CN117024564A - Polypeptide modified body and application thereof in preparation of anti-inflammatory and bactericidal medicines - Google Patents
Polypeptide modified body and application thereof in preparation of anti-inflammatory and bactericidal medicines Download PDFInfo
- Publication number
- CN117024564A CN117024564A CN202310942038.XA CN202310942038A CN117024564A CN 117024564 A CN117024564 A CN 117024564A CN 202310942038 A CN202310942038 A CN 202310942038A CN 117024564 A CN117024564 A CN 117024564A
- Authority
- CN
- China
- Prior art keywords
- polypeptide
- dmf
- amino acid
- gly
- lys
- 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.)
- Pending
Links
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 149
- 229920001184 polypeptide Polymers 0.000 title claims abstract description 147
- 102000004196 processed proteins & peptides Human genes 0.000 title claims abstract description 147
- 239000003814 drug Substances 0.000 title claims abstract description 17
- 230000000922 anti-bactericidal effect Effects 0.000 title claims abstract description 9
- 230000003110 anti-inflammatory effect Effects 0.000 title claims abstract description 9
- 238000002360 preparation method Methods 0.000 title claims description 4
- 229940079593 drug Drugs 0.000 title abstract description 13
- 125000003275 alpha amino acid group Chemical group 0.000 claims abstract 2
- 230000004048 modification Effects 0.000 claims description 25
- 238000012986 modification Methods 0.000 claims description 25
- 235000014113 dietary fatty acids Nutrition 0.000 claims description 8
- 229930195729 fatty acid Natural products 0.000 claims description 8
- 239000000194 fatty acid Substances 0.000 claims description 8
- 150000004665 fatty acids Chemical class 0.000 claims description 8
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 claims description 5
- 238000006482 condensation reaction Methods 0.000 claims description 5
- 230000018044 dehydration Effects 0.000 claims description 5
- 238000006297 dehydration reaction Methods 0.000 claims description 5
- POULHZVOKOAJMA-UHFFFAOYSA-N dodecanoic acid Chemical compound CCCCCCCCCCCC(O)=O POULHZVOKOAJMA-UHFFFAOYSA-N 0.000 claims description 4
- IPCSVZSSVZVIGE-UHFFFAOYSA-N hexadecanoic acid Chemical compound CCCCCCCCCCCCCCCC(O)=O IPCSVZSSVZVIGE-UHFFFAOYSA-N 0.000 claims description 4
- 125000001429 N-terminal alpha-amino-acid group Chemical group 0.000 claims description 3
- 239000005639 Lauric acid Substances 0.000 claims description 2
- 235000021314 Palmitic acid Nutrition 0.000 claims description 2
- WQEPLUUGTLDZJY-UHFFFAOYSA-N n-Pentadecanoic acid Natural products CCCCCCCCCCCCCCC(O)=O WQEPLUUGTLDZJY-UHFFFAOYSA-N 0.000 claims description 2
- TUNFSRHWOTWDNC-HKGQFRNVSA-N tetradecanoic acid Chemical compound CCCCCCCCCCCCC[14C](O)=O TUNFSRHWOTWDNC-HKGQFRNVSA-N 0.000 claims description 2
- 206010061126 Escherichia infection Diseases 0.000 claims 1
- 208000020612 escherichia coli infection Diseases 0.000 claims 1
- 206010040047 Sepsis Diseases 0.000 abstract description 32
- 230000004083 survival effect Effects 0.000 abstract description 29
- 230000001580 bacterial effect Effects 0.000 abstract description 10
- 210000004369 blood Anatomy 0.000 abstract description 6
- 239000008280 blood Substances 0.000 abstract description 6
- 230000003115 biocidal effect Effects 0.000 abstract description 5
- 208000035143 Bacterial infection Diseases 0.000 abstract description 4
- 208000022362 bacterial infectious disease Diseases 0.000 abstract description 4
- 238000002474 experimental method Methods 0.000 abstract description 4
- 210000000056 organ Anatomy 0.000 abstract description 4
- 206010059866 Drug resistance Diseases 0.000 abstract description 3
- 231100000135 cytotoxicity Toxicity 0.000 abstract description 3
- 230000003013 cytotoxicity Effects 0.000 abstract description 3
- 201000010099 disease Diseases 0.000 abstract description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract description 3
- 241000699670 Mus sp. Species 0.000 description 31
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 30
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 30
- 239000011347 resin Substances 0.000 description 25
- 229920005989 resin Polymers 0.000 description 25
- 239000000243 solution Substances 0.000 description 24
- 150000001413 amino acids Chemical group 0.000 description 22
- 238000004128 high performance liquid chromatography Methods 0.000 description 18
- 238000006243 chemical reaction Methods 0.000 description 15
- 238000005406 washing Methods 0.000 description 15
- 238000000746 purification Methods 0.000 description 14
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 10
- NQRYJNQNLNOLGT-UHFFFAOYSA-N Piperidine Chemical compound C1CCNCC1 NQRYJNQNLNOLGT-UHFFFAOYSA-N 0.000 description 10
- CMWYAOXYQATXSI-UHFFFAOYSA-N n,n-dimethylformamide;piperidine Chemical compound CN(C)C=O.C1CCNCC1 CMWYAOXYQATXSI-UHFFFAOYSA-N 0.000 description 10
- 239000002245 particle Substances 0.000 description 10
- 102400000888 Cholecystokinin-8 Human genes 0.000 description 9
- 101800005151 Cholecystokinin-8 Proteins 0.000 description 9
- 238000002784 cytotoxicity assay Methods 0.000 description 9
- 231100000263 cytotoxicity test Toxicity 0.000 description 9
- IZTQOLKUZKXIRV-YRVFCXMDSA-N sincalide Chemical compound C([C@@H](C(=O)N[C@@H](CCSC)C(=O)NCC(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(N)=O)NC(=O)[C@@H](N)CC(O)=O)C1=CC=C(OS(O)(=O)=O)C=C1 IZTQOLKUZKXIRV-YRVFCXMDSA-N 0.000 description 9
- 102000003688 G-Protein-Coupled Receptors Human genes 0.000 description 8
- 108090000045 G-Protein-Coupled Receptors Proteins 0.000 description 8
- 241000699666 Mus <mouse, genus> Species 0.000 description 8
- 238000012360 testing method Methods 0.000 description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 7
- 101100425538 Pseudomonas aeruginosa (strain UCBPP-PA14) tis1 gene Proteins 0.000 description 6
- 238000009833 condensation Methods 0.000 description 6
- 230000005494 condensation Effects 0.000 description 6
- 239000002173 cutting fluid Substances 0.000 description 6
- 238000005520 cutting process Methods 0.000 description 6
- 238000001514 detection method Methods 0.000 description 6
- 230000001376 precipitating effect Effects 0.000 description 6
- 125000006239 protecting group Chemical group 0.000 description 6
- 238000010189 synthetic method Methods 0.000 description 6
- JFLSOKIMYBSASW-UHFFFAOYSA-N 1-chloro-2-[chloro(diphenyl)methyl]benzene Chemical compound ClC1=CC=CC=C1C(Cl)(C=1C=CC=CC=1)C1=CC=CC=C1 JFLSOKIMYBSASW-UHFFFAOYSA-N 0.000 description 5
- TWJNQYPJQDRXPH-UHFFFAOYSA-N 2-cyanobenzohydrazide Chemical compound NNC(=O)C1=CC=CC=C1C#N TWJNQYPJQDRXPH-UHFFFAOYSA-N 0.000 description 5
