WO2005023853A1 - プロテアーゼ耐性seb改変体およびそれを含むワクチン - Google Patents
プロテアーゼ耐性seb改変体およびそれを含むワクチン Download PDFInfo
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- WO2005023853A1 WO2005023853A1 PCT/JP2004/012545 JP2004012545W WO2005023853A1 WO 2005023853 A1 WO2005023853 A1 WO 2005023853A1 JP 2004012545 W JP2004012545 W JP 2004012545W WO 2005023853 A1 WO2005023853 A1 WO 2005023853A1
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- seb
- variant
- modified
- amino acid
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Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
- C07K14/305—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Micrococcaceae (F)
- C07K14/31—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Micrococcaceae (F) from Staphylococcus (G)
-
- 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
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/08—Antiallergic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
Definitions
- the present invention provides a Staphylococcus aureus intestinal endotoxin B (known as one of superantigens)
- Staphylococcal enterotoxin B (hereinafter sometimes referred to as “SEB”) or a derivative thereof, a vaccine containing the same as an active ingredient, and use thereof. More specifically, the present invention relates to a modified SEB or a derivative thereof having resistance to proteases and, as a result, reduced toxicity, a peptide containing the same as an active ingredient and a use thereof.
- SEB is a type of enterotoxin (enteric toxin; causative toxin-type food poisoning toxin) produced by Staphylococcus aureus.
- SEB consists of 239 amino acid residues, and its amino acid sequence is also known (SEQ ID NO: 1).
- the SEB molecule is composed of two domains, the first domain consisting of residues 1-120, and the second domain consisting of residues 127-239.
- Staphylococcus aureus is an indigenous bacterium, it is known that infections caused by Staphylococcus aureus, which is resistant to many antibiotics, are extremely serious and have a poor prognosis. Severe staphylococcal infections, as typified by food poisoning, are mainly caused by toxins released from the cells. Among them, Staphylococcal enterotoxin (Staphylococcal enterotoxin, sometimes abbreviated as “SE”) is a type of superantigen that acts on many T lymphocytes and produces a large amount of inflammation that does not normally occur Promotes the production of site force-in (Non-Patent Document 1).
- SE Staphylococcal enterotoxin
- Non-patent Document 2 Non-patent Document 2
- SEB Staphylococcus aureus intestinal endotoxin B
- Non-patent Document 3 In addition, it has been reported that a high proportion of atopic patients have an IgE-type anti-SEB antibody, and a relationship between SEB and pathogenesis is suspected (Non-patent Document 3). [0005] Furthermore, in rheumatic patients, epidemiological data suggesting a relationship between SEB and onset and pathogenesis has been reported, and the relationship between IgM-type anti-SEB antibodies and pathology has been reported (Non-patent Document 4). ).
- Non-Patent Document 7 So far, from the analysis of the three-dimensional structure of SEB and the like, a variant in which the contact site with MHC has been modified, a variant in which the contact site with TCR has been modified, and the like have been reported (Non-Patent Document 7).
- Patent document 1 WO99 / 40935
- Non-Patent Document 1 V. Micusan and J. Thibodeau, "Seminars in Immunol 1 ", 1993, Vol. 5, p. 3-11
- Non-Patent Document 2 Kuwahata, ⁇ ., Et al., "Acta Pediatnca Japonica", 199b, 38, ⁇ .1-Non-Patent Document 3: Zone (Sohn MH.), Kim ( Kim GH), Kim (Kim WK), Young (Jang GC), Kim (Kim KE), "Allergy Asthma Pro", 2003, 24 (1), p.67-71
- Non-Patent Document 4 Origuchi T., Eguchi K., Kawabe Y., Yamashita I., Mizokami A., Ida H. , Nagataki S., "Annual Rheum. Dis.”, 1995, 54 (9), p.713-720
- Non-Patent Document 5 Kuwahata, M., Imanaka, H., Takei, S., and Masuda, K., "Acta Oediatrica Japonica” , 1996, 38, ⁇ .1-7
- Non-Patent Document 6 Puddy (Woody MA), Krakauer T, Styles (Stiles BG), "Vaccine”, 1997, 15 (2), ⁇ ⁇ 133-139
- Non-Patent Document 7 Leader (Leder L,) et al., "Journal of Experimental Medicine", 1998, 187 (6), p.823-833.
