CN111574594A - Preparation method of Bulevirtide - Google Patents
Preparation method of Bulevirtide Download PDFInfo
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- CN111574594A CN111574594A CN202010457818.1A CN202010457818A CN111574594A CN 111574594 A CN111574594 A CN 111574594A CN 202010457818 A CN202010457818 A CN 202010457818A CN 111574594 A CN111574594 A CN 111574594A
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- 238000002360 preparation method Methods 0.000 title claims abstract description 18
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- JDDWRLPTKIOUOF-UHFFFAOYSA-N 9h-fluoren-9-ylmethyl n-[[4-[2-[bis(4-methylphenyl)methylamino]-2-oxoethoxy]phenyl]-(2,4-dimethoxyphenyl)methyl]carbamate Chemical compound COC1=CC(OC)=CC=C1C(C=1C=CC(OCC(=O)NC(C=2C=CC(C)=CC=2)C=2C=CC(C)=CC=2)=CC=1)NC(=O)OCC1C2=CC=CC=C2C2=CC=CC=C21 JDDWRLPTKIOUOF-UHFFFAOYSA-N 0.000 claims description 4
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- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 description 2
- AGPKZVBTJJNPAG-WHFBIAKZSA-N L-isoleucine Chemical compound CC[C@H](C)[C@H](N)C(O)=O AGPKZVBTJJNPAG-WHFBIAKZSA-N 0.000 description 2
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- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 2
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- 108091022863 bile acid binding Proteins 0.000 description 1
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- ZTUXEFFFLOVXQE-UHFFFAOYSA-N tetradecanoic acid Chemical compound CCCCCCCCCCCCCC(O)=O.CCCCCCCCCCCCCC(O)=O ZTUXEFFFLOVXQE-UHFFFAOYSA-N 0.000 description 1
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- OHSJPLSEQNCRLW-UHFFFAOYSA-N triphenylmethyl radical Chemical compound C1=CC=CC=C1[C](C=1C=CC=CC=1)C1=CC=CC=C1 OHSJPLSEQNCRLW-UHFFFAOYSA-N 0.000 description 1
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Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/001—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof by chemical synthesis
-
- 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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- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Biochemistry (AREA)
- Gastroenterology & Hepatology (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Chemical & Material Sciences (AREA)
- Biophysics (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
Abstract
The invention provides a preparation method of Bulevirtide, which adopts a special protective amino acid fragment of Myristonyn-Gly-Thr (tBu) -Asn (Trt) -Leu-Ser (tBu) -Val-Pro-Asn (Trt) -Pro-Leu-Gly-Phe-Phe-Pro-OH, shortens the preparation process period and improves the total yield of products in large-scale preparation.
Description
Technical Field
The invention belongs to the technical field of preparation methods of polypeptide medicaments, and particularly relates to a preparation method of Bulevirtide.
Background
A new generation of oral direct antiviral drugs achieves a cure for hepatitis C, but chronic Hepatitis B (HBV) infected patients currently require long-term or even lifetime treatment. There are about 20 billion HBV infected patients all over the world, but most of them are acute infections, and the immune system of the human body can rapidly play a role to completely eliminate the HBV. In addition, 3.8 million infected patients are chronic and the virus is hidden in tissues and organs and cannot be completely eliminated by the immune system and drugs.
Researchers have found that a protein called liver bile acid transporter (NTCP, sodium taurocholate cotransporter polypeptide) on the surface of hepatocytes can specifically interact with the key receptor binding domain of HBV envelope proteins, which are receptors required for HBV infection of host cells. Therefore, blocking NTCP would be expected to cure hepatitis B. Bulevirtide is a novel Firstinclass hepatitis B medicament targeting NTCP.
Active results of the Bulevirtide phase IIb study were reported by MyrPharma at the annual meeting of the European Association for liver disease. And (3) displaying data: after 48 weeks of Bulevilide in combination with PEG-IFN alpha, more than 50% of HBV/HDV co-infected patients had no detectable HDVRNA. Also, hepatitis B surface antigen (HBsAg) in some of the subjects reached undetectable levels. More importantly, the effect can still be continued after the medicine is stopped, which also means that the bullevirtide is expected to be an option for curing hepatitis B in the future.