- -1 Fmoc-Arg (pbf) -OH amino acid Chemical class 0.000 description 5
- 235000021360 Myristic acid Nutrition 0.000 description 5
- TUNFSRHWOTWDNC-UHFFFAOYSA-N Myristic acid Natural products CCCCCCCCCCCCCC(O)=O TUNFSRHWOTWDNC-UHFFFAOYSA-N 0.000 description 5
- JGFZNNIVVJXRND-UHFFFAOYSA-N N,N-Diisopropylethylamine (DIPEA) Chemical compound CCN(C(C)C)C(C)C JGFZNNIVVJXRND-UHFFFAOYSA-N 0.000 description 5
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 5
- 230000008859 change Effects 0.000 description 5
- 230000008878 coupling Effects 0.000 description 5
- 238000010168 coupling process Methods 0.000 description 5
- 238000005859 coupling reaction Methods 0.000 description 5
- 238000010511 deprotection reaction Methods 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 238000001914 filtration Methods 0.000 description 5
- 238000010438 heat treatment Methods 0.000 description 5
- 239000000203 mixture Substances 0.000 description 5
- FEMOMIGRRWSMCU-UHFFFAOYSA-N ninhydrin Chemical compound C1=CC=C2C(=O)C(O)(O)C(=O)C2=C1 FEMOMIGRRWSMCU-UHFFFAOYSA-N 0.000 description 5
- NNFCIKHAZHQZJG-UHFFFAOYSA-N potassium cyanide Chemical compound [K+].N#[C-] NNFCIKHAZHQZJG-UHFFFAOYSA-N 0.000 description 5
- 238000005086 pumping Methods 0.000 description 5
- 239000002904 solvent Substances 0.000 description 5
- 230000008961 swelling Effects 0.000 description 5
- 230000002924 anti-infective effect Effects 0.000 description 4
- 238000010828 elution Methods 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 3
- 238000013461 design Methods 0.000 description 3
- 238000004090 dissolution Methods 0.000 description 3
- 208000015181 infectious disease Diseases 0.000 description 3
- 238000001819 mass spectrum Methods 0.000 description 3
- 238000000329 molecular dynamics simulation Methods 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- CBPJQFCAFFNICX-IBGZPJMESA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-4-methylpentanoic acid Chemical compound C1=CC=C2C(COC(=O)N[C@@H](CC(C)C)C(O)=O)C3=CC=CC=C3C2=C1 CBPJQFCAFFNICX-IBGZPJMESA-N 0.000 description 2
- UMRUUWFGLGNQLI-QFIPXVFZSA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-6-[(2-methylpropan-2-yl)oxycarbonylamino]hexanoic acid Chemical compound C1=CC=C2C(COC(=O)N[C@@H](CCCCNC(=O)OC(C)(C)C)C(O)=O)C3=CC=CC=C3C2=C1 UMRUUWFGLGNQLI-QFIPXVFZSA-N 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 2
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- 239000003242 anti bacterial agent Substances 0.000 description 2
- 230000000844 anti-bacterial effect Effects 0.000 description 2
- 229940088710 antibiotic agent Drugs 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 210000004979 bone marrow derived macrophage Anatomy 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 239000007928 intraperitoneal injection Substances 0.000 description 2
- 210000003734 kidney Anatomy 0.000 description 2
- 210000004185 liver Anatomy 0.000 description 2
- 210000004072 lung Anatomy 0.000 description 2
- 230000014759 maintenance of location Effects 0.000 description 2
- 238000000034 method Methods 0.000 description 2
- 239000002504 physiological saline solution Substances 0.000 description 2
- 230000001681 protective effect Effects 0.000 description 2
- 238000012827 research and development Methods 0.000 description 2
- 210000000952 spleen Anatomy 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 230000001225 therapeutic effect Effects 0.000 description 2
- 238000002560 therapeutic procedure Methods 0.000 description 2
- IXAYZHCPEYTWHW-IBGZPJMESA-N (2r)-3-tert-butylsulfanyl-2-(9h-fluoren-9-ylmethoxycarbonylamino)propanoic acid Chemical compound C1=CC=C2C(COC(=O)N[C@@H](CSC(C)(C)C)C(O)=O)C3=CC=CC=C3C2=C1 IXAYZHCPEYTWHW-IBGZPJMESA-N 0.000 description 1
- 241000588626 Acinetobacter baumannii Species 0.000 description 1
- 108090000204 Dipeptidase 1 Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 241000588747 Klebsiella pneumoniae Species 0.000 description 1
- 102000018697 Membrane Proteins Human genes 0.000 description 1
- 108010052285 Membrane Proteins Proteins 0.000 description 1
- 125000000729 N-terminal amino-acid group Chemical group 0.000 description 1
- 241000589517 Pseudomonas aeruginosa Species 0.000 description 1
- PMZXXNPJQYDFJX-UHFFFAOYSA-N acetonitrile;2,2,2-trifluoroacetic acid Chemical compound CC#N.OC(=O)C(F)(F)F PMZXXNPJQYDFJX-UHFFFAOYSA-N 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 229940124425 anti-infective immunomodulator Drugs 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 102000006635 beta-lactamase Human genes 0.000 description 1
- 229940041011 carbapenems Drugs 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 230000003833 cell viability Effects 0.000 description 1
- 125000005313 fatty acid group Chemical group 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 230000002519 immonomodulatory effect Effects 0.000 description 1
- 230000003832 immune regulation Effects 0.000 description 1
- 229940124622 immune-modulator drug Drugs 0.000 description 1
- 239000002955 immunomodulating agent Substances 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 238000004811 liquid chromatography Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 231100001083 no cytotoxicity Toxicity 0.000 description 1
- 230000035778 pathophysiological process Effects 0.000 description 1
- 230000035515 penetration Effects 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 102000005962 receptors Human genes 0.000 description 1
- 108020003175 receptors Proteins 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
Classifications
-
- 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
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
-
- 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
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Oncology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biochemistry (AREA)
- Communicable Diseases (AREA)
- Biophysics (AREA)
- Genetics & Genomics (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention discloses a polypeptide modified body and application thereof in preparing anti-inflammatory and bactericidal medicines. The amino acid sequence of the polypeptide is shown as follows: gly (Gly) 1 ‑Val 2 ‑Lys 3 ‑Leu 4 ‑Tyr 5 ‑Gly 6 ‑Ser 7 ‑Asp 8 ‑Lys 9 ‑Ser 10 . Experiments show that the polypeptide modified body is basically free of cytotoxicity, can effectively reduce bacterial loads of blood and organs of a sepsis mouse, and can obviously improve the survival rate of the sepsis mouse; the polypeptide modified body can be used for treating diseases caused by bacterial infection, and provides a new solution for the current antibiotic drug resistance problem.