- the present inventors have conducted extensive studies on the change in activity caused by amino acid mutations in SEB, and as a result, focused on mutations in the protease cleavage site and onset of toxicity.
- SEB In the intestinal tract, SEB is almost present in such a truncated form due to trypsin derived from ⁇ , and it is thought that SEB expresses its activity.
- human SEB antibodies include antibodies against multiple linear epitopes from analysis using Western plots, etc., indicating that SEB is processed not only as a superantigen but also as an antigen into antigen-presenting cells. It has been suggested that antigen is presented. Therefore, mutations in the protease cleavage site may alter antibody-inducing ability. It is.
- the present inventors focused on lysine at position 97 and lysine at position 98 based on the information and analysis results described above. Structurally, these two amino acids exist on the loop that connects the N-terminal domain and the C-terminal domain of SEB, and the movement and cleavage of this loop may play a significant role in the structural changes involved in the expression of SEB activity. Sex is considered. Furthermore, the present inventors have found that this site is a cleavage site for forceepsin B as well as trypsin.
- Cathepsin B is an enzyme that has been shown to be significantly involved in antigen processing in antigen presenting cells. Therefore, it was predicted that mutagenesis at this site would have a significant effect on the dynamics of SEB, antigen presentation and toxicity in vivo.
- the present inventors have used a genetic engineering technique to modify the amino acid sequence of SEB in which the asparagine at position 23 in the SEB amino acid sequence has been substituted with tyrosine to give a lysine at position 97 and a lysine at position 98.
- the mutant in which both lysines at positions 97 and 98 were substituted with serine showed good expression. .
- the notation relating to the modification used in the present specification is the notation of the amino acid name before the modification before the number at the amino acid residue position and the amino acid name after the modification after the modification.
- N Asparagine
- Y Tetyrosine
- N23YK97SK98S is The 23rd (23rd) N (Asn: asparagine) from the N-terminus is replaced by Y (Tyr: tyrosine) and the 97th (97th) and 98th (98th) (Lys: lysine) are replaced by S (Ser: Serine).
- SEQ ID NO: 2 The amino acid sequence of the N23YK97SK98S variant is shown in SEQ ID NO: 2.
- this purified variant exhibits resistance to trypsin and cathepsin B treatment, but has sufficient reactivity with the SEB antibody, and has a natural antigenicity as SEB. It was thought that there was no big difference from the type.
- the N23YK97SK98S variant had a lymphocyte proliferating activity that was reduced by a factor of about 1,000,000 compared to the native type and to about 1 / 10,000 compared to the N23Y variant.
- the modified N23YK97SK98S of the present invention also has a markedly reduced cytodynamic force-inducing stimulating activity. It showed an excellent effect of reducing the activity, such as no production induction.
- Some toxicity caused by SEB may be due to monoforces produced by leukocytes stimulated by SEB, especially tumor necrosis factor-spike (TNF-spike). Therefore, a decrease in the lymphocyte proliferation stimulating activity or the activity of inducing cytotoxicity is an indicator of a decrease in toxicity due to SEB.
- FIG. 1 shows the results of SDS-PAGE examination of the N23Y modified SEB and the N23YK97SK98S modified SEB-treated fragment of the present invention after trypsinization.
- T trypsinized
- M molecular weight standard.
- FIG. 2 Purified SEB N23Y mutant or N23YK97SK9 of the present invention was expressed in BALBZc mice.
- FIG. 7 is a graph showing the distribution of anti-SEB antibody titers in mice after inoculation of the 8S variant.
- FIG. 3 The reactivity of the antiserum obtained by immunizing the mutant N23YK97SK98S of the present invention with N2
- FIG. 4 is a graph showing the cell growth stimulating activity of the modified N23YK97SK98S of the present invention in comparison with wild-type SEB and the N23Y modified SEB.
- FIG. 5 shows that the N23YK97SK98S variant of the present invention has
- FIG. 6 shows that the modified N23YK97SK98S of the present invention has a
- FIG. 7 shows that the modified N23YK97SK98S of the present invention was orally administered to induce anti-SEB antibodies.
