WO2017049205A2

WO2017049205A2 - Growth hormone formulation

Info

Publication number: WO2017049205A2
Application number: PCT/US2016/052309
Authority: WO
Inventors: Jeffrey L. Cleland
Original assignee: Amunix Operating Inc.
Priority date: 2015-09-18
Filing date: 2016-09-16
Publication date: 2017-03-23
Also published as: WO2017049205A3

Abstract

A stable pharmaceutically acceptable aqueous formulation containing human growth hormone linked to an extended recombinant polypeptide (XTEN) and a buffer is disclosed, wherein the formulation is lacking a surfactant. Also disclosed are associated methods for preparing, storing, and using such formulations.

Description

GROWTH HORMONE FORMULATION

FIELD OF THE INVENTION

The present invention is directed to pharmaceutical formulations containing a fusion protein comprising human growth hormone (hGH) and to methods for making and using such formulations. More particularly, this invention relates to liquid hGH formulations with increased stability.

BACKGROUND OF THE INVENTION

Native human growth hormone (hGH) in its most prominent mature form of pituitary

(pit)-hGH is a single polypeptide chain protein consisting of 191 amino acids. The protein is internally cross-linked by two disulphide bridges and in monomelic form has a molecular weight of 22 kDa. Growth hormone (GH) of animal species is closely homologous in amino acid sequence to that of humans and is therefore very similar in its characteristics. A major biological effect of GH is to promote growth throughout a range of organs and tissues in the body. GH responsive organs or tissues include the liver, intestine, kidneys, muscles, connective tissue and the skeleton. For a more detailed review of the stability and characterization of hGH, see Pearlman and Bewley, Chapter 1 in Stability and

Characterization of Protein and Peptide Drugs: Case Histories (Plenum Press, New York, 1993).

Methods for the recombinant production of recombinant hGH (rhGH) are described in U.S. Patent No. 4,601,980 and Goeddel et al, Nature 1979, 281(5732):544-8.

Recombinant hGH (rhGH) is used as a therapeutic and has been approved for the treatment of a number of indications. rhGH is currently sold in a variety of forms worldwide including capsules, syrups, sprays, tablets, powders, vials, and injections. Numerous injectable products are currently on the market in the United States: for instance,

Humatrope™ (Eli Lilly & Co.), Nutropin™ (Genentech), Norditropin™ (Novo-Nordisk), Omnitrope™ (Sandoz/Novartis), Genotropin™ (Pfizer), Saizen/Serostim™ (Serono), Tev- Tropin™ (Teva), and Zorbtive™ (EMD Serono). hGH deficiency leads to dwarfism, for example, which has been successfully treated for decades by exogenous administration of the hormone. In addition to hGH deficiency, hGH has also been approved for the treatment of several other conditions including renal failure (in children), Turner's Syndrome, and cachexia in AIDS patients. The Food and Drug Administration (FDA) has approved hGH for the treatment of non-GH-dependent short stature and hGH has been investigated for the treatment of a variety of physiological conditions, including aging, frailty in the elderly, short bowel syndrome, and congestive heart failure. Target populations for hGH treatment include children with idiopathic short stature (ISS) and adults with GHD-like symptoms.

A long appreciated problem with aqueous liquid formulations of pharmaceutical proteins, not just hGH, has been that of instability during storage over a period of time.

While liquid hGH formulations are described in the patent literature (see, e.g. U.S. Patent Nos. 5,763,394; 7,662,393; 8,338,374 and 8,409,586), many of the major injectable hGH products are formulated as lyophilized preparations requiring reconstitution prior to use. For example, in the United States, Genotropin™ lyophilized powder is provided in a 5 mg or 12 mg two-chamber cartridge to be reconstituted with water for injection

(www.pfizermedicalinformation.com/en-us/genotropin; accessed September 3, 2015);

Humatrope™ lyophilized powder is provided in a 5 mg vial or 6-24 mg cartridge to be reconstituted with water for injection (pi.lilly.com/us/humatrope-pi.pdf; accessed September 3, 2015).

Thus, there remains a need to develop aqueous liquid formulations of hGH that do not require reconstitution and are chemically stable for an extended period, which have the advantages of eliminating reconstitution errors, thereby increasing dosing accuracy, as well as simplifying the use of the product clinically, thereby increasing patient compliance. Such formulations also allow for improvements in the design and use of pre-filled syringes or other devices, which are sold to patients containing pre-mixed liquid formulations of rhGH (e.g., Norditropin™). Therapeutic aqueous liquid formulations of hGH preferably should be stable for 2-3 years at refrigerated temperatures and for several weeks near room temperature. For example, prior to initial use, the refrigerated shelf-life of the Norditropin FlexPro pre-filled pen is 2 years

(www. novonordi sk. com/ content/ dam/Denmark/HQ/hcp/Documents/Norditropin%20FlexPro- SmPC_August2012.pdf; accessed September 3, 2015). However, a major challenge facing the development of such viable aqueous liquid formulations of hGH is that hGH in aqueous solution is known to undergo a variety of degradative changes.

Human growth hormone degrades by three primary pathways: deamidation, oxidation and aggregation (Pearlman and Bewley, supra at 26-42; Cleland et al., Crit Rev Ther Drug Carrier Syst. 1993; 10(4):307-77). Deamidation is minimized between pH 5 and 6 and more preferably near pH 5.5 (Pearlman and Bewley, supra at 27-29). However, solubility of hGH decreases substantially at this pH because the pi of hGH is 5.2 {Id at 17). Formulations have therefore been developed to balance solubility and deamidation. Agitation induced aggregation of hGH occurs due to the air-water interface denaturation of hGH (Pearlman and Bewley, supra at 36-40; Bam et al, Journal of Pharmaceutical Sciences Vol. 87, No. 12, December 1998). To inhibit aggregation in hGH formulation, surfactants have been added and have been shown to bind to hGH blocking aggregation (Bam et al., Pharmaceutical Research, Vol. 12, No. 1, 1995). These surfactants (e.g. polysorbate 20, polyethylene glycol (PEG)) generate free radical oxygen upon storage causing oxidation of exposed methionine residues in hGH. Overall, hGH formulations have been developed to balance the degradation pathways to enable sufficient stability for commercial application (e.g., 2 years at refrigerated conditions).

This invention provides a highly stable aqueous liquid formulation of a fusion protein containing rhGH. In particular, the invention provides a liquid formulation of a rhGH-XTEN fusion protein that exhibits excellent long-term stability without the use of a surfactant.

SUMMARY OF THE INVENTION

A stable pharmaceutically acceptable aqueous formulation containing a fusion protein comprising human growth hormone linked to an extended recombinant polypeptide (XTEN) and a buffer is disclosed, wherein the formulation is lacking a surfactant. Also disclosed are associated methods for preparing, storing, and using such formulations.

Aspects of the invention include a storage stable aqueous liquid formulation comprising (i) about 25 mg/ml to about 150 mg/ml of a fusion protein comprising an extended recombinant polypeptide (XTEN) fused to a growth hormone (GH) sequence, wherein the GH sequence is at least 90% identical to the amino acid sequence of SEQ ID NO:43, and (ii) a buffer; characterized in that the aqueous liquid formulation is at a pH of about 4 to about 6, in the absence of a surfactant. In some embodiments, the aqueous liquid formulation is characterized in that it is free of a preservative. In some embodiments, the aqueous liquid formulation is characterized in that the pH is about 5 to about 6. In some embodiments, the aqueous liquid formulation is characterized in that the pH is 5.2 to 5.8. In some embodiments, the aqueous liquid formulation is characterized in that the pH is about 5.5. In some embodiments, the aqueous liquid formulation is characterized in that the buffer is selected from the group consisting of histidine, citrate, and succinate buffers. In some embodiments, the aqueous liquid formulation is characterized in that the buffer is a histidine buffer at a concentration of about 20 mM.

In some embodiments, the aqueous liquid formulation is characterized in that the concentration of the fusion protein is selected from the group consisting of about 25 mg/ml, about 50 mg/ml, about 75 mg/ml, about 100 mg/ml, about 125 mg/ml, and about 150 mg/ml. In some embodiments, the aqueous liquid formulation is characterized in that the

concentration of the fusion protein is about 50 mg/ml. In some embodiments, the aqueous liquid formulation is characterized in that the concentration of the fusion protein is about 100 mg/ml. In some embodiments, the aqueous liquid formulation is characterized in that the concentration of the fusion protein is about 150 mg/ml.

In some embodiments, the aqueous liquid formulation is characterized in that the aqueous liquid formulation is storage stable for at least about one month at about 23±3°C. In some embodiments, the aqueous liquid formulation is characterized in that the aqueous liquid formulation is storage stable for at least about 6 months to about 3 years at about 5±3 °C. In some embodiments, the aqueous liquid formulation is characterized in that the aqueous liquid formulation is storage stable for at least about 6 months to about 18 months at about 5±3 °C. In some embodiments, the aqueous liquid formulation is characterized in that the aqueous liquid formulation is storage stable at about 5±3 °C for at least about 3 months, for at least about 6 months, for at least about 9 months, for at least about 12 months, for at least about 18 months, for at least about 21 months, for at least about 24 months, for at least about 27 months, for at least about 30 months, for at least about 36 months, or for at least about 42 months.

In some embodiments, the aqueous liquid formulation is characterized in that the oxidation of the fusion protein is reduced as compared to the oxidation of the corresponding fusion protein formulated with a surfactant. In some embodiments, the aqueous liquid formulation is characterized in that the deamidation of the fusion protein is reduced as compared to the deamidation of the corresponding fusion protein formulated at a higher pH. In some embodiments, the aqueous liquid formulation is characterized in that the isolated fusion protein does not show detectable oxidation or deamidation following incubation for about 8 hours at about 40 °C.

In some embodiments, the aqueous liquid formulation is characterized in that it further comprises a tonicifying agent. In some embodiments, the aqueous liquid formulation is characterized in that the tonicifying agent is NaCl. In some embodiments, the aqueous liquid formulation is characterized as having an osmolality between about 300 mOsm to about 600 mOsm.

In some embodiments, the aqueous liquid formulation is characterized in that the formulation is disposed in a container containing at least one dose. In some embodiments, the container comprises a syringe. In some embodiment, the container comprises a glass cartridge. In some embodiments, the container comprises a vial.

In some embodiments, the aqueous liquid formulation is characterized in that it is for injection into a subject with a needle. In some embodiments, the needle size is selected from the group consisting of 26 gauge, 27 gauge, 29 gauge, and 30 gauge.

In some embodiments, the aqueous liquid formulation is characterized in that it comprises the fusion protein according to the formula I:

(XTEN l )_x-GH-(XTEN_2)y

wherein independently for each occurrence:

(a) x is either 0 or 1; and

(b) y is either 0 or 1, wherein x+y>l;

wherein XTEN l and XTEN 2 comprise identical or different sequences. In some embodiments, the aqueous liquid formulation is characterized in that the XTEN l comprises a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOS: 18-34, 38, 39, 41, and 42. In some embodiments, the aqueous liquid formulation is characterized in that the XTEN_2 comprises a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOS: 13-17, 35, 36, 37, and 40. In some embodiments, the aqueous liquid formulation is characterized in that the XTEN l comprises SEQ ID NO:39. In some embodiments, the aqueous liquid formulation is characterized in that the XTEN 2 comprises SEQ ID NO: 35. In some embodiments, the aqueous liquid formulation is characterizes in that XTEN l comprises SEQ ID NO: 39 and XTEN 2 comprises SEQ ID NO: 35. In some embodiments, the aqueous liquid formulation is characterized in that the fusion protein sequence is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 1. In some embodiments, the aqueous liquid formulation is characterized in that the fusion protein comprises the amino acid sequence of SEQ ID NO: 1. Some aspects of the invention relate to a method of making a storage stable aqueous liquid formulation of a fusion protein comprising an extended recombinant polypeptide (XTEN) linked to a growth hormone (GH) sequence, wherein the GH sequence is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO:43, comprising mixing said fusion protein and an aqueous, pharmaceutically acceptable vehicle which includes i) said fusion protein at a concentration of about 25 mg/ml to about 150 mg/ml; and ii) a buffer providing a pH of about 4 to about 6; wherein said aqueous liquid formulation is free of a surfactant. In some embodiments, the method is characterized in that it is formulated without use of a preservative.

Some aspects of the invention relate to a method of treating a growth-hormone related condition in a subject, comprising administering to the subject a therapeutically effective amount of the aqueous liquid formulation according to the presently described invention, wherein the growth-hormone related condition is selected from the group consisting of growth-hormone deficiency, Turner's Syndrome, Prader-Willi Syndrome, small for gestational age (SGA), idiopathic short stature, AIDS wasting, multiple sclerosis, Crohn's disease, ulcerative colitis, and muscular dystrophy. In some embodiments, the method is characterized in that the therapeutically effective amount is administered at least 48 h, or at least 72 h, or at least about 96 h, or at least about 120 h, or at least about 7 days, or at least about 14 days, at least about 21 days, or at least about 30 days between consecutive doses. In some embodiments, the method is characterized in that the growth-hormone related condition is growth-hormone deficiency. In some embodiments, the method is characterized in that the growth-hormone related condition is Turner's Syndrome. In some embodiments, the method the method is characterized in that the growth-hormone related condition is Prader-Willi Syndrome. In some embodiments, the method is characterized in that the growth-hormone related condition is idiopathic short stature. In some embodiments, the method is

characterized in that the growth-hormone related condition is small for gestational age (SGA). In some embodiments, the method is characterized in that the growth-hormone related condition is selected from the group consisting of AIDS wasting, multiple sclerosis, Crohn' s disease, ulcerative colitis, and muscular dystrophy.

Some aspects of the invention relate to the use of a therapeutically effective amount of the aqueous liquid formulation of according to the presently described invention in the preparation of a medicament for treating a growth-hormone related condition, wherein the growth-hormone related condition is selected from the group consisting of growth-hormone deficiency, Turner's Syndrome, Prader-Willi Syndrome, small for gestational age (SGA), idiopathic short stature, AIDS wasting, multiple sclerosis, Crohn's disease, ulcerative colitis, and muscular dystrophy.

In some embodiments, the aqueous liquid formulation is characterized in that it is for use in treating a growth-hormone related condition, wherein the growth-hormone related condition is selected from the group consisting of growth-hormone deficiency, Turner's Syndrome, Prader-Willi Syndrome, small for gestational age (SGA), idiopathic short stature, AIDS wasting, multiple sclerosis, Crohn's disease, ulcerative colitis, and muscular dystrophy.

INCORPORATION BY REFERENCE

All publications, patent and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application were specifically and individually indicated to be incorporated by reference.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 provides the amino acid sequence of an hGH-XTEN fusion protein designated VRS-317 (hGH sequence is underlined and shown in bold) (SEQ ID NO: 1).

FIG. 2 is a strong anion exchange chromatogram of an aqueous formulation according to the present invention showing degradation of VRS-317 at 40°C via hydrolysis, or truncation, of the XTEN polypeptide in VRS-317.

FIG. 3 is a strong anion exchange chromatogram of an aqueous formulation according to the present invention containing different concentrations of VRS-317 showing that VRS- 317 in the formulation is not degraded.

FIG. 4. is a size exclusion chromatogram of an aqueous formulation according to the present invention containing different concentrations of VRS-317 showing that VRS-317 in the formulation is not aggregated.

FIG. 5. is a size exclusion chromatogram of an aqueous formulation according to the present invention containing different concentrations of VRS-317, showing that VRS-317 in the formulation is not aggregated after passing through a 30 gauge needle.

FIG. 6 is a reverse phase chromatogram of tryptic fragments of control VRS-317 (black) or VRS-317 treated with hydrogen peroxide for 72 hours (gray). Inset: A & B are expanded views of the parts of the chromatogram (underlined) where degradation of VRS- 317 was observed.

FIG. 7 is a reverse phase chromatogram of tryptic fragments of control VRS-317 (black) or VRS-317 stressed with elevated pH and temperatures (gray). Inset: An expanded view of the reverse phase chromatogram, highlighting the observed degradation.

FIG. 8 shows SE-HPLC results comparing the degree of VRS-317 aggregation seen in aqueous formulations according to the present invention in the presence or absence of surfactant, following 4 hour agitation stress experiments at ambient temperature.

FIG. 9 shows graphs of integrated percent peak areas from the size exclusion chromatogram of an aqueous formulation according to the present invention containing VRS- 317 sampled periodically over up to 24 months of storage at 5°C or 25°C.

FIG. 10 shows a graph of integrated percent peak areas from the reverse phase chromatogram of tryptic fragments of an aqueous formulation according to the present invention containing VRS-317 sampled over 1 month of storage at 25°C.

FIG. 11 shows a graph of pH readings taken periodically over up to 24 months of an aqueous formulation according to the present invention containing VRS-317 stored at 5°C or 25°C.

DETAILED DESCRIPTION OF THE INVENTION

I) DEFINITIONS

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood to one of ordinary skill in the art to which the inventions described herein belong. For purposes of interpreting this specification, the following definitions will apply and whenever appropriate, terms used in the singular will also include the plural and vice versa. In the event that any definition set forth conflicts with any document incorporated herein by reference, the definition set forth below shall control.

The terms "polypeptide", "peptide", and "protein" are used interchangeably herein to refer to polymers of amino acids of any length. The polymer may be linear or branched, it may comprise modified amino acids, and it may be interrupted by non-amino acids. The terms also encompass an amino acid polymer that has been modified, for example, by disulfide bond formation, glycosylation, lipidation, acetylation, phosphorylation, or any other manipulation, such as conjugation with a labeling component. "Growth Hormone" or "GH" means a growth hormone polypeptide, including, without limitation, native sequence human and non-human GH, and amino acid sequence variants thereof having growth hormone biological activity. The term "native sequence" specifically encompasses naturally occurring truncated or variant forms, including alternatively spliced forms and naturally occurring allelic variants. A non-human grown hormone can be of any non-human animal species, such as a non-human mammalian species. The term "native sequence human growth hormone (hGH)" or "native hGH" includes, but is not limited to, the hGH originally derived from the pituitary gland having a molecular weight of about 22, 129 daltons (22kD hGH) and a naturally occurring variant having a molecular weight of about 20,000 daltons (20kD hGH). The 20kD hGH has an amino acid sequence that corresponds to that of 22kD hGH consisting of 191 amino acids except that 15 amino acid residues from the 32nd to the 46th of 22kD hGH are missing. Some reports have shown that the 20kD hGH has been found to exhibit lower risks and higher activity than 22kD hGH. The invention contemplates use of the 22 kD, the 20kD hGH, as well as species and sequence variants and truncated fragments thereof as being appropriate for use as a fusion partner with XTEN disclosed herein for hGH-XTEN compositions. The cloned gene for hGH has been expressed in a secreted form in Escherichia coli (United States Patent No. 4,898,830; Chang, C. N., et al., Gene 55: 189 [1987]) and its DNA and amino acid sequence has been reported (Goeddel, et al. Nature ,281 :544 [1979]); Gray, et al., Gene 39: 247[1985]). The terms "GH" and "hGH" specifically include recombinant human growth hormone (rhGH) produced in any recombinant host cell, including prokaryotic and eukaryotic hosts.

The term "storage stable", as it relates to a pharmaceutical liquid formulation comprising hGH fusion protein, is a formulation that results in less than 5% aggregation of hGH fusion protein, less than 15% deamidation, or less than 10% oxidation of hGH fusion protein when such pharmaceutical liquid formulation comprising hGH is stored at 2° C to 8° C. In one embodiment, such liquid formulation comprising hGH is long-term stable for at least 2-3 years, and in an alternate embodiment, such liquid formulation comprising hGH is storage stable for 24 months or for 6 to 18 months, or stable at least 12 months. As will be obvious to one skilled in the art, the long-term stability of hGH may be determined directly by incubating the formulations for the above noted times, or the long-term stability may be predicted by the methods described herein, or other known methods.

As used herein the term "amino acid" refers to either natural and/or unnatural or synthetic amino acids, including but not limited to glycine and both the D or L optical isomers, and amino acid analogs and peptidomimetics. Standard single or three letter codes are used to designate amino acids.

A "fragment" is a truncated form of a native biologically active protein that retains at least a portion of the therapeutic and/or biological activity. A "variant" is a protein with sequence homology to the native biologically active protein that retains at least a portion of the therapeutic and/or biological activity of the biologically active protein. For example, a variant protein may share at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% amino acid sequence identity with the reference biologically active protein. As used herein, the term "biologically active protein moiety" includes proteins modified deliberately, as for example, by site directed mutagenesis, insertions, or accidentally through mutations. Thus, "GH variant," "hGH variant" or "rhGH variant" comprises an amino acid sequence which differs from that of a native sequence GH, hGH or rhGH by virtue of at least one amino acid modification, preferably one or more amino acid substitution(s). Preferably, the variant has at least one amino acid substitution compared to a native sequence polypeptide, e.g. from about one to about ten amino acid substitutions, and preferably from about one to about five amino acid substitutions in the sequence of the parent polypeptide.

The term "natural L-amino acid" means the L optical isomer forms of glycine (G), proline (P), alanine (A), valine (V), leucine (L), isoleucine (I), methionine (M), cysteine (C), phenylalanine (F), tyrosine (Y), tryptophan (W), histidine (H), lysine (K), arginine (R), glutamine (Q), asparagine (N), glutamic acid (E), aspartic acid (D), serine (S), and threonine (T).

The term "non-naturally occurring," as applied to sequences and as used herein, means polypeptide or polynucleotide sequences that do not have a counterpart to, are not complementary to, or do not have a high degree of homology with a wild-type or naturally- occurring sequence found in a mammal. For example, a non-naturally occurring polypeptide or fragment may share no more than 99%, 98%, 95%, 90%, 80%, 70%, 60%, 50% or even less amino acid sequence identity as compared to a natural sequence when suitably aligned.

The terms "hydrophilic" and "hydrophobic" refer to the degree of affinity that a substance has with water. A hydrophilic substance has a strong affinity for water, tending to dissolve in, mix with, or be wetted by water, while a hydrophobic substance substantially lacks affinity for water, tending to repel and not absorb water and tending not to dissolve in or mix with or be wetted by water. Amino acids can be characterized based on their hydrophobicity. A number of scales have been developed. An example is a scale developed by Levitt, M, et al, J Mol Biol (1976) 104:59, which is listed in Hopp, TP, et al, Proc Natl Acad Sci U S A (1981) 78:3824. Examples of "hydrophilic amino acids" are arginine, lysine, threonine, alanine, asparagine, and glutamine. Of particular interest are the hydrophilic amino acids aspartate, glutamate, and serine, and glycine. Examples of

"hydrophobic amino acids" are tryptophan, tyrosine, phenylalanine, methionine, leucine, isoleucine, and valine.

The term "hGH-XTEN", as used herein, is meant to encompass fusion polypeptides that comprise a payload region comprising a biologically active GH that mediates one or more biological or therapeutic activities associated with growth hormone and at least one other region comprising at least a first XTEN polypeptide that serves as a carrier. In one embodiment, the invention provides an hGH-XTEN fusion protein comprising a sequence set forth in Table 2. In a preferred embodiment, the invention provides an hGH-XTEN fusion protein comprising SEQ ID NO: 35 and SEQ ID NO: 39.

The term "buffer" or "physiologically-acceptable buffer" refers to solutions of compounds that are known to be safe for pharmaceutical or veterinary use in formulations and that have the effect of maintaining or controlling the pH of a formulation in the pH range desired for the formulation. Buffers typically involve a weak acid or alkali together with one of its salts. Acceptable buffers for controlling pH at a moderately acidic pH to a moderately basic pH include, but are not limited to, such compounds as phosphate, acetate, citrate, arginine, TRIS, and histidine. "TRIS" refers to 2-amino-2-hydroxymethyl-l, 3, -propanediol, and to any pharmacologically acceptable salt thereof.

"Pharmaceutically acceptable" excipients (vehicles, additives) are those commonly used in the art which can reasonably be administered to a subject mammal to provide an effective dose of the active ingredient employed. For parenteral administration (e.g.

subcutaneous or intramuscular, or topical administration) formulations may be converted into a solution, gel or emulsion, if desired, using the pharmaceutical substances customary for this purpose, such as solubilizers, thickening agents, emulsifiers, agents for tonicity, preservatives or other auxiliaries.