Description
The invention is as follows: 202111359409.9, filing date: division of the chinese invention patent application at 2021, 11, 17.
Technical Field
The invention belongs to the technical field of biological medicines, and particularly relates to a polypeptide modified body and application thereof in preparing anti-inflammatory and bactericidal medicines.
Background
At present, the clinically common drug-resistant bacteria such as Klebsiella pneumoniae, pseudomonas aeruginosa, acinetobacter baumannii and the like which produce ultra-broad-spectrum beta-lactamase escherichia coli, resist carbapenems cause the treatment of severe infection to become endless, and greatly increase the death rate of patients; antibiotic resistance has become a major medical problem that needs to be addressed in human health.
The current approaches for solving the clinical problem of antibiotic resistance mainly comprise: (1) developing novel antibiotics; (2) development of anti-infective immunomodulating agents. Because the research and development of the current novel antibiotics is in a bottleneck period, a significant breakthrough cannot be obtained; therefore, anti-infective immunomodulators are becoming the mainstream of research and development.
The G protein coupled receptors are membrane protein receptors with seven transmembrane helices and participate in most of vital activities in organisms, so that each G protein coupled receptor can become a potential therapeutic target; of the drugs currently marketed, about 20% belong to the class of GPCR-related drugs. G protein coupled receptor is abundantly expressed on various cell membranes of organisms, and plays an important role in various pathophysiological processes of organisms, in particular in anti-infective immune regulation.
The development of anti-infective immunomodulating drugs based on G protein coupled receptors can provide a new idea for clinical anti-infective immunomodulating treatment.
Disclosure of Invention
The invention aims to provide a polypeptide modified body and application thereof in preparing anti-inflammatory and bactericidal medicines.
In order to achieve the above object, the present invention has the following technical scheme:
a polypeptide having an amino acid sequence as set forth in SEQ ID No. 1.
The invention identifies and obtains the polypeptide (named RH-000) from the full-length structure of the G protein coupled receptor, and the amino acid sequence is as follows:
Ile 1 -Glu 2 -Arg 3 -His 4 -Val 5 -Ala 6 -Ile 7 -Ala 8 -Lys 9 -Val 10 -Lys 11 -Leu 12 -Tyr 13 -Gly 14 -Ser 15 -Asp 16 -Lys 17 -Ser 18 -Cys 19 -Arg 20 ;
the research shows that the polypeptide can obviously improve the survival rate of a sepsis mouse, and the survival rate of the sepsis mouse reaches more than 50 percent in 72 hours, which indicates that the polypeptide can be used for treating diseases caused by bacterial infection and provides a new solution for the current antibiotic drug resistance problem.
Therefore, the invention also provides application of the polypeptide in preparing anti-inflammatory and bactericidal medicines.
According to the invention, the active region of the polypeptide is analyzed through protein design and molecular dynamics simulation, and the structure of the polypeptide is modified based on the active epitope, so that a series of polypeptide modified bodies are obtained.
Therefore, the invention also provides a polypeptide modified body, which is obtained by modifying the polypeptide.
The polypeptide modified body has the amino acid sequence shown as follows:
Gly 1 -Val 2 -Lys 3 -Leu 4 -Tyr 5 -Gly 6 -Ser 7 -Asp 8 -Lys 9 -Ser 10 named RH-001;
or Gly 1 -Val 2 -Lys 3 -Leu 4 -Tyr 5 Named RH-002;
or Gly 1 -Val 2 -Ala 3 -Ile 4 -Ala 5 -Lys 6 -Val 7 -Lys 8 -Leu 9 -Tyr 10 Named RH-003;
or Gly 1 -Ile 2 -Glu 3 -Arg 4 -His 5 -Val 6 -Ala 7 -Ile 8 -Ala 9 -Lys 10 -Val 11 Named RH-004.
Experiments show that the polypeptide modified body is basically free of cytotoxicity, can effectively reduce bacterial loads of blood and organs of a sepsis mouse, and can obviously improve the survival rate of the sepsis mouse; the survival rate of 72h of the sepsis mice in RH-001 and RH-002 treatment groups is 89% and 67% respectively, which is obviously better than that of the RH-000 treatment group with the 72h survival rate of 56% of the sepsis mice, and the RH-003 can improve the 72h survival rate of the sepsis mice to 100%, so that the bacterial infection resistance is more remarkable.
Preferably, in the polypeptide modified body, a fatty acid is linked to the free carboxyl group of the N-terminal amino acid by a dehydration condensation reaction. The fatty acid group is added at the N end, so that on one hand, the permeability of the polypeptide can be enhanced, and on the other hand, the stability of the polypeptide can be improved, and the polypeptide can exert due effects.
More preferably, the free carboxyl group at the N-terminal Gly of the polypeptide modified RH-004 is linked with a fatty acid by dehydration condensation reaction. Treatment of sepsis mice with the polypeptide modification RH-M-004 obtained by further modification revealed that the 72h survival rate of sepsis mice also reached 100%.
