CN112940103A - Synthetic method of teriparatide impurity F - Google Patents
Synthetic method of teriparatide impurity F Download PDFInfo
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- CN112940103A CN112940103A CN201911259985.9A CN201911259985A CN112940103A CN 112940103 A CN112940103 A CN 112940103A CN 201911259985 A CN201911259985 A CN 201911259985A CN 112940103 A CN112940103 A CN 112940103A
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- fmoc
- impurity
- trt
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- teriparatide
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- 239000012535 impurity Substances 0.000 title claims abstract description 78
- 229960005460 teriparatide Drugs 0.000 title claims abstract description 37
- 108010049264 Teriparatide Proteins 0.000 title claims abstract description 33
- OGBMKVWORPGQRR-UMXFMPSGSA-N teriparatide Chemical compound C([C@H](NC(=O)[C@H](CCSC)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@@H](N)CO)C(C)C)[C@@H](C)CC)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC=1N=CNC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)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)N[C@@H](C(C)C)C(=O)N[C@@H](CC=1N=CNC=1)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CNC=N1 OGBMKVWORPGQRR-UMXFMPSGSA-N 0.000 title claims abstract description 32
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- AIISTODACBDQLW-WDSOQIARSA-N Trp-Leu-Arg Chemical compound C1=CC=C2C(C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O)=CNC2=C1 AIISTODACBDQLW-WDSOQIARSA-N 0.000 description 1
- BNQVUHQWZGTIBX-IUCAKERBSA-N Val-His Chemical compound CC(C)[C@H]([NH3+])C(=O)N[C@H](C([O-])=O)CC1=CN=CN1 BNQVUHQWZGTIBX-IUCAKERBSA-N 0.000 description 1
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- 125000003277 amino group Chemical group 0.000 description 1
- 235000012538 ammonium bicarbonate Nutrition 0.000 description 1
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- 108010062796 arginyllysine Proteins 0.000 description 1
- CXQGKOICHGQQMF-UHFFFAOYSA-N azido diphenyl phosphate Chemical compound C=1C=CC=CC=1OP(=O)(ON=[N+]=[N-])OC1=CC=CC=C1 CXQGKOICHGQQMF-UHFFFAOYSA-N 0.000 description 1
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Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/575—Hormones
- C07K14/635—Parathyroid hormone, i.e. parathormone; Parathyroid hormone-related peptides
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/04—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length on carriers
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/06—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length using protecting groups or activating agents
-
- 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
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- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Endocrinology (AREA)
- Gastroenterology & Hepatology (AREA)
- Zoology (AREA)
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- Analytical Chemistry (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention relates to the technical field of impurity synthesis, in particular to a preparation method of teriparatide impurity F. The invention discovers that teriparatide can generate asparagine impurities in the production and storage processes of teriparatide, named as impurities F, and the discovery and synthesis of the impurities are beneficial to the quality control of teriparatide bulk drugs; by adopting the synthetic method, the purity of the crude peptide can reach 42 percent, and the purity of the purified fine peptide can reach 92 percent. Compared with the prior art, the product obtained by the method has high purity, is easy to purify, and the yield is correspondingly improved.
Description
Technical Field
The invention relates to the technical field of impurity synthesis, in particular to a method for synthesizing teriparatide impurity F.
Background
Teriparatide (Teriparatide) is a fragment from position 1 to 34 of human parathyroid hormone, which has the same biological activity as human parathyroid hormone and is developed by Eli Lilly corporation of the united states for primary osteoporosis, hypogonadal osteoporosis and postmenopausal womenOsteoporosis in women. The peptide sequence is as follows: H-Ser-Val-Ser-Glu-Ile-Gln-Leu-Met-His-Asn-Leu-Gly-Lys-His-Leu-Asn-Ser-Met-Glu-Arg-Val-Glu-Trp-Leu-Arg-Lys-Lys-Leu-Gln-Asp30-Val31-His-Asn-Phe-OH。
Teriparatide produces various impurities during its production and storage, and the impurities currently reported to be found include Trp23(C ═ O) -teriparatide oxidation impurity, Trp23(OH) -teriparatide oxidation impurity, Met8(O) -teriparatide oxidation impurity, Met18(O) -teriparatide oxidation impurity, Met8,18(O) -teriparatide oxidation impurity, and the like. In order to ensure the safety and effectiveness of teriparatide drugs, the content of impurities in the product must be controlled. The discovery and synthesis of new impurities have important practical significance for the quality control of teriparatide bulk drugs.