As used herein, the term "surfactant" is used to refer to a surface-active agent, typically a nonionic surfactant, understood and employed by those skilled in the art.

Examples of suitable surfactants include polysorbate (for example, polysorbate 20 and polysorbate 80); poloxamer (e.g. poloxamer 188); Triton; sodium dodecyl sulfate (SDS); sodium laurel sulfate; sodium octyl glycoside; lauryl-, myristyl-, linoleyl-, or stearyl- sulfobetaine; lauryl-, myristyl-, linoleyl- or stearyl-sarcosine; linoleyl-, myristyl-, or cetyl- betaine; lauroamidopropyl-, cocamidopropyl-, linoleamidopropyl-, myristamidopropyl-, palmidopropyl-, or isostearamidopropyl-betaine (e.g. lauroamidopropyl); myristamidopropyl- , palmidopropyl-, or isostearamidopropyl-dimethylamine; sodium methyl cocoyl-, or disodium methyl oleyl-taurate; polyethyl glycol, polypropyl glycol, and copolymers of ethylene and propylene glycol (e.g. Pluronics, PF68 etc).

A "sugar," or "saccharide," or "sugar alcohol," or "polyol" as used herein refers to a substances well known in the art with multiple hydroxyl groups, and includes reducing sugars, nonreducing sugars, sugar alcohols and sugar acids. A "reducing sugar" is one which contains a hemiacetal group that can reduce metal ions or react covalently with lysine and other amino groups in proteins and a "nonreducing sugar" is one which does not have these properties of a reducing sugar. Examples of reducing sugars are fructose, mannose, maltose, lactose, arabinose, xylose, ribose, rhamnose, galactose and glucose. Nonreducing sugars include sucrose, trehalose, stachyose, sorbose, melezitose and raffinose. Mannitol, xylitol, erythritol, threitol, sorbitol, ribitol, myoinositol, galactitol and glycerol are examples of sugar alcohols. As to sugar acids, these include L-gluconate and metallic salts thereof. Where it is desired that the formulation is freeze-thaw stable, the polyol is preferably one which does not crystallize at freezing temperatures (e.g. -20°C.) such that it destabilizes the fusion protein composition in the formulation. Polyols, including mixtures of polyols, can also be used as lyoprotectants in the formulations of the present invention, which protects a protein, e.g. an fusion protein, from damage resulting from lyophilization. Other stabilizing agents, such as metal chelating agents, may also be useful in the invention which are generally known in the art.

A "preservative" is understood in the art to be a compound which can be included in the formulation to substantially reduce bacterial action therein, thus facilitating the production of a multi-use formulation, for example. Examples of preservatives include octadecyldimethylbenzyl ammonium chloride, hexamethonium chloride, benzalkonium chloride (a mixture of alkylbenzyldimethylammonium chlorides in which the alkyl groups are long-chain compounds), and benzelthonium chloride. Other types of preservatives include aromatic alcohols such as phenol, butyl and benzyl alcohol, alkyl parabens such as methyl or propyl paraben, catechol, resorcinol, cyclohexanol, 3-pentanol, and m-cresol.

"Conjugated", "linked," "fused," and "fusion" are used interchangeably herein. These terms refer to the joining together of two or more chemical elements or components, by whatever means including chemical conjugation or recombinant means. For example, a promoter or enhancer is operably linked to a coding sequence if it affects the transcription of the sequence. Generally, "operably linked" means that the DNA sequences being linked are contiguous, and in reading phase or in-frame. An "in-frame fusion" refers to the joining of two or more open reading frames (ORFs) to form a continuous longer ORF, in a manner that maintains the correct reading frame of the original ORFs. Thus, the resulting recombinant fusion protein is a single protein containing two or more segments that correspond to polypeptides encoded by the original ORFs (which segments are not normally so joined in nature).

In the context of polypeptides, a "linear sequence" or a "sequence" is an order of amino acids in a polypeptide in an amino to carboxyl terminus direction in which residues that neighbor each other in the sequence are contiguous in the primary structure of the polypeptide. A "partial sequence" is a linear sequence of part of a polypeptide that is known to comprise additional residues in one or both directions.

"Recombinant" as applied to a polynucleotide means that the polynucleotide is the product of various combinations of in vitro cloning, restriction and/or ligation steps, and other procedures that result in a construct that can potentially be expressed in a host cell.

The terms "percent identity" and "% identity," as applied to polynucleotide sequences, refer to the percentage of residue matches between at least two polynucleotide sequences aligned using a standardized algorithm. Such an algorithm may insert, in a standardized and reproducible way, gaps in the sequences being compared in order to optimize alignment between two sequences, and therefore achieve a more meaningful comparison of the two sequences. Percent identity may be measured over the length of an entire defined polynucleotide sequence, or may be measured over a shorter length, for example, over the length of a fragment taken from a larger, defined polynucleotide sequence, for instance, a fragment of at least 45, at least 60, at least 90, at least 120, at least 150, at least 210 or at least 450 contiguous residues. Such lengths are exemplary only, and it is understood that any fragment length supported by the sequences shown herein, in the tables, figures or Sequence Listing, may be used to describe a length over which percentage identity may be measured.

"Percent (%) amino acid sequence identity," with respect to the polypeptide sequences identified herein, is defined as the percentage of amino acid residues in a query sequence that are identical with the amino acid residues of a second, reference polypeptide sequence or a portion thereof, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity, and not considering any conservative substitutions as part of the sequence identity. Alignment for purposes of determining percent amino acid sequence identity can be achieved in various ways that are within the skill in the art, for instance, using publicly available computer software such as BLAST, BLAST-2, ALIGN or Megalign (DNASTAR) software. Those skilled in the art can determine appropriate parameters for measuring alignment, including any algorithms needed to achieve maximal alignment over the full length of the sequences being compared. Percent identity may be measured over the length of an entire defined polypeptide sequence, or may be measured over a shorter length, for example, over the length of a fragment taken from a larger, defined polypeptide sequence, for instance, a fragment of at least 15, at least 20, at least 30, at least 40, at least 50, at least 70 or at least 150 contiguous residues. Such lengths are exemplary only, and it is understood that any fragment length supported by the sequences shown herein, in the tables, figures or Sequence Listing, may be used to describe a length over which percentage identity may be measured.

The term "non-repetitiveness" as used herein in the context of a polypeptide refers to a lack or limited degree of internal homology in a peptide or polypeptide sequence. The term "substantially non-repetitive" can mean, for example, that there are few or no instances of four contiguous amino acids in the sequence that are identical amino acid types or that the polypeptide has a subsequence score (defined infra) of 10 or less or that there isn't a pattern in the order, from N- to C-terminus, of the sequence motifs that constitute the polypeptide sequence. The term "repetitiveness" as used herein in the context of a polypeptide refers to the degree of internal homology in a peptide or polypeptide sequence. In contrast, a

"repetitive" sequence may contain multiple identical copies of short amino acid sequences. For instance, a polypeptide sequence of interest may be divided into n-mer sequences and the number of identical sequences can be counted. Highly repetitive sequences contain a large fraction of identical sequences while non-repetitive sequences contain few identical sequences. In the context of a polypeptide, a sequence can contain multiple copies of shorter sequences of defined or variable length, or motifs, in which the motifs themselves have non- repetitive sequences, rendering the full-length polypeptide substantially non-repetitive. The length of polypeptide within which the non-repetitiveness is measured can vary from 3 amino acids to about 200 amino acids, about from 6 to about 50 amino acids, or from about 9 to about 14 amino acids. "Repetitiveness" used in the context of polynucleotide sequences refers to the degree of internal homology in the sequence such as, for example, the frequency of identical nucleotide sequences of a given length. Repetitiveness can, for example, be measured by analyzing the frequency of identical sequences.

As used herein, "treatment" or "treating," or "palliating" or "ameliorating" is used interchangeably herein. These terms refer to an approach for obtaining beneficial or desired results including but not limited to a therapeutic benefit and/or a prophylactic benefit. By therapeutic benefit is meant eradication or amelioration of the underlying disorder being treated. Also, a therapeutic benefit is achieved with the eradication or amelioration of one or more of the physiological symptoms associated with the underlying disorder such that an improvement is observed in the pediatric subject, notwithstanding that the subject may still be afflicted with the underlying disorder. The therapeutic benefits may include sustained or normalized parameters such as, but not limited to, annual height velocity (cm/yr) in the first year (12 months) of treatment, IGF-I standard deviation score (IGF-I SDS), height standard deviation score, body weight, body mass index bone age, and pubertal staging. A preferred therapeutic benefit is sustained or normalized annual height velocity (cm/yr) in the first year (12 months) of treatment. For prophylactic benefit, the compositions may be administered to a pediatric subject at risk of developing a particular disease, or to a pediatric subject reporting one or more of the physiological symptoms of a disease, even though a diagnosis of this disease may not have been made. The prophylactic benefits may include sustained or normalized parameters such as, but not limited to, annual height velocity (cm/yr) in the first year (12 months) of treatment, IGF-I standard deviation score (IGF-I SDS), height standard deviation score, body weight, body mass index bone age, and pubertal staging. A preferred prophylactic benefit is sustained or normalized annual height velocity (cm/yr) in the first year (12 months) of treatment.

A "therapeutic effect", as used herein, refers to a physiologic effect, including but not limited to the cure, mitigation, amelioration, or prevention of disease in humans or other animals, or to otherwise enhance physical or mental wellbeing of humans or animals, caused by a fusion polypeptide of the invention other than the ability to induce the production of an antibody against an antigenic epitope possessed by the biologically active protein.

Determination of a therapeutically effective amount is well within the capability of those skilled in the art, especially in light of the detailed disclosure provided herein. The therapeutic effect may include sustained or normalized parameters such as, but not limited to, annual height velocity (cm/yr) in the first year (12 months) of treatment, IGF-I standard deviation score (IGF-I SDS), height standard deviation score, body weight, body mass index bone age, and pubertal staging. A preferred therapeutic effect is sustained or normalized annual height velocity (cm/yr) in the first year (12 months) of treatment.

The terms "therapeutically effective amount" and "therapeutically effective dose" and "pharmaceutically effective amount", as used herein, refers to an amount of a biologically active protein, either alone or as a part of a fusion protein composition, that is capable of having any detectable, beneficial effect on any symptom, aspect, measured parameter or characteristics of a disease state or condition when administered in one or repeated doses to a pediatric subject. Such effect need not be absolute to be beneficial.

II). GENERAL TECHNIQUES

The practice of the present invention employs, unless otherwise indicated,

conventional techniques of immunology, biochemistry, chemistry, molecular biology, microbiology, cell biology, genomics and recombinant DNA, which are within the skill of the art. See Sambrook, J. et al., "Molecular Cloning: A Laboratory Manual," 3rd edition, Cold Spring Harbor Laboratory Press, 2001; "Current protocols in molecular biology", F. M.

Ausubel, et al. eds.,1987; the series "Methods in Enzymology," Academic Press, San Diego, CA.; "PCR 2: a practical approach", M.J. MacPherson, B.D. Hames and G.R. Taylor eds., Oxford University Press, 1995; "Antibodies, a laboratory manual" Harlow, E. and Lane, D. eds., Cold Spring Harbor Laboratory, 1988; "Goodman & Gilman's The Pharmacological Basis of Therapeutics," 11th Edition, McGraw-Hill, 2005; and Freshney, R.I., "Culture of Animal Cells: A Manual of Basic Technique," 4th edition, John Wiley & Sons, Somerset, NJ, 2000, the contents of which are incorporated in their entirety herein by reference. III). hGH FORMULATIONS

The present invention is based, at least in part, on the finding that hGH covalently fused to extended recombinant polypeptides ("XTE " or "XTENs") is highly soluble and stable in aqueous liquid solution at a relatively low pH and in the absence of a surfactant. Specifically, liquid aqueous formulations of hGH-XTEN according to the invention remain stable during storage over a period of at least about 24 months and longer when stored refrigerated (5±3°C) or stable during storage over a period of at least about 1 month and longer when stored at 25±3 °C. According to the present invention, hGH-XTEN can be formulated as a storage stable aqueous liquid formulation including a buffer and a tonicifying agent, free of a surfactant. In one embodiment, the formulation does not contain a preservative. In a preferred

embodiment, the hGH-XTEN of the formulation is the sequence of SEQ ID NO: 1. In some embodiments, the appearance of these solutions is a clear, pale yellow to pale reddish brown liquid, free from visible particles.

Suitable pH ranges, adjusted with buffer, for the aqueous hGH formulation are from about 4 to about 6, more preferably about 5 to about 6, most advantageously about 5.5.

Preferably, a buffer concentration range is chosen to minimize deamidation, aggregation, and/or precipitation of hGH-XTEN.

The buffer concentration in the formulations of the present invention is in the range of about 2 mM to about 50 mM, preferably about 10 mM to about 40 mM, more preferably about 15 mM to about 25 mM, most preferably about 20 mM. Preferably, the buffer is a histidine buffer. In yet another preferred embodiment, the buffer is L-histidine used in the formulation at a concentration of about 20 mM. Biological buffers with similar effective pH ranges, for example citrate, maleate, and bis-TRIS, may be substituted for L-histidine.

The salt concentration is adjusted to near isotonicity, depending on the other ingredients present in the formulation. For example, the concentration range of NaCl may be about 50-200 mM, depending on the other ingredients present. Of course, other tonicifying agents can also be used such as, for example, MgCl₂ or CaCl₂.

The concentration of hGH-XTEN in the formulations of the present invention is in the range of about 25 mg/ml to about 150 mg/ml. Preferably, the concentration of hGH-XTEN in the formulations is about 100 mg/ml. In yet another preferred embodiment, the concentration of hGH-XTEN in the formulations is about 150 mg/ml. The formulations herein have an osmolality between about 300 mOsm to about 600 mOsm, preferably between about 400 mOsm and about 500 mOsm.

Mannitol may optionally be included in the aqueous hGH formulations of the present invention. The preferred concentration of mannitol is about 5 mg/ml to about 50 mg/ml. As an alternative to mannitol, other sugars or sugar alcohols are used, such as lactose, trehalose, stachyose, sorbitol, xylitol, ribitol, myoinositol, galactitol, and the like.

The formulation of the invention is storage stable for at least about one month when stored at about 23±3°C, and for at least about 6 months to about 18 months when stored at about 5±3 °C. In a preferred embodiment, the formulation is storage stable for at least about 6 months to about 3 years when stored at about 5±3 °C. The oxidation of the fusion protein in the formulations herein is reduced as compared to the oxidation of the corresponding fusion protein formulated with a surfactant. The deamidation of the fusion protein in the

formulations is reduced as compared to the deamidation of the corresponding fusion protein formulated at a higher pH. Preferably, the isolated fusion protein does not show detectable oxidation or deamidation following incubation for about 8 hours at about 40 °C.

Another aspect of the invention provides a method of preparing (making) a storage stable aqueous liquid formulation in which a fusion protein comprising an extended recombinant polypeptide (XTEN) fused to a growth hormone (GH) sequence, wherein the GH sequence is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO:43, is admixed with a buffer in the absence of a surfactant, and adjusting the pH of the mixture to obtain a pH of about 4 to about 6.

A preferred embodiment of the present invention is prepared by dissolving about 25 mg/ml to about 150 mg/ml of hGH-XTEN in 950 ml sterile water for injection USP, 9.00 g NaCl (low endotoxin; Merck Chemicals #116224) USP (at 0.154M final volume), 3.103g L- Histidine (Avantor #2080) USP (at 0.02M final volume), adjusted to pH 5.5 +/- 0.1 at 18 +/- 1 C with 5M HC1. The final volume is adjusted to 1 L with sterile water for injection USP. The conductivity of the formulation solution at 25 +/- 1 C is about 10 to about 20 mS/cm.

It will be understood that the above quantities are somewhat flexible within ranges, as set forth in more detail above, and that the materials are interchangeable within the component categories. More than one buffering agent, preservative, sugar, or neutral salt may be used. Preferably, the formulation is isotonic and sterile.

In general, the formulations of the subject invention may contain other components in amounts not detracting from the preparation of stable forms and in amounts suitable for effective, safe pharmaceutical administration. For example, other pharmaceutically acceptable excipients well known to those skilled in the art may form a part of the subject compositions. These include, for example, various bulking agents, additional buffering agents, chelating agents, antioxidants, cosolvents and the like; specific examples of these could include citrate or succinate buffers, and disodium edetate. IV). GROWTH HORMONE

The present invention concerns an improved therapeutic aqueous liquid formulation comprising hGH-XTEN fusion protein. In addition, in one aspect, the present invention concerns a method of treating human growth hormone deficiency (GHD) in patients with a hGH-XTEN fusion protein formulation.

The invention contemplates hGH-XTEN in the formulations comprising sequences with homology to hGH sequences, sequence fragments that are natural, such as from humans and non-natural sequence variants which retain at least a portion of the biologic activity or biological function of hGH and/or that are useful for preventing, treating, mediating, or ameliorating a hGH-related disease, deficiency, disorder or condition in pediatric patients. In addition, native sequences homologous to human GH may be found by standard homology searching techniques, such as NCBI BLAST. The hGH of the subject compositions, together with their corresponding nucleic acid and amino acid sequences, are well known in the art, described in in, for example, W013/184216, WO/2014/164568, U.S. Patent Nos. 8,492,530 and 8,703,717, and descriptions and sequences are available in public databases such as Chemical Abstracts Services Databases (e.g., the CAS Registry), GenBank, The Universal Protein Resource (UniProt) and subscription provided databases such as GenSeq (e.g., Derwent).

Effects of hGH on the tissues of the body can generally be described as anabolic. Like most other protein hormones, native hGH acts by interacting with a specific plasma membrane receptor, referred to as growth hormone receptor. hGH acts on the liver and other tissues to stimulate production of IGF-I, which is responsible for the growth promoting effects of hGH and also reflects the amount produced. IGF-I, in turn, has stimulatory effects on osteoblast and chondrocyte activity to promote bone growth. In one embodiment, the invention provides a hGH-XTEN formulation that exhibits at least one of the properties of native hGH hereinabove described herein.

In one embodiment, the hGH incorporated into the subject hGH-XTEN formulation is a recombinant polypeptide with a sequence corresponding to a protein found in nature. In another embodiment, the hGH is a sequence variant, fragment, homolog, or a mimetics of a natural sequence that retains at least a portion of the biological activity of the corresponding native GH. In one other embodiment, the hGH is recombinant human GH comprising the following amino acid sequence:

FPTIPLSRLFDNAMLRAHRLHQLAFDTYQEFEEAYIPKEQKYSFLQNPQTSLCFSESIP TPS REETQQKSNLELLRISLLLIQSWLEPVQFLRSVFANSLVYGASDSNVYDLLKDL EEGIQTLMGRLEDGSPRTGQIFKQTYSKFDTNSHNDDALLKNYGLLYCFRKDMDKV ETFLRIVQCRSVEGSCGF (SEQ ID NO:43). Any human GH sequences or homologous derivatives constructed by shuffling individual mutations between families that retain at least a portion of the biological activity of the native GH may be useful for the fusion proteins of this invention. Human GH that can be incorporated into a hGH-XTEN fusion protein can include a protein that exhibits at least about 80% sequence identity, or alternatively 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:43.

V). HUMAN GROWTH HORMONE-XTEN FUSION PROTEIN COMPOSITIONS

In one aspect, the hGH fusion proteins suitable for use in the present invention comprise a human growth hormone polypeptide and one or more XTEN sequences as described herein, and as disclosed in, for example, W013/184216, WO/2014/164568 and U.S. Patent Nos. 8,492,530 and 8,703,717, each of which is incorporated herein by reference in its entirety.

In one other aspect, the hGH-XTEN fusion proteins are isolated monomelic fusion proteins of hGH comprising the full-length sequence or sequence variants of hGH covalently linked to one or more extended recombinant polypeptides ("XTEN" or "XTENs"). In one embodiment, the hGH-XTEN fusion protein comprises an amino acid sequence shown in FIG. 1 (SEQ ID NO: l), or pharmacologically active variants thereof. In another

embodiment, the hGH-XTEN fusion protein comprises an amino acid sequence selected from Table 1. In another embodiment, the hGH-XTEN fusion protein sequence exhibits at least about 80% sequence identity, or alternatively 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to a XTEN selected from Table 1.

For example, the hGH-XTEN fusion protein VRS-317 (SEQ ID NO: 1), is composed of recombinant human growth hormone (rhGH) and two recombinant polypeptides, referred to as XTEN as described in Schellenberger et al. (2009). Nat Biotechnol 27, 1186-90, Schellenberger et al. WO10/144502A2, and WO10/091122, each of which are incorporated herein by reference in their entirety. The XTEN domain, two unstructured hydrophilic chains of amino acids, provides half-life extension for rhGH. The molecular weight of VRS-317 is 118.9 kDa, with rhGH contributing 22.1 kDa and the remaining mass contributed by the XTEN construct. The mass ratio of rhGH to VRS-317 is therefore 1 :5.37. Further characterization of the exemplary hGH-XTEN fusion proteins provided in Table 1 can be found in WO10/144502A2, which is incorporated herein by reference in its entirety.