Preferably, in the polypeptide modified body, the fatty acid is myristic acid, lauric acid or palmitic acid.
Based on the excellent performance of the polypeptide modified body, the invention also provides application of the polypeptide modified body in preparing anti-inflammatory and bactericidal medicines.
Compared with the prior art, the invention has the beneficial effects that:
(1) According to the invention, the polypeptide RH-000 is identified and obtained from the full-length structure of the G protein coupled receptor, and researches show that the polypeptide can obviously improve the survival rate of sepsis mice, and the survival rate of the sepsis mice reaches more than 50% in 72 hours, so that the polypeptide can be used for treating diseases caused by bacterial infection, and a new solution is provided for the current antibiotic drug resistance problem.
(2) According to the invention, the active region of the polypeptide is analyzed through protein design and molecular dynamics simulation, and the structure of the polypeptide is modified based on an active epitope, so that a series of polypeptide modified bodies RH-001, RH-002, RH-003 and RH-004 are obtained; experiments show that the polypeptide modified body is basically free of cytotoxicity, can effectively reduce bacterial loads of blood and organs of a sepsis mouse, and can obviously improve the survival rate of the sepsis mouse; the survival rate of 72h of the sepsis mice in RH-001 and RH-002 treatment groups is 89% and 67% respectively, which is obviously superior to that of the RH-000 treatment group with the 72h survival rate of 56% of that of the sepsis mice, and the RH-003 can improve the 72h survival rate of the sepsis mice to 100%, so that the sepsis mice have more remarkable antibacterial infection resistance and larger application potential.
(3) According to the invention, fatty acid is further connected to the free carboxyl of the N-terminal amino acid of each polypeptide modification through dehydration condensation reaction, and researches show that the polypeptide modification RH-004 can improve the 72h survival rate of sepsis mice to 100% after being connected with the fatty acid, so that the antibacterial infection resistance is optimized in one step, and the application potential is greater.
Drawings
FIG. 1 is a graph showing the results of HPLC purification of polypeptide RH-000 of the present invention;
wherein, pump A:0.065%trifluoroacetic in 100%water (v/v) represents mobile phase A: an aqueous trifluoroacetic acid solution having a volume fraction of 0.065%; pump B:0.05%trifluoroacetic in 100%acetonitrile (v/v) represents mobile phase B: acetonitrile trifluoroacetate solution with volume fraction of 0.05%; total Flow:1ml/min represents the flow rate: 1 ml/min; wavelength:220nm represents a wavelength:220 nm; time represents gradient elution Time, module represents elution program, command represents elution instruction, value represents elution volume; column Performance Column performance, detector a, column: inertsil OD-3.6X105 mm represents (chromatographic) column: (model number) Inertsil OD-3, (specification number) 4.6X250 mm; chromatogram represents a Chromatogram; mV represents the response intensity, min represents (retention time)/min; detecter A Channel 1 and 220nm indicate that the detection wavelength of the detector A is 220nm; peak Table shows Peak Table, peak shows Peak, ret. Time shows retention time, area shows Peak Area, height shows Peak Height, total shows sum; the following is the same;
FIG. 2 is a graph showing the mass spectrum identification result of the polypeptide RH-000 of the present invention;
wherein Mass Spectrum represents Mass Spectrum; positve Intensity positive intensity, m/z mass to charge ratio, the same applies below;
FIG. 3 is a graph showing the MASS identification of the polypeptide RH-001 of the present invention;
FIG. 4 is a graph showing the results of HPLC purification of polypeptide RH-001 of the present invention;
wherein LC Time Program represents a liquid chromatography gradient elution procedure; equipment is: ZJ20010140 represents a device (model number): ZJ20010140; the following is the same;
FIG. 5 is a graph showing the results of HPLC purification of polypeptide RH-002 of the present invention;
FIG. 6 is a graph showing the MASS identification of the polypeptide RH-002 of the present invention;
FIG. 7 is a graph showing MASS identification of polypeptide RH-003 of the present invention;
FIG. 8 is a graph showing the results of HPLC purification of polypeptide RH-003 of the present invention;
FIG. 9 is a graph showing the results of HPLC purification of polypeptide RH-004 of the present invention;
FIG. 10 is a graph showing the MASS identification of polypeptide RH-004 of the present invention;
FIG. 11 is a graph showing the MASS identification of the polypeptide RH-M-001 of the present invention;
FIG. 12 is a graph showing the results of HPLC purification of polypeptide RH-M-001 of the present invention;
FIG. 13 is a graph showing HPLC purification results of polypeptide RH-M-002 of the present invention;
FIG. 14 is a graph showing the MASS identification of the polypeptide RH-M-002 of the present invention;
FIG. 15 is a graph showing the MASS identification of the polypeptide RH-M-003 of the present invention;
FIG. 16 is a graph showing HPLC purification results for polypeptide RH-M-003 of the present invention;
FIG. 17 is a graph showing the results of HPLC purification of polypeptide RH-M-004 of the present invention;
FIG. 18 is a graph showing the MASS identification of polypeptide RH-M-004 of the present invention;
FIG. 19 shows the results of a CCK8 cytotoxicity assay of the polypeptide variant RH-001 of the present invention;
FIG. 20 shows the results of a CCK8 cytotoxicity assay of the polypeptide variant RH-002 of the present invention;
FIG. 21 shows the results of a CCK8 cytotoxicity assay of the polypeptide variant RH-003 of the present invention;
FIG. 22 shows the results of a CCK8 cytotoxicity assay of the polypeptide variant RH-004 of the present invention;
FIG. 23 shows the results of a CCK8 cytotoxicity assay of the polypeptide variant RH-M-001 of the present invention;
FIG. 24 shows the results of a CCK8 cytotoxicity assay of the polypeptide variant RH-M-002 of the present invention;
FIG. 25 shows the results of a CCK8 cytotoxicity assay of the polypeptide variant RH-M-003 of the present invention;
FIG. 26 shows the results of a CCK8 cytotoxicity assay of the polypeptide variant RH-M-004 of the present invention;
FIG. 27 is a Kaplan-Meier survival curve of sepsis mice treated with polypeptide RH-000, polypeptide modifications RH-001, RH-002, RH-003 and RH-004 of the present invention;
FIG. 28 is a Kaplan-Meier survival curve of sepsis mice treated with polypeptide RH-000, polypeptide modifications RH-M-001, RH-M-002, RH-M-003, and RH-M-004 of the present invention;
FIG. 29 shows the change in bacterial load in blood of sepsis mice following treatment with polypeptide modifications RH-001, RH-002, RH-003 and RH-M-004;
FIG. 30 shows the change in bacterial load in the liver of sepsis mice following treatment with polypeptide modifications RH-001, RH-002, RH-003 and RH-M-004;
FIG. 31 is a graph showing the change in bacterial load in spleen of sepsis mice after treatment with polypeptide modifications RH-001, RH-002, RH-003 and RH-M-004;
FIG. 32 shows the change in bacterial load in the lungs of sepsis mice following treatment with polypeptide modifications RH-001, RH-002, RH-003 and RH-M-004;
FIG. 33 shows the change in bacterial load in the kidneys of sepsis mice following treatment with polypeptide modifications RH-001, RH-002, RH-003 and RH-M-004.
