WO2009003425A1 - Obtención de un preparado vacunal homogéneo para el tratamiento del cáncer - Google Patents
Obtención de un preparado vacunal homogéneo para el tratamiento del cáncer Download PDFInfo
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- WO2009003425A1 WO2009003425A1 PCT/CU2008/000005 CU2008000005W WO2009003425A1 WO 2009003425 A1 WO2009003425 A1 WO 2009003425A1 CU 2008000005 W CU2008000005 W CU 2008000005W WO 2009003425 A1 WO2009003425 A1 WO 2009003425A1
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- hregf
- vaccine composition
- rp64k
- vaccine
- protein
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Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic 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
- A61K39/0005—Vertebrate antigens
- A61K39/0011—Cancer antigens
- A61K39/00113—Growth factors
-
- 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
- A61P37/02—Immunomodulators
- A61P37/04—Immunostimulants
-
- 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
- A61K2039/60—Medicinal preparations containing antigens or antibodies characteristics by the carrier linked to the antigen
- A61K2039/6031—Proteins
Definitions
- the present invention relates to biotechnology, and more specifically to the field of human health.
- the present invention describes protective and / or therapeutic vaccines for cancer, and particularly provides vaccine compositions that allow a significant increase in the immune response against Epidermal Growth Factor (EGF).
- EGF Epidermal Growth Factor
- Growth Factor whose oncological relevance has been widely demonstrated for the growth of tumors mainly of epithelial origin.
- R-EGF EGF receptor system
- R-EGF EGF receptor system
- its ligands constitute a molecular complex whose interaction specifically regulates cell growth and its impact on the uncontrolled growth of tumors of epithelial origin has been demonstrated.
- the deregulation of the paracrine and autocrine processes of activation of (R-EGF) is given both by the overproduction of growth factors and by the high synthesis and / or the mutation of its receptors.
- EGF is a 53 amino acid polypeptide, with an apparent molecular weight of 6045 Da. This polypeptide was isolated and purified for the first time by Cohen S., J. Biol. Chem. (1962) 237, 1,555).
- the EGF is part of the R-EGF family of ligands, this family is constituted by structurally and functionally related proteins.
- the other members of this family are: TGF, anfiregulin (AR), cryptol (CR1), heparin-binding growth factor, betacellulin, epiregulin.
- the family of poxviruses includes proteins related to EGF, among them the most characterized is the growth factor of vaccinia virus (VGF).
- R-EGF receptor-binding protein
- R-EGF is a glycoprotein of approximately 170 kDa, whose gene has been cloned and sequenced.
- the intracellular domain of this receptor is associated with the activity of tyrosine kinase proteins that show structural homology with the product of the oncogene v-erb-B, which shows a relationship with the process of malignant transformation (Heldin CH. (1984), CeII 37,
- EP 0 657 175 describes a vaccine composition containing autologous EGF coupled to a transporter protein and said complex inhibits the growth of EGF-dependent tumors through an auto immune effect.
- the vaccine composition described in EP 0 657 175, are conjugate vaccines containing EGF, coupled to a transporter protein (rP64k protein, Cholera toxin B chain, tetanus toxoid protein and / or monoclonal antibodies) and as a consequence a heterogeneous and poorly reproducible mixture of conjugated species is obtained.
- a transporter protein rP64k protein, Cholera toxin B chain, tetanus toxoid protein and / or monoclonal antibodies
- the novel vaccine preparation described in the present invention comprises as an active principle the autologous EGF and is characterized by having a homogeneous chemical composition and defined purity, an enhanced immunogenicity, a substantially increased clinical activity and significantly decreases the number of immunizations for its therapeutic action
- the decrease in the content of the autologous antigen in the vaccine preparation of the present invention surprisingly does not decrease, but causes significant increases in the titers of anti-EGF antibodies in humans.
- the decrease of the content of EGF and its derivatives in the vaccine composition was obtained through the development of a method of membrane purification, which is also part of this invention.
- the present invention also provides a homogeneous and reproducible vaccine preparation that allows to increase the clinical effect thereof by decreasing the number of immunizations, which provides a great advantage in cancer therapy and for the patient, in comparison with previously described vaccine compositions.
