cell wall of gram-negative bacteria, monophosphoryl lipid a (Monophosphoryl Lipid A, hereinafter MPL). Lipid A is an amphiphilic structure, also a critical structure for toxicity and immunogenicity of gram-negative bacteria, and can cause immune responses in the body. It can therefore be added as an adjuvant to a vaccine to increase the immunogenicity of the vaccine; generally, live attenuated vaccines, while more immunogenic, are also more virulent and potentially at risk of disease; and the non-toxic inactivated vaccine has weak immunogenicity. The addition of the adjuvant can enhance the immunogenicity of the inactivated vaccine and can not bring pathogenic risk to people with weak constitution. Simultaneously, the dosage of the antigen is reduced, and the vaccine supply quantity is increased. MPL is the first FDA-passing novel immunoadjuvant substance for humans other than aluminum salts. MPL acts on toll-like receptor 4 (TLR 4), and has the advantages of strong immunogenicity, clear mechanism, and low toxicity. The existing aluminum salt adjuvant mechanism is not clear, red swelling and pain occur at the injection site, the probability of the pain is large compared with MPL, and the aluminum salt adjuvant cannot generate specific cellular immunity. A number of new vaccine varieties using MPL adjuvant systems have been marketed both at home and abroad, such as the hepatitis b vaccine Fendrix, the cervical cancer vaccine Cerivix, the herpes zoster vaccine shinrix and the malaria vaccine mosquix. And twenty or more are in clinical stages.
Due to the structural differences in MPL derived from different bacteria or different serotypes of the same bacteria, the fundamental differences are the number/attachment positions of the fatty chains and the fatty chain carbon chain length, for example, the structures synthesized in this patent are shown below;
n1 and n3 are independently 10, 12 or 14, and n2 and n4 are independently 8, 10 or 12.
The existing MPL source depends on biological fermentation extraction, the cost is high, the problem of heterogeneity of lipopolysaccharide in the extraction process is difficult to solve, the purity problem is easy to be caused, and the potential safety hazard is brought. On the basis of special physicochemical properties and complex structures, the chemical method for synthesizing MPL is extremely difficult, and the MPL is difficult to replace biological fermentation. The total synthesis of MPL and its Lipid a analogues has also been reported to be limited to the order of milligrams, and the ligands that normally provide phosphate groups are typically benzyl pyrophosphate (di-O-benzoyloxy (N, N-diisophenylpyramine) or O-xylylene N, N-diethylphosphoramidite (N, N-diethyl-1,5-dihydro-3H-2,4,3-benzodioxapin-3-amine) as the source of phosphate groups, using Bn as the permanent protecting group. These protecting groups must finally be removed using hydrogenation conditions. For example, when similar lipid A and its derivatives are prepared in the prior art, benzyl is used, pd/C is required to react under the condition of atmospheric pressure or hydrogen pressurization for 10-20 h, the impurities are more, and the yield is 45% -68%. And the purification mode is complicated, and there is a mention that the regenerated cellulose needs to be used for filtration and ultrasonic removal of the catalyst; and then DEAE-cellulose ion exchange resin is used for separation, and a plurality of mixed solvents with different components are used as mobile phase to flush out the product. And repeatedly separating and concentrating to obtain a final product. (see ref 1:Alla Zamyatina,Harald Sekljic,Helmut Brade.Synthesis and purity assessment of tetra-and pentaacyl lipid A of Chlamydiacontaining (R) -3-hydroxy icosanoic acid. Tetrahedron 60 (2004) 12113-12137.Ref 2:Kaustabh K.Maiti,Michael DeCastro,Abu-Baker M.Abdel-Aal El-Sayd. Chemical Synthesis and Proinflammatory Responses of Monophosphoryl Lipid A Adjuvant Candida. Eur. J. Org. Chem.2010, 80-91).
The synthetic routes reported in the total synthesis of MPL analogues are longer; except that the benzyl protecting group used is difficult to completely remove at the later deprotection; to avoid side reactions in the reaction, some temporary protecting groups are usually needed, so that a plurality of protecting groups are involved, and repeated protection and deprotection are needed; in addition, the four fatty chains are linked in different orders in the reported literature, or four fatty chains (0+4) are linked after glycosylation, or 3+1 method is adopted, so that the fatty chain is not strong in connection selectivity, or the steric hindrance is larger, and the selectivity is lower during glycosylation. ( [1] J.AM.CHEM.SOC.2007,129,5200-5216; [2] WO 2013072768; [3] the chemical record, vol6,333-343 (2006) )
Therefore, in the current literature report, the hydrogenation result is not ideal, the yield is low, the impurities are more, the reaction time is long (generally more than 20 hours), the purification condition is complicated, and the large-scale production is difficult to meet the commercial demand.
Therefore, a new preparation method with a short route and a high total yield is urgently needed at present, so that a large amount of MPLA can be synthesized.
Disclosure of Invention
The invention aims to overcome the defect of lack of the existing preparation method of vaccine adjuvant MPLA, and provides an intermediate, synthesis and application of the vaccine adjuvant MPLA. The intermediate synthesized by the invention has short route and obviously increased total yield. The key intermediate of the MPLA is adopted, and the MPLA can be obtained after deprotection, thus providing a foundation for the synthesis and amplification of the MPLA.
The invention solves the technical problems through the following technical proposal.
The invention provides a preparation method of a compound shown as a formula 6, which comprises the following steps of;
in an organic solvent, in the presence of a base, a compound represented by formula 5 is mixed with(2, 2-trifluoro-N-phenylacetyl imine chlorine) to carry out imidization reaction as shown below to obtain the compound shown in the formula 6; wherein n1 is 10, 12 or 14, and n2 is 8, 10 or 12;
in the present invention, a compound of the formulaIs conventional in the art to mean the alpha configuration, the beta configuration, or a mixture of the alpha and beta configurations.
The imidization reaction can be carried out under conventional conditions in the imidization reaction; the following are preferred in the present invention:
the organic solvent can be halogenated hydrocarbon solvent, and the halogenated hydrocarbon solvent can be dichloromethane and/or chloroform.
The amount of the organic solvent is not particularly limited so as not to affect the reaction; for example, the mass to volume ratio of the compound represented by formula 5 to the organic solvent may be 10g/L to 100g/L (e.g., 50g/L to 80g/L, and further e.g., 65 g/L).
The base may be an organic base and/or an inorganic base as is conventional in this type of reaction in the art, for example 1, 8-diazabicyclo [5.4.0 ] ]Undec-7-ene (DBU), K 2 CO 3 DIPEA, naH and Cs 2 CO 3 One or more of the following; also e.g. DBU.
The molar ratio of the base to the compound of formula 5 may be 1:1 to 3:1 (e.g., 1.8:1 to 2.2:1, and also e.g., 2:1).
The molar ratio to the compound of formula 5 may be 1:1 to 10:1 (e.g., 5:1 to 8:1, and also 6:1).
The imidization reaction may be at a temperature of 0℃to 50℃such as 10℃to 30 ℃.
The imidization reaction can be performed under the protection of inert gas, and the inert gas can be nitrogen or argon.
The progress of the imidization reaction may be monitored by means conventional in the art (e.g., TLC, HPLC, or LCMS), typically at the end of the reaction when the compound of formula 5 is absent or no longer reduced in content; the imidization reaction is preferably carried out for a period of 0.2 to 5 hours (for example, 0.3 to 1 hour).
The post-treatment of the imidization reaction may be conventional in the art, for example, comprising the steps of: after the imidization reaction is finished, concentrating, separating and purifying. The separation and purification are preferably column chromatography, and the eluent of the column chromatography separation can be a mixed solution of alkalized petroleum ether and ethyl acetate (for example, petroleum ether/ethyl acetate=15:1, volume ratio, containing 0.1% Et by volume percent) 3 N)。
In one embodiment, n1 is 10, 12 or 14 (e.g., 10 or 12) and n2 is 10.
The preparation method of the compound shown in the formula 6 can also comprise the following steps; in an organic solvent, in the presence of a TBS removing protective agent, carrying out TBS removing protective group reaction on a compound shown in a formula 4 to obtain a compound shown in a formula 5; n1 and n2 are as defined above;
the operation and reaction conditions of the TBS protecting group removal reaction can be conventional operation and reaction conditions in the TBS protecting group removal reaction in the field; the following are preferred in the present invention:
in the TBS protecting group removal reaction, the organic solvent may be a cyclic ether solvent, and the cyclic ether solvent may be Tetrahydrofuran (THF).
The amount of the organic solvent is not particularly limited so as not to affect the reaction; for example, the mass to volume ratio of the compound of formula 4 to the organic solvent may be 5g/L to 100g/L (e.g., 10g/L to 50g/L, and further, e.g., 33 g/L).
The TBS removing protective agent can be a hydrofluoric acid pyridine complex (HF/pyridine, also called Olah reagent, wherein the mass percentage concentration of HF is 65-70%); preferably a pyridine solution of pyridine complex hydrofluoric acid (e.g., a solution of pyridine complex hydrofluoric acid diluted 3-6.5 times pyridine).
The volume to mass ratio of the detritus protectant to the compound of formula 4 may be 1mL/g to 20mL/g (e.g., 3 mL/g to 10 mL/g).
The temperature of the TBS protecting group removal reaction may be-80℃to 50 ℃ (e.g., -40℃to 30 ℃).
In one embodiment, the reaction for removing the TBS protecting group may comprise the following steps: the TBS removing protective agent is added into a mixed system of the compound shown in the formula 4 and the solvent to perform the TBS removing protective group reaction. The addition temperature may be from-70℃to-30 ℃ (e.g., -40 ℃). After the addition, the temperature of the reaction for removing the TBS protecting group can be 0-30 ℃.
The progress of the TBS removal reaction can be monitored by means conventional in the art (e.g., TLC, HPLC or LCMS), typically at the end of the reaction when the compound of formula 4 is absent or no longer reduced in amount; the reaction time for removing the TBS protecting group is preferably 1 to 24 hours (e.g., 2 to 12 hours).
The post-treatment of the TBS protecting group removal reaction may be conventional in the art, for example, comprising the steps of: and after the TBS protecting group removal reaction is finished, quenching, extracting with an organic solvent, drying, filtering, concentrating, separating and purifying. The quenching can be performed by using saturated sodium bicarbonate solution. The extracted organic solvent may be a halogenated hydrocarbon solvent (e.g., CH 2 Cl 2 ). The separation and purification are preferably column chromatography separation, and the eluent for the column chromatography separation can be petroleum ether and ethyl acetate (for example, petroleum ether: ethyl acetate=4:1, volume ratio).
The preparation method of the compound shown in the formula 6 can further comprise the following steps:
step (1), in an organic solvent, carrying out phosphorylation reaction on a compound shown as a formula 3 and an allyl ligand in the presence of tetrazole to obtain a mixture 1; the allyl ligand is hexadiene-N, N-diisopropyl phosphoramiditeCAS number 126429-21-8);
step (2), carrying out oxidation reaction on the mixture 1 and an oxidant to obtain the compound shown in the formula 3; n1 and n2 are as defined above;
the operation and reaction conditions of the phosphorylation reaction and the oxidation reaction may be those conventional in the art; the following are preferred in the present invention:
in step (1), the organic solvent may be a nitrile solvent (e.g., acetonitrile).
The amount of the organic solvent is not particularly limited so as not to affect the reaction; for example, the mass to volume ratio of the compound of formula 3 to the organic solvent may be 5g/L to 200g/L (e.g., 40g/L to 60g/L, and further e.g., 50 g/L).
In step (1), the molar ratio of tetrazole to the compound of formula 3 may be 1:1 to 10:1 (e.g., 3:1 to 5:1).
In step (1), the molar ratio of the allyl ligand to the compound of formula 3 may be from 1:1 to 5:1 (e.g., from 1:1 to 3:1, and further e.g., from 1.5:1 to 2:1).
In step (1), the temperature of the phosphorylation reaction may be-10℃to 50 ℃ (e.g., 10℃to 30 ℃).
In step (1), the progress of the phosphorylation reaction can be monitored by means conventional in the art (e.g., TLC, HPLC or LCMS), typically at the end of the reaction when the compound of formula 3 is absent or no longer reduced in amount; the time of the phosphorylation reaction is preferably 0.1 to 4 hours (e.g., 0.5 to 2 hours).
In step (2), the oxidizing agent may be m-chloroperoxybenzoic acid (mCPBA). Preferably, the oxidizing agent may be in the form of a solution of the halogenated hydrocarbon solvent (e.g., the mass to volume ratio of the oxidizing agent to the halogenated hydrocarbon solvent may be 0.01g/L to 0.05 g/L). The halogenated hydrocarbon solvent may be methylene chloride.
In step (2), the molar ratio of the oxidizing agent to the compound of formula 3 may be 1:1 to 5:1 (e.g., 2:1 to 3:1, and further e.g., 2.5:1 to 3:1).
In step (2), the temperature of the oxidation reaction may be-80℃to 10 ℃ (e.g. -40℃to-10 ℃).
In step (2), the progress of the oxidation reaction may be monitored by means conventional in the art (e.g., TLC, HPLC or LCMS), and the time of the oxidation reaction is preferably 0.1 to 4 hours (e.g., 0.5 to 2 hours).
In step (2), the post-treatment of the oxidation reaction may be conventional in the art, for example, comprising the steps of: after the oxidation reaction is finished, quenching, washing, drying, filtering, concentrating, separating and purifying. The quenching can be performed by using saturated sodium thiosulfate aqueous solution. The washing can be performed by using saturated sodium bicarbonate solution. The separation and purification are preferably column chromatography separation, and the packing for the column chromatography separation can be silica gel. The eluent for the column chromatography separation can be petroleum ether and ethyl acetate (the volume ratio of petroleum ether to ethyl acetate=8:1).
The preparation method of the compound shown in the formula 6 can further comprise the following steps: in an organic solvent, in a Borane (BH) 3 ) Lewis acid and H 2 In the presence of O, carrying out selective reduction ring-opening reaction on the compound shown in the formula 2 to obtain the compound shown in the formula 3; n1 and n2 are as defined above;
The operation and reaction conditions of the selective reduction ring-opening reaction can be conventional operation and reaction conditions in the selective reduction ring-opening reaction in the field; the following are preferred in the present invention:
in the selective reduction ring-opening reaction, the organic solvent may be a cyclic ether solvent, and the cyclic ether solvent may be Tetrahydrofuran (THF). The amount of the organic solvent is not particularly limited so as not to affect the reaction; for example, the mass to volume ratio of the compound of formula 2 to the organic solvent may be 10g/L to 200g/L (e.g., 50g/L to 150g/L, and further e.g., 100 g/L).
The borane may be in the form of a complex, e.g., BH 3 ·Me 3 N and/or BH 3 ·THF。
The molar ratio of the borane to the compound of formula 2 may be 1:1 to 5:1 (e.g., 3.5:1 to 4.5:1, and further e.g., 3.9:1).
The Lewis acid may be AlCl 3 。
The molar ratio of the borane to the lewis acid may be 1:1 to 1:3 (e.g., 1:1 to 1:2, and also e.g., 1:1.5).
Said H 2 The molar ratio of O to the compound of formula 2 may be 1:1 to 5:1 (e.g., 1.5:1 to 2.5:1, and also e.g., 1.9:1).
The temperature of the selective reduction ring-opening reaction may be-10 ℃ -50 ℃ (e.g., 10 ℃ -30 ℃).
The progress of the selective reduction ring-opening reaction can be monitored by means conventional in the art (e.g., TLC, HPLC or LCMS), typically at the end of the reaction when the compound of formula 2 is absent or no longer reduced in amount; the time for the selective reduction ring-opening reaction is preferably 0.1 to 4 hours (e.g., 0.5 to 2 hours).
The post-treatment of the selective reduction ring-opening reaction may be conventional in the art, for example, comprising the steps of: after the selective reduction ring-opening reaction is finished, quenching, extracting, drying, filtering, concentrating, separating and purifying. The quenching may be performed by adding water and hydrochloric acid solution (e.g., 1M HCl). The extraction solvent may be a halogenated hydrocarbon solvent (e.g., methylene chloride). The washing can be performed by using saturated sodium bicarbonate solution. The separation and purification are preferably column chromatography separation, and the packing for the column chromatography separation can be silica gel. The eluent for the column chromatography separation can be petroleum ether and ethyl acetate (volume ratio of petroleum ether to ethyl acetate=10:1).
The preparation method of the compound shown in the formula 6 can further comprise the following steps: in an organic solvent, in the presence of a condensing agent and a catalyst, carrying out esterification reaction of a compound shown in a formula 1 and a compound shown in a formula 19B to obtain the compound shown in a formula 2; n1 and n2 are as defined above;
The operation and reaction conditions of the esterification reaction can be conventional operation and reaction conditions in the esterification reaction of the type in the field; the following are preferred in the present invention:
in the esterification reaction, the organic solvent may be a halogenated hydrocarbon solvent, and the halogenated hydrocarbon solvent may be dichloromethane and/or chloroform. The amount of the organic solvent is not particularly limited so as not to affect the reaction; for example, the mass to volume ratio of the compound of formula 1 to the organic solvent may be 5g/L to 200g/L (e.g., 60g/L to 150g/L, and further e.g., 100 g/L).
The catalyst may be a catalyst conventional in this type of reaction in the art, for example an organic base, preferably one or more of 4-Dimethylaminopyridine (DMAP), triethylamine and pyridine, preferably DMAP.
The condensing agent may be one or more of 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride (edc·hcl), dicyclohexylcarbodiimide (DCC), and N, N' -Diisopropylcarbodiimide (DIC), for example 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride.
The molar ratio of the condensing agent to the compound of formula 1 may be 1:1 to 5:1 (e.g., 1:1 to 3:1, and further e.g., 1.2:1 to 2:1).
The molar ratio of the catalyst to the condensing agent may be from 0.01:1 to 1:1 (e.g., from 0.01:1 to 0.5:1, and further e.g., 0.05:1).
The molar ratio of the compound of formula 19B to the compound of formula 1 may be 1:1 to 3:1 (e.g., 1:1 to 2.5:1, and further e.g., 1.2:1).
The temperature of the esterification reaction may be-10 ℃ -50 ℃ (e.g., 10 ℃ -30 ℃).
The progress of the esterification reaction can be monitored by means conventional in the art (e.g., TLC, HPLC or LCMS), typically at the end of the reaction when the compound of formula 1 is absent or no longer reduced in amount; the time of the esterification reaction is preferably 1 to 24 hours (e.g., 2 to 12 hours).
The post-treatment of the esterification reaction may be conventional in the art, for example comprising the steps of: after the esterification reaction is finished, washing, drying, filtering, concentrating, separating and purifying. The washing can be sequentially carried out by using halogenated hydrocarbon solvents (such as dichloromethane) and saturated sodium bicarbonate solution. The separation and purification are preferably column chromatography separation, the packing of the column chromatography separation can be silica gel, and the eluent of the column chromatography separation can be petroleum ether and ethyl acetate (the volume ratio of petroleum ether to ethyl acetate=10:1).
The invention also provides a compound shown as the formula 6 and the formula 5;
wherein n1 and n2 are as defined above.
In one embodiment, the compound shown in formula 6 is any one of the following compounds:
in one embodiment, the compound shown in formula 5 is any one of the following compounds:
the invention provides a preparation method of a compound shown as a formula 5, which comprises the following steps: in an organic solvent, in the presence of a TBS removing protective agent, carrying out TBS removing protective group reaction on a compound shown in a formula 4 to obtain a compound shown in a formula 5; n1 and n2 are as defined above;
the reaction conditions and the operation in the preparation method of the compound shown in the formula 5 are as described above.
In the invention, if no special description exists, the room temperature is 10-30 ℃; "h" means hours; "overnight reaction" means a reaction for 8-16 hours.
The above preferred conditions can be arbitrarily combined on the basis of not deviating from the common knowledge in the art, and thus, each preferred embodiment of the present invention can be obtained.
The reagents and materials used in the present invention are commercially available.
The invention has the positive progress effects that: according to the invention, the allyl phosphate ligand is used as a phosphate group source in the MPLA, and Nap is used as a protecting group, so that the allyl phosphate ligand can be conveniently removed in the subsequent operation; the synthesized intermediate has short route and obviously increased total yield. Provides a basis for the synthesis and amplification of MPLA.
Detailed Description
The invention is further illustrated by means of the following examples, which are not intended to limit the scope of the invention. The experimental methods, in which specific conditions are not noted in the following examples, were selected according to conventional methods and conditions, or according to the commercial specifications.
Compound 18-a series examples
Preparation of starting Compound 1
To a reaction flask containing 2-deoxy-1-oxo- (1, 1-dimethylethyl) dimethylsilyl-2- [ (2, 2-trichloroethoxy) carbonyl ] amino-3, 4, 6-triacetyl-. Beta. -D-glucose (10 g,16.8 mmol) was slowly added guanidium hydrochloride buffer (100 mL, pH=8), stirred at room temperature for 3.5 hours, after the TLC detection of the consumption of the starting material, the reaction solution was neutralized with a cationic resin, filtered and concentrated, the product was extracted with methylene chloride and saturated sodium bicarbonate solution, and the organic layer was collected and concentrated to give 2-deoxy-1-oxo- (1, 1-dimethylethyl) dimethylsilyl-2- [ (2, 2-trichloroethoxy) carbonyl ] amino-. Beta. -D-glucose (1-1, 8.23 g).
1-1 and 2- (dimethoxymethyl) -naphthalene (5.1 g,25mmol,1.5 eq) were dissolved in 50mL of acetonitrile in a reaction flask, camphorsulfonic acid (0.39 g,1.69mmol,0.1 eq) was added, stirred at room temperature for 4h to react, triethylamine was added to neutrality, the reaction solution was extracted with dichloromethane and saturated sodium bicarbonate solution, and the solution was separated. The organic phase was dried and spun-dried to give a yellow solid. The crude product was passed through a silica gel sand funnel (PE: ea=5:1) to give product 1 (pale yellow solid, 6.97 g) in 68.3% yield in two steps.
Compound 1: 1 H NMR(400MHz,CDCl 3 )δ7.96–7.50(m,7H),5.72(s,1H),5.17(d,J＝6.3Hz,1H), 4.88(d,J＝7.7Hz,1H),4.73(q,J＝12.0Hz,2H),4.36(dd,J＝10.5,5.0Hz,1H),4.13–4.01(m,1H),3.86(t,J＝10.3Hz,1H),3.70–3.58(m,1H),3.52(td,J＝9.7,5.0Hz,1H),3.47–3.35(m,1H),2.96(s,1H),0.94 (d,J＝8.2Hz,9H),0.20–0.08(m,6H).
13 C NMR(101MHz,CDCl 3 )δ154.54,134.52,133.78,132.87,128.41,128.29,127.75,126.69,126.41, 126.07,123.93,101.97,96.33,95.30,81.52,74.85,70.71,68.68,66.20,60.73,26.94,25.59,17.90,-4.14,-5.26.
preparation of starting Compound 23
Compound 23 was prepared by reference to the procedure in the following references and ee values were determined using the same procedure: belma Hasdemir Hu lya Onar,/>Asymmetric synthesis of long chain beta-hydroxy fatty acid methyl esters as new elastase inhibitors tetrahedron: asymmetry (23) 2012,1100-1105. The title of the document is not standard and has been modified
Step (1) preparation of Compound 22
Mild acid (64.8 g,0.45 mol) and pyridine (48 mL) were dissolved in CH 2 Cl 2 To (100 mL) was added lauroyl chloride (21, 65.6g,0.3 mol) at 0deg.C. Stirred at room temperature for 2.5 hours. After complete consumption of the starting material, it was washed with 1M HCl (100 mL) and water (100 mL). The organic layer was dried, filtered and concentrated. It was dissolved in methanol (250 mL) and refluxed overnight. Concentrating the intermediate after consumption and passing through Al 2 O 3 Purification by column chromatography (toluene: ethyl acetate=2:1) afforded 22 (59.3 g, 77%) as a pale yellow solid.
1 H NMR(400MHz,CDCl 3 )δ3.70(3H,d,J＝1.5),3.42(2H,s),2.50(2H,t,J＝7.4),1.28(18H,s), 0.90(3H,t,J＝6.6).
13 C NMR(101MHz,CDCl 3 )δ202.51,167.58,137.67,128.96,128.15,125.25,52.02,48.84,42.90, 31.92,29.62,29.47,29.38,29.35,29.01,23.43,22.68,21.31,14.04.
Step (2) preparation of Compound 23
Preparation of ruthenium catalyst: 180mg (R) -Ru (OAc) 2 (BINAP) (diacetate [ (R) - (+) -2,2 '-bis (diphenylphosphino) -1,1' -binaphthyl)]Ruthenium (II), cas no: 325146-81-4) dissolved in CH 2 Cl 2 (5 mL) was added with 1.42N HCl (0.35 mL), and the mixture was stirred at room temperature for 1 hour and then dried by spin.
Compound 22 (15 g) and the prepared ruthenium catalyst were dissolved in methanol (50 mL). H 2 (1.5 MPa) stirring at 65℃for 6h. After completion of the starting material, the reaction solution was concentrated and purified by silica gel column chromatography (petroleum ether/ethyl acetate=5:1) to give compound 23 (white solid, 15g,98%, ee=98.7%).
1 H NMR(400MHz,CDCl 3 )δ4.01(1H,dq,J＝11.8,4.0),3.71(3H,s),2.90(1H,d,J＝4.0),2.46(2 H,ddd,J＝25.4,16.4,6.1),1.26(18H,s),0.88(3H,t,J＝6.8).
13 C NMR(101MHz,CDCl 3 )δ173.41,67.99,51.63,41.18,36.57,31.89,29.60,29.56,29.51,29.32, 25.47,22.65,14.05.
Reaction condition screening was performed on the preparation of compound 23:
relation of substrate to catalyst ratio (S/C) to ee value: the reaction is completed at 50 ℃ for about 4-6 hours with the S/C=300 catalyst amount, and the yield reaches more than 95 percent. To reduce the use of catalyst, a reaction of 180mg of catalyst per 20g of substrate (S/C > 300) was attempted, with a yield of only 71%.
Compound 22 (20 g) and 180mg of ruthenium catalyst prepared were dissolved in methanol (50 mL). H 2 Stirring at 65℃for 6h under (1.5 MPa). After completion of the starting material, the reaction solution was concentrated and purified by silica gel column chromatography (petroleum ether/ethyl acetate=5:1) to give compound 3 (white solid, 10.7g,71%, e)e value = 92.3%).
The ee value of the product under the different conditions is analyzed, the catalyst is less, the yield is reduced, and the ee value is only 92 percent. And several batches of S/c=300, all with ee values above 98%.
Example 1 preparation of Compound 24
Compound 23 (10 g,39mmol,1 eq) and 2-naphthaldehyde (18.14 g,116mmol,3 eq) were dissolved in THF (100 mL) and TMSOTF (6.88 g,31mmol,0.8 eq), (TMS) was added under ice-bath 2 O (37.68 g,232mmol,6 eq) and Et 3 SiH (15.7 g,135mmol,3.5eq). The reaction was carried out at 0℃for 1.5 hours, and the reaction mixture was treated with CH 2 Cl 2 (150 mL) and diluted with saturated NaHCO 3 And (5) washing. The organic layer was spin-dried and recrystallized (methylnaphthalene was obtained as a white solid at room temperature) (petroleum ether: ethyl acetate=5:1), filtered to remove impurities (methylnaphthalene). The filtrate (containing compound 24) was collected and dried to give crude product 24 as a pale yellow oily liquid which was used directly in the next step without purification. 1 H NMR(400MHz,CDCl 3 )0.89(3H,t，J6.7),1.27–1.64(18H,m),2.35(2H,dd,J 8.4)2.49 (1H,dd,J15.0,5.3),2.62(1H,dd,J15.0,7.3),3.68(3H,s),3.88(1H,m),4.54(2H,m),7.25–7.35(7H,m).
EXAMPLE 2 preparation of Compound 20
The compound 24 prepared in example 1 was dissolved in THF-H 2 O solution (5:1, 100 mL) was added to the solution, and the mixture was refluxed for 12h, followed by aqueous lithium hydroxide (9.41 g,224mmol,94 mL). After disappearance of starting material, it was cooled to room temperature and quenched by addition of 1.5M HCl in water to a pH of 7. The mixture was treated with CH 2 Cl 2 (150 mL) dilution with saturated NaHCO 3 And (5) washing. The organic phase was dried and spun-dried. Purification by column chromatography on silica gel (petroleum ether/ethyl acetate=5:1) afforded compound 20 (11.6 g, 77.9% yield in two steps, colorless sugar)Pulp).
1 H NMR(400MHz,CDCl 3 )δ10.00(1H,s,OH),7.85(4H,dd,J＝14.5,10.8),7.53(3H,d,J＝4.2), 4.85–4.71(2H,m,NapCH 2 O),4.08–3.94(1H,m,H-3),2.76(1H,dd,J＝15.2,6.9,H-2),2.64(1H,dd,J ＝15.2,4.3,H-2),1.84–1.58(2H,m,H-4),1.57–1.31(18H,m),1.00(3H,t,J＝6.0).
13 C NMR(101MHz,CDCl 3 )δ177.86,135.86,133.38,133.10,128.20,128.00,127.76,126.57,126.10, 125.99,125.89,75.94,71.71,39.81,34.37,32.05,29.77,29.71,29.48,25.27,22.82,14.26.
Example 3 preparation of Compound 2
Compound 19-A (5 g,11.7 mmol) was dissolved in CH with EDC. HCl (2.25 g,11.7mmol,1.2 eq) 2 Cl 2 (50 mL), stirring at room temperature for 15min, then adding compound 1 (5.93 g,9.8 mmol) and DMAP (0.06 g,0.5mmol,0.05 eq), stirring at room temperature for 10 h, and reacting the reaction mixture with CH in sequence 2 Cl 2 (100 mL) and saturated NaHCO 3 (60 mL) washing. The fractions were dried and concentrated and purified by silica gel column chromatography (petroleum ether/ethyl acetate=10:1) to give compound 2-a (8.6 g,86.7%, colorless syrup). TOF-MS: m/z 1036.46[ M+Na ]] + 。
δ H (400MHz,CDCl 3 )7.91(1H,s),7.82(3H,dt,J 9.3,4.9),7.54(1H,d,J 8.5),7.50–7.44(2H,m), 5.65(1H,s,NapCH),5.40–5.29(2H,m,H-3,NH),5.17(1H,dd,J 12.2,6.2,lipid-H-3),4.88(1H,d,J 7.8,H-1),4.69(2H,dd,J 48.4,12.0,Troc),4.34(1H,dd,J 10.5,4.9,H-6),3.84(1H,t,J 10.3,H-6),3.75(1H,t, J 9.4,H-4),3.62(1H,dd,J 16.8,7.7,H-2),3.54(1H,td,J 9.7,5.0,H-5),2.57(2H,ddd,J 20.5,15.2,6.3),2.12(2H,t,J 7.4),1.56–1.41(4H,m),1.32–1.13(34H,m),0.92–0.85(15H,m),0.11(6H,d,J 8.6).
δ C (101MHz,CDCl 3 )173.41,170.19,154.12,134.29,133.71,132.85,128.38,128.11,127.67,126.47, 126.16,125.81,123.73,101.76,96.82(C-1),95.31,78.95(C-4),74.66,71.14(C-3),69.98,68.72(C-6),66.47(C-5),59.11(C-2),39.26,34.31,33.83,31.93,29.63,29.50,29.35,29.29,29.06,25.53,25.07,24.93, 22.69,17.88,14.13,-4.21,-5.29.
EXAMPLE 4 preparation of Compound 3-a
Compound 2-a (2 g,1.97 mmol) was dissolved in THF (20 mL) and BH was added in sequence under ice-bath conditions 3 ·Me 3 N(0.57g,7.69 mmol，3.9eq)，AlCl 3 (1.52g,11.4mmol)，H 2 O (69 mg,3.83mmol,1.9 eq) was stirred at room temperature for 1.5h. Quenched with water (20 mL), 1M HCl solution (20 mL), and the reaction mixture was quenched with CH 2 Cl 2 (30 mL) was separated, dried and concentrated, and then purified by silica gel column chromatography (petroleum ether/ethyl acetate=10:1) to give compound 3-a (1.72 g,86%, colorless oily liquid). TOF-MS: m/z 1038.52[ M+Na ]] + .
δ H (400MHz,CDCl 3 )7.85–7.75(4H,m),7.46(3H,dd,J 7.7,4.0),5.37(1H,d,J 9.2,NH),5.15(1H, s,lipid-H-3),5.03(1H,t,J 9.7,H-3),4.81–4.71(4H,m,H-1,NapCH 2 ,Troc),4.63(1H,d,J 11.9,Troc), 3.81(2H,qd,J 10.5,3.8,H-6),3.69(1H,t,J 9.8,H-4),3.65–3.54(2H,m,H-2,H-5),3.47(1H,s,OH),2.62–2.48(2H,m,lipid-H-2),2.28(2H,t,J 7.3,lipid’-H-2),1.57(4H,s,lipid’-H-3,lipid-H-4),1.25(34H, s),0.88(15H,s),0.12(6H,d,J 17.5).
δ C (101MHz,CDCl 3 )174.35,171.77,154.27,135.54,133.29,133.02,128.20,127.89,127.71,126.29, 126.11,125.88,125.56,96.50(C-1),75.91(C-3),74.72(C-5),74.61,73.73,70.96,70.19(C-4),70.01(C-6),60.43,57.85(C-2),40.10,34.58,34.50,31.94,29.65,29.56,29.52,29.37,29.30,29.14,25.61,25.16,24.99, 22.71,21.06,17.93,14.21,14.14,-4.06,-5.28.
EXAMPLE 5 preparation of Compound 4
(1)
Compound 3-a (0.5 g,0.489 mmol) and tetrazole (102 mg,1.45mmol,3 eq) were dissolved in ultra-dry acetonitrile (10 mL), allylic ligand (hexadiene-N, N-diisopropylphosphoramidite, 0.24g,0.978mmol,2 eq) was added and after 40min reaction at room temperature the starting material was consumed.
The mCPBA (211 mg,1.22mmol,2.5 eq) was dissolved in ultra-dry dichloromethane (15 mL) at-40℃and the reaction was slowly warmed to-10℃and quenched with saturated sodium thiosulfate solution (20 mL) after 40min, washed with saturated sodium bicarbonate (40 mL), and purified by dry spin-dry silica gel column chromatography (petroleum ether: ethyl acetate=8:1) to give compound 4-a (0.53 g, 92%). TOF-MS: m/z 1198.67[ M+Na ]] + .
δ H (400MHz,CDCl 3 )7.85–7.77(4H,m),7.50–7.44(3H,m),5.90–5.73(2H,m,CH 2 ＝CH-CH 2 O-), 5.41–5.16(6H,m,NH,lipid-H-3,CH 2 ＝CH-CH 2 O-),4.97(1H,d,J 7.9,H-1),4.79-4.63(4H,td,J 12.3,12.4,12.0,Troc,Nap-CH 2 ),4.49–4.38(5H,m,H-4,CH 2 ＝CH-CH 2 O-),3.85(1H,d,J 9.6,H-6),3.75(1 H,d,J 5.4,H-6),3.72–3.65(1H,m,H-5),3.50–3.40(1H,m,H-2),2.64–2.60(2H,m),2.28(2H,t,J7.4),1.58(4H,br,J 7.3),1.25(34H,s),0.91–0.86(15H,m),0.13(6H,d,J 19.2).
δ C (101MHz,CDCl 3 )173.57,170.37,153.95,135.65,133.27,132.98,132.27,132.21,132.14,128.09, 127.87,127.68,126.23,126.05,125.81,125.65,118.56,118.40,95.63(C-1),95.32,74.59,74.12(C-5),74.06(C-4),73.61,72.42(C-3),70.06,68.70(C-6),68.62,68.56,68.46,68.41,58.67(C-2),39.59,34.51,34.24, 31.92,29.66,29.64,29.57,29.52,29.35,29.20,25.61,25.17,25.07,22.69,22.57,17.93,14.12,-4.15,-5.25.
(2)
Compound 3-a (0.5 g,0.489 mmol) and tetrazole (102 mg,1.45mmol,3 eq) were dissolved in ultra-dry acetonitrile (10 mL), allyl ligand (0.18 g,0.73mmol,1.5 eq) was added, after 2h reaction at room temperature the starting material was not consumed, 0.5eq of allyl ligand was added, and after 30min the starting material was consumed. mCPBA (211 mg,1.22mmol,2.5 eq) was dissolved in ultra-dry dichloromethane (15 mL) at-40 ℃, the reaction was added slowly to-10 ℃, quenched by addition of saturated sodium thiosulfate solution (20 mL) after 30min of reaction, washed with saturated sodium bicarbonate (40 mL), and purified by dry spin-dry silica gel column chromatography (petroleum ether: ethyl acetate=8:1) to give compound 4-a (0.49 g, 84.7%).
(3)
Compound 3-a (0.5 g,0.489 mmol) and tetrazole (171 mg,2.45mmol,5 eq) were dissolved in ultra-dry acetonitrile (10 mL), allyl ligand (0.24 g,0.978mmol,2 eq) was added, after 40min reaction at room temperature, the starting material was consumed, mCPBA (211 mg,1.22mmol,2.5eq) was dissolved in ultra-dry dichloromethane (15 mL) at-40 ℃, the reaction system was added, slowly warmed to-10 ℃, saturated sodium thiosulfate solution (20 mL) was added after 40min reaction quenching, saturated sodium bicarbonate washing (40 mL), and dry spin-dry silica gel column chromatography purification (petroleum ether: ethyl acetate=8:1) gave compound 4-a (0.474 g, 82.3%).
EXAMPLE 6 preparation of Compound 5-a
Compound 4-a (0.8 g,0.68 mmol) was dissolved in THF (24 mL), -HF/pyridine (2.4 mL, 65-70%) diluted in 15mL pyridine was added at 40 ℃. Slowly heating to room temperature for reaction for 12h, and using NaHCO to react the reaction solution 3 (40 mL) quenching, adding CH 2 Cl 2 (50 mL) was separated and purified by dry spin-dry silica gel column chromatography (petroleum ether: ethyl acetate=4:1) to give compound 5-a (0.657 g,91%, white solid). TOF-MS: m/z 1084.48[ M+Na ]] + 。
δ H (400MHz,CDCl 3 )7.85–7.77(4H,m),7.51–7.44(3H,m),5.91–5.65(2H,m,CH 2 ＝CH-CH 2 O-), 5.61(1H,d,J 9.4,NH),5.42–5.26(3H,m,H-1,CH 2 ＝CH-CH 2 O-),5.23–5.09(4H,m,H-3,CH 2 ＝CH- CH 2 O-,lipid-H-3),4.80-4.69(4H,m,Troc,Nap-CH 2 ),4.45–4.40(3H,m,H-4,CH 2 ＝CH-CH 2 O-),4.36– 4.27(2H,m,CH 2 ＝CH-CH 2 O-),4.25–4.19(1H,m,H-5),4.01–3.94(1H,m,H-2),3.84–3.75(2H,m,H- 6),2.60(2H,ddd,J 21.4,16.2,6.3),2.24(2H,t,J 7.6),1.56(4H,d,J 7.0),1.25(34H,s),0.88(6H,t,J 6.8).
δ C (101MHz,CDCl 3 )173.38,170.76,154.31,135.19,133.23,133.04,132.28,132.22,132.09,132.02, 128.23,127.92,127.70,126.67,126.15,125.95,125.81,118.66,118.38,95.37,91.50(C-1),74.65,73.63(C-4),70.82(C-3),69.90,69.57,69.52(C-5),68.67,68.61,68.47,68.40,68.35(C-6),54.27(C-2),39.10,34.46, 34.15,31.92,29.64,29.57,29.53,29.36,29.32,29.18,25.18,25.01,22.69,14.12.
EXAMPLE 7 preparation of Compound 6-a
Compound 5-a (1.3 g,1.22 mmol) was dissolved in ultra-dry (dry) CH 2 Cl 2 (20mL)，N 2 2, 2-trifluoro-N-phenylacetylimine chloride (1.52 g,7.32mmol,6 eq) was added under protection with DBU (371 mg,2.44mmol,2 eq) and reacted for 30mins at room temperature. The reaction solution was purified by column chromatography (petroleum ether: ethyl acetate=15:1 with 0.1% et) 3 N) to give compound 6-a (1.11 g,74% as colorless syrup). TOF-MS: m/z 1255.48[ M+Na ]] + 。
Example 8 preparation of Compound 7
(1)
Compound 1 (3.39 g,5.58 mmol) and compound 20 (4.29 g,11.16mmol,2 eq) were added to a reaction flask, 40mL of methylene chloride was added, EDC. HCl (2.14 g,11.16mmol,2 eq) and DMAP (135 mg,1.1mmol,0.2 eq) were added under ice bath, and after stirring at room temperature for 3h, compound 1 was consumed. The separated liquid was washed with saturated sodium bicarbonate solution (25 mL), dried, filtered, and purified by recrystallization (dichloromethane: methanol=1:12) to give compound 7 (5.08 g, 93.5%). TOF-MS: m/z 994.41[ M+Na ] ] + 。
δ H (400MHz,CDCl 3 )7.81(1H,s),7.76–7.61(7H,m),7.48–7.35(5H,m),7.30(1H,d,J 8.4),5.50 (1H,s,NapCH),5.39(1H,t,J 9.9,H-3),5.24(1H,d,J 9.1,NH),4.84(1H,d,J 7.8,H-1),4.67–4.62(3H,m,NapCH 2, -OCH 2 CCl 3 ),4.52(1H,d,J 11.8,-OCH 2 CCl 3 ),4.31(1H,dd,J 10.4,4.7,H-6),3.85–3.81(1H, m,Lipid-H-3),3.78(1H,d,J 10.3,H-6),3.71(1H,t,J 9.5,H-4),3.67–3.60(1H,m,H-2),3.53(1H,td,J9.5,5.1,H-5),2.74(1H,dd,J 14.9,6.0,Lipid-H-2),2.55(1H,dd,J 14.8,5.6,Lipid-H-2),1.54(2H,m, Lipid-H-4),1.27(18H,s),0.91(12H,s),0.13(6H,d,J 12.0).
δ C (101MHz,CDCl 3 )171.82,154.36,136.11,134.41,133.85,133.41,133.13,132.99,128.53,128.28, 128.23,128.11,127.88,127.85,126.60,126.51,126.31,126.20,126.04,125.96,125.91,123.85,101.88,97.14(C-1),79.18(C-4),75.73(C-6),74.88,71.39(C-3),71.28,68.91(C-6),66.76(C-5),60.64,59.29(C-2),39.77, 34.71,32.15,29.87,29.86,29.80,29.77,29.58,25.74,25.45,22.92,21.28,18.08,14.43,14.37,-3.98,-5.08.
(2)
EDC & HCl (1.087 g,5.67mmol,1.2 eq), compound 20 (2.18 g,5.67mmol,1.2 eq) was added to the reaction flask, 25mL of dichloromethane was added, after stirring at room temperature for 15min, compound 1 (2.87 g,4.72 mmol), DMAP (29 mg,0.237mmol, 0.05 eq) was added and the reaction was not completed for 12 h. The separated liquid was washed with saturated sodium bicarbonate solution (25 mL), dried, filtered and purified by column chromatography (petroleum ether: ethyl acetate=7:1) to give compound 7 (2.72 g, 59.23%).
(3)
EDC & HCl (1.81 g,9.44mmol,2 eq), compound 20 (2.54 g,6.6mmol,1.4 eq) was added to the reaction flask, 25mL of dichloromethane was added, after stirring at room temperature for 15min, compound 1 (2.87 g,4.72 mmol), DMAP (29 mg,0.237mmol, 0.05 eq) was added and the reaction was left unconsumed for 12 h. The separated liquid was washed with saturated sodium bicarbonate solution (25 mL), dried, filtered and purified by column chromatography (petroleum ether: ethyl acetate=7:1) to give compound 7 (3.27 g, 71.2%).
(4)
EDC & HCl (2.65 g,13.8mmol,2 eq), compound 20 (5.33 g,13.8mmol,2 eq) was added to the reaction flask, 50mL dichloromethane was added, compound 1 (4.12 g,6.78 mmol) was added after stirring at room temperature for 15min, DMAP (170mg,1.39mmol,0.2 eq) was consumed for 12 h. The separated liquid was washed with saturated sodium bicarbonate solution (50 mL), dried, filtered and purified by column chromatography (petroleum ether: ethyl acetate=7:1) to give compound 7 (5.22 g, 79.1%).
(5)
Compound 1 (3.39 g,5.58 mmol) and compound 20 (4.29 g,11.16mmol,2 eq) were added to a reaction flask, 40mL of methylene chloride was added, EDC. HCl (2.14 g,11.16mmol,2 eq) and DMAP (135 mg,1.1mmol,0.2 eq) were added under ice bath, and after stirring at room temperature for 3h, compound 1 was consumed. The separated liquid was washed with saturated sodium bicarbonate solution (25 mL), dried, filtered and purified by column chromatography (petroleum ether: ethyl acetate=7:1) to give compound 7 (4.71 g, 86.7%).
(6)
Compound 1 (3.39 g,5.58 mmol) and compound 20 (4.29 g,11.16mmol,2 eq) were added to a reaction flask, 40mL of methylene chloride was added, EDC. HCl (2.14 g,11.16mmol,2 eq) and DMAP (135 mg,1.1mmol,0.2 eq) were added under ice bath, and after stirring at room temperature for 3h, compound 1 was consumed. The separated liquid was washed with saturated sodium bicarbonate solution (25 mL), dried, filtered, and purified by recrystallization (dichloromethane: methanol=1:8) to give compound 7 (3.93 g, 72.4%).
Example 9 preparation of Compound 8
Compound 7 (6 g,6.16 mmol) was dissolved in dichloromethane (60 mL), acetic acid (12 mL, 2V) and zinc powder (6 g, 1V) (V means mass equivalent) were added, and the reaction was stirred vigorously at room temperature for 2 hours, followed by washing with saturated sodium bicarbonate (60 mL) and saturated brine (60 mL). The separated solution was dried and concentrated to obtain compound 8 (5.3 g). TOF-MS: m/z 821.134[ M+Na ] ] + . Directly used in the next reaction.
Example 10 preparation of Compound 9
(1)
The compound 8 prepared in example 9 was dissolved in methylene chloride (100 mL),FmocCl (3.2g,12.36 mmol,2eq), DIPEA (1.6 g,12.36mmol,2 eq) was added in ice bath, stirred at room temperature for 2h, washed with saturated brine (100 mL), dried in separate liquid and recrystallized (dichloromethane: methanol=1:10) to give compound 9 (5.6 g, 89.1%). TOF-MS: m/z 1042.7[ M+Na ]] + 。
δ H (400MHz,CDCl 3 )7.83(1H,s),7.78–7.65(7H,m),7.64–7.44(5H,m),7.40(6H,ddd,J 15.3, 9.7,5.3),7.32–7.24(3H,m),5.53(1H,s,NapCH),5.44(1H,t,J 9.4,H-3),4.97(1H,d,J 8.7,NH),4.91(1H,d,J 7.2,H-1),4.63(1H,d,J 11.8,Fmoc-CH 2 ),4.51(1H,d,J 11.8,,Fmoc-CH 2 ),4.32(1H,br,H-6), 4.28(2H,d,J 6.5,NapCH 2 ),4.17(1H,d,J 6.4,,Fmoc-CH),3.82(2H,s,H-6,Lipid-H-3),3.78–3.71(1H, m,H-4),3.67(1H,d,J 8.9,H-2),3.58(1H,br,H-5),2.70(1H,dd,J 14.8,6.3),2.50(1H,dd,J 14.8,5.5),1.55–1.37(2H,m),1.29–0.96(18H,m),0.91–0.81(12H,m),0.09(6H,d,J 13.5).
δ C (101MHz,CDCl 3 )155.78,143.79,141.23,135.92,134.24,133.63,133.17,132.87,132.78,128.31, 128.05,127.92,127.86,127.63,127.04,126.34,126.26,126.06,125.93,125.81,125.69,125.15,123.64,119.96,101.65,97.14(H-1),79.00(H-4),75.62(H-6),71.25(H-3),71.19,68.76(H-6),67.19,66.60(H-5), 58.98(H-2),47.04,39.74,34.53,31.92,29.61,29.53,29.34,25.49,25.16,22.69,17.86,14.14,-4.21,-5.36.
(2)
Compound 7 (6 g,6.16 mmol) was dissolved in dichloromethane (60 mL), acetic acid (12 mL, 2V) and zinc powder (6 g, 1V) were added, and the reaction was completed by vigorously stirring at room temperature for 2 hours, followed by saturated sodium bicarbonate (60 mL) and saturated brine (60 mL). The separated solution was dried and concentrated to obtain compound 8 (5.3 g). This was dissolved in methylene chloride (100 mL), fmocCl (3.2 g,12.36mmol,2 eq) was added under ice-bath, DIPEA (1.6 g,12.36mmol,2 eq) was stirred at room temperature for 2h, washed with saturated brine (100 mL), dried over liquid fraction, and recrystallized (methanol, 5V) to give compound 9 (4.91 g, 78.1%).
Example 11 preparation of Compound 10
Compound 9 (1.5 g,1.47 mmol) was reacted with molecular sieve1.5 g) dissolved in ultra-dry CH 2 Cl 2 (60mL)，N 2 Triethylsilane (0.55 mL,3.67mmol,2.5 eq) and PhBCl were added at-78deg.C under protection 2 (0.76 mL,5.88 mmol) and the reaction was stirred at-78deg.C for 1h. After the reaction, methanol (6 mL) is added for quenching, and triethylamine is added for regulating pH to 8. The reaction solution was filtered and saturated with NaHCO 3 (10 mL) washing. Purification by dry spin-dry silica gel column chromatography (petroleum ether: ethyl acetate=6:1) afforded compound 10 (1.2 g,80.1%, colorless oily liquid). TOF-MS: m/z 1044.45[ M+Na ]] + 。
δ H (400MHz,CDCl 3 )7.75–7.73(1H,m),7.70(7H,dd,J 14.7,5.4),7.66(2H,d,J 5.3),7.62(2H,d, J 5.8),7.53(1H,d,J 3.9),7.44–7.38(6H,m),7.30–7.25(3H,m),5.32(1H,t,J 9.8,H-3),4.93(1H,d,J9.3,NH),4.79(1H,d,J 6.6,H-1),4.73(2H,q,J 11.7,NapCH 2 ),4.57(2H,dd,J 24.5,11.7,Fmoc-CH 2 ), 4.24(2H,d,J 7.1,Lipid-NapCH 2 ),4.17–4.10(1H,m,Fmoc-CH),3.85-3.80(2H,s,H-6,Lipid-H-3),3.74 (1H,d,J 3.9,H-4),3.73–3.69(1H,m,H-6),3.60(1H,dd,J 18.4,9.4,H-2),3.49(1H,d,J 8.3,H-5),2.55(1H,dd,J 15.6,7.1),2.39(1H,dd,J 15.5,4.9),1.55–1.39(2H,m),1.22–1.06(18H,m),0.87–0.82(12 H,m),0.08(6H,d,J 13.4).
δ C (101MHz,CDCl 3 )172.00,155.84,143.86,141.23,135.99,135.05,133.23,133.15,132.98,132.92, 128.26,128.04,127.95,127.87,127.69,127.65,127.04,126.67,126.29,126.14,126.00,125.97,125.87,125.79,125.74,125.22,119.96,96.66(H-1),75.72(H-4),75.28(H-5),74.64(H-3),71.47(Fmoc-CH 2 ),67.24 (Lipid-NapCH 2 ),62.00(H-6),58.48(H-2),53.46,47.06(Fmoc-CH),39.74,34.19,31.96,29.67,29.65,29.62,29.60,29.38,25.52,25.15,22.73,17.90,14.17,-4.07,-5.25.
EXAMPLE 12 preparation of Compound 11
(1)
Into a reaction flask was charged acceptor compound 10 (606 mg,0.593 mmol), donor compound 6-a (1.1 g,0.89mmol,1.5 eq) molecular sieves600mg)，N 2 Adding super-dry CH under the condition 2 Cl 2 (10 mL) and TFOH (10. Mu.L, 0.12mmol,0.2 eq) diluted 100-fold with dichloromethane were added at-20deg.C and stirred for 40min at 20deg.C; methanol (5 mL) was added to quench, and triethylamine was adjusted to pH 8. CH for reaction solution 2 Cl 2 (35 mL) and saturated NaHCO 3 (10 mL) washing. The organic phase was concentrated by drying and purified by column chromatography (PE: ea=10:1) to give the glycosidation product 11-a (1.03 g, 84%). TOF-MS m/z 2088.14[ M+Na ]] + 。
δ H (400MHz,CDCl 3 )7.80–7.21(29H,m),5.89–5.68(2H,m,diallyl),5.48(1H,d,J 7.0,NH’),5.37 (1H,t,J 9.6,H-3’),5.32–5.10(6H,m,H-3,lipid-H-3,diallyl),4.87(1H,d,J 9.1,NH),4.80(1H,d,J 7.7,H-1’),4.74(1H,d,J 3.3,H-1),4.70(4H,d,J 12.8,Troc,Nap),4.64(2H,d,J 3.3,Nap-CH 2 ),4.53(2H,dd, J 29.1,11.9,Fmoc),4.45(1H,s,H-4’),4.43–4.31(5H,m,H-4,diallyl),4.24(2H,d,J 6.8,acceptor-lipid-Nap),4.16–4.11(1H,m,Fmoc-CH),4.07(1H,d,J 10.3,H-6),3.83(2H,d,J 10.2,H-6,acceptor-lipid-H- 3’),3.72(2H,dd,J 10.3,6.0,H-6’),3.65–3.54(3H,m,H-2,H-5,H-5’),3.41(1H,dd,J 17.7,8.4,H-2’),2.61(2H,t,J 6.6),2.50(1H,dd,J 15.6,7.1),2.35(1H,dd,J 15.8,5.1),2.28(2H,t,J 7.4),1.59(6H,d,J 7.7),1.42(4H,ddd,J 14.5,10.8,6.1),1.32–1.21(52H,m),0.86(18H,dd,J 14.9,7.8),0.15–0.05(6H, m).
δ C (101MHz,CDCl 3 )δ172.65,170.83,169.04,154.72,152.94,142.82,140.16,134.94,134.60,134.29, 132.19,132.15,132.08,131.91,131.84,131.82,131.29,131.22,131.13,131.06,129.85,127.80,127.12,127.06,126.93,126.85,126.79,126.62,126.60,126.55,125.97,125.32,125.17,125.10,125.02,124.90, 124.85,124.78,124.70,124.67,124.61,124.18,118.86,117.44,117.27,99.00(C-1’),95.53(C-1),94.38,75.08(C-5’),74.58(C-5)73.71(C-3),73.43(C-4’),73.31(C-4),73.11,73.05,72.96,72.91,72.60,71.37(C-3’), 70.37,69.10,67.55(C-6),67.49(C-6’),67.34,67.28,66.14,64.51,57.32(C-2),55.78(C-2’),46.00,38.78,38.64,33.50,33.41,33.13,30.89,28.67,28.63,28.61,28.59,28.54,28.51,28.31,28.17,24.54,24.15,24.07, 24.03,21.66,18.16,16.82,13.12,13.10,-4.91,-6.31.
(2)
Compound 6-a (50 mg,0.04 mmol) and compound 10 (51 mg,0.049mmol,1.2 eq) were placed in a reaction flask, 1mL of overdry dichloromethane was added under nitrogen, and TFOH (0.25 mg,0.00167mmol,0.05 eq) diluted 100-fold was added at-20 ℃. After 1h compound 6-a (donor) was consumed and compound 10 (acceptor) was not consumed. Adding methanol to quench triethylamine to be neutral. Purification by column chromatography (toluene: ethyl acetate=12:1) afforded compound 11-a (49 mg, 58.5%). Because product 11-a is nearly as polar as receptor 10, it is more polar than donor 6-a. Thus, when the acceptor 10 is excessive, although the reaction conversion is substantially equivalent, the isolation yield is low.
(3)
Compound 6-a (100 mg,0.081mmol,1.2 eq) and compound 10 (69 mg,0.067 mmol) were placed in a reaction flask, 1mL of overdry dichloromethane was added under nitrogen, and TFOH (0.5 mg,0.0033mmol,0.05 eq) diluted 100-fold was added at-20 ℃. After 1h, none of the donor acceptors was consumed, and the reaction time was unchanged. Adding methanol to quench triethylamine to be neutral. Purification by column chromatography (toluene: ethyl acetate=12:1) afforded compound 11-a (58 mg,0.0278mmol, 41.7%).
(4)
Compound 6-a (200 mg,0.162mmol,1.5 eq) and compound 10 (110 mg,0.107 mmol) were placed in a reaction flask, 1mL of ultra-dry dichloromethane was added under nitrogen, and 100-fold diluted TfOH (0.8 mg,0.0053mmol,0.05 eq) was added at-20 ℃. After 1h, none of the donor acceptors was consumed, and the reaction time was unchanged. Adding methanol to quench triethylamine to be neutral. Purification by column chromatography (toluene: ethyl acetate=12:1) afforded compound 11-a (107 mg, 48.2%).
(5)
Compound 6-a (200 mg,0.162mmol,1.5 eq) was reacted with compound10 (110 mg,0.107 mmol) was charged to a reaction flask, 1mL of ultra-dry dichloromethane was added under nitrogen, and TFOH (2.43 mg,0.0162mmol,0.15 eq) diluted 100-fold was added at-20 ℃. After 1h the donor was consumed. Adding methanol to quench triethylamine to be neutral. Purification by column chromatography (toluene: ethyl acetate=12:1) afforded compound 11-a (192 mg, 86.48%). TOF-MS: m/z 2088.14[ M+Na ] ] + 。
EXAMPLE 13 preparation of Compound 12-a
Compound 11-a (860 mg,0.029 mmol) was dissolved in CH in a reaction flask 2 Cl 2 (10mL)，N 2 Zinc powder (2.7 g, 40 mmol) and acetic acid (2.7 mL,45 mmol) were added and stirred at room temperature for 2h, after the reaction was completed, filtered, the filtrate azeotropically dried with toluene and column chromatographed (PE: EA=6:1) to give compound 12-a. TOF-MS m/z 1912.06[ M+Na ]] + 。
EXAMPLE 14 preparation of Compound 13-a (MPL-8)
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The compound 12-a prepared in example 13 was dissolved in ultra-dry DCM (10 mL), N 2 EDC & HCl (2 g,10.43 mmol) and fatty chain 19-A (1.48 g,3.47 mmol) were added at-10deg.C, and the reaction was stirred for 12h and purified by spin-dry column chromatography (toluene/ethyl acetate=10:1) to give compound 13-a (680 mg,71%, colorless clear syrup). TOF-MS: m/z 2323.15[ M+Na ]] + 。
EXAMPLE 15 preparation of Compound 14-a
Compound 13-a (580 mg,0.252 mmol) was dissolved in DMF (12 mL), N 2 Triethylamine (12 mL, 86 mmol) was added under stirring overnight at room temperature, and the reaction solution was dried by spin-drying and purified by column chromatography (PE: ea=6:1) to give compound 14-a. TOF-MS: m/z2030.2[M+Na] + 。
EXAMPLE 16 preparation of Compound 15-a
The compound 14-a prepared in example 15 was dissolved in ultra-dry CH 2 Cl 2 (5mL)，N 2 EDC & HCl (295 mg,1.53 mmol) and fatty chain 20 (298 mg,0.77 mmol) were added at room temperature, the reaction stirred at room temperature for 12h and purified by spin-dry column chromatography (toluene/ethyl acetate=5:1) to give compound 15-a (480 mg, yield 78% as colorless clear syrup). TOF-MS m/z 2468.4[ M+Na ] ] + 。
EXAMPLE 17 preparation of Compound 16-a
Compound 15-a (300 mg,0.123 mmol) was dissolved in THF (10 mL), -a solution of HF/Py (3 mL, 65-70%) in pyridine (9 mL) was added at 40 ℃. Warm to room temperature and stir overnight. After the reaction, saturated sodium bicarbonate aqueous solution is added for quenching, chloroform is added for extraction for a plurality of times. The organic layer was dried, filtered and concentrated, purified by C18 packing (CH 3 CN, meOH/ea=8:1, meOH eluting sequentially) gave 16-a (0.214 g,75%, white solid).
δ H (600MHz,CDCl 3 )7.77–7.24(28H,m),6.28(1H,d,J 9.5,NH),6.22(1H,d,J 7.4,NH’),5.77–5.70(1H,m,allyl),5.67–5.59(1H,m,allyl),5.44–5.31(3H,m,H-3,H-1’,H-3’),5.22–5.03(6H,m, allyl,H-1,lipid-H-3),4.99(1H,t,J 5.8,lipid-H-3),4.67–4.57(5H,m,Nap),4.52–4.42(3H,m,Nap),4.35–4.21(5H,m,H-4’,diallyl),4.17(1H,td,H-2),4.05(1H,d,J 7.2,H-5),3.87(1H,d,J 11.7,H-6’), 3.80–3.74(3H,m,lipid-H-3*2,H-6’),3.67–3.56(4H,m,H-4’,H-6*2,H-5’),3.32–3.28(1H,m,H-2’),3.27(1H,t,J 11.6,7.7,H-4),2.55–2.43(3H,m),2.28–2.11(9H,m),1.45–1.37(8H,m),1.22–1.10 (108H,m),0.80(18H,dt,J 7.0,5.1).
δ C (151MHz,CDCl 3 )173.28,172.55,170.87,170.29,169.85,168.98,135.06,134.48,133.92,132.31, 132.23,132.19,132.10,131.96,131.90,131.86,131.22,131.17,131.11,131.06,127.15,127.09,127.05,126.91,126.83,126.66,126.57,125.46,125.41,125.36,125.24,125.06,124.99,124.93,124.88,124.85, 124.82,124.73,124.64,124.61,117.45,117.30,98.02(C-1’),90.43(C-1),75.46(C-4),74.52,73.50,73.23,73.09,72.86(C-5’),72.51(C-3),71.64(C-3’),70.60(C-5),70.36,69.75,69.33,67.55(C-4’),67.51,67.28(C- 6’),67.24,66.43(C-6),59.37,55.37(C-2’),51.48(C-2),41.01,40.61,38.95,38.86,33.63,33.54,33.40,33.34,30.92,28.64,28.56,28.35,28.21,24.17,24.06,23.89,21.67,13.09.
EXAMPLE 18 preparation of Compound 17-a
(1)
Compound 16-a (240 mg,0.103 mmol), PPh 3 (25 mg,0.095 mmol) was added to the flask, after nitrogen protection, THF (10 mL), TEA (125. Mu.L, 0.9 mmol), HCOOH (75. Mu.L, 1.957 mmol), pd (Ph) 3 ) 4 (25 mg,0.02mmol, 0.2 eq). The reaction is carried out for 5 hours at 25 ℃ and is not completely reacted, the raw materials disappear after the continuous reaction is carried out for 12 hours, MS shows that the raw materials have undelivered all, and the raw materials are dried by spinning through a C18 column, namely CH 3 CN with 0.1％TEA,MeOH with 0.1％TEA,CH 2 Cl 2 MeOH with 0.1% TEA, in turn, gave compound 17-a (184 mg, 79.65%).
(2)
Compound 16-a (240 mg,0.103 mmol), PPh 3 (50 mg,0.190 mmol) was added to the flask, after nitrogen protection, THF (10 mL), TEA (250. Mu.L, 1.8 mmol), HCOOH (150. Mu.L, 3.914 mmol), pd (Ph) 3 ) 4 (50 mg,0.043mmol, 0.4 eq). After the reaction is carried out for 1.5 hours at 25 ℃, the reaction is complete, and the mixture is dried by spinning through a C18 column to obtain CH 3 CN with 0.1％TEA,MeOH with 0.1％TEA, CH 2 Cl 2 MeOH with 0.1% TEA gave compound 17-a (225 mg, 97.4%). TOF-M: M/z 2248.76[ M-H ]] + 。
Since there are two equivalents of all per molecule,when PPh is 3 And other reagents to conventional amounts (e.g., PPh 3 The molar ratio to all is about 0.9:1 to 1.1:1), compound 17 can be obtained in high yield.
EXAMPLE 19 preparation of Compound 18-a
(1)
Compound 17-a (50 mg,0.022 mmol) and DDQ (1.1 g,4.84mmol,220 eq) were added to a reaction flask and dried CHCl under nitrogen 3 (5 mL). After 60min of ultrasound at 30 ℃, the reaction was ended. One drop of triethylamine was added to quench the reaction, and the reaction was dried by spin-drying. Acetonitrile was added to the spin-dried reaction flask, and the mixture was poured into a column chromatography with a C18 packing to decolorize and purify. After complete removal of excess DDQ, CH 2 Cl 2 Meoh=3:1 gives the final product compound 18-a (25 mg, 67.6%). (since DDQ is greatly beyond the conventional dosage, in the case of small scale of charge, although the reaction yield can reach the theoretical value, a large amount of residual DDQ affects the yield of post-treatment purification)
(2)
Compound 17-a (50 mg,0.022 mmol) and DDQ (10 mg,0.044mmol,2 eq) were added to a reaction flask and dried CHCl under nitrogen 3 (5 mL). After 60min of ultrasound at 30 ℃, the reaction was ended. One drop of triethylamine was added to quench the reaction, and the reaction was dried by spin-drying. Acetonitrile was added to the spin-dried reaction flask, and the mixture was poured into a column chromatography with a C18 packing to decolorize and purify. After complete removal of excess DDQ, CH 2 Cl 2 Meoh=3:1 gives the final product compound 18-a (17 mg, 45.9%). If (4), the yield can be greatly improved by extending the reaction time.
(3)
Compound 17-a (50 mg,0.022 mmol) and DDQ (40 mg,0.177mmol,8 eq) were added to a reaction flask and dried CHCl under nitrogen 3 (5 mL). After 60min of ultrasonic treatment at 30 ℃, the reaction is complete. One drop of triethylamine was added to quench the reaction, and the reaction was dried by spin-drying. Acetonitrile was added to the spin-dried reaction flask, and the mixture was poured into a column chromatography with a C18 packing to decolorize and purify. After complete removal of excess DDQ, CH 2 Cl 2 :MeOH=3:1 gives the final product compound 18-a (29 mg, 78.4%). If (4), the yield can be greatly improved by extending the reaction time.
(4)
Compound 17-a (50 mg,0.022 mmol) and DDQ (40 mg,0.177mmol,8 eq) were added to a reaction flask and dried CHCl under nitrogen 3 (5 mL). After 80min of ultrasonic treatment at 30 ℃, the reaction is complete. One drop of triethylamine was added to quench the reaction, and the reaction was dried by spin-drying. Acetonitrile was added to the spin-dried reaction flask, and the mixture was poured into a column chromatography with a C18 packing to decolorize and purify. After complete removal of excess DDQ, CH 2 Cl 2 Meoh=3:1 to give the final product compound 18-a (35.8 mg,95.26%, HPLC purity: 98.5%). TOF-MS: m/z 1689.36[ M-H ]] + 。
Compound 18-b series examples
EXAMPLE 20 preparation of Compound 13-b
11-a (400 mg,0.194 mmol) was dissolved in CH in a reaction flask 2 Cl 2 (5mL)，N 2 Zinc powder 1.2g and acetic acid (1.2 mL) were added under the condition of stirring at room temperature for 2 hours, after the reaction was completed, the mixture was filtered, and the filtrate was azeotropically dried with toluene and subjected to column chromatography (PE: ea=6:1) to give compound 12-a (verified by MS to be TOF-MS: m/z 1915.49[ m+na)] + )
Compound 12-a was dissolved in ultra-dry DCM (5 mL), N 2 EDC & HCl (0.892 g, 4.650 mmol) and fatty chain 19-C (749 mg,1.552 mmol) (cas: 93390-37-5) were added at-10℃and the reaction stirred for 12h before purification by spin-dry column chromatography (toluene/ethyl acetate=10:1) gave compound 13-b (722.7 mg,63.4% as colorless, transparent syrup). TOF-MS m/z 2380.26[ M+Na ]] + 。
EXAMPLE 21 preparation of Compound 14-b
Compound 13-b (600 mg,0.254 mmol) was dissolved in DMF (12 mL), N in a reaction flask 2 Adding the triethylene under the conditionAmine (12 mL), stirred overnight at room temperature, and the reaction was purified by column chromatography (PE: ea=6:1) to give compound 14-b (verified by MS). TOF-MS m/z 2158.02[ M+Na ]] + 。
EXAMPLE 22 preparation of Compound 15-b
Dissolving Compound 14-b in ultra-Dry CH 2 Cl 2 (5mL)，N 2 EDC. HCl (390.35 mg,2.036 mmol) and fatty chain 20 (390.7 mg,1.016 mmol) were added at room temperature, the reaction stirred at room temperature for 12h and purified by spin-dry column chromatography (toluene/ethyl acetate=5:1) to give compound 15-b (504 mg,79.3% colorless clear syrup). TOF-MS m/z 2524.56[ M+Na ] + 。
EXAMPLE 23 preparation of Compound 16-b
Compound 15-b (500 mg,0.2 mmol) was dissolved in THF (18 mL), -a solution of HF/Py (5 mL, 65-70%) in pyridine (15 mL) was added at 40 ℃. Warm to room temperature and stir overnight. After the reaction, saturated sodium bicarbonate aqueous solution is added for quenching, chloroform is added for extraction for a plurality of times. The organic layer was dried, filtered and concentrated, purified by C18 packing (CH 3 CN, meOH/ea=8:1, meOH) to give 16-b (0.351 g,73.5%, white solid). TOF-MS m/z 2410.34[ M+Na ]] + 。
EXAMPLE 24 preparation of Compound 17-b
Compound 16-b (470 mg,0.196 mmol), PPh 3 (90 mg,0.370 mmol) was added to the flask, after nitrogen protection, THF (20 mL), TEA (490. Mu.L, 3.5 mmol), HCOOH (290. Mu.L, 7.8 mmol), pd (Ph) 3 ) 4 (90 mg,0.08 mmol). After the reaction is carried out for 1.5 hours at 25 ℃, the reaction is complete, and the mixture is dried by spinning through a C18 column to obtain CH 3 CN with 0.1％TEA,MeOH with 0.1％TEA,CH 2 Cl 2 MeOH with 0.1% TEA gave compound 17-b (428 mg, 94.3%). TOF-M M/z 2330.16[ M+Na ]] + 。
EXAMPLE 25 preparation of Compound 18-b
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Compound 17-b (90 mg,0.039 mmol) and DDQ (71 mg,0.312mmol,8 eq) were added to a reaction flask and dried CHCl under nitrogen 3 (10 mL). After 80min of ultrasonic treatment at 30 ℃, the reaction is complete. One drop of triethylamine was added to quench the reaction, and the reaction was dried by spin-drying. Acetonitrile was added to the spin-dried reaction flask, and the mixture was poured into a column chromatography with a C18 packing to decolorize and purify. After complete removal of excess DDQ, CH 2 Cl 2 Meoh=3:1 gives the final product compound 18-b (65 mg, 95.26%). TOF-MS: m/z 1745.47[ M-H ]] + . HPLC purity: 97.5%.
Compound 18-c series examples
EXAMPLE 26 preparation of Compounds 2-c
Compound 19-B (cas: 163310-36-9) (2.313 g,5.93 mmol) was dissolved in CH with EDC. HCl (1.136 g,5.93mmol,1.2 eq) 2 Cl 2 (25 mL), stirring at room temperature for 15min, then adding compound 1 (3 g,4.94 mmol) and DMAP (0.03 g,0.247mmol, 0.05 eq), stirring at room temperature for 10h, and reacting the reaction mixture with CH in turn 2 Cl 2 (50 mL) and saturated NaHCO 3 (30 mL) washing. The fractions were dried and concentrated and purified by silica gel column chromatography (petroleum ether/ethyl acetate=10:1) to give compound 2-c (4.29 g,84.3%, colorless syrup). TOF-MS: m/z 1052.7[ M+Na ]] + 。
EXAMPLE 27 preparation of Compound 3-c
The compound 2-c is treated1g,0.97 mmol) dissolved in THF (10 mL) and added BH in sequence under ice-bath conditions 3 ·Me 3 N(0.28g,3.783 mmol，3.9eq)，AlCl 3 (0.76g,5.7mmol)，H 2 O (35 mg,1.92mmol,1.9 eq) was stirred at room temperature for 1.5h. Quenched with water (10 mL), 1M HCl solution (10 mL), and the reaction mixture was quenched with CH 2 Cl 2 (15 mL) was separated, dried and concentrated, and then purified by silica gel column chromatography (petroleum ether/ethyl acetate=10:1) to give compound 3-c (0.882 g,87.6%, colorless oily liquid). TOF-MS: m/z 1059.42[ M+Na] + .
EXAMPLE 28 preparation of Compound 4-c
Compound 3-c (0.5 g, 0.480 mmol) and tetrazole (102 mg,1.45mmol,3 eq) were dissolved in ultra-dry acetonitrile (10 mL), allyl ligand (hexadiene-N, N-diisopropylphosphoramidite, 0.236g,0.964mmol,2 eq) was added and after 40min reaction at room temperature the starting material was consumed. mCPBA (207.9 mg,1.205mmol,2.5 eq) was dissolved in ultra-dry dichloromethane (15 mL) at-40 ℃, the reaction was added slowly to-10 ℃, quenched by the addition of saturated sodium thiosulfate solution (20 mL) after 40min, washed with saturated sodium bicarbonate (40 mL), and purified by dry spin-dry silica gel column chromatography (petroleum ether: ethyl acetate=8:1) to give compound 4-c (0.537 g, 93.3%). TOF-MS: m/z 1214.8[ M+Na ]] + .
EXAMPLE 29 preparation of Compound 5-c
Compound 4-c (1.5 g,1.26 mmol) was dissolved in THF (45 mL), -HF/pyridine (4.8 mL, 65-70%) diluted in 30mL pyridine was added at 40 ℃. Slowly heating to room temperature for reaction for 12h, and using NaHCO to react the reaction solution 3 (80 mL) quenching, adding CH 2 Cl 2 (100 mL) was separated and purified by dry spin-dry silica gel column chromatography (petroleum ether: ethyl acetate=4:1) to give compound 5-c (1.26 g,93.4%, white solid). TOF-MS: m/z 1100.53[ M+Na ]] + 。
EXAMPLE 30 preparation of Compound 6-c
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Compound 5-c (1.3 g,1.21 mmol) was dissolved in ultra-dry CH 2 Cl 2 (20mL)，N 2 2, 2-trifluoro-N-phenylacetylimine chloride (1.50 g,7.24mmol,6 eq) was added under protection with DBU (365 mg,2.42mmol,2 eq) and reacted for 30mins at room temperature. The reaction solution was purified by column chromatography (petroleum ether: ethyl acetate=15:1 with 0.1% et) 3 N) to give compound 6-c (1.19 g,79% as colorless syrup). TOF-MS: m/z 1271.65[ M+Na ]] + 。
Example 31 preparation of Compound 11-c
Into a reaction flask was charged acceptor compound 10 (700 mg,0.684 mmol), donor compound 6-c (1.28 g,1.026mmol,1.5 eq) molecular sieves600mg)，N 2 Adding super-dry CH under the condition 2 Cl 2 (10 mL) and TFOH (20. Mu.L, 0.137mmol,0.2 eq) diluted 100-fold with dichloromethane were added at-20deg.C and stirred for 40min at 20deg.C; methanol (5 mL) was added to quench, and triethylamine was adjusted to pH 8. CH for reaction solution 2 Cl 2 (35 mL) and saturated NaHCO 3 (10 mL) washing. The organic phase was concentrated by drying and purified by column chromatography (PE: ea=10:1) to give the glycosidation product 11-c (1.27 g, 89%). TOF-MS m/z 2104.9[ M+Na ]] + 。
EXAMPLE 32 preparation of Compound 12-c
11-c (500 mg,0.24 mmol) was dissolved in CH in a reaction flask 2 Cl 2 (5mL)，N 2 Zinc powder 1.5 g) and acetic acid (15 mL) were added thereto and stirred at room temperature for 2 hours, and after the completion of the reaction, the mixture was filteredAzeotropic spin-drying of the filtrate with toluene and column chromatography (PE: EA=6:1) gave compound 12-c (MS-verified TOF-MS: m/z 1929.5[ M+Na ]] + )。
EXAMPLE 33 preparation of Compound 13-c
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Compound 12-c was dissolved in ultra-dry DCM (5 mL), N 2 EDC & HCl (1.1 g,5,76 mmol) and fatty chain 19-A (819 mg,1.92 mmol) were added at-10deg.C and the reaction stirred for 12h before purification by spin-dry column chromatography (toluene/ethyl acetate=10:1) to give compound 13-c (447mg, 79.4% as colorless clear syrup). TOF-MS m/z 2338.23[ M+Na ] ] + 。
EXAMPLE 34 preparation of Compound 14-c
Compound 13-c (600 mg, 0.299 mmol) was dissolved in DMF (12 mL), N 2 Triethylamine (12 mL) was added under stirring overnight at room temperature, and the reaction solution was purified by column chromatography (PE: ea=6:1) to give compound 14-c (verified by MS). TOF-MS m/z 2115.9[ M+Na ]] + 。
Example 35 preparation of Compound 15-c
Dissolving Compound 14-c in ultra-Dry CH 2 Cl 2 (5mL)，N 2 EDC & HCl (397.2 mg,2.072 mmol) and fatty chain 20 (398.4 mg,1.036 mmol) were added at room temperature, the reaction stirred at room temperature for 12h and purified by spin-dry column chromatography (toluene/ethyl acetate=5:1) to give compound 15-c (518.8 mg,81.4% as colorless, transparent syrup). TOF-MS m/z 2482.47[ M+Na ]] + 。
EXAMPLE 36 preparation of Compound 16-c
Compound 15-c (400 mg,0.162 mmol) was dissolved in THF (18 mL), -a solution of HF/Py (4 mL, 65-70%) in pyridine (12 mL) was added at 40 ℃. Warm to room temperature and stir overnight. After the reaction, saturated sodium bicarbonate aqueous solution is added for quenching, chloroform is added for extraction for a plurality of times. The organic layer was dried, filtered and concentrated, purified by C18 packing (CH 3 CN, meOH/ea=8:1, meOH) to give 16-c (0.293 g,76.9%, white solid). TOF-MS m/z 2368.26[ M+Na ]] + 。
EXAMPLE 37 preparation of Compound 17-c
Compound 16-c (459 mg,0.196 mmol), PPh 3 (90 mg,0.370 mmol) was added to the flask, after nitrogen protection, THF (20 mL), TEA (490. Mu.L, 3.5 mmol), HCOOH (290. Mu.L, 7.8 mmol), pd (Ph) 3 ) 4 (90 mg,0.08 mmol). After the reaction is carried out for 1.5 hours at 25 ℃, the reaction is complete, and the mixture is dried by spinning through a C18 column to obtain CH 3 CN with 0.1％TEA,MeOH with 0.1％TEA,CH 2 Cl 2 MeOH with 0.1% TEA gave compound 17-c (405 mg, 91.5%). TOF-MS m/z 2288.05[ M+Na ]] + 。
EXAMPLE 38 preparation of Compound 18-c
Compound 17-c (100 mg,0.044 mmol) and DDQ (80 mg,0.353mmol,8 eq) were added to a reaction flask and dried CHCl under nitrogen 3 (10 mL). After 80min of ultrasonic treatment at 30 ℃, the reaction is complete. One drop of triethylamine was added to quench the reaction, and the reaction was dried by spin-drying. Acetonitrile was added to the spin-dried reaction flask, and the mixture was poured into a column chromatography with a C18 packing to decolorize and purify. After complete removal of excess DDQ, CH 2 Cl 2 Meoh=3:1 to give the final product compound 18-c (68 mg, 91.5%). TOF-MS: m/z 1703.4[ M-H ]] + . HPLC purity: 96.5%
Comparative example 1
Compound 17 (50 mg,0.022 mmol) and Pd (400 mg) were added to a hydrogenation kettle, dissolved in THF: h 2 O=4:1 (20 mL), 1mpa,30 ℃ for 24h. One drop of triethylamine was added to quench, and the reaction solution was filtered and dried. Decolorization and purification in column chromatography using C18 packing gave the final product compound 18 (12 mg, 32.4%). The TLC plate layer shows that the raw materials are remained, the impurity points are more, and the raw materials are not reduced after the prolonged time. The yields and purities are significantly worse than in the examples described above.
As can be seen from the above, (1) the present invention uses allyl ligands to avoid subsequent hydrogenation reactions, which can be completed with tetraphenylphosphine at 1.5 h. The final product can be obtained after simple decolorization by using a C18 packing column. The problems of more impurities, lower yield and complex purification mode caused by adopting hydrogenation to remove benzyl protecting groups in the prior art are avoided. Has obviously better effect.
(2) Compared with the prior art that palladium hydrocarbon reaction is needed for more than 20 hours, and then ion column chromatography is repeatedly filtered, the yield is lower (only about 50 percent) and the method using the Nap protecting group simplifies the operation, the yield of the deprotection step can reach more than 91.5 percent after optimization, and the purity reaches 97 percent (the determination method refers to the HPLC-ELSD method for determining the content of MPL in the BLP25 liposome vaccine, and the method comprises the following steps of 2012, 5 th, wang Mingjuan, wang and Hu Changqin).
Claims (15)
1. A method for preparing a compound represented by formula 6, comprising the steps of;
in an organic solvent, in the presence of a base, a compound represented by formula 5 is mixed withCarrying out imidization reaction as shown below to obtain the compound shown in the formula 6; wherein n1 is 10, 12 or 14, and n2 is 8, 10 or 12;
The organic solvent is halogenated hydrocarbon solvent;
the mass volume ratio of the compound shown in the formula 5 to the organic solvent is 10g/L-100g/L;
the alkali is organic alkali and/or inorganic alkali;
the molar ratio of the alkali to the compound shown in the formula 5 is 1:1-3:1;
the molar ratio of the compound shown in the formula 5 to the compound is 1:1-10:1;
the imidization reaction temperature is 0-50 ℃;
2. the process for producing a compound of formula 6 according to claim 1,
the imidization reaction is carried out under the protection of inert gas;
or, the post-treatment of the imidization reaction comprises the following steps: concentrating, separating and purifying after the imidization reaction is finished;
or n1 is 10, 12 or 14, and n2 is 10.
3. The method for producing a compound represented by formula 6 according to claim 2,
the organic solvent is dichloromethane and/or chloroform;
or the mass volume ratio of the compound shown in the formula 5 to the organic solvent is 50g/L-80g/L;
or, the base is 1, 8-diazabicyclo [5.4.0]Undec-7-ene, K 2 CO 3 DIPEA, naH and Cs 2 CO 3 One or more of the following;
or, the molar ratio of the alkali to the compound shown as the formula 5 is 1.8:1-2.2:1;
or alternatively, the first and second heat exchangers may be,the molar ratio of the compound shown in the formula 5 to the compound is 5:1-8:1;
or, the imidization reaction temperature is 10-30 ℃;
or, when the imidization reaction is performed under the protection of inert gas, the inert gas is nitrogen or argon;
or, in the post-treatment step of imidization, the separation and purification is column chromatography separation, and the eluent for the column chromatography separation is a mixed solution of alkalized petroleum ether and ethyl acetate;
or n1 is 10 or 12, and n2 is 10.
4. A process for producing a compound represented by the formula 6 according to claim 3,
in the eluent for column chromatographic separation, the volume ratio of petroleum ether to ethyl acetate is=15:1, and the eluent contains 0.1 percent of Et by volume percent 3 N。
5. The method for producing a compound represented by formula 6 according to any one of claims 1 to 4, further comprising the steps of; in an organic solvent, in the presence of a TBS removing protective agent, carrying out TBS removing protective group reaction on a compound shown in a formula 4 to obtain a compound shown in a formula 5; n1 and n2 are as defined in any one of claims 1 to 3;
The organic solvent is a cyclic ether solvent;
the mass volume ratio of the compound shown in the formula 4 to the organic solvent is 5g/L-100g/L;
the TBS removing protective agent is a hydrofluoric acid pyridine complex;
the volume-mass ratio of the TBS removal protective agent to the compound shown in the formula 4 is 1-20 mL/g;
the temperature of the reaction for removing the TBS protecting group is-80 ℃ to 50 ℃;
6. the method for producing a compound represented by formula 6 according to claim 5,
the TBS protecting group removal reaction comprises the following steps: adding the TBS removing protective agent into a mixed system of the compound shown in the formula 4 and the solvent to perform TBS removing protective group reaction; the addition temperature is between-70 ℃ and-30 ℃; after the addition, the temperature of the reaction for removing the TBS protecting group is 0-30 ℃;
or, the post-treatment of the TBS protecting group removal reaction comprises the following steps: and after the TBS protecting group removal reaction is finished, quenching, extracting with an organic solvent, drying, filtering, concentrating, separating and purifying.
7. The method for producing a compound represented by formula 6 according to claim 6,
In the post-treatment step of TBS protecting group removal reaction, saturated sodium bicarbonate solution is adopted for quenching;
or in the post-treatment step of TBS protecting group removal reaction, the extracted organic solvent is halogenated hydrocarbon solvent;
or, in the post-treatment step of TBS protecting group removal reaction, the separation and purification are column chromatography separation;
or, in the post-treatment step of TBS protecting group removal reaction, the eluent separated by column chromatography is petroleum ether and ethyl acetate.
8. The process for producing a compound of formula 6 according to claim 7,
in the post-treatment step of TBS protecting group removal reaction, the extracted organic solvent is CH 2 Cl 2 ；
Or, in the post-treatment step of TBS protecting group removal reaction, the eluent separated by column chromatography is petroleum ether and ethyl acetate in a volume ratio of=4:1.
9. The process for producing a compound of formula 6 according to claim 8,
in the TBS protecting group removal reaction, the organic solvent is tetrahydrofuran;
or the mass volume ratio of the compound shown in the formula 4 to the organic solvent is 10g/L-50g/L;
Or when the TBS removing protective agent is a pyridine hydrofluoric acid complex, the pyridine hydrofluoric acid complex is a pyridine solution;
or the volume-mass ratio of the TBS removal protective agent to the compound shown in the formula 4 is 3-10 mL/g;
or the temperature of the reaction for removing the TBS protecting group is-40 ℃ to 30 ℃;
or, the TBS protecting group removal reaction adopts the following steps: adding the TBS removing protective agent into a mixed system of the compound shown in the formula 4 and the solvent to perform TBS removing protective group reaction; the adding temperature is-40 ℃; after the addition, the temperature of the reaction for removing the TBS protecting group is 0-30 ℃;
or, in the post-treatment step of TBS protecting group removal reaction, saturated sodium bicarbonate solution is adopted for quenching; the extracted organic solvent is halogenated hydrocarbon solvent; the separation and purification is column chromatography separation, and the eluent for the column chromatography separation is petroleum ether and ethyl acetate.
10. The method for preparing a compound represented by formula 6 according to claim 5, further comprising the steps of:
step (1), in an organic solvent, carrying out phosphorylation reaction on a compound shown as a formula 3 and an allyl ligand in the presence of tetrazole to obtain a mixture 1; the allyl ligand is hexadiene-N, N-diisopropyl phosphoramidite;
Step (2), carrying out oxidation reaction on the mixture 1 and an oxidant to obtain the compound shown in the formula 3; n1 and n2 are as defined in claim 5;
11. the method for preparing a compound represented by formula 6 according to claim 10, further comprising the steps of:
in an organic solvent in borane, lewis acid and H 2 In the presence of O, carrying out selective reduction ring-opening reaction on the compound shown in the formula 2 to obtain the compound shown in the formula 3; n1 and n2 are as defined in claim 10;
12. the method for preparing a compound represented by formula 6 according to claim 11, further comprising the steps of:
in an organic solvent, in the presence of a condensing agent and a catalyst, carrying out esterification reaction of a compound shown in a formula 1 and a compound shown in a formula 19B to obtain the compound shown in a formula 2; n1 and n2 are as defined in claim 11;
13. a compound shown as a formula 6 and a formula 5;
wherein n1 and n2 are as defined in any one of claims 1 to 3.
14. A compound according to claim 13 of formula 6 or formula 5;
Wherein the compound shown in the formula 6 is any one of the following compounds:
wherein the compound shown in the formula 5 is any one of the following compounds:
15. a method for preparing a compound represented by formula 5, comprising the steps of:
in an organic solvent, in the presence of a TBS removing protective agent, carrying out TBS removing protective group reaction on a compound shown in a formula 4 to obtain a compound shown in a formula 5; n1 and n2 are as defined in claim 13 or 14;
the reaction conditions and operations in the preparation method of the compound shown in the formula 5 are as defined in any one of claims 5 to 10.
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| Chemical Synthesis of Burkholderia Lipid A Modified with Glycosyl Phosphodiester-Linked 4-Amino-4-deoxy-β-L-arabinose and Its Immunomodulatory Potential;Hollaus, Ralph等;《Chemistry - A European Journal》;第21卷(第10期);4102-4114 * |
| Dioxane-type (2-naphthyl)methylene acetals of glycosides and their hydrogenolytic transformation into 6-O- and 4-O-(2-naphthyl)methyl (NAP) ethers;Borbas, Aniko等;《Tetrahedron》;第58卷(第28期);5723-5732 * |
| Modulating Heparanase Activity: Tuning Sulfation Pattern and Glycosidic Linkage of Oligosaccharides;Zhu, Sanyong等;《Journal of Medicinal Chemistry》;第63卷(第8期);4227-4255 * |
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