Bulevirtide qualifies for orphan medication awarded by the FDA and EMA for the treatment of HDV infection, is under the priority drug (PRIME) program by EMA, and has received FDA ' breakthrough therapy ' approval and United kingdom drug and health product administration (MHRA) ' approval of highly desirable innovative drugs (PIMs).
Bulevilide has the following structure:
Myristony-Gly-Thr-Asn-Leu5-Ser-Val-Pro-Asn-Pro10-Leu-Gly-Phe-Phe-Pro15-Asp-His-Gln-Leu-Asp20-Pro-Ala-Phe-Gly-Ala25-Asn-Ser-Asn-Asn-Pro30-Asp-Trp-Asp-Phe-Asn35-Pro-Asn-Lys-Asp-His40-Trp-Pro-Glu-Ala-Asn45-Lys-Val-Gly-NH2
the invention provides an efficient Bulevirtide preparation method, which improves the purity and yield of high products so as to meet the medical application.
Disclosure of Invention
The invention provides a novel high-efficiency preparation method, which adopts special protected amino acid, shortens the preparation process period and improves the product purity and yield in a large-scale preparation method.
The invention provides a preparation method of Bulevirtide, which comprises the following steps: amino resin is adopted as starting resin, the Bulevilide is prepared by a solid-phase polypeptide synthesis method, Bulevilide resin is obtained by the polypeptide solid-phase synthesis method, a Bulevilide crude product is obtained by acidolysis of the Bulevilide resin, and finally the Bulevilide crude product is purified to obtain a Bulevilide pure product.
Wherein, in addition to other conventional protected amino acids, the following special protected amino acid fragments are used in the synthesis process of the Bulevilide multi-resin:
Myristony-Gly-Thr(tBu)-Asn(Trt)-Leu-Ser(tBu)-Val-Pro-Asn(Trt)-Pro-Leu-Gly-Phe-Phe-Pro-OH
bulevilide peptide resin:
Myristony-Gly-Thr(tBu)-Asn(Trt)-Leu5-Ser(tBu)-Val-Pro-Asn(Trt)-Pro10-Leu-Gly-Phe-Phe-Pro15-Asp(OtBu)-His(Trt)-Gln(Trt)-Leu-Asp(OtBu)20-Pro-Ala-Phe-Gly-Ala25-Asn(Trt)-Ser(tBu)-Asn(Trt)-Asn(Trt)-Pro30-Asp(OtBu)-Trp(Boc)-Asp(OtBu)-Phe-Asn(Trt)35-Pro-Asn(Trt)-Lys(Boc)-Asp(OtBu)-His(Trt)40-Trp(Boc)-Pro-Glu(OtBu)-Ala-Asn(Trt)45-Lys (Boc) -Val-Gly-amino resin
In the preparation method of Bulevirtide, the amino resin has an amino substitution value of 0.2-0.8 mmol/g resin, and the preferable substitution value is 0.3-0.5 mmol/g resin.
In the preparation method of Bulevirtide, the amino resin is one of Rink MBHA resin, Rink Amide resin or Rink Amide AM resin, and Rink Amide MBHA resin is preferred.
In the Bulevirtide preparation method, the dosage of the Fmoc-protected amino acid or the protected amino acid fragment is 1.2-6 times of the total mole number of the charged resin; preferably 2.5 to 3.5 times.
In a preferred embodiment of the invention, the Bulevilide resin is subjected to acidolysis and resin and side chain protecting groups are removed simultaneously to obtain a Bulevilide element linear peptide crude product.
Further, an acidolysis agent adopted in acidolysis of the Bulevirtide resin is a mixed solvent of trifluoroacetic acid (TFA), 1, 2-Ethanedithiol (EDT) and water, and the mixture ratio of the mixed solvent is as follows: the TFA ratio is 80-95% (V/V), the EDT ratio is 1-10% (V/V), and the balance is water. The preferred formulation is 89-91% TFA, 4-6% EDT, and the balance water. Preferably, the mixture ratio is 90%, EDT 5% and the balance of water.
