CN115845039A - Broad-spectrum polypeptide tumor vaccine composition - Google Patents
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Abstract
The invention discloses a broad-spectrum polypeptide tumor vaccine composition, discloses one or more nucleic acid molecules of a nucleic acid sequence for coding a mutated WT1 antigen, comprises a vector, a composition and a vaccine of the nucleic acid sequence of a WT1 target antigen, a MUC1 target antigen and a PD-1 target antigen, and discloses the mutated WT1 target antigen, the MUC1 target antigen and the PD-1 target antigen.
Description
Technical Field
The invention relates to the field related to broad-spectrum polypeptide tumor vaccines, in particular to a composition of a broad-spectrum polypeptide tumor vaccine.
Background
The cancer vaccine market is growing rapidly, with lymphoma vaccines accounting for about 0.5% of the market share, effective tumor vaccines can be used for more effective, less toxic alternative treatments to prevent tumor growth and/or can be used as standard treatments for patients with advanced cancer, the cancer-associated antigen and thus the target for anti-tumor vaccines is WT1;
wilms tumor suppressor gene 1 (WT 1) was identified as the cause of renal embryonic malignancy, affecting about 1/10,000 infants, which occurred in sporadic and hereditary forms, inactivation of WT1 resulted in the development of wilms tumors and Denys-Drash syndrome (DDS), with consequent renal disease and possible genital abnormalities, WT1 protein was found to interact with a number of cytokines, including the major tumor regulatory gene p53, which p53 is also a tumor suppressor transcription factor;
the WT1 gene is expressed in many tumor types and has been more widely involved in many cancers, for example, WT1 protein has been localized in the nucleus of 75% of mesothelioma (14,200 cases and 93% of ovarian serous carcinoma worldwide (190,000 cases of ovarian cancer worldwide in 2010)) and, in addition, WT1 has been involved in pancreatic cancer, leukemia, lung cancer, breast cancer, colon cancer, glioblastoma, head and neck cancer, and benign mesothelioma and cervical and ovarian cancers, etc., WT1 being a target of gene therapy or immunotherapy as a method of cancer treatment;
WT1 encodes a transcription factor containing 4 zinc finger motifs on the C-terminus and a proline/glutamine rich DNA binding domain on the N-terminus, which plays an essential role in the normal development of the urogenital system, however, it is less essential in adults, suggesting that it is a target for immunotherapy, multiple transcript variants resulting from alternative splicing on two coding exons are well characterized and maximizing CTL coverage using the entire reading frame would be considered advantageous;
most of the efforts to generate strong immunity against this gene target have not been successful due to the conservation of WT1 antigen, and vaccines have been previously studied using DAN vaccine technology, poxvirus vaccine technology, adenovirus vaccine technology, peptide vaccine technology, and protein-based vaccine technology, using the true gene structure, i.e., the native "normal" genes, in which only low-level or non-functional T cell immunity is achieved;
vaccines for the treatment and prevention of cancer are of great interest, and existing WT 1-targeting vaccines are limited by poor antigen expression in vivo, and therefore there remains a need in the art to develop safe and effective vaccines suitable for WT 1-expressing tumors, thereby providing treatment of such cancers and improving the survival rate of said cancers.
Disclosure of Invention
Accordingly, in order to address the above deficiencies, the present invention herein provides a broad spectrum polypeptide tumor vaccine composition, preferably, a broad spectrum polypeptide antigen such as one which encodes at least one amino acid sequence selected from the group consisting of SEQ ID NO. 1 and SEQ ID NO. 3.
Preferably, the amino acid sequences comprise the respective combinations of the sequences SEQ ID NO. 1, 2 and 3 or two of the respective combinations of the sequences.
Preferably, the broad spectrum polypeptide antigen consists of a WT1 target antigen, a MUC1 target antigen and a PD-1 target antigen.
Preferably, the constituent nucleic acid molecules incorporate the molecules into a plasmid or viral vector.
Preferably, the use of the constitutively isolated nucleic acid molecule in the manufacture of a medicament for inducing an immune response against WT1 in an individual.
Preferably, the composition comprises a fusion protein, the recombinant plasmid is transformed into a cell, and the fusion protein is obtained by culturing and IPTG induction, wherein the fusion protein comprises a series arrangement of sequences of broad-spectrum polypeptide antigens.
Preferably, the broad-spectrum polypeptide tandem fusion protein and the broad-spectrum polypeptide tandem fusion obtained by the construction method are fused in series.
Preferably, the isolated nucleic acid molecule comprising one or more nucleic acid sequences selected from the group consisting of a nucleic acid sequence encoding seq id No. 1, a nucleic acid sequence encoding a fragment comprising at least 90% of the length of seq id No. 1, a nucleic acid sequence encoding a protein having at least 98% identity to seq id No. 1, a nucleic acid sequence encoding a fragment comprising at least 90% of the length of a protein having at least 98% identity to seq id No. 1, a nucleic acid sequence encoding seq id No. 3, a nucleic acid sequence encoding a fragment comprising at least 90% of the length of seq id No. 3, a nucleic acid sequence encoding a protein having at least 97% identity to seq id No. 3, and a nucleic acid sequence encoding a fragment comprising at least 90% of the length of a protein having at least 97% identity to seq id No. 3.