- the vaccine composition according to the present invention can be used in conjunction with suitable adjuvants such as aluminum hydroxide or Montanide
- the present invention involves a sanitary procedure for obtaining the appropriate vaccine preparation for administration to patients parenterally.
- the method comprises a suitable method of covalent conjugation of the hrEGF, to the rP64k transporter protein and the purification thereof using Ultrafiltration membranes in a range of 50-10OkDa. to remove biologically inactive conjugate species (lacking immunogenic activity) and other chemical impurities.
- the present invention provides a novel vaccine preparation based on autologous EGF, characterized by having a homogeneous chemical composition and defined purity, enhanced immunogenicity and substantially increased clinical activity. Additionally, the vaccine presentation described in the present invention offers the possibility of significantly decreasing the number of administrations necessary to the patient, when it is required to increase the therapeutic dose, due to the fact that this vaccine composition is obtained from a methodology that allows to achieve different concentrations per milliliter of the hrEGF-rP64k conjugate species, known as potency
- This new vaccine preparation is obtained from the surprising result that the decrease in the content of the autologous antigen in the vaccine composition through a novel purification process, does not decrease, but it allows to significantly increase the immune response against the EGF itself compared to heterogeneous compositions used as vaccines based on the EGF.
- the vaccine preparation obtained from the decrease of the autologous antigen content through the ultrafiltration membrane purification methodology has a homogeneous composition where immunologically active species (hrEGF-rP64k conjugate species) show molar masses greater than 6OkDa .
- This Vaccine preparation shows a chromatographic separation profile by molecular exclusion characterized in that where the species that carry therapeutic activity of this vaccine preparation, represent more than 90% of the total area of the chromatogram corresponding to the vaccine preparation as a whole, as shown in Figure 4
- This vaccine preparation is further characterized by a chemical conjugation relationship between the hrEGF and rP64k proteins of (1: 2), this means that there are 2 conjugated hrEGF molecules for each rP64k molecule Methodology for obtaining the vaccine preparation
- a method is developed to decrease the content of hrEGF in the heterogeneous mixture of conjugates; enriching the vaccine preparation in conjugated species of high molecular weight (immunologically active species) and glutaraldehyde free.
- the purification procedure using ultrafiltration membranes in a range between 50-10OkDa developed and described in the present invention consists of two stages: In the initial stage successive changes of buffer solution (diafiltration) are made to remove the glutaraldehyde and remove the excess of autologous protein, either free or forming hrEGF-hrEGF conjugates of different sizes.
- the second stage is the concentration of the purified chemical conjugate, in which more than 90% of its composition corresponds to the immunogenic species hrEGF-rP64k.
- the concentration stage the initial volume of the chemical conjugate is reduced to a final protein concentration (hrEGF-rP64k conjugate species) in a range between 1-12 mg / mL.
- hrEGF-rP64k conjugate species The uniqueness of this methodology ensures that the vaccine preparation obtained has a homogeneous composition and defined purity, characterized by the majority presence of immunologically active species: hrEGF-rP64k conjugate species.
- hrEGF polymeric autologous protein
- the reduction of the autologous protein content in the vaccine preparation causes in patients increases of at least 2 times the maximum anti-EGF antibody titer.
- the procedure described in the present invention to guarantee an adequate chemical conjugation between both proteins consists of a single step and begins by mixing the previously concentrated hrEGF protein (> 6 mg / mL) and the rP64k protein (> 1 mg / mL) in the conjugation reactor. To this protein mixture is subsequently added the PBS / MgCl 2 solution (pH 6.8-7.2) and the 0.5% glutaraldehyde conjugation solution. The mixture is kept under constant stirring for 2 hours at a temperature 22 ° C ⁇ 2 ° C. The final protein concentration in the reaction mixture for hrEGF is 0.82 mg / mL and for rP64k protein it is 0.89 mg / mL.
- the concentration of total proteins during the conjugation reaction is 2 mg / mL and the final concentration of glutaraldehyde in the reaction mixture is 0.05%.
- the vaccine composition according to the present invention can be used in conjunction with suitable adjuvants, such as aluminum hydroxide or Montanide.
- the present invention involves a sanitary procedure for obtaining the vaccine preparation, administered parenterally that minimizes the opportunities for microbial contamination of the vaccine preparation during its obtaining, allows the chemical conjugate purification methodology to be performed in a few hours and facilitates the increase in the volume of the vaccine preparation to obtain among other advantages.