Disclosure of Invention
In view of the above, the present invention provides a method for synthesizing teriparatide impurity F. The discovery and synthesis of the impurity are beneficial to the quality control of the teriparatide bulk drug; by adopting the synthetic method, the purity of the crude peptide can reach 42 percent, and the purity of the purified fine peptide can reach 92 percent.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides teriparatide impurity F, the structural formula of which is shown as formula I:
it was found during the development of the present application that during its production and storage an impurity of the aspartimide type, designated impurity F, i.e. 30 of the teriparatide residues, is produced#Asp and 31#And nucleophilic substitution reaction is carried out between Val to form a five-membered ring. The peptide sequence is as follows:
H-Ser-Val-Ser-Glu-Ile-Gln-Leu-Met-His-Asn-Leu-Gly-Lys-His-Leu-Asn-Ser-Met-Glu-Arg-Val-Glu-Trp-Leu-Arg-Lys-Lys-Leu-Gln-Asu-Val-His-Asn-Phe-OH. The amino acid sequence is shown as SEQ ID NO: 1 is shown.
The invention also provides a synthetic method of the teriparatide impurity F, which comprises the following steps:
step A: by adopting a solid-phase synthesis method, taking resin as a carrier, sequentially coupling Fmoc-Asn (Trt) -OH, Fmoc-His (Trt) -OH, Fmoc-Val-OH, Fmoc-Asp (ODmab) -OH, Fmoc-Gln (Trt) -OH, Fmoc-Leu-OH, Fmoc-Lys (Boc) -OH, Fmoc-Arg (Pbf) -OH, Fmoc-Leu-OH, Fmoc-Trp (Boc) -OH, Fmoc-Glu (OtBu) -OH, Fmoc-Val-OH, Fmoc-Arg (Pbf) -OH, Fmoc-Glu (OtBu) -OH, Fmoc-Met-OH, Fmoc-Ser (tBu) -OH, Fmoc-Asn (Trt) -OH, Fmoc-His (Trt, Fmoc-Lys (Boc) -OH, Fmoc-Gly-OH, Fmoc-Leu-OH, Fmoc-Asn (Trt) -OH, Fmoc-His (Trt) -OH, Fmoc-Met-OH, Fmoc-Leu-OH, Fmoc-Gln (Trt) -OH, Fmoc-Ile-OH, Fmoc-Glu (OtBu) -OH, Fmoc-Ser (tBu) -OH, Fmoc-Val-OH and Boc-Ser (tBu) -OH to obtain a peptide resin with a protecting group; the PG protecting group in Asp (PG) is an All or Dmab protecting group;
and B: removing PG protecting group of Asp (PG), and closing the ring to obtain peptide resin;
and C: the peptide resin is cracked to obtain teriparatide impurity F.
The purity of the target product obtained by the method reported by the inventor in the literature (DBU catalysis or acid degradation) is less than 1%, and the purity of the target product obtained by the conventional synthesis method (such as HOAt/PyAop/DIPEA ring closure) is only about 1%, so that the product with higher purity cannot be obtained. The invention provides a method for preparing teriparatide impurity F, and the mechanism is 30#The carboxyl exposed on the Asp side chain and diphenylphosphoryl azide generate acyl azide with higher activity, and then the acyl azide and the acyl azide react with 31#And performing nucleophilic substitution on the amino group of Val to obtain the target product. The product obtained by the method has high purity, is easy to purify, and the yield is correspondingly improved.
Preferably, in the step A, the resin is Wang resin, and the substitution degree is 0.1-3.0 mmol/g.
In the specific embodiment provided by the invention, the substitution degree of Wang Resinn is 0.5-0.8 mmol/g.
Preferably, in the step A, the coupling agent adopted for coupling is one or more of HOBt/DIPCDI, HOBt/PyBop/DIPEA, HBTU/HOBt/DIPEA, HOAt/DIPCDI, HATU/HOAt/DIPEA or HOAt/PyAop/DIPEA.
Preferably, in the step A, the reagent for removing the Fmoc protecting group used for coupling is 10-30% piperidine solution, and the solvent used for dissolving the amino acid and the piperidine solution is one or more of NMP, THF, DCM, DMF or DMSO.
Preferably, in step A, the Fmoc protecting group removing reagent used for coupling is a 20% piperidine solution.
Preferably, in step B, the PG protecting group in Asp (PG) is an All protecting group, the catalyst used for removing the protecting group is tetratriphenylphosphine palladium, and the scavenging agent is phenylsilane or morpholine.
Preferably, in step B, the PG protecting group in Asp (PG) is a Dmab protecting group, and the removing agent is 1-5% hydrazine hydrate-DMF solution.
Preferably, in step B, the PG protecting group in Asp (PG) is a Dmab protecting group, and the scavenger used for removal is 2% hydrazine hydrate-DMF solution.
Preferably, in the step B, the ring closing is performed by using diphenyl phosphorazidate.
Preferably, in step C, the lysis solution used for the lysis is a trifluoroacetic acid solution containing a capture reagent, wherein the capture reagent is PhSMe, PhOH, EDT, or H2One or more of O, TIS and PhOMe.
Preferably, the capture agent is TIS.
Preferably, the volume ratio of trifluoroacetic acid to TIS in the lysis solution is (90-99): (1-10).
