Preparation method of bivalirudin

A bivalirudin and molar ratio technology, applied in the field of peptide synthesis, can solve the problems of many process control points, complex steps of bivalirudin, unstable process, etc.

Active Publication Date: 2013-08-14
QILU PHARMA HAINAN +1
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Problems solved by technology

[0007] Due to the complex steps of liquid-phase synthesis of bivalirudin, many process control points, many by-products, and unstable process, therefore, bivalirudin is usually prepared by solid-phase synthesis.
Chinese patent document CN101555274A adopts a step-by-step coupling strategy to prepare bivalirudin. When constructing the -Gly-Gly-Gly-Gly-structure in the molecule, Fmoc-Gly-OH is used for step-by-step coupling, which cannot avoid bivalirudin[- Production of Gly], [+Gly] impurities and bivalirudin [-2Gly], [+2Gly] impurities
The polarity of these impurities is very similar to that of bivalirudin itself, and it is difficult to completely remove them during the purification process, and the product yield cannot be effectively improved, resulting in a decrease in product...

Method used

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  • Preparation method of bivalirudin
  • Preparation method of bivalirudin
  • Preparation method of bivalirudin

Examples

Experimental program
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Effect test

Embodiment 1

[0061] Embodiment 1, the preparation of Fmoc-Pro-Gly-Gly-Gly-Gly-OH

[0062] Weigh tetrapolyglycine (24.6g, 100mmol) and sodium carbonate (21.2g, 200mmol) into 500ml water, stir for 10 minutes to dissolve, add Fmoc-Pro–OSu (47.7g, 110mmol) of 1,4-dioxane The solution was 500ml, stirred and reacted at room temperature for 2.5 hours, TLC detected the end of the reaction, concentrated under reduced pressure to remove 1,4-dioxane, adjusted the pH of the aqueous phase with 2N hydrochloric acid, and when the pH was 2-3, a large amount of solids were precipitated, filtered with suction, Washed with water, dried, and purified by column chromatography to obtain 51.2 g of the target product with a yield of 90.0% and a purity of 98.7%. MS m / z: 566 (M+1). [α] D 20℃ =-24.3°(DMF,c=1.0).

[0063] h 1 NMR (600MHz, CD 3 SOCD 3 ):

[0064] δ8.34(dd,J=4.8Hz,2H),8.24(dd,J=6.0Hz,1H),8.21(dd,J=6.0Hz,1H),8.11-8.16(m,3H),8.06( dd,J=5.4Hz,1H),7.90(q,4H),7.68(t,J=9.0Hz,2H),7.64(d,J=7.8Hz,1H),7...

Embodiment 2

[0066] Example 2, preparation of side chain fully protected bivalirudin peptide-resin

[0067] Weigh 25.0g of Fmoc-Leu-Wang resin (loading capacity 0.4mmol / g, 10mmol) into the solid phase reactor, add 150ml of 20% piperidine DMF solution, stir and react at 25-30°C for 10min, repeat deprotection 3 times , suction filtration after completion of the reaction, the resin was washed with 200ml DMF, suction filtration, repeated washing 6 times, Fmoc-Tyr ( t Bu)-OH (MW: 459,30mmol) 13.8g, 1-hydroxybenzotriazole (HOBt) (HOBt) (MW: 135.1,30mmol) 3.9g were dissolved in 100ml DMF, added to the solid phase reactor, and then N,N - Diisopropylcarbodiimide (DIC) (MW: 126.2, 30mmol) 3.6ml, react at 25-35°C for about 2 hours, the end point of the reaction is subject to the negative test of ninhydrin, repeat the above steps, follow the The rutin peptide sequences are coupled with the corresponding Fmoc protected amino acids one by one, and the molar equivalents of the protected amino acids and ...

Embodiment 3

[0069] Example 3, preparation of side chain fully protected bivalirudin peptide-resin

[0070] Weigh 20.0g of Fmoc-Leu-Wang resin (loading capacity 0.5mmol / g, 10mmol) into the solid phase reactor, add 150ml of 20% piperidine DMF solution, stir and react at 25-30°C for 10min, repeat deprotection 3 times , suction filtration after completion of the reaction, the resin was washed with 200ml DMF, suction filtration, repeated washing 6 times, Fmoc-Tyr ( t Bu)-OH (MW:459,30mmol) 13.8g, N-hydroxy-7-azabenzotriazole (HOAt) (MW:136.1,30mmol) 4.1g, 2-(7-aza-1H- Benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (HATU) (MW:380.2,30mmol) 11.4g was dissolved in DMF and added to the solid phase reactor, Add 5ml of N-methylmorpholine (NMM) (MW: 101.2, ρ: 0.92g / ml, 45mmol) and react for about 2 hours. The rutin peptide sequence is coupled with the corresponding Fmoc protection or Boc protection amino acid one by one, and the molar equivalent of each Fmoc protection or Boc pro...

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Abstract

The invention relates to a preparation method of bivalirudin, which is used for avoiding generation of impurities. The method comprises the following steps of: removing an Fmoc protecting group by using Fmoc-Leu-Wang resins or Fmoc-Leu-2-chlorotrityl resin as a raw material by means of solid phase coupling; coupling amino acid protected by the Fmoc one by one to obtain side-chain full-protection bivalirudin [9-20] peptide-resin; removing the protection of the Fmoc and performing solid phase coupling on Fmoc-Pro-Gly-Gly-Gly-Gly-OH pentapeptide segments to obtain a side-chain full-protection bivalirudin [4-20] peptide-resin; coupling the other coupled amino acid one by one to obtain a side-chain full-protection bivalirudin-resin; and splitting and removing resin and the protecting groups on the side-chain full-protection bivalirudin peptide-resins, and precipitating to obtain bivalirudin. According to the method, the impurities of bivalirudin [-Gly] and [+Gly] are reduced, and the yield and the purity of the product are improved.

Description

technical field [0001] The invention relates to an industrialized large-scale preparation method for bivalirudin, in particular to the preparation of bivalirudin by a solid-phase synthesis method, and belongs to the technical field of polypeptide synthesis. Background technique [0002] Bivalirudin is a synthetic anticoagulant drug whose anticoagulant component is a polypeptide with 20 amino acid residues at the C-terminus of a hirudin derivative. It was first developed by Biogen in Switzerland, and then transferred to Cambridge Medicines Company (The Medicines Company, TMC). Cambridge Medicines submitted a New Drug Application (NDA) to the FDA on December 23, 1997, until December 15, 2000. It has only been approved as an anticoagulant for patients with unstable angina undergoing percutaneous transluminal coronary angioplasty (PTCA). The marketed dosage form is powder injection, the trade name is Angiomax, and the specification is 250mg. [0003] Bivalirudin is a polypeptid...

Claims

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Application Information

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IPC IPC(8): C07K7/08C07K1/06C07K1/04C07K7/06
CPCY02P20/55
Inventor 田振平韩荣刚高永宏张道广彭海涛董佃强
Owner QILU PHARMA HAINAN
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