- Example 1 describes the molecular characterization of the vaccine preparation described in the present invention.
- Molecular characterization includes: the HPLC-FG chromatographic identification of the conjugate species formed during the conjugation reaction, the determination of the conjugation ratio between the hrEGF and rP64k proteins and the definition of the peptide map that characterizes this novel vaccine preparation .
- Example 2 describes the mouse bioassays performed to evaluate the immunogenicity of the novel vaccine preparation described in the present invention.
- Example 3 describes the efficacy of the clinical use of the present vaccine preparation in the treatment of lung tumors of epithelial origin.
- Example 4 describes the obtaining of vaccine preparations adjusted to different powers (total milligrams of hrEGF-rP64k / vial conjugate), using the same conjugation and purification methodology described in the present invention.
- Example 5 shows the removal of glutaraldehyde used in the conjugation reaction, by means of the methodology of purification of the chemical conjugate through a 5OkDa ultrafiltration membrane.
- EXAMPLE 1 Molecular characterization of the vaccine preparation described in the present invention (Vaccine Preparation A).
- hrEGF-hrEGF and rP64k-rP64k were used as conjugated homoligomeric indicators.
- the chromatographic profiles shown by the homoligomeric conjugates (hrEGF-hrEGF and rP64k-rP64k) were compared with the chromatographic profile shown by the chemical conjugate obtained from the conjugation reaction between the unpurified hrEGF and rP64k proteins.
- Figure 1 shows the profile of the homoligomeric conjugate rP64k-rP64k and the profile that is obtained when performing the chemical conjugation between the hrEGF and rP64k proteins (chemical conjugate without purification by ultrafiltration).
- FIG. 1 shows the results obtained after performing a denaturing electrophoresis under conditions of reduction and Western Blot of the vaccine preparation purified by ultrafiltration. The development was performed with an anti-EGF antibody conjugated with alkaline phosphatase.
- the EGF is a protein of apparent molecular weight of 6kDa therefore it must be located in the lower area of the gel.
- the moles present in each of its constituent proteins were determined.
- a vaccine preparation obtained from a 50 kDa ultrafiltration membrane purification process was taken and the chromatographic fraction corresponding to the hrEGF-rP64k conjugate species was collected. These fractions were subjected to amino acid analysis.
- the determination of the conjugation ratio was performed by 2 methods. 1. A method was based on the estimation of the amount of Phenylalanine (Phe) and Threonine (Thr) in the vaccine preparation (amino acids only present in the rP64k molecule). The amounts of these amino acids were used to determine the amount of rP64k.
- the amount of EGF was determined. 2.
- the other method was based on the direct determination of the relative amounts of each protein, using the amino acid sequence and based on the quantification of the amino acids Asparagine + Aspartic (Asx), Glutamine + Glutamic (GIx), Glycine (GIy) and Alanine (Ala) (residues highly stable to acid hydrolysis and which are commonly used in protein quantification). The results obtained by each method are shown in Tables 1 and 2 below.
- Table 1 Determination of the ratio of conjugation between the hrEGF and rP64k proteins by performing the determination of amino acid composition of the vaccine preparation purified by ultrafiltration, based on the method of determining the amount of phenylalanine (Phe) and Threonine (Thr)
- Table 2 Determination of the ratio of conjugation between the hrEGF and rP64k proteins by determining the amino acid composition of the vaccine preparation purified by ultrafiltration, based on the method of determining the relative amounts of the amino acids Asparagine + Aspartic (Asx), Glutamine + Glutamic (GIx), Glycine (GIy) and Alanine (Ala)
- Figure 5 shows the maps of peptides obtained from two vaccine preparations obtained independently, where great similarity is observed in the peptides that appear and the existing proportion of each of them.
- the reproducibility between the peptide maps obtained from both vaccine preparations is an indication of the control that exists over the conditions under which the chemical conjugation and purification procedures also described in the present invention occur.
