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WO2007010552A2 - Conjugue d'erythropoietine peg n-terminal - Google Patents

Conjugue d'erythropoietine peg n-terminal Download PDF

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Publication number
WO2007010552A2
WO2007010552A2 PCT/IN2006/000103 IN2006000103W WO2007010552A2 WO 2007010552 A2 WO2007010552 A2 WO 2007010552A2 IN 2006000103 W IN2006000103 W IN 2006000103W WO 2007010552 A2 WO2007010552 A2 WO 2007010552A2
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WO
WIPO (PCT)
Prior art keywords
epo
conjugate
erythropoietin
peg
mpeg
Prior art date
Legal status (The legal status 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 status listed.)
Ceased
Application number
PCT/IN2006/000103
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English (en)
Other versions
WO2007010552B1 (fr
WO2007010552A3 (fr
Inventor
Subhash V. Kapre
Umesh S. Shaligram
Kwang Nho
Gunwon Bae
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Serum Institute of India Pvt Ltd
Sunbio Inc
Original Assignee
Serum Institute of India Pvt Ltd
Sunbio Inc
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Filing date
Publication date
Application filed by Serum Institute of India Pvt Ltd, Sunbio Inc filed Critical Serum Institute of India Pvt Ltd
Publication of WO2007010552A2 publication Critical patent/WO2007010552A2/fr
Publication of WO2007010552A3 publication Critical patent/WO2007010552A3/fr
Publication of WO2007010552B1 publication Critical patent/WO2007010552B1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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Classifications

    • 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
    • A61K47/56—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 an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule
    • A61K47/59—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 an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes
    • A61K47/60—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 an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P13/00—Drugs for disorders of the urinary system
    • A61P13/12—Drugs for disorders of the urinary system of the kidneys
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • the present invention relates to a conjugated Erythropoietin and a method for producing thereof.
  • Erythropoiesis is the production of red blood cells, which occurs to offset cell destruction. Erythropoiesis is a controlled physiological mechanism that enables sufficient red blood cells to be available for proper tissue oxygenation.
  • Naturally occurring human erythropoietin (hEPO) is produced in kidney and is the humoral plasma factor which stimulates red blood cell production (Carnot, P and Deflandre, C (1906) CR. Acad. Sci. 143: 432; Erslev, A J (1953 Blood 8: 349; Reissmann, K R (1950) Blood 5: 372; Jacobson, L O, Goldwasser, E, Freid, W and Plzak, L F (1957) Nature 179: 6331-4).
  • Naturally occurring EPO stimulates the division and differentiation of committed erythroid progenitors in the bone marrow and exerts its biological activity by binding to receptors on erythroid precursors (Krantz, B S (1991) Blood 77: 419).
  • Erythropoietin has been manufactured biosynthetically using recombinant DNA technology (Egrie, J C, Strickland, T W, Lane, J et al. (1986) Immunobiol. 72: 213-224) and is the product of a cloned human EPO gene inserted into and expressed in the ovarian tissue cells of the Chinese hamster (CHO cells).
  • the carbohydrate groups account for approximately 40% of the molecular weight that glycosylate the protein at glycosylation sites on the protein (Sasaki, H, Bothner, B, Dell, A and Fukuda, M (1987) J. Biol. Chem. 262: 12059).
  • EPO is used in the treatment of anemia in chronic renal failure patients (CRF) (Eschbach, J W, Egri, J C, Downing, M R et al. (1987) NEJM
  • PEG polyethylene glycol compound
  • European Patent Application EP 651,761 discloses the selective modification of recombinantly produced polypeptides at terminal alpha- carbon reactive groups.
  • the first step in the method is to form a recombinantly produced polypeptide so that it is protected at the terminal alpha-carbon reactive group with a biologically added protecting group.
  • the biologically added protecting group is preferably an amino acid, peptide and/or polypeptide that contains at least one site that is cleavable enzymatically or chemically, and preferably has a sequence that is not present in the sequence of the desired polypeptide.