Detailed Description
The technical scheme of the invention is further described in detail below with reference to the attached drawings and the detailed description.
Example 1
In this example, polypeptide RH-000 is identified and obtained from the full-length structure of the G protein coupled receptor, and the amino acid sequence of the polypeptide RH-000 is shown in SEQ ID No. 1:
Ile 1 -Glu 2 -Arg 3 -His 4 -Val 5 -Ala 6 -Ile 7 -Ala 8 -Lys 9 -Val 10 -Lys 11 -Leu 12 -Tyr 13 -Gly 14 -Ser 15 -Asp 16 -Lys 17 -Ser 18 -Cys 19 -Arg 20 ;
the synthetic method of the polypeptide RH-000 comprises the following steps:
(1) Swelling of the resin: placing 2-Chlorotrityl Chloride Resin resin into a reaction tube, adding DMF (15 ml/g), and oscillating for 30min;
(2) Coupling a first amino acid: filtering out the solvent through a sand core, taking Fmoc-Arg (pbf) -OH amino acid which is equal to the resin in mole, adding DIEA with 10 times of mole excess and HBTU with three times of mole excess, finally adding DMF for dissolution, and oscillating for 30min;
(3) Deprotection: DMF was removed, 20% piperidine DMF solution (15 ml/g) was added for 5min, and 20% piperidine DMF solution (15 ml/g) was removed for 15min;
(4) And (3) detection: pumping off piperidine solution, taking more than ten resin particles, washing with ethanol for three times, adding ninhydrin, KCN and phenol solution into the resin particles, heating the mixture at 105-110 ℃ for 5min to turn deep blue into positive reaction;
(5) Washing: DMF (10 ml/g) was twice, methanol (10 ml/g) was twice, DMF (10 ml/g) was twice;
(6) Condensation: triple excess of protected amino acid (Fmoc-Cys (tBu) -OH), triple excess of HBTU, all dissolved with as little DMF as possible, added to the reaction tube, immediately added with ten times excess NMM for 30min;
(7) Washing: DMF (10 ml/g) once, methanol (10 ml/g) twice, DMF (10 ml/g) twice;
(8) Repeating the steps (2) to (6), and connecting the amino acids in the sequences from right to left;
(9) Cleaving the polypeptide side chain protecting group, and then precipitating the polypeptide with cold diethyl ether;
preparing cutting fluid (10/g): TFA94.5%; 2.5% of water; EDT 2.5%; TIS1%;
cutting time: 120min;
(10) The ether precipitated polypeptide product was dried and then identified, and purified by HPLC (FIG. 1) and Mass identified (FIG. 2) to obtain the polypeptide RH-000 of this example.
Example 2
In this example, the structure of polypeptide RH-000 was further modified to obtain a series of polypeptide modifications, each having the amino acid sequence shown in Table 1.
Table 1 polypeptide variants and amino acid sequences thereof
As can be seen from Table 1, in this example, the region of action of polypeptide RH-000 was first analyzed by protein design and molecular dynamics simulation, and the structure of polypeptide RH-000 was modified based on the action epitope, so that polypeptide modified RH-001, RH-002, RH-003 and RH-004 were obtained; and then, further carrying out dehydration condensation reaction on the hydroxyl of fatty acid and free carboxyl on N-terminal amino acid residues, and modifying myristic acid at N-terminal of polypeptide modified bodies RH-001, RH-002, RH-003 and RH-004 to further obtain polypeptide modified bodies RH-M-001, RH-M-002, RH-M-003 and RH-M-004.
Wherein, the synthetic method of the polypeptide remodelling body RH-001 comprises the following steps:
(1) Swelling of the resin: placing 2-Chlorotrityl Chloride Resin resin into a reaction tube, adding DMF (15 ml/g), and oscillating for 30min;
(2) Coupling a first amino acid: filtering out the solvent by sand core suction, taking Fmoc-Ser-OH amino acid with the same mole as resin, adding 10 times of excessive DIEA, three times of HBTU, adding DMF to dissolve, and oscillating for 30min;
(3) Deprotection: DMF was removed, 20% piperidine DMF solution (15 ml/g) was added for 5min, and 20% piperidine DMF solution (15 ml/g) was removed for 15min;
(4) And (3) detection: pumping off piperidine solution, taking more than ten resin particles, washing with ethanol for three times, adding ninhydrin, KCN and phenol solution into the resin particles, heating the mixture at 105-110 ℃ for 5min to turn deep blue into positive reaction;
(5) Washing: DMF (10 ml/g) was twice, methanol (10 ml/g) was twice, DMF (10 ml/g) was twice;
(6) Condensation: triple excess of protected amino acid (Fmoc-Lys (Boc) -OH), triple excess of HBTU, all dissolved with as little DMF as possible, added to the reaction tube, immediately added to NMM ten-fold excess, and reacted for 30min;
(7) Washing: DMF (10 ml/g) once, methanol (10 ml/g) twice, DMF (10 ml/g) twice;
(8) Repeating the steps (2) to (6), and connecting the amino acids in the sequences from right to left;
(9) Cleaving the polypeptide side chain protecting group, and then precipitating the polypeptide with cold diethyl ether;
preparing cutting fluid (10/g): TFA94.5%; 2.5% of water; EDT 2.5%; TIS1%;
cutting time: 120min;
(10) The ether precipitated polypeptide product was dried and then identified, and purified by HPLC (FIG. 4) and Mass identified (FIG. 3) to obtain the polypeptide variant RH-001 of this example.