- the inoculation of 10 mice was performed with a single dose of 200 ⁇ L of a water-in-oil emulsion (50/50% v / v) of the immunogens (conventional vaccine preparation obtained by dialysis, vaccine preparation obtained by membrane purification ultrafiltration and the permeate obtained from the UF / DF purification process, where the hrEGF-hrEGF conjugated species are found) with the Montanide ISA 51 adjuvant.
- the protein dose applied for each immunogen is referred to in Table 4.
- the extraction of the serum from the animals and the evaluation of the anti-EGF antibody titer by means of an ELISA was performed.
- the test positivity criterion was that the optical density measured at 405 nm was more than twice the average obtained for the plate blank. Plaque blank is obtained when the blocking buffer is added to the well instead of an anti-EGF serum sample.
- the Mann-Whitney test applied to the analysis of the data was performed using a statistical program.
- Table 4 Immunogens evaluated and the concentration of proteins to be inoculated by animal as appropriate for the biological activity test.
- Figure 6 shows the results of the biological activity test as evidence of the immunogenic activity of each immunogen evaluated.
- the Vaccine Preparation A produces an increase in the concentration of anti EGF antibodies (at 1/1000 dilution) 10 times higher compared to that achieved by immunizing with the conjugate corresponding to the free hrEGF and hrEGF 'polymers.
- the vaccine preparation described in the present invention produced a 2-fold increase in the concentration of anti EGF antibodies
- Point Estimate for ETA1-ETA2 is -0.2590
- EXAMPLE 3 Evaluation of the efficacy of the clinical use of the vaccine preparation described in the present invention in the treatment of lung tumors of epithelial origin.
- the objective of this example is to evaluate in patients the immunogenicity produced by the Vaccine Preparation A in comparison with the immunogenicity achieved by the Vaccine Preparation B obtained by the method of dialysis purification.
- patients carrying non-cell lung tumors were treated. Small in advanced stages.
- Two groups of patients (patients vaccinated with Vaccine Preparation B and patients vaccinated with Vaccine Preparation A, described in the present invention were defined for the study.
- the dose of vaccine preparation applied per patient for each group was the following 'vaccinated patients With Vaccine B Preparation they received 1.2 mg of total protein per immunization site, patients vaccinated with Vaccine A Preparation received 0.6 mg of total protein per immunization site, for both groups the immunization sites were 4. All patients received therapy oncospecific, at least 4 weeks before Start the essay. Immunizations were performed intramuscularly, and they continued to happen once a month.
- the humoral immune response of both groups of patients was evaluated through the obtaining of blood serum.
- the fundamental outcome variable was the specific antibody titer against EGF. This parameter was determined by means of an ELISA system. The positive values were those optical density readings that were 2 times above the value of the negative control. The antibody titer reported means the greatest dilution of positive sera.
- the analysis of the immune response for the members of each group represented in Table 6 showed that the geometric mean of the maximum antibody titer for the group of patients immunized with Vaccine Preparation A corresponds to 1: 56421, while for the group of patients immunized with Vaccine Preparation B the geometric mean of the maximum antibody titer was 1: 23515. This means that the vaccine preparation described in the present invention produced a 2-fold increase in the geometric mean of the maximum anti-EGF antibody titer in relation to that achieved with the vaccine preparation B.
- Vaccine Preparation A causes an increase of at least 100 times the titer of anti-EGF antibodies in immunized subjects
- Test 1 At the conclusion of the diafiltration, the purified chemical conjugate was concentrated to a final protein concentration of 2 mg / mL.
- Test 2 At the conclusion of the diafiltration, the purified chemical conjugate was concentrated to a final protein concentration of 5 mg / mL.
- Test 3 At the conclusion of the diafiltration, the purified chemical conjugate was concentrated to a final protein concentration of 12 mg / mL.
- Table 7 Values corresponding to the percentage of homogeneity in vaccine preparations adjusted to different final protein concentrations
- the residual glutaraldehyde content in the Vaccine Preparation B and in the Vaccine Preparation A was quantified, by a reverse phase chromatographic separation method (HPLC-RP) using a C-8 column. This method required first, the precipitation of the protein contained in the samples with perchloric acid and then a reaction with phenylhydrazine.