  • the biologically protected polypeptide is reacted with chemical protecting agents to protect the side chain groups and then is cleaved with a cleavage reagent specific for the biologically added protecting group.
  • chemical protecting agents to protect the side chain groups and then is cleaved with a cleavage reagent specific for the biologically added protecting group.
  • a polypeptide is produced having an unprotected N-terminal amino group and protected side chain reactive groups.
  • the unprotected N-terminal amino group is modified with a modifying agent to form an N-terminally modified and side-chain- protected polypeptide. It is then deprotected to form an N-terminally modified polypeptide.
  • EP 651 ,761 suggests that any sequence of amino acids may be attached as biologically added protecting groups.
  • EPO is expressed with a leader signal sequence, which is cleaved off by a signal peptidase in order to yield the processed, mature EPO.
  • signal peptidases recognize only restricted amino acid residues at the P1' and P3' cleavage site (R. E. Dalbey et al. Protein Sci. 6, 1129 (1997).
  • a biologically added protecting peptide has to be built up from an N- terminal amino acid sequence of at least three amino acids for cleavage of the signal sequence, followed by an amino acid sequence for enzymatic or chemical removal of the protecting group. If the recognition sequences of both the signal peptidase and the cleavage protease are identical or closely related, then the sequence of the biologically added protecting group can be reduced to a few amino acids.
  • N-terminal selective modification is obtained by chemoselective ligation to an aldehyde (or ketone)-functionalized target macromolecule (European Patent Application EP 788,375; Gaertner, H F, Offord, R E, Bioconjugate Chem., 7 (1), 38-44 (1996)).
  • this method only works for N-terminal serines or threonines.
  • the Roche's US patent application US 2002/0115833 although discloses the N-terminally pegylated EPO, it specifically defines their formulation of PEG-EPO being pegylated specifically by PEG-NHS (PEG-N-hydroxy succinimide).
  • PEG-NHS PEG-N-hydroxy succinimide
  • the PEG-NHS reacts non-specifically to all lysines, so all lysines needs to be protected by a process called citraconylation.
  • An object of the present invention is to provide a conjugate of Erythropoietin glycoprotein having higher bioactivity.
  • a conjugate comprises an Erythropoietin glycoprotein having N-terminal ⁇ - amino group and aldehyde derivatives of polyethylene glycol, said erythropoietin glycoprotein being selected from the group consisting of human erythropoietin and analogs thereof, said aldehyde derivative of methoxy polyethylene glycol (mPEG) of the formula:
  • R alkyl group of formula -(CH 2 ) n -
  • n 2 to 4.
  • the polyethylene glycol according to the present invention is a mono or branched polyethylene glycol preferably monoPEG and molecular weight of the PEG is about 20 to 60 kda.
  • the R is preferably ethyl, propyl or butyl.
  • Said erythropoietin according to the present invention is a recombinant erythropoietin (rhEPO) produced in animal cells.
  • a process for making the conjugate of the present invention comprises steps of: expressing, fermenting, and purifying rhEPO protein in recombinant animal cells and reacting EPO with PEG-Aldehyde to obtain N- terminally pegylated EPO by reductive amination reaction and then further purified to obtain pure N-terminally pegylated EPO.
  • PEG - Aldehyde added at an about molar ratio of ⁇ 10:2 preferably at 7.5:1 molar ratio of PEG: EPO.
  • the fermentation step according to the present invention is serum free.
  • a pharmaceutical composition comprises the N-pegylated Erythropoietin conjugate of the present invention and a pharmaceutically acceptable excipient.
  • FIG 1 Shows the SDS-PAGE Analysis of native EPO (left lane), PEG- lys-EPO (center lane), and PEG-N-EPO (right lane).
  • FlG 2 Shows the Rat Reticulocyte Count Bioassay with Native EPO, PEG-N-
  • FIG 3 Shows the Quantitation of Pegylated-N-EPO by LC-MS
  • FIG 4 shows the tabular representation of peptides generated by digestion of EPO and PEG-N-EPO with endoproteinase Lys-C.
  • FIG 5 shows the peptide mapping chromatogram of EPO and PEG-N-
  • 0 C refers to degrees Celsius
  • mmol refers to mill mole or mill moles
  • mg refers to milligrams
  • ⁇ g refers to micrograms
  • ml or mL refers to milliliters
  • ⁇ l or ⁇ L refers to micro liters.
  • EPO erythropoietin
  • huEPO erythropoietin
  • EPO is a glycoprotein hormone that is secreted by the human kidney, which stimulates formation of erythrocytes (erythropoiesis) in human bone marrow.
  • the amino acid sequence of the predominant allelic variant of the protein portion of erythropoietin is known.
  • EPO consists of 165 amino acids, is comprised of about 40% carbohydrate, by mass, and has a total molecular weight of approximately 30.6 kDa.
  • the carbohydrate structure of EPO is heterogeneous, whereas the amino acid sequence of the predominant human allelic variant is not. Therefore, these terms refer to a heterogeneous group of EPO or huEPO molecules.
  • PEG-N-EPO means an N-terminally pegylated EPO
  • PEG-Lys-EPO means Lysine pegylated EPO
  • CHO cell means Chinese Hamster Ovary cell.
  • rhEPO means Recombinant Erythropoietin.
  • Erythropoietin compound means a glycosylated protein having nearly the same amino acid sequence as EPO, and having the ability to increase hematocrit when properly administered to a mammal, but differing from EPO in having one or more amino acid modifications.
  • An amino acid modification may be an insertion, a deletion, a replacement, or an inversion of one or more amino acids.
  • Erythropoietic activity refers to the ability of a compound to stimulate erthyropoiesis. Erythropoietic activity can be assessed in vitro, as well as in vivo.
  • Erythropoietic activity generally refers to the ability of a compound to cause an increase in hematocrit levels from an established baseline when administered by an acceptable route of administration at effective doses. In vitro activity can be determined
  • the invention provides a conjugated erythropoietin compound comprising an aldehyde group of PEG molecule primarily attached to N- terminal amino acid and having formula 1 :
  • the conjugate of the present invention can be represented by the formula 2:
  • R alkyl group of formula -(CH 2 )n.
  • the erythropoietin glycoprotein is recombinant EPO being selected from the group consisting of human erythropoietin glycoprotein and analog thereof.
  • the recombinant EPO (rhEPO) produced by growing recombinant human EPO synthesized by recombinant CHO cells is secreted to the culture medium which is then purified by different high performance chromatography steps.
  • the purified EPO thus obtained has similar biological and chemical characteristics as compared to International Standards.
  • Recombinant EPO may be prepared via expression in
  • Expression of proteins, including EPO, by endogenous gene activation is well known in the art and is disclosed, for example in U.S. Pat. Nos. 5,733,761 , 5,641 ,670, 5,733,746, 5,994,122,
  • the preferred EPO species for the preparation of erythropoietin glycoprotein products are human EPO species.
  • the aldehyde derivative of PEG has following formula:
  • R alkyl group of formula -(CHa) n -
  • the polyethylene glycol according to the present invention is a mono or branched polyethylene glycol preferably monoPEG and molecular weight of the PEG is about 20 to 60 kda.
  • the R is preferably ethyl, propyl or butyl or n is 2 to 4.
  • the invention is also directed to a process of preparing a PEG-N- EPO conjugate by expressing, fermenting, and purifying rhEPO protein in recombinant animal cells preferably fermentation is serum free, the reaction of EPO with mPEG-Aldehyde preferably with Propionaldehyde to obtain N-terminally pegylated EPO.
  • EPO reacts with mPEG- propionaldehyde by the reductive amination reaction.
  • the addition of mPEG-propionaldehyde is at about molar ratio of 10:2, preferably at 7.5:1 molar ratio of PEG: EPO.
  • the PEG-N-EPO is further subjected to purification.
  • erythropoietic compound having the PEG - N- Erythropoietin which is an N-terminally pegylated EPO, or erythropoietin analog along with the pharmaceutically acceptable carrier.
  • the pharmaceutically acceptable carriers include adjuvants, excipients, surfactants, and the like that are commonly known and used by the person skilled in the art for the preparation of pharmaceutical compositions.
  • compositions may include various dosage forms including topical, injectable, and the like.
  • a method of treating anemia using erythropoietic compound or a pharmaceutical composition containing the erythropoietic compound of the present invention is provided.
  • the present PEG-N-EPO conjugate Compared to unmodified EPO (i.e., EPO without a PEG attached) and conventional PEG-EPO conjugates, the present PEG-N-EPO conjugate have an increased circulating half-life and plasma residence time, decreased clearance, and increased clinical activity in vivo.
  • the conjugate of this invention have the same uses as EPO.
  • the conjugate of this invention are useful to treat patients by stimulating the division and differentiation of committed erythroid progenitors in the bone marrow in the same way EPO is used to treat patients.
  • conjugate of this invention can be used in the same manner as unmodified EPO.
  • conjugate of this invention have an increased circulating half-life and plasma residence time, decreased clearance, and increased clinical activity in vivo.
  • N-terminal .alpha. -amino group refers to the N-terminal amino residue of a peptide, i.e. that end of a peptide or protein chain having an amino acid with a free alpha-amino (NH. sub.2--) group.
  • the glycoprotein of the conjugate as defined above is a human erythropoietin.
  • Human erythropoietin and analogous proteins as defined above can be expressed by endogenous gene activation.
  • EP-A 0 267 678 discloses an ion exchange chromatography on S-Sepharose, a preparative reverse phase HPLC on a C. sub.8 column and a gel filtration chromatography for the purification of EPO produced in serum-free culture after dialysis.
  • the gel filtration chromatography step can be replaced by ion exchange chromatography on S-Sepharose fast flow. It is also proposed that a dye chromatography on a Blue Trisacryl column be carried out before the ion exchange chromatography.
  • a process for the purification of recombinant EPO is also described by Nobuo, I. et al., J. Biochem. 107 (1990) 352-359.
  • EPO is treated with a solution of Tween.RTM. 20, phenylmethylsulfonyl fluoride, ethylmaleimide, pepstatin A, copper sulfate and oxamic acid prior to the purification steps.
  • EPOsf erythropoietin in a serum free fermentation process
  • PEG-N-EPO is an N-terminally pegylated EPO produced by the following procedure. A 6 mg of EPO was dissolved in 20 mM phosphate buffer containing 150 mM NaCI with pH adjusted at 5.0. The EPO solution was then mixed with mPEG-propionaldehyde 2OK (M.W.: 20,000, manufactured by SunBio, Inc.). The amount of mPEG- propionaldehyde added was 7.5: 1 molar ratio of PEG: EPO. After addition of the PEG, the reaction mixture was stirred well to facilitate the complete dissolution of PEG powder into the EPO solution. After the PEG addition, 100 mM of sodium cyanoborohydride was added and mixed well. The reaction was allowed to continue for 18 hours. In the meanwhile the preparation of the chromatography process was done.
  • chromatography purification is to obtain pure mono- PEG-EPO.
  • HiTrap Q-sepharose chromatography column (Amersham Pharmacia) was packed and equilibrated with 10 mM Tris buffer of pH 8.0. Then the reaction mixture was loaded onto the column, which was then washed with 30 CV (column volume) of 10 mM Tris buffer of pH 8.0 to wash off all remaining free PEG that had not reacted with EPO.
  • the column was eluted with 40 CV of 25 mM NaCI in 10 mM Tris buffer of pH 8.0 to elute di-PEG-EPO (EPO with two PEGs attached to it).
  • the next step was to elute with 40 CV of 65 mM NaCI in 10 mM Tris buffer of pH 8.0 to elute mono-PEG-EPO (EPO with one PEG attached to it).
  • the next step was to elute with 12 CV of 200 mM NaCI in 10 mM
  • Tris buffer of pH 8.0 to elute native EPO which was followed by 8 CV of 1 M NaCI in 10 mM Tris buffer of pH 8.0 to elute any remaining bound material.
