WO2000035427A1 - Techniques de reduction de l'antigenicite de proteines, de tissus ou d'organes - Google Patents

Techniques de reduction de l'antigenicite de proteines, de tissus ou d'organes Download PDF

Info

Publication number
WO2000035427A1
WO2000035427A1 PCT/US1999/029452 US9929452W WO0035427A1 WO 2000035427 A1 WO2000035427 A1 WO 2000035427A1 US 9929452 W US9929452 W US 9929452W WO 0035427 A1 WO0035427 A1 WO 0035427A1
Authority
WO
WIPO (PCT)
Prior art keywords
protein
collagen
tissue
acylating agent
immunogenicity
Prior art date
Application number
PCT/US1999/029452
Other languages
English (en)
Inventor
Dale P. Devore
Original Assignee
Collagenesis, Inc.
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 Collagenesis, Inc. filed Critical Collagenesis, Inc.
Publication of WO2000035427A1 publication Critical patent/WO2000035427A1/fr

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/78—Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG]
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
    • C07K1/107—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length by chemical modification of precursor peptides
    • C07K1/1072—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length by chemical modification of precursor peptides by covalent attachment of residues or functional groups
    • C07K1/1077—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length by chemical modification of precursor peptides by covalent attachment of residues or functional groups by covalent attachment of residues other than amino acids or peptide residues, e.g. sugars, polyols, fatty acids

