CN106459164B - Snake venom C fragment polypeptide and analgesic drug - Google Patents

Snake venom C fragment polypeptide and analgesic drug Download PDF

Info

Publication number
CN106459164B
CN106459164B CN201580015678.XA CN201580015678A CN106459164B CN 106459164 B CN106459164 B CN 106459164B CN 201580015678 A CN201580015678 A CN 201580015678A CN 106459164 B CN106459164 B CN 106459164B
Authority
CN
China
Prior art keywords
residue
compound
acid
synthesis
snake venom
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Active
Application number
CN201580015678.XA
Other languages
Chinese (zh)
Other versions
CN106459164A (en
Inventor
孟庆雄
刘自成
支钦
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
SHENZHEN CITY JIANYUAN PHARMACEUTICAL TECHNOLOGY Co Ltd
Original Assignee
SHENZHEN CITY JIANYUAN PHARMACEUTICAL TECHNOLOGY Co Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by SHENZHEN CITY JIANYUAN PHARMACEUTICAL TECHNOLOGY Co Ltd filed Critical SHENZHEN CITY JIANYUAN PHARMACEUTICAL TECHNOLOGY Co Ltd
Publication of CN106459164A publication Critical patent/CN106459164A/en
Application granted granted Critical
Publication of CN106459164B publication Critical patent/CN106459164B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

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
    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
    • A61K38/08—Peptides having 5 to 11 amino acids
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
    • C07K7/04—Linear peptides containing only normal peptide links
    • C07K7/06—Linear peptides containing only normal peptide links having 5 to 11 amino acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Organic Chemistry (AREA)
  • Immunology (AREA)
  • Engineering & Computer Science (AREA)
  • Epidemiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Biochemistry (AREA)
  • Genetics & Genomics (AREA)
  • Molecular Biology (AREA)
  • Biophysics (AREA)
  • Toxicology (AREA)
  • Zoology (AREA)
  • Peptides Or Proteins (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

A snake venom C fragment polypeptide is provided, which is obtained by modifying a decapeptide derived from cobra neurotoxin-C. The polypeptide has analgesic effect, and also removes neurotoxicity of neurotoxin-C. Also provides an analgesic drug comprising the polypeptide.