- the wild-type SEB (SEB prepared based on a genetic recombination technique having the same amino acid sequence as SEB derived from Staphylococcus aureus), which is the basis of the SEB variant according to the present invention, is, for example, schematically described It can be prepared by the following method.
- a 5 ′ sense primer and a 3 ′ antisense primer can be synthesized using a DNA synthesizer or the like. Plaques are collected by plaque hybridization using the primers and chromosomal DNA present in a commercially available Staphylococcus aureus DNA library, and PCR is further performed using sense primers to extract DNA from the resulting band. . Then, the extracted DNA is inserted into a suitable cloning vector and cloned.
- the cloned gene encoding SEB is digested with restriction enzymes such as Sacl, HindIII, EcoRI, BamHI, Xbal, Sail and Pstl, and the restriction enzymes Xmnl, Hindlll, EcoRI, BamHI, Xbal, Sail and Recombinant DNA is obtained by incorporation into a vector cut with Pstl et al. (Sambrook et al., Molecular Cloning, Second Edition, Chapter 9, 1989, New York, Cornoredo 'Spring' Harbor 'Laboratory' Press).
- a secretion expression vector pTrc99A or the like can be suitably used.
- a transformant By incorporating the obtained recombinant DNA into an appropriate host, for example, Escherichia coli, a transformant can be obtained. After culturing the transformant by a conventional method, cells are collected after completion of the culture, and the cells are disrupted by a conventional method to obtain a desired wild-type SEB from the suspension. In some cases, wild-type SEB is preferably secreted into the culture supernatant depending on the conditions. In this case, the culture supernatant can be used as a starting material for preparation. The starting material is purified by a purification means such as immunoaffinity chromatography in which an anti-SEB monoclonal antibody is bound to an adsorbent.
- the buffer of the final preparation used in various tests is preferably a tris-hydrochloric acid buffer, a phosphate buffer or the like.
- the modified SEB of the present invention can be prepared according to the preparation of the wild-type SEB described above. [0035] In order to maintain the prepared wild-type SEB and the modified SEB to the maximum, it is preferable that the wild-type SEB and the SEB variant be stored within 4 days when stored at 4 ° C. Alternatively, the SEB variants of the present invention can be stored in a suitable environment such as gelatin, salts, sugars, sugar alcohols or amino acids.
- the vaccine preparation of the present invention can be prepared by a known method by combining a modified SEB as an active ingredient and a known suitable excipient.
- the final dosage form of the drug product may be in the form of a powder (solid), solution, or syrup that allows subcutaneous, intramuscular, or oral administration.
- a SEB variant alone or an adjuvant typified by an aluminum adjuvant and freeze-dried into a solid form with a suitable dosage form such as a carbohydrate, a sugar, a sugar alcohol and an amino acid, or SEB
- a liquid preparation or the like in which the variant is dissolved in physiological saline and a suitable buffer having an acceptable ionic strength is a preferred embodiment.
- SEB O. lig—100 mg (0.002—2 mg / Kg body weight), preferably 1 ⁇ g—5 mg (0.02 / ig-100 / ig / Kg body weight) per administration Drugs containing variants are preferred.
- the effective dose of the vaccine preparation based on the SEB variant or a derivative thereof of the present invention varies depending on, for example, the age, symptoms and severity of the administration subject, and ultimately varies according to the intention of the physician.
- it when converted to a modified SEB, it is 0.1 ⁇ g to 1 mg per adult per day, preferably 1 ⁇ g to 5 mg in 1 or 2 divided doses. In some cases, it can be used in combination with another drug such as a steroid.
- the "derivative" of the SEB variant of the present invention refers to a SEB variant having at least one amino acid residue substitution further modified with an amino acid. Any amino acid residue in the amino acid sequence of the natural SEB other than the acid, in which the amino acid residue has been further substituted, deleted, or inserted, and which has the same activity as the modified SEB of the present invention, is the present invention. Is included.
- Nii 1 (Preparation and expression of recombinant SEB variant)
- a DNA library of Staphylococcus aureus enterotoxin A + B + D was purchased from CLONOTEC, and a plaque hybridization method was performed. Antisense synthetic DNA or PCR fragment was used as a probe. Sail cleavage sites were added to both ends of the primer to facilitate subsequent cloning operations.