Preferably, the volume ratio of trifluoroacetic acid to TIS in the lysate is 95: 5.
the invention provides a synthetic method of teriparatide impurity F. The structural formula of the teriparatide impurity F is shown as a formula I. The invention has the following advantages:
the present invention found that teriparatide produced asparagine impurities, designated impurity F, i.e. 30 of the teriparatide residues during its production and storage#Asp and 31#And nucleophilic substitution reaction is carried out between Val to form a five-membered ring. The discovery and synthesis of the impurity are beneficial to the quality control of the teriparatide bulk drug;
the purity of the obtained target product is less than 1% by adopting a method reported in the literature (DBU catalysis or acid degradation), and the purity of the obtained target product is only about 1% by adopting a conventional synthesis method (HOAt/PyAop/DIPEA ring closure), so that a product with higher purity cannot be obtained; by adopting the method, the purity of the crude peptide can reach 42 percent, and the purity of the purified fine peptide can reach 92 percent. Compared with the prior art, the product obtained by the method has high purity, is easy to purify, and the yield is correspondingly improved.
Drawings
FIG. 1: synthetic scheme for impurity F;
FIG. 2: HPLC profile of crude peptide of impurity F (example 3);
FIG. 3: HPLC profile of fine peptide of impurity F (example 8);
FIG. 4: a primary mass spectrum of impurity F;
FIG. 5: a first-order mass spectrogram after enzyme digestion of impurity F;
FIG. 6: a secondary mass spectrogram of a fragment of which the parent ion M/Z is 1455 after the enzyme digestion of the impurity F;
FIG. 7: a secondary mass spectrogram of a fragment with a parent ion M/Z of 886 after the enzyme digestion of the impurity F;
FIG. 8: a secondary mass spectrogram of a fragment with parent ion M/Z of 702 after the enzyme digestion of the impurity F;
FIG. 9: a secondary mass spectrogram of a fragment with a parent ion M/Z of 872 after the enzyme digestion of the impurity F;
FIG. 10: the parent ion M/Z after the enzyme digestion of the impurity F is the comparison condition of the actually measured secondary fragment peak of 1455 sections and the theoretical secondary fragment of the target peptide sequence;
FIG. 11: the M/Z of parent ion after the enzyme digestion of the impurity F is 886, and the comparison condition of the actually measured secondary fragment peak and the theoretical secondary fragment of the target peptide sequence is obtained;
FIG. 12: comparing the actual measurement secondary fragment peak of the fragment with the parent ion M/Z of 702 after the enzyme digestion of the impurity F with the theoretical secondary fragment of the target peptide sequence;
FIG. 13: and (3) comparing the actually measured secondary fragment peak of the fragment with the 872 parent ion M/Z of the fragment after the enzyme digestion of the impurity F with the theoretical secondary fragment of the target peptide sequence.
Detailed Description
The invention discloses a method for synthesizing teriparatide impurity F, which can be realized by appropriately improving process parameters by referring to the content. It is expressly intended that all such similar substitutes and modifications which would be obvious to one skilled in the art are deemed to be included in the invention. While the methods and applications of this invention have been described in terms of preferred embodiments, it will be apparent to those of ordinary skill in the art that variations and modifications in the methods and applications described herein, as well as other suitable variations and combinations, may be made to implement and use the techniques of this invention without departing from the spirit and scope of the invention.
Abbreviations and english means as follows:
the method for specifically preparing teriparatide impurity F provided by the invention comprises the following steps:
1. using Wang Resin as a carrier, the substitution degree is 0.1-3.0 mmol/g, and sequentially coupling and protecting amino acids from the C end to the N end according to the peptide sequence according to a polypeptide solid phase synthesis method, wherein the substitution degree is 30#Asp adopts Fmoc-Asp (PG) -OH, PG is an All or Dmab protecting group, 1#The Ser is Boc-Ser (tBu) -OH, and other residues are conventional amino acids. Sequentially coupling one or more of HOBt/DIPCDI, HOBt/PyBop/DIPEA, HBTU/HOBt/DIPEA, HOAt/DIPCDI, HATU/HOAt/DIPEA and HOAt/PyAop/DIPEA by using coupling agents of Fmoc-Asn (Trt) -OH, Fmoc-His (Trt) -OH, Fmoc-Val-OH, Fmoc-Asp (PG) -OH, Fmoc-Gln (Trt) -OH, Fmoc-Leu-OH, Fmoc-Lys (Boc) -OH, Fmoc-Arg (Pbf) -OH, Fmoc-Leu-OH, Fmoc-Trp (Boc) -OH, Fmoc-Glu (OtBu) -OH, Fmoc-Val-OH, Fmoc-Arg (f) -OH, Fmoc-Leu-OH, Fmoc-Glu (Boc) -OH, Fmoc-Glu-OH, Fmoc-Ser (tBu) -OH, Fmoc-Asn (Trt) -OH, Fmoc-Leu-OH, Fmoc-His (Trt) -OH, Fmoc-Lys (Boc) -OH, Fmoc-Gly-OH, Fmoc-Leu-OH, Fmoc-Asn (Trt) -OH, Fmoc-His (Trt) -OH, Fmoc-Met-OH, Fmoc-Leu-OH, Fmoc-Gln (Trt) -OH, Fmoc-Ile-OH, Fmoc-Ser (tBu) -OH, Fmoc-Lys (Trt) -OH, Fmoc-Leu-OH, Fmoc-Lys (Troc-Glu (OtBu) -OH, Fmoc-Ser (tBu) -OH, Fmoc-Val-OH and Boc-Ser (tBu) -OH. PG is an All or Dmab protecting group, a reagent for removing the Fmoc protecting group is 20% piperidine solution, and a solvent used for dissolving the amino acid and the 20% piperidine solution is one or more of NMP, THF, DCM, DMF and DMSO.