- Figure 7 shows that the residual content of this impurity present in both vaccine preparations was less than 0.2 ug / mL or less than 0.002 ppm, (upper limit concentration established for this impurity) starting from an initial concentration of 530 mg / mL or 530 ppm Although both preparations comply with the specification established for this impurity, Figure 7 illustrates that the vaccine preparation A obtained from applying the method of purification of the chemical conjugate by Ultrafiltration membranes, has a lower content of glutaraldehyde, which provides great safety. for the patient to said vaccine preparation
- Figure 1 The superposition of the chromatogram of a homoligomeric conjugate rP64k-rP64k and the chromatogram of a chemical conjugate EGF-P64k without purification is shown. Encased in the circle shows the coincidence of the profiles between both chromatograms.
- the X axis presents the time at which each component of the sample elutes and the Y axis represents the intensity value at 216 nm of each component of the sample, as an indicator of the protein concentration in the same.
- FIG. 1 Immunodetection with an anti EGF antibody conjugated to alkaline phosphatase after electrophoretic separation in SDS-PAGE for vaccine preparation obtained by ultrafiltration.
- Line 1 Molecular weight marker
- Line 2 Vaccine preparation Figure 3.
- the superposition of the chromatogram corresponding to the hrEGF-hrEGF homoligomeric conjugate and the chromatogram of an unpurified hrEGF-rP64k chemical conjugate is shown. Encased in the circle shows the coincidence of species between both chromatograms. .
- the X axis presents the time at which each component of the sample elutes and the Y axis represents the intensity value at 216 nm of each component in the sample, as an indicator of the protein concentration in the same.
- FIG. 4 Chromatographic profile corresponding to the vaccine preparation described in the present invention.
- the shaded area corresponds to the hrEGF-rP64k conjugate species.
- the X axis presents the time at which each fraction elutes and the Y axis represents the intensity value at 216 nm of each component in the sample, as an indicator of the protein concentration in it.
- FIG. 5 Peptide map of the chromatographic fraction corresponding to the hrEGF-rP64k conjugate. The results obtained for two independently purified vaccine preparations are presented through an ultrafiltration membrane. The axis of the X represents the time and the axis of the Y represents the units of milli intensity.
- Figure 6 Titles of anti EGF antibodies in mice immunized with various immunogens: conventional vaccine preparation, vaccine preparation purified by ultrafiltration and permeate from the ultrafiltration purification method of the vaccine preparation The X axis corresponds to the dilutions made to the serum obtained from each animal and the Y axis represents the 405 nm optical density value of each sample as an indicator of the protein concentration in it.
- FIG. 7 HPLC-RP chromatographic profile obtained during the quantification of the residual glutaraldehyde content for the vaccine preparation obtained from the purification method by using the 5OkDa ultrafiltration membranes (Vaccine Preparation A) and for the vaccine preparation obtained using the method of dialysis membrane purification (Vaccine Prepared B).
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Abstract
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JP2010513634A JP5368437B2 (ja) | 2007-06-29 | 2008-06-26 | 癌治療のための均一なワクチン製剤の製造方法 |
EP08757902A EP2184072B1 (en) | 2007-06-29 | 2008-06-26 | Production of an homogeneous vaccine preparation for cancer treatment |
AT08757902T ATE536885T1 (de) | 2007-06-29 | 2008-06-26 | Produktion einer homogenen impfstoffzubereitung zur behandlung von krebs |
NZ582829A NZ582829A (en) | 2007-06-29 | 2008-06-26 | A VACCINE COMPOSITION COMPRISING hrEGF-rP64K, WHEREIN EACH P64K IS BOUND TO TWO hrEGF (EPIDERMAL GROWTH FACTOR) MOLECULES |
DK08757902.5T DK2184072T3 (da) | 2007-06-29 | 2008-06-26 | Produktion af et homogent vaccinepræparat til cancerbehandling |