Description

Snake venom C fragment polypeptide and analgesic drug
Technical Field
The invention relates to the field of polypeptide drugs, in particular to snake venom C fragment polypeptide with an analgesic effect and an analgesic drug.
Background
Pain is a common clinical symptom caused by various diseases, and the main purpose of the patients for asking medical questions is to relieve the pain. The analgesic drugs commonly used in clinic are mainly opioid potent analgesic drugs and antipyretic analgesic and anti-inflammatory drugs. The former is represented by morphine, which is a strong analgesic, and the latter is represented by aspirin, which is a weak analgesic. Opioid analgesics are narcotic analgesics, clinically limited to short-term analgesia of acute pain, such as intra-and post-surgical analgesia, bone fractures and acute visceral colic, and also used for intractable pain that some end-stage patients cannot treat, such as severe pain in advanced cancers. The medicine has the characteristics of quick response and strong analgesic effect. The disadvantages are addiction and quick tolerance (i.e. the curative effect is gradually reduced after a plurality of times of medication), and serious adverse reactions such as constipation, respiratory depression and the like. The antipyretic analgesic has the functions of resisting inflammation, allaying fever and relieving pain, and has wide clinical application. Can be used as analgesic for chronic pain. The medicine has the characteristics of mild analgesic effect and no addiction. The disadvantages are that the pain relieving effect is not strong enough, serious adverse reaction of digestive tract is caused, and individual medicine has cardiovascular side effect, liver toxicity or anaphylactic reaction. Chronic pain is one of the common clinical symptoms afflicting many patients, affecting quality of life and work, and the annual medical costs for chronic pain in the united states are as high as $ 260 billion, and analgesics that are powerful but not addictive and can be taken safely for long periods are currently lacking.
The cobra venom has been used for years at home and abroad to relieve malignant tumor pain, various neuralgia and arthralgia. The most obvious advantage of snake venom analgesia is that tolerance and addiction do not occur after continuous use, and side effects are little, and the analgesia is obviously different from morphine drugs. In China, Kunming animals separate neurotoxin-C from snake venom of Chinese cobra in the early years, and prepare analgesic Ketongning, and then develop compound Ketongning (containing Chinese cobra neurotoxin, tramadol and ibuprofen). The latter is currently clinically used for pain caused by cancer and the like with a good effect. At present, the clinical application is also cobratide injection prepared by Chinese cobra alpha-neotoxin and used for easing pain.
However, the traditional snake venom analgesic drugs have certain neurotoxicity, thereby causing certain adverse reactions and being not beneficial to clinical use.
Disclosure of Invention
Based on the above, there is a need for a snake venom C fragment polypeptide and an analgesic drug which have analgesic effects and are advantageous for clinical use.
A snake venom C fragment polypeptide having the following structural formula:
R1-Lys-Xaa1-His-Arg-Xaa2-Xaa3-Arg-Xaa4-Xaa5-Arg-R2；
wherein, -R1Has the structural formula of-NR3R4，-R3is-H, a carbon atomAlkyl groups in an amount of 1 to 16 or alkanoyl groups having 1 to 16 carbon atoms, -R4is-H, an alkyl group having 1 to 16 carbon atoms or an alkanoyl group having 1 to 16 carbon atoms;
-R2has a structural formula of-CONR5R6or-COOR7，-R5is-H or an alkanoyl group having 1 to 16 carbon atoms, -R6is-H or an alkanoyl group having 1 to 16 carbon atoms, -R7is-H or an alkanoyl group having 1 to 16 carbon atoms;
-Xaa 1-is an Asp residue or a Glu residue;
-Xaa 2-is a Gly residue, a Pro residue or a D-Pro residue;
-Xaa 3-is a Thr residue, a Ser residue, a D-Thr residue, a D-Ser residue or
Figure BDA0001118756140000021
-R is an alkyl group having 1 to 4 carbon atoms;
-Xaa 4-is an Ile residue, Leu residue, Nle residue, Val residue, Abu residue, Ala residue or Aib residue;
-Xaa 5-is an Asp residue or a Glu residue;
in one embodiment, -R1is-NH2。
In one embodiment, -R2is-COOH.
In one embodiment, -R3is-H, methyl, octyl, hexadecyl, acetyl, heptanoyl or palmitoyl, -R4is-H, methyl, octyl, hexadecyl, acetyl, heptanoyl or palmitoyl.
In one embodiment, -R5is-H, acetyl, heptanoyl or palmitoyl.
In one embodiment, -R6is-H, acetyl, heptanoyl or palmitoyl.
In one embodiment, -R7is-H, acetyl, heptanoyl or palmitoyl.
In one embodiment, -Xaa 3-is Thr (OCH)3) Residue, Ser (OCH)3) Residue or Ser (O)nCH3) And (c) a residue.
An analgesic drug comprises the snake venom C fragment polypeptide or a medically acceptable salt formed by the snake venom C fragment polypeptide and acid.
In one embodiment, the acid includes an organic acid and an inorganic acid, the inorganic acid is hydrochloric acid, sulfuric acid, or phosphoric acid, and the organic acid is acetic acid, oxalic acid, citric acid, fumaric acid, malic acid, or lactic acid.
In one embodiment, the solvent is water, ethanol or acetone.
The snake venom C fragment polypeptide is obtained by modifying a 10 peptide compound obtained by hydrolyzing, separating and purifying cobra neurotoxin-C, and tests prove that the snake venom C fragment polypeptide is an analgesic active center of the neurotoxin and has the characteristics of quick response, long action time, no tolerance, no addiction and the like. The snake venom C fragment polypeptide has good analgesic effect, and simultaneously removes neurotoxicity carried by original neurotoxin-C, so that the snake venom C fragment polypeptide greatly reduces the incidence rate of adverse reaction while maintaining the drug effect, and is beneficial to clinical use.
Detailed Description
The snake venom C fragment polypeptides and analgesic agents are described in further detail below with reference to specific examples.
One embodiment of a snake venom C fragment polypeptide has the following structural formula:
R1-Lys-Xaa1-His-Arg-Xaa2-Xaa3-Arg-Xaa4-Xaa5-Arg-R2。
R1-Lys-, -Xaa1-, -His-, -Arg-, -Xaa2-, -Xaa3, -Arg-, -Xaa4-, -Xaa 5-and-Arg-R2Are all amino acid residues.
-R1Has the structural formula of-NR3R4，-R3、-R4is-H, an alkyl group having 1 to 16 carbon atoms or an alkanoyl group having 1 to 16 carbon atoms.
In a preferred embodiment, -R3、-R4is-H, methyl, octyl, hexadecyl, acetyl, heptanoyl or palmitoyl.
In particular, -R1is-NH2。
-R2Has a structural formula of-CONR5R6or-COOR7，-R5、-R6、-R7is-H or an alkanoyl group having 1 to 16 carbon atoms.
In a preferred embodiment, -R5、-R6、-R7is-H, acetyl, heptanoyl or palmitoyl.
In particular, -R2is-COOH.
-Xaa 1-is an Asp residue or a Glu residue.
In a preferred embodiment, -Xaa 1-is a Glu residue.
-Xaa 2-is a Gly residue, a Pro residue or a D-Pro residue.
-Xaa 3-is a Thr residue, a Ser residue, a D-Thr residue, a D-Ser residue or
Figure BDA0001118756140000041
R is an alkyl group having 1 to 4 carbon atoms.
In a preferred embodiment, -Xaa 3-is Thr (OCH)3) Residue, Ser (OCH)3) Residue or Ser (O)nCH3) And (c) a residue.
In particular, -Xaa 3-is a Thr residue.
-Xaa 4-is an Ile residue, Leu residue, Nle residue, Val residue, Abu residue, Ala residue or Aib residue.
In a preferred embodiment, -Xaa 4-is an Ile residue.
-Xaa 5-is an Asp residue or a Glu residue.
In a preferred embodiment, -Xaa 5-is a Glu residue.
Specifically, the sequence of the snake venom C fragment polypeptide can be shown as SEQ ID No.1, SEQ ID No.2, SEQ ID No.3, SEQ ID No.4, SEQ ID No.5, SEQ ID No.6, SEQ ID No.7, SEQ ID No.8, SEQ ID No.9 or SEQ ID No. 10.
The snake venom C fragment polypeptide is obtained by modifying a 10 peptide compound obtained by hydrolyzing, separating and purifying cobra neurotoxin-C, and tests prove that the snake venom C fragment polypeptide is an analgesic active center of the neurotoxin and has the characteristics of quick response, long action time, no tolerance, no addiction and the like. The snake venom C fragment polypeptide has good analgesic effect, and simultaneously removes neurotoxicity carried by original neurotoxin-C, so that the snake venom C fragment polypeptide greatly reduces the incidence rate of adverse reaction while maintaining the drug effect, and is beneficial to clinical use.