2. Removing PG protective group, if PG is All, adopting palladium tetratriphenylphosphine for catalysis, and taking phenylsilane or morpholine as a scavenging agent for removal; if PG is Dmab, it is removed by 2% hydrazine hydrate-DMF solution.
3. Firstly, diphenyl phosphorazide is adopted to close a ring, and then trifluoroacetic acid solution containing a trapping agent is adopted to crack to obtain impurities F, wherein the trapping agent is PhSMe, PhOH, EDT and H2One or more of O, TIS, PhOMe, more preferably, the lysate is a TFA/TIS composition, wherein the TFA, TIS are present in a volume ratio of 95: 5.
the specific route is shown in figure 1.
The teriparatide impurity F and the reagent or instrument used in the preparation method thereof provided by the invention can be purchased from the market.
The invention is further illustrated by the following examples:
example 1: coupling of amino acids
62.5g (50mmol) of Wang Resin with a substitution degree of 0.8mmol/g was weighed, charged into a solid phase reaction column, washed 2 times with DMF, after swelling the Resin with DMF for 30 minutes, 19.37g (50mmol) of Fmoc-Phe-OH, 8.1g (60mmol) of HOBt and 6.1g (5mmol) of DMAP were weighed, dissolved in DMF, 8.2g (65mmol) of DIPCDI was added under ice bath, charged into a solid phase reaction column, reacted at room temperature for 1 hour, and washed 6 times with DMF. A mixture of 79.1g (1000mmol) pyridine and 102.1g (1000mmol) acetic anhydride was added to block the Resin for 6 hours, washed with DMF for 6 times, and the methanol was then contracted and drained to give 71.4g Fmoc-Phe-Wang Resin with a degree of detection of substitution of 0.3 mmol/g.
The Fmoc protecting group was removed in 20% piperidine (reaction time 5+7 min) and washed 6 times with DMF. Sequentially coupling Fmoc-Asn (Trt) -OH, Fmoc-His (Trt) -OH, Fmoc-Val-OH, Fmoc-Asp (OAll) -OH, Fmoc-Gln (Trt) -OH, Fmoc-Lys (Boc) -OH, Fmoc-Arg (Pbf) -OH, Fmoc-Leu-OH, Fmoc-Trp (Boc) -OH, Fmoc-Glu (Glu-Glu) -OH, (Fmoc-Arg-OH, (Pbf) -OH, Fmoc-Leu-OH, Fmoc-Trp (Boc) -OH, Fmoc-Glu (Glu) -OH, Fmoc-Val-OH, (Met-Arg-OH), (Fmoc-f) -OH, Fmoc-OH, OtBu-Glu-OH, Fmoc-Glu (Met-Trp) (Boc) -OH, Fmoc-Glu-OH, Fmoc-Glu (Boc, Fmoc-Ser (tBu) -OH, Fmoc-Asn (Trt) -OH, Fmoc-Leu-OH, Fmoc-His (Trt) -OH, Fmoc-Lys (Boc) -OH, Fmoc-Gly-OH, Fmoc-Leu-OH, Fmoc-Asn (Trt) -OH, Fmoc-His (Trt) -OH, Fmoc-Met-OH, Fmoc-Leu-OH, Fmoc-Gln (Trt) -OH, Fmoc-Ile-OH, Fmoc-Glu (OtBu) -OH, Fmoc-Ser (tBu) -OH, Fmoc-Val-OH and Boc-Ser (tBu) -OH.
Example 2: coupling of amino acids
100.0g (50mmol) of Wang Resin with a substitution degree of 0.5mmol/g was weighed, charged into a solid phase reaction column, washed 2 times with DMF, after swelling the Resin with DMF for 30 minutes, 38.74g (100mmol) of Fmoc-Phe-OH, 16.2g (120mmol) of HOBt and 52.1g (100mmol) of PyBOP were weighed out dissolved in DMF, 25.9g (200mmol) of DIPEA was added under ice bath, charged into a solid phase reaction column, reacted at room temperature for 2 hours, and washed 6 times with DMF. A further mixture of 79.1g (1000mmol) pyridine and 102.1g (1000mmol) acetic anhydride was added to block the Resin for 6 hours, washed 6 times with DMF and methanol contracted and drained to give 115.4g Fmoc-Phe-Wang Resin with a degree of detection of substitution of 0.26 mmol/g.