Preferably, the present invention also relates to an isolated nucleic acid molecule comprising one or more nucleic acid sequences selected from the group consisting of: SEQ ID NO 2, a fragment comprising at least 85% of the length of SEQ ID NO 1, a nucleic acid sequence having at least 85% identity to SEQ ID NO 2, a fragment comprising at least 85% of the length of a nucleic acid sequence having at least 98% identity to SEQ ID NO 2, SEQ ID NO 3, a fragment comprising at least 85% of the length of SEQ ID NO 3, a nucleic acid sequence having at least 95% identity to SEQ ID NO 3, and a fragment comprising at least 80% of the length of a nucleic acid sequence having at least 98% identity to SEQ ID NO 3.
Preferably, the invention also relates to a vaccine comprising a nucleic acid molecule which may comprise a nucleic acid sequence having at least about 90% identity over the entire length of the nucleic acid sequence shown in seq id No. 1, which may comprise a nucleic acid sequence having at least about 85% identity over the entire length of the nucleic acid sequence shown in seq id No. 3, and a peptide which may comprise an amino acid sequence having at least about 90% identity over the entire length of the amino acid sequence shown in seq id No. 2, which may comprise an amino acid sequence having at least about 90% identity over the entire length of the amino acid sequence shown in seq id No. 3.
Preferably, the nucleic acid molecules described above may be incorporated into a plasmid or viral vector, the invention also relates to compositions comprising one or more of the nucleic acid molecules described above, and the invention also relates to vaccines comprising one or more of the nucleic acid molecules described above.
Preferably, the nucleic acid molecule may comprise an expression vector, the vaccine may further comprise a pharmaceutically acceptable excipient, and the vaccine may further comprise an adjuvant.
Preferably, the present invention also relates to a peptide comprising the amino acid sequence shown in SEQ ID NO 1
Preferably, the present invention also relates to a peptide comprising the amino acid sequence shown in SEQ ID NO 2.
Preferably, the present invention also relates to a peptide comprising the amino acid sequence shown in SEQ ID NO 3.
Drawings
FIG. 1 is a graph showing a plot of an immune group versus plaque forming units (SFU) per 106 splenocytes of the invention;
FIG. 2 is a graph showing the SFU of a plotted immune group versus each 106 splenocytes according to the present invention;
FIG. 3 shows a nucleotide sequence encoding WT1 according to the present invention;
FIG. 4 is a nucleotide sequence showing the encoding of Muc1 according to the invention;
FIG. 5 is a nucleotide sequence of the present invention showing the encoding of PD-1.
Detailed Description
The present invention will be described in detail with reference to fig. 1 to 5, and the technical solutions in the embodiments of the present invention will be clearly and completely described, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
1. Definition of
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art and, in case of conflict, the present document (including definitions) controls that the preferred methods and materials are described below, although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, all publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety and the materials, methods, and examples disclosed herein are illustrative only and not intended to be limiting.
As used herein, the terms "comprises/comprising," "includes," "having," "has," "can," "containing," and variations thereof, are intended to be open-ended transition phrases, terms, or words that do not exclude the possibility of additional acts or structures, the singular forms "a," "an," and "the" include plural references unless the context clearly dictates otherwise, and the disclosure also encompasses "comprising/including" the embodiments or components set forth herein, "consisting of" the embodiments or components, and "other embodiments consisting essentially of" the embodiments or components, whether or not explicitly shown.
As used herein, "adjuvant" can mean any molecule added to the DNA plasmid vaccines described herein to enhance the antigenicity of one or more antigens encoded by the DNA plasmids and encoding nucleic acid sequences described below.
The antigen "means: a protein having a WT1 amino acid sequence comprising a mutation of seq id No. 1; fragments thereof having the length shown herein, variants, i.e. proteins having a sequence with identity to seq id No. 1 as shown herein, fragments of variants having the length shown herein, seq id No. 3; fragments thereof having the lengths shown herein, variants, i.e., proteins having a sequence with the identity of seq id No. 3 as shown herein, fragments of variants having the lengths shown herein, and combinations thereof, the antigen may optionally comprise signal peptides such as those from other proteins.
The vaccine can be administered to a mammal to elicit an immune response in the mammal. The mammal can be a human, primate, non-human primate, cow, sheep, goat, antelope, bison, buffalo, bison, bovidae, deer, hedgehog, elephant, camel, alpaca, mouse, rat, chicken, and pig.
"fragment" may mean a polypeptide fragment of an antigen capable of eliciting an immune response in a mammal, which fragment may be 100% identical to full length in each case with or without a signal peptide and/or methionine at position 1, which fragment may comprise 20% or more, 25% or more, 30% or more, 35% or more, 40% or more, 45% or more, 50% or more, 55% or more, 60% or more, 65% or more, 70% or more, 75% or more, 80% or more, 85% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more of the length of a particular full-length antigen, excluding any added heterologous signal peptide, fragment of a polypeptide, which fragment has 95% or more, 96% or more, 97% or more, 98% or more, 99% or more of the same signal peptide as the antigen, and/or a signal peptide at position 1, and/or a methionine at the N-terminal may comprise an additional percentage of the same signal peptide, such as the signal peptide, and/or a signal peptide, which may comprise additional amino acid, such as an immunoglobulin, and/or a signal peptide, which may comprise an additional amino acid, which is not be added to the same, and/or a signal peptide.
In each case with or without a sequence encoding a signal peptide and/or a methionine at position 1, a fragment of the nucleic acid sequence encoding the antigen may be 100% identical to full length, may comprise 20% or more, 25% or more, 30% or more, 35% or more, 40% or more, 45% or more, 50% or more, 55% or more, 60% or more, 65% or more, 70% or more, 75% or more, 80% or more, 85% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more of the length of the particular full-length coding sequence, excluding any added heterologous signal peptide, a fragment encoding a polypeptide having a methionine of 95% or more, 96% or more, 97% or more, 98% or more, 99% or more, or a sequence encoding a signal peptide, and/or optionally may comprise a percentage of the same amino acid sequence as the full-length encoding peptide, except when the sequence is missing at the end of the sequence encoding the signal peptide and/or methionine.
The basic principles and main features of the present invention and the advantages of the present invention have been shown and described, and the standard parts used in the present invention are all available on the market, the special-shaped parts can be customized according to the description and the accompanying drawings, the specific connection mode of each part adopts the conventional means of bolt and rivet, welding and the like mature in the prior art, the machinery, parts and equipment adopt the conventional type in the prior art, and the circuit connection adopts the conventional connection mode in the prior art, and the details are not described herein.
The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
Claims (7)
1. A broad-spectrum polypeptide tumor vaccine composition, which is characterized in that: the broad spectrum polypeptide antigen encodes at least one amino acid sequence selected from the group consisting of SEQ ID NO 1 and SEQ ID NO 3.
2. The broad-spectrum polypeptide tumor vaccine composition of claim 1, wherein the amino acid sequence comprises the sequences of SEQ ID NOs 1, 2 and 3 or any one of the two sequences.
3. The broad-spectrum polypeptide tumor vaccine composition of claim 1, wherein the broad-spectrum polypeptide antigen consists of a WT1 target antigen, a MUC1 target antigen and a PD-1 target antigen.
4. A broad spectrum polypeptide tumor vaccine composition according to claim 1, wherein said nucleic acid molecule is incorporated into a plasmid or viral vector.
5. A broad spectrum polypeptide tumor vaccine composition of claim 1, the use of the isolated nucleic acid molecule of the composition in the preparation of a medicament for inducing an immune response against WT1 in an individual.
6. The broad-spectrum polypeptide tumor vaccine composition of claim 1, which comprises fusion protein, wherein the fusion protein is obtained by transforming cells with recombinant plasmid, culturing and inducing with IPTG, and the fusion protein comprises a series arrangement of broad-spectrum polypeptide antigens.
7. The broad-spectrum polypeptide tumor vaccine composition according to claim 1 or 5, wherein the broad-spectrum polypeptide tandem fusion protein is fused with the broad-spectrum polypeptide tandem fusion protein constructed by the construction method.
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| CN202210766801.3A CN115845039A (en) | 2022-07-01 | 2022-07-01 | Broad-spectrum polypeptide tumor vaccine composition |
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| CN202210766801.3A CN115845039A (en) | 2022-07-01 | 2022-07-01 | Broad-spectrum polypeptide tumor vaccine composition |
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104812401A (en) * | 2012-12-13 | 2015-07-29 | 宾夕法尼亚大学理事会 | Wt1 vaccine |
| CN105025932A (en) * | 2013-03-15 | 2015-11-04 | 宾夕法尼亚大学理事会 | Cancer vaccines and methods of treatment using the same |
| CN113603784A (en) * | 2015-05-29 | 2021-11-05 | 艾吉纳斯公司 | anti-CTLA-4 antibodies and methods of use thereof |
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- 2022-07-01 CN CN202210766801.3A patent/CN115845039A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104812401A (en) * | 2012-12-13 | 2015-07-29 | 宾夕法尼亚大学理事会 | Wt1 vaccine |
| CN105025932A (en) * | 2013-03-15 | 2015-11-04 | 宾夕法尼亚大学理事会 | Cancer vaccines and methods of treatment using the same |
| CN113603784A (en) * | 2015-05-29 | 2021-11-05 | 艾吉纳斯公司 | anti-CTLA-4 antibodies and methods of use thereof |
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