The synthetic method of the polypeptide modified RH-002 comprises the following steps:
(1) Swelling of the resin: placing 2-Chlorotrityl Chloride Resin resin into a reaction tube, adding DMF (15 ml/g), and oscillating for 30min;
(2) Coupling a first amino acid: filtering out the solvent through a sand core, taking Fmoc-Tyr (tBu) -OH amino acid which is equal to the resin in mole, adding DIEA with 10 times of mole excess and HBTU with three times of mole excess, finally adding DMF for dissolution, and oscillating for 30min;
(3) Deprotection: DMF was removed, 20% piperidine DMF solution (15 ml/g) was added for 5min, and 20% piperidine DMF solution (15 ml/g) was removed for 15min;
(4) And (3) detection: pumping off piperidine solution, taking more than ten resin particles, washing with ethanol for three times, adding ninhydrin, KCN and phenol solution into the resin particles, heating the mixture at 105-110 ℃ for 5min to turn deep blue into positive reaction;
(5) Washing: DMF (10 ml/g) was twice, methanol (10 ml/g) was twice, DMF (10 ml/g) was twice;
(6) Condensation: three times excess of protective amino acid (Fmoc-Leu-OH), three times excess of HBTU, all dissolved with as little DMF as possible, added to the reaction tube, immediately added with ten times excess NMM for 30min;
(7) Washing: DMF (10 ml/g) once, methanol (10 ml/g) twice, DMF (10 ml/g) twice;
(8) Repeating the steps (2) to (6), and connecting the amino acids in the sequences from right to left;
(9) Cleaving the polypeptide side chain protecting group, and then precipitating the polypeptide with cold diethyl ether;
preparing cutting fluid (10/g): TFA94.5%; 2.5% of water; EDT 2.5%; TIS1%;
cutting time: 120min;
(10) The ether precipitated polypeptide product was dried and then identified, and purified by HPLC (FIG. 5) and Mass identified (FIG. 6) to obtain the polypeptide modification RH-002 of this example.
The synthetic method of the polypeptide modified RH-003 comprises the following steps:
(1) Swelling of the resin: placing 2-Chlorotrityl Chloride Resin resin into a reaction tube, adding DMF (15 ml/g), and oscillating for 30min;
(2) Coupling a first amino acid: filtering out the solvent through a sand core, taking Fmoc-Tyr (tBu) -OH amino acid which is equal to the resin in mole, adding DIEA with 10 times of mole excess and HBTU with three times of mole excess, finally adding DMF for dissolution, and oscillating for 30min;
(3) Deprotection: DMF was removed, 20% piperidine DMF solution (15 ml/g) was added for 5min, and 20% piperidine DMF solution (15 ml/g) was removed for 15min;
(4) And (3) detection: pumping off piperidine solution, taking more than ten resin particles, washing with ethanol for three times, adding ninhydrin, KCN and phenol solution into the resin particles, heating the mixture at 105-110 ℃ for 5min to turn deep blue into positive reaction;
(5) Washing: DMF (10 ml/g) was twice, methanol (10 ml/g) was twice, DMF (10 ml/g) was twice;
(6) Condensation: three times excess of protective amino acid (Fmoc-Leu-OH), three times excess of HBTU, all dissolved with as little DMF as possible, added to the reaction tube, immediately added with ten times excess NMM for 30min;
(7) Washing: DMF (10 ml/g) once, methanol (10 ml/g) twice, DMF (10 ml/g) twice;
(8) Repeating the steps (2) to (6), and connecting the amino acids in the sequences from right to left;
(9) Cleaving the polypeptide side chain protecting group, and then precipitating the polypeptide with cold diethyl ether;
preparing cutting fluid (10/g): TFA94.5%; 2.5% of water; EDT 2.5%; TIS1%;
cutting time: 120min;
(10) The ether precipitated polypeptide product was dried and then identified, and subjected to HPLC purification (FIG. 8) and Mass identification (FIG. 7) to obtain the polypeptide modification RH-003 of this example.
The synthetic method of the polypeptide modified RH-004 comprises the following steps:
(1) Swelling of the resin: placing 2-Chlorotrityl Chloride Resin resin into a reaction tube, adding DMF (15 ml/g), and oscillating for 30min;
(2) Coupling a first amino acid: filtering out the solvent by sand core suction, taking Fmoc-Val-OH amino acid with the same mole as resin, adding 10 times of excessive DIEA, three times of HBTU, adding DMF to dissolve, and oscillating for 30min;
(3) Deprotection: DMF was removed, 20% piperidine DMF solution (15 ml/g) was added for 5min, and 20% piperidine DMF solution (15 ml/g) was removed for 15min;
(4) And (3) detection: pumping off piperidine solution, taking more than ten resin particles, washing with ethanol for three times, adding ninhydrin, KCN and phenol solution into the resin particles, heating the mixture at 105-110 ℃ for 5min to turn deep blue into positive reaction;
(5) Washing: DMF (10 ml/g) was twice, methanol (10 ml/g) was twice, DMF (10 ml/g) was twice;
(6) Condensation: triple excess of protected amino acid (Fmoc-Lys (Boc) -OH), triple excess of HBTU, all dissolved with as little DMF as possible, added to the reaction tube, immediately added to NMM ten-fold excess, and reacted for 30min;
(7) Washing: DMF (10 ml/g) once, methanol (10 ml/g) twice, DMF (10 ml/g) twice;
(8) Repeating the steps (2) to (6), and connecting the amino acids in the sequences from right to left;
(9) Cleaving the polypeptide side chain protecting group, and then precipitating the polypeptide with cold diethyl ether;
preparing cutting fluid (10/g): TFA94.5%; 2.5% of water; EDT 2.5%; TIS1%;
cutting time: 120min;
(10) The ether precipitated polypeptide product was dried and then identified, and subjected to HPLC purification (FIG. 9) and Mass identification (FIG. 10) to obtain the polypeptide modification RH-004 of this example.
The synthetic method of the polypeptide modified bodies RH-M-001, RH-M-002, RH-M-003 and RH-M-004 comprises the following steps:
(1) Assembling a synthetic peptide chain according to the above-mentioned RH-001 synthesis step;
(2) After the peptide chain is assembled, adding excessive myristic acid to condensation couple the myristic acid to the N end of the polypeptide chain;
(3) Cleaving the polypeptide side chain protecting group, and then precipitating the polypeptide with cold diethyl ether;
preparing cutting fluid (10/g): TFA94.5%; 2.5% of water; EDT 2.5%; TIS1%;
cutting time: 120min;
(4) The ether precipitated polypeptide product was blow dried and then identified, purified by HPLC (FIG. 12) and Mass identified (FIG. 11) to obtain polypeptide modification RH-M-001.