CA2692340A CA2692340C (en) | 2007-06-29 | 2008-06-26 | Production of an homogeneous vaccine preparation for cancer treatment |
BRPI0813662A BRPI0813662B8 (pt) | 2007-06-29 | 2008-06-26 | composição vacinal, processo para a obtenção de uma composição vacinal, e, uso de uma composição vacinal |
PL08757902T PL2184072T3 (pl) | 2007-06-29 | 2008-06-26 | Wytwarzanie preparatu jednorodnej szczepionki do leczenia raka |
CN200880021856XA CN101790383B (zh) | 2007-06-29 | 2008-06-26 | 用于肿瘤治疗的均质疫苗制剂的获得 |
EA201070080A EA021905B1 (ru) | 2007-06-29 | 2008-06-26 | Однородная вакцинная композиция для лечения опухоли и способ её получения |
PE2013000227A PE20131037A1 (es) | 2007-06-29 | 2008-06-26 | COMPOSICION INMUNOGENICA QUE COMPRENDE UN CONJUGADO hrEGF-rP64k |
SI200830563T SI2184072T1 (sl) | 2007-06-29 | 2008-06-26 | Proizvodnja homogenega vakcinskega pripravka za zdravljenje raka |
RS20120101A RS52196B (en) | 2007-06-29 | 2008-06-26 | MANUFACTURE OF A HOMOGENEOUS CANCER PREPARATION FOR CANCER |
ES08757902T ES2379130T3 (es) | 2007-06-29 | 2008-06-26 | Producción de una preparación de vacuna homogénea para el tratamiento del cáncer |
AU2008271798A AU2008271798B2 (en) | 2007-06-29 | 2008-06-26 | Production of an homogeneous vaccine preparation for cancer treatment |
US12/664,545 US8778879B2 (en) | 2007-06-29 | 2008-06-26 | Homogeneous vaccine composition comprising a conjugate of EGF and P64K for the treatment of tumors |
HK11100165.6A HK1145979A1 (en) | 2007-06-29 | 2011-01-10 | Production of an homogeneous vaccine preparation for cancer treatment |
US13/346,831 US8841251B2 (en) | 2007-06-29 | 2012-01-10 | Method for making a homogenous vaccine composition comprising a conjugate of EGF and 64K for the treatment of tumors |
HR20120218T HRP20120218T1 (hr) | 2007-06-29 | 2012-03-07 | Proizvodnja homogenog preparata cjepiva za liječenje raka |
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CU20070154A CU23652A1 (es) | 2007-06-29 | 2007-06-29 | Composición vacunal homogénea para el tratamiento del cáncer y su método de obtención |
CUCU154-2007 | 2007-06-29 |
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US12/664,545 A-371-Of-International US8778879B2 (en) | 2007-06-29 | 2008-06-26 | Homogeneous vaccine composition comprising a conjugate of EGF and P64K for the treatment of tumors |
US13/346,831 Division US8841251B2 (en) | 2007-06-29 | 2012-01-10 | Method for making a homogenous vaccine composition comprising a conjugate of EGF and 64K for the treatment of tumors |
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EP (1) | EP2184072B1 (es) |
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CN114594257B (zh) * | 2022-05-09 | 2022-08-05 | 北京生物制品研究所有限责任公司 | 含CpG ODN的吸附型疫苗的解吸附组合物及其应用 |
Citations (2)
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EP0657175A2 (en) | 1993-12-09 | 1995-06-14 | Centro de Inmunologia Molecular | Vaccine comprising autologous epidermal growth factor and use thereof |
WO2002045747A1 (es) * | 2000-12-08 | 2002-06-13 | Centro De Inmunologia Molecular | Combinaciones inmunoterapeuticas para el tratamiento de tumores |
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CA2261433A1 (en) * | 1993-12-09 | 1995-06-10 | Belinda Sanchez Ramirez | Composition comprising autologous epidermal growth factor |
CU23077A1 (es) * | 2000-12-06 | 2005-08-17 | Centro Inmunologia Molecular | Composicion vacunal que contiene factor de crecimiento transformante (tgf-alfa). su uso en la terapia de enfermedades malignas |
CU22999A1 (es) * | 2001-12-04 | 2004-10-12 | Centro Inmunologia Molecular | Método de tratamiento de enfermedades malignas e infecciosas crónicas |
GB0109297D0 (en) * | 2001-04-12 | 2001-05-30 | Glaxosmithkline Biolog Sa | Vaccine |
FR2844514B1 (fr) * | 2002-09-16 | 2007-10-19 | Neovacs | Produit immunogene stable comprenant des heterocomplexes antigeniques, compositions les contenant et procede de preparation |
GB0405787D0 (en) * | 2004-03-15 | 2004-04-21 | Chiron Srl | Low dose vaccines |
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