The Fmoc protecting group was removed in 20% piperidine (reaction time 5+7 min) and washed 6 times with DMF. Sequentially coupling Fmoc-Asn (Trt) -OH, Fmoc-His (Trt) -OH, Fmoc-Val-OH, Fmoc-Asp (ODmab) -OH, Fmoc-Gln (Trt) -OH, Fmoc-Leu-OH, Fmoc-Lys (Boc) -OH, Fmoc-Arg (Pbf) -OH, Fmoc-Leu-OH, Fmoc-Trp (Boc) -OH, Fmoc-Glu (Glu-Glu) -OH, (Fmoc-Val-OH, Fmoc-Arg (Pbf) -OH, Fmoc-Leu-OH, Fmoc-Trp (Boc) -OH, Fmoc-Glu-OtBu) -OH, Fmoc-Val-OH, (Fmoc-Arg-f) -OH, Fmoc-Glu-OH, OtBu-OH, Fmoc-Glu-OH, Fmoc-Arg (Boc) -OH, Fmoc-Glu-OH, Fmoc-Glu, Fmoc-Ser (tBu) -OH, Fmoc-Asn (Trt) -OH, Fmoc-Leu-OH, Fmoc-His (Trt) -OH, Fmoc-Lys (Boc) -OH, Fmoc-Gly-OH, Fmoc-Leu-OH, Fmoc-Asn (Trt) -OH, Fmoc-His (Trt) -OH, Fmoc-Met-OH, Fmoc-Leu-OH, Fmoc-Gln (Trt) -OH, Fmoc-Ile-OH, Fmoc-Glu (OtBu) -OH, Fmoc-Ser (tBu) -OH, Fmoc-Val-OH and Boc-Ser (tBu) -OH.
Example 3: synthesis of impurity F
12.4g (0.5eq) of palladium tetratriphenylphosphine, 23.2g (10eq) of phenylsilane, and 500mL of methylene chloride were charged into the reaction column of example 1, and the mixture was reacted at room temperature for 1 hour, after which the reaction solution was taken out, washed with methylene chloride six times, washed with tetrahydrofuran three times, transferred to a three-necked flask, charged with 43.4g (20eq) of triethylamine, 117.9g (20eq) of diphenyl phosphorazidate, and 500mL of tetrahydrofuran, and heated to reflux for 8 hours, cooled to room temperature, filtered, washed with tetrahydrofuran six times, contracted three times with methyl t-butyl ether, and vacuum-dried to obtain 140g of a peptide resin.
Adding lysis solution (TFA: TIS 95:5, volume ratio) 1.2L, splitting at room temperature for 2 hours, filtering, pouring the filtrate into 12L of ether for sedimentation, centrifuging, washing, and drying in vacuum to obtain crude F peptide impurity 90g with purity of 42.96% (figure 2, table 1).
TABLE 1
Example 4: synthesis of impurity F
A2% hydrazine hydrate-DMF solution is added into the peptide resin reaction column of the example 2 to remove Dmab protecting groups (reaction time is 10+10 minutes), DMF is washed for 6 times, tetrahydrofuran is washed for three times, the mixture is transferred into a three-neck flask, 30.4g (10eq) of triethylamine, 82.6g (10eq) of azido diphenyl phosphate and 1L of tetrahydrofuran are added, the mixture is heated to reflux for reaction for 15 hours, the mixture is cooled to room temperature, filtered, washed for six times by the tetrahydrofuran, methyl tert-butyl ether is contracted for three times, and vacuum drying is carried out to obtain 200g of peptide resin.
Adding lysis solution (TFA: H)2O95: 5 by volume) 1.6L room temperature for 2 h, filtered, the filtrate was poured into 16L ether for sedimentation, washed by centrifugation and dried in vacuo to give 120g of crude peptide F as impurity with a purity of 40.35%.
Example 5: synthesis of impurity F
1/10 of the peptide resin of example 1 was weighed, 1.2g (0.5eq) of palladium tetratriphenylphosphine, 2.3g (10eq) of phenylsilane and 50mL of dichloromethane were added, the mixture was reacted at room temperature for 1 hour, the reaction solution was removed, dichloromethane was washed six times, 1.6g (12mmol) of HOAt and 5.2g (10mmol) of PyAOP were further weighed and dissolved in DMF, 2.6g (20mmol) of DIPEA was added under ice bath, the mixture was put into a solid phase reaction column, reacted at room temperature for 15 hours, the reaction solution was removed, DMF was washed six times, methyl t-butyl ether was contracted three times, and vacuum-dried to obtain 15g of the peptide resin. Adding a lysis solution (TFA: TIS: 95:5, volume ratio) 150mL, splitting at room temperature for 2 hours, filtering, pouring the filtrate into 1.5L of diethyl ether for sedimentation, centrifugally washing, and drying in vacuum to obtain crude peptide 9g of impurity F with the purity of about 1%.