Polypeptide modifications RH-M-002 (HPLC purification results are shown in FIG. 13, mass identification results are shown in FIG. 14), RH-M-003 (HPLC purification results are shown in FIG. 16, mass identification results are shown in FIG. 15) and RH-M-004 (HPLC purification results are shown in FIG. 17, mass identification results are shown in FIG. 18) were obtained by the same method as described above.
Patentees continue to test the biosafety and bioavailability of the polypeptide variants obtained above.
First, each polypeptide variant was analyzed for biosafety by CCK8 cytotoxicity assay. The testing method comprises the following steps:
bone marrow-derived macrophages (BMDM) suspensions were seeded in 96-well plates at 100. Mu.l per well, 0. Mu.M (control group), 3.125. Mu.M, 6.25. Mu.M, 12.5. Mu.M, 25. Mu.M, 50. Mu.M polypeptide RH-000 and each polypeptide variant were added to each well cell suspension according to a concentration gradient, and after incubation for 24 hours, cell viability was measured using an enzyme-labeled instrument (experiment was repeated 3 times). The test results are shown in fig. 19 to 26.
As can be seen from fig. 19 to fig. 26, the cell activity of each polypeptide modification treatment group was not significantly reduced with the increase of the drug concentration compared with the control group, indicating that each polypeptide modification prepared by the present invention has no cytotoxicity and high biosafety.
Then testing the sepsis treatment effect of the polypeptide RH-000 and each polypeptide modification to analyze the bioavailability, wherein the testing method comprises the following steps:
taking polypeptide RH-000 and polypeptide modified bodies, sequentially arranging corresponding test groups of 9 mice each, and respectively performing intraperitoneal injection of 8×10 6 Coli of CFU, then given polypeptide RH-000 and single dose therapy of each polypeptide variant (10 mg/kg per mouse), respectively; setting a control group at the same time, and giving the same dosage of physiological saline to the control group; the therapeutic effect of polypeptide RH-000 and each polypeptide variant in sepsis mice was analyzed by Kaplan-Meier survival curves, and the analysis results are shown in FIGS. 27 and 28.
As shown in FIG. 27, the polypeptide RH-000 and the modified polypeptide RH-001, RH-002, RH-003 and RH-004 can significantly improve the survival rate of mice compared with the control group. Wherein, the survival rate of 72h of the RH-000 treatment group sepsis mice is 56%, and the survival rates of 72h of the RH-001 and RH-002 treatment group sepsis mice respectively reach 89% and 67%, which is obviously better than that of the RH-000 treatment group; the survival rate of the sepsis mice in the polypeptide modified RH-003 treatment group is always kept at 100%, which is obviously superior to that of the control group and other treatment groups.
As shown in FIG. 28, the polypeptide RH-000 and the modified polypeptide RH-M-001, RH-M-002, RH-M-003 and RH-M-004 can significantly improve the survival rate of mice compared with the control group. Wherein the 72h survival rate of the polypeptide modified RH-M-001 and RH-M-002 and the polypeptide RH-000 treatment group to mice is between 50 and 70 percent; the survival rate of mice in the polypeptide modified RH-M-004 treatment group is always kept at 100%, which is obviously superior to that of the control group and other treatment groups.
Notably, 12h survival rates of RH-M-003 treated groups are better than RH-003 treated groups after modification of myristic acid at the N-terminus of the polypeptide, 36h survival rates of RH-M-001 and RH-M-002 treated groups are better than RH-001 and RH-002 treated groups, respectively, and 72h survival rates of RH-M-004 treated groups are much better than RH-004 treated groups, indicating that modification of myristic acid at the N-terminus of the polypeptide is beneficial for improving survival rates of mice. This is probably because the penetration and stability of the modified polypeptide variant are improved, so that the polypeptide variant can exert its intended effect.
On the basis of biological safety and biological effectiveness, the polypeptide with fewer amino acid numbers and less modification is selected, so that the success rate and purity of the synthesis of the polypeptide can be improved, the production cost can be reduced, and the preparation economy can be improved. On the premise, RH-001, RH-002, RH-003 and RH-M-004 are selected as post-research medicines, and the influence of medicine treatment on the bacterial load in the mice is tested.
The testing method comprises the following steps: five test groups and a control group of 6 mice were set, each by intraperitoneal injection of 8X 10 6 Coli of CFU was then given single dose therapy (10 mg/kg per mouse) with physiological saline (control group), RH-001, RH-002, RH-003 and RH-M-004, respectively. After 8 hours of treatment, respectively collecting samples of blood, liver, spleen, lung and kidney, and quantifying bacteria; the results are shown in fig. 29, 30, 31, 32 and 33.
As can be seen from FIGS. 29, 30, 31, 32 and 33, RH-001, RH-002, RH-003 and RH-M-004 are each effective in reducing bacterial load of blood and organs.
Claims (4)
1. A polypeptide variant comprising the amino acid sequence set forth in seq id no:
Gly 1 -Val 2 -Lys 3 -Leu 4 -Tyr 5 -Gly 6 -Ser 7 -Asp 8 -Lys 9 -Ser 10 。
2. the polypeptide modification of claim 1, wherein the free carboxyl group of the N-terminal amino acid is linked to a fatty acid by a dehydration condensation reaction.
