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WO1995000540A1 - Vecteur et immunogene de synthese - Google Patents

Vecteur et immunogene de synthese Download PDF

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Publication number
WO1995000540A1
WO1995000540A1 PCT/US1994/005981 US9405981W WO9500540A1 WO 1995000540 A1 WO1995000540 A1 WO 1995000540A1 US 9405981 W US9405981 W US 9405981W WO 9500540 A1 WO9500540 A1 WO 9500540A1
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WIPO (PCT)
Prior art keywords
carrier
amino acid
peptide
biologically active
pair
Prior art date
Application number
PCT/US1994/005981
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English (en)
Inventor
Robert Webber
Original Assignee
Robert Webber
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Robert Webber filed Critical Robert Webber
Publication of WO1995000540A1 publication Critical patent/WO1995000540A1/fr

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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
    • C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
    • 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
    • A61K39/385—Haptens or antigens, bound to carriers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/62—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being a protein, peptide or polyamino acid
    • A61K47/64—Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent
    • A61K47/641—Branched, dendritic or hypercomb peptides
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/001—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof by chemical synthesis
    • 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
    • A61K2039/64—Medicinal preparations containing antigens or antibodies characterised by the architecture of the carrier-antigen complex, e.g. repetition of carrier-antigen units
    • A61K2039/645—Dendrimers; Multiple antigen peptides
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2740/00—Reverse transcribing RNA viruses
    • C12N2740/00011—Details
    • C12N2740/10011—Retroviridae
    • C12N2740/16011—Human Immunodeficiency Virus, HIV
    • C12N2740/16111—Human Immunodeficiency Virus, HIV concerning HIV env
    • C12N2740/16122—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes

Definitions

  • the present invention relates to a novel synthetic carrier, used for constructing a biologically active component delivery vehicle such as an immunogen utilizing the same.
  • Vaccines have been developed to inoculate animals against virulent diseases such as poliomyelitis, measles, small pox, and the like. Such immunization has been effected by utilizing an attenuated or killed virus in the animal to elicit antibodies which kill the disease organism. Unfortunately, attenuated viruses have an uncertain level of danger in the animal injected with the same. In addition, the vaccines utilizing killed or attenuated viruses are easily contaminated and require special handling such as refrigeration prior to use.
  • Synthetic vaccines have attracted great interest since they do not employ killed or attenuated viruses. Rather, peptides are used to mimic portions of a particular virus which elicit the antibodies. This effect was described by Richard A. Lerner in an article entitled “Synthetic Vaccines", Scientific American. Pages 66-74, February 1983.
  • low molecular weight compounds such as peptides, nucleotides, steroids, toxins, carbohydrates, and other small organic compounds for synthetic immunogens. These low molecular weight compounds have either been isolated from natural sources or produced synthetically. In addition, these low molecular weight compounds have been attached to a carrier molecule such as a protein. Specifically, the carriers used have been keyhole limpet hemocyanin (KLH) , bovine serum albumin (BSA) , ovalbumin, bovine thyroglobulin, and the like. Such low molecular weight compound-protein conjugates are required to elicit an antibody response in an animal. It is well documented that small molecules are not recognized by the immune system.
  • KLH keyhole limpet hemocyanin
  • BSA bovine serum albumin
  • ovalbumin ovalbumin
  • bovine thyroglobulin and the like.
  • FCA Freund's complete adjuvant
  • MAP Multiple Antigenic Peptide
  • a 15 amino acid long peptide with a molecular weight of approximately 1,600 Daltons when built on a 16 branched chain core of Lys 8 -Lys 4 -Lys 2 -Lys-X-X-Resin would have a molecular weight of about 27,000 Daltons.
  • the MAP system Although useful as an antigen for the production of antibodies specific to the synthetic peptide, the MAP system only performs this function. In another words, the MAP approach eliminates the need for a high molecular weight carrier protein, but does not deliver additional biologically active components such as adjuvants, cytotoxins, fatty acids, and the like. In addition, the MAP approach suffers from numerous problems.
  • the peptide antigens are synthesized directly on the branched chain core. This does not allow for purification of the synthetic peptide.
  • the peptide antigen component of the MAP is actually a complex mixture of numerous peptides of unknown and undefined sequence.
  • various investigators have reported on the elicitation of a strong antibody response to minor impurities in preparations of synthetic peptides. It has been concluded that the only way to avoid this problem is to used highly purified synthetic peptides of known structure. MAP is incapable of solving this problem.
  • MAP does not allow for the construction of antigens with other non-peptide components such as carbohydrates, glycopeptides, glycoproteins, nucleotides, nucleopeptides, peptides, or proteins built by other methods such as solution phase synthesis or by genetic engineering.
  • structurally complex peptides are not capable of being synthesized as a MAP structure. For example, peptides with disulfide bonds, which are the best analogues of whole protein loop structures, lie in this category.
  • United States Patent 5,198,531 to Webber et al describes a notable advance in the preparation of peptides utilizing a polymeric resin.
  • the resin of the 5,198,531 patent permits the building of peptides of exceptional purity, greatly eliminating undesirable side -reactions during the process.
  • a synthetic carrier for delivering biologically active components such as antigens, cytotoxins, drugs, adjuvants, fatty acids, alone and/or in combination, at a specific level or ratio, would be a major advance in the medical and biological fields.
  • a novel and useful carrier for biologically active components to an organism is herein provided.
  • the present invention comprises a synthetic branched chain carrier for delivering at least one biologically active component to an organism.
  • the carrier employs a first unbranched peptide chain which may or may not be linked to a solid support resin.
  • the first peptide may be formed of an unbranched peptide segment or analogue of any length and include amino acids therealong that are capable of linking biologically active components, to their individual sidechains.
  • the unbranched peptide segment terminates in an amino acid having a pair of end sites, thus providing a branched site.
  • the second peptide is attached to one of the pair of end sites of the first peptide, while a third peptide is attached to the other of said pair of end sites of the first peptide.
  • the second and third peptides may be identical and each include a terminal amino acid having a pair of end sites capable of bonding a pair of biologically active components. Bonding is used to include covalent and ionic bonding, electrostatic interaction, hydrophobic interaction, Vander Waals force attraction, and the like. Thus, a quartet of end sites are available in the basic synthetic carrier of the present invention, as well as a pre-determined number of intermediate sites found along the unbranched segments of the first, second, and/or third peptide, at individual side chains.
  • the biologically active components may take the form of immunogenic peptides, adjuvants, fatty acids or alcohols, cytotoxins, drugs, nucleotides and the like. Those components may be placed at the end sites or intermediate sites in specific numbers and, thus, in specific ratios when more than one component is employed. In this regard, a plurality of components may be used in the carrier of the present invention, each having a specific biological function.
  • the biologically active component may be a receptor peptide coupled with a drug, cytotoxin, or the like.
  • the carrier of the present invention may be expanded to multiple levels of peptide segments, having branched end sites, connected to the pair of end sites of each amino acid terminating each peptide. That is to say, at each successive branched level, the number of end sites would double from the prior level. Thus, at the fifth level, utilizing the first peptide as the (zero) level, (32) end sites would be available for combining biologically reactive components.
  • a particular use of the carrier of the present invention is to form an immunogen which will elicit an immunological response in an animal.
  • Such immunogen may be considered as a vaccine in certain cases when the response concerns viral entities.