  • the collected mono-PEG-EPO fractions were then loaded onto Superdex 200 chromatography (Amersham Pharmacia) to further refine the purity of mono-PEG-EPO.
  • the column was eluted with 20 mM phosphate buffer containing 150 mM NaCI of pH 7.0 to elute the pure mono-PEG-EPO.
  • the collected fractions of mono-PEG-EPO was then concentrated to the final concentration of 65 ⁇ g/ml in buffer of 20 mM phosphate, 150 mM NaCI and pH 7.0.
  • EPO conjugates can be prepared using mPEG-butyraldehyde and mPEG-pentaldehyde.
  • the lysine-52 Pegylated EPO was produced and purified as described in the US patent US 2002/0115833 A1.
  • the result for the peptide mapping Analysis of PEG-N-EPO shows the digestion of native EPO and PEG-N-EPO by lys-C enzyme (Fig. 4) yielded 9 peptide fragments. Of these, 5 peptide fragments, L1 (AA 1- 20), L3 (AA 46-52), L6 (AA 117-140), L7 (AA 141-152), and L9 (156- 166) were quantitatively obtained and analyzed (Fig 3).
  • the peptide mapping analysis of PEG-N-EPO shows that the peptide fragment (L1) containing the N-terminal has been Pegylated.

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Abstract

La présente invention concerne un conjugué d'érythropoïétine à bioactivité supérieure comprenant une glycoprotéine érythropoïétine avec un groupe a - amino et des dérivés aldéhyde de polyéthylène glycol N-terminal, ladite glycoprotéine érythropoïétine étant sélectionnée dans le groupe formé de l'érythropoïétine humaine et de ses analogues, ledit dérivé aldéhyde de méthoxy polyéthylène glycol étant représenté par la formule PEG-R-CHO, dans laquelle R = un groupe alkyle représenté par la formule -(CH2)n dans laquelle n = 2 à 4. La formule du conjugué de la présente invention est PEG-R-CONH-EPO, formule dans laquelle R = un groupe alkyle représenté par la formule -(CH2)n dans laquelle n = 2 à 4. L'érythropoïétine de la présente invention est une érythropoïétine de recombinaison (rhEPO) produite dans des cellules animales. Ledit conjugué d'érythropoïétine est préparé par expression, fermentation, et purification de la protéine rhEPO dans des cellules animales de recombinaison, et par mise en réaction de l'EPO avec PEG-aldéhyde pour obtenir une EPO à PEG N-terminal par une réaction d'amination réductrice, celle-ci étant ensuite purifiée pour obtenir une EPO à PEG N-terminal pure.
PCT/IN2006/000103 2005-03-17 2006-03-16 Conjugue d'erythropoietine peg n-terminal Ceased WO2007010552A2 (fr)

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IN291/MUM/2005 2005-03-17

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Cited By (13)

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WO2009094172A3 (fr) * 2008-01-22 2009-12-10 Araim Pharmaceuticals, Inc. Peptides protecteurs de tissu et analogues peptidiques pour la prévention et le traitement de maladies et de troubles associés à un endommagement tissulaire
US8101706B2 (en) 2008-01-11 2012-01-24 Serina Therapeutics, Inc. Multifunctional forms of polyoxazoline copolymers and drug compositions comprising the same
US8110651B2 (en) 2008-01-11 2012-02-07 Serina Therapeutics, Inc. Multifunctional forms of polyoxazoline copolymers and drug compositions comprising the same
CN102453087A (zh) * 2010-10-22 2012-05-16 深圳赛保尔生物药业有限公司 一种单取代peg-epo的纯化及制备方法
CN102816227A (zh) * 2012-08-30 2012-12-12 深圳赛保尔生物药业有限公司 回收促红细胞生成素的方法
US8673861B2 (en) 2005-08-05 2014-03-18 Araim Pharmaceuticals, Inc. Tissue protective peptides and uses thereof