- Antisense 5'-AAG TCG ACA ATA TTA GAA AAG GCA GGT ACT-3 '(SEQ ID NO: 3)
- the DNA base sequence of the obtained SEB gene was determined using an auto-sequencer.
- the SEB gene obtained as described above did not contain a mutation.
- the SEB gene that does not contain the promoter region is cut with Sail, cloned into the secretory expression vector pTrc99A (Pharmacia Biotech) having the same cleavage site, and inserted into the normal direction. Induction was performed using E. coli, and it was confirmed that SEB was secreted and expressed.
- PCR polymerase chain reaction
- PCR was performed using Taq polymerase and a DNA thermal cycler from Perkin Elmer Cetus (Norwalk, CT, USA) according to the report of Saiki et al. (Science vol. 239, p. 487 (1988)).
- 1 minute denaturation step (94 ° C) to denature and dissociate double-stranded template DNA
- 2 minute annealing step 55 ° C to associate primer with template
- 2 minute extension for synthesis The process (72 ° C) was performed for 30 35 cycles.
- the template concentration was 1 ⁇ M InM
- the oligonucleotide primer concentration was ImM.
- variants were prepared using PCR primers having mutations in the nucleotide sequence so that the amino acids at positions 97 and 98 were converted to the desired amino acids by PCR. Mutagenesis was performed as follows.
- a primer corresponding to a region corresponding to the 5 'end of SEB to which the Sfil sequence of pTrc99A / N23Y was added was used as a primer, and a primer corresponding to the 3' end of SEB to which Notl recognition sequence was added was used as a primer.
- a primer (antisense) corresponding to the corresponding region was synthesized. Furthermore, K97SK98S sense primer and antisense primer were synthesized to convert the amino acids at positions 97 and 98.
- K97SK98S antisense primer [0052] pTrc99A / N23Y was made into type III, and the 5 'region containing the mutation with the primer and the K97SK98S antisense primer was replaced with the K97SK98S sense primer and the 3' region containing the mutation with the primer. was amplified by the PCR method.
- the expression of the SEB variant was performed using the variant gene inserted into the pTrc99A vector.
- the gene-incorporated E. coli was cultured in a medium containing 4% CIRCLEGROW (BIO 101 Inc., Vista, CA, USA) and ampicillin (50 mg / ml) at 37 ° C for 18 hours, and the cells were collected. Suspend the culture medium so that the OD 550nm is 0.3-1.0, add 2mM isopropyl-B-D (-)-thiogalatatopyranoside (IPTG) and shake at 37 ° C. Induction was thus performed. After induction, Escherichia coli as a host was removed by centrifugation, followed by filtration through a 0.45 ⁇ m filtration membrane.
- the culture supernatant thus prepared was passed through a Sepharose 4B column on which anti-SEB monoclonal antibody SA58_2_6IgG had been immobilized, and the contained SEB variant was adsorbed. After washing with 0.1 M Tris HCl (pH 8.0), elution was performed with 4 M MgCl. Elution fraction, 20 volumes
- the amount of endotoxin contained in the SEB variant preparation was removed so that the final dose was SlOng / mouse or less, and the experiment was performed using males.
- the lethal toxicity of SEB after D-galactosamine administration was expressed when more than 100 ⁇ gZ mice were administered, so the dose of the modified SEB was 100 xgZ mice.
- the mortality of the mouse was higher in the native SEB and the wild-type SEB, and the lethal toxicity was shown to be lower in the other variants.
- the natural SEB means an intestinal endotoxin derived from Staphylococcus aureus
- the wild-type SEB means an SEB prepared based on a genetic recombination technique having the same amino acid sequence as the natural SEB. .
- SEB modified form (100 g / head) Mortality rate (total number of dead individuals Z total number of individuals)
- SEB-N23Y was decomposed into two fragments, N-terminal (near 21 Kda) and C-terminal (near lOKda) by trypsin treatment, whereas the modified SEB-N23YK97SK98S had the same position (32 kda ) And two bands around 28Kda.
- FIG. 1 shows that these bands had reactivity with SEB antibody (FIG. 1).
- FIG. 1 As a result of N-terminal analysis, these two bands have the same SEB N-terminal sequence, and the band around 28 Kda has reduced basicity as a result of digestion of lysine on the C-terminal side with trypsin. It was considered that the SEB-N23YK97SK98S variant migrated at a position close to the theoretical value (28.3 Kda).
- the modified SEB-N23YK97SK98S had sufficient resistance to serine proteases that could not be cleaved by trypsin at other sites.
- it exhibited the same behavior with respect to cathepsin B, and it was confirmed that it had resistance to cathepsin B.
- BALB / c mice 3-4 weeks old (female) were inoculated intraperitoneally with 20 ⁇ g of the purified SEB-N23YK97SK98S variant or purified SEB-N23Y after forming FCA and emulsion.
- the endotoxin content of the purified variant and purified SEB-N23Y was less than 0.05 EU / mg.
- FIG. 2 shows the distribution of the anti-SEB antibody titer of each mouse measured at a dilution of 10,000-fold.
- the serum anti-SEB antibody titer of the group immunized with the variant was significantly increased as compared with the SEB-N23Y immunized group.
- PBMC peripheral blood mononuclear cell
- IX 10 5 / Ueru become as 96 Uwerupureto
- SEB, SEB- N23Y, N23YK97SK98 variants were stimulated at concentrations of 0.01, 1, 100, and 10,000 ng / mL for 3 days, and 16 hours before harvest, tritium-thymidine (0.5 / i Ci) was incorporated and proliferation-inducing activity was examined. .
- PBMC peripheral blood mononuclear cell
- Each of the above PBMCs was cultured for 6 days in the presence of the same concentration of the SEB variant, and the degree of T cell blastogenesis was determined by flow cytometry (hereinafter sometimes referred to as "FACS") FSC. Investigated by / SSC analysis. As a result, N23Y induced significant blastogenesis in less than 40% of cells at Ing / mL or higher, but the inducing activity of the modified N23YK97SK98S was further reduced to about 1/2 that of N23Y (Fig. 5). .
- FACS flow cytometry
- Healthy human PBMCs were seeded at 1 ⁇ 10 6 / mL on a 24-well plate, and stimulated with 100 ng / mL of each of SEB, N23Y and a cathepsin B site variant (N23YK97SK98S variant) for 2 days, and the supernatant was collected. .
- the production of various cytokins (TNF_a, IL-1 ⁇ , IL-6, IL_8, IL-12, IFN- ⁇ , IL_lra, IL_4, IL_10, and GM-CSF) in the culture supernatant was measured using an ELISA kit (CytoSets , CytoFix, Asahi Techno Glass Co., Ltd.).
- N23YK97SK98SS female variant had significantly reduced ability to produce cytokin in comparison with N23Y and SEB.
- the production of IL-11 ⁇ , IL-6, TNF—H, IL-12, GM—CSF, and IFN— ⁇ is significantly reduced, while the IL Production of 1-4 was maintained at about the same level as ⁇ 23 ⁇ ( Figure 6).
- the modified SEB of the present invention has resistance to proteases, particularly trypsin and cathepsin B, and has extremely reduced toxicity as compared with the conventional modified SEB. Therefore, the SEB variant (N23YK97SK98S variant) of the present invention is a vaccine for preventing and treating severe diseases caused by opportunistic infections and toxins produced by bacteria resistant to antibiotics, and type 1 allergic diseases. Can be used effectively.
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Priority Applications (3)
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EP04772501A EP1661911B8 (en) | 2003-09-05 | 2004-08-31 | Protease-resistant modified seb and vaccine containing the same |
JP2005513635A JP4571586B2 (ja) | 2003-09-05 | 2004-08-31 | プロテアーゼ耐性seb改変体およびそれを含むワクチン |
US12/644,952 US7947290B2 (en) | 2003-09-05 | 2009-12-22 | Protease-resistant modified SEB and vaccine containing the same |
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JP2003314187 | 2003-09-05 | ||
JP2003-314187 | 2003-09-05 |
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US10570499 A-371-Of-International | 2004-08-31 | ||
US12/644,952 Continuation US7947290B2 (en) | 2003-09-05 | 2009-12-22 | Protease-resistant modified SEB and vaccine containing the same |
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WO2005023853A1 true WO2005023853A1 (ja) | 2005-03-17 |
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US (1) | US7947290B2 (ja) |
EP (1) | EP1661911B8 (ja) |
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Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
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KR100944034B1 (ko) | 2007-04-19 | 2010-02-24 | 한올제약주식회사 | 프로테아제-내성 폴리펩티드의 경구 투여 제형 |
US9084751B2 (en) | 2004-07-16 | 2015-07-21 | Swecure Ab | Prevention of allergy in children |
CN112028994A (zh) * | 2020-07-30 | 2020-12-04 | 北京三安新特生物科技有限公司 | 抗金黄色葡萄球菌肠毒素b的抗体、检测试纸及试剂盒 |
JP2023512344A (ja) * | 2020-12-18 | 2023-03-24 | ビオメディツィニッシュ・フォルシュング・ウント・ビオ-プロドゥクテ・アクチェンゲゼルシャフト | ブドウ球菌外毒素防御ワクチン |
Families Citing this family (2)
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ES2619943T3 (es) | 2007-01-03 | 2017-06-27 | Morphotek, Inc. | Anticuerpos de alta afinidad que neutralizan la enterotoxina B estafilocócica |
WO2023247747A1 (en) | 2022-06-23 | 2023-12-28 | Biomedizinische Forschung & Bio-Produkte AG | Protective staphylococcal exotoxin vaccine |
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JPH08500328A (ja) * | 1992-01-28 | 1996-01-16 | ナショナル ジューイッシュ センター フォア イミュノロジィ アンド レスパラトリィ メディスン | 突然変異したスーパー抗原の保護作用 |
WO1999040935A1 (fr) * | 1998-02-15 | 1999-08-19 | Juridical Foundation The Chemo-Sero-Therapeutic Research Institute | Nouveaux medicaments preventifs/curatifs de l'immunopathie |
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US5728388A (en) | 1989-10-03 | 1998-03-17 | Terman; David S. | Method of cancer treatment |
US5935568A (en) | 1995-05-18 | 1999-08-10 | National Jewish Medical & Research Center | Gene therapy for effector cell regulation |
US6248329B1 (en) * | 1998-06-01 | 2001-06-19 | Ramaswamy Chandrashekar | Parasitic helminth cuticlin nucleic acid molecules and uses thereof |
-
2004
- 2004-08-31 EP EP04772501A patent/EP1661911B8/en not_active Not-in-force
- 2004-08-31 WO PCT/JP2004/012545 patent/WO2005023853A1/ja active Search and Examination
- 2004-08-31 JP JP2005513635A patent/JP4571586B2/ja not_active Expired - Fee Related
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Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
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JPH08500328A (ja) * | 1992-01-28 | 1996-01-16 | ナショナル ジューイッシュ センター フォア イミュノロジィ アンド レスパラトリィ メディスン | 突然変異したスーパー抗原の保護作用 |
WO1999040935A1 (fr) * | 1998-02-15 | 1999-08-19 | Juridical Foundation The Chemo-Sero-Therapeutic Research Institute | Nouveaux medicaments preventifs/curatifs de l'immunopathie |
Non-Patent Citations (8)
Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US9084751B2 (en) | 2004-07-16 | 2015-07-21 | Swecure Ab | Prevention of allergy in children |
KR100944034B1 (ko) | 2007-04-19 | 2010-02-24 | 한올제약주식회사 | 프로테아제-내성 폴리펩티드의 경구 투여 제형 |
CN112028994A (zh) * | 2020-07-30 | 2020-12-04 | 北京三安新特生物科技有限公司 | 抗金黄色葡萄球菌肠毒素b的抗体、检测试纸及试剂盒 |
JP2023512344A (ja) * | 2020-12-18 | 2023-03-24 | ビオメディツィニッシュ・フォルシュング・ウント・ビオ-プロドゥクテ・アクチェンゲゼルシャフト | ブドウ球菌外毒素防御ワクチン |
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EP1661911B1 (en) | 2012-11-07 |
US20100166793A1 (en) | 2010-07-01 |
JPWO2005023853A1 (ja) | 2008-01-17 |
EP1661911B8 (en) | 2013-02-13 |
EP1661911A1 (en) | 2006-05-31 |
EP1661911A4 (en) | 2008-09-17 |
JP4571586B2 (ja) | 2010-10-27 |
US7947290B2 (en) | 2011-05-24 |
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