  • Another object of the present invention is to provide a synthetic carrier which is capable of delivering a multiplicity of biologically active components in pre ⁇ defined ratios.
  • a further object of the present invention is to provide a synthetic carrier for delivering at least one biologically active component to an organism which may be an immunogen.
  • Yet another object of the present invention is to provide a synthetic carrier for delivering biologically active components to achieve a variety of results such as drug treatment, delivering a cytotoxin to a specific site, and the like.
  • a further object of the present invention is to provide a synthetic carrier for delivering a biologically active component which activates major histocompatibility complexes I, II, and/or III.
  • Yet another object of the present is to provide a synthetic carrier which is useful in the production of a synthetic vaccine.
  • Fig. 1 is a depiction, of the overall matrix of the carrier of the present invention taken from levels “0" through “5", still attached to a solid support resin on which it was synthesized.
  • Fig. 2 is a formulation of a component bound to an intermediate site of the matrix of Fig. 1.
  • Fig. 3 is a depiction of MDP bound to ornithine at an intermediate site of the matrix of Fig. 1.
  • Fig 4 is a depiction of a fatty alcohol bound to a site of the matrix of Fig. 1.
  • Fig. 5 is a depiction of a fatty acid bound to a site of the matrix of Fig. 1.
  • the synthetic branched chain carrier peptide of the present invention may take the following general formulation as attached to the solid phase support resin: Formula 1
  • Lys represents a dibasic amino acid such as lysine, ornithine, 2, 3-diaminopropionic acid, 2, 4- diaminobutyric acid and the like. Such dibasic amino acids can serve as branch points and attachment sites. With reference to Fig. 1, it may be observed that a branched carrier may be built under the present invention to extend to various levels. Fig.
  • FIG. 1 shows levels 0-5 where the fifth level furnishes (32) end attachment sites capable of attaching biologically active components such as an antigen, cytotoxin, drug, monoclonal antibody, and the like.
  • the carrier depicted in Fig. 1 and described above may be built following the procedures described in United States Patent 5,198,531.
  • Fig. 2 represents a typical functional group or component designated “Com” attached to intermediate functional group site 12, denoted VW at level 2 of Fig. 1.
  • “Com” is possible in the present invention by designing in appropriate functional groups prior to the provision of the branched end chain 14.
  • Functional groups at site 12 can take the form of trifunctional amino acids like aspartic acid or glutamic acid, both of which have a - COOH functional group on their respective side chains.
  • cysteine may be employed to provide a -SH functional group on its side chain.
  • lysine may be employed in this regard utilizing its epsilon-NH 2 group. It should be noted that each of these functional groups may be differentially protected.
  • other groups may be used to incorporate more specialized components.
  • cysteine may function as an attachment site for more than one type of biologically active molecule to be delivered to an organism.
  • Fig. 3 depicts the attachment of muramyl dipeptide (MDP) to ornithine.
  • MDP is an adjuvant peptide which would be attached through routine chemistry, known in the peptide building art.
  • biological active components may be placed at these sites in the carrier of the present invention in specific numbers and in various coupling ratios. It is also practical to provide more complex biologically active components for specialized applications. Specifically, three or more components may be added to the carrier of Fig. 1 at the (32) available end sites of the fifth level, and at intermediate level sites 12 and 16. As an example, a kidney receptor peptide may be placed on the (32) available end sites at the fifth level while a drug may be attached to the intermediate site 12.
  • a tumor receptor peptide may be placed at the (32) available end sites while a cytotoxin is placed at intermediate site 12 or 16. It should be stressed that the position of the biologically active components may be reversed i.e. a drug may be placed on the (32) available end sites while a receptor peptide may be placed on any of the intermediate sites, or built into the carrier as the initial unbranched peptide or peptide analogue segment.
  • a fatty alcohol may be attached to the carboxyl terminal amino acid after it has been cleaved from the resin. This may be accomplished by addition of the fatty alcohol to the free
  • Fig. 5 illustrates that a fatty acid may be attached to the trifunctional group designated Lys after cleavage of the carrier peptide at group "PPP", from the resin. This can be achieved by protecting the epsilon -NH 2 group of Lys and treating the same with TFA.
  • the fatty acid would normally be pre-activated, with hydroxybenzotriazole and diisopropylcarbodiimide (HOBt+DIPCDI) , for example. Attachment of the fatty alcohol or fatty acid as noted in Fig. 4 and 5 may be accomplished by other known chemical processes.
  • the carrier of the present invention to bond a fatty acid or fatty alcohol as an anchor for the carrier of the present invention to liposomes, micelles, or other hydrophobic structures.
  • the fatty acid or alcohol anchor may be combined with a targeting receptor peptide and/or a drug attached to either the (32) end sites of Fig. 1 or an intermediate site such as site 12 or 16 of Fig. 1.
  • a trio of components may be placed on the carrier of the present invention in specific ratios.
  • the branched configuration of Fig. 1 may be lengthened or shortened such that the carrier includes sites at the first level only or includes sites beyond the fifth level.
  • the carrier of the present invention may be used for other purposes than to build a synthetic immunogen, this particular application is especially useful. Specifically, disulfide cyclized peptide analogues of the CD binding site of the HIV-1 virus may be synthesized and used in conjunction with the carrier of the present invention. Other synthetic immunogens may be constructed using other peptide analogues to various viral components..-
  • a first peptide (identified as "PC-1580”) was synthesized at the 0.05 mmole scale by standard Fmoc-Solid Phase Peptide Synthesis (Fmoc-SPPS ) using hydroxybenzotriazole/diisopropylcarbodiimide (HOBt/DIPCDI) procedures and a four molar excess of reagents on a Dasrin- 3 resin available from Research and Diagnostic Antibodies, Berkeley, California.
  • the sequence of the (40) amino acid long peptide was as follows:
  • the peptide was cleaved from the resin and the side chains of the trifunctional amino acids were simultaneously deprotected using 40 ml of 50% trifluoroacetic acid (TFA) in dichloromethane (DMC) containing 1ml H 2 0, 0.5 ml thioanisole, 210 mg dithiothreitol (DTT) and 38 1 anisole.
  • TFA trifluoroacetic acid
  • DMC dichloromethane
  • DTT dithiothreitol
  • the spent resin was washed with 100% acetic acid (HOAc) , 50% HOAc/50% water, and water.
  • the peptide was air oxidized for three days with constant slow mixing to form the intrachain disulfide bond and cyclize the peptide.
  • the loss of free - SH groups was monitored during the air oxidation by the quantitative DTNB assay.
  • the solution was centrifuged to remove precipitated material, and pumped at 3 ml/min onto a semi-preparative C 18 reverse phase HPLC column.
  • the peptide was eluted from the column with a gradient of acetonitrile.
  • the eluent from the column was monitored for OD at 286nm and fractions were collected. The fractions which contained peptide were transferred to pre-weighed vials, lyophilized, and analyzed.
  • EXAMPLE 2 A second peptide (identified as PC-1581) was synthesized by Fmoc-SPPS using (4) molar excess of reagents, cleaved, isolated, cyclized, purified, and analyzed by the same procedures as described above for the peptide, (PC-1580) , of Example 1.
  • EXAMPLE 3 A branched chain synthetic carrier peptide of the type depicted in Fig. 1 was synthesized and conjugated with muramyl dipeptide, adjuvant peptide.
  • the carrier sequence was as follows:
  • the initial starting scale of the synthesis was 0.01 mmole at level zero which then became a 0.16 mmole scale synthesis during the level four coupling cycles due to the doubling which occurred at each branch point.
  • Four Lys(boc) residues were inserted in intermediate level (2) as attachment sites for adjuvant peptide.
  • the fmoc protecting groups were initially left on the completed peptide in order to block the 16 terminal lysines' (32) -NH 2 groups from reacting with the adjuvant peptide during its attachment to the internal or intermediate lysines on level (2) .
  • the peptide was cleaved from the resin, and the boc protecting groups on the internal lysines were simultaneously removed using 40 ml of 25% TFA in DCM which contained 1.0 ml of water.
  • the spent resin was washed with 25% TFA in DCM and then with DCM.
  • the solutions were pooled and rotary evaporated three times.
  • the residual TFA was neutralized with diisopropylethylamine (DIEA) , and then the branched chain peptide was reacted with 0.16 mmole of MDP which is a four fold molar excess over the free internal lysine attachment sites on level (2) .
  • DIEA diisopropylethylamine
  • the (32) fmoc groups on the terminal (16) lysines were removed with 20% pyrrolidine in DCM. The solvents were removed by rotary evaporation three times. The material was diluted with DCM, neutralized with HOAc, and extracted twice with water to separate the peptide from the residual organics.
  • Initial purification of the muramyl dipeptide 4 /branched chain synthetic carrier peptide conjugate was achieved by gel filtration on a 180 ml column of Sephadex G-10 which was washed and equilibrated with 0.1 M ammonium bicarbonate pH 8.0. The fractions were analyzed.
  • EXAMPLE 4 Two totally synthetic immunogens were built using the PC-1580 of Example 1 and MDP 4 /Carrier of Example 3, and using PC-1581 of Example 2 and MDP 4 /Carrier of Example 3. Each molecule of MDP 4 /Carrier had a total of (32) free NH 2 s as the termini of the (16) branches of the carrier peptide. A three fold molar excess of HOBt/DIPCDI activated synthetic antigenic peptide, either PC-1580 or PC-1581 was used. 10 mg of PC-1580 was activated with a molar equivalent of HOBt and DIPCDI.
  • the activated antigenic peptide was then reacted with 0.22 mg (23.0 nmole) of MDP 4 /Carrier overnight in DMF with constant mixing. The reactions was stopped by the addition of water which hydrolyzed the activated esters, thus allowing for the excess antigenic peptide to be recovered for reutilization.
  • the PC-1580 32 /MDP 4 /Carrier synthetic immunogen (calculated Mol. Wt. 154,532) was isolated from the free antigenic peptide, PC-1580 (Mol. Wt. 4531) and low molecular weight organic molecules by gel filtration on Sephadex G-25 in 0.1 M acetic acid. The synthetic immunogen eluted in the void volume of the column.
  • the peptides were activated with EDAC and then conjugated to the protein in a two step reaction sequence.
  • Each reaction was performed using standard procedures and routine reaction conditions.
  • the conjugates were isolated from the reaction mixture by gel filtration on Sephadex G-25, dialyzed, and lyophilized. The yields based upon the weights of the final products were greater than 95% for both the PC-1580/thyroglobulin and PC-1581/thyroglobulin conjugates. Both of the peptide/protein conjugates were then used in conjunction with Freund's complete adjuvant to immunize animals.
  • ELISA was used to assess the response each of the immunogens had in eliciting the production of antibodies in each animals of all the test groups.
  • the ELISA conditions used for these assays are as follows.
  • the assays were set up on 96 well high binding microtiter plates as follows:
  • Row A Serial dilutions of pre-immune serum from one animal
  • Row B - D Serial dilutions of the antisera obtained from one animal of the group
  • Row E Serial dilutions of the pre-immune serum from another animal
  • Row F - H Serial dilutions of the antisera obtained from another animal of the group
  • One antiserum from each of the four groups of rabbits of Example 6 was selected for use in western immunoblot and IFA experiments.
  • Whole cell lysates from cells which had been transfected with the HIV-1 genome and which have been shown to produce various HIV-1 proteins were subjected to SDS-PAGE following standard procedures. After electrophoresis, the proteins were electrophoritically transferred to PVDF membranes and the membranes were blocked with non-fat milk. Alternating lanes on the membrane had either diluted primary antibody or diluted primary antibody that had been blocked by preincubation for 60 min. with 100 gm/ml of antigenic peptide, either PC-1580 or PC-1581. Both the primary and blocked primary antibody were allowed to bind for (2) hours.
  • the membrane strips were 'washed twice before affinity, purified HRP-goat anti-rabbit IgG second antibody was then bound for 60 minutes.
  • the membranes were washed four times.
  • the DAP/H 2 0 2 color reaction was performed.
  • Each of the four antisera bound specifically to two bands on the membranes. These proteins had molecular weights of 120 kD and 160 kD, respectively, and were identified as HIV-1 gp.120 and gpl60. No bands were detected on the strips to which the peptide blocked antibodies were applied.
  • EXAMPLE 8 The antibodies of Example 6 were used in indirect immunofluorescent assay (IFA) staining experiments on fixed whole cells which produce HIV-1 gpl20 and gpl60 in culture. In these IFA experiments the antibodies have been found to bind specifically to HIV-1 transfected cells. They do not, however, bind to non-transfected normal cells, and the binding can be blocked by pre-incubating the diluted primary antibody with antigen peptide before it is applied to the fixed cells. The proteins in this Example had not been denatured as they were in the western immunoblots following SDS-PAGE, in Example 7.
  • IFA indirect immunofluorescent assay

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Abstract

L'invention concerne un vecteur synthétique pour fournir au moins un composant biologiquement actif à un organisme. Ce vecteur est constitué par un peptide synthétique qui peut être lié à une résine. Le peptide vecteur porte un acide aminé terminal ayant une paire de sites fonctionnels terminaux qui sont capables de fixer au moins une paire de composants et en particulier de composants biologiquement actifs. De plus, des acides aminés additionnels peuvent être fixés à l'extrémité de la chaîne ramifiée d'acides aminés pour former une matrice ayant un nombre progressivement croissant de ramifications et de sites de fixation. Des composés biologiquement actifs peuvent se fixer aux ramifications multiples de la chaîne avec un haut degré de précision. De plus, des sites de fixation intermédiaires additionnels de la matrice peuvent être utilisés pour fixer d'autres groupes fonctionnels tels que des adjuvants peptidiques. La chaîne synthétique ramifiée servant de vecteur peut être employée pour réaliser un immunogène synthétique de très haute pureté, avec des rapports spécifiques de couplage pour les différents composés fixés aux extrémités de chaînes ramifiées et à d'autres sites pré-définis d'attachement intermédiaire. Ainsi, le vecteur synthétique de la présente invention permet de fournir des combinaisons spécifiques de composants biologiquement actifs.
PCT/US1994/005981 1993-06-18 1994-05-26 Vecteur et immunogene de synthese WO1995000540A1 (fr)

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US08/080,186 1993-06-18

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Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0670728A1 (fr) * 1992-11-12 1995-09-13 Molecular Dynamics, Inc. Porteurs lipophiles a base de peptides destines a l'administration ciblee de medicaments selon un concept rationnel de fixation des medicaments
EP0884327A1 (fr) * 1997-06-11 1998-12-16 The School Of Pharmacy, University Of London Polypeptides dendritiques à base de lysine pour apport ciblé de médicaments
WO2000075171A1 (fr) * 1999-06-04 2000-12-14 Avecia Limited Procede de preparation de supports pour synthese en phase solide
WO2001007469A3 (fr) * 1999-07-23 2001-05-10 Antonio Verdini Dendrimeres polypeptidiques en tant que porteurs unimoleculaires d'agents de contraste d'imagerie diagnostique, de substances bioactives et de medicaments
US6774102B1 (en) * 1999-09-29 2004-08-10 Gambro Dialysatoren Gmbh & Co. Kg Extracorporeal endotoxin removal method

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EP0339695A2 (fr) * 1988-04-29 1989-11-02 Stichting Voor De Technische Wetenschappen Procédé de préparation d'un conjugué antigénique ou immunogénique ainsi que l'utilisation de tels conjugués
EP0343460A2 (fr) * 1988-05-24 1989-11-29 F. HOFFMANN-LA ROCHE & CO. Aktiengesellschaft Peptides cs de P. Falciparum comme épitope universel de cellules t
GB2228262A (en) * 1989-01-25 1990-08-22 Nat Inst Immunology Antigenic derivative of GnRH

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4289872A (en) * 1979-04-06 1981-09-15 Allied Corporation Macromolecular highly branched homogeneous compound based on lysine units
EP0339695A2 (fr) * 1988-04-29 1989-11-02 Stichting Voor De Technische Wetenschappen Procédé de préparation d'un conjugué antigénique ou immunogénique ainsi que l'utilisation de tels conjugués
EP0343460A2 (fr) * 1988-05-24 1989-11-29 F. HOFFMANN-LA ROCHE & CO. Aktiengesellschaft Peptides cs de P. Falciparum comme épitope universel de cellules t
GB2228262A (en) * 1989-01-25 1990-08-22 Nat Inst Immunology Antigenic derivative of GnRH

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Title
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Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0670728A1 (fr) * 1992-11-12 1995-09-13 Molecular Dynamics, Inc. Porteurs lipophiles a base de peptides destines a l'administration ciblee de medicaments selon un concept rationnel de fixation des medicaments
EP0670728A4 (fr) * 1992-11-12 1996-04-17 Molecular Dynamics Inc Porteurs lipophiles a base de peptides destines a l'administration ciblee de medicaments selon un concept rationnel de fixation des medicaments.
EP0884327A1 (fr) * 1997-06-11 1998-12-16 The School Of Pharmacy, University Of London Polypeptides dendritiques à base de lysine pour apport ciblé de médicaments
US6194543B1 (en) 1997-06-11 2001-02-27 The School Of Pharmacy University Dendritic polypeptides
WO2000075171A1 (fr) * 1999-06-04 2000-12-14 Avecia Limited Procede de preparation de supports pour synthese en phase solide
US6750312B1 (en) 1999-06-04 2004-06-15 Avecia Limited Process for the preparation of supports for solid phase synthesis
WO2001007469A3 (fr) * 1999-07-23 2001-05-10 Antonio Verdini Dendrimeres polypeptidiques en tant que porteurs unimoleculaires d'agents de contraste d'imagerie diagnostique, de substances bioactives et de medicaments
AU772167B2 (en) * 1999-07-23 2004-04-08 Les Laboratoires Servier Polypeptide dendrimers as unimolecular carriers of diagnostic imaging contrast agents, bioactive substances and drugs
US6774102B1 (en) * 1999-09-29 2004-08-10 Gambro Dialysatoren Gmbh & Co. Kg Extracorporeal endotoxin removal method

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