Preparation method of DOTATATE

A technology of -OH and resin, which is applied in the field of DOTATATE preparation, can solve the problems of unfavorable large-scale industrial production, difficult reaction control, unsuitable for industrial production, etc., and achieves easy control of the quality of linear peptides, cheap and easy-to-obtain starting materials, and inhibitory The effect of generation of impurities

Pending Publication Date: 2020-11-17
BEIJING SINOTAU INT PHARMA TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are few literatures about the preparation of DOTATATE, and the existing preparation methods of DOTATATE are cumbersome, the yield is low, the reaction is not easy to control, and the synthesis cost is high, which is not conducive to large-scale industrial production.
Due to the particularity of the DOTATATE structure, it is quite different from ordinary polypeptides. It has a ring structure, DOTA, and multiple unnatural amino acids. In the prior art, a large number of special reagents have been used, such as thallium trifluoroacetate (thallium trifluoroacetate ( Tl(TFA) 3 ), TCP-Resin, hydrazine hydrate, these reagents are costly, dangerous, difficult to control the reaction, unfriendly to the environment, and not suitable for industrial production
The TFA purification system method is used in the preparation of DOTATATE reported in the existing literature. After purification, there are still a large number of impurities, and the quality of DOTATATE cannot reach the pharmaceutical standard at all.
There is no research on the purification method of pharmaceutical grade DOTATATE in the prior art

Method used

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  • Preparation method of DOTATATE
  • Preparation method of DOTATATE
  • Preparation method of DOTATATE

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Step 1: Synthesis of DOTATATE Peptide Resin

[0026] 1) Synthesis of Fmoc-Thr(tBu)-Wang Resin: Weigh 210.02g of Wang Resin (the degree of substitution is 0.96mmol / g, the feeding scale is 201.62mmol), add it to the solid-phase synthesis column, and wash with DMF (1.30L) Once, drained, added DMF (1.30L) to swell for 30min, and drained the DMF.

[0027] Weigh Fmoc-Thr(tBu)-OH (240.43g) and HOBt (98.08g), dissolve with 0.30L DMF, freeze at 0-5°C for 30min, add DIC (114.52g) for activation for 3min; add the above-mentioned swelling resin to the activation solution , additional DMAP (7.40 g) and DMF (0.20 L) were added. Coupling time 4h. After coupling, wash 4 times with DMF (4*1.30L), wash 4 times with DCM (4*50ml), and dry at room temperature. Take the dried peptide resin, vacuum-dry (30min, 30°C, -0.98MPa), measure the degree of substitution of the resin, and the degree of substitution is 0.53mmol / g.

[0028] Wash all the Fmoc-Thr(tBu)-Wang Resin (the degree of substit...

Embodiment 2

[0035] Step 1: Synthesis of DOTATATE Peptide Resin

[0036] 1) Synthesis of Fmoc-Thr(tBu)-CTC Resin: Weigh 210.00g of 2-Cl-CTC resin (the degree of substitution is 0.97mmol / g, the feeding scale is 203.70mmol), add it to the solid-phase synthesis column, and use DMF ( 1.30L) was washed once, drained, added DMF (1.30L) to swell for 30min, and drained the DMF.

[0037] Fmoc-Thr(tBu)-OH (242.89g) was weighed and dissolved in 0.30L DMF, added to the above swelling resin, and then DIEA (157.96g) was added, and the coupling time was 6h. After coupling, wash 4 times with DMF (4*1.30L), wash 4 times with DCM (4*50ml), and dry at room temperature. Take the dried peptide resin, vacuum-dry (30min, 30°C, -0.98MPa), measure the degree of substitution of the resin, and the degree of substitution is 0.48mmol / g.

[0038] Wash all the Fmoc-Thr(tBu)-CTC Resin (the degree of substitution is 0.48mmol / g, the feeding scale is 121.68mmol) with DMF (1.30L), drained, add DMF (1.30L), CH 3 OH (13.04g...

Embodiment 3

[0049] Cleavage of DOTATATE peptide resin: Prepare TFA / TIS / MPR system lysate (1000ml, TFA / TIS / MPR=94 / 4 / 2) with TFA frozen at -10°C, stir well; 100g of the peptide resin obtained in 2 was slowly added to the above lysate, stirred, and cleaved at 20-30°C for 2 hours, the reaction solution was filtered with a dry sand core funnel, the resin was washed 3 times with TFA, the filtrate was collected, and the filtrate was distilled under reduced pressure (- 0.090MPa, 30.0°C, 40min); slowly add the remaining lysate (410ml) into the frozen, stirring isopropyl ether (4.1L), the solid precipitates, stir well after adding, let stand for 30min, separate the liquid, The lower layer was centrifuged, the solid was washed 4 times with isopropyl ether, and the solid was dried in vacuum (-0.98MPa, 30.0°C, 4h) to obtain a linear peptide (38.2g) with a HPLC purity of 55.8% and a yield of 106%.

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Abstract

The invention discloses a preparation method of DOTATATE. The method comprises the following specific steps of: sequentially coupling solid-phase carrier resin to amino acids, the N ends and side chains of which have protecting groups, according to the amino acid sequence of DOTATATE, and then, coupling DOTA(tBu)<3> so as to obtain linear peptide resin of DOTATATE; then, adding lysis solution to remove the protecting groups and cutting the resin to obtain linear peptide of DOTATATE; and finally, performing oxidization by using an oxidizing agent to obtain DOTATATE crude peptide. According to the preparation method in the invention, hydroxyl resin with easily available raw materials and low-price amino acids with side chain protection are adopted to synthesize the linear peptide resin; then, the linear peptide is cracked; the oxidizing agent easy to control is adopted to oxidize the linear peptide so as to obtain the target crude peptide; use of a highly toxic reagent is avoided; and finally, purification, salt conversion, concentration and drying are carried out, so that refined peptide is obtained. The method in the invention has the advantages of short synthesis route, easily controlled linear peptide quality, low synthesis cost, high yield and high finished product purity, and is suitable for large-scale production.

Description

technical field [0001] The invention belongs to the technical field of polypeptide synthesis, and in particular relates to a preparation method of DOTATATE. Background technique [0002] DOTATATE is the precursor of LUTETIUM DOTATATE LU-177, and LUTETIUM DOTATATE LU-177 can be obtained by chelating DOTATATE to LU-177. LUTETIUM DOTATATE LU-177 is a somatostatin-2 receptor analog with high affinity for somatostatin-2 receptors, it works by binding to cells bearing somatostatin receptors, the growth Inhibitin receptors may exist on some tumors. After binding to the receptors, the drug enters the cells and damages the tumor cells by radiation. It belongs to an emerging peptide receptor radionuclide therapy (PRTD). The drug was originally launched in Denmark in 2017. On January 26, 2018, ADVANCED ACCELERATOR APPLICATIONS USA was approved by the FDA. It is the first FDA-approved radiotherapy drug for the treatment of adult gastroenteropancreatic neuroendocrine tumors (GEP-NETs) ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/06C07K1/04C07K1/06
CPCC07K7/06Y02P20/55
Inventor 徐新盛
Owner BEIJING SINOTAU INT PHARMA TECH CO LTD
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