CN109096483A (zh) * 2017-06-28 2018-12-28 北京键凯科技股份有限公司 分枝型多元甘醇环氧衍生物交联透明质酸钠凝胶及其制备和应用
US10273277B2 (en) 2010-09-14 2019-04-30 Hoffmann-La Roche Inc. Method for purifying PEGylated erythropoietin
WO2019129876A1 (fr) * 2017-12-29 2019-07-04 F. Hoffmann-La Roche Ag Procédé permettant d'obtenir une composition de protéine pegylée
WO2019129878A1 (fr) * 2017-12-29 2019-07-04 F. Hoffmann-La Roche Ag Procédé permettant d'obtenir une composition de protéine pegylée
WO2020096958A1 (fr) * 2018-11-05 2020-05-14 Bristol-Myers Squibb Company Procédé de purification de protéine pegylée
US11518781B2 (en) 2017-12-29 2022-12-06 Hoffmann-La Roche Inc. Process for providing PEGylated protein composition
CN119080909A (zh) * 2024-08-29 2024-12-06 深圳赛保尔生物药业有限公司 负载peg-epo及间充质干细胞的组合物、药物及其制备方法

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CA2352538A1 (fr) * 1998-11-30 2000-06-08 Eli Lilly And Company Composes erythropoietiques
ATE505204T1 (de) * 2000-12-20 2011-04-15 Hoffmann La Roche Konjugate von erythropoietin (epo) mit polyethylenglykol (peg)
WO2004009627A1 (fr) * 2002-07-19 2004-01-29 Cangene Corporation Composes erythropoietiques pegyles
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US8673861B2 (en) 2005-08-05 2014-03-18 Araim Pharmaceuticals, Inc. Tissue protective peptides and uses thereof
US10100096B2 (en) 2005-08-05 2018-10-16 Araim Pharmaceuticals, Inc. Tissue protective peptides and uses thereof
US9340598B2 (en) 2005-08-05 2016-05-17 Araim Pharmaceuticals, Inc. Tissue protective peptides and uses thereof
US8716245B2 (en) 2005-08-05 2014-05-06 Araim Pharmaceuticals, Inc. Tissue protective peptides and uses thereof
US10166294B2 (en) 2008-01-11 2019-01-01 Serina Therapeutics, Inc. Multifunctional forms of polyoxazoline copolymers and drug compositions comprising the same
US9169354B2 (en) 2008-01-11 2015-10-27 Serina Therapeutics, Inc. Multifunctional forms of polyoxazoline copolymers and drug compositions comprising the same
US8501899B2 (en) 2008-01-11 2013-08-06 Serina Therapeutics, Inc. Multifunctional forms of polyoxazoline copolymers and drug compositions comprising the same
US11213588B2 (en) 2008-01-11 2022-01-04 Serina Therapeutics, Inc. Multifunctional forms of polyoxazoline copolymers and drug compositions comprising the same
US11925689B2 (en) 2008-01-11 2024-03-12 Serina Therapeutics, Inc. Multifunctional forms of polyoxazoline copolymers and drug compositions comprising the same
US8110651B2 (en) 2008-01-11 2012-02-07 Serina Therapeutics, Inc. Multifunctional forms of polyoxazoline copolymers and drug compositions comprising the same
US8101706B2 (en) 2008-01-11 2012-01-24 Serina Therapeutics, Inc. Multifunctional forms of polyoxazoline copolymers and drug compositions comprising the same
EP2933264A3 (fr) * 2008-01-22 2015-12-09 Araim Pharmaceuticals, Inc. Peptides protecteurs de tissu et analogues peptidiques pour la prévention et le traitement de maladies et de troubles associés à un endommagement tissulaire
CN102066413B (zh) * 2008-01-22 2015-01-21 阿拉伊姆药品公司 用于预防和治疗组织损伤相关疾病和病症的组织保护肽和肽类似物
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US9580465B2 (en) 2008-01-22 2017-02-28 Araim Pharmaceuticals, Inc. Tissue protective peptides and peptide analogs for preventing and treating diseases and disorders associated with tissue damage
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