Definitions

  • this invention relates to methods for reducing the immunogenicity of proteins, tissues, processed tissues, tissue constructs, or whole organs, for example, as a means to facilitate their implantation into an animal recipient.
  • Surgical implants find a number of uses in the medical field, including burn treatment, skin grafting, dermal contouring, soft tissue reconstruction, and bone, ligament, and tendon repair.
  • an implant is composed of a material that is immunologically inert and retains its implant properties for the life of the patient.
  • Biocompatible materials such as silicone or collagen are available.
  • collagen for example, in various forms including commercially available forms designed for surgical implantation, has been found to be somewhat immunogenic.
  • implant materials such as collagen
  • some implant procedures involve multiple treatments to either correct a defect or maintain a desired result, and certain patients, after multiple exposures to an implant, go on to develop immune responses (DeLustro et al, Plast. Reconstr. Surg., 79(4): 581- 594 (1987)).
  • patients are sometimes exposed to more than one implant material, and there exists evidence that implant materials can sometimes act in synergy to provoke an immune response.
  • collagen or silicone implants alone provoked only a minimal immune response; however, when co-introduced into an animal, these materials were notably immunogenic (Takayama et al., J. Laryngol.
  • Alport's syndrome Another debilitating variation of Goodpasture's syndrome, termed Alport's syndrome, develops in 15%) of patients (Fleming et al, Transplantation, 46(6):857-859 (1988)). Patients with this X-linked recessive disease produce collagen autoantibodies against an immunogenic epitope of the alpha 5 chain of type IN collagen and suffer from sensorineural deafness, nephritis, and ocular malformations (Weber et al., Clin. Investig., 70(9):809-815 (1992)).
  • the present invention provides a method for generating a substantially non-immunogenic protein suitable for implanting into an animal recipient.
  • This method involves acylating the protein by exposing the protein to an acylating agent under conditions that preferably allow for at least one free amine group (and more preferably, 90% of the amine groups) to be acylated.
  • the acylating agents derivatize a deprotonated free amine group of the protein with an anionic group to reduce immunogenicity.
  • This derivatizing step preferably reduces the pKa of the protein.
  • Any acylating agent may be employed to carry out the methods of the invention, but preferred acylating agents include anhydrides (for example, succinic anhydride), acid chlorides, and sulfonyl chlorides.
  • the protein rendered non- immunogenic by acylation is collagen (for example, type I or type II collagen) or a protein associated with a tissue, processed tissue, tissue construct, or whole organ (for example, skin, cartilage, ligaments, tendons, muscle, nervous tissue, islet cells, corneal tissue, heart valves, blood vessels, bone marrow, peripheral blood, heart, lung, liver, kidney, adrenal gland, pancreas, or intestine).
  • the acylated protein, tissue, processed tissue, tissue construct, or whole organ is suitable for implanting into a mammal, and, more preferably, into a human.
  • the mammal preferably exhibits no substantial production of antibodies to the acylated protein.
  • the invention also provides a method for reducing an immunogenic response to an implanted collagen, preferably, a heterologous collagen, by exposing the collagen to an acylating agent which causes acylation of the collagen to occur. This method is useful for reducing an immunogenic response such as arthritis.
  • the invention provides a method for generating a non-immunogenic protein, for example, collagen that is derived from a primate, cow, pig, sheep, or, preferably, a human.
  • a non-immunogenic protein for example, collagen that is derived from a primate, cow, pig, sheep, or, preferably, a human.
  • protein is meant a compound composed of two or more linked amino acids, synthesized in vitro or in vivo, and existing as a monomer or multimer.
  • proteins of the extracellular matrix for example, collagen types I-XIX or a combination thereof, elastin, fibronectin, and proteoglycans
  • structural proteins for example, cytoskeletal proteins
  • enzymatic proteins for example, cell surface proteins (for example, MHC class I proteins, MHC class II proteins, LFA proteins, integrins, and ICAM proteins), and ligands (for example, peptide hormones and cytokines).
  • immunogenic is meant the capacity to stimulate an immune response (for example, a humoral response or cell-mediated response) in an animal.
  • acylated protein is meant a protein in which at least one free amine group has been modified by exposure to an acylating agent.
  • acylating agent is meant an agent that transfers an acyl group to another nucleophile.
  • acylating agents include anhydrides, acid chlorides, and sulfonyl chlorides.
  • heterologous is meant a protein that is foreign in origin to the recipient animal.
  • collagen is meant any of the collagens (for example, type I-XIX or a mixture, variant, or fragment thereof) derived from a multicellular organism (see, e.g., Prockop et al., Annu. Rev. Biochem., 64:403-434 (1995)).
  • the term includes collagen in any form, whether produced naturally, recombinantly, or treated mechanically, chemically, or enzymatically.
  • biomaterial any material that includes a protein, cell, tissue, processed tissue, tissue construct, or organ.
  • a processed tissue may include addition manipulation beyond the method of the invention (e.g., to facilitate, harvesting, implanting, tolerance, or surgical success).
  • a tissue construct may involve further manipulation of the tissue such that a particular structural feature is obtained (e.g., skin being grown or shaped for a particular graft application or endothelial cells being grown or shaped into tubes for use as blood vessels).
  • implant is meant any biomaterial which is introduced into an animal and which has been derived from a source other than that animal recipient.
  • one advantage of the present technique is that the immunogenicity of proteins may be reduced by acylating lysine and arginine residues. Since most proteins have lysine and arginine residues, the method of the invention is suitable for reducing the immunogenicity of virtually any protein, or any protein associated with a tissue or organ. In addition, this technique is relatively inexpensive and facilitates the reduction of immunogenicity with minimal processing of the protein, tissue, or organ. Other features and advantages of the invention will be apparent from the following detailed description and from the claims.
  • immunogenicity is reduced by acylation with succinic anhydride.
  • any acylating agent may be used for this purpose, and, in particular examples, any anhydride, acid chloride, or sulfonyl chloride may be utilized. All of these acylating agents work by reacting with, and derivatizing, the deprotonated free amines of a protein. In many cases, the free amine group is derivatized with a chemical moiety that provides an anionic group, thereby reducing the pKa of the protein. For certain proteins, such as collagen, this also renders the protein more soluble at physiological pH.
  • Amine-reactive acylating agents useful in the invention include, without limitation, carboxylic acid anhydrides (e.g., glutaric anhydride, benzoic anhydride, and 1,2,4,5-benzene tertracarboxylic acid dianhydride), carboxylic acid esters (e.g., monophenyl terephthalate, ethyl benzoate, and alpha- naphthoic acid ethyl ether), carboxylic acid halides (e.g., succinic acid chloride), sulfonic acids (e.g., 1,3,-benzene-disulfonic acid, aniline-2-sulfonic acid, and 3-nitrobenzene-sulfonic acid), 2-formylbenzene-sulfonic acid halides (e.g., 4,4' biphenyl-disulfonyl chloride and benzene sulfonyl chloride), and mixtures thereof.
  • carboxylic acid anhydrides
  • the acylating agent may be an aliphatic or aromatic, mono-, di-, or higher functional carboxylic acid anhydride, ester or halide; or sulfonic acid or halide, such as a lower alkanoic, lower alkane-dioic, or higher functional lower alkane carboxylic, or aryl mono-, di-, or higher functional carboxylic (e.g., benzoic or naphthoic), acid anhydride, ester or halide, or lower alkyl, or aryl (e.g., phenyl or naphthyl), mono-, di-, or higher functional sulfonic acid or halide, to provide the corresponding acyl (carbonyl or sulfonyl) moiety on the amine group, for example, lower alkanoyl, aroyl (e.g., phenoyl or naphthoyl), alkyl sulfonyl, aryl (e
  • the acylating agent may be added directly to a reaction mixture as a solid material (e.g., a powder) or dissolved in a suitable organic solvent such as acetone, N,N-dimethylformamide (DMF), ethanol, or methyl pyrrolidone.
  • a suitable organic solvent such as acetone, N,N-dimethylformamide (DMF), ethanol, or methyl pyrrolidone.
  • the total quantity of acylating agent depends on the extent of modification required or desired. In the case of collagen, the quantity of acylating agent required should generally satisfy the weight ratio of acylating agent to wet tissue of approximately 0.005-0.5:1, and preferably 0.05-0.1 : 1.
  • the protein acylation reaction (i.e., the amine modifying reaction) generally proceeds within a pH range of 7 to 11, although it is preferably carried out at a mildly basic pH (for example, pH 8-10, and, more preferably 8.5-9) to increase the reaction speed and reduce the processing time.
  • a mildly basic pH for example, pH 8-10, and, more preferably 8.5-9
  • an acylation buffer at physiological pH is preferably utilized.
  • the acylation methods provided in any of DeVore et al. and Kelman et al. U.S. Pat. Nos. 5,412,135; 5,104,957; 5,201,764; and 5,480,427) may also be utilized to reduce protein immunogenicity, and these patents are incorporated herein by reference.
  • EXAMPLE 1 Methods for Reducing the Immunogenicity of a Protein
  • the present invention provides a technique for reducing the immunogenicity of an implant through protein acylation.
  • the acylation occurs via free amine groups found on the protein's lysine and arginine residues.
  • a type I bovine collagen sample was prepared and acylated using standard procedures (as similarly described in U.S. Pat. Nos. 5,104,957; 5,492,135; and 5,631,243). Briefly, collagen containing bovine dermis was diced with a scalpel and, in 5g aliquots in 20 ml volumes of deio ized water, throughly macerated with a dispersion tool. Each aliquot was poured into a 50 ml centrifuge tube, diluted with 10 ml of deionized water, shaken vigorously, and centrifuged at 4000 rpm for 10 minutes at 4°C.
  • the tubes were decanted, and the material was consolidated and resuspended in a 350 ml volume of 0.5M NaOH in a 500 ml container and rotated for one hour at 4°C.
  • the sample was then divided into 50 ml tubes, centrifuged as above, rinsed twice with 50 mM sodium phosphate buffer, and combined in a 500 ml container with 350 ml of a 50 mM sodium phosphate buffer.
  • This solution was titrated with 0.5M phosphoric acid to pH 2.5, rocked for three hours at 4°C, and then titrated with 0.5M NaOH to pH 7.1.
  • the sample was again divided, centrifuged as above, and washed three times with a 4 mM phosphate buffer.
  • the resultant material was homogenized in 5.0 g aliquots in 20 ml of 20 mM disodium phosphate buffer (pH 9.0) using the "beaker in a beaker” method to disperse the bovine collagen matrix. This material was then derivatized with three additions (1.85 ml) of 0.08% glutaric anhydride solution (the pH being adjusted to 9.0 between additions). The material was centrifuged and treated again with a glutaric anhydride solution ( 1 ml) and retrieved.
  • the material was then diluted with 4 mM phosphate buffer (pH 7J), filtered (100 ⁇ m pore size), centrifuged (at 10,000 rpm for 10 minutes in SS-34 rotor), and adjusted to a concentration of about 30 mg/ml.
  • immunologically naive rats were immunized with a collagen sample (at approximately 30 mg/ml in Freund's adjuvant) derivatized with glutaric anhydride as described in Example 1.
  • An untreated collagen control i.e., Zyderm ITM, a non-derivatized, commercially available collagen
  • Zyderm ITM Zyderm ITM, a non-derivatized, commercially available collagen
  • lymphocytes were purified from the lymph nodes. These lymphocytes were then tested in vitro for their ability to proliferate when incubated with the same collagen sample used to immunize the test animal.
  • Lymphocyte proliferation was assayed by measuring the amount of 3 H- thymidine incorporated by the cells after co-incubation with either treated or untreated collagen.
  • Parallel experiments were performed with adjuvant (complete Freund's adjuvant) in order to set a baseline against which the immunogenic potential of the various collagen products were measured.
  • test animals may be immunized with treated and untreated collagen to determine if the implants cause induce antibody formation. Further, using established methods, such animals may also be monitored for the occurrence of any delayed type hypersensitivity response (DTH) at the immunization site.
  • DTH delayed type hypersensitivity response
  • peripheral venous blood may be drawn from human subjects and tested in vitro for the presence of any T cells responsive to an implant.
  • studies are performed either in parallel or in addition to other appropriate tests (e.g., skin testing).
  • heparinized venous blood is obtained from the patient, and peripheral blood mononuclear cells are separated by density gradient centrifugation (for example, by Ficoll-HypaqueTM density gradient centrifugation), washed twice in RPMI media, counted, and resuspended in an appropriate cell culture medium (RPMI 1640 (Biowhittaker, Walkersville, MD), 10% normal human serum (Normalceraplus, NABI, Miami, FL), lOmM Hepes buffer, 100 ⁇ g/ml penicillin, and streptomycin)).
  • density gradient centrifugation for example, by Ficoll-HypaqueTM density gradient centrifugation
  • RPMI 1640 Biowhittaker, Walkersville, MD
  • 10% normal human serum Normalceraplus, NABI, Miami, FL
  • lOmM Hepes buffer 100 ⁇ g/ml penicillin, and streptomycin
  • Lymphocyte proliferation is assayed by measuring the amount of 3 H-thymidine incorporated by the cells (added for the final 18 hours of culture) using a ⁇ scintillation counter (Beta Plate, Wallac, Gaithersburg, MD) as previously described in Vella et al. (Transplantation, 64 (6): 795-800 (1997)).
  • blood samples from a patient are drawn prior to the introduction of an implant and at various time points thereafter so the clinician can assess how well the patient tolerates the implant over time.
  • this assay can assist the clinician in determining which implants are tolerated and which are immunogenic.
  • Type I collagen samples were prepared from bovine hide using the following procedure as similarly described in U.S. Pat. Nos. 5,354,336, 5,631,243, and 5,476,515. Briefly, bovine hide was cut into pieces approximately l x l cm and washed in 0.2M NaCl for 1 hour with stirring. These pieces were then rinsed in 20 volumes of deionized water and subsequently added to 10 volumes of 0.5M acetic acid at 4°C. After 72 hours, the resultant viscous mass was filtered through cheesecloth to remove any insoluble material.
  • the filtrate was then diluted with an equal volume of 0.5M acetic acid (1 : 1) and digested with pepsin (Sigma Chemical Co.; 1 :60,000), added to a concentration of 2% (w/w), for 48 hours at 4°C. Any undigested hide was removed by centrifugation at 20,000 x g, and the remaining digested collagen was precipitated using dialysis against 0.01M disodium phosphate. Next, the collagen precipitate was redissolved in 0.5M acetic acid and then reprecipitated by the addition of NaCl to 0.8M. The resultant pellet was redissolved in 0.5M acetic acid, and the pH was adjusted to 9.5 by the careful addition of ION and IN NaOH at 2-4°C.
  • pepsin Sigma Chemical Co.
  • a treated collagen sample was made by derivatizing the above sample by acylation using succinic anhydride (Aldrich Chemical Co.).
  • 500 mis of soluble collagen at a concentration of about 3 mg/ml was adjusted to pH 9.0 using ION and IN NaOH.
  • solid succinic anhydride was added to a concentration of 10% (w/w) collagen solids.
  • This mixture was allowed to react at pH 9.0 for 30 minutes, and the reaction was stopped by raising the pH to 12.0 using IN NaOH.
  • the solution was filtered through a 0.45 ⁇ filter, and the pH was adjusted to 4.3 using 6N and IN HCl.
  • the mixture was stirred for 10 minutes as the derivatized collagen precipitated.
  • the treated collagen precipitate was then dissolved in 0.05M sodium phosphate buffer at pH 12 to a concentration of 5 mg/ml. This treated collagen solution was stored at 2-8°C and designated as lot 108.
  • the multi-well plates were then incubated with a 10% BSA solution for 1 hour at room temperature, decanted, and washed three times or more with PBS (until visible protein bubbles disappeared).
  • 25 ⁇ l of antisera, from either a test animal or control animal was added to each well and allowed to incubate for 1 hour at room temperature.
  • Wells were then washed three times with PBS and exposed to 25-50 ⁇ l of 12 I-labeled protein A (at least 50,000 CPM/well).
  • Wells were then washed three times with PBS to remove non-specific counts.
  • Samples were harvested with 1 N NaOH and assayed for the presence of anti-collagen antibodies in a gamma counter. Unless otherwise described, the numbers presented indicate counts per minute (CPM) emitted by 125 I-labeled protein A, revealing the amount of anti-collagen antibodies bound to a particular collagen substrate (Tables 2 and 3).
  • Tables 2 and 3 represent the results of these radioimmunoassays. Each table indicates the presence of antibodies capable of binding a particular collagen substrate. The substrates tested in Tables 2 and 3 are treated (lot 108) and untreated type I bovine collagen.
  • anti-collagen antibody levels in mice immunized with acylated bovine collagen were similar to levels measured in mice immunized with only a control buffer.
  • the antibody titers measured for animals immunized with treated collagen were 90% of the antibody titers measured for the acetic acid negative control.
  • acylated collagen was significantly less antigenic than untreated collagen is shown in Table 2, where acylated bovine collagen (lot 108) showed a 2-6 fold lower cross reactivity with various other anti- collagen antibodies than did untreated material (compare, for example, shaded boxes of Tables 2 and 3).
  • acylated bovine collagen (lot 108) showed a 2-6 fold lower cross reactivity with various other anti- collagen antibodies than did untreated material (compare, for example, shaded boxes of Tables 2 and 3).
  • antibody titers measured for animals immunized with treated collagen were only 46% of the antibody titers measured for animals immunized with untreated collagen, representing a 54% reduction in the antibody titers for the derivatized collagen.
  • untreated collagen induced antibody titers that were 196% higher than the acetic acid controls.
  • biomaterials are obtained and prepared by standard medical techniques. These biomaterials are then treated by the acylation techniques described above.
  • acylation conditions designed to reduce or prevent cell lysis are preferably employed, including acylation reactions carried out at physiological pH.
  • Biomaterials that have been acylated may be tested for reduced immunogenicity using any standard in vitro immunoassay technique.
  • a reduction in the immunogenicity of a biomaterial may also be tested in an animal model, with appropriate modifications, for example, as described herein.
  • the biomaterial can be introduced into the recipient animal or patient using appropriate surgical techniques.
  • Unused non-immunogenic biomaterial may be tissue-banked for future applications.
  • One particular biomaterial amenable to this method is skin which can be grown in large sheets in vitro using donor cells. This skin tissue is then removed from culture and processed using the methods of the invention. Essentially, the sheets of skin are dipped in a compatible buffer (for example, at physiological pH) in which an acylation reaction can proceed without compromising the integrity of the tissue.
  • Cartilage, ligaments, tendons, corneal tissue, heart valves, and blood vessels may also be harvested and acylated in this manner.
  • bone marrow or peripheral blood may be treated using a similar method.
  • Bone marrow or peripheral blood may be harvested according to conventional methods, centrifuged, and resuspended in a buffer that maintains the integrity of the blood cells while allowing acylation of immunogenic cell surface proteins.
  • the reaction is allowed to proceed for a period of time appropriate for reducing immunogenicity.
  • the reaction is then stopped, the cells resuspended in an appropriate physiological buffer, and the resuspended cells introduced into an animal recipient.
  • This method is particularly advantageous for treating peripheral blood for use in transfusions when blood typing is unavailable or not practical.
  • the blood group antigens may be rendered non-immunogenic using the acylation reaction of the invention. Accordingly, the resultant blood product can then be administered to any patient without concern for the patient's blood type.
  • Organ transplantation may also be facilitated by the methods of the invention.
  • Techniques for manipulating organs ex vivo for a limited time (24- 48 hours), for example, using hypothermic conditions and maintenance buffers, have been established. These conditions can be adapted to include an acylation step to reduce the immunogenicity of the organ.
  • Organs which may be treated by this technique include heart, lung, liver, kidney, adrenal gland, pancreas, and intestine.
  • the organ is perfused with a preservation solution designed to maintain high oncotic pressure and the maintenance of a near-intracellular ionic concentration.
  • Perfusion conditions can be modified to bathe the organ with an acylating reagent.
  • Acylation conditions may be modified depending on the degree of donor to recipient mismatch, with a higher degree of mismatch warranting more extended acylation.
  • the organ is reperfused with preservation solutions to ready the organ for implanting into a patient. The remaining steps of the transplant operation are carried out according to conventional transplant protocols.
  • This general method represents an improvement to conventional transplant procedures.
  • the method reduces graft rejection mediated by humoral or cellular immune responses.
  • implanting a tissue or organ of reduced immunogenicity requires the use of fewer immunosuppressive drugs to insure graft survival.
  • These drugs while essential for conventional transplantation success involving the use of allografts or xenografts, carry the dangerous side effects of liver toxicity and the risk of opportunistic infections. These side effects can be lessened or eliminated using the methods of the invention.
  • the methods described herein are used for the treatment of human patients, but may also be used to treat any other mammal, for example, any pet or livestock.
  • the methods of the invention may also be used to reduce the immunogenicity of any immunogenic protein or compound having free amine groups capable of being acylated, for example, any implant composed of, or coated with, a compound or protein having free amine groups.
  • this method may be used to reduce the immunogenicity of other immunogenic proteins including, without limitation, MHC class I proteins, MHC class II proteins, integrins, LFA proteins, and ICAM proteins.
  • acylation reactions would be carried out on these proteins as described herein.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Molecular Biology (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Medicinal Chemistry (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Zoology (AREA)
  • Toxicology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Peptides Or Proteins (AREA)

Abstract

L'invention concerne une technique qui permet de produire une protéine sensiblement non immunogène convenant à l'implantation dans un animal. La technique consiste à exposer la protéine à un agent d'acylation dans des conditions qui permettent d'obtenir l'acylation de la protéine.
PCT/US1999/029452 1998-12-15 1999-12-13 Techniques de reduction de l'antigenicite de proteines, de tissus ou d'organes WO2000035427A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US21160898A 1998-12-15 1998-12-15
US09/211,608 1998-12-15

Publications (1)

Publication Number Publication Date
WO2000035427A1 true WO2000035427A1 (fr) 2000-06-22

Family

ID=22787635

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1999/029452 WO2000035427A1 (fr) 1998-12-15 1999-12-13 Techniques de reduction de l'antigenicite de proteines, de tissus ou d'organes

Country Status (1)

Country Link
WO (1) WO2000035427A1 (fr)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20120109293A1 (en) * 2005-12-20 2012-05-03 Grandhope Biotech Co., Ltd. Biological artificial cornea and method of making
WO2012070971A1 (fr) * 2010-11-22 2012-05-31 Farber Boris Slavinovich Composition cosmétologique et pharmaceutique pour le rajeunissement et la régénération de la peau, y compris après les opérations chirurgicales
US11259959B1 (en) 2020-11-03 2022-03-01 D&D Biopharmaceuticals, Inc. Devices and methods for cornea treatment
US11938092B1 (en) 2022-11-30 2024-03-26 D&D Biopharmaceuticals, Inc. Devices and methods for cornea treatment

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4713446A (en) * 1985-09-06 1987-12-15 Minnesota Mining And Manufacturing Company Viscoelastic collagen solution for ophthalmic use and method of preparation
US4851513A (en) * 1985-09-06 1989-07-25 Minnesota Mining And Manufacturing Company Viscoelastic collagen solution for opthalmic use and method of preparation
US4883864A (en) * 1985-09-06 1989-11-28 Minnesota Mining And Manufacturing Company Modified collagen compound and method of preparation

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4713446A (en) * 1985-09-06 1987-12-15 Minnesota Mining And Manufacturing Company Viscoelastic collagen solution for ophthalmic use and method of preparation
US4851513A (en) * 1985-09-06 1989-07-25 Minnesota Mining And Manufacturing Company Viscoelastic collagen solution for opthalmic use and method of preparation
US4883864A (en) * 1985-09-06 1989-11-28 Minnesota Mining And Manufacturing Company Modified collagen compound and method of preparation

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
HABEEB ET AL.: "Molecular Structural Effects Produced in Proteins by Reaction with Succinil Anhydride", BIOCHIM. BIOPHYS. ACTA, vol. 29, 1958, pages 587 - 593, XP002924778 *
PUGH ET AL.: "Alteration of Protective and Serologic Responses in BALB/c Mice Vaccinated with Chemically Modified versus Nonmodified Proteins of Brucella Abortus 19", INFECTION AND IMMUNITY, vol. 62, no. 12, 1994, pages 5327 - 5334, XP002924779 *

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20120109293A1 (en) * 2005-12-20 2012-05-03 Grandhope Biotech Co., Ltd. Biological artificial cornea and method of making
WO2012070971A1 (fr) * 2010-11-22 2012-05-31 Farber Boris Slavinovich Composition cosmétologique et pharmaceutique pour le rajeunissement et la régénération de la peau, y compris après les opérations chirurgicales
US11259959B1 (en) 2020-11-03 2022-03-01 D&D Biopharmaceuticals, Inc. Devices and methods for cornea treatment
US11938092B1 (en) 2022-11-30 2024-03-26 D&D Biopharmaceuticals, Inc. Devices and methods for cornea treatment
US12102598B2 (en) 2022-11-30 2024-10-01 D&D Biopharmaceuticals, Inc. Devices and methods for cornea treatment

Similar Documents

Publication Publication Date Title
US5002071A (en) Injectable soft tissue augmentation materials from the placenta and their method of manufacture
CA1339092C (fr) Methode pour le traitement des greffes avant la transplantation
RU2104703C1 (ru) Способ получения материала для остеопластики и полученный этим способом материал
US4280954A (en) Crosslinked collagen-mucopolysaccharide composite materials
EP0419111B1 (fr) Compositions à base de collagène et méthode pour leur préparation
EP1255577B1 (fr) Matrice cellulaire
CA1087610A (fr) Substances composees de mucopolysaccharides et de collagene reticules
US4776853A (en) Process for preparing biological mammalian implants
US6743435B2 (en) Processing animal tissues by decellularizing, increasing surface area and acylating
JP2834155B2 (ja) コラーゲンフレーク体
JPH04502336A (ja) 骨形成具
JP2002537896A (ja) 可移植性軟骨組織のｉｎｖｉｔｒｏでの製造
CN85101396A (zh) 可被生物降介的衬质和生产它的方法
FR2889449A1 (fr) Preparations implantables
JPS58121958A (ja) 軟組織強化用コラ−ゲン移植物質及び移植方法
JP2004267791A5 (fr)
JPH03504736A (ja) 移植用骨コラーゲンマトリックス
JPH11503338A (ja) 軟骨組織再構成のための再吸収可能な細胞外マトリックス
JPH11501514A (ja) 新規の封入組成物及び封入方法
US5138030A (en) Process for extracting type I collagen form an avian source, and applications therefor
US5916553A (en) Complex for inducing bone growth in the mastoid cavity
JPH10501706A (ja) 細胞−ゲル
WO2000035427A1 (fr) Techniques de reduction de l'antigenicite de proteines, de tissus ou d'organes
JP3008034B2 (ja) 体内注入可能な高濃度架橋化アテロコラ−ゲン移植用組成物
Villa et al. Residual heteroantigenicity of glutaraldehyde‐treated porcine cardiac valves

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): BR CA JP

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE

121 Ep: the epo has been informed by wipo that ep was designated in this application
DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
122 Ep: pct application non-entry in european phase