Table 1 - Exemplary hGH-XTEN fusion proteins

hGH- SEQ SEQ XTEN Amino Acid Sequence ID DNA Nucleotide Sequence ID Name* NO: NO:

AE912- AEPAGSPTSTEEGTPGS 1 ATGGCTGAACCTGCTGGCTCTCCAACCTCCA 7 hGH- GTASSSPGSSTPSGATG CTGAGGAAGGTACCCCGGGTAGCGGTACTG

AE144 SPGASPGTSSTGSPGSP CTTCTTCCTCTCCAGGTAGCTCTACCCCTTC

AGSPTSTEEGTSESATP TGGTGCAACCGGCTCTCCAGGTGCTTCTCCG

ESGPGTSTEPSEGSAPG GGCACCAGCTCTACCGGTTCTCCAGGTAGC

SPAGSPTSTEEGTSTEPS CCGGCTGGCTCTCCTACCTCTACTGAGGAA

EGSAPGTSTEPSEGSAP GGTACTTCTGAAAGCGCTACTCCTGAGTCTG

GTSESATPESGPGSEPA GTCCAGGTACCTCTACTGAACCGTCCGAAG

TSGSETPGSEPATSGSET GTAGCGCTCCAGGTAGCCCAGCAGGCTCTC

PGSPAGSPTSTEEGTSES CGACTTCCACTGAGGAAGGTACTTCTACTG

ATPESGPGTSTEPSEGS AACCTTCCGAAGGCAGCGCACCAGGTACCT

APGTSTEPSEGSAPGSP CTACTGAACCTTCTGAGGGCAGCGCTCCAG

AGSPTSTEEGTSTEPSE GTACTTCTGAAAGCGCTACCCCGGAATCTG

GSAPGTSTEPSEGSAPG GCCCAGGTAGCGAACCGGCTACTTCTGGTT

TSESATPESGPGTSTEPS CTGAAACCCCAGGTAGCGAACCGGCTACCT

EGSAPGTSESATPESGP CCGGTTCTGAAACTCCAGGTAGCCCGGCAG

GSEPATSGSETPGTSTEP GCTCTCCGACCTCTACTGAGGAAGGTACTTC

SEGSAPGTSTEPSEGSA TGAAAGCGCAACCCCGGAGTCCGGCCCAGG

PGTSESATPESGPGTSES TACCTCTACCGAACCGTCTGAGGGCAGCGC

ATPESGPGSPAGSPTST ACCAGGTACTTCTACCGAACCGTCCGAGGG

EEGTSESATPESGPGSEP TAGCGCACCAGGTAGCCCAGCAGGTTCTCC

ATSGSETPGTSESATPES TACCTCCACCGAGGAAGGTACTTCTACCGA

GPGTSTEPSEGSAPGTS ACCGTCCGAGGGTAGCGCACCAGGTACCTC

TEPSEGSAPGTSTEPSEG TACTGAACCTTCTGAGGGCAGCGCTCCAGG

SAPGTSTEPSEGSAPGT TACTTCTGAAAGCGCTACCCCGGAGTCCGG

STEPSEGSAPGTSTEPSE TCCAGGTACTTCTACTGAACCGTCCGAAGG

GSAPGSPAGSPTSTEEG TAGCGCACCAGGTACTTCTGAAAGCGCAAC

TSTEPSEGSAPGTSESAT CCCTGAATCCGGTCCAGGTAGCGAACCGGC

PESGPGSEPATSGSETP TACTTCTGGCTCTGAGACTCCAGGTACTTCT

GTSESATPESGPGSEPA ACCGAACCGTCCGAAGGTAGCGCACCAGGT

TSGSETPGTSESATPESG ACTTCTACTGAACCGTCTGAAGGTAGCGCA

PGTSTEPSEGSAPGTSES CCAGGTACTTCTGAAAGCGCAACCCCGGAA

ATPESGPGSPAGSPTST TCCGGCCCAGGTACCTCTGAAAGCGCAACC

EEGSPAGSPTSTEEGSP CCGGAGTCCGGCCCAGGTAGCCCTGCTGGC

AGSPTSTEEGTSESATP TCTCCAACCTCCACCGAAGAAGGTACCTCT

ESGPGTSTEPSEGSAPG GAAAGCGCAACCCCTGAATCCGGCCCAGGT

TSESATPESGPGSEPATS AGCGAACCGGCAACCTCCGGTTCTGAAACC

GSETPGTSESATPESGP CCAGGTACCTCTGAAAGCGCTACTCCGGAG

GSEPATSGSETPGTSES TCTGGCCCAGGTACCTCTACTGAACCGTCTG

ATPESGPGTSTEPSEGS AGGGTAGCGCTCCAGGTACTTCTACTGAAC

APGSPAGSPTSTEEGTS CGTCCGAAGGTAGCGCACCAGGTACTTCTA

ESATPESGPGSEPATSG CCGAACCGTCCGAAGGCAGCGCTCCAGGTA

SETPGTSESATPESGPGS CCTCTACTGAACCTTCCGAGGGCAGCGCTC

PAGSPTSTEEGSPAGSP CAGGTACCTCTACCGAACCTTCTGAAGGTA

TSTEEGTSTEPSEGSAP GCGCACCAGGTACTTCTACCGAACCGTCCG

GTSESATPESGPGTSES AGGGTAGCGCACCAGGTAGCCCAGCAGGTT

ATPESGPGTSESATPES CTCCTACCTCCACCGAGGAAGGTACTTCTAC hGH- SEQ SEQ XTEN Amino Acid Sequence ID DNA Nucleotide Sequence ID Name* NO: NO:

GPGSEPATSGSETPGSE CGAACCGTCCGAGGGTAGCGCACCAGGTAC

PATSGSETPGSPAGSPTS CTCTGAAAGCGCAACTCCTGAGTCTGGCCC

TEEGTSTEPSEGSAPGT AGGTAGCGAACCTGCTACCTCCGGCTCTGA

STEPSEGSAPGSEPATS GACTCCAGGTACCTCTGAAAGCGCAACCCC

GSETPGTSESATPESGP GGAATCTGGTCCAGGTAGCGAACCTGCAAC

GTSTEPSEGSAPGFPTIP CTCTGGCTCTGAAACCCCAGGTACCTCTGA

LSRLFDNAMLRAHRLH AAGCGCTACTCCTGAATCTGGCCCAGGTAC

QLAFDTYQEFEEAYIPK TTCTACTGAACCGTCCGAGGGCAGCGCACC

EQKYSFLQNPQTSLCFS AGGTACTTCTGAAAGCGCTACTCCTGAGTC

ESIPTP SNREETQQKSNL CGGCCCAGGTAGCCCGGCTGGCTCTCCGAC

ELLPJSLLLIQSWLEPVQ TTCCACCGAGGAAGGTAGCCCGGCTGGCTC

FLRS VF ANSL VYG ASD S TCCAACTTCTACTGAAGAAGGTAGCCCGGC

NVYDLLKDLEEGIQTL AGGCTCTCCGACCTCTACTGAGGAAGGTAC

MGRLEDGSPRTGQIFK TTCTGAAAGCGCAACCCCGGAGTCCGGCCC

QTYSKFDTNSHNDDAL AGGTACCTCTACCGAACCGTCTGAGGGCAG

LKNYGLLYCFRKDMD CGCACCAGGTACCTCTGAAAGCGCAACTCC

KVETFLRIVQCRSVEGS TGAGTCTGGCCCAGGTAGCGAACCTGCTAC

CGFGGTSESATPESGPG CTCCGGCTCTGAGACTCCAGGTACCTCTGA

TSTEPSEGSAPGTSTEPS AAGCGCAACCCCGGAATCTGGTCCAGGTAG

EGSAPGTSESATPESGP CGAACCTGCAACCTCTGGCTCTGAAACCCC

GTSTEPSEGSAPGTSTEP AGGTACCTCTGAAAGCGCTACTCCTGAATC

SEGSAPGTSESATPESG TGGCCCAGGTACTTCTACTGAACCGTCCGA

PGTSTEPSEGSAPGTSTE GGGCAGCGCACCAGGTAGCCCTGCTGGCTC

PSEGSAPGTSTEPSEGS TCCAACCTCCACCGAAGAAGGTACCTCTGA

APGSPAGSPTSTEEGTS AAGCGCAACCCCTGAATCCGGCCCAGGTAG

TEPSEGSAPG CGAACCGGCAACCTCCGGTTCTGAAACCCC

AGGTACTTCTGAAAGCGCTACTCCTGAGTC

CGGCCCAGGTAGCCCGGCTGGCTCTCCGAC

TTCCACCGAGGAAGGTAGCCCGGCTGGCTC

TCCAACTTCTACTGAAGAAGGTACTTCTACC

GAACCTTCCGAGGGCAGCGCACCAGGTACT

TCTGAAAGCGCTACCCCTGAGTCCGGCCCA

GGTACTTCTGAAAGCGCTACTCCTGAATCC

GGTCCAGGTACTTCTGAAAGCGCTACCCCG

GAATCTGGCCCAGGTAGCGAACCGGCTACT

TCTGGTTCTGAAACCCCAGGTAGCGAACCG

GCTACCTCCGGTTCTGAAACTCCAGGTAGC

CCAGCAGGCTCTCCGACTTCCACTGAGGAA

GGTACTTCTACTGAACCTTCCGAAGGCAGC

GCACCAGGTACCTCTACTGAACCTTCTGAG

GGCAGCGCTCCAGGTAGCGAACCTGCAACC

TCTGGCTCTGAAACCCCAGGTACCTCTGAA

AGCGCTACTCCTGAATCTGGCCCAGGTACTT

CTACTGAACCGTCCGAGGGCAGCGCACCAG

GTTTTCCGACTATTCCGCTGTCTCGTCTGTTT

GATAATGCTATGCTGCGTGCGCACCGTCTG

CACCAGCTGGCCTTTGATACTTACCAGGAA

TTTGAAGAAGCcTACATTCCTAAAGAGCAG

AAGTACTCTTTCCTGCAAAACCCACAGACTT

CTCTCTGCTTCAGCGAATCTATTCCGACGCC

TTCCAATCGCGAGGAAACTCAGCAAAAGTC

CAATCTGGAACTACTCCGCATTTCTCTGCTT

CTGATTCAGAGCTGGCTAGAACCAGTGCAA

TTTCTGCGTTCCGTCTTCGCCAATAGCCTAG

TTTATGGCGCATCCGACAGCAACGTATACG

ATCTCCTGAAAGATCTCGAGGAAGGCATTC hGH- SEQ SEQ XTEN Amino Acid Sequence ID DNA Nucleotide Sequence ID Name* NO: NO:

AGACCCTGATGGGTCGTCTCGAGGATGGCT

CTCCGCGTACTGGTCAGATCTTCAAGCAGA

CTTACTCTAAATTTGATACTAACAGCCACAA

TGACGATGCGCTTCTAAAAAACTATGGTCT

GCTGTATTGTTTTCGTAAAGATATGGACAA

AGTTGAAACCTTCCTGCGTATTGTTCAGTGT

CGTTCCGTTGAGGGCAGCTGTGGTTTCTAAG

GTGGTAGCGAACCGGCAACTTCCGGCTCTG

AAACCCCAGGTACTTCTGAAAGCGCTACTC

CTGAGTCTGGCCCAGGTAGCGAACCTGCTA

CCTCTGGCTCTGAAACCCCAGGTAGCCCGG

CAGGCTCTCCGACTTCCACCGAGGAAGGTA

CCTCTACTGAACCTTCTGAGGGTAGCGCTCC

AGGTAGCGAACCGGCAACCTCTGGCTCTGA

AACCCCAGGTAGCGAACCTGCTACCTCCGG

CTCTGAAACTCCAGGTAGCGAACCGGCTAC

TTCCGGTTCTGAAACTCCAGGTACCTCTACC

GAACCTTCCGAAGGCAGCGCACCAGGTACT

TCTGAAAGCGCAACCCCTGAATCCGGTCCA

GGTAGCGAACCGGCTACTTCTGGCTCTGAG

ACTCCAGGTACTTCTACCGAACCGTCCGAA

GGTAGCGCACCA

AM864- GGSPGTSTEPSEGSAPG 2 ggtGGGTCTCCAGGTACTTCTACTGAACCGTC 8 hGH SEPATSGSETPGSPAGSP TGAAGGCAGCGCACCAGGTAGCGAACCGGC

TSTEEGSTSSTAESPGPG TACTTCCGGTTCTGAAACCCCAGGTAGCCC

TSTPESGSASPGSTSESP AGCAGGTTCTCCAACTTCTACTGAAGAAGG

SGTAPGSTSESPSGTAP TTCTACCAGCTCTACCGCAGAATCTCCTGGT

GTSTPESGSASPGTSTPE CCAGGTACCTCTACTCCGGAAAGCGGCTCT

SGSASPGSEPATSGSETP GCATCTCCAGGTTCTACTAGCGAATCTCCTT

GTSESATPESGPGSPAG CTGGCACTGCACCAGGTTCTACTAGCGAAT

SPTSTEEGTSTEPSEGSA CCCCGTCTGGTACTGCTCCAGGTACTTCTAC

PGTSESATPESGPGTSTE TCCTGAAAGCGGTTCCGCTTCTCCAGGTACC

PSEGSAPGTSTEPSEGS TCTACTCCGGAAAGCGGTTCTGCATCTCCAG

APGSPAGSPTSTEEGTS GTAGCGAACCGGCAACCTCCGGCTCTGAAA

TEPSEGSAPGTSTEPSEG CCCCAGGTACCTCTGAAAGCGCTACTCCTG

SAPGTSESATPESGPGT AATCCGGCCCAGGTAGCCCGGCAGGTTCTC

SESATPESGPGTSTEPSE CGACTTCCACTGAGGAAGGTACCTCTACTG

GSAPGTSTEPSEGSAPG AACCTTCTGAGGGCAGCGCTCCAGGTACTT

TSESATPESGPGTSTEPS CTGAAAGCGCTACCCCGGAGTCCGGTCCAG

EGSAPGSEPATSGSETP GTACTTCTACTGAACCGTCCGAAGGTAGCG

GSPAGSPTSTEEGS STPS CACCAGGTACTTCTACCGAACCGTCCGAGG

GATGSPGTPGSGTASSS GTAGCGCACCAGGTAGCCCAGCAGGTTCTC

PGSSTPSGATGSPGTST CTACCTCCACCGAGGAAGGTACTTCTACCG

EPSEGSAPGTSTEPSEGS AACCGTCCGAGGGTAGCGCACCAGGTACTT

APGSEPATSGSETPGSP CTACCGAACCTTCCGAGGGCAGCGCACCAG

AGSPTSTEEGSPAGSPT GTACTTCTGAAAGCGCTACCCCTGAGTCCG

STEEGTSTEPSEGSAPG GCCCAGGTACTTCTGAAAGCGCTACTCCTG

ASASGAPSTGGTSESAT AATCCGGTCCAGGTACCTCTACTGAACCTTC

PESGPGSPAGSPTSTEE CGAAGGCAGCGCTCCAGGTACCTCTACCGA

GSPAGSPTSTEEGSTSST ACCGTCCGAGGGCAGCGCACCAGGTACTTC

AESPGPGSTSESPSGTAP TGAAAGCGCAACCCCTGAATCCGGTCCAGG

GTSPSGESSTAPGTPGS TACTTCTACTGAACCTTCCGAAGGTAGCGCT

GTASSSPGSSTPSGATG CCAGGTAGCGAACCTGCTACTTCTGGTTCTG

SPGSSPSASTGTGPGSEP AAACCCCAGGTAGCCCGGCTGGCTCTCCGA

ATSGSETPGTSESATPES CCTCCACCGAGGAAGGTAGCTCTACCCCGT

GPGSEPATSGSETPGST CTGGTGCTACTGGTTCTCCAGGTACTCCGGG hGH- SEQ SEQ XTEN Amino Acid Sequence ID DNA Nucleotide Sequence ID Name* NO: NO:

SSTAESPGPGSTSSTAES CAGCGGTACTGCTTCTTCCTCTCCAGGTAGC

PGPGTSPSGESSTAPGSE TCTACCCCTTCTGGTGCTACTGGCTCTCCAG

PATSGSETPGSEPATSG GTACCTCTACCGAACCGTCCGAGGGTAGCG

SETPGTSTEPSEGSAPGS CACCAGGTACCTCTACTGAACCGTCTGAGG

TSSTAESPGPGTSTPESG GTAGCGCTCCAGGTAGCGAACCGGCAACCT

SASPGSTSESPSGTAPGT CCGGTTCTGAAACTCCAGGTAGCCCTGCTG

STEPSEGSAPGTSTEPSE GCTCTCCGACTTCTACTGAGGAAGGTAGCC

GSAPGTSTEPSEGSAPG CGGCTGGTTCTCCGACTTCTACTGAGGAAG

SSTPSGATGSPGSSPSAS GTACTTCTACCGAACCTTCCGAAGGTAGCG

TGTGPGASPGTSSTGSP CTCCAGGTGCAAGCGCAAGCGGCGCGCCAA

GSEPATSGSETPGTSES GCACGGGAGGTACTTCTGAAAGCGCTACTC

ATPESGPGSPAGSPTST CTGAGTCCGGCCCAGGTAGCCCGGCTGGCT

EEGSSTPSGATGSPGSSP CTCCGACTTCCACCGAGGAAGGTAGCCCGG

SASTGTGPGASPGTSST CTGGCTCTCCAACTTCTACTGAAGAAGGTTC

GSPGTSESATPESGPGT TACCAGCTCTACCGCTGAATCTCCTGGCCCA

STEPSEGSAPGTSTEPSE GGTTCTACTAGCGAATCTCCGTCTGGCACCG

GSAPGFPTIPLSRLFDNA CACCAGGTACTTCCCCTAGCGGTGAATCTTC

MLRAHRLHQLAFDTYQ TACTGCACCAGGTACCCCTGGCAGCGGTAC

EFEE AYIPKEQKY SFLQ CGCTTCTTCCTCTCCAGGTAGCTCTACCCCG

NPQTSLCFSESIPTPSNR TCTGGTGCTACTGGCTCTCCAGGTTCTAGCC

EETQQKSNLELLPJSLL CGTCTGCATCTACCGGTACCGGCCCAGGTA

LIQSWLEPVQFLRSVFA GCGAACCGGCAACCTCCGGCTCTGAAACTC

NSL VYG ASD SNVYDLL CAGGTACTTCTGAAAGCGCTACTCCGGAAT

KDLEEGIQTLMGRLED CCGGCCCAGGTAGCGAACCGGCTACTTCCG

GSPRTGQIFKQTYSKFD GCTCTGAAACCCCAGGTTCCACCAGCTCTA

TNSHNDDALLKNYGLL CTGCAGAATCTCCGGGCCCAGGTTCTACTA

YCFRKDMDKVETFLRI GCTCTACTGCAGAATCTCCGGGTCCAGGTA

VQCRSVEGSCGF CTTCTCCTAGCGGCGAATCTTCTACCGCTCC

AGGTAGCGAACCGGCAACCTCTGGCTCTGA

AACTCCAGGTAGCGAACCTGCAACCTCCGG

CTCTGAAACCCCAGGTACTTCTACTGAACCT

TCTGAGGGCAGCGCACCAGGTTCTACCAGC

TCTACCGCAGAATCTCCTGGTCCAGGTACCT

CTACTCCGGAAAGCGGCTCTGCATCTCCAG

GTTCTACTAGCGAATCTCCTTCTGGCACTGC

ACCAGGTACTTCTACCGAACCGTCCGAAGG

CAGCGCTCCAGGTACCTCTACTGAACCTTCC

GAGGGCAGCGCTCCAGGTACCTCTACCGAA

CCTTCTGAAGGTAGCGCACCAGGTAGCTCT

ACTCCGTCTGGTGCAACCGGCTCCCCAGGTT

CTAGCCCGTCTGCTTCCACTGGTACTGGCCC

AGGTGCTTCCCCGGGCACCAGCTCTACTGG

TTCTCCAGGTAGCGAACCTGCTACCTCCGGT

TCTGAAACCCCAGGTACCTCTGAAAGCGCA

ACTCCGGAGTCTGGTCCAGGTAGCCCTGCA

GGTTCTCCTACCTCCACTGAGGAAGGTAGC

TCTACTCCGTCTGGTGCAACCGGCTCCCCAG

GTTCTAGCCCGTCTGCTTCCACTGGTACTGG

CCCAGGTGCTTCCCCGGGCACCAGCTCTACT

GGTTCTCCAGGTACCTCTGAAAGCGCTACTC

CGGAGTCTGGCCCAGGTACCTCTACTGAAC

CGTCTGAGGGTAGCGCTCCAGGTACTTCTA

CTGAACCGTCCGAAGGTAGCGCACCAGGTT

TTCCGACTATTCCGCTGTCTCGTCTGTTTGA

TAATGCTATGCTGCGTGCGCACCGTCTGCAC

CAGCTGGCCTTTGATACTTACCAGGAATTTG hGH- SEQ SEQ XTEN Amino Acid Sequence ID DNA Nucleotide Sequence ID Name* NO: NO:

AAGAAGCcTACATTCCTAAAGAGCAGAAGT

ACTCTTTCCTGCAAAACCCACAGACTTCTCT

CTGCTTCAGCGAATCTATTCCGACGCCTTCC

AATCGCGAGGAAACTCAGCAAAAGTCCAAT

CTGGAACTACTCCGCATTTCTCTGCTTCTGA

TTCAGAGCTGGCTAGAACCAGTGCAATTTC

TGCGTTCCGTCTTCGCCAATAGCCTAGTTTA

TGGCGCATCCGACAGCAACGTATACGATCT

CCTGAAAGATCTCGAGGAAGGCATTCAGAC

CCTGATGGGTCGTCTCGAGGATGGCTCTCC

GCGTACTGGTCAGATCTTCAAGCAGACTTA

CTCTAAATTTGATACTAACAGCCACAATGA

CGATGCGCTTCTAAAAAACTATGGTCTGCT

GTATTGTTTTCGTAAAGATATGGACAAAGTT

GAAACCTTCCTGCGTATTGTTCAGTGTCGTT

CCGTTGAGGGCAGCTGTGGTTTC

Y576- GEGSGEGSEGEGSEGSG 3 GGTGAGGGTTCTGGCGAAGGTTCCGAAGGT 9 hGH EGEGSEGSGEGEGGSE GAGGGCTCCGAAGGATCTGGCGAAGGTGAG

GSEGEGSEGSGEGEGG GGTTCCGAAGGTTCTGGCGAAGGTGAAGGC

EGSGEGEGSGEGSEGE GGTTCTGAGGGATCCGAAGGTGAAGGCTCC

GGGEGSEGEGSGEGGE GAAGGATCTGGCGAAGGTGAAGGTGGTGA

GEGSEGGSEGEGGSEG AGGTTCTGGCGAAGGTGAGGGATCTGGCGA

GEGEGSEGSGEGEGSE AGGCTCTGAAGGTGAAGGTGGTGGTGAAGG

GGSEGEGSEGGSEGEGS CTCTGAAGGTGAAGGATCTGGTGAAGGTGG

EGSGEGEGSEGSGEGE CGAAGGTGAGGGATCTGAAGGCGGCTCCGA

GSEGSGEGEGSEGSGEG AGGTGAAGGCGGATCTGAAGGCGGCGAAG

EGSEGGSEGEGGSEGSE GTGAAGGTTCCGAAGGTTCTGGTGAAGGTG

GEGSGEGSEGEGGSEGS AAGGATCTGAAGGTGGCTCCGAAGGTGAAG

EGEGGGEGSEGEGSGE GATCTGAAGGCGGTTCCGAAGGTGAGGGCT

GSEGEGGSEGSEGEGGS CTGAAGGTTCTGGCGAAGGTGAAGGCTCTG

EGSEGEGGEGSGEGEG AAGGATCTGGTGAAGGTGAAGGTTCCGAAG

SEGSGEGEGSGEGSEGE GTTCTGGTGAAGGTGAAGGTTCCGAAGGTT

GSEGSGEGEGSEGSGEG CTGGCGAAGGTGAAGGTTCTGAAGGTGGCT

EGGSEGSEGEGSGEGSE CTGAAGGTGAAGGCGGCTCTGAAGGATCCG

GEGSEGSGEGEGSEGSG AAGGTGAAGGTTCTGGTGAAGGCTCTGAAG

EGEGGSEGSEGEGGSE GTGAAGGCGGCTCTGAGGGTTCCGAAGGTG

GSEGEGGSEGSEGEGG AAGGCGGAGGCGAAGGTTCTGAAGGTGAG

EGSGEGEGSEGSGEGE GGATCTGGTGAAGGTTCTGAAGGTGAAGGC

GSGEGSEGEGSEGSGEG GGTTCTGAAGGTTCCGAAGGTGAAGGTGGC

EGSEGSGEGEGGSEGSE TCTGAGGGATCCGAAGGTGAAGGTGGCGAA

GEGSEGSGEGEGGEGS GGATCTGGTGAAGGTGAAGGTTCTGAAGGT

GEGEGSGEGSEGEGGG TCTGGCGAAGGTGAGGGTTCTGGCGAAGGT

EGSEGEGSEGSGEGEGS TCCGAAGGTGAGGGCTCCGAAGGATCTGGC

EGSGEGEGSEGGSEGE GAAGGTGAGGGTTCCGAAGGTTCTGGCGAA

GGSEGSEGEGSEGGSEG GGTGAAGGCGGTTCTGAGGGATCCGAAGGT

EGSEGGSEGEGSEGSGE GAGGGTTCTGGCGAAGGTTCCGAAGGTGAG

GEGSEGSGEGEGSGEGS GGCTCCGAAGGATCTGGCGAAGGTGAGGGT

EGEGGSEGGEGEGSEG TCCGAAGGTTCTGGCGAAGGTGAAGGCGGT

GSEGEGSEGGSEGEGG TCTGAGGGATCCGAAGGTGAAGGCGGTTCT

EGSGEGEGGGEGSEGE GAAGGTTCCGAAGGTGAAGGTGGCTCTGAG

GSEGSGEGEGSGEGSEG GGATCCGAAGGTGAAGGTGGCGAAGGATCT

FPTIPLSRLFDNAMLRA GGTGAAGGTGAAGGTTCTGAAGGTTCTGGC

HRLHQLAFDTYQEFEE GAAGGTGAGGGTTCTGGCGAAGGTTCCGAA

AYIPKEQKYSFLQNPQT GGTGAGGGCTCCGAAGGATCTGGCGAAGGT

SLCFSESIPTPSNREETQ GAGGGTTCCGAAGGTTCTGGCGAAGGTGAA

QKSNLELLRISLLLIQS GGCGGTTCTGAGGGATCCGAAGGTGAAGGC hGH- SEQ SEQ XTEN Amino Acid Sequence ID DNA Nucleotide Sequence ID Name* NO: NO:

WLEPVQFLRSVFANSL TCCGAAGGATCTGGCGAAGGTGAAGGTGGT

VYGASDSNVYDLLKDL GAAGGTTCTGGCGAAGGTGAGGGATCTGGC

EEGIQTLMGRLEDGSPR GAAGGCTCTGAAGGTGAAGGTGGTGGTGAA

TGQIFKQTYSKFDTNSH GGCTCTGAAGGTGAAGGTTCCGAAGGTTCT

NDDALLKNYGLLYCFR GGTGAAGGTGAAGGTTCCGAAGGTTCTGGC

KDMDKVETFLRIVQCR GAAGGTGAAGGTTCTGAAGGTGGCTCTGAA

SVEGSCGF GGTGAAGGCGGCTCTGAAGGATCCGAAGGT

GAAGGATCTGAAGGTGGCTCCGAAGGTGAA

GGATCTGAAGGCGGTTCCGAAGGTGAGGGC

TCTGAAGGTTCTGGCGAAGGTGAAGGCTCT

GAAGGATCTGGTGAAGGTGAAGGATCTGGC

GAAGGCTCCGAAGGTGAAGGCGGTTCTGAA

GGTGGCGAAGGTGAAGGATCTGAAGGTGGT

TCCGAAGGTGAGGGATCTGAAGGTGGCTCT

GAAGGTGAAGGTGGCGAAGGTTCTGGCGAA

GGTGAAGGTGGAGGCGAAGGTTCTGAAGGT

GAAGGTTCCGAAGGTTCTGGTGAAGGTGAG

GGATCTGGCGAAGGTTCTGAAGGTTTTCCG

ACTATTCCGCTGTCTCGTCTGTTTGATAACG

CTATGCTGCGTGCGCACCGTCTGCACCAGCT

GGCGTTCGACACTTACCAGGAATTTGAAGA

AGCGTACATTCCGAAGGAACAGAAGTACTC

TTTCCTGCAAAACCCGCAGACCTCCCTGTGC

TTCAGCGAATCTATTCCGACTCCGTCCAATC

GTGAAGAAACTCAGCAAAAGTCCAATCTGG

AGCTGCTGCGCATCTCTCTGCTGCTGATTCA

GAGCTGGCTGGAGCCTGTTCAGTTTCTGCGT

TCCGTCTTCGCCAACAGCCTGGTTTATGGTG

CTTCCGACAGCAACGTATACGATCTGCTGA

AAGATCTGGAAGAAGGCATTCAGACCCTGA

TGGGTCGTCTGGAAGATGGTTCTCCGCGTA

CTGGTCAGATCTTCAAACAAACTTACTCCA

AATTTGATACTAACAGCCATAACGACGATG

CTCTGCTGAAAAACTATGGTCTGCTGTATTG

CTTCCGCAAGGATATGGACAAAGTTGAAAC

CTTCCTGCGTATTGTGCAGTGTCGTTCCGTT

GAGGGCAGCTGTGGTTTC

AE912- AEPAGSPTSTEEGTPGS 4 ATGGCTGAACCTGCTGGCTCTCCAACCTCCA 10 hGH GTASSSPGSSTPSGATG CTGAGGAAGGTACCCCGGGTAGCGGTACTG

SPGASPGTSSTGSPGSP CTTCTTCCTCTCCAGGTAGCTCTACCCCTTC

AGSPTSTEEGTSESATP TGGTGCAACCGGCTCTCCAGGTGCTTCTCCG

ESGPGTSTEPSEGSAPG GGCACCAGCTCTACCGGTTCTCCAGGTAGC

SPAGSPTSTEEGTSTEPS CCGGCTGGCTCTCCTACCTCTACTGAGGAA

EGSAPGTSTEPSEGSAP GGTACTTCTGAAAGCGCTACTCCTGAGTCTG

GTSESATPESGPGSEPA GTCCAGGTACCTCTACTGAACCGTCCGAAG

TSGSETPGSEPATSGSET GTAGCGCTCCAGGTAGCCCAGCAGGCTCTC

PGSPAGSPTSTEEGTSES CGACTTCCACTGAGGAAGGTACTTCTACTG

ATPESGPGTSTEPSEGS AACCTTCCGAAGGCAGCGCACCAGGTACCT

APGTSTEPSEGSAPGSP CTACTGAACCTTCTGAGGGCAGCGCTCCAG

AGSPTSTEEGTSTEPSE GTACTTCTGAAAGCGCTACCCCGGAATCTG

GSAPGTSTEPSEGSAPG GCCCAGGTAGCGAACCGGCTACTTCTGGTT

TSESATPESGPGTSTEPS CTGAAACCCCAGGTAGCGAACCGGCTACCT

EGSAPGTSESATPESGP CCGGTTCTGAAACTCCAGGTAGCCCGGCAG

GSEPATSGSETPGTSTEP GCTCTCCGACCTCTACTGAGGAAGGTACTTC

SEGSAPGTSTEPSEGSA TGAAAGCGCAACCCCGGAGTCCGGCCCAGG

PGTSESATPESGPGTSES TACCTCTACCGAACCGTCTGAGGGCAGCGC hGH- SEQ SEQ XTEN Amino Acid Sequence ID DNA Nucleotide Sequence ID Name* NO: NO:

ATPESGPGSPAGSPTST ACCAGGTACTTCTACCGAACCGTCCGAGGG

EEGTSESATPESGPGSEP TAGCGCACCAGGTAGCCCAGCAGGTTCTCC

ATSGSETPGTSESATPES TACCTCCACCGAGGAAGGTACTTCTACCGA

GPGTSTEPSEGSAPGTS ACCGTCCGAGGGTAGCGCACCAGGTACCTC

TEPSEGSAPGTSTEPSEG TACTGAACCTTCTGAGGGCAGCGCTCCAGG

SAPGTSTEPSEGSAPGT TACTTCTGAAAGCGCTACCCCGGAGTCCGG

STEPSEGSAPGTSTEPSE TCCAGGTACTTCTACTGAACCGTCCGAAGG

GSAPGSPAGSPTSTEEG TAGCGCACCAGGTACTTCTGAAAGCGCAAC

TSTEPSEGSAPGTSESAT CCCTGAATCCGGTCCAGGTAGCGAACCGGC

PESGPGSEPATSGSETP TACTTCTGGCTCTGAGACTCCAGGTACTTCT

GTSESATPESGPGSEPA ACCGAACCGTCCGAAGGTAGCGCACCAGGT

TSGSETPGTSESATPESG ACTTCTACTGAACCGTCTGAAGGTAGCGCA

PGTSTEPSEGSAPGTSES CCAGGTACTTCTGAAAGCGCAACCCCGGAA

ATPESGPGSPAGSPTST TCCGGCCCAGGTACCTCTGAAAGCGCAACC

EEGSPAGSPTSTEEGSP CCGGAGTCCGGCCCAGGTAGCCCTGCTGGC

AGSPTSTEEGTSESATP TCTCCAACCTCCACCGAAGAAGGTACCTCT

ESGPGTSTEPSEGSAPG GAAAGCGCAACCCCTGAATCCGGCCCAGGT

TSESATPESGPGSEPATS AGCGAACCGGCAACCTCCGGTTCTGAAACC

GSETPGTSESATPESGP CCAGGTACCTCTGAAAGCGCTACTCCGGAG

GSEPATSGSETPGTSES TCTGGCCCAGGTACCTCTACTGAACCGTCTG

ATPESGPGTSTEPSEGS AGGGTAGCGCTCCAGGTACTTCTACTGAAC

APGSPAGSPTSTEEGTS CGTCCGAAGGTAGCGCACCAGGTACTTCTA

ESATPESGPGSEPATSG CCGAACCGTCCGAAGGCAGCGCTCCAGGTA

SETPGTSESATPESGPGS CCTCTACTGAACCTTCCGAGGGCAGCGCTC

PAGSPTSTEEGSPAGSP CAGGTACCTCTACCGAACCTTCTGAAGGTA

TSTEEGTSTEPSEGSAP GCGCACCAGGTACTTCTACCGAACCGTCCG

GTSESATPESGPGTSES AGGGTAGCGCACCAGGTAGCCCAGCAGGTT

ATPESGPGTSESATPES CTCCTACCTCCACCGAGGAAGGTACTTCTAC

GPGSEPATSGSETPGSE CGAACCGTCCGAGGGTAGCGCACCAGGTAC

PATSGSETPGSPAGSPTS CTCTGAAAGCGCAACTCCTGAGTCTGGCCC

TEEGTSTEPSEGSAPGT AGGTAGCGAACCTGCTACCTCCGGCTCTGA

STEPSEGSAPGSEPATS GACTCCAGGTACCTCTGAAAGCGCAACCCC

GSETPGTSESATPESGP GGAATCTGGTCCAGGTAGCGAACCTGCAAC

GTSTEPSEGSAPGFPTIP CTCTGGCTCTGAAACCCCAGGTACCTCTGA

LSRLFDNAMLRAHRLH AAGCGCTACTCCTGAATCTGGCCCAGGTAC

QLAFDTYQEFEEAYIPK TTCTACTGAACCGTCCGAGGGCAGCGCACC

EQKYSFLQNPQTSLCFS AGGTACTTCTGAAAGCGCTACTCCTGAGTC

ESIPTP SNREETQQKSNL CGGCCCAGGTAGCCCGGCTGGCTCTCCGAC

ELLPJSLLLIQSWLEPVQ TTCCACCGAGGAAGGTAGCCCGGCTGGCTC

FLRS VF ANSL VYG ASD S TCCAACTTCTACTGAAGAAGGTAGCCCGGC

NVYDLLKDLEEGIQTL AGGCTCTCCGACCTCTACTGAGGAAGGTAC

MGRLEDGSPRTGQIFK TTCTGAAAGCGCAACCCCGGAGTCCGGCCC

QTYSKFDTNSHNDDAL AGGTACCTCTACCGAACCGTCTGAGGGCAG

LKNYGLLYCFRKDMD CGCACCAGGTACCTCTGAAAGCGCAACTCC

KVETFLRIVQCRSVEGS TGAGTCTGGCCCAGGTAGCGAACCTGCTAC

CGF CTCCGGCTCTGAGACTCCAGGTACCTCTGA

AAGCGCAACCCCGGAATCTGGTCCAGGTAG

CGAACCTGCAACCTCTGGCTCTGAAACCCC

AGGTACCTCTGAAAGCGCTACTCCTGAATC

TGGCCCAGGTACTTCTACTGAACCGTCCGA

GGGCAGCGCACCAGGTAGCCCTGCTGGCTC

TCCAACCTCCACCGAAGAAGGTACCTCTGA

AAGCGCAACCCCTGAATCCGGCCCAGGTAG

CGAACCGGCAACCTCCGGTTCTGAAACCCC

AGGTACTTCTGAAAGCGCTACTCCTGAGTC

CGGCCCAGGTAGCCCGGCTGGCTCTCCGAC hGH- SEQ SEQ XTEN Amino Acid Sequence ID DNA Nucleotide Sequence ID Name* NO: NO:

TTCCACCGAGGAAGGTAGCCCGGCTGGCTC

TCCAACTTCTACTGAAGAAGGTACTTCTACC

GAACCTTCCGAGGGCAGCGCACCAGGTACT

TCTGAAAGCGCTACCCCTGAGTCCGGCCCA

GGTACTTCTGAAAGCGCTACTCCTGAATCC

GGTCCAGGTACTTCTGAAAGCGCTACCCCG

GAATCTGGCCCAGGTAGCGAACCGGCTACT

TCTGGTTCTGAAACCCCAGGTAGCGAACCG

GCTACCTCCGGTTCTGAAACTCCAGGTAGC

CCAGCAGGCTCTCCGACTTCCACTGAGGAA

GGTACTTCTACTGAACCTTCCGAAGGCAGC

GCACCAGGTACCTCTACTGAACCTTCTGAG

GGCAGCGCTCCAGGTAGCGAACCTGCAACC

TCTGGCTCTGAAACCCCAGGTACCTCTGAA

AGCGCTACTCCTGAATCTGGCCCAGGTACTT

CTACTGAACCGTCCGAGGGCAGCGCACCAG

GTTTTCCGACTATTCCGCTGTCTCGTCTGTTT

GATAATGCTATGCTGCGTGCGCACCGTCTG

CACCAGCTGGCCTTTGATACTTACCAGGAA

TTTGAAGAAGCcTACATTCCTAAAGAGCAG

AAGTACTCTTTCCTGCAAAACCCACAGACTT

CTCTCTGCTTCAGCGAATCTATTCCGACGCC

TTCCAATCGCGAGGAAACTCAGCAAAAGTC

CAATCTGGAACTACTCCGCATTTCTCTGCTT

CTGATTCAGAGCTGGCTAGAACCAGTGCAA

TTTCTGCGTTCCGTCTTCGCCAATAGCCTAG

TTTATGGCGCATCCGACAGCAACGTATACG

ATCTCCTGAAAGATCTCGAGGAAGGCATTC

AGACCCTGATGGGTCGTCTCGAGGATGGCT

CTCCGCGTACTGGTCAGATCTTCAAGCAGA

CTTACTCTAAATTTGATACTAACAGCCACAA

TGACGATGCGCTTCTAAAAAACTATGGTCT

GCTGTATTGTTTTCGTAAAGATATGGACAA

AGTTGAAACCTTCCTGCGTATTGTTCAGTGT

CGTTCCGTTGAGGGCAGCTGTGGTTTCTAA

AE912- AEPAGSPTSTEEGTPGS 5 ATGGCTGAACCTGCTGGCTCTCCAACCTCCA 11 hGH- GTASSSPGSSTPSGATG CTGAGGAAGGTACCCCGGGTAGCGGTACTG

AE288 SPGASPGTSSTGSPGSP CTTCTTCCTCTCCAGGTAGCTCTACCCCTTC

AGSPTSTEEGTSESATP TGGTGCAACCGGCTCTCCAGGTGCTTCTCCG

ESGPGTSTEPSEGSAPG GGCACCAGCTCTACCGGTTCTCCAGGTAGC

SPAGSPTSTEEGTSTEPS CCGGCTGGCTCTCCTACCTCTACTGAGGAA

EGSAPGTSTEPSEGSAP GGTACTTCTGAAAGCGCTACTCCTGAGTCTG

GTSESATPESGPGSEPA GTCCAGGTACCTCTACTGAACCGTCCGAAG

TSGSETPGSEPATSGSET GTAGCGCTCCAGGTAGCCCAGCAGGCTCTC

PGSPAGSPTSTEEGTSES CGACTTCCACTGAGGAAGGTACTTCTACTG

ATPESGPGTSTEPSEGS AACCTTCCGAAGGCAGCGCACCAGGTACCT

APGTSTEPSEGSAPGSP CTACTGAACCTTCTGAGGGCAGCGCTCCAG

AGSPTSTEEGTSTEPSE GTACTTCTGAAAGCGCTACCCCGGAATCTG

GSAPGTSTEPSEGSAPG GCCCAGGTAGCGAACCGGCTACTTCTGGTT

TSESATPESGPGTSTEPS CTGAAACCCCAGGTAGCGAACCGGCTACCT

EGSAPGTSESATPESGP CCGGTTCTGAAACTCCAGGTAGCCCGGCAG

GSEPATSGSETPGTSTEP GCTCTCCGACCTCTACTGAGGAAGGTACTTC

SEGSAPGTSTEPSEGSA TGAAAGCGCAACCCCGGAGTCCGGCCCAGG

PGTSESATPESGPGTSES TACCTCTACCGAACCGTCTGAGGGCAGCGC

ATPESGPGSPAGSPTST ACCAGGTACTTCTACCGAACCGTCCGAGGG

EEGTSESATPESGPGSEP TAGCGCACCAGGTAGCCCAGCAGGTTCTCC hGH- SEQ SEQ XTEN Amino Acid Sequence ID DNA Nucleotide Sequence ID Name* NO: NO:

ATSGSETPGTSESATPES TACCTCCACCGAGGAAGGTACTTCTACCGA

GPGTSTEPSEGSAPGTS ACCGTCCGAGGGTAGCGCACCAGGTACCTC

TEPSEGSAPGTSTEPSEG TACTGAACCTTCTGAGGGCAGCGCTCCAGG

SAPGTSTEPSEGSAPGT TACTTCTGAAAGCGCTACCCCGGAGTCCGG

STEPSEGSAPGTSTEPSE TCCAGGTACTTCTACTGAACCGTCCGAAGG

GSAPGSPAGSPTSTEEG TAGCGCACCAGGTACTTCTGAAAGCGCAAC

TSTEPSEGSAPGTSESAT CCCTGAATCCGGTCCAGGTAGCGAACCGGC

PESGPGSEPATSGSETP TACTTCTGGCTCTGAGACTCCAGGTACTTCT

GTSESATPESGPGSEPA ACCGAACCGTCCGAAGGTAGCGCACCAGGT

TSGSETPGTSESATPESG ACTTCTACTGAACCGTCTGAAGGTAGCGCA

PGTSTEPSEGSAPGTSES CCAGGTACTTCTGAAAGCGCAACCCCGGAA

ATPESGPGSPAGSPTST TCCGGCCCAGGTACCTCTGAAAGCGCAACC

EEGSPAGSPTSTEEGSP CCGGAGTCCGGCCCAGGTAGCCCTGCTGGC

AGSPTSTEEGTSESATP TCTCCAACCTCCACCGAAGAAGGTACCTCT

ESGPGTSTEPSEGSAPG GAAAGCGCAACCCCTGAATCCGGCCCAGGT

TSESATPESGPGSEPATS AGCGAACCGGCAACCTCCGGTTCTGAAACC

GSETPGTSESATPESGP CCAGGTACCTCTGAAAGCGCTACTCCGGAG

GSEPATSGSETPGTSES TCTGGCCCAGGTACCTCTACTGAACCGTCTG

ATPESGPGTSTEPSEGS AGGGTAGCGCTCCAGGTACTTCTACTGAAC

APGSPAGSPTSTEEGTS CGTCCGAAGGTAGCGCACCAGGTACTTCTA

ESATPESGPGSEPATSG CCGAACCGTCCGAAGGCAGCGCTCCAGGTA

SETPGTSESATPESGPGS CCTCTACTGAACCTTCCGAGGGCAGCGCTC

PAGSPTSTEEGSPAGSP CAGGTACCTCTACCGAACCTTCTGAAGGTA

TSTEEGTSTEPSEGSAP GCGCACCAGGTACTTCTACCGAACCGTCCG

GTSESATPESGPGTSES AGGGTAGCGCACCAGGTAGCCCAGCAGGTT

ATPESGPGTSESATPES CTCCTACCTCCACCGAGGAAGGTACTTCTAC

GPGSEPATSGSETPGSE CGAACCGTCCGAGGGTAGCGCACCAGGTAC

PATSGSETPGSPAGSPTS CTCTGAAAGCGCAACTCCTGAGTCTGGCCC

TEEGTSTEPSEGSAPGT AGGTAGCGAACCTGCTACCTCCGGCTCTGA

STEPSEGSAPGSEPATS GACTCCAGGTACCTCTGAAAGCGCAACCCC

GSETPGTSESATPESGP GGAATCTGGTCCAGGTAGCGAACCTGCAAC

GTSTEPSEGSAPGFPTIP CTCTGGCTCTGAAACCCCAGGTACCTCTGA

LSRLFDNAMLRAHRLH AAGCGCTACTCCTGAATCTGGCCCAGGTAC

QLAFDTYQEFEEAYIPK TTCTACTGAACCGTCCGAGGGCAGCGCACC

EQKYSFLQNPQTSLCFS AGGTACTTCTGAAAGCGCTACTCCTGAGTC

ESIPTP SNREETQQKSNL CGGCCCAGGTAGCCCGGCTGGCTCTCCGAC

ELLPJSLLLIQSWLEPVQ TTCCACCGAGGAAGGTAGCCCGGCTGGCTC

FLRS VF ANSL VYG ASD S TCCAACTTCTACTGAAGAAGGTAGCCCGGC

NVYDLLKDLEEGIQTL AGGCTCTCCGACCTCTACTGAGGAAGGTAC

MGRLEDGSPRTGQIFK TTCTGAAAGCGCAACCCCGGAGTCCGGCCC

QTYSKFDTNSHNDDAL AGGTACCTCTACCGAACCGTCTGAGGGCAG

LKNYGLLYCFRKDMD CGCACCAGGTACCTCTGAAAGCGCAACTCC

KVETFLRIVQCRSVEGS TGAGTCTGGCCCAGGTAGCGAACCTGCTAC

CGFGGTSESATPESGPG CTCCGGCTCTGAGACTCCAGGTACCTCTGA

SEPATSGSETPGTSESAT AAGCGCAACCCCGGAATCTGGTCCAGGTAG

PESGPGSEPATSGSETP CGAACCTGCAACCTCTGGCTCTGAAACCCC

GTSESATPESGPGTSTEP AGGTACCTCTGAAAGCGCTACTCCTGAATC

SEGSAPGSPAGSPTSTE TGGCCCAGGTACTTCTACTGAACCGTCCGA

EGTSESATPESGPGSEP GGGCAGCGCACCAGGTAGCCCTGCTGGCTC

ATSGSETPGTSESATPES TCCAACCTCCACCGAAGAAGGTACCTCTGA

GPGSPAGSPTSTEEGSP AAGCGCAACCCCTGAATCCGGCCCAGGTAG

AGSPTSTEEGTSTEPSE CGAACCGGCAACCTCCGGTTCTGAAACCCC

GSAPGTSESATPESGPG AGGTACTTCTGAAAGCGCTACTCCTGAGTC

TSESATPESGPGTSESAT CGGCCCAGGTAGCCCGGCTGGCTCTCCGAC

PESGPGSEPATSGSETP TTCCACCGAGGAAGGTAGCCCGGCTGGCTC

GSEPATSGSETPGSPAG TCCAACTTCTACTGAAGAAGGTACTTCTACC hGH- SEQ SEQ XTEN Amino Acid Sequence ID DNA Nucleotide Sequence ID Name* NO: NO:

SPTSTEEGTSTEPSEGSA GAACCTTCCGAGGGCAGCGCACCAGGTACT PGTSTEPSEGSAPGSEP TCTGAAAGCGCTACCCCTGAGTCCGGCCCA ATSGSETPGTSESATPES GGTACTTCTGAAAGCGCTACTCCTGAATCC GPGTSTEPSEGSAPG GGTCCAGGTACTTCTGAAAGCGCTACCCCG

GAATCTGGCCCAGGTAGCGAACCGGCTACT

TCTGGTTCTGAAACCCCAGGTAGCGAACCG

GCTACCTCCGGTTCTGAAACTCCAGGTAGC

CCAGCAGGCTCTCCGACTTCCACTGAGGAA

GGTACTTCTACTGAACCTTCCGAAGGCAGC

GCACCAGGTACCTCTACTGAACCTTCTGAG

GGCAGCGCTCCAGGTAGCGAACCTGCAACC

TCTGGCTCTGAAACCCCAGGTACCTCTGAA

AGCGCTACTCCTGAATCTGGCCCAGGTACTT

CTACTGAACCGTCCGAGGGCAGCGCACCAG

GTTTTCCGACTATTCCGCTGTCTCGTCTGTTT

GATAATGCTATGCTGCGTGCGCACCGTCTG

CACCAGCTGGCCTTTGATACTTACCAGGAA

TTTGAAGAAGCcTACATTCCTAAAGAGCAG

AAGTACTCTTTCCTGCAAAACCCACAGACTT

CTCTCTGCTTCAGCGAATCTATTCCGACGCC

TTCCAATCGCGAGGAAACTCAGCAAAAGTC

CAATCTGGAACTACTCCGCATTTCTCTGCTT

CTGATTCAGAGCTGGCTAGAACCAGTGCAA

TTTCTGCGTTCCGTCTTCGCCAATAGCCTAG

TTTATGGCGCATCCGACAGCAACGTATACG

ATCTCCTGAAAGATCTCGAGGAAGGCATTC

AGACCCTGATGGGTCGTCTCGAGGATGGCT

CTCCGCGTACTGGTCAGATCTTCAAGCAGA

CTTACTCTAAATTTGATACTAACAGCCACAA

TGACGATGCGCTTCTAAAAAACTATGGTCT

GCTGTATTGTTTTCGTAAAGATATGGACAA

AGTTGAAACCTTCCTGCGTATTGTTCAGTGT

CGTTCCGTTGAGGGCAGCTGTGGTTTCTAAG

GTGGTACCTCTGAAAGCGCAACTCCTGAGT

CTGGCCCAGGTAGCGAACCTGCTACCTCCG

GCTCTGAGACTCCAGGTACCTCTGAAAGCG

CAACCCCGGAATCTGGTCCAGGTAGCGAAC

CTGCAACCTCTGGCTCTGAAACCCCAGGTA

CCTCTGAAAGCGCTACTCCTGAATCTGGCCC

AGGTACTTCTACTGAACCGTCCGAGGGCAG

CGCACCAGGTAGCCCTGCTGGCTCTCCAAC

CTCCACCGAAGAAGGTACCTCTGAAAGCGC

AACCCCTGAATCCGGCCCAGGTAGCGAACC

GGCAACCTCCGGTTCTGAAACCCCAGGTAC

TTCTGAAAGCGCTACTCCTGAGTCCGGCCC

AGGTAGCCCGGCTGGCTCTCCGACTTCCAC

CGAGGAAGGTAGCCCGGCTGGCTCTCCAAC

TTCTACTGAAGAAGGTACTTCTACCGAACCT

TCCGAGGGCAGCGCACCAGGTACTTCTGAA

AGCGCTACCCCTGAGTCCGGCCCAGGTACT

TCTGAAAGCGCTACTCCTGAATCCGGTCCA

GGTACTTCTGAAAGCGCTACCCCGGAATCT

GGCCCAGGTAGCGAACCGGCTACTTCTGGT

TCTGAAACCCCAGGTAGCGAACCGGCTACC

TCCGGTTCTGAAACTCCAGGTAGCCCAGCA

GGCTCTCCGACTTCCACTGAGGAAGGTACT hGH- SEQ SEQ XTEN Amino Acid Sequence ID DNA Nucleotide Sequence ID Name* NO: NO:

TCTACTGAACCTTCCGAAGGCAGCGCACCA

GGTACCTCTACTGAACCTTCTGAGGGCAGC

GCTCCAGGTAGCGAACCTGCAACCTCTGGC

TCTGAAACCCCAGGTACCTCTGAAAGCGCT

ACTCCTGAATCTGGCCCAGGTACTTCTACTG

AACCGTCCGAGGGCAGCGCACCA

AM875- GTSTEPSEGSAPGSEPA 6 GGTACTTCTACTGAACCGTCTGAAGGCAGC 12 hGH TSGSETPGSPAGSPTSTE GCACCAGGTAGCGAACCGGCTACTTCCGGT

EGSTSSTAESPGPGTSTP TCTGAAACCCCAGGTAGCCCAGCAGGTTCT

ESGSASPGSTSESPSGTA CCAACTTCTACTGAAGAAGGTTCTACCAGC

PGSTSESPSGTAPGTSTP TCTACCGCAGAATCTCCTGGTCCAGGTACCT

ESGSASPGTSTPESGSAS CTACTCCGGAAAGCGGCTCTGCATCTCCAG

PGSEPATSGSETPGTSES GTTCTACTAGCGAATCTCCTTCTGGCACTGC

ATPESGPGSPAGSPTST ACCAGGTTCTACTAGCGAATCCCCGTCTGGT

EEGTSTEPSEGSAPGTS ACTGCTCCAGGTACTTCTACTCCTGAAAGCG

ESATPESGPGTSTEPSEG GTTCCGCTTCTCCAGGTACCTCTACTCCGGA

SAPGTSTEPSEGSAPGSP AAGCGGTTCTGCATCTCCAGGTAGCGAACC

AGSPTSTEEGTSTEPSE GGCAACCTCCGGCTCTGAAACCCCAGGTAC

GSAPGTSTEPSEGSAPG CTCTGAAAGCGCTACTCCTGAATCCGGCCC

TSESATPESGPGTSESAT AGGTAGCCCGGCAGGTTCTCCGACTTCCAC

PESGPGTSTEPSEGSAP TGAGGAAGGTACCTCTACTGAACCTTCTGA

GTSTEPSEGSAPGTSES GGGCAGCGCTCCAGGTACTTCTGAAAGCGC

ATPESGPGTSTEPSEGS TACCCCGGAGTCCGGTCCAGGTACTTCTACT

APGSEPATSGSETPGSP GAACCGTCCGAAGGTAGCGCACCAGGTACT

AGSPTSTEEGSSTPSGA TCTACCGAACCGTCCGAGGGTAGCGCACCA

TGSPGTPGSGTASSSPG GGTAGCCCAGCAGGTTCTCCTACCTCCACC

SSTPSGATGSPGTSTEPS GAGGAAGGTACTTCTACCGAACCGTCCGAG

EGSAPGTSTEPSEGSAP GGTAGCGCACCAGGTACTTCTACCGAACCT

GSEPATSGSETPGSPAG TCCGAGGGCAGCGCACCAGGTACTTCTGAA

SPTSTEEGSPAGSPTSTE AGCGCTACCCCTGAGTCCGGCCCAGGTACT

EGTSTEPSEGSAPGASA TCTGAAAGCGCTACTCCTGAATCCGGTCCA

SGAPSTGGTSESATPES GGTACCTCTACTGAACCTTCCGAAGGCAGC

GPGSPAGSPTSTEEGSP GCTCCAGGTACCTCTACCGAACCGTCCGAG

AGSPTSTEEGSTSSTAES GGCAGCGCACCAGGTACTTCTGAAAGCGCA

PGPGSTSESPSGTAPGTS ACCCCTGAATCCGGTCCAGGTACTTCTACTG

PSGESSTAPGTPGSGTA AACCTTCCGAAGGTAGCGCTCCAGGTAGCG

SSSPGSSTPSGATGSPGS AACCTGCTACTTCTGGTTCTGAAACCCCAGG

SPSASTGTGPGSEPATS TAGCCCGGCTGGCTCTCCGACCTCCACCGA

GSETPGTSESATPESGP GGAAGGTAGCTCTACCCCGTCTGGTGCTAC

GSEPATSGSETPGSTSST TGGTTCTCCAGGTACTCCGGGCAGCGGTAC

AESPGPGSTSSTAESPGP TGCTTCTTCCTCTCCAGGTAGCTCTACCCCT

GTSPSGESSTAPGSEPA TCTGGTGCTACTGGCTCTCCAGGTACCTCTA

TSGSETPGSEPATSGSET CCGAACCGTCCGAGGGTAGCGCACCAGGTA

PGTSTEPSEGSAPGSTSS CCTCTACTGAACCGTCTGAGGGTAGCGCTC

TAESPGPGTSTPESGSA CAGGTAGCGAACCGGCAACCTCCGGTTCTG

SPGSTSESPSGTAPGTST AAACTCCAGGTAGCCCTGCTGGCTCTCCGA

EPSEGSAPGTSTEPSEGS CTTCTACTGAGGAAGGTAGCCCGGCTGGTT

APGTSTEPSEGSAPGSS CTCCGACTTCTACTGAGGAAGGTACTTCTAC

TPSGATGSPGSSPSAST CGAACCTTCCGAAGGTAGCGCTCCAGGTGC

GTGPGASPGTSSTGSPG AAGCGCAAGCGGCGCGCCAAGCACGGGAG

SEPATSGSETPGTSESAT GTACTTCTGAAAGCGCTACTCCTGAGTCCG

PESGPGSPAGSPTSTEE GCCCAGGTAGCCCGGCTGGCTCTCCGACTT

GSSTPSGATGSPGSSPS CCACCGAGGAAGGTAGCCCGGCTGGCTCTC

ASTGTGPGASPGTSSTG CAACTTCTACTGAAGAAGGTTCTACCAGCT

SPGTSESATPESGPGTST CTACCGCTGAATCTCCTGGCCCAGGTTCTAC

EPSEGSAPGTSTEPSEGS TAGCGAATCTCCGTCTGGCACCGCACCAGG hGH- SEQ SEQ XTEN Amino Acid Sequence ID DNA Nucleotide Sequence ID Name* NO: NO:

APGFPTIPLSRLFDNAM TACTTCCCCTAGCGGTGAATCTTCTACTGCA

LRAHRLHQLAFDTYQE CCAGGTACCCCTGGCAGCGGTACCGCTTCTT

FEEAYIPKEQKYSFLQN CCTCTCCAGGTAGCTCTACCCCGTCTGGTGC

PQTSLCFSESIPTPSNRE TACTGGCTCTCCAGGTTCTAGCCCGTCTGCA

ETQQKSNLELLRISLLLI TCTACCGGTACCGGCCCAGGTAGCGAACCG

QSWLEPVQFLRSVFAN GCAACCTCCGGCTCTGAAACTCCAGGTACT

SL VYG ASD SNVYDLLK TCTGAAAGCGCTACTCCGGAATCCGGCCCA

DLEEGIQTLMGRLEDGS GGTAGCGAACCGGCTACTTCCGGCTCTGAA

PRTGQIFKQTYSKFDTN ACCCCAGGTTCCACCAGCTCTACTGCAGAA

SHNDDALLKNYGLLYC TCTCCGGGCCCAGGTTCTACTAGCTCTACTG

FRKDMDKVETFLRIVQ CAGAATCTCCGGGTCCAGGTACTTCTCCTAG

CRSVEGSCGF CGGCGAATCTTCTACCGCTCCAGGTAGCGA

ACCGGCAACCTCTGGCTCTGAAACTCCAGG

TAGCGAACCTGCAACCTCCGGCTCTGAAAC

CCCAGGTACTTCTACTGAACCTTCTGAGGGC

AGCGCACCAGGTTCTACCAGCTCTACCGCA

GAATCTCCTGGTCCAGGTACCTCTACTCCGG

AAAGCGGCTCTGCATCTCCAGGTTCTACTA

GCGAATCTCCTTCTGGCACTGCACCAGGTA

CTTCTACCGAACCGTCCGAAGGCAGCGCTC

CAGGTACCTCTACTGAACCTTCCGAGGGCA

GCGCTCCAGGTACCTCTACCGAACCTTCTGA

AGGTAGCGCACCAGGTAGCTCTACTCCGTC

TGGTGCAACCGGCTCCCCAGGTTCTAGCCC

GTCTGCTTCCACTGGTACTGGCCCAGGTGCT

TCCCCGGGCACCAGCTCTACTGGTTCTCCAG

GTAGCGAACCTGCTACCTCCGGTTCTGAAA

CCCCAGGTACCTCTGAAAGCGCAACTCCGG

AGTCTGGTCCAGGTAGCCCTGCAGGTTCTCC

TACCTCCACTGAGGAAGGTAGCTCTACTCC

GTCTGGTGCAACCGGCTCCCCAGGTTCTAG

CCCGTCTGCTTCCACTGGTACTGGCCCAGGT

GCTTCCCCGGGCACCAGCTCTACTGGTTCTC

CAGGTACCTCTGAAAGCGCTACTCCGGAGT

CTGGCCCAGGTACCTCTACTGAACCGTCTG

AGGGTAGCGCTCCAGGTACTTCTACTGAAC

CGTCCGAAGGTAGCGCACCAGGTTTTCCGA

CTATTCCGCTGTCTCGTCTGTTTGATAATGC

TATGCTGCGTGCGCACCGTCTGCACCAGCT

GGCCTTTGATACTTACCAGGAATTTGAAGA

AGCcTACATTCCTAAAGAGCAGAAGTACTCT

TTCCTGCAAAACCCACAGACTTCTCTCTGCT

TCAGCGAATCTATTCCGACGCCTTCCAATCG

CGAGGAAACTCAGCAAAAGTCCAATCTGGA

ACTACTCCGCATTTCTCTGCTTCTGATTCAG

AGCTGGCTAGAACCAGTGCAATTTCTGCGT

TCCGTCTTCGCCAATAGCCTAGTTTATGGCG

CATCCGACAGCAACGTATACGATCTCCTGA

AAGATCTCGAGGAAGGCATTCAGACCCTGA

TGGGTCGTCTCGAGGATGGCTCTCCGCGTA

CTGGTCAGATCTTCAAGCAGACTTACTCTAA

ATTTGATACTAACAGCCACAATGACGATGC

GCTTCTAAAAAACTATGGTCTGCTGTATTGT

TTTCGTAAAGATATGGACAAAGTTGAAACC

TTCCTGCGTATTGTTCAGTGTCGTTCCGTTG

AGGGCAGCTGTGGTTTCTAA As described more fully in, for example, W013/184216, WO/2014/164568, U.S. Patent Nos. 8,492,530 and 8,703,717, each of which is incorporated herein by reference in its entirety, the fusion proteins optionally include spacer sequences that further comprise cleavage sequences to release the hGH from the fusion protein when acted on by a protease, releasing hGH from the XTEN sequence(s).

VI). EXTENDED RECOMBINANT POLYPEPTIDES

The present invention comprises formulations comprising XTEN polypeptide compositions that are useful as a fusion protein partner to which hGH is linked, resulting in a hGH-XTEN fusion protein. XTEN are generally extended length polypeptides with non- naturally occurring, substantially non-repetitive sequences that are composed mainly of small hydrophilic amino acids, with the sequence having a low degree or no secondary or tertiary structure under physiologic conditions. Among other characteristics, features of XTEN sequences such as the non-repetitiveness of sequences, exemplary sequence motifs, and lengths of sequences are known in the art and are described at least in, for example,

W013/184216, WO/2014/164568 and U.S. Patent Nos. 8,492,530 and 8,703,717, each of which is incorporated herein by reference in its entirety.

In one embodiment, the hGH-XTEN of the formulation can comprise one or more XTEN sequence wherein the sequence exhibits at least about 80% sequence identity, or alternatively 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to a XTEN selected from Table 2. Examples where more than one XTEN is used in a hGH-XTEN composition include, but are not limited to constructs with an XTEN linked to both the N- and C-termini of at least one hGH. In preferred embodiments of the present invention comprising constructs with an

XTEN linked to both the N- and C-termini of at least one hGH, the length of the N-terminal XTEN sequence ranges between about 500 amino acids to about 1320 amino acids and the length of the C-terminal XTEN ranges between about 40 amino acids to about 300 amino acids.

Table 2: XTEN Polypeptides

SEQ

XTEN ID Amino Acid Sequence

Name

NO:

AM48 14 MAEPAGSPTSTEEGASPGTSSTGSPGSSTPSGATGSPGSSTPSGATGS

AE144 15 GSEPATSGSETPGTSESATPESGPGSEPATSGSETPGSPAGSPTSTEEGTSTEPSEGS

APGSEPATSGSETPGSEPATSGSETPGSEPATSGSETPGTSTEPSEGSAPGTSESATP ESGPGSEPATSGSETPGTSTEPSEGSAP

AF144 16 GTSTPESGSASPGTSPSGESSTAPGTSPSGESSTAPGSTSSTAESPGPGSTSESPSGTA

PGSTSSTAESPGPGTSPSGESSTAPGTSTPESGSASPGSTSSTAESPGPGTSPSGESST APGTSPSGESSTAPGTSPSGESSTAP

AE288 17 GTSESATPESGPGSEPATSGSETPGTSESATPESGPGSEPATSGSETPGTSESATPES

GPGTSTEPSEGSAPGSPAGSPTSTEEGTSESATPESGPGSEPATSGSETPGTSESATP ESGPGSPAGSPTSTEEGSPAGSPTSTEEGTSTEPSEGSAPGTSESATPESGPGTSESA TPESGPGTSESATPESGPGSEPATSGSETPGSEPATSGSETPGSPAGSPTSTEEGTST EPSEGSAPGTSTEPSEGSAPGSEPATSGSETPGTSESATPESGPGTSTEPSEGSAP

AF504 18 GASPGTSSTGSPGSSPSASTGTGPGSSPSASTGTGPGTPGSGTASSSPGSSTPSGATG

SPGSXPSASTGTGPGASPGTSSTGSPGTPGSGTASSSPGSSTPSGATGSPGTPGSGT

ASSSPGASPGTSSTGSPGASPGTSSTGSPGTPGSGTASSSPGSSTPSGATGSPGASPG

TSSTGSPGTPGSGTASSSPGSSTPSGATGSPGSXPSASTGTGPGSSPSASTGTGPGSS

TPSGATGSPGSSTPSGATGSPGASPGTSSTGSPGASPGTSSTGSPGASPGTSSTGSPG

TPGSGTASSSPGASPGTSSTGSPGASPGTSSTGSPGASPGTSSTGSPGSSPSASTGTG

PGTPGSGTASSSPGASPGTSSTGSPGASPGTSSTGSPGASPGTSSTGSPGSSTPSGAT

GSPGSSTPSGATGSPGASPGTSSTGSPGTPGSGTASSSPGSSTPSGATGSPGSSTPSG

ATGSPGSSTPSGATGSPGSSPSASTGTGPGASPGTSSTGSP

AF540 19 GSTSSTAESPGPGSTSSTAESPGPGSTSESPSGTAPGSTSSTAESPGPGSTSSTAESPG

PGTSTPESGSASPGSTSESPSGTAPGTSPSGESSTAPGSTSESPSGTAPGSTSESPSGT

APGTSPSGESSTAPGSTSESPSGTAPGSTSESPSGTAPGTSPSGESSTAPGSTSESPSG

TAPGSTSESPSGTAPGSTSESPSGTAPGTSTPESGSASPGSTSESPSGTAPGTSTPESG

SASPGSTSSTAESPGPGSTSSTAESPGPGTSTPESGSASPGTSTPESGSASPGSTSESP

SGTAPGTSTPESGSASPGTSTPESGSASPGSTSESPSGTAPGSTSESPSGTAPGSTSES

PSGTAPGSTSSTAESPGPGTSTPESGSASPGTSTPESGSASPGSTSESPSGTAPGSTSE

SPSGTAPGTSTPESGSASPGSTSESPSGTAPGSTSESPSGTAPGTSTPESGSASPGTSP

SGESSTAPGSTSSTAESPGPGTSPSGESSTAPGSTSSTAESPGPGTSTPESGSASPGST

SESPSGTAP

AD576 20 GSSESGSSEGGPGSGGEPSESGSSGSSESGSSEGGPGSSESGSSEGGPGSSESGSSEG

GPGSSESGSSEGGPGSSESGSSEGGPGESPGGSSGSESGSEGSSGPGESSGSSESGSS

EGGPGSSESGSSEGGPGSSESGSSEGGPGSGGEPSESGSSGESPGGSSGSESGESPG

GSSGSESGSGGEPSESGSSGSSESGSSEGGPGSGGEPSESGSSGSGGEPSESGSSGSE

GSSGPGESSGESPGGSSGSESGSGGEPSESGSSGSGGEPSESGSSGSGGEPSESGSSG

SSESGSSEGGPGESPGGSSGSESGESPGGSSGSESGESPGGSSGSESGESPGGSSGSE

SGESPGGSSGSESGSSESGSSEGGPGSGGEPSESGSSGSEGSSGPGESSGSSESGSSE

GGPGSGGEPSESGSSGSSESGSSEGGPGSGGEPSESGSSGESPGGSSGSESGESPGG

SSGSESGSSESGSSEGGPGSGGEPSESGSSGSSESGSSEGGPGSGGEPSESGSSGSGG

EPSESGSSGESPGGSSGSESGSEGSSGPGESSGSSESGSSEGGPGSEGSSGPGESS

AE576 21 GSPAGSPTSTEEGTSESATPESGPGTSTEPSEGSAPGSPAGSPTSTEEGTSTEPSEGS

APGTSTEPSEGSAPGTSESATPESGPGSEPATSGSETPGSEPATSGSETPGSPAGSPT

STEEGTSESATPESGPGTSTEPSEGSAPGTSTEPSEGSAPGSPAGSPTSTEEGTSTEPS

EGSAPGTSTEPSEGSAPGTSESATPESGPGTSTEPSEGSAPGTSESATPESGPGSEPA

TSGSETPGTSTEPSEGSAPGTSTEPSEGSAPGTSESATPESGPGTSESATPESGPGSP

AGSPTSTEEGTSESATPESGPGSEPATSGSETPGTSESATPESGPGTSTEPSEGSAPG

TSTEPSEGSAPGTSTEPSEGSAPGTSTEPSEGSAPGTSTEPSEGSAPGTSTEPSEGSA

PGSPAGSPTSTEEGTSTEPSEGSAPGTSESATPESGPGSEPATSGSETPGTSESATPE

SGPGSEPATSGSETPGTSESATPESGPGTSTEPSEGSAPGTSESATPESGPGSPAGSP

TSTEEGSPAGSPTSTEEGSPAGSPTSTEEGTSESATPESGPGTSTEPSEGSAP

AF576 22 GSTSSTAESPGPGSTSSTAESPGPGSTSESPSGTAPGSTSSTAESPGPGSTSSTAESPG

PGTSTPESGSASPGSTSESPSGTAPGTSPSGESSTAPGSTSESPSGTAPGSTSESPSGT APGTSPSGESSTAPGSTSESPSGTAPGSTSESPSGTAPGTSPSGESSTAPGSTSESPSG TAPGSTSESPSGTAPGSTSESPSGTAPGTSTPESGSASPGSTSESPSGTAPGTSTPESG SASPGSTSSTAESPGPGSTSSTAESPGPGTSTPESGSASPGTSTPESGSASPGSTSESP SEQ

XTEN ID Amino Acid Sequence

Name

NO:

SGTAPGTSTPESGSASPGTSTPESGSASPGSTSESPSGTAPGSTSESPSGTAPGSTSES PSGTAPGSTSSTAESPGPGTSTPESGSASPGTSTPESGSASPGSTSESPSGTAPGSTSE SPSGTAPGTSTPESGSASPGSTSESPSGTAPGSTSESPSGTAPGTSTPESGSASPGTSP SGESSTAPGSTSSTAESPGPGTSPSGESSTAPGSTSSTAESPGPGTSTPESGSASPGST SESPSGTAPGSTSSTAESPGPGTSTPESGSASPGTSTPESGSASP

AE624 23 MAEPAGSPTSTEEGTPGSGTASSSPGSSTPSGATGSPGASPGTSSTGSPGSPAGSPT

STEEGTSESATPESGPGTSTEPSEGSAPGSPAGSPTSTEEGTSTEPSEGSAPGTSTEPS

EGSAPGTSESATPESGPGSEPATSGSETPGSEPATSGSETPGSPAGSPTSTEEGTSES

ATPESGPGTSTEPSEGSAPGTSTEPSEGSAPGSPAGSPTSTEEGTSTEPSEGSAPGTS

TEPSEGSAPGTSESATPESGPGTSTEPSEGSAPGTSESATPESGPGSEPATSGSETPG

TSTEPSEGSAPGTSTEPSEGSAPGTSESATPESGPGTSESATPESGPGSPAGSPTSTE

EGTSESATPESGPGSEPATSGSETPGTSESATPESGPGTSTEPSEGSAPGTSTEPSEG

SAPGTSTEPSEGSAPGTSTEPSEGSAPGTSTEPSEGSAPGTSTEPSEGSAPGSPAGSP

TSTEEGTSTEPSEGSAPGTSESATPESGPGSEPATSGSETPGTSESATPESGPGSEPA

TSGSETPGTSESATPESGPGTSTEPSEGSAPGTSESATPESGPGSPAGSPTSTEEGSP

AGSPTSTEEGSPAGSPTSTEEGTSESATPESGPGTSTEPSEGSAP

AD836 24 GSSESGSSEGGPGSSESGSSEGGPGESPGGSSGSESGSGGEPSESGSSGESPGGSSGS

ESGESPGGSSGSESGSSESGSSEGGPGSSESGSSEGGPGSSESGSSEGGPGESPGGSS

GSESGESPGGSSGSESGESPGGSSGSESGSSESGSSEGGPGSSESGSSEGGPGSSESG

SSEGGPGSSESGSSEGGPGSSESGSSEGGPGSSESGSSEGGPGSGGEPSESGSSGESP

GGSSGSESGESPGGSSGSESGSGGEPSESGSSGSEGSSGPGESSGSSESGSSEGGPGS

GGEPSESGSSGSEGSSGPGESSGSSESGSSEGGPGSGGEPSESGSSGESPGGSSGSES

GSGGEPSESGSSGSGGEPSESGSSGSSESGSSEGGPGSGGEPSESGSSGSGGEPSESG

SSGSEGSSGPGESSGESPGGSSGSESGSEGSSGPGESSGSEGSSGPGESSGSGGEPSE

SGSSGSSESGSSEGGPGSSESGSSEGGPGESPGGSSGSESGSGGEPSESGSSGSEGSS

GPGESSGESPGGSSGSESGSEGSSGPGSSESGSSEGGPGSGGEPSESGSSGSEGSSGP

GESSGSEGSSGPGESSGSEGSSGPGESSGSGGEPSESGSSGSGGEPSESGSSGESPGG

SSGSESGESPGGSSGSESGSGGEPSESGSSGSEGSSGPGESSGESPGGSSGSESGSSE

SGSSEGGPGSSESGSSEGGPGSSESGSSEGGPGSGGEPSESGSSGSSESGSSEGGPGE

SPGGSSGSESGSGGEPSESGSSGSSESGSSEGGPGESPGGSSGSESGSGGEPSESGSS

GESPGGSSGSESGSGGEPSESGSS

AE864 25 GSPAGSPTSTEEGTSESATPESGPGTSTEPSEGSAPGSPAGSPTSTEEGTSTEPSEGS

APGTSTEPSEGSAPGTSESATPESGPGSEPATSGSETPGSEPATSGSETPGSPAGSPT

STEEGTSESATPESGPGTSTEPSEGSAPGTSTEPSEGSAPGSPAGSPTSTEEGTSTEPS

EGSAPGTSTEPSEGSAPGTSESATPESGPGTSTEPSEGSAPGTSESATPESGPGSEPA

TSGSETPGTSTEPSEGSAPGTSTEPSEGSAPGTSESATPESGPGTSESATPESGPGSP

AGSPTSTEEGTSESATPESGPGSEPATSGSETPGTSESATPESGPGTSTEPSEGSAPG

TSTEPSEGSAPGTSTEPSEGSAPGTSTEPSEGSAPGTSTEPSEGSAPGTSTEPSEGSA

PGSPAGSPTSTEEGTSTEPSEGSAPGTSESATPESGPGSEPATSGSETPGTSESATPE

SGPGSEPATSGSETPGTSESATPESGPGTSTEPSEGSAPGTSESATPESGPGSPAGSP

TSTEEGSPAGSPTSTEEGSPAGSPTSTEEGTSESATPESGPGTSTEPSEGSAPGTSES

ATPESGPGSEPATSGSETPGTSESATPESGPGSEPATSGSETPGTSESATPESGPGTS

TEPSEGSAPGSPAGSPTSTEEGTSESATPESGPGSEPATSGSETPGTSESATPESGPG

SPAGSPTSTEEGSPAGSPTSTEEGTSTEPSEGSAPGTSESATPESGPGTSESATPESG

PGTSESATPESGPGSEPATSGSETPGSEPATSGSETPGSPAGSPTSTEEGTSTEPSEG

SAPGTSTEPSEGSAPGSEPATSGSETPGTSESATPESGPGTSTEPSEGSAP

AF864 26 GSTSESPSGTAPGTSPSGESSTAPGSTSESPSGTAPGSTSESPSGTAPGTSTPESGSAS

PGTSTPESGSASPGSTSESPSGTAPGSTSESPSGTAPGTSPSGESSTAPGSTSESPSGT

APGTSPSGESSTAPGTSPSGESSTAPGSTSSTAESPGPGTSPSGESSTAPGTSPSGESS

TAPGSTSSTAESPGPGTSTPESGSASPGTSTPESGSASPGSTSESPSGTAPGSTSESPS

GTAPGTSTPESGSASPGSTSSTAESPGPGTSTPESGSASPGSTSESPSGTAPGTSPSG

ESSTAPGSTSSTAESPGPGTSPSGESSTAPGTSTPESGSASPGSTSSTAESPGPGSTSS

TAESPGPGSTSSTAESPGPGSTSSTAESPGPGTSPSGESSTAPGSTSESPSGTAPGSTS

ESPSGTAPGTSTPESGPXXXGASASGAPSTXXXXSESPSGTAPGSTSESPSGTAPGS

TSESPSGTAPGSTSESPSGTAPGSTSESPSGTAPGSTSESPSGTAPGTSTPESGSASPG

TSPSGESSTAPGTSPSGESSTAPGSTSSTAESPGPGTSPSGESSTAPGTSTPESGSASP

GSTSESPSGTAPGSTSESPSGTAPGTSPSGESSTAPGSTSESPSGTAPGTSTPESGSAS SEQ

XTEN ID Amino Acid Sequence

Name

NO:

PGTSTPESGSASPGSTSESPSGTAPGTSTPESGSASPGSTSSTAESPGPGSTSESPSGT APGSTSESPSGTAPGTSPSGESSTAPGSTSSTAESPGPGTSPSGESSTAPGTSTPESGS ASPGTSPSGESSTAPGTSPSGESSTAPGTSPSGESSTAPGSTSSTAESPGPGSTSSTAE SPGPGTSPSGESSTAPGSSPSASTGTGPGSSTPSGATGSPGSSTPSGATGSP

AG864 27 GASPGTSSTGSPGSSPSASTGTGPGSSPSASTGTGPGTPGSGTASSSPGSSTPSGATG

SPGSSPSASTGTGPGASPGTSSTGSPGTPGSGTASSSPGSSTPSGATGSPGTPGSGTA

SSSPGASPGTSSTGSPGASPGTSSTGSPGTPGSGTASSSPGSSTPSGATGSPGASPGT

SSTGSPGTPGSGTASSSPGSSTPSGATGSPGSSPSASTGTGPGSSPSASTGTGPGSST

PSGATGSPGSSTPSGATGSPGASPGTSSTGSPGASPGTSSTGSPGASPGTSSTGSPGT

PGSGTASSSPGASPGTSSTGSPGASPGTSSTGSPGASPGTSSTGSPGSSPSASTGTGP

GTPGSGTASSSPGASPGTSSTGSPGASPGTSSTGSPGASPGTSSTGSPGSSTPSGATG

SPGSSTPSGATGSPGASPGTSSTGSPGTPGSGTASSSPGSSTPSGATGSPGSSTPSGA

TGSPGSSTPSGATGSPGSSPSASTGTGPGASPGTSSTGSPGASPGTSSTGSPGTPGSG

TASSSPGASPGTSSTGSPGASPGTSSTGSPGASPGTSSTGSPGASPGTSSTGSPGTPG

SGTASSSPGSSTPSGATGSPGTPGSGTASSSPGSSTPSGATGSPGTPGSGTASSSPGS

STPSGATGSPGSSTPSGATGSPGSSPSASTGTGPGSSPSASTGTGPGASPGTSSTGSP

GTPGSGTASSSPGSSTPSGATGSPGSSPSASTGTGPGSSPSASTGTGPGASPGTSSTG

SPGASPGTSSTGSPGSSTPSGATGSPGSSPSASTGTGPGASPGTSSTGSPGSSPSAST

GTGPGTPGSGTASSSPGSSTPSGATGSPGSSTPSGATGSPGASPGTSSTGSP

AM87 28 GTSTEPSEGSAPGSEPATSGSETPGSPAGSPTSTEEGSTSSTAESPGPGTSTPESGSA 5 SPGSTSESPSGTAPGSTSESPSGTAPGTSTPESGSASPGTSTPESGSASPGSEPATSGS

ETPGTSESATPESGPGSPAGSPTSTEEGTSTEPSEGSAPGTSESATPESGPGTSTEPSE

GSAPGTSTEPSEGSAPGSPAGSPTSTEEGTSTEPSEGSAPGTSTEPSEGSAPGTSESA

TPESGPGTSESATPESGPGTSTEPSEGSAPGTSTEPSEGSAPGTSESATPESGPGTST

EPSEGSAPGSEPATSGSETPGSPAGSPTSTEEGSSTPSGATGSPGTPGSGTASSSPGS

STPSGATGSPGTSTEPSEGSAPGTSTEPSEGSAPGSEPATSGSETPGSPAGSPTSTEE

GSPAGSPTSTEEGTSTEPSEGSAPGASASGAPSTGGTSESATPESGPGSPAGSPTSTE

EGSPAGSPTSTEEGSTSSTAESPGPGSTSESPSGTAPGTSPSGESSTAPGTPGSGTAS

SSPGSSTPSGATGSPGSSPSASTGTGPGSEPATSGSETPGTSESATPESGPGSEPATS

GSETPGSTSSTAESPGPGSTSSTAESPGPGTSPSGESSTAPGSEPATSGSETPGSEPA

TSGSETPGTSTEPSEGSAPGSTSSTAESPGPGTSTPESGSASPGSTSESPSGTAPGTST

EPSEGSAPGTSTEPSEGSAPGTSTEPSEGSAPGSSTPSGATGSPGSSPSASTGTGPGA

SPGTSSTGSPGSEPATSGSETPGTSESATPESGPGSPAGSPTSTEEGSSTPSGATGSP

GSSPSASTGTGPGASPGTSSTGSPGTSESATPESGPGTSTEPSEGSAPGTSTEPSEGS

AP

AE912 29 MAEPAGSPTSTEEGTPGSGTASSSPGSSTPSGATGSPGASPGTSSTGSPGSPAGSPT

STEEGTSESATPESGPGTSTEPSEGSAPGSPAGSPTSTEEGTSTEPSEGSAPGTSTEPS

EGSAPGTSESATPESGPGSEPATSGSETPGSEPATSGSETPGSPAGSPTSTEEGTSES

ATPESGPGTSTEPSEGSAPGTSTEPSEGSAPGSPAGSPTSTEEGTSTEPSEGSAPGTS

TEPSEGSAPGTSESATPESGPGTSTEPSEGSAPGTSESATPESGPGSEPATSGSETPG

TSTEPSEGSAPGTSTEPSEGSAPGTSESATPESGPGTSESATPESGPGSPAGSPTSTE

EGTSESATPESGPGSEPATSGSETPGTSESATPESGPGTSTEPSEGSAPGTSTEPSEG

SAPGTSTEPSEGSAPGTSTEPSEGSAPGTSTEPSEGSAPGTSTEPSEGSAPGSPAGSP

TSTEEGTSTEPSEGSAPGTSESATPESGPGSEPATSGSETPGTSESATPESGPGSEPA

TSGSETPGTSESATPESGPGTSTEPSEGSAPGTSESATPESGPGSPAGSPTSTEEGSP

AGSPTSTEEGSPAGSPTSTEEGTSESATPESGPGTSTEPSEGSAPGTSESATPESGPG

SEPATSGSETPGTSESATPESGPGSEPATSGSETPGTSESATPESGPGTSTEPSEGSA

PGSPAGSPTSTEEGTSESATPESGPGSEPATSGSETPGTSESATPESGPGSPAGSPTS

TEEGSPAGSPTSTEEGTSTEPSEGSAPGTSESATPESGPGTSESATPESGPGTSESAT

PESGPGSEPATSGSETPGSEPATSGSETPGSPAGSPTSTEEGTSTEPSEGSAPGTSTE

PSEGSAPGSEPATSGSETPGTSESATPESGPGTSTEPSEGSAP

AM92 30 MAEPAGSPTSTEEGASPGTSSTGSPGSSTPSGATGSPGSSTPSGATGSPGTSTEPSE 3 GSAPGSEPATSGSETPGSPAGSPTSTEEGSTSSTAESPGPGTSTPESGSASPGSTSESP

SGTAPGSTSESPSGTAPGTSTPESGSASPGTSTPESGSASPGSEPATSGSETPGTSES

ATPESGPGSPAGSPTSTEEGTSTEPSEGSAPGTSESATPESGPGTSTEPSEGSAPGTS

TEPSEGSAPGSPAGSPTSTEEGTSTEPSEGSAPGTSTEPSEGSAPGTSESATPESGPG

TSESATPESGPGTSTEPSEGSAPGTSTEPSEGSAPGTSESATPESGPGTSTEPSEGSA SEQ

XTEN ID Amino Acid Sequence

Name

NO:

PGSEPATSGSETPGSPAGSPTSTEEGSSTPSGATGSPGTPGSGTASSSPGSSTPSGAT

GSPGTSTEPSEGSAPGTSTEPSEGSAPGSEPATSGSETPGSPAGSPTSTEEGSPAGSP

TSTEEGTSTEPSEGSAPGASASGAPSTGGTSESATPESGPGSPAGSPTSTEEGSPAGS

PTSTEEGSTSSTAESPGPGSTSESPSGTAPGTSPSGESSTAPGTPGSGTASSSPGSSTP

SGATGSPGSSPSASTGTGPGSEPATSGSETPGTSESATPESGPGSEPATSGSETPGST

SSTAESPGPGSTSSTAESPGPGTSPSGESSTAPGSEPATSGSETPGSEPATSGSETPG

TSTEPSEGSAPGSTSSTAESPGPGTSTPESGSASPGSTSESPSGTAPGTSTEPSEGSAP

GTSTEPSEGSAPGTSTEPSEGSAPGSSTPSGATGSPGSSPSASTGTGPGASPGTSSTG

SPGSEPATSGSETPGTSESATPESGPGSPAGSPTSTEEGSSTPSGATGSPGSSPSAST

GTGPGASPGTSSTGSPGTSESATPESGPGTSTEPSEGSAPGTSTEPSEGSAP

AM13 31 GTSTEPSEGSAPGSEPATSGSETPGSPAGSPTSTEEGSTSSTAESPGPGTSTPESGSA 18 SPGSTSESPSGTAPGSTSESPSGTAPGTSTPESGSASPGTSTPESGSASPGSEPATSGS

ETPGTSESATPESGPGSPAGSPTSTEEGTSTEPSEGSAPGTSESATPESGPGTSTEPSE

GSAPGTSTEPSEGSAPGSPAGSPTSTEEGTSTEPSEGSAPGTSTEPSEGSAPGTSESA

TPESGPGTSESATPESGPGTSTEPSEGSAPGTSTEPSEGSAPGTSESATPESGPGTST

EPSEGSAPGSEPATSGSETPGSPAGSPTSTEEGSSTPSGATGSPGTPGSGTASSSPGS

STPSGATGSPGTSTEPSEGSAPGTSTEPSEGSAPGSEPATSGSETPGSPAGSPTSTEE

GSPAGSPTSTEEGTSTEPSEGSAPGPEPTGPAPSGGSEPATSGSETPGTSESATPESG

PGSPAGSPTSTEEGTSESATPESGPGSPAGSPTSTEEGSPAGSPTSTEEGTSESATPE

SGPGSPAGSPTSTEEGSPAGSPTSTEEGSTSSTAESPGPGSTSESPSGTAPGTSPSGES

STAPGSTSESPSGTAPGSTSESPSGTAPGTSPSGESSTAPGTSTEPSEGSAPGTSESA

TPESGPGTSESATPESGPGSEPATSGSETPGTSESATPESGPGTSESATPESGPGTST

EPSEGSAPGTSESATPESGPGTSTEPSEGSAPGTSPSGESSTAPGTSPSGESSTAPGT

SPSGESSTAPGTSTEPSEGSAPGSPAGSPTSTEEGTSTEPSEGSAPGSSPSASTGTGP

GSSTPSGATGSPGSSTPSGATGSPGSSTPSGATGSPGSSTPSGATGSPGASPGTSSTG

SPGASASGAPSTGGTSPSGESSTAPGSTSSTAESPGPGTSPSGESSTAPGTSESATPE

SGPGTSTEPSEGSAPGTSTEPSEGSAPGSSPSASTGTGPGSSTPSGATGSPGASPGTS

STGSPGTSTPESGSASPGTSPSGESSTAPGTSPSGESSTAPGTSESATPESGPGSEPAT

SGSETPGTSTEPSEGSAPGSTSESPSGTAPGSTSESPSGTAPGTSTPESGSASPGSPA

GSPTSTEEGTSESATPESGPGTSTEPSEGSAPGSPAGSPTSTEEGTSESATPESGPGS

EPATSGSETPGSSTPSGATGSPGASPGTSSTGSPGSSTPSGATGSPGSTSESPSGTAP

GTSPSGESSTAPGSTSSTAESPGPGSSTPSGATGSPGASPGTSSTGSPGTPGSGTASS

SPGSPAGSPTSTEEGSPAGSPTSTEEGTSTEPSEGSAP

BC 32 GTSTEPSEPGSAGTSTEPSEPGSAGSEPATSGTEPSGSGASEPTSTEPGSEPATSGTE 864 PSGSEPATSGTEPSGSEPATSGTEPSGSGASEPTSTEPGTSTEPSEPGSAGSEPATSG

TEPSGTSTEPSEPGSAGSEPATSGTEPSGSEPATSGTEPSGTSTEPSEPGSAGTSTEPS

EPGSAGSEPATSGTEPSGSEPATSGTEPSGTSEPSTSEPGAGSGASEPTSTEPGTSEP

STSEPGAGSEPATSGTEPSGSEPATSGTEPSGTSTEPSEPGSAGTSTEPSEPGSAGSG

ASEPTSTEPGSEPATSGTEPSGSEPATSGTEPSGSEPATSGTEPSGSEPATSGTEPSG

TSTEPSEPGSAGSEPATSGTEPSGSGASEPTSTEPGTSTEPSEPGSAGSEPATSGTEP

SGSGASEPTSTEPGTSTEPSEPGSAGSGASEPTSTEPGSEPATSGTEPSGSGASEPTS

TEPGSEPATSGTEPSGSGASEPTSTEPGTSTEPSEPGSAGSEPATSGTEPSGSGASEP

TSTEPGTSTEPSEPGSAGSEPATSGTEPSGTSTEPSEPGSAGSEPATSGTEPSGTSTEP

SEPGSAGTSTEPSEPGSAGTSTEPSEPGSAGTSTEPSEPGSAGTSTEPSEPGSAGTST

EPSEPGSAGTSEPSTSEPGAGSGASEPTSTEPGTSTEPSEPGSAGTSTEPSEPGSAGT

STEPSEPGSAGSEPATSGTEPSGSGASEPTSTEPGSEPATSGTEPSGSEPATSGTEPS

GSEPATSGTEPSGSEPATSGTEPSGTSEPSTSEPGAGSEPATSGTEPSGSGASEPTST

EPGTSTEPSEPGSAGSEPATSGTEPSGSGASEPTSTEPGTSTEPSEPGSA

BD864 33 GSETATSGSETAGTSESATSESGAGSTAGSETSTEAGTSESATSESGAGSETATSGS

ETAGSETATSGSETAGTSTEASEGSASGTSTEASEGSASGTSESATSESGAGSETAT SGSETAGTSTEASEGSASGSTAGSETSTEAGTSESATSESGAGTSESATSESGAGSE TATSGSETAGTSESATSESGAGTSTEASEGSASGSETATSGSETAGSETATSGSETA GTSTEASEGSASGSTAGSETSTEAGTSESATSESGAGTSTEASEGSASGSETATSGS ETAGSTAGSETSTEAGSTAGSETSTEAGSETATSGSETAGTSESATSESGAGTSESA TSESGAGSETATSGSETAGTSESATSESGAGTSESATSESGAGSETATSGSETAGSE TATSGSETAGTSTEASEGSASGSTAGSETSTEAGSETATSGSETAGTSESATSESGA GSTAGSETSTEAGSTAGSETSTEAGSTAGSETSTEAGTSTEASEGSASGSTAGSETS SEQ

XTEN ID Amino Acid Sequence

Name

NO:

TEAGSTAGSETSTEAGTSTEASEGSASGSTAGSETSTEAGSETATSGSETAGTSTEA

SEGSASGTSESATSESGAGSETATSGSETAGTSESATSESGAGTSESATSESGAGSE

TATSGSETAGTSESATSESGAGSETATSGSETAGTSTEASEGSASGTSTEASEGSAS

GSTAGSETSTEAGSTAGSETSTEAGSETATSGSETAGTSESATSESGAGTSESATSE

SGAGSETATSGSETAGSETATSGSETAGSETATSGSETAGTSTEASEGSASGTSESA

TSESGAGSETATSGSETAGSETATSGSETAGTSESATSESGAGTSESATSESGAGSE

TATSGSETA

AE911 34 AEPAGSPTSTEEGTPGSGTASSSPGSSTPSGATGSPGASPGTSSTGSPGSPAGSPTST

EEGTSESATPESGPGTSTEPSEGSAPGSPAGSPTSTEEGTSTEPSEGSAPGTSTEPSE

GSAPGTSESATPESGPGSEPATSGSETPGSEPATSGSETPGSPAGSPTSTEEGTSESA

TPESGPGTSTEPSEGSAPGTSTEPSEGSAPGSPAGSPTSTEEGTSTEPSEGSAPGTST

EPSEGSAPGTSESATPESGPGTSTEPSEGSAPGTSESATPESGPGSEPATSGSETPGT

STEPSEGSAPGTSTEPSEGSAPGTSESATPESGPGTSESATPESGPGSPAGSPTSTEE

GTSESATPESGPGSEPATSGSETPGTSESATPESGPGTSTEPSEGSAPGTSTEPSEGS

APGTSTEPSEGSAPGTSTEPSEGSAPGTSTEPSEGSAPGTSTEPSEGSAPGSPAGSPT

STEEGTSTEPSEGSAPGTSESATPESGPGSEPATSGSETPGTSESATPESGPGSEPAT

SGSETPGTSESATPESGPGTSTEPSEGSAPGTSESATPESGPGSPAGSPTSTEEGSPA

GSPTSTEEGSPAGSPTSTEEGTSESATPESGPGTSTEPSEGSAPGTSESATPESGPGS

EPATSGSETPGTSESATPESGPGSEPATSGSETPGTSESATPESGPGTSTEPSEGSAP

GSPAGSPTSTEEGTSESATPESGPGSEPATSGSETPGTSESATPESGPGSPAGSPTST

EEGSPAGSPTSTEEGTSTEPSEGSAPGTSESATPESGPGTSESATPESGPGTSESATP

ESGPGSEPATSGSETPGSEPATSGSETPGSPAGSPTSTEEGTSTEPSEGSAPGTSTEP

SEGSAPGSEPATSGSETPGTSESATPESGPGTSTEPSEGSAP

AE146 35 GGTSESATPESGPGTSTEPSEGSAPGTSTEPSEGSAPGTSESATPESGPGTSTEPSEG

SAPGTSTEPSEGSAPGTSESATPESGPGTSTEPSEGSAPGTSTEPSEGSAPGTSTEPS EGSAPGSPAGSPTSTEEGTSTEPSEGSAPG

AE48. 36 AEPAGSPTSTEEGTPGSGTASSSPGSSTPSGATGSPGASPGTSSTGS

1

AM48. 37 AEPAGSPTSTEEGASPGTSSTGSPGSSTPSGATGSPGSSTPSGATGS

1

AE912 38 AEPAGSPTSTEEGTPGSGTASSSPGSSTPSGATGSPGASPGTSSTGSPGSPAGSPTST .1 EEGTSESATPESGPGTSTEPSEGSAPGSPAGSPTSTEEGTSTEPSEGSAPGTSTEPSE

GSAPGTSESATPESGPGSEPATSGSETPGSEPATSGSETPGSPAGSPTSTEEGTSESA

TPESGPGTSTEPSEGSAPGTSTEPSEGSAPGSPAGSPTSTEEGTSTEPSEGSAPGTST

EPSEGSAPGTSESATPESGPGTSTEPSEGSAPGTSESATPESGPGSEPATSGSETPGT

STEPSEGSAPGTSTEPSEGSAPGTSESATPESGPGTSESATPESGPGSPAGSPTSTEE

GTSESATPESGPGSEPATSGSETPGTSESATPESGPGTSTEPSEGSAPGTSTEPSEGS

APGTSTEPSEGSAPGTSTEPSEGSAPGTSTEPSEGSAPGTSTEPSEGSAPGSPAGSPT

STEEGTSTEPSEGSAPGTSESATPESGPGSEPATSGSETPGTSESATPESGPGSEPAT

SGSETPGTSESATPESGPGTSTEPSEGSAPGTSESATPESGPGSPAGSPTSTEEGSPA

GSPTSTEEGSPAGSPTSTEEGTSESATPESGPGTSTEPSEGSAPGTSESATPESGPGS

EPATSGSETPGTSESATPESGPGSEPATSGSETPGTSESATPESGPGTSTEPSEGSAP

GSPAGSPTSTEEGTSESATPESGPGSEPATSGSETPGTSESATPESGPGSPAGSPTST

EEGSPAGSPTSTEEGTSTEPSEGSAPGTSESATPESGPGTSESATPESGPGTSESATP

ESGPGSEPATSGSETPGSEPATSGSETPGSPAGSPTSTEEGTSTEPSEGSAPGTSTEP

SEGSAPGSEPATSGSETPGTSESATPESGPGTSTEPSEGSAP

AE912 39 AEPAGSPTSTEEGTPGSGTASSSPGSSTPSGATGSPGASPGTSSTGSPGSPAGSPTST .2 EEGTSESATPESGPGTSTEPSEGSAPGSPAGSPTSTEEGTSTEPSEGSAPGTSTEPSE