The snake venom C fragment polypeptide can be used for relieving pain, in particular to relieving pain of cancer pain, postoperative pain, bone injury pain, arthritis pain, nerve pain and moderate and severe pain caused by other reasons.
The invention also discloses an analgesic drug, which comprises the snake venom C fragment polypeptide or a medically acceptable salt formed by the snake venom C fragment polypeptide and acid.
The acid includes organic acid and inorganic acid, the inorganic acid can be hydrochloric acid, sulfuric acid or phosphoric acid, and the organic acid can be acetic acid, oxalic acid, citric acid, fumaric acid, malic acid or lactic acid.
In other embodiments, the analgesic drug further comprises a solvent. The solvent may be water, ethanol or acetone.
The following are specific examples, and various instruments and reagents appearing in the examples are those which are conventional in the art, unless otherwise specified. The snake venom C fragment polypeptides are described in further detail below with reference to specific examples.
Example 1: preparation of compound 1.
In the compound (1), -R1is-NH2，-R2is-COOH, the sequence of the peptide chain is SEQ ID No.1
(1) Loading Wang resin (11mmol, substitution degree 1.0mmol/g) into a solid phase reaction column, and washing twice with DMF; adding DMF to swell for 30 min; dissolving Fmoc-Arg (Pbf) -OH, DMAP and HOBt in DMF, carrying out ice bath for 10 minutes, adding DIC, pre-activating for 2-5 min, adding the activated solution into a solid phase reaction column, stirring for reaction for 5h, draining, and washing with DMF for 6 times; washing with DCM for 3 times, shrinking MeOH for three times, and drying to obtain Fmoc-Arg (Pbf) -Wang resin with a detected substitution degree of 0.63 mmol/g; synthesis Scale 10.0 mmol.
(2) Adding DMF to completely immerse the resin in the solution, stirring for 30min, performing suction filtration on DMF, adding 20% DBLK for deprotection twice, performing suction drying, dissolving Fmoc-Glu (OtBu) -OH and HOBt in DMF, performing ice bath for 10 min, adding DIC, pre-activating for 2-5 min, adding the activated solution into a solid-phase reaction column, performing stirring reaction for 2h, and detecting the ninhydrin to be negative. Pumping, washing with DMF for 3 times, and deprotecting 20% DBLK twice and washing with DMF for 6 times; and (4) pumping out and coupling the next amino acid.
(3) Referring to the step (2), the remaining eight amino acids Fmoc-Ile-OH, Fmoc-Arg (Pbf) -OH, Fmoc-Thr (OtBu) -OH, Fmoc-Gly-OH, Fmoc-Arg (Pbf) -OH, Fmoc-His (Trt) -OH, Fmoc-Asp (OtBu) -OH and Fmoc-Lys (Pbf) -OH are condensed in sequence according to the peptide sequence, and then dried to obtain a resin peptide.
(4) Cracking: the resulting peptide resin was added in portions to the charge lysis solution (V) in ice bathTFA：VTIS：VH2O95: 2.5: 2.5), stirring for 30min, raising the temperature to room temperature, and stirring for 2.5 h. And (3) carrying out suction filtration on the lysate, washing the resin by using part of the lysate for two times, adding the washing liquid into the lysate, dropping the lysate into cold isopropyl ether, stirring vigorously, centrifuging, washing and drying to obtain crude peptide.
(5) And (3) carrying out gradient elution, purification and separation on the obtained crude product by adopting reverse-phase high performance liquid chromatography, wherein octadecylsilane chemically bonded silica is used as a stationary phase, a mobile phase A is 0.1% TFA water solution, and a mobile phase B is acetonitrile containing 0.1% TFA. The gradient program was: the initial state of the mobile phase B position 28%, keeping 10 minutes, then in 60 minutes the proportion of the mobile phase B is increased to 43%, the flow rate is 30mL/min, the detection wavelength is 214 nm. Collecting the fraction with the purity more than or equal to 98.0 percent and the single impurity less than or equal to 0.5 percent, concentrating, and freeze-drying to obtain a pure product. The molecular weight of the protamine was determined by LC-MS.
(6) The obtained product is subjected to salt conversion by 0.1 percent of HOAc/CH3CN to obtain an acetate product 1. HOAc.
(7) The product obtained was converted to the salt by 0.1% HCl/CH3CN to give the hydrochloride product 1. HCl.
Example 2: preparation of Compound (2).
In the compound (2), -R1is-N (Me)2，-R2is-COOH, and the sequence of the peptide chain is shown as SEQ ID No. 1.
Synthesis and purification were performed as described in example 1. Use of amino acid (Me) at site 10 in solid phase Synthesis2-Lys (Boc) -OH. The molecular weight of the protamine was determined by LC-MS.
Example 3: preparation of Compound (3).
In the compound (3), -R1is-N (octyl)2，-R2is-COOH, and the sequence of the peptide chain is shown as SEQ ID No. 1.
Synthesis and purification were performed as described in example 1. Use of an amino acid (Octyl) at position 10 in solid phase Synthesis2-Lys (Boc) -OH. The molecular weight of the protamine was determined by LC-MS.
Example 4: preparation of Compound (4).
In the compound (4), -R1is-N (Hexadecyl)2，-R2is-COOH, and the sequence of the peptide chain is shown as SEQ ID No. 1.
Synthesis and purification were performed as described in example 1. Use of amino acid at site 10 in solid phase synthesis (Hexadecyl)2-Lys (Boc) -OH. The molecular weight of the protamine was determined by LC-MS.
Example 5: preparation of Compound (5).
In the compound (5), -R1is-NH-Me, -R2is-COOH, and the sequence of the peptide chain is shown as SEQ ID No. 1.
Synthesis and purification were performed as described in example 1. The amino acids Fmoc-Me-Lys (Boc) -OH were used at site 10 in solid phase synthesis. The molecular weight of the protamine was determined by LC-MS.
Example 6: preparation of Compound (6).
In the compound (6), -R1is-NH-Octyl, -R2is-COOH, and the sequence of the peptide chain is shown as SEQ ID No. 1.
Synthesis and purification were performed as described in example 1. The amino acids Fmoc- (Octyl) -Lys (Boc) -OH were used at position 10 in solid phase synthesis. The molecular weight of the protamine was determined by LC-MS.
Example 7: preparation of Compound (7).
In the compound (7), -R1is-NH-Hexadecyl, -R2is-COOH, and the sequence of the peptide chain is shown as SEQ ID No. 1.
Synthesis and purification were performed as described in example 1. The amino acids Fmoc- (Hexadecyl) -Lys (Boc) -OH were used at site 10 in solid phase synthesis. The molecular weight of the protamine was determined by LC-MS.
Example 8: preparation of Compound (8).
In the compound (8), -R1is-NH2，-R2is-COO-Ac, and the sequence of the peptide chain is shown as SEQ ID No. 1.
Synthesis and purification were performed as described in example 1. The Resin carrier in solid phase synthesis is replaced by CTC Resin. After the coupling of the linear peptide was completed, it was cleaved with 20% TFE/DCM to give the product under the action of EtOH/DIEA/DMAP. The molecular weight of the protamine was determined by LC-MS.
Example 9: preparation of Compound (9).
In the compound (9), -R1is-NH2，-R2is-COO-octanyl, and the sequence of the peptide chain is shown as SEQ ID No. 1.
Synthesis and purification were performed as described in example 1. The Resin carrier in solid phase synthesis is replaced by CTC Resin. After coupling the linear peptide, cleavage with 20% TFE/DCM gave the product under octanol/DIC/DMAP. The molecular weight of the protamine was determined by LC-MS.
Example 10: preparation of Compound (10).
In the compound (10), -R1is-NH2，-R2is-COO-Pal, and the sequence of the peptide chain is shown as SEQ ID No. 1.
Synthesis and purification were performed as described in example 1. The Resin carrier in solid phase synthesis is replaced by CTC Resin. After coupling of the linear peptide, cleavage with 20% TFE/DCM gave the product as hexadecanol/DIC/DMAP. The molecular weight of the protamine was determined by LC-MS.
Example 11: preparation of Compound (11).