3. The polypeptide variant of claim 2, wherein the fatty acid is myristic acid, lauric acid or palmitic acid.
4. Use of a polypeptide modification according to any one of claims 1-3 for the preparation of an anti-inflammatory and bactericidal medicament, wherein the anti-inflammatory and bactericidal medicament is an anti-escherichia coli infection medicament.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202310942038.XA CN117024564A (en) | 2021-11-17 | 2021-11-17 | Polypeptide modified body and application thereof in preparation of anti-inflammatory and bactericidal medicines |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202111359409.9A CN114195858B (en) | 2021-11-17 | 2021-11-17 | Polypeptide and modified body thereof and application of modified body in preparation of anti-inflammatory and bactericidal medicines |
CN202310942038.XA CN117024564A (en) | 2021-11-17 | 2021-11-17 | Polypeptide modified body and application thereof in preparation of anti-inflammatory and bactericidal medicines |
Related Parent Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202111359409.9A Division CN114195858B (en) | 2021-11-17 | 2021-11-17 | Polypeptide and modified body thereof and application of modified body in preparation of anti-inflammatory and bactericidal medicines |
Publications (1)
Publication Number | Publication Date |
---|---|
CN117024564A true CN117024564A (en) | 2023-11-10 |
Family
ID=80647801
Family Applications (4)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202310942038.XA Pending CN117024564A (en) | 2021-11-17 | 2021-11-17 | Polypeptide modified body and application thereof in preparation of anti-inflammatory and bactericidal medicines |
CN202310942039.4A Pending CN117024565A (en) | 2021-11-17 | 2021-11-17 | Polypeptide modified body and application thereof in preparation of anti-escherichia coli infection drugs |
CN202311068041.XA Pending CN118255838A (en) | 2021-11-17 | 2021-11-17 | Polypeptide modified body and application thereof in preparation of anti-inflammatory and bactericidal medicines |
CN202111359409.9A Active CN114195858B (en) | 2021-11-17 | 2021-11-17 | Polypeptide and modified body thereof and application of modified body in preparation of anti-inflammatory and bactericidal medicines |
Family Applications After (3)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202310942039.4A Pending CN117024565A (en) | 2021-11-17 | 2021-11-17 | Polypeptide modified body and application thereof in preparation of anti-escherichia coli infection drugs |
CN202311068041.XA Pending CN118255838A (en) | 2021-11-17 | 2021-11-17 | Polypeptide modified body and application thereof in preparation of anti-inflammatory and bactericidal medicines |
CN202111359409.9A Active CN114195858B (en) | 2021-11-17 | 2021-11-17 | Polypeptide and modified body thereof and application of modified body in preparation of anti-inflammatory and bactericidal medicines |
Country Status (1)
Country | Link |
---|---|
CN (4) | CN117024564A (en) |
Family Cites Families (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP1481680A1 (en) * | 2003-05-30 | 2004-12-01 | Aventis Pharma Deutschland GmbH | Use of S1P |
EP1814911A2 (en) * | 2004-11-04 | 2007-08-08 | New England Medical Center Hospitals, Inc. | G protein coupled receptor agonists and antagonists and methods of use |
WO2015109112A1 (en) * | 2014-01-16 | 2015-07-23 | Gemmus Pharma Inc. | Combination treatment of antibiotic and gpcr agonist for viral/bacterial co-infections |
CN106344922A (en) * | 2016-09-14 | 2017-01-25 | 安徽济人药业有限公司 | Application of G protein coupling receptor 18 agonist in preparation of infection drugs |
CN107469067B (en) * | 2017-09-05 | 2018-06-19 | 浙江大学 | A kind of application of polypeptide and its variant in immunoregulation medicament is prepared |
US20210382068A1 (en) * | 2018-10-02 | 2021-12-09 | Dana-Farber Cancer Institute, Inc. | Hla single allele lines |
CN114181293B (en) * | 2021-12-06 | 2023-09-22 | 郑州大学 | Humanized antibacterial peptide LL-37 modified body and application thereof |
-
2021
- 2021-11-17 CN CN202310942038.XA patent/CN117024564A/en active Pending
- 2021-11-17 CN CN202310942039.4A patent/CN117024565A/en active Pending
- 2021-11-17 CN CN202311068041.XA patent/CN118255838A/en active Pending
- 2021-11-17 CN CN202111359409.9A patent/CN114195858B/en active Active
Also Published As
Publication number | Publication date |
---|---|
CN117024565A (en) | 2023-11-10 |
CN114195858A (en) | 2022-03-18 |
CN118255838A (en) | 2024-06-28 |
CN114195858B (en) | 2023-09-08 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN104662038B (en) | Glucagon analogue | |
CN106632682A (en) | Fusion protein IFN-ELP and application thereof | |
CN1321688C (en) | Hemoglobin alpha chain peptide fragments useful for inhibiting stem cell proliferation | |
CN104530199B (en) | A kind of tumor protein p53 and its preparation method and application | |
CN102827284A (en) | Exendin or analogue containing polyethylene glycol group and preparation and application thereof | |
TWI588153B (en) | Polypeptides, nucleic acid molecule encoding polypeptides, and uses of polypeptides | |
CN115260289B (en) | Inflammation colon targeting peptide and screening method thereof | |
ES2272325T3 (en) | PEPTIDES AGAINST CAUSING AUTHORTIC BODIES OF CMD. | |
KR20190067219A (en) | Use of neurotoxic injury-related polypeptides in pain prevention, relief or treatment | |
CN109718363A (en) | Prevention, the peptide for alleviating or treating Alzheimer disease and its application | |
CN110179994A (en) | A kind of temperature and enzyme dual responsiveness protein high molecular conjugate and the preparation method and application thereof | |
CN101041693B (en) | Novel blood sugar lowing polypeptide and uses thereof | |
EP2799445A1 (en) | Integrin blocker polypeptide and use thereof | |
CN107760661A (en) | PEG trims of medicinal kininogenase and its preparation method and application | |
EP1597271A2 (en) | Peptides directed against antibodies, which cause cold-intolerance, and the use thereof | |
ES2197840T3 (en) | PROCEDURE FOR THE OBTAINING AND USE OF NEW HUMAN DEFENSINS AS BIOLOGICALLY ACTIVE PROTEINS FOR THE TREATMENT OF INFECTIONS AND OTHER DISEASES. | |
CN114195858B (en) | Polypeptide and modified body thereof and application of modified body in preparation of anti-inflammatory and bactericidal medicines | |
CN107805282A (en) | A kind of targeted therapies and the united polypeptide of immunotherapy | |
CN109069581A (en) | Lock ring helical peptides and synthetic method | |
CN101280012B (en) | Exendin-4 active isomer and application thereof | |
CN107216369A (en) | Similar active peptide of a kind of nucleoprotamine and preparation method and application | |
CN100334114C (en) | Novel fusion protein production and uses | |
CN103012555A (en) | Preparation method for new polypeptide having tissue protection activity, and application of new polypeptide in treatment | |
ES2239814T3 (en) | METHOD OF MOBILIZATION OF HEMATOPOYETIC MOTHER CELLS. | |
EP4386008A1 (en) | New mutant of recombinant ganoderma lucidum immunoregulatory protein and application thereof |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination |