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The disulfide exchange can be hs at physiological pH, although the reaction rate is slower. As used herein, “XTEN” specifically excludes whole antibodies or antibody fragments e. In contrast, the low tendency of non-repetitive sequences to aggregate enables the design of long-sequence XTENs with a relatively low frequency of charged amino acids that dps otherwise be likely to aggregate if the sequences were repetitive.
Such polymers can be fractionated by various methods to increase fraction of the target entity in the product. K d is the terminal elimination rate constant calculated by linear regression of the terminal linear portion of the log concentration vs.
An “expression vector” is a polynucleotide which, when introduced into an appropriate host cell, can be transcribed and translated into a. In general, XTEN as monomers or as multimers with cumulative lengths longer that about residues incorporated into the conjugates result in longer half-life compared to shorter cumulative lengths, e.
For example, an atom residue 0401 a payload could be a sulfur residue of a cysteine thiol reactive group in a payload, a nitrogen molecule of an amino reactive group of a peptide or polypeptide or small molecule payload, a carbon or oxygen residue or a reactive carboxyl or aldehyde group of a peptide, protein or a small molecule or synthetic, organic drug.
In one embodiment of the foregoing composition, the composition has the configuration of formula III:.
By “net charge density” of a protein or peptide herein is meant the net charge divided by the total number of amino acids in the protein. Different embodiments of the present invention may have different release gw, resulting in different therapeutic amounts.
WO2013130683A2 – Xten conjugate compositions and methods of making same – Google Patents
In another embodiment, the XTEN-folate conjugate composition has, under physiologic conditions, an apparent molecular weight factor that is about 3 to about 20, or is about 5 to about 15, or is about 8 to about 12, or is about 9 to about 10 relative to the actual molecular weight of the conjugate.
In one embodiment, the repetitiveness of a polypeptide of a predetermined length can be calculated hereinafter “subsequence score” according to the formula given by Equation I:. Acute and repeat-dose toxicity studies of the 6-maleimidocaproyl hydrazone derivative of doxorubicin DOXO-EMCHan albumin-binding prodrug of the anticancer agent doxorubicin. Representative protecting groups for carboxylic acids and hydroxyls include esters such as a p-methoxybenzyl esteramides and hydrazides; for amino groups, carbamates such as tert-butoxycarbonyl and amides; for hydroxyl groups, ethers and esters; for thiol groups, thioethers and thioesters; for carbonyl groups, acetals and ketals; and the like.
In this white paper, we will. A Phase I and pharmacological study of the platinum polymer AP given as an intravenous infusion once every 3 weeks in patients with solid tumors. These repetitive amino 04001b may also tend dxp form contacts resulting in crystalline or pseudocrystaline structures. When we created our character set, the numbers were in order from 0 to 9. An example is a scale developed by Levitt, M, et al.
Isolated polypeptide-encoding nucleic acid molecules therefore are distinguished from the specific polypeptide-encoding nucleic acid molecule 040b1 it exists in natural cells.
Macromolecules can be taken up by the cell either through receptor-mediated endocytosis, adsorptive endocytosis or fluid phase endocytosis Jain R. When conjugated to the XTEN carrier with a cleavable linker, it is cleaved by cathepsin once the conjugate has entered a tumor cell, thus activating the antimitotic mechanism.
In a first aspect, the invention provides XTEN compositions that are substantially homogeneous in length. The apparent molecular weight factor is the ratio between the apparent molecular weight and the actual molecular weight; the latter predicted by adding, based on amino acid composition, the calculated molecular weight of each type of amino acid in the composition or by estimation from comparison to molecular weight standards in an SDS electrophoresis gel.
In one embodiment, the bivalent conjugate comprises a single molecule of a first payload and a single molecule of a second payload linked to the cysteine-lysine-engineered XTEN by nw.
For example, a predetermined polypeptide length of amino acid residues dsl overlapping 9-amino acid subsequences and 3-mer subsequences, but the subsequence score of any given polypeptide will depend on the absolute number of unique subsequences and how frequently each unique subsequence meaning a different amino acid sequence appears in the predetermined length of the sequence.
Folate is a ligand for the cell receptor known as folate receptor.
WOA2 – Xten conjugate compositions and methods of making same – Google Patents
In another embodiment of the dimeric XTEN conjugate, the first and the second XTEN each comprises one or more cysteine residues, and further comprises a first cross-linker conjugated to each cysteine residue of the first XTEN and a second cross-linker conjugated to each cysteine residue of the second XTEN, wherein the first and the second cross- linkers are independently selected from the group consisting of the cross-linkers set forth in Table A “protecting group” wh a moiety that prevents or blocks reaction of a particular chemically reactive functional group in a molecule under certain reaction conditions.
Maleimidopropyloxy succinimide ester and others described above. XTEN release sites can be engineered to be cleaved by various mammalian proteases a.
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For example, an isolated polypeptide- encoding nucleic acid molecule is other than in the form or setting in which it is found in nature. In other embodiments, the invention provides polynucleotides that encode XTEN that can be linked to genes encoding peptide or polypeptide payloads that can be incorporated into expression vectors and incorporated into suitable hosts for the expression and recovery of the subject XTEN.
The hydrodynamic radius of a protein is conferred by its molecular weight as well as by its structure, including shape or compactness. Three molecules of the resulting XTEN are linked via a trifunctional cross-linker comprising three azide functions, resulting in the final XTEN-payload conjugate. The “hydrodynamic radius” of a protein affects its rate of diffusion in aqueous solution as well as its ability to migrate in gels of macromolecules.
The reduced XTEN thus reacts with drug-linker compounds, or cross-linker reagents, with electrophilic functional groups such as maleimide or a- halo carbonyl, according to, for example, the conjugation method of Klussman et al.
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The apparent molecular weight is determined using size exclusion chromatography SEC or similar methods by comparing to globular protein standards, and is measured in “apparent kD” units. In one embodiment of the foregoing method, the cleavage sequence is capable of being cleaved by trypsin and the protease utilized is trypsin.
In addition, a “concentrated”, “separated” or “diluted” polynucleotide, peptide, polypeptide, protein, antibody, or fragments thereof, is distinguishable from its naturally occurring counterpart in that the concentration or number of molecules per volume is generally greater than that of its naturally occurring counterpart. Avoidance of immunogenicity can attribute to, at least in part, a result of the conformational flexibility of XTEN sequences; i. Accordingly, in certain embodiments, the XTEN-payload can be configured with an XTEN such that the resulting conjugate can have a hydrodynamic radius of at least about 5 nm, or at least about 8 nm, or at least about 10 nm, or about 12 nm, or about 15 nm, or about 20 nm, or about 30 nm or more.
Conventional modification methods such as pegylation, adding to the therapeutic agent an antibody fragment or an albumin molecule, suffer from a number of profound drawbacks.
Flip-Flops Operating Manual Ver. An intermediate is produced by reacting XTEN with a trifunctional linker comprising two azide functions and an NHS function followed by the addition of payload A to the thiol group via maleimide chemistry the order of these two steps can be rsp. Batch 1, lane 1: In some embodiments, a selective medium is used which comprises an antibiotic.
The EPL method is based on protein splicing, the process in which a protein undergoes an intramolecular rearrangement resulting in the extrusion of an internal sequence intein and the joining of the lateral sequences exteins.