The dosage of the acidolysis agent is 4-15 ml of acidolysis agent per gram of Bulevilide resin, and preferably 9-11 ml of acidolysis agent per gram of Bulevilide resin. The time for cracking by using the acidolysis agent is 1-5 hours, preferably 2 hours at room temperature.
Further, the Bulevilide crude product is purified by high performance liquid chromatography and freeze-dried to obtain a Bulevilide pure product, and the specific method comprises the following steps:
adding water into the Bulevitide crude product, stirring, adjusting the pH value to 8.0 by using ammonia water until the Bulevitide crude product is completely dissolved, filtering the solution by using a 0.45-micrometer mixed microporous filter membrane, and purifying for later use;
purifying by high performance liquid chromatography, wherein the purification is carried out by using reversed phase C18 with chromatographic packing of 10 mu m, alternately purifying by using two mobile phase systems, the first mobile phase system is 0.1% TFA/water solution-0.1% TFA/acetonitrile solution, the second mobile phase system is 50mmol ammonium acetate/water solution-acetonitrile, the flow rate of a chromatographic column of 77mm 250mm is 90mL/min, eluting by using a gradient system, circularly injecting and purifying, sampling a crude product solution into the chromatographic column, starting mobile phase elution, collecting a main peak to evaporate acetonitrile, filtering by using a 0.45 mu m filter membrane to obtain a Bulevitide purified intermediate concentrated solution, adjusting the pH to 8.0, and preparing for desalting.
Desalting by high performance liquid chromatography with mobile phase system of water solution-acetonitrile, purification with reversed phase C18 with chromatographic packing of 10 μm, and chromatographic column flow rate of 77mm × 250mm of 90 mL/min; adopting a gradient elution and circulating sample loading method, loading a sample into a chromatographic column, starting mobile phase elution, collecting a spectrum, observing the change of the absorbance, collecting a main desalting peak, detecting the purity by using an analytical liquid phase, combining the main desalting peak solutions, concentrating under reduced pressure to obtain a Bulevirtide aqueous solution, and freeze-drying to obtain a Bulevirtide pure product.
The method of the invention directly uses the following special protected amino acids:
Myristony-Gly-Thr(tBu)-Asn(Trt)-Leu-Ser(tBu)-Val-Pro-Asn(Trt)-Pro-Leu-Gly-Phe-Phe-Pro-OH
shortens the production period, greatly improves the purity of the crude product, improves the product yield, and has wide practical value and application prospect.
Detailed Description
The invention discloses a method for synthesizing Bulevirtide, which can be realized by appropriately improving process parameters by referring to the content in the text. 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 of the present 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 of the methods described herein, as well as appropriate variations and combinations of the methods described herein, may be made and the techniques of the present invention employed without departing from the spirit and scope of the invention.
In the specific embodiment of the present invention, the Chinese meanings corresponding to the English abbreviations used in the application documents are shown in Table 1.
TABLE 1
English abbreviation | Name of Chinese | English abbreviation | Name of Chinese |
Fmoc | 9-fluorenylmethoxycarbonyl group | OtBu | Tert-butoxy radical |
tBu | Tert-butyl radical | Boc | Boc-acyl |
Trt | Trityl radical | Leu | Leucine |
Ser | Serine | Phe | Phenylalanine |
Glu | Glutamic acid | Thr | Threonine |
Trp | Tryptophan | Arg | Arginine |
Asp | Aspartic acid | Gln | Glutamine |
Ala | Alanine | Ile | Isoleucine |
Tyr | Tyrosine | His | Histidine |
Gly | Glycine | Lys | Lysine |
Val | Valine | Myristic acid | Myristic acid |
The invention is further illustrated by the following examples.
Example 1 Synthesis of Bulevirtide peptide resin by fragment ligation
Bulevilide peptide resin:
Myristony-Gly-Thr(tBu)-Asn(Trt)-Leu5-Ser(tBu)-Val-Pro-Asn(Trt)-Pro10-Leu-Gly-Phe-Phe-Pro15-Asp(OtBu)-His(Trt)-Gln(Trt)-Leu-Asp(OtBu)20-Pro-Ala-Phe-Gly-Ala25-Asn(Trt)-Ser(tBu)-Asn(Trt)-Asn(Trt)-Pro30-Asp(OtBu)-Trp(Boc)-Asp(OtBu)-Phe-Asn(Trt)35-Pro-Asn(Trt)-Lys(Boc)-Asp(OtBu)-His(Trt)40-Trp(Boc)-Pro-Glu(OtBu)-Ala-Asn(Trt)45-Lys (Boc) -Val-Gly-amino resin
Rink Amide MBHA resin is used as initial resin, and is sequentially coupled with protected amino acids shown in table 2 through Fmoc protection removal and coupling reaction to prepare Bulevitide peptide resin. The protection fragments used in this example were: Myristony-Gly-Thr (tBu) -Asn (Trt) -Leu-Ser (tBu) -Val-Pro-Asn (Trt) -Pro-Leu-Gly-Phe-Phe-Pro-OH
TABLE 2
1. Introduction of the 1 st protected amino acid
Dissolving 0.03mol of the 1 st protected amino acid and 0.03mol of HOBt in a proper amount of DMF; and adding 0.03mol DIC slowly into the protected amino acid DMF solution under stirring, and reacting for 30 minutes under stirring at room temperature to obtain an activated protected amino acid solution for later use.
0.01mol of Fmoc-Gly-resin (substitution value about 0.5mmol/g) was taken, deprotected with 20% PIP/DMF solution for 25 min, washed and filtered to give Fmoc-removed resin.
And adding the activated 1 st protected amino acid solution into the Fmoc-removed resin, performing coupling reaction for 120-300 minutes, and filtering and washing to obtain the resin containing 1 protected amino acid.
2. Inoculating 2-34 protected amino acids or fragments
And sequentially inoculating the corresponding 2 nd to 34 th protected amino acids or fragments by adopting the same method to obtain the Bulevirtide peptide resin.
Example 2 Synthesis of Bulevirtide peptide resin Using a conventional one-by-one grafting method
Bulevilide peptide resin:
Myristony-Gly-Thr(tBu)-Asn(Trt)-Leu5-Ser(tBu)-Val-Pro-Asn(Trt)-Pro10-Leu-Gly-Phe-Phe-Pro15-Asp(OtBu)-His(Trt)-Gln(Trt)-Leu-Asp(OtBu)20-Pro-Ala-Phe-Gly-Ala25-Asn(Trt)-Ser(tBu)-Asn(Trt)-Asn(Trt)-Pro30-Asp(OtBu)-Trp(Boc)-Asp(OtBu)-Phe-Asn(Trt)35-Pro-Asn(Trt)-Lys(Boc)-Asp(OtBu)-His(Trt)40-Trp(Boc)-Pro-Glu(OtBu)-Ala-Asn(Trt)45-Lys (Boc) -Val-Gly-amino resin
Rink Amide MBHA resin is used as initial resin, and is sequentially coupled with protected amino acids shown in table 2 through Fmoc protection removal and coupling reaction to prepare Bulevitide peptide resin.
TABLE 3
1. Introduction of the 1 st protected amino acid
Dissolving 0.03mol of the 1 st protected amino acid and 0.03mol of HOBt in a proper amount of DMF; and adding 0.03mol DIC slowly into the protected amino acid DMF solution under stirring, and reacting for 30 minutes under stirring at room temperature to obtain an activated protected amino acid solution for later use.
0.01mol of Fmoc-Gly-resin (substitution value about 0.5mmol/g) was taken, deprotected with 20% PIP/DMF solution for 25 min, washed and filtered to give Fmoc-removed resin.
And adding the activated 1 st protected amino acid solution into the Fmoc-removed resin, performing coupling reaction for 120-300 minutes, and filtering and washing to obtain the resin containing 1 protected amino acid.
2. Inoculating 2-48 protected amino acids or fatty acids
And sequentially inoculating the corresponding 2 nd to 48 th protected amino acids or fatty acids by adopting the same method to obtain the Bulevirtide peptide resin.
EXAMPLE 3 preparation of crude Bulevilide
Taking Bulevilide peptide resin prepared in example 1, adding a cleavage reagent (10 mL/g resin of the cleavage reagent) with the volume ratio of TFA, water and EDT (95: 5), uniformly stirring, stirring at room temperature for reaction for 3 hours, filtering a reaction mixture by using a sand core funnel, collecting a filtrate, washing the resin with a small amount of TFA for 3 times, combining the filtrates, concentrating under reduced pressure, adding anhydrous ether for precipitation, washing the precipitate with anhydrous ether for 3 times, and drying under reduced pressure at 35-45 ℃ to obtain a Bulevilide crude product with the purity of 67.3%.
EXAMPLE 4 preparation of crude Bulevilide
Taking Bulevilide peptide resin prepared in example 2, adding a cleavage reagent (10 mL/g resin of the cleavage reagent) with the volume ratio of TFA, water and EDT (95: 5), uniformly stirring, stirring at room temperature for reaction for 3 hours, filtering a reaction mixture by using a sand core funnel, collecting a filtrate, washing the resin with a small amount of TFA for 3 times, combining the filtrates, concentrating under reduced pressure, adding anhydrous ether for precipitation, washing the precipitate with anhydrous ether for 3 times, and drying under reduced pressure at 35-45 ℃ to obtain a Bulevilide crude product with the purity of 45.7%.
EXAMPLE 5 purification of crude Bulevilide
Taking the Bulevilide crude product prepared in the embodiment 3, adding water, stirring, adjusting the pH to 8.0 by using ammonia water until the Bulevilide crude product is completely dissolved, filtering the solution by using a 0.45-micron mixed microporous filter membrane, and purifying for later use;
purification was performed by high performance liquid chromatography using reverse phase C18 with 10 μm chromatography packing and alternating purification with two mobile phase systems, the first being 0.1% TFA/water-0.1% TFA/acetonitrile and the second being 50mmol ammonium acetate/water-acetonitrile. The flow rate of a chromatographic column of 77mm x 250mm is 90mL/min, a gradient system is adopted for elution, circulation sample injection purification is carried out, a crude product solution is taken to be loaded in the chromatographic column, mobile phase elution is started, a main peak is collected, acetonitrile is evaporated, and then a 0.45-micrometer filter membrane is used for filtration, so as to obtain a Bulevirtide purified intermediate concentrated solution, the acidity is adjusted to 8.0, and the Bulevirtide purified intermediate concentrated solution is prepared for desalination;
desalting by high performance liquid chromatography with mobile phase system of water solution-acetonitrile, purification with reversed phase C18 with chromatographic packing of 10 μm, and chromatographic column flow rate of 77mm × 250mm of 90 mL/min; the method comprises the steps of adopting a gradient elution and circulation loading method, loading a sample into a chromatographic column, starting mobile phase elution, collecting a map, observing the change of absorbance, collecting a desalted main peak, detecting the purity by using an analytical liquid phase, combining desalted main peak solutions, concentrating under reduced pressure to obtain a Bulevilide aqueous solution, and freeze-drying to obtain a Bulevilide pure product 13.7g, wherein the purity is 99.1%, the maximum single impurity content is 0.13%, the total yield is 25.4%, and the molecular weight is 5399.2 (100% M + H).
EXAMPLE 6 purification of crude Bulevilide
Taking the Bulevilide crude product prepared in the embodiment 4, adding water, stirring, adjusting the pH to 8.0 by using ammonia water until the Bulevilide crude product is completely dissolved, filtering the solution by using a 0.45-micron mixed microporous filter membrane, and purifying for later use;
purification was performed by high performance liquid chromatography using reverse phase C18 with 10 μm chromatography packing and alternating purification with two mobile phase systems, the first being 0.1% TFA/water-0.1% TFA/acetonitrile and the second being 50mmol ammonium acetate/water-acetonitrile. The flow rate of a chromatographic column of 77mm x 250mm is 90mL/min, a gradient system is adopted for elution, circulation sample injection purification is carried out, a crude product solution is taken to be loaded in the chromatographic column, mobile phase elution is started, a main peak is collected, acetonitrile is evaporated, and then a 0.45-micrometer filter membrane is used for filtration, so as to obtain a Bulevirtide purified intermediate concentrated solution, the acidity is adjusted to 8.0, and the Bulevirtide purified intermediate concentrated solution is prepared for desalination;
desalting by high performance liquid chromatography with mobile phase system of water solution-acetonitrile, purification with reversed phase C18 with chromatographic packing of 10 μm, and chromatographic column flow rate of 77mm × 250mm of 90 mL/min; the method comprises the steps of adopting a gradient elution and circulation loading method, loading a sample into a chromatographic column, starting mobile phase elution, collecting a map, observing the change of absorbance, collecting a desalted main peak, detecting the purity by using an analytical liquid phase, combining desalted main peak solutions, concentrating under reduced pressure to obtain a Bulevilide aqueous solution, and freeze-drying to obtain a Bulevilide pure product with the purity of 98.2 percent, the maximum single impurity content of 0.39 percent, the total yield of 12.8 percent and the molecular weight of 5399.0(100 percent M + H).
The above examples show that the purity of the product obtained by the method provided by the invention is more than 99.0%, and the total yield of the product is 25.4%, while the total yield of the conventional one-by-one access method is only 12.8%. The method obviously improves the purity of the crude product and the total yield of the product, and has wide practical value and application prospect.
Claims (6)
1. A method of preparing bulevitide, comprising: amino resin is adopted as initial resin, Bulevilide peptide resin is prepared by a solid-phase polypeptide synthesis method, Bulevilide peptide resin is subjected to acidolysis to obtain a Bulevilide crude product, and finally, the Bulevilide crude product is purified and freeze-dried to obtain a Bulevilide pure product:
Myristony-Gly-Thr-Asn-Leu5-Ser-Val-Pro-Asn-Pro10-Leu-Gly-
Phe-Phe-Pro15-Asp-His-Gln-Leu-Asp20-Pro-Ala-Phe-Gly-Ala25-
Asn-Ser-Asn-Asn-Pro30-Asp-Trp-Asp-Phe-Asn35-Pro-Asn-Lys-
Asp-His40-Trp-Pro-Glu-Ala-Asn45-Lys-Val-Gly-NH2
2. the method for producing Bulevilide as defined in claim 1, wherein: when Myristic acid at the 1 st position to Pro at the 15 th position are accessed together, the corresponding protected amino acid fragments are as follows:
Myristony-Gly-Thr(tBu)-Asn(Trt)-Leu-Ser(tBu)-Val-Pro-Asn(Trt)-Pro-Leu-Gly-Phe-
Phe-Pro-OH
3. the Bulevirtide preparation method according to claim 1, wherein the amino resin has an amino substitution value of 0.2 to 0.8mmol/g resin, preferably a substitution value of 0.3 to 0.5mmol/g resin.
4. The method for preparing Bulevilide as claimed in claim 1, wherein the amino resin is one of Rink MBHA resin, Rink Amide resin or Rink Amide AM resin, preferably Rink Amide MBHA resin.
5. The method for producing Bulevilide according to any one of claims 1 to 4, wherein: and carrying out acidolysis on the Bulevilide peptide resin, and simultaneously removing the resin and a side chain protecting group to obtain a Bulevilide crude product.
6. The method for producing Bulevilide as defined in claim 1, wherein: the crude Bulevilide product is purified by high performance liquid chromatography and freeze-dried to obtain a pure Bulevilide product.
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CN109354623A (en) * | 2012-04-25 | 2019-02-19 | 华辉安健(北京)生物科技有限公司 | The composition and related application of Hepaitis B virus functional receptor |
CN110818790A (en) * | 2019-10-31 | 2020-02-21 | 成都圣诺生物制药有限公司 | Preparation method of temeprelin |
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CN109354623A (en) * | 2012-04-25 | 2019-02-19 | 华辉安健(北京)生物科技有限公司 | The composition and related application of Hepaitis B virus functional receptor |
CN104781274A (en) * | 2012-11-12 | 2015-07-15 | 海德堡吕布莱希特-卡尔斯大学 | Lipopetides for use in treating liver diseases and cardiovascular diseases |
CN110818790A (en) * | 2019-10-31 | 2020-02-21 | 成都圣诺生物制药有限公司 | Preparation method of temeprelin |
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