Example 6: synthesis of impurity F
Taking 5g of teriparatide peptide resin, adding 50% DBU-DMF solution, reacting for 7d at room temperature, pumping out reaction liquid, washing for six times by DMF, shrinking for three times by methyl tert-butyl ether, drying in vacuum, adding 50mL of lysate (TFA: TIS ═ 95:5, volume ratio) for 2 hours at room temperature, filtering, pouring filtrate into 500mL of diethyl ether for settling, centrifuging, washing, and drying in vacuum to obtain 2g of impurity F crude peptide with the purity of 0.4%.
Example 7: synthesis of impurity F
Dissolving 0.1g of teriparatide refined peptide in 5mL of purified water, adjusting the pH value to 3 by using dilute hydrochloric acid, heating to 90 ℃ for reaction for 3d, and detecting the purity of the impurity F by HPLC (high performance liquid chromatography) to be 0.8%.
Example 8: purification of impurity F
The impurity F obtained in example 2 was purified by HPLC with a sample loading of 30 g/time, fractions were collected stepwise according to the chromatographic peak starting from about 10% of the beginning of the main peak and stepwise according to the chromatographic peak, and the collection was stopped when the main peak dropped to about 10%. The fractions were concentrated by rotary evaporation and lyophilized to give purified F peptide 15.5g with a purity of 92.13% (FIG. 3, Table 2).
1. HPLC preparative purification chromatography conditions were as follows:
(1) mobile phase a 1: 0.1% TFA solution, mobile phase B1: pure acetonitrile;
(2) novasep LC150 chromatography system, 15cm chromatography column packed Welch Ultimate HS-C188 μmPacking (the theoretical plate number of column effective value is not less than 5000);
(3) monitoring wavelength: 220 nm;
(4) flow rate: 500mL/min
(5) Purification gradient:
time (min) | Mobile phase a1 (%) | Mobile phase B1 (%) |
0 | 90 | 10 |
5 | 75 | 25 |
105 | 65 | 35 |
115 | 30 | 70 |
2. The chromatographic conditions for HPLC analysis were as follows:
(1) mobile phase: phase A: acetonitrile: ammonium sulfate buffer 10: 90 (V: V), phase B: acetonitrile: ammonium sulfate buffer 50: 50 (V: V);
(2) a chromatographic column: waters BEHPeptide 300-C18-1.7 μm 2.1 x 100 mm;
(3) flow rate: 0.4 mL/min;
(4) monitoring wavelength: 214 nm;
(5) column temperature: 60 ℃;
(6) detecting a gradient:
time (min) | Mobile phase A (%) | Mobile phase B (%) |
0 | 100 | 0 |
5 | 65 | 35 |
29 | 61 | 39 |
30 | 0 | 100 |
35 | 0 | 100 |
35.1 | 100 | 0 |
40 | 100 | 0 |
TABLE 2
Example 9: confirmation of the Structure of impurity F
In order to determine the structure of the impurity F, primary mass spectrometry is carried out on the impurity F, treatment is carried out by Trypsin enzyme, and the primary mass spectrometry and the secondary mass spectrometry after enzyme digestion are analyzed.
1. The experimental conditions are as follows:
(1) the instrument model is as follows: MALDI-TOF-TOF, Autoflex Speed;
(2) first order mass spectrum
Laser wavelength and frequency: ND is YAG 355nm, 1000 Hz;
polarity: positive;
the operation mode is as follows: RP _ 700-;
detector voltage: 2000V;
analysis software: FlexAnalysis;
target type: MTP 384 ground steel;
matrix: DHB is dissolved to 20mg/mL by 30% acetonitrile water solution;
matrix and sample: mixing according to the volume ratio of 1:1, naturally drying, and placing into a mass spectrometry test chamber.
(3) Second order mass spectrum
Laser wavelength and frequency: ND is YAG 355nm, 1000 Hz;
polarity: a Positive reflector;
the operation mode is as follows: LIFT;
detector voltage: 2000V;
analysis software: flexaanalysts, BioTools, Sequence Editor;
target type: MTP 384 ground steel;
matrix: DHB is dissolved to 20mg/mL by 30% acetonitrile water solution;
matrix and sample: mixing according to the volume ratio of 1:1, naturally drying, and placing into a mass spectrometry test chamber.
(4) Treatment of the sample before enzyme digestion: the purified peptide F as an impurity was dissolved in 0.1% TFA to give an aqueous solution having a concentration of about 0.1mg/mL, and the solution was subjected to primary mass spectrometry.
