Hydrochloric Acid → (-)-Asparagine

HRID 595610Notes

Reaction equation

HRID 595610 reaction equation
Cl.C1C2C(COC(NCCOCCOCCN)=O)C3C(=CC=CC=3)C=2C=CC=1.C[N+]1(C2N=C(OC)N=C(OC)N=2)CCOCC1.[Cl-].Cl.C[O:49][C:50](=[O:57])[C@H:51]([CH2:53][C:54](=[O:56])[NH2:55])[NH2:52].Cl>CS(C)=O>[NH2:52][C@H:51]([C:50]([OH:57])=[O:49])[CH2:53][C:54](=[O:56])[NH2:55]

Participants

6

Reactant 4

HCID 313Hydrochloric AcidHCID 27340594-(4,6-Dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride2×HCID 27562241-(9-Fluorenylmethyloxycarbonyl-amino)-3,6-dioxa-8-octaneamine hydrochlorideHCID 6453494Methyl L-asparaginate monohydrochloride

Product 1

HCID 6267(-)-Asparagine

Reagents, catalysts & solvents 1

HCID 679Dimethyl SulfoxideSolvent

Procedure

1-(9-fluorenylmethyloxycarbonyl-amino)-3,6-dioxa-8-octanamine hydrochloride (Fmoc-EDOBEA, Iris Biotech GmbH) was added to an anhydrous DMSO solution (5 mg/mL) of HA-TBA synthesized using HA-Na (99 kDa) as a starting material in Example 1-2 in an amount of 0.1 molar equivalents relative to the HA unit. Next, DMT-MM was added in an amount of 0.2 molar equivalents relative to the HA unit, and stirring was performed at room temperature for 6 hours. Then, L-asparagine methyl ester hydrochloride and DMT-MM were added in this order in amounts of 5 and 3-6 molar equivalents, respectively, relative to the HA unit, and stirring was performed at room temperature overnight. The reaction solution was dialyzed for purification against DMSO. Piperidine (Wako Pure Chemical Industries, Ltd.) was added to the resulting dialyzate to g a concentration of 20%, and stirring was performed for 2 hours to remove the Fmoc group. Thereafter, the mixture was dialyzed for purification against DMSO, an aqueous solution of 0.3 M NaCl, and distilled water in this order. 5N NaOH solution was added to the resulting dialyzate to adjust the pH to at least 12.5, and stirring was performed for an hour to perform ester hydrolysis and carboxy deprotection. Thereafter, the mixture was neutralized with 5N HCl solution and further dialyzed for purification against distilled water and ultrapure water in this order, and then the dialyzate was freeze-dried to give HA-Asn/EDOBEA.

Workup

  1. customsynthesized
  2. stirringrespectively, relative to the HA unit, and stirring
  3. waitwas performed at room temperature overnight
  4. customThe reaction solution was dialyzed for purification against DMSO
  5. additionPiperidine (Wako Pure Chemical Industries, Ltd.) was added to the resulting dialyzate
  6. stirringstirring
  7. waitwas performed for 2 hours
  8. customto remove the Fmoc group
  9. customThereafter, the mixture was dialyzed for purification against DMSO
  10. distillationan aqueous solution of 0.3 M NaCl, and distilled water in this order
  11. addition5N NaOH solution was added to the resulting dialyzate
  12. stirringstirring
  13. waitwas performed for an hour
  14. customester hydrolysis and carboxy deprotection
  15. customfurther dialyzed for purification
  16. distillationagainst distilled water and ultrapure water in this order
  17. customthe dialyzate was freeze-dried

Sources & evidence

ORD record
ord-0ac1f7b853fa40d3a6a7f2698377a5e4
Source dataset
uspto-grants-2016_01
Patent number
US09243077B2

Reaction notes

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