GSAPGTSESATPESGPGSEPATSGSETPGSEPATSGSETPGSPAGSPTSTEEGTSESA

TPESGPGTSTEPSEGSAPGTSTEPSEGSAPGSPAGSPTSTEEGTSTEPSEGSAPGTST

EPSEGSAPGTSESATPESGPGTSTEPSEGSAPGTSESATPESGPGSEPATSGSETPGT

STEPSEGSAPGTSTEPSEGSAPGTSESATPESGPGTSESATPESGPGSPAGSPTSTEE

GTSESATPESGPGSEPATSGSETPGTSESATPESGPGTSTEPSEGSAPGTSTEPSEGS

APGTSTEPSEGSAPGTSTEPSEGSAPGTSTEPSEGSAPGTSTEPSEGSAPGSPAGSPT

STEEGTSTEPSEGSAPGTSESATPESGPGSEPATSGSETPGTSESATPESGPGSEPAT

SGSETPGTSESATPESGPGTSTEPSEGSAPGTSESATPESGPGSPAGSPTSTEEGSPA SEQ

XTEN ID Amino Acid Sequence

Name

NO:

GSPTSTEEGSPAGSPTSTEEGTSESATPESGPGTSTEPSEGSAPGTSESATPESGPGS EPATSGSETPGTSESATPESGPGSEPATSGSETPGTSESATPESGPGTSTEPSEGSAP GSPAGSPTSTEEGTSESATPESGPGSEPATSGSETPGTSESATPESGPGSPAGSPTST EEGSPAGSPTSTEEGTSTEPSEGSAPGTSESATPESGPGTSESATPESGPGTSESATP ESGPGSEPATSGSETPGSEPATSGSETPGSPAGSPTSTEEGTSTEPSEGSAPGTSTEP SEGSAPGSEPATSGSETPGTSESATPESGPGTSTEPSEGSAPG

AE146 40 TSESATPESGPGTSTEPSEGSAPGTSTEPSEGSAPGTSESATPESGPGTSTEPSEGSA .1 PGTSTEPSEGSAPGTSESATPESGPGTSTEPSEGSAPGTSTEPSEGSAPGTSTEPSEG

SAPGSPAGSPTSTEEGTSTEPSEGSAPG

AM87 41 GTSTEPSEGSAPGSEPATSGSETPGSPAGSPTSTEEGSTSSTAESPGPGTSTPESGSA 5.1 SPGSTSESPSGTAPGSTSESPSGTAPGTSTPESGSASPGTSTPESGSASPGSEPATSGS

ETPGTSESATPESGPGSPAGSPTSTEEGTSTEPSEGSAPGTSESATPESGPGTSTEPSE

GSAPGTSTEPSEGSAPGSPAGSPTSTEEGTSTEPSEGSAPGTSTEPSEGSAPGTSESA

TPESGPGTSESATPESGPGTSTEPSEGSAPGTSTEPSEGSAPGTSESATPESGPGTST

EPSEGSAPGSEPPTSTEEGSSTPSGATGSPGTPGSGTASSSPGSSTPSGATATSGSET

PGSPAGSGSPGTSTEPSEGSAPGTSTEPSEGSAPGSEPATSGSETPGSPAGSPTSTEE

GSPAGSPTSTEEGTSTEPSEGSAPGASASGAPSTGGTSESATPESGPGSPAGSPTSTE

EGSPAGSPTSTEEGSTSSTAESPGPGSTSESPSGTAPGTSPSGESSTAPGTPGSGTAS

SSPGSSTPSGATGSPGSSPSASTGTGPGSEPATSGSETPGTSESATPESGPGSEPATS

GSETPGSTSSTAESPGPGSTSSTAESPGPGTSPSGESSTAPGSEPATSGSETPGSEPA

TSGSETPGTSTEPSEGSAPGSTSSTAESPGPGTSTPESGSASPGSTSESPSGTAPGTST

EPSEGSAPGTSTEPSEGSAPGTSTEPSEGSAPGSSTPSGATGSPGSSPSASTGTGPGA

SPGTSSTGSPGSEPATSGSETPGTSESATPESGPGSPAGSPTSTEEGSSTPSGATGSP

GSSPSASTGTGPGASPGTSSTGSPGTSESATPESGPGTSTEPSEGSAPGTSTEPSEGS

AP

AM87 42 GTSTEPSEGSAPGSEPATSGSETPGSPAGSPTSTEEGSTSSTAESPGPGTSTPESGSA 5.2 SPGSTSESPSGTAPGSTSESPSGTAPGTSTPESGSASPGTSTPESGSASPGSEPATSGS

ETPGTSESATPESGPGSPAGSPTSTEEGTSTEPSEGSAPGTSESATPESGPGTSTEPSE

GSAPGTSTEPSEGSAPGSPAGSPTSTEEGTSTEPSEGSAPGTSTEPSEGSAPGTSESA

TPESGPGTSESATPESGPGTSTEPSEGSAPGTSTEPSEGSAPGTSESATPESGPGTST

EPSEGSAPGSEPPTSTEEGSSTPSGATGSPGTPGSGTASSSPGSSTPSGATATSGSET

PGSPAGSGSPGTSTEPSEGSAPGTSTEPSEGSAPGSEPATSGSETPGSPAGSPTSTEE

GSPAGSPTSTEEGTSTEPSEGSAPGASASGAPSTGGTSESATPESGPGSPAGSPTSTE

EGSPAGSPTSTEEGSTSSTAESPGPGSTSESPSGTAPGTSPSGESSTAPGTPGSGTAS

SSPGSSTPSGATGSPGSSPSASTGTGPGSEPATSGSETPGTSESATPESGPGSEPATS

GSETPGSTSSTAESPGPGSTSSTAESPGPGTSPSGESSTAPGSEPATSGSETPGSEPA

TSGSETPGTSTEPSEGSAPGSTSSTAESPGPGTSTPESGSASPGSTSESPSGTAPGTST

EPSEGSAPGTSTEPSEGSAPGTSTEPSEGSAPGSSTPSGATGSPGSSPSASTGTGPGA

SPGTSSTGSPGSEPATSGSETPGTSESATPESGPGSPAGSPTSTEEGSSTPSGATGSP

GSSPSASTGTGPGASPGTSSTGSPGTSESATPESGPGTSTEPSEGSAPGTSTEPSEGS

APG

In those embodiments wherein the XTEN component of the hGH-XTEN fusion protein has less than 100% of its amino acids consisting of 4, 5, or 6 types of amino acid selected from glycine (G), alanine (A), serine (S), threonine (T), glutamate (E) and proline (P), or less than 100% of the sequence consisting of the XTEN sequences of Table 2, the other amino acid residues of the XTEN are selected from any of the other 14 natural L-amino acids, but are preferentially selected from hydrophilic amino acids such that the XTEN sequence contains at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least about 99% hydrophilic amino acids. The XTEN amino acids that are not glycine (G), alanine (A), serine (S), threonine (T), glutamate (E) and proline (P) are either interspersed throughout the XTEN sequence, are located within or between the sequence motifs, or are concentrated in one or more short stretches of the XTEN sequence, e.g., to create a linker between the XTEN and the hGH components. In such cases where the XTEN component of the hGH-XTEN comprises amino acids other than glycine (G), alanine (A), serine (S), threonine (T), glutamate (E) and proline (P), it is preferred that less than about 2% or less than about 1% of the amino acids be hydrophobic residues such that the resulting sequences generally lack secondary structure, e.g., not having more than 2% alpha helices or 2% beta- sheets, as determined by the methods disclosed herein. Hydrophobic residues that are less favored in construction of XTEN include tryptophan, phenylalanine, tyrosine, leucine, isoleucine, valine, and methionine. Additionally, one can design the XTEN sequences to contain less than 5% or less than 4% or less than 3% or less than 2% or less than 1% or none of the following amino acids: cysteine (to avoid disulfide formation and oxidation), methionine (to avoid oxidation), asparagine and glutamine (to avoid deamidation). Thus, in some embodiments, the XTEN component of the hGH-XTEN fusion protein comprising other amino acids in addition to glycine (G), alanine (A), serine (S), threonine (T), glutamate (E) and proline (P) have a sequence with less than 5% of the residues contributing to alpha- helices and beta-sheets as measured by the Chou-Fasman algorithm and have at least 90%, or at least about 95% or more random coil formation as measured by the GOR algorithm.

In some embodiments, the hGH-XTEN of the formulation comprises i) one or more

XTEN sequence wherein at least about 80% the XTEN sequence consist of non-overlapping sequence motifs, wherein each sequence motif has 9 to 14 amino acid residues, wherein each motif consist of 4 to 6 types of amino acids selected from glycine (G), alanine (A), serine (S), threonine (T), glutamate (E) and praline (P), the sequence of any two contiguous amino acid residues in any one sequence motif does not occur more than twice in the sequence motif and the XTEN sequence contains no three contiguous amino acids that are identical unless the amino acids are serine residues, ii) where at least one XTEN is linked to either the N- and C- termini of at least one hGH, and wherein when more than one XTEN is used in a hGH-XTEN composition an XTEN can be linked to both the N- and C-termini of at least one hGH, iii) the length of the N-terminal XTEN sequence ranges between about 500 amino acids to about 1320 amino acids and the length of the C-terminal XTEN ranges between about 40 amino acids to about 300 amino acids, iv) when the XTEN component of the hGH-XTEN fusion protein has less than 100% of its amino acids consisting of 4, 5, or 6 types of amino acid selected from glycine (G), alanine (A), serine (S), threonine (T), glutamate (E) and proline (P), the other amino acid residues of the XTEN are selected from any of the other 14 natural L-amino acids, but are preferentially selected from hydrophilic amino acids such that the XTEN sequence contains at least about 90% to at least about 99% hydrophilic amino acids, v) the XTEN amino acids that are not glycine (G), alanine (A), serine (S), threonine (T), glutamate (E) and proline (P) are either interspersed throughout the XTEN sequence, are located within or between the sequence motifs, or are concentrated in one or more short stretches of the XTEN sequence, e.g., to create a linker between the XTEN and the hGH components, vi) less than about 2% or less than about 1% of the amino acids are hydrophobic residues such that the resulting sequences generally lack secondary structure, e.g., not having more than 2% alpha helices or 2% beta-sheets, and vii) the XTEN sequences are designed to contain less than 5% or less than 4% or less than 3% or less than 2% or less than 1% or none of the following amino acids: cysteine, methionine, asparagine and glutamine. In some embodiments, the XTEN sequence exhibits at least about 80% to about 100% sequence identity to an XTEN selected from Table 2.

VII) . hGH-XTEN STRUCTURAL CONFIGURATIONS AND PROPERTIES

The human growth hormone (hGH) of the subject formulations are not limited to native, full-length polypeptides, but also include recombinant versions as well as biologically and/or pharmacologically active variants or fragments thereof. For example, it will be appreciated that various amino acid deletions, insertions and substitutions can be made in the hGH to create variants without departing from the spirit of the invention with respect to the biological activity or pharmacologic properties of the hGH. For a more detailed discussion of possible structural configurations and properties of hGH-XTEN sequences in the subject formulations, including amino acid substitutions and fusion protein configurations, see

W013/184216, WO/2014/164568, U.S. Patent Nos. 8,492,530 and 8,703,717, each of which is incorporated herein by reference in its entirety.

VIII) . USES OF THE FORMULATIONS OF THE PRESENT INVENTION

In another aspect, the invention provides a method for achieving a beneficial therapeutic effect in a disease, disorder or condition mediated by hGH by therapeutic treatment with a storage stable aqueous liquid formulation comprising an hGH-XTEN fusion protein. The present invention addresses disadvantages and/or limitations of prior aqueous liquid hGH formulations, which are unstable in aqueous solution over time, have a relatively short terminal half-life and/or a narrow therapeutic window. The fact that growth hormone has a short half-life, necessitates frequent dosing in order to achieve clinical benefit, which results in difficulties in the management of such patients. Methods suitable for treating a hGH-related condition in a subject using the hGH-XTEN formulation of the present invention are known in the art and are described in, for example, in U.S. Patent No. 8,703,717, in WO/2010/144502, in WO/2013/184216 and WO/2014/164568. All embodiments of the methods of treatment described or incorporated by reference herein apply to all method and use claims recited in this application.

In one embodiment, the invention provides a method for achieving a beneficial effect in a subject with a growth hormone-related disease, disorder or condition comprising the step of administering to the subject a therapeutically- or prophylactically-effective amount of a hGH-XTEN formulation, wherein said administration results in the improvement of one or more biochemical or physiological parameters or clinical endpoints associated with a growth hormone-related disease, disorder or condition. The effective amount produces a beneficial effect in helping to treat (e.g., cure or reduce the severity) or prevent (e.g., reduce the likelihood of onset or severity) a growth hormone-related disease, disorder or condition. In some cases, the method for achieving a beneficial effect includes administering a

therapeutically effective amount of a hGH-XTEN fusion protein composition to treat a subject with a growth hormone-related disease, disorder, or condition, including, but not limited to, congenital or acquired GH deficiency in adults and children, Turner's Syndrome, Prader-Willi Syndrome, chronic renal failure, intrauterine growth retardation, idiopathic short stature, AIDS wasting, obesity, multiple sclerosis, aging, fibromyalgia, Crohn's disease, ulcerative colitis, muscular dystrophy, low muscle mass (e.g. bodybuilding), low bone density, or any other indication for which GH can be utilized (but for which endogenous growth hormone levels in a subject are not necessarily deficient).

In a preferred embodiment, the therapeutic effect is a measured parameter selected from, for example, IGF-1 concentrations, IGFBP3 concentration, height velocity, lean body mass, total body fat, trunk fat, response to insulin challenge, rate of division of chondrocytes, chondrocyte numbers, bone density, bone growth, and increase in epiphyseal plate width.

Determination of a therapeutically effective amount is well within the capability of those skilled in the art. In a preferred embodiment, therapeutically effective amounts result in a sustained beneficial effect on any clinical sign or symptom, aspect, measured parameter or characteristic of a metabolic disease state or condition, including, but not limited to, those described herein. In one embodiment, the parameters include but are not limited to mean (SD) height standard deviation score (HT-SDS), changes in height velocity, IGF-I concentration, ratio of IGF-I/IGFBP-3, IGFBP3 concentration, change in weight, lean body mass, change in body mass index, total body fat (adipose fat/tissue), trunk fat, response to insulin challenge, rate of division of chondrocytes, chondrocyte numbers, bone density, bone age, bone growth, bone turnover, increase in epiphyseal plate width, reduction in cholesterol, reduction in triglycerides, and reduction in LDL. In another embodiment of the method, the administration to a human pediatric patient of successive or consecutive doses of a therapeutically effective amount of the hGH-XTEN results in a beneficial effect in two or more of the parameters including, but not limited to mean (SD) height standard deviation score (HT-SDS), changes in height velocity, IGF-I concentration, ratio of IGF-I/IGFBP-3, IGFBP3 concentration, change in weight, lean body mass, change in body mass index, total body fat (adipose fat/tissue), trunk fat, response to insulin challenge, rate of division of chondrocytes, chondrocyte numbers, bone density, bone age, bone growth, bone turnover, increase in epiphyseal plate width, reduction in cholesterol, reduction in triglycerides, and reduction in LDL.

In another embodiment, the invention provides a method of stimulating IGF- 1 production in individuals with GH deficiency. The method comprises the step of

administering therapeutically effective amount of an hGH-XTEN to a subject that results in the increased blood levels and/or duration in increased blood levels of IGF-I compared to a subject receiving a GH not linked to an XTEN and administered at a comparable dose. In another embodiment, the invention provides a method of stimulating the division and numbers of chrondrocytes. In another embodiment, the invention provides a method comprising the step of administering therapeutically effective amount of hGH-XTEN that results in increased bone growth as measured by increase in epiphyseal plate width compared to a subject receiving a GH not linked to an XTEN and administered at a comparable dose.

As a result of the enhanced pharmacokinetic parameters of the hGH-XTEN formulation, the hGH can be administered using longer intervals between doses compared to the corresponding GH not linked to XTEN to prevent, treat, alleviate, reverse or ameliorate symptoms or clinical abnormalities of the growth hormone- related disease, disorder or condition or prolong the survival of the subject being treated. The methods of the invention includes administration of consecutive doses of a therapeutically effective amount of the hGH-XTEN for a period of time sufficient to achieve and/or maintain the desired parameter or clinical effect, and such consecutive doses of a therapeutically effective amount establishes the therapeutically effective dose regimen for the hGH-XTEN; i.e., the schedule for consecutively administered doses of the fusion protein composition, wherein the doses are given in therapeutically effective amounts to result in a sustained beneficial effect on any clinical sign or symptom, aspect, measured parameter or characteristic of a metabolic disease state or condition, including, but not limited to, those described herein. In one embodiment, the method comprises administering a therapeutically- effective amount of a pharmaceutical composition comprising a hGH-XTEN fusion protein composition comprising a GH linked to an XTEN sequence(s) and at least one

pharmaceutically acceptable carrier to a subject in need thereof that results in greater improvement in at least one parameter, physiologic condition, or clinical outcome mediated by the GH component(s) (non- limiting examples of which are described above) compared to the effect mediated by administration of a pharmaceutical composition comprising a GH not linked to XTEN and administered at a comparable dose. In one embodiment, the

pharmaceutical composition is administered at a therapeutically effective dose. In another embodiment, the pharmaceutical composition is administered using multiple consecutive doses using a therapeutically effective dose regimen (as defined herein) for the length of the dosing period.

A therapeutically effective amount of the hGH-XTEN varies according to factors such as the disease state, age, sex, and weight of the individual, and the ability of the hGH to elicit a desired response in the individual. A therapeutically effective amount is also one in which any toxic or detrimental effects of the hGH-XTEN are outweighed by the therapeutically beneficial effects. A prophylactically effective amount refers to an amount of hGH-XTEN required for the period of time necessary to achieve the desired prophylactic result.

For the inventive methods, longer acting hGH-XTEN compositions are preferred, so as to improve patient convenience, to increase the interval between doses and to reduce the amount of drug required to achieve a sustained effect. In one embodiment, a method of treatment comprises administration of a therapeutically effective dose of a hGH-XTEN to a subject in need thereof that results in a gain in time spent within a therapeutic window established for the fusion protein of the composition compared to the corresponding GH component(s) not linked to the fusion protein and administered at a comparable dose to a subject. In some cases, the gain in time spent within the therapeutic window is at least about three-fold to at least about 100-fold longer, compared to the corresponding GH component not linked to the fusion protein and administered at a comparable dose to a subject.

The methods further provide that administration of multiple consecutive doses of a hGH-XTEN administered using a therapeutically effective dose regimen to a subject in need thereof results in a gain in time between consecutive C_max peaks and/or C_min troughs for blood levels of the fusion protein compared to the corresponding GH not linked to the fusion protein and administered using a dose regimen established for that GH. In the foregoing embodiment, the gain in time spent between consecutive C_max peaks and/or Cmin troughs is at least about three-fold to at least about 100-fold longer, compared to the corresponding GH component not linked to the fusion protein and administered using a dose regimen established for that GH. In the embodiments hereinabove described in this paragraph the administration of the fusion protein results in an improvement in at least one of the parameters (disclosed herein as being useful for assessing the subject diseases, conditions or disorders) using a lower unit dose in moles of fusion protein compared to the corresponding GH component not linked to the fusion protein and administered at a comparable unit dose or dose regimen to a subject.

The method of treatment comprises administration of a hGH-XTEN formulation using a therapeutically effective dose regimen to effect improvements in one or more parameters associated with growth hormone diseases, disorders or conditions. In some embodiments, administration of the hGH-XTEN formulation to a subject results in an improvement in one or more of the biochemical, physiologic, or clinical parameters that is of greater magnitude than that of the corresponding GH component not linked to XTEN, determined using the same assay or based on a measured clinical parameter. In other embodiments, administration of the hGH-XTEN formulation to a subject using a therapeutically effective dose regimen results in activity in one or more of the biochemical, physiologic, or clinical parameters that is of longer duration than the activity of one of the single GH components not linked to XTEN, determined using that same assay or based on a measured clinical parameter. In one embodiment of the foregoing, the administration of the hGH-XTEN formulation to a subject using a therapeutically effective dose regimen results in an improvement in peak

concentrations and area under the curve of blood IGF-I levels of at least about 10% to about 100% or more in the subject compared to a comparable dose of GH not linked to XTEN administered to a subject. In another embodiment of the foregoing, the administration of the hGH-XTEN formulation to a subject using a therapeutically effective dose regimen results in increased weight gain in the subject of at least about 10% to about 50% or more compared to a comparable dose regimen of GH not linked to XTEN administered to a subject.

The invention further contemplates that hGH-XTEN used in accordance with the methods provided herein is administered in conjunction with other treatment methods and pharmaceutical compositions useful for treating growth hormone-related diseases, disorders, and conditions, or conditions for which growth hormone is adjunctive therapy; e.g., insulin resistance and poor glycemic control. Such compositions, include for example, DPP-IV inhibitors, insulin, insulin analogues, PPAR gamma agonists, dual-acting PPAR agonists, GLP-1 agonists or analogues, PTPIB inhibitors, SGLT inhibitors, insulin secretagogues, RXR agonists, glycogen synthase kinase-3 inhibitors, insulin sensitizers, immune modulators, beta- 3 adrenergic receptor agonists, pan-PPAR agonists, 11 beta-HSDl inhibitors, biguanides, alpha-glucosidase inhibitors, meglitinides, thiazolidinediones, sulfonylureas and other diabetes medicants known in the art, or anti-hypertensive drugs, calcium channel blockers, and related products. In some embodiments, the administration of a hGH-XTEN formulation permits use of lower dosages of the co-administered pharmaceutical composition to achieve a comparable clinical effect or measured parameter for the disease, disorder or condition in the subject. IX). PHARMACEUTICAL COMPOSITIONS

The present invention provides pharmaceutical compositions comprising hGH-XTEN. In one embodiment, the pharmaceutical composition comprises the hGH-XTEN fusion protein and at least one pharmaceutically acceptable carrier. hGH-XTEN polypeptides of the present invention can be formulated according to known methods to prepare

pharmaceutically useful compositions, whereby the polypeptide is combined in admixture with a pharmaceutically acceptable carrier vehicle, such as aqueous solutions or buffers, pharmaceutically acceptable suspensions and emulsions. Examples of non-aqueous solvents include propyl ethylene glycol, polyethylene glycol and vegetable oils. Therapeutic formulations are prepared for storage by mixing the active ingredient having the desired degree of purity with optional physiologically acceptable carriers, excipients or stabilizers, as described in Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980), in the form of lyophilized formulations or aqueous solutions. In addition, the pharmaceutical compositions can also contain other pharmaceutically active compounds or a plurality of compounds of the invention.

The present pharmaceutical compositions may be administered for therapy by any suitable route established in the art. It will also be appreciated that the preferred route will vary with the condition and age of the recipient, and the disease being treated. In one embodiment, the pharmaceutical composition is administered subcutaneously. In this embodiment, the composition may be supplied as a lyophilized powder to be reconstituted prior to administration. The composition may also be supplied in a liquid form, which can be administered directly to a patient. In one embodiment, the composition is supplied as a liquid in a pre-filled syringe such that a patient can easily self-administer the composition.

The compositions of the invention may also be formulated using a variety of excipients known in the art. Suitable excipients include microcrystalline cellulose (e.g. Avicel® PH-102, Avicel® PH-101), polymethacrylate, poly(ethyl acrylate, methyl methacrylate, trimethylammomoethyl methacrylate chloride) (such as Eudragit® RS-30D), hydroxypropyl methylcellulose (Methocel™ K100M, Premium CR Methocel™ K100M, Methocel™ E5, Opadry®), magnesium stearate, talc, triethyl citrate, aqueous ethylcellulose dispersion (Surelease®), and protamine sulfate. The compositions may also comprise a carrier, which can comprise, for example, solvents, dispersion media, antibacterial and antifungal agents, isotonic and absorption delaying agents. Pharmaceutically acceptable salts may also be used in these compositions, for example, mineral salts such as hydrochlorides, hydrobromides, phosphates, or sulfates, as well as the salts of organic acids such as acetates, proprionates, malonates, or benzoates. The composition may also contain liquids, such as water, saline, glycerol, and ethanol, as well as substances such as wetting agents, emulsifying agents, or pH buffering agents. Liposomes may also be used as a carrier.

For liquid formulations, a desired property is that the formulation be supplied in a form that can pass through a 25, 28, 30, 31, 32 gauge needle for intravenous, intramuscular, intraarticular, or subcutaneous administration.

X). PHARMACEUTICAL KITS

In another aspect, the invention provides a kit to facilitate the use of the hGH-XTEN formulation. The kit comprises the pharmaceutical composition provided herein, a label identifying the pharmaceutical composition, and an instruction for storage, reconstitution and/or administration of the pharmaceutical compositions to a subject. In some embodiments, the kit comprises, preferably: (a) an amount of a hGH-XTEN fusion protein composition sufficient to treat a disease, condition or disorder upon administration to a subject in need thereof; and (b) an amount of a pharmaceutically acceptable carrier; together in a formulation ready for injection or for reconstitution with sterile water, buffer, or dextrose; together with a label identifying the hGH-XTEN drug and storage and handling conditions, and a sheet of the approved indications for the drug, instructions for the reconstitution and/or administration of the hGH-XTEN drug for the use for the prevention and/or treatment of approved indication, appropriate dosage and safety information, and information identifying the lot and expiration of the drug. In another embodiment of the foregoing, the kit can comprise a second container that can carry a suitable diluent for the hGH-XTEN composition, which will provide the user with the appropriate concentration of hGH-XTEN to be delivered to the subject.

EXPERIMENTAL EXAMPLES

Example I - Reference Assay Methods

Reverse-phase high-performance liquid chromatography (RP-HPLC) was run on the Agilent® 1100 HPLC system using a Kinetex® reversed phase C18 column (2.6 μπι, 150 x 4.6mm, Phenomenex®, P/N 00F-4462-E0) at a column temperature of 40° C with a flow rate of 1 ml/min. Mobile phase A was 0.1% TFA in water and mobile phase B was 0.1% TFA in acetonitrile. Elution was performed using a 15 minute gradient from 60-90%) mobile phase B beginning at 95 minutes. The total run time was 105 minutes. The column load was 40 μg of protein. Detection was at 214 nm.

Size exclusion high-performance liquid chromatography (SE-HPLC) was run on the Agilent® 1100 HPLC system using a Phenomenex® Bio-Sep-SEC-S4000 column

(7.8x600mm, part # 00K-2147-KO) at ambient column temperature with a Phenomenex®

SecurityGuard with GFC 4000 cartridge (Part # KJO-4282) guard column with a flow rate of 0.5 ml/min. The mobile phase was 20mM Tris, 300mM NaCl at pH 7.5. The total run time was 75 minutes. The column load was 25 μg of protein. Detection was at 215 nm.

Strong anion exchange high-performance liquid chromatography (SAX-HPLC) was run on the Agilent 1100 HPLC system using a ProSwift® SAX-1 S column (4.6 x 50 mm, Thermo®, P/N 64293) at a column temperature of 25° C with a flow rate of 1 ml/min.

Mobile phase A was 20mM Tris at pH 8.0 and mobile phase B was Mobile A + 1.0 M NaCl. Elution was performed using a 28 minute gradient from 5 (at 4 min), 50 (at 25 min), 100 (at 25.01 min), 100 (at 30 min), 5 (at 30.01 min), and 5% (at 32 min) mobile phase B beginning at 4 minutes. The total run time was 32 minutes. The column load was 25 μg of protein. Detection was at 215 nm. Example II - Solubility of the Aqueous Formulation

VRS-317 contains rhGH and two XTEN domains consisting of hydrophilic amino acids and a large number of glutamate residues. The high glutamate content of VRS-317 results in an isoelectric point (pi) of approximately 3.6 compared to the rhGH pi of 5.2. This lower pi allows for the aqueous formulation of VRS-317 at lower pH than rhGH.

The aqueous formulation prepared according to the present invention containing 54 mg/ml of VRS-317 (SEQ ID NO: 1) in 20 mM L-histidine and 154 mM NaCl at pH 5.5 (the reference standard), was concentrated in a step-wise manner up to 200 mg/mL via centrifugal filtration (3 OK MWCO PES) to assess the osmolality and solubility of formulations containing various concentrations of VRS-317. Accordingly, VRS-317 was suspended at 125 mg/ml, 150 mg/ml, 175 mg/ml, and 200 mg/ml. The osmolality of these solutions is shown in Table 3. Compared to a reference standard, the measured concentration of VRS-317 suspended at 125 mg/ml, 150 mg/ml, 175 mg/ml, and 200 mg/ml was 125.0±1.0 , 154.3±4.0, 170.7±3.1, and 208.3±15.7, respectively (n=3, ± Std Dev). VRS-317 was found to be soluble in the aqueous formulation at the concentrations tested. All samples remained clear and free of particulates.

Table 3

Example III - Stability of VRS-317 in the Aqueous Formulation

Hydrolysis, Aggregation, Oxidation, and Deamidation

To assess the suitability of several possible formulations, a formulation screen, which included accelerated stability at 40°C, was performed on batches of VRS-317. The primary mode of degradation that was observed under accelerated stability conditions of VRS-317 formulated in 20 mM L-histidine, 154 mM sodium chloride, pH 5.5 at 40°C was the hydrolysis, or truncation, of the XTEN polypeptide. The truncated products are resolved by anion exchange HPLC as shown in FIG. 2. An increase in the peak area of the leading peak (RRT < main product peak) was observed over a period of four weeks. Several buffers and excipients were evaluated and the results are summarized in Table 4.

These results suggested that VRS-317 formulated according to the present invention was less truncated at higher pH (up to 6.0) in 154 mM sodium chloride as compared to other formulations tested.

Table 4. Summary of VRS-317 Accelerated Stability (40°C) Data. Anion exchange analysis of VRS-317 was performed as described.

2 82.8

4 66.2

0 90.4

1 82.7

4% mannitol

2 79.6

4 59.7

0 90.5

1 81.6

8% sucrose

2 65.2

4 43.8

0 90.9

154 mM sodium 1 85.1

20 mM L-histidine, pH 6.0

chloride 2 76.7

4 68.3

0 90.7

1 85.1

4% mannitol

2 78.4

4 55.3

SE-HPLC and SAX-HPLC analyses were performed on the aqueous formulation containing VRS-317 suspended at 5.0 mg/ml of VRS-317 in 20 mM L-histidine and 154 mM NaCl at pH 5.5 (the reference standard), concentrated in a step-wise manner to 50.0, 125.0, 150.0, 175.0, or 200 mg/mL via centrifugal filtration (30K MWCO PES). No significant change in percent main peak was detected for any of the concentrations tested based on the chromatograms obtained from SAX-HPLC (FIG. 3) and SE-HPLC (FIG. 4) experiments. These SE-HPLC and SAX-HPLC results indicate, respectively, that VRS-317 is not degraded or aggregated in the aqueous liquid formulation according to the present invention. To test whether injection of the formulation through a needle would cause aggregation, the samples were also loaded into 1 ml Luer-lok® tip syringes (BD lot #3234079), passed through 0.5" inch 26 gauge, 27 gauge, or 30 gauge needles, and assayed via SE-HPLC. SE-HPLC results for the samples passed through the 30 gauge needle are shown in FIG. 5, and similar results were obtained for the other wider-diameters tested (data not shown). No aggregation of VRS-317 was detected after the formulation according to the present invention was passed through the various needles.

To demonstrate the ability to detect degradation of VRS-317, oxidation or

deamidation of VRS-317 was induced with hydrogen peroxide or basic pH, respectively. The reverse phase HPLC analysis of tryptic fragments of control VRS-317 (black) or VRS-317 treated with hydrogen peroxide for 72 hours (gray) is shown in FIG. 6. The predicted tryptic fragments for VRS-317 (shown with the N-terminal methionine encoded by the start codon) are displayed in Table 5. Oxidation is observed at two methionine residues, Met928

(equivalent to Metl4, rhGH sequence) and Metl039 (equivalent to Metl25, rhGH sequence). Incubation of VRS-317 with 0.1% hydrogen peroxide for 72 hours at room temperature resulted in a complete loss of the HPLC peaks corresponding to the Met928 and Metl039. Two additional peaks, likely the methionine sulfoxide and sulfone degradation products of Metl039, were observed (FIG. 6, inset B). Similar peaks were not detected for Met928; however, any degradation products may co-elute with the intense peaks that elute a few minutes prior to the native Met928 peptide (FIG. 6, inset A).

To induce deamidation, VRS-317 was subjected to elevated pH (-10) and temperature (40°C) for one week. The reverse phase HPLC analysis of tryptic fragments of control VRS- 317 (black) or VRS-317 stressed with elevated pH and temperatures (gray) is shown in FIG. 7. A significant loss of peak area at 36 minutes was observed (FIG. 7). This peak

corresponds to the tryptic fragment containing Asnl063 and Asnl066 (equivalent to Asnl49 and Asnl52, respectively, in the rhGH sequence), which is prone to deamidation. One or more peaks, with retention times greater than 36 minutes, showed increased peak area in the stressed sample (FIG. 7, inset). Synthetic peptides corresponding to the native and monodesamido forms (Asnl063→Asp) of the tryptic fragment can be resolved using the HPLC method employed here (data not shown). The monodesamido peptide elutes after the native peptide consistent with the observations in FIG. 7. Table 5: Predicted Tryptic Peptides for VRS-317

Deamidation of rhGH is known to occur at elevated temperature with increasing pH and truncation of the XTEN domain of VRS-317, as shown in the above example, increases with decreasing pH (from pH 6 to pH 5). A formulation of pH 5.5 was selected to minimize both deamidation and truncation of the XTEN domain. At 40°C storage for periods of up to 1 month or 25°C storage for periods of up to 3 months, no deamidation of the rhGH domain of VRS-317 was detected by tryptic mapping. rhGH stored under these conditions has been observed to undergo extensive deamidation (Pearlman and Bewley, supra at 27-29).

As described earlier, another degradation pathway of rhGH is aggregation. Typically, rhGH aggregation is reduced or prevented by the addition of surfactants such as polysorbate 20 (Tween® 20) or Poloxamer 188 (Pluronic® F68). To evaluate the need for surfactants in the VRS-317 formulation, 122 mg/ml of VRS-317 was formulated in 20 mM L-histidine, 154 mM sodium chloride, pH 5.5 with and without Tween® 20 or Pluronic® F68 and stored for 4 hours while agitated at room temperature. Non-agitated and surfactant free samples served as controls. Samples were analyzed by visual inspection, absorbance at 600 nm (to monitor for turbidity), and SE-HPLC (to monitor for soluble aggregation). VRS-317 samples at 122 mg/mL remained clear and particle free after agitation with and without surfactant. No turbidity was observed (A600 < 0.01) for 122 mg/mL VRS-317 following agitation in the presence and absence of surfactant. As shown in FIG. 8, a slight increase in pre-peak species was observed for agitated VRS-317 in the absence of surfactant, slightly lower pre-peaks were seen following agitation for the product in the presence of 0.1% Pluronic® F68 and 0.1% Tween® 20, and a significant pre-peak species was seen in the presence of 0.01% Tween® 20 after agitation. Taken together, these results indicate that VRS-317 formulated as described did not significantly aggregate even in the absence of a surfactant and following agitation stress for 4 hours at room temperature.

To further evaluate the need for surfactants in the VRS-317 formulation, VRS-317 was formulated in 20 mM L-histidine, 154 mM sodium chloride, pH 5.5 with and without Tween® 20 and stored at 40°C for up to 4 weeks. As shown in Table 6, the addition of Tween® 20 was not necessary to prevent aggregation since VRS-317 did not aggregate under these conditions, likely due to the hydrophilic XTEN domains. In contrast, rhGH aggregates under similar conditions as previously reported (Cleland et al 1993, supra).

Table 6: SEC HPLC analysis of VRS-317 formulated with and without Tween® 20 and stored at 40°C for up to 4 weeks.

0.01% Tween® 20, 40°C, week 1 1.29 98.71

0.1% Tween® 20, 40°C, week 1 0.68 99.32

No Tween® 20, 40°C, week 2 0.56 98.75

0.01% Tween® 20, 40°C, week 2 0.65 99.35

0.1% Tween® 20, 40°C, week 2 0.68 98.68

No Tween® 20, 40°C, week 4 0.63 98.39

0.01% Tween® 20, 40°C, week 4 0.63 98.1

0.1% Tween® 20, 40°C, week 4 0.61 97.92

As noted above, oxidation of VRS-317 may be induced with the addition of peroxide and changes in the tryptic digest profile of methionine containing tryptic peptides can be detected after oxidation. Polysorbate and other polyether containing surfactants used in rhGH formulations to prevent aggregation are a source of oxygen free radicals that cause oxidation or rhGH (Pearlman and Bewley, supra at 26-42; Cleland et al., supra at 307-77). The optimal VRS-317 formulations do not require polysorbate (e.g. Tween® 20) or other surfactants.

VRS-317 formulated in 20 mM L-histidine, 154 mM sodium chloride, pH 5.5 stored for 24 months at 5°C, 3 months at 25°C or 1 month at 40°C does not contain detectable amounts of oxidized rhGH as measured by the tryptic mapping method noted above. In contrast, rhGH stored under these conditions with the typical surfactant required for prevention of aggregation (e.g. Tween® 20) undergoes extensive oxidation (Pearlman and Bewley, supra at 26-42; Cleland et al., supra at 307-77).

Long-Term Stability of VRS-317 Formulation Dispensed into Syringes, Cartridges, and Vials

The aqueous formulation was prepared according to the present invention containing 125 mg/ml of VRS-317 in 20 mM L-histidine and 154 mM NaCl at pH 5.5, and filled into syringes, cartridges, and vials to determine the stability of VRS-317 in solution over time.

The formulation containing VRS-317 according to present invention was dispensed into 0.1 ml and 1.0 ml syringes and stored for several months at 5°C or 25°C to determine the chemical stability of VRS-317 in the formulation over time. At the intervals shown in Table 7, samples were analyzed using SE-HPLC as described. The initial chromatogram of the formulation at month 0 showed three distinct peaks, two small pre-peaks, and a much larger main peak corresponding to VRS-317. Table 7 shows integrated percent peak areas corresponding to these three peaks, which are also plotted in FIG. 9, sampled from syringes at various times over 24 months at 5°C or 25°C. The limited degree to which the percent peak areas changed over time indicate that VRS-317 is stable and does not aggregate as an aqueous liquid formulation when stored at 5°C over a period of at least 24 months or at 25°C over a period of at least 6 months. Similar stability has been achieved up to 30 months and is expected to be achieved up to 42 months.

Table 7: SE-HPLC analysis of VRS-317 formulated according to the present invention and stored for several months at 5°C or 25°C

The formulation containing VRS-317 according to present invention was dispensed into 1.0 ml glass cartridges and stored for one month at 25°C to determine the chemical stability of VRS-317 in the formulation over time. Samples were analyzed using RP-HPLC, as described, at 2 weeks and one month as shown in Table 8. Table 8 and FIG. 10 show integrated percent peak areas corresponding to the RP-HPLC chromatogram of VRS-317 aqueous formulations sampled at 2 weeks and one month stored at 25°C. These data indicate that VRS-317 is stable as an aqueous liquid formulation for a period of weeks, even when stored only at 25°C. Table 8: RP-HPLC analysis of VRS-317 formulated according to the present invention and stored for several months at 25°C

The formulation containing VRS-317 according to present invention was dispensed into 1.0 ml vials and stored for several months at 5°C or 25°C to determine the stability of the pH of the formulation over time. The pH of these samples was tested at the intervals shown in Table 9, data also plotted in FIG. 11. As these measurements show, the pH of the formulation remained stable during storage over a period of at least 24 months, when the formulation was stored at either 5°C, or at least 6 months when the formulation was stored at 25°C. Similar stability has been achieved up to 30 months and is expected to be achieved up to 42 months.

Table 9: pH of the VRS-317 formulation according to the present invention after storage for several months at 5°C or 25°C

Claims

WHAT IS CLAIMED IS:

1. A storage stable aqueous liquid formulation comprising:

(i) about 25 mg/ml to about 150 mg/ml of a fusion protein comprising an extended recombinant polypeptide (XTEN) fused to a growth hormone (GH) sequence, wherein the GH sequence is at least 90% identical to the amino acid sequence of SEQ ID NO: 43, and

(ii) a buffer;

at a pH of about 4 to about 6, in the absence of a surfactant.

2. The aqueous liquid formulation of claim 1, wherein the formulation is free of a preservative.

3. The aqueous liquid formulation of claim 1, wherein the pH is about 5 to about

6.

4. The aqueous liquid formulation of claim 3, wherein the pH is 5.2 to 5.8.

5. The aqueous liquid formulation of claim 4, wherein the pH is about 5.5.

6. The aqueous liquid formulation of claim 1, wherein said buffer is selected from the group consisting of histidine, citrate, and succinate buffers.

7. The aqueous liquid formulation of claim 6, wherein the buffer is a histidine buffer at a concentration of about 20 mM.

8. The aqueous liquid formulation of claim 1, wherein the concentration of the fusion protein is selected from the group consisting of about 25 mg/ml, about 50 mg/ml, about 75 mg/ml, about 100 mg/ml, about 125 mg/ml, and about 150 mg/ml.

9. The aqueous liquid formulation of claim 8, wherein the concentration of the fusion protein is about 50 mg/ml.

10. The aqueous liquid formulation of claim 8, wherein the concentration of the fusion protein is about 100 mg/ml.

11. The aqueous liquid formulation of claim 8, wherein the concentration of the fusion protein is about 150 mg/ml.

12. The aqueous liquid formulation of claim 1, wherein the aqueous liquid formulation is storage stable for at least about one month at about 23±3°C.

13. The aqueous liquid formulation of claim 1, wherein the aqueous liquid formulation is storage stable for at least about 6 months to about 3 years at about 5±3°C.

14. The aqueous liquid formulation of claim 13, wherein the aqueous liquid formulation is storage stable for at least about 6 months to about 18 months at about 5±3°C.

15. The aqueous liquid formulation of claim 1, wherein the oxidation of the fusion protein is reduced as compared to the oxidation of the corresponding fusion protein formulated with a surfactant.

16. The aqueous liquid formulation of claim 1, wherein the deamidation of the fusion protein is reduced as compared to the deamidation of the corresponding fusion protein formulated at a higher pH.

17. The aqueous liquid formulation of claim 1, wherein the fusion protein does not show detectable oxidation or deamidation following incubation for about 8 hours at about 40°C.

18. The aqueous liquid formulation of claim 1, further comprising a tonicif ing agent.

19. The aqueous liquid formulation of claim 18, wherein the tonicifying agent is

NaCl.

20. The aqueous liquid formulation of claim 1 having an osmolality between about 300 mOsm to about 600 mOsm.

21. The aqueous liquid formulation of claim 1, wherein the formulation is disposed in a container containing at least one dose.

22. The aqueous liquid formulation of claim 21, wherein the container comprises a syringe.

23. The aqueous liquid formulation of claim 21, wherein the container comprises a glass cartridge.

24. The aqueous liquid formulation of claim 21, wherein the container comprises a vial.

25. The aqueous liquid formulation of claim 1 for injection into a subject with a needle.

26. The aqueous liquid formulation of claim 25, wherein the needle size is selected from the group consisting of 26 gauge, 27 gauge, 29 gauge, and 30 gauge.

27. The aqueous liquid formulation of claim 1, comprising the fusion protein according to the formula I:

(XTEN_l)_x-GH-(XTEN_2)y

wherein independently for each occurrence:

(a) x is either 0 or 1; and

(b) y is either 0 or 1, wherein x+y>l; wherein XTEN l and XTEN 2 comprise identical or different sequences.

28. The aqueous liquid formulation of claim 27, wherein the XTEN l comprises a sequence having at least 90% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOS: 18-34, 38, 39, 41, and 42.

29. The aqueous liquid formulation of claim 27, wherein the XTEN 2 comprises a sequence having at least 90% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOS: 13-17, 35, 36, 37, and 40.

30. The aqueous liquid formulation of claim 28, wherein the XTEN l comprises SEQ ID NO:39.

31. The aqueous liquid formulation of claim 29, wherein the XTEN 2 comprises

SEQ ID NO: 35.

32. The aqueous liquid formulation of claim 27, wherein XTEN l comprises SEQ ID NO: 39 and XTEN 2 comprises SEQ ID NO: 35.

33. The aqueous liquid formulation of claim l,wherein the fusion protein is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 1.

34. The aqueous liquid formulation of claim 33, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 1.

35. A method of making a storage stable aqueous liquid formulation of a fusion protein comprising an extended recombinant polypeptide (XTEN) linked to a growth hormone (GH) sequence, wherein the GH sequence is at least 90% identical to the amino acid sequence of SEQ ID NO:43, comprising mixing said fusion protein and an aqueous, pharmaceutically acceptable vehicle which includes:

i) said fusion protein at a concentration of about 25 mg/ml to about 150 mg/ml; and

ii) a buffer providing a pH of about 4 to about 6;

wherein said aqueous liquid formulation is free of a surfactant.

36. The method of claim 35 without use of a preservative.

37. A method of treating a growth-hormone related condition in a subject, comprising administering to the subject a therapeutically effective amount of the aqueous liquid formulation of claim 1, wherein the growth-hormone related condition is selected from the group consisting of growth-hormone deficiency, Turner's Syndrome, Prader-Willi Syndrome, small for gestational age (SGA), idiopathic short stature, AIDS wasting, multiple sclerosis, Crohn's disease, ulcerative colitis, and muscular dystrophy.

38. The method of claim 37, wherein the therapeutically effective amount is administered at least 48 h, or at least 72 h, or at least about 96 h, or at least about 120 h, or at least about 7 days, or at least about 14 days, at least about 21 days, or at least about 30 days between consecutive doses.

39. The method of claim 37, wherein growth-hormone related condition is growth-hormone deficiency.

40. The method of claim 37, wherein growth-hormone related condition is Turner's Syndrome.

41. The method of claim 37, wherein growth-hormone related condition is Prader- Willi Syndrome.

42. The method of claim 37, wherein growth-hormone related condition is idiopathic short stature.

43. The method of claim 37, wherein growth-hormone related condition is small for gestational age (SGA).

44. The method of claim 37, wherein the growth-hormone related condition is selected from the group consisting of AIDS wasting, multiple sclerosis, Crohn' s disease, ulcerative colitis, and muscular dystrophy.

45. The use of a therapeutically effective amount of the aqueous liquid

formulation of claim 1 in the preparation of a medicament for treating a growth-hormone related condition, wherein the growth-hormone related condition is selected from the group consisting of growth-hormone deficiency, Turner's Syndrome, Prader-Willi Syndrome, small for gestational age (SGA), idiopathic short stature, AIDS wasting, multiple sclerosis, Crohn's disease, ulcerative colitis, and muscular dystrophy.

46. The aqueous liquid formulation of claim 1 for use in treating a growth- hormone related condition, wherein the growth-hormone related condition is selected from the group consisting of growth-hormone deficiency, Turner's Syndrome, Prader-Willi Syndrome, small for gestational age (SGA), idiopathic short stature, AIDS wasting, multiple sclerosis, Crohn's disease, ulcerative colitis, and muscular dystrophy.