Vaccine for preventing novel coronavirus pneumonia COVID-19 and preparation method thereof

A technology of CRM197, SEQIDNO.1-SEQIDNO.11, which is applied in the field of vaccines and preparations for the prevention of novel coronavirus pneumonia COVID-19, can solve the problems such as the loss of efficacy of biohazardous original vaccines, and reduce research and development costs and research and development cycles , enhance the immune response of the human body, and facilitate large-scale mass production

Pending Publication Date: 2022-02-18
LIAONING CHENGDA BIOTECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Traditional vaccines are made by inactivating or attenuating pathogens, and there are problems such as biological hazards and genetic variation that cause the original vaccine to lose efficacy

Method used

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  • Vaccine for preventing novel coronavirus pneumonia COVID-19 and preparation method thereof
  • Vaccine for preventing novel coronavirus pneumonia COVID-19 and preparation method thereof
  • Vaccine for preventing novel coronavirus pneumonia COVID-19 and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Example 1 Synthetic method of target polypeptide.

[0061] 1. The polypeptide whose amino acid sequence is shown in SEQ ID NO.1-SEQNO.11 was prepared by conventional solid-phase polypeptide synthesis method, and the specific steps are as follows.

[0062] (1) Remove Fmoc protection.

[0063] Fill a commercially available Rink Amide AM resin into an organic-solvent-resistant reaction tube with a filter and cap tightly with an organic-solvent-resistant cap. After washing with DMF (dimethylformamide) for 1 minute, add an excess of 20% piperidine / DMF (volume ratio) solution, cap tightly and shake the reaction tube gently to mix it evenly and keep the Fmoc protection removed for 15 minutes . After draining, wash three times with DMF.

[0064] (2) Peptide bond condensation.

[0065] The Fmoc-protected amino acid of 3 times the molar weight of the resin amino group, the activator HBTU of 3 times the molar equivalent of the resin amino group, and the DIPEA (N , N-diisoprop...

Embodiment 2

[0077]Example 2 Preparation method of immunogenic conjugate.

[0078] The list of conjugates formed by the synthetic peptides (S1-S11) obtained in Example 1 with CRM197, TT, DT, and M-M-OMP respectively through SMCC LINKER or DSAP LINKER is shown in Table 2.

[0079] TT tetanus toxin protein and tetanus toxin can be obtained from Bacillus tetani through fermentation, lysis, centrifugation and chromatographic purification.

[0080] DT diphtheria toxin protein and diphtheria toxin carrier protein DT can be obtained from diphtheria bacillus through fermentation, cracking, centrifugation and chromatography purification.

[0081] CRM197 Recombinant Diphtheria Toxin Protein, CRM197 is a diphtheria bacillus reconstructed from the gene sequence, which is obtained through fermentation, lysis, centrifugation, and chromatography purification.

[0082] Meningococcal outer membrane protein (M-OMP), obtained from meningococcus, is fermented, lysed, centrifuged, and chromatographically puri...

Embodiment 3 4

[0098] Example 3 Preparation and immune effect of tetravalent peptide vaccine.

[0099] 1. Vaccine preparation.

[0100] 1. Materials.

[0101] The polypeptide conjugate prepared in Example 2.

[0102] Adjuvants: aluminum hydroxide (Al(OH)3), aluminum phosphate (AlPO4), MPL, CpG.

[0103] 2. Preparation method.

[0104] Select the immunogenic conjugates combined with the synthetic peptides in Example 2 and different carrier proteins, preferably different immunogenic conjugates with sequences SEQID NO.4, 5, 6, and 7, and combine them with different adjuvants to prepare four For peptide vaccines, the total concentration of the four peptides is 100ug / ml, and the specific combination scheme is shown in Table 3.

[0105] Table 3. Cross-combination scheme of polypeptide conjugates and adjuvants.

[0106]

[0107] Note: Combination serial number naming rules: arrange the conjugates according to the size of the number, "conjugate carrier initial letter" + "adjuvant number".

...

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Abstract

The invention relates to the fields of immunological technology, biotechnology and biological medicine, in particular to a vaccine for preventing novel coronavirus pneumonia COVID-19 and a preparation method thereof. A composition of immunogenic conjugates includes at least two combinations of immunogenic conjugates containing the same carrier protein or at least two combinations of immunogenic conjugates containing two different carrier proteins. The composition of the immunogenic conjugates is used for preparing a vaccine for preventing COVID-19. Compared with a traditional attenuated vaccine, an inactivated vaccine and other novel vaccines, the vaccine can quickly cope with virus variation; the field requirement for vaccine production is reduced, and large-scale batch production is easy to realize; the research and development cost and the research and development period are reduced; safety, no toxicity and stability are realized; as the molecular structure is small and simple, the problems of serious complications and iatrogenic infection are rarely caused.

Description

technical field [0001] The invention relates to the fields of immunology technology, biotechnology and biomedicine, in particular to a vaccine for preventing novel coronavirus pneumonia COVID-19 and a preparation method thereof. Background technique [0002] Coronavirus is a positive enveloped virus with RNA, and its genome size is about 26-32kb, which is the largest RNA virus known genome. Genomic RNA and phosphorylated nucleocapsid (N) proteins are buried in the phospholipid bilayer and covered by the spike glycoprotein trimer (S), the membrane (M) protein (type III transmembrane glycoprotein) and the envelope ( E) The protein is located between the S proteins of the viral envelope. Coronaviruses have a variety of hosts, including birds and mammals, especially bats. Coronavirus is a kind of virus that widely exists in nature and can cause multi-system diseases including respiratory tract, digestive tract and nervous system. Highly pathogenic coronavirus infection has bec...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/215A61P31/14A61P11/00A61K39/39A61K39/385
CPCA61K39/12A61K39/39A61P31/14A61P11/00A61K39/385A61K2039/6037A61K2039/6068A61K2039/575
Inventor 张庶民刘刚高军毛昱于文俊周荔葆孙韦强孙述学车兴华
Owner LIAONING CHENGDA BIOTECH
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