In the compound (11), -R1is-NH2，-R2is-CONH2The sequence of the peptide chain is shown as SEQ ID No. 1.
Synthesis and purification were performed as described in example 1. The resin carrier in solid phase synthesis is replaced by Fmoc Rink AmidemBHA. The molecular weight of the protamine was determined by LC-MS.
Example 12: preparation of Compound (12).
In the compound (12), -R1is-NH2，-R2is-CONH-Me, and the sequence of the peptide chain is shown in SEQ ID No. 1.
Synthesis and purification were performed as described in example 1. The Resin carrier in solid phase synthesis is replaced by CTC Resin. After the linear peptide is coupled, the linear peptide is cracked by using 20% TFE/DCM and reacted with methylamine hydrochloride under the action of HBTU/DIEA to obtain a product. The molecular weight of the protamine was determined by LC-MS.
Example 13: preparation of Compound (13).
In the compound (13), -R1is-NH2，-R2is-CONH-Octyl, and the sequence of the peptide chain is shown as SEQ ID No. 1.
Synthesis and purification were performed as described in example 1. The Resin carrier in solid phase synthesis is replaced by CTC Resin. After the linear peptide is coupled, the linear peptide is cracked by using 20% TFE/DCM and reacted with octylamine under the action of HBTU/DIEA to obtain a product. The molecular weight of the protamine was determined by LC-MS.
Example 14: preparation of Compound (14).
In the compound (14), -R1is-NH2，-R2is-CONH-Hexadecyl, and the sequence of the peptide chain is shown as SEQ ID No. 1.
Synthesis and purification were performed as described in example 1. The Resin carrier in solid phase synthesis is replaced by CTC Resin. After the coupling of the linear peptide is finished, the linear peptide is cracked by using 20% TFE/DCM and reacted with hexadecylamine under the action of HBTU/DIEA to obtain a product. The molecular weight of the protamine was determined by LC-MS.
Example 15: preparation of Compound (15).
In the compound (15), -R1is-NH2，-R2is-CON (Me)2And the sequence of the peptide chain is SEQ ID No.1, or a fragment thereof.
Synthesis and purification were performed as described in example 1. The Resin carrier in solid phase synthesis is replaced by CTC Resin. After the linear peptide is coupled, the linear peptide is cracked by using 20 percent TFE/DCM and reacted with dimethylamine hydrochloride under the action of HBTU/DIEA to obtain a product. The molecular weight of the protamine was determined by LC-MS.
Example 16: preparation of Compound (16).
In the compound (16), -R1is-NH2，-R2is-CON (octyl)2The sequence of the peptide chain is shown as SEQ ID No. 1.
Synthesis and purification were performed as described in example 1. The Resin carrier in solid phase synthesis is replaced by CTC Resin. After the linear peptide is coupled, the linear peptide is cracked by using 20% TFE/DCM and reacted with dioctylamine under the action of HBTU/DIEA to obtain a product. The molecular weight of the protamine was determined by LC-MS.
Example 17: preparation of Compound (17).
In the compound (17), -R1is-NH2，-R2is-CON (Hexadecyl)2The sequence of the peptide chain is shown as SEQ ID No. 1.
Synthesis and purification were performed as described in example 1. The Resin carrier in solid phase synthesis is replaced by CTC Resin. After the linear peptide was coupled, it was cleaved with 20% TFE/DCM and reacted with dicetylamine under HBTU/DIEA to give the product. The molecular weight of the protamine was determined by LC-MS.
Example 18: preparation of Compound (18).
In the compound (18), -R1is-NH2，-R2is-COOH, the sequence of the peptide chain is substantially identical to the sequence shown in SEQ ID No.1, differing only in that the amino acid at position Xaa1 is Glu.
Synthesis and purification were performed as described in example 1. The amino acids Fmoc-Glu (OtBu) -OH were used at position 9 in solid phase synthesis. The molecular weight of the protamine was determined by LC-MS.
Example 19: preparation of Compound (19).
In the compound (19), -R1is-NH2，-R2is-COOH, the sequence of the peptide chain is as shown in SEQ ID. 1 shows the sequence substantially identical, differing only in that the amino acid at position Xaa2 is Pro.
Synthesis and purification were performed as described in example 1. The amino acid Fmoc-Pro-OH was used at site 6 in solid phase synthesis. The molecular weight of the protamine was determined by LC-MS.
Example 20: preparation of Compound (20).
In the compound (20), -R1is-NH2，-R2is-COOH, the sequence of the peptide chain is substantially identical to the sequence shown in SEQ ID No.1, differing only in that the amino acid at position Xaa3 is Ser.
Synthesis and purification were performed as described in example 1. The amino acids Fmoc-Ser (OtBu) -OH were used at position 5 in solid phase synthesis. The molecular weight of the protamine was determined by LC-MS.
Example 21: preparation of Compound (21).
In the compound (21), -R1is-NH2，-R2is-COOH, the sequence of the peptide chain is substantially identical to the sequence shown in SEQ ID No.1, differing only in that the amino acid at position Xaa4 is Leu.
Synthesis and purification were performed as described in example 1. The amino acid Fmoc-Leu-OH was used at site 3 in solid phase synthesis. The molecular weight of the protamine was determined by LC-MS.
Example 22: preparation of Compound (22).
In the compound (22), -R1is-NH2，-R2is-COOH, the sequence of the peptide chain substantially corresponds to the sequence shown in SEQ ID No.1, with the only difference that the amino acid at position Xaa5 is Asp.
Synthesis and purification were performed as described in example 1. The amino acids Fmoc-Asp (OtBu) -OH were used in solid phase synthesis at position 2. The molecular weight of the protamine was determined by LC-MS.
Example 23: preparation of Compound (23).
In the compound (23), -R1is-NH2，-R2is-COOH, and the sequence of the peptide chain is the sequence shown in SEQ ID No. 2.
Synthesis and purification were performed as described in example 1. In the solid phase synthesis, the amino acids Fmoc-Asp (OtBu) -OH are used at the position 2, and the amino acids Fmoc-Glu (OtBu) -OH are used at the position 9. The molecular weight of the protamine was determined by LC-MS.
(6) The resulting product (23) was subjected to 0.1% HOAc/CH3Obtaining an acetate product 23. HOAc after CN is subjected to salt conversion
(7) The resulting product (23) was purified over 0.1% HCl/CH3CN is converted into hydrochloride product 23. HCl.
Example 24: preparation of Compound (24).
In the compound (24), -R1is-NH-Ac, -R2is-COOH, and the sequence of the peptide chain is the sequence shown in SEQ ID No. 2.
Synthesis and purification were performed as described in example 23. Use of Ac after amino acid coupling in solid phase Synthesis2O/DIEA end capping. The molecular weight of the protamine was determined by LC-MS.
Example 25: preparation of Compound (25).
In the compound (25), -R1is-NH-octanyl, -R2is-COOH, and the sequence of the peptide chain is the sequence shown in SEQ ID No. 2.
Synthesis and purification were performed as described in example 23. The capping with caprylic anhydride/DIEA was performed after the amino acid coupling in the solid phase synthesis. The molecular weight of the protamine was determined by LC-MS.
Example 26: preparation of Compound (26).
In the compound (26), -R1is-NH-Pal, -R2is-COOH, and the sequence of the peptide chain is the sequence shown in SEQ ID No. 2.
Synthesis and purification were performed as described in example 23. After the amino acid coupling in the solid phase synthesis was completed, the end capping was performed using palmitoyl chloride/DIEA. The molecular weight of the protamine was determined by LC-MS.
Example 27: preparation of Compound (27).
In the compound (27), -R1is-NH2，-R2is-COOH, the sequence of the peptide chain is substantially identical to the sequence shown in SEQ ID No.2, differing only in that the amino acid at position Xaa2 is Pro.
Synthesis and purification were performed as described in example 23. The amino acid Fmoc-Pro-OH was used at site 6 in solid phase synthesis. The molecular weight of the protamine was determined by LC-MS.
Example 28: preparation of Compound (28).
In the compound (28), -R1is-NH2，-R2is-COOH, the sequence of the peptide chain is substantially identical to the sequence shown in SEQ ID No.2, differing only in that the amino acid at position Xaa2 is D-Pro.
Synthesis and purification were performed as described in example 23. The amino acid Fmoc-D-Pro-OH was used at site 6 in solid phase synthesis. The molecular weight of the protamine was determined by LC-MS.
Example 29: preparation of Compound (29).
In the compound (29), -R1is-NH2，-R2is-COOH, the sequence of the peptide chain is substantially identical to the sequence shown in SEQ ID No.2, differing only in that the amino acid at position Xaa3 is Ser.
Synthesis and purification were performed as described in example 23. The amino acids Fmoc-Ser (OtBu) -OH were used at position 5 in solid phase synthesis. The molecular weight of the protamine was determined by LC-MS.
Example 30: preparation of Compound (30).
In the compound (30), -R1is-NH2，-R2is-COOH, the sequence of the peptide chain is substantially identical to the sequence shown in SEQ ID No.2, differing only in that the amino acid at position Xaa4 is Leu.
Synthesis and purification were performed as described in example 23. The amino acid Fmoc-Leu-OH was used at site 3 in solid phase synthesis. The molecular weight of the protamine was determined by LC-MS.
Example 31: preparation of Compound (31).
In the compound (31), -R1is-NH2，-R2is-COOH, the sequence of the peptide chain is substantially identical to the sequence shown in SEQ ID No.2, differing only in that the amino acid at position Xaa3 is D-Thr.
Synthesis and purification were performed as described in example 23. The amino acid Fmoc-D-Thr (OtBu) -OH was used at position 5 in solid phase synthesis. The molecular weight of the protamine was determined by LC-MS.
Example 32: preparation of Compound (32).
In the compound (32), -R1is-NH2，-R2is-COOH, the sequence of the peptide chain is substantially identical to the sequence shown in SEQ ID No.2, differing only in that the amino acid at position Xaa3 is D-Ser.
Synthesis and purification were performed as described in example 23. The amino acids Fmoc-D-Ser (OtBu) -OH were used at position 5 in solid phase synthesis. The molecular weight of the protamine was determined by LC-MS.
Example 33: preparation of Compound (33).
In the compound (33), -R1is-NH2，-R2is-COOH, the sequence of the peptide chain is substantially identical to the sequence shown in SEQ ID No.2, differing only in that the amino acid at position Xaa3 is Ser (OMe).
Synthesis and purification were performed as described in example 23. The amino acids Fmoc-Ser (OtBu) -OH were used at position 5 in solid phase synthesis. The molecular weight of the protamine was determined by LC-MS.
Example 34: preparation of Compound (34).
In the compound (34), -R1is-NH2，-R2is-COOH, the sequence of the peptide chain substantially corresponds to the sequence shown in SEQ ID No.2, differing only in that the amino acid at position Xaa3 is Ser (O)nBu)。
Synthesis and purification were performed as described in example 23. The amino acids Fmoc-Ser (OtBu) -OH were used at position 5 in solid phase synthesis. The molecular weight of the protamine was determined by LC-MS.
Example 35: preparation of Compound (35).
In the compound (35), -R1is-NH2，-R2is-COOH, the sequence of the peptide chain is substantially identical to the sequence shown in SEQ ID No.2, differing only in that the amino acid at position Xaa4 is Nle.
Synthesis and purification were performed as described in example 23. The amino acid Fmoc-Nle-OH was used at position 3 in solid phase synthesis. The molecular weight of the protamine was determined by LC-MS.
Example 36: preparation of Compound (36).
In the compound (36), -R1is-NH2，-R2Of the peptide chain-COOHThe sequence substantially corresponds to the sequence shown in SEQ ID No.2, and the only difference is that the amino acid at position Xaa4 is Val.
Synthesis and purification were performed as described in example 23. The amino acid Fmoc-Val-OH was used at site 3 in solid phase synthesis. The molecular weight of the protamine was determined by LC-MS.
Example 37: preparation of Compound (37).
In the compound (37), -R1is-NH2，-R2is-COOH, the sequence of the peptide chain is substantially identical to the sequence shown in SEQ ID No.2, differing only in that the amino acid at position Xaa4 is Abu.
Synthesis and purification were performed as described in example 23. The amino acid Fmoc-Abu-OH was used at position 3 in solid phase synthesis. The molecular weight of the protamine was determined by LC-MS.
Example 38: preparation of Compound (38).
In the compound (38), -R1is-NH2，-R2is-COOH, the sequence of the peptide chain substantially corresponds to the sequence shown in SEQ ID No.2, differing only in that the amino acid at position Xaa4 is Ala.
Synthesis and purification were performed as described in example 23. The amino acid Fmoc-Ala-OH was used at position 3 in the solid phase synthesis. The molecular weight of the protamine was determined by LC-MS.
Example 39: preparation of Compound (39).
In the compound (39), -R1is-NH2，-R2is-COOH, the sequence of the peptide chain is substantially identical to the sequence shown in SEQ ID No.2, differing only in that the amino acid at position Xaa4 is Aib.
Synthesis and purification were performed as described in example 23. The amino acid Fmoc-Aib-OH was used at position 3 in solid phase synthesis. The molecular weight of the protamine was determined by LC-MS.
Example 40: preparation of Compound (40).
In the compound (40), -R1is-NH2，-R2is-COOH, and the sequence of the peptide chain is shown as SEQ ID No. 3.
Synthesis and purification were performed as described in example 23. The molecular weight of the fine peptide was determined by LC-MS using the amino acids Fmoc-Glu (OtBu) -OH at position 2, Fmoc-Ser (OtBu) -OH at position 5 and Fmoc-Pro-OH at position 6 in solid phase synthesis.
Example 41: preparation of Compound (41).
In the compound (41), -R1is-NH2，-R2is-COOH, the sequence of the peptide chain is substantially identical to the sequence shown in SEQ ID No.3, differing only in that the amino acid at position Xaa3 is D-Thr.
Synthesis and purification were performed as described in example 23. The amino acid Fmoc-D-Thr (OtBu) -OH was used at position 5 in solid phase synthesis. The molecular weight of the protamine was determined by LC-MS.
Example 42: preparation of Compound (42).
In the compound (42), -R1is-NH2，-R2is-COOH, the sequence of the peptide chain is substantially identical to the sequence shown in SEQ ID No.3, differing only in that the amino acid at position Xaa2 is D-Pro.
Synthesis and purification were performed as described in example 23. The amino acid Fmoc-D-Pro-OH was used at site 6 in solid phase synthesis. The molecular weight of the protamine was determined by LC-MS.
Example 43: preparation of Compound (43).
In the compound (43), -R1is-NH2，-R2is-COOH, the sequence of the peptide chain is substantially identical to the sequence shown in SEQ ID No.3, differing only in that the amino acid at position Xaa3 is D-Ser.
Synthesis and purification were performed as described in example 23. The amino acids Fmoc-D-Ser (OtBu) -OH were used at position 5 in solid phase synthesis. The molecular weight of the protamine was determined by LC-MS.
Example 44: preparation of Compound (44).
In the compound (44), -R1is-NH-Pal, -R2is-COOH, and the sequence of the peptide chain is shown as SEQ ID No. 3.
Synthesis and purification were performed as described in example 23. After the amino acid coupling in the solid phase synthesis was completed, the end capping was performed using palmitoyl chloride/DIEA. The molecular weight of the protamine was determined by LC-MS.
Example 45: preparation of Compound (45).
In the compound (45), -R1is-NH2，-R2is-COOH, and the sequence of the peptide chain is shown as SEQ ID No. 4.
Synthesis and purification were performed as described in example 23. The amino acid Fmoc-Leu-OH was used at position 3 in solid phase synthesis, the amino acid Fmoc-Ser (OtBu) -OH was used at position 5 in solid phase synthesis, and the amino acid Fmoc-Asp (OtBu) -OH was used at position 9 in solid phase synthesis. The molecular weight of the protamine was determined by LC-MS.
Example 46: preparation of Compound (46).
In the compound (46), -R1is-NH-octanyl, -R2is-COOH, and the sequence of the peptide chain is shown as SEQ ID No. 4.
Synthesis and purification were performed as described in example 23. The capping with caprylic anhydride/DIEA was performed after the amino acid coupling in the solid phase synthesis. The molecular weight of the protamine was determined by LC-MS.
Example 47: preparation of Compound (47).
In the compound (47), -R1is-NH2，-R2is-COOH, and the sequence of the peptide chain is shown as SEQ ID No. 5.
Synthesis and purification were performed as described in example 23. The amino acids Fmoc-Glu (OtBu) -OH are used at position 2 in solid phase synthesis, the amino acids Fmoc-Leu-OH are used at position 3 in solid phase synthesis, and the amino acids Fmoc-Pro-OH are used at position 6 in solid phase synthesis. The molecular weight of the protamine was determined by LC-MS.
Example 48: preparation of Compound (48).
In the compound (48), -R1is-NH-Ac, -R2is-COOH, and the sequence of the peptide chain is shown as SEQ ID No. 5.
Synthesis and purification were performed as described in example 23. Use of Ac after amino acid coupling in solid phase Synthesis2O/DIEA end capping. The molecular weight of the protamine was determined by LC-MS.
Example 49: preparation of Compound (49).
In the compound (49), -R1is-NH2，-R2is-COOH, and the sequence of the peptide chain is shown as SEQ ID No. 6.
Synthesis and purification were performed as described in example 23. The amino acids Fmoc-Glu (OtBu) -OH are used at position 2 in solid phase synthesis, the amino acids Fmoc-Val-OH are used at position 3 in solid phase synthesis, and the amino acids Fmoc-Ser (OtBu) -OH are used at position 5 in solid phase synthesis. The molecular weight of the protamine was determined by LC-MS.
Example 50: preparation of Compound (50).
In the compound (50), -R1is-NH2，-R2is-CON (Me)2The sequence of the peptide chain is shown as SEQ ID No. 6.
Synthesis and purification were performed as described in example 23. The Resin carrier in solid phase synthesis is replaced by CTC Resin. After the linear peptide is coupled, the linear peptide is cracked by using 20 percent TFE/DCM and reacted with dimethylamine hydrochloride under the action of HBTU/DIEA to obtain a product. The molecular weight of the protamine was determined by LC-MS.
Example 51: preparation of Compound (51).
In the compound (51), -R1is-NH2，-R2is-COOH, and the sequence of the peptide chain is shown as SEQ ID No. 7.
Synthesis and purification were performed as described in example 1. The amino acid Fmoc-Leu-OH was used at position 3 in solid phase synthesis, the amino acid Fmoc-Ser (OtBu) -OH was used at position 5 in solid phase synthesis, and the amino acid Fmoc-Pro-OH was used at position 6 in solid phase synthesis. The molecular weight of the protamine was determined by LC-MS.
Example 52: preparation of Compound (52).
In the compound (52), -R1is-NH2，-R2is-CON (octyl)2The sequence of the peptide chain is shown as SEQ ID No. 7.
Synthesis and purification were performed as described in example 1. The Resin carrier in solid phase synthesis is replaced by CTC Resin. After the linear peptide is coupled, the linear peptide is cracked by using 20% TFE/DCM and reacted with dioctylamine under the action of HBTU/DIEA to obtain a product. The molecular weight of the protamine was determined by LC-MS.
Example 53: preparation of Compound (53).
In the compound (53), -R1is-NH2，-R2is-COOH, and the sequence of the peptide chain is shown as SEQ ID No. 8.
Synthesis and purification were performed as described in example 23. The amino acid Fmoc-Val-OH is used at position 3 in solid phase synthesis, the amino acid Fmoc-Ser (OtBu) -OH is used at position 5 in solid phase synthesis, and the amino acid Fmoc-Pro-OH is used at position 6 in solid phase synthesis. The molecular weight of the protamine was determined by LC-MS.
Example 54: preparation of Compound (54).
In the compound (54), -R1is-NH2，-R2is-CON (Hexadecyl)2The sequence of the peptide chain is shown as SEQ ID No. 8.
Synthesis and purification were performed as described in example 23. The Resin carrier in solid phase synthesis is replaced by CTC Resin. After the linear peptide was coupled, it was cleaved with 20% TFE/DCM and reacted with dicetylamine under HBTU/DIEA to give the product. The molecular weight of the protamine was determined by LC-MS.
Example 55: preparation of Compound (55).
In the compound (55), -R1is-NH2，-R2is-COOH, and the sequence of the peptide chain is shown as SEQ ID No. 9.
Synthesis and purification were performed as described in example 23. The amino acids Fmoc-Val-OH are used at position 3 in solid phase synthesis, the amino acids Fmoc-Ser (OtBu) -OH are used at position 5 in solid phase synthesis, the amino acids Fmoc-Pro-OH are used at position 6 in solid phase synthesis, and the amino acids Fmoc-Asp (OtBu) -OH are used at position 9 in synthesis. The molecular weight of the protamine was determined by LC-MS.
Example 56: preparation of Compound (56).
In the compound (56), -R1is-NH2，-R2is-CON (octyl)2The sequence of the peptide chain is shown as SEQ ID No. 9.
Synthesis and purification were performed as described in example 23. The Resin carrier in solid phase synthesis is replaced by CTC Resin. After the linear peptide is coupled, the linear peptide is cracked by using 20% TFE/DCM and reacted with dioctylamine under the action of HBTU/DIEA to obtain a product. The molecular weight of the protamine was determined by LC-MS.
Example 57: preparation of Compound (57).
In the compound (57), -R1is-NH2，-R2is-COOH, and the sequence of the peptide chain is shown as SEQ ID No. 10.
Synthesis and purification were performed as described in example 23. The amino acid Fmoc-Leu-OH was used at position 3 in solid phase synthesis, the amino acid Fmoc-Pro-OH was used at position 6 in solid phase synthesis, and the amino acid Fmoc-Asp (OtBu) -OH was used at position 9 in synthesis. The molecular weight of the protamine was determined by LC-MS.
Example 58: preparation of Compound (58).
In the compound (58), -R1is-NH2，-R2is-CON (octyl)2The sequence of the peptide chain is shown as SEQ ID No. 10.
Synthesis and purification were performed as described in example 23. The Resin carrier in solid phase synthesis is replaced by CTC Resin. After the linear peptide is coupled, the linear peptide is cracked by using 20% TFE/DCM and reacted with dioctylamine under the action of HBTU/DIEA to obtain a product. The molecular weight of the protamine was determined by LC-MS.
Example 59: analgesic test
The compounds (1) to (58) prepared in examples 1 to 58 were each prepared as a solution at a concentration of 0.5 mg/mL.
Mouse acetic acid writhing test: 590 male mice were selected and fasted for 10h before the experiment, and only water was supplied. The groups were randomized into 58 groups and a saline control group, 10 of which were administered. The compounds (1) to (58) are injected into the abdominal cavity of the mouse respectively, the analgesic agent is injected into the abdominal cavity of the mouse after 20min, 10mL/kg of 0.6 percent acetic acid solution is injected into the abdominal cavity, and the times of writhing of the mouse within 20min are recorded after 5 min.
The writhing inhibition ratio of each dose group was calculated according to the following formula to obtain the following table 1.
The writhing inhibition ratio is (number of writhing of control group-number of writhing of administration group)/number of writhing of control group x 100%
TABLE 1 Effect of the Compounds on writhing inhibition in mice (t/min)-1)
Figure BDA0001118756140000201
Figure BDA0001118756140000211
Figure BDA0001118756140000221
As can be seen from table 1, the compounds (1) to (58) prepared in examples 1 to 58 have significant inhibitory effects on mouse writhing, and are significantly different from the normal saline group, wherein the inhibitory rate of the compound (21) on mouse writhing is as high as 75.52%, and the analgesic effect is significant.
Figure IDA0001118756190000011
Figure IDA0001118756190000021
Figure IDA0001118756190000031
Figure IDA0001118756190000041
Figure IDA0001118756190000051

Claims (11)

1. A snake venom C fragment polypeptide having the structural formula:
R1-Lys-Xaa1-His-Arg-Xaa2-Xaa3-Arg-Xaa4-Xaa5-Arg-R2；
wherein, -R1Has the structural formula of-NR3R4，-R3is-H, an alkyl group having 1 to 16 carbon atoms or an alkanoyl group having 1 to 16 carbon atoms, -R4is-H,An alkyl group having 1 to 16 carbon atoms or an alkanoyl group having 1 to 16 carbon atoms;
-R2has a structural formula of-CONR5R6or-COOR7，-R5is-H or an alkanoyl group having 1 to 16 carbon atoms, -R6is-H or an alkanoyl group having 1 to 16 carbon atoms, -R7is-H or an alkanoyl group having 1 to 16 carbon atoms;
-Xaa 1-is an Asp residue or a Glu residue;
-Xaa 2-is a Gly residue, a Pro residue or a D-Pro residue;
-Xaa 3-is a Thr residue, a Ser residue, a D-Thr residue, a D-Ser residue or
Figure FDA0001118756130000011
-R is an alkyl group having 1 to 4 carbon atoms;
-Xaa 4-is an Ile residue, Leu residue, Nle residue, Val residue, Abu residue, Ala residue or Aib residue;
-Xaa 5-is an Asp residue or a Glu residue.
2. The snake venom C fragment polypeptide of claim 1, wherein-R1is-NH2。
3. The snake venom C fragment polypeptide of claim 1, wherein-R2is-COOH.
4. The snake venom C fragment polypeptide of claim 1, wherein-R3is-H, methyl, octyl, hexadecyl, acetyl, heptanoyl or palmitoyl, -R4is-H, methyl, octyl, hexadecyl, acetyl, heptanoyl or palmitoyl.
5. The snake venom C fragment polypeptide of claim 1, wherein-R5is-H, acetyl, heptanoyl or palmitoyl.
6. A snake according to claim 1Toxic C fragment polypeptide, characterized in that-R6is-H, acetyl, heptanoyl or palmitoyl.
7. The snake venom C fragment polypeptide of claim 1, wherein-R7is-H, acetyl, heptanoyl or palmitoyl.
8. The snake venom C fragment polypeptide of claim 1, wherein-Xaa 3-is Thr (OCH)3) Residue, Ser (OCH)3) Residue or Ser (O)nCH3) And (c) a residue.
9. An analgesic comprising a snake venom C fragment polypeptide of any one of claims 1-8 or a pharmaceutically acceptable salt thereof with an acid.
10. The analgesic drug of claim 9, wherein the acid comprises an organic acid and an inorganic acid, the inorganic acid is hydrochloric acid, sulfuric acid or phosphoric acid, and the organic acid is acetic acid, oxalic acid, citric acid, fumaric acid, malic acid or lactic acid.
11. The analgesic drug of claim 9, further comprising a solvent, wherein the solvent is water, ethanol or acetone.
CN201580015678.XA 2015-12-29 2015-12-29 Snake venom C fragment polypeptide and analgesic drug Active CN106459164B (en)

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/CN2015/099366 WO2017113086A1 (en) 2015-12-29 2015-12-29 Snake venom c polypeptide fragment and analgesic drug

Publications (2)

Publication Number Publication Date
CN106459164A CN106459164A (en) 2017-02-22
CN106459164B true CN106459164B (en) 2020-01-24

Family

ID=58094031

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201580015678.XA Active CN106459164B (en) 2015-12-29 2015-12-29 Snake venom C fragment polypeptide and analgesic drug

Country Status (2)

Country Link
CN (1) CN106459164B (en)
WO (1) WO2017113086A1 (en)

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1343689A (en) * 2000-09-18 2002-04-10 中山大学 Short-chain neurotoxin of sea serpent and gene for coding it

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1343689A (en) * 2000-09-18 2002-04-10 中山大学 Short-chain neurotoxin of sea serpent and gene for coding it

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
FRÉDÉRIC DUCANCEL等.NUCLEOTIDE SEQUENCE AND STRUCTURE ANALYSIS OF cDNAs ENCODING SHORT-CHAIN NEUROTOXINS FROM VENOM GLANDS OF A SEA SNAKE (AIPYSURUS LAEVIS).《Toxicon》.1990,119-123. *
immunization with a peptide having both T cell and conformationlly restricted B cell epitopes elicits neutralizing antisera against a snake neurotoxin;michel leonetti等;《The Journal of immunology》;19901215;4214-4221 *
Isolation, Properties and Amino Acid Sequences of Three Neurotoxins from the Venom of a Sea Snake, Aipysurus laevis;NOBUYO MAEDA等;《Biochem.J.》;19761231;79-87 *

Also Published As

Publication number Publication date
WO2017113086A1 (en) 2017-07-06
CN106459164A (en) 2017-02-22

Similar Documents

Publication Publication Date Title
US8629241B2 (en) Alpha helical mimics, their uses and methods for their production
CN102532301B (en) Novel Exendin-4 analogues and preparation method thereof
CN111410686A (en) Molecular modification of G L P-1R activator and application of dimer thereof in treating metabolic diseases
WO2008147556A2 (en) Melanocortin receptor ligands modified with hydantoin
TW201305197A (en) Glucose-dependent insulinotropic peptide analogs
KR20230088337A (en) GLP-1/GIP dual agonists
US20030158110A1 (en) Vasoactive intestinal peptide analogs
CN104098688A (en) Method for synthesizing thymalfasin
US20040122013A1 (en) Analogs of nocicettin
CN116554299A (en) Long-acting GLP-1 polypeptide analogue, and preparation method and application thereof
KR20230023658A (en) Long-acting GLP-1/GIP dual agonists
TW200819139A (en) Novel analogs of vasoactive intestinal peptide
CN106459164B (en) Snake venom C fragment polypeptide and analgesic drug
WO2022257831A1 (en) Polypeptide compounds using pd-1 receptors as targets, preparation method therefor and use thereof
EP1261626A1 (en) Bombesin analogs for treatment of cancer
CN111087474B (en) Hemp peptide (r) VD-Hp alpha and neuropeptide FF-based chimeric peptide and preparation method and application thereof
BG108587A (en) Peptides having antiangiogenic activity
CN108976287B (en) Disulfide bond cyclized analog of opioid and neuropeptide FF receptor multi-target molecule BN-9 and preparation method and application thereof
CN112961249B (en) Bifunctional peptide based on opioid peptide and cannabis peptide, and preparation method and application thereof
CN115073556A (en) Opioid/neuropeptide FF receptor multi-target cyclic peptide molecule and preparation and application thereof
CN109021117B (en) Opioid and neuropeptide FF receptor multi-target molecule BN-9 analogue, and preparation method and application thereof
CN107108706B (en) Snake venom C fragment polypeptide derivatives
CN112457369B (en) Preparation and application of long-acting analgesic peptide compounds acting on multiple targets
CN116514952B (en) GLP-1 analogues and application thereof
AU2005222732B2 (en) Alpha helical mimics, their uses and methods for their production

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant