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CN-117362375-B - Intermediate for synthesizing 3D-MPL, preparation method and application thereof

CN117362375BCN 117362375 BCN117362375 BCN 117362375BCN-117362375-B

Abstract

The invention relates to the technical field of pharmaceutical chemistry, in particular to an intermediate for synthesizing 3D-MPL, a preparation method and application thereof. A first intermediate for the synthesis of 3D-MPL is selected from the group of compounds represented by the following structural formulae: Wherein n1 and n2 are respectively selected from any integer between 8 and 12. The protecting groups in the intermediate can be removed efficiently and conveniently, the route for synthesizing 3D-MPL is obviously shortened, and the total yield is obviously improved.

Inventors

  • GAO QI
  • CHEN DEXIANG
  • ZHU JIALE
  • ZHENG ZHIWEI
  • CHEN CHAO
  • ZHOU DANDAN

Assignees

  • 成都迈科康生物科技有限公司

Dates

Publication Date
20260508
Application Date
20231129

Claims (20)

  1. 1. The preparation method of the 3D-MPL is characterized by comprising the step of synthesizing by using a sixth intermediate shown in the following structural formula: wherein n1 to n6 are respectively selected from any integer between 8 and 12; the preparation method of the sixth intermediate comprises the following steps: In an organic solvent, carrying out a reduction reaction on the compound 1 under the action of acetic acid and zinc powder to obtain a compound 3, and carrying out a selective reduction ring-opening reaction on the compound 3 in the presence of triethylsilane and dichlorophenyl borane to obtain a compound 4; The compound 4 reacts with the compound A to form a first intermediate, wherein the first intermediate is a compound 5 shown in the following structural formula, and the structural formula of the compound A is shown as follows: ; In an organic solvent, carrying out glycosylation reaction on the first intermediate and a compound B in the presence of an acid catalyst to obtain a second intermediate, wherein the second intermediate is a compound 6 shown in the following structural formula, and the structural formula of the compound B is shown as follows: ; in an organic solvent, under the condition of zinc powder/acid catalysis, carrying out a Troc protecting group removal reaction at the 2-position of the second intermediate to obtain a third intermediate, wherein the third intermediate is a compound 7 shown in the following structural formula; In an organic solvent, carrying out amidation condensation reaction on the third intermediate and a compound C under the catalysis of a condensing agent to obtain a fourth intermediate, wherein the fourth intermediate is a compound 8 shown in the following structural formula, and the structural formula of the compound C is shown as follows: ; In an organic solvent, under the condition of desilication ether catalysis, carrying out a silyl ether protecting group removal reaction on the fourth intermediate to obtain a fifth intermediate, wherein the fifth intermediate is a compound 9 shown in the following structural formula; in an organic solvent, under the condition of palladium catalysis, carrying out allyloxy protecting group removal reaction on the fifth intermediate to obtain a sixth intermediate, wherein the sixth intermediate is a compound 10 shown in the following structural formula; Specifically, the sixth intermediate is synthesized with reference to the following synthesis route: ; the compound 10 forms 3D-MPL with reference to the following synthetic pathway: 。
  2. 2. The method of claim 1, wherein the first intermediate is selected from any one of the compounds represented by the following structural formulas: 。
  3. 3. The preparation method according to claim 1, wherein the conditions for synthesizing the compound 5 comprise a molar ratio of the compound A to the compound 4 of 1-3:1, an acylation reaction temperature of-10 to 50 ℃, a mass volume ratio of the compound 4 to the solvent of 50-200g/L using a halogenated hydrocarbon solvent, and a molar ratio of the condensing agent to the compound 4 of 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride, dicyclohexylcarbodiimide and N, N' -diisopropylcarbodiimide of 1-5:1; the progress of the acylation reaction was monitored, and the end point of the reaction was set at the point when the compound 4 disappeared or the content was no longer reduced, and the post-treatment was performed after the end of the acylation reaction.
  4. 4. The preparation method according to claim 1, wherein the conditions for synthesizing the compound 5 comprise a molar ratio of the compound A to the compound 4 of 1-2.5:1, an acylation reaction temperature of 10-30 ℃, a solvent of dichloromethane and/or chloroform, a condensing agent of 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride, a molar ratio of the condensing agent to the compound 4 of 1-3:1, and a reaction time of 2-12 hours; the post-treatment comprises washing, drying, filtering, concentrating and separating and purifying the target product.
  5. 5. The preparation method according to claim 1, wherein the reaction conditions for synthesizing the compound 4 comprise that the organic solvent is a halogenated hydrocarbon solvent, and the mass-volume ratio of the compound 3 to the organic solvent is 50-200g/L; the molar ratio of the triethylsilane to the compound 3 is 2-5:1; The molar ratio of the dichlorophenyl borane to the compound 3 is 2-5:1; the reaction temperature is-90 to 40 ℃; The progress of the reaction was monitored, and the end point of the reaction was set at the point when the compound 3 disappeared or the content was no longer reduced, and after the end of the reaction, post-treatment was performed.
  6. 6. The process according to claim 5, wherein the reaction conditions for synthesizing compound 4 comprise that the organic solvent is methylene chloride and/or chloroform; The molar ratio of triethylsilane to compound 3 is 3:1; the molar ratio of the dichlorophenyl borane to the compound 3 is 3:1; the reaction temperature is-78 ℃ and the reaction time is 0.5-2 hours; the post-treatment comprises quenching the reaction system after the reaction is finished, filtering, washing, drying, concentrating, separating and purifying.
  7. 7. The process according to claim 1, wherein the conditions for synthesizing compound 3 comprise that the organic solvent is a halogenated hydrocarbon solvent; the mass volume ratio of the compound 2 to the organic solvent is 50-200 g/L; The mass ratio of the acetic acid to the compound 2 is 2-5:1; the mass ratio of the zinc powder to the compound 2 is 1-3:1; the reaction temperature is 0-30 ℃; The progress of the reaction is monitored, the end point of the reaction is the point when the compound 2 disappears or the content is no longer reduced, and the post-treatment is carried out after the reaction is finished.
  8. 8. The process according to claim 7, wherein the conditions for synthesizing compound 3 comprise that the organic solvent is methylene chloride and/or chloroform; The mass-volume ratio of the compound 2 to the organic solvent is 100 g/L; the mass ratio of the acetic acid to the compound 2 is 3.5:1; the mass ratio of the zinc powder to the compound 2 is 2:1; The reaction temperature is 10-20 ℃, and the reaction time is 3-6 h; The post-treatment comprises the steps of filtering, washing, drying, concentrating, separating and purifying the reaction system in sequence after the reaction is finished.
  9. 9. The process according to claim 1, wherein the conditions for synthesizing compound 2 comprise that the organic solvent is a halogenated hydrocarbon solvent; The mass volume ratio of the compound 1 to the organic solvent is 50-200 g/L; the acid binding agent is any one of pyridine, triethylamine and tetramethyl ethylenediamine; the molar ratio of the acid binding agent to the compound 1 is 1-3:1; the molar ratio of the allyl chloroformate to the compound 1 is 2-5:1; the reaction temperature is 0-30 ℃; the progress of the reaction was monitored, and the end point of the reaction was the point at which the compound 1 disappeared or the content was no longer reduced, and after the end of the reaction, post-treatment was performed.
  10. 10. The process according to claim 9, wherein the conditions for synthesizing compound 2 comprise that the organic solvent is methylene chloride and/or chloroform; the mass volume ratio of the compound 1 to the organic solvent is 100 g/L; The acid binding agent is tetramethyl ethylenediamine; the molar ratio of the acid binding agent to the compound 1 is 1-2:1; the molar ratio of the allyl chloroformate to the compound 1 is 3-4:1; The reaction temperature is 10-20 ℃, and the reaction time is 1-5 h; The post-treatment comprises quenching, washing, drying, filtering, concentrating, separating and purifying the reaction system in sequence after the reaction is finished.
  11. 11. The method of claim 1, wherein the second intermediate is selected from any one of the compounds of the following structural formulas: 。
  12. 12. The method according to claim 1, wherein the conditions for synthesizing the second intermediate include that the organic solvent is a halogenated hydrocarbon solvent; the mass volume ratio of the first intermediate to the organic solvent is 50-200 g/L; the acid catalyst is one or more of trifluoromethanesulfonic acid, trifluoromethanesulfonic acid trimethylsilyl ester and trifluoromethanesulfonic anhydride; The molar ratio of the acid catalyst to the first intermediate is 3-5:1; the molar ratio of the compound B to the first intermediate is 5:8-10; the reaction temperature is-10 ℃ to-40 ℃; monitoring the progress of the glycosylation reaction, taking the first intermediate as a reaction end point when the first intermediate disappears or the content is not reduced any more, and carrying out post-treatment after the reaction is finished.
  13. 13. The method according to claim 12, wherein the conditions for synthesizing the second intermediate include that the organic solvent is methylene chloride and/or chloroform; the acid catalyst is trifluoromethanesulfonic acid; the molar ratio of the acid catalyst to the first intermediate is 4-5:1; the molar ratio of the compound B to the first intermediate is 5:8; the reaction temperature is-20 ℃, and the reaction time is 2-12 h; The post-treatment comprises quenching the reaction system after the reaction is finished, concentrating, separating and purifying.
  14. 14. The method of claim 1, wherein the third intermediate is selected from any one of the compounds of the following structural formulas: 。
  15. 15. The method according to claim 1, wherein the conditions for synthesizing the third intermediate include that the organic solvent is a halogenated hydrocarbon solvent; the mass volume ratio of the second intermediate to the organic solvent is 50-200 g/L; the acid is selected from one or more of acetic acid, hydrochloric acid and sulfuric acid; the mass volume ratio of the acid to the second intermediate is 1-5 g/mL; The molar ratio of the zinc powder to the second intermediate is 20-15:1; The reaction temperature is 10-30 ℃; The progress of the reaction is monitored, the end point of the reaction is taken as the point when the second intermediate disappears or the content is no longer reduced, and the post-treatment is carried out after the reaction is finished.
  16. 16. The method according to claim 15, wherein the conditions for synthesizing the third intermediate include that the organic solvent is methylene chloride and/or chloroform; the acid is acetic acid; the mass to volume ratio of the acid to the second intermediate is 3g/mL; the molar ratio of the zinc powder to the second intermediate is 16:1; The reaction temperature is room temperature, and the reaction time is 2-6h; The post-treatment comprises the steps of washing, separating and purifying the reaction system in sequence after the reaction is finished.
  17. 17. The method of claim 1, wherein the fourth intermediate is selected from any one of the compounds of the following structural formulas: 。
  18. 18. The method according to claim 1, wherein the conditions for synthesizing the fourth intermediate include that the organic solvent is a halogenated hydrocarbon solvent; the mass volume ratio of the third intermediate to the organic solvent is 120-200 g/L; The condensing agent is one or more of 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride, dicyclohexylcarbodiimide and N, N' -diisopropylcarbodiimide; the molar ratio of the condensing agent to the third intermediate is 5-10:1; The molar ratio of the compound C to the third intermediate is 5-10:1; The reaction temperature is 10-30 ℃; The progress of the reaction is monitored, the end point of the reaction is taken as the point when the third intermediate disappears or the content is no longer reduced, and the post-treatment is carried out after the reaction is finished.
  19. 19. The method according to claim 8, wherein the conditions for synthesizing the fourth intermediate include that the organic solvent is methylene chloride and/or chloroform; the condensing agent is 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride; The molar ratio of the condensing agent to the third intermediate is 6:1; the molar ratio of the compound C to the third intermediate is 6:1; The reaction temperature is room temperature, and the reaction time is 2-6h; the post-treatment comprises the steps of washing, spin drying, separating and purifying the reaction system in sequence after the reaction is finished.
  20. 20. The method of claim 1, wherein the fifth intermediate is selected from any one of the compounds of the following structural formulas: 。

Description

Intermediate for synthesizing 3D-MPL, preparation method and application thereof Technical Field The invention relates to the technical field of pharmaceutical chemistry, in particular to an intermediate for synthesizing 3D-MPL, a preparation method and application thereof. Background Immunoadjuvants are a class of agents that alter primarily cytokine levels to enhance vaccine efficacy, typically by activating MHC molecules, costimulatory factors, or intracellular-related signaling molecules, among other modes. A typical class of endotoxin Lipopolysaccharide (LPS) in the cell wall of gram-negative bacteria, the LPS-mediated immune activation mechanism is the production of inflammatory factors such as Th1 interferons, chemokines, etc. by LPS interacting with TLR4 (toll-like receptors) and the receptor protein MyD88, TRIF. The most highly evaluated family molecule is monophosphoryl lipid a (Monophosphoryl Lipid A, hereinafter MPL). Lipid A is an amphiphilic structure, is an attenuated LPS derivative, retains better immunogenicity and has basically the same action mechanism as LPS. The toxicity of MPL was significantly reduced, between 0.1% -1% compared to LPS, and proved to be significantly effective. Are therefore widely used as vaccines, allergy drugs and adjuvants for immunotherapy to enhance immune responses. Generally, live attenuated vaccines are more immunogenic but more virulent, whereas non-virulent inactivated vaccines are less immunogenic. The addition of the adjuvant can enhance the immunogenicity of the inactivated vaccine and does not bring about pathogenic risk. MPL is the first FDA-passing novel immunoadjuvant substance for humans other than aluminum salts. The natural extract of MPL mainly comprises Salmonella minnesota extract, 3D-MPL as main ingredient, and Escherichia coli extract EcML, MPL-12 as main ingredient. And a plurality of new vaccine varieties adopting an MPL adjuvant system are commercially available at home and abroad, such as hepatitis B vaccine Fendrix, cervical cancer vaccine Cerivix, herpes zoster vaccine Shingrix and malaria vaccine Mosquirix. Most of the prior MPL sources are biological extraction, and because the MPL from different bacteria or different serotypes of the same bacteria has different structures, the basic difference is the number/connection position of the fatty chains and the carbon chain length of the fatty chains, most of the MPL sources are mixtures, the mixing proportion is floating, the quality is unstable, and the MPL sources have great potential safety hazard when used as an adjuvant. Meanwhile, compared with the naturally extracted MPL, the synthetic MPLA has high cytokine induction level on immunized mice, CD4+ T cells and CD8+ T cells have equivalent induction level, but the synthetic MPLA can induce and activate NKT cells. Synthetic MPL is a current hot spot, and has various GLA (MPLA), 3D-MPL and the like which are evaluated clinically at present, but has a complex structure, a longer route and the following synthetic difficulties: 1. The ligands which typically provide phosphate groups are typically benzyl pyrophosphate (di-O-benzyloxy (N, N-diisopropylamino) phosphine) or O-xylylene N, N-diethylphosphoramidite as a source of phosphate groups, using Bn as a protecting group. These protecting groups must finally be removed using hydrogenation conditions, and purification is complex, making it difficult to achieve large-scale production. 2. The current synthetic route reported by the total synthesis of MPL analogues is long, the benzyl protecting group is difficult to completely remove in the later deprotection, and in order to avoid side reactions, a plurality of protecting groups are usually needed, so that the deprotection needs to be repeatedly protected, and the route is long, the selectivity is poor, the yield is low and the impurities are many. Aiming at the problems, the inventor has developed a highly effective synthetic process of the MPLA, and considers that the 3D-MPL is more stable than the MPLA in chemical structure, and preclinical experiments show that the anti-inflammatory, antiviral and the like are better than the MPLA, and the immune mouse antibody titer IgG can be obviously improved. We have further developed 3D-MPL that can be synthesized on a large scale. Based on this, the present invention has been proposed. The 3D-MPL structure is shown below; Wherein n1 to n6 are independently any integer between 8 and 12, preferably n1, n3 and n5 are independently 10,12 and 14, n2, n4 and n6 are independently 8, 10 or 12. Disclosure of Invention The invention aims to provide an intermediate for synthesizing 3D-MPL, a preparation method and application thereof. The embodiment of the invention provides a series of novel intermediates for synthesizing 3D-MPL, and the protecting groups in the intermediates can be removed efficiently and conveniently, so that the route for synthesizing 3D-MPL is obviously shortened, and the total yield is