(5) And (3) processing an enzyme digestion sample: dissolving the impurity F fine peptide into aqueous solution with the concentration of about 1mg/mL by using 50mM ammonium bicarbonate solution, carrying out enzyme digestion on the aqueous solution by using Trypsin, and analyzing the primary and secondary mass spectrums after the enzyme digestion.
(6) The results are as follows:
1) the first mass spectrum of impurity F is shown in fig. 4.
2) And the result of the primary mass spectrum after the enzyme digestion of the impurity F is shown in figure 5.
3) And the results of the secondary mass spectrometry after the enzyme digestion of the impurity F are shown in FIGS. 6-9.
4) The results of MALDI-TOF-TOF second mass spectrometry using Biotools software are shown in FIGS. 10-13.
(7) Data analysis
Table 3: comparing the actually measured first-class mass spectrum of the impurity F with a theoretical value
Sample (I) | Theoretical value of first-order mass spectrum | First-order mass spectrum measured value |
Impurity F | 4100.727 | 4100.824 |
Table 4: comparing the actual measurement primary mass spectrum after the enzyme digestion of the impurity F with a theoretical value
Fragment peptide sequences | Theoretical value of first-order mass spectrum | First-order mass spectrum measured value |
SVSEIQLMHNLGK | 1455.762 | 1455.732 |
HLNSMER | 886.420 | 886.461 |
VEWLR | 702.393 | 702.387 |
LQDVHNF | 872.426 | 872.406 |
A. As can be seen from Table 3, the first mass spectrum of impurity F is consistent with the theoretical value.
B. Trypsin can selectively hydrolyze carboxyl-terminal peptide bonds of lysine or arginine in protein, and it can be seen from Table 4 that the secondary fragment peaks of peptide fragments after the enzyme digestion of impurity F are consistent with theory, and the secondary fragment peaks of peptide fragments with parent ions M/Z of 1455, 886, 702 and 872 are respectively consistent with the theoretical fragment peak of peptide fragment SVSEIQLMHNLGK, HLNSMER, VEWLR, LQDVHNF, wherein the LQDVHNF peptide fragment is obtained by hydrolyzing LQ (Asu) VHNF in the enzyme digestion process.
Thus, the peptide sequence of impurity F can be determined as: H-Ser-Val-Ser-Glu-Ile-Gln-Leu-Met-His-Asn-Leu-Gly-Lys-His-Leu-Asn-Ser-Met-Glu-Arg-Val-Glu-Trp-Leu-Arg-Lys-Lys-Leu-Gln-Asu-Val-His-Asn-Phe-OH (SEQ ID NO: 1).
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Sequence listing
<110> Shenzhen Hanyu pharmaceutical stockings Limited
<120> synthetic method of teriparatide impurity F
<130> S19P3273
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 34
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<220>
<221> UNSURE
<222> (30)..(30)
<223> Xaa(30)=Asu
<220>
<221> UNSURE
<222> (30)..(30)
<223> The 'Xaa' at location 30 stands for Gln, Arg, Pro, or Leu.
<400> 1
Ser Val Ser Glu Ile Gln Leu Met His Asn Leu Gly Lys His Leu Asn
1 5 10 15
Ser Met Glu Arg Val Glu Trp Leu Arg Lys Lys Leu Gln Xaa Val His
20 25 30
Asn Phe
Claims (9)
1. A method for synthesizing teriparatide impurity F is characterized in that the structural formula of the teriparatide impurity F is shown as a formula I:
the synthesis method comprises the following steps:
step A: by adopting a solid-phase synthesis method, taking resin as a carrier, sequentially coupling Fmoc-Asn (Trt) -OH, Fmoc-His (Trt) -OH, Fmoc-Val-OH, Fmoc-Asp (ODmab) -OH, Fmoc-Gln (Trt) -OH, Fmoc-Leu-OH, Fmoc-Lys (Boc) -OH, Fmoc-Arg (Pbf) -OH, Fmoc-Leu-OH, Fmoc-Trp (Boc) -OH, Fmoc-Glu (OtBu) -OH, Fmoc-Val-OH, Fmoc-Arg (Pbf) -OH, Fmoc-Glu (OtBu) -OH, Fmoc-Met-OH, Fmoc-Ser (tBu) -OH, Fmoc-Asn (Trt) -OH, Fmoc-His (Trt, Fmoc-Lys (Boc) -OH, Fmoc-Gly-OH, Fmoc-Leu-OH, Fmoc-Asn (Trt) -OH, Fmoc-His (Trt) -OH, Fmoc-Met-OH, Fmoc-Leu-OH, Fmoc-Gln (Trt) -OH, Fmoc-Ile-OH, Fmoc-Glu (OtBu) -OH, Fmoc-Ser (tBu) -OH, Fmoc-Val-OH and Boc-Ser (tBu) -OH to obtain a peptide resin with a protecting group; the PG protecting group in Asp (PG) is an All or Dmab protecting group;
and B: removing PG protecting group of Asp (PG), and closing the ring to obtain peptide resin;
and C: the peptide resin is cracked to obtain teriparatide impurity F.
2. The synthetic method according to claim 1, wherein in the step A, the resin is Wang resin, and the substitution degree is 0.1-3.0 mmol/g.
3. The synthetic method of claim 1, wherein in step a, the coupling agent used for coupling is one or more of HOBt/dipdi, HOBt/PyBop/DIPEA, HBTU/HOBt/DIPEA, HOAt/DIPCDI, HATU/HOAt/DIPEA, or HOAt/PyAop/DIPEA.
4. The synthesis method of claim 1, wherein in the step A, the reagent for removing the Fmoc protecting group used in the coupling is a 10-30% piperidine solution, and the solvent used for dissolving the amino acid and the piperidine solution is one or more of NMP, THF, DCM, DMF or DMSO.
5. The synthesis method according to claim 1, wherein in step B, the PG protecting group in Asp (PG) is an All protecting group, the catalyst used for the removal is palladium tetratriphenylphosphine, and the scavenger is phenylsilane or morpholine.
6. The synthetic method according to claim 1, wherein in step B, the PG protecting group in asp (PG) is a Dmab protecting group, and the removing agent is 1-5% hydrazine hydrate-DMF solution.
7. The synthesis method of claim 1, wherein in step B, the closed ring is closed by diphenyl phosphorazidate.
8. The synthesis method according to any one of claims 1 to 7, characterized in that in step C, the lysis solution used for the lysis is a trifluoroacetic acid solution containing a capture agent, wherein the capture agent is PhSMe, PhOH, EDT, H2One or more of O, TIS and PhOMe.
9. The synthesis method according to claim 8, wherein the capture agent is TIS, and the volume ratio of trifluoroacetic acid to TIS in the lysate is (90-99): (1-10).
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Citations (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20080287650A1 (en) * | 2007-03-01 | 2008-11-20 | Avi Tovi | High purity peptides |
CN103467595A (en) * | 2013-09-06 | 2013-12-25 | 深圳翰宇药业股份有限公司 | Method for preparing teriparatide |
CN104017064A (en) * | 2014-06-13 | 2014-09-03 | 杭州诺泰制药技术有限公司 | Method for preparing teriparatide |
EP3000821A1 (en) * | 2014-09-25 | 2016-03-30 | Uniwersytet Wroclawski | Pseudofactin derivatives, a method of synthesizing pseudofactin derivatives and their use |
WO2017114382A1 (en) * | 2015-12-31 | 2017-07-06 | 深圳翰宇药业股份有限公司 | Method for synthesizing c-terminal modified peptide |
CN107383171A (en) * | 2017-08-15 | 2017-11-24 | 苏州科技大学 | A kind of method by secondary cyclization synthesis in solid state Pu Kana peptides |
US20210024579A1 (en) * | 2018-03-28 | 2021-01-28 | University Of Warwick | Macrocyclization of peptidomimetics |
Family Cites Families (3)
Publication number | Priority date | Publication date | Assignee | Title |
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EP1557176A1 (en) * | 2004-01-26 | 2005-07-27 | Ferring B.V. | Treatment of bone metastases by means of PTH receptor agonists |
US20060069021A1 (en) * | 2004-08-13 | 2006-03-30 | Nastech Pharmaceutical Company Inc. | Compositions and methods for intranasal administration of inactive analogs of PTH or inactivated preparations of PTH or PTH analogs |
CN104530218A (en) * | 2015-01-07 | 2015-04-22 | 哈尔滨吉象隆生物技术有限公司 | Solid-phase synthesis method of teriparatide |
-
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Patent Citations (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20080287650A1 (en) * | 2007-03-01 | 2008-11-20 | Avi Tovi | High purity peptides |
CN103467595A (en) * | 2013-09-06 | 2013-12-25 | 深圳翰宇药业股份有限公司 | Method for preparing teriparatide |
CN104017064A (en) * | 2014-06-13 | 2014-09-03 | 杭州诺泰制药技术有限公司 | Method for preparing teriparatide |
EP3000821A1 (en) * | 2014-09-25 | 2016-03-30 | Uniwersytet Wroclawski | Pseudofactin derivatives, a method of synthesizing pseudofactin derivatives and their use |
WO2017114382A1 (en) * | 2015-12-31 | 2017-07-06 | 深圳翰宇药业股份有限公司 | Method for synthesizing c-terminal modified peptide |
CN107383171A (en) * | 2017-08-15 | 2017-11-24 | 苏州科技大学 | A kind of method by secondary cyclization synthesis in solid state Pu Kana peptides |
US20210024579A1 (en) * | 2018-03-28 | 2021-01-28 | University Of Warwick | Macrocyclization of peptidomimetics |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN116120426A (en) * | 2022-12-29 | 2023-05-16 | 江苏诺泰澳赛诺生物制药股份有限公司 | Method for purifying liraglutide oxidized impurities |
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