WO2014096440A2

WO2014096440A2 - Composition

Info

Publication number: WO2014096440A2
Application number: PCT/EP2013/077869
Authority: WO
Inventors: Jens Thostrup BUKRINSKI; Mette-Marie List JENSEN; Sachin DUBEY
Original assignee: Novozymes Biopharma Dk A/S
Priority date: 2012-12-21
Filing date: 2013-12-20
Publication date: 2014-06-26
Also published as: WO2014096440A3

Abstract

The present invention concerns compositions comprising highly purified albumin suitable for preventing and/or reducing of peptide fibrillation.

Description

Composition

Reference to a Sequence Listing

This application contains a Sequence Listing in computer readable form, which is incorporated herein by reference.

Field of invention

The present invention relates to a composition comprising Human Serum Albumin (HSA) and its use for preventing or reducing self-association and aggregation of peptide based drugs, and in particular the formation of amyloids, visible and sub-visible particles in insulin.

Background of invention

Aggregation of peptides in peptide drug and vaccine formulation is a source of dosage form instability and also undesirable immunogenic responses. Maximizing the stability of a therapeutic peptide is important for providing a safe drug product with an appropriate shelf-life. Maximizing the stability results in a more convenient dosage form and, eventually, in better patient compliance. Maintaining the physical stability, e.g. a non-aggregated form of, the peptide with the correct three-dimensional structure is essential for interactions with the therapeutic target, as well as for ensuring efficacy and avoiding immunological reactions. In addition, protein instability during manufacture, final drug formulation, and storage can influence the product yield and shelf-life of the peptide.

To achieve a stable pharmaceutical drug product, excipients are sometimes added to the active peptide drug substance. Human Serum Albumin (HSA) as well as a variety of sugars, salts, amino acids and detergents has been found to be useful excipients in both peptide drug and vaccine formulations.

The biologically active form of insulin is monomeric, but the monomer is labile and prone to aggregation. Insulin, however, exists in equilibrium between a monomeric, dimeric and hexameric form and some modern insulin analogues even exist as multimers of hexamers. The equilibrium between the quaternary states of insulin is shifted towards the hexameric state upon addition of Zn²⁺. The hexamer is stabilized by Zn²⁺ ions, thus forming a hexamer of insulin (PDB ID: 1AI0; Chang et al (1997) Biochemistry 36 (31 ): 9409-22). The hexamer is less prone to aggregation and fragmentation but it is also biologically inactive. Accordingly, the hexamer has to dissociate into monomers to be active and is thus a sustained release form.

The diabetic patient needs two insulin presentations - A) a basal insulin (such as levemir (Novo Nordisk) or Lantus (Sanofi Aventis)) that will keep a stable blood sugar level for a prolonged period such as a day; and B) a fast acting insulin (such as Novo Rapid (Novo Nordisk) and Humalog (Ely Lily)) that can be administered in conjunction with each meal to facilitate the regulation of the increased glucose load associated with the food intake. In order to prepare a fast acting insulin formulation which comprises zinc ions for stabilization of the hexamer, it is necessary to modify the amino acid sequence in order to facilitate a more rapid dissociation of the hexamer.

Modern insulin analogues mentioned above are very expensive and therefore are often out of reach for many consumers in developing countries. Non modified human insulin however is inexpensive. Thus it is desirable to provide an alternative to zinc ions for stabilizing human insulin, e.g. in monomer form, in order to meet an unmet medical need and thus to create an opportunity to increase the value of life for millions of diabetics in third world countries.

Serum derived HSA has previously been used to stabilize peptides both in solution and in freeze-dried state. As the most abundant protein in human plasma, the potential for HSA to illicit an immunogenic response is minimal, making it an ideal excipient candidate. However, serum derived HSA has the disadvantage of being derived from donated human blood with the attendant risk of contamination with infection agents. Hence recombinant Human Serum Albumin (rAlbumin) has been suggested as substitute for serum derived HSA in the stabilization and formulation of peptides.

rAlbumin can be produced in recombinant microorganisms such as the yeasts Saccharomyces cerevisiae (Sleep et al. (1991 ) BioTechnology 9:183-187), Kluyveromyces lactis (Fleer et at. (1991 ) BioTechnology 9:968-975) or Pichia pastoris (EP510693) or bacteria such as E. coli (Latta et al (1987) Ann. Hematol 68:S21-S24).

Alternatively, the rAlbumin can be derived from transgenic plants or animals (Sijmons et al (1990) BioTechnology 8:217-221 ; Shani et al (1992) Transgenic Research 1 :195-208).

rAlbumin has been formulated with recombinant Factor VIII (Res. Disci. (1995) 376(08): 516). WO 2003/066681 suggests retained activity of Factor VIII in combination with rAlbumin over time when compared to control.

Rasmussen et al (2010) Pharmaceutical Research 27:1337-1347 demonstrates results from a study using alpha-crystallin for stabilizing insulin. The composition also comprises HSA as a control stabilizer, however the stabilizing effect of HSA on insulin is not particularly good as compared to alpha-crystallin. The HSA sample is used without further purification.

W01992/019260 discloses that albumin increases the stability of insulin up to a certain concentration but that the stabilizing effect starts to decrease at 10mg/ml albumin.

Summary of invention

The present inventors have prepared a composition comprising recombinant human albumin (HSA) to prevent or reduce self-association of peptide/peptide-based drugs and especially the to prevent or reduce formation of fibril aggregates in insulin, glucagon, GLP-1 and analogs thereof, GLP-2 and analogs thereof and a HIV fusion inhibitor.

In a main aspect, the invention concerns a composition comprising from 0.01 mg/ml to 300 mg/ml peptide (such as from 10 to 100 amino acids) and from 0.01 mg/ml to 500 mg/ml albumin, and wherein the composition comprises less than or equal to 25 mM small hydrophobic molecules and/or 25 mM fatty acids such as octanoate and/or less than 0.001 % (w/v) detergent such as polysorbate 80 and/or less than 5 mM free amino acids. A composition comprising less than or equal to 25 mM octanoate and/or less than 0.001 % (w/v) polysorbate 80 is preferred.

In another main aspect the invention concerns use of the above composition for preventing and/or reducing formation of peptide fibrils.

In yet another aspect the invention concerns use of a highly purified albumin composition for preventing and/or reducing formation of insulin fibrils in an essentially zinc ion free formulation of insulin.

The invention in another main aspect also concerns a method of stabilizing an essentially zinc ion free insulin composition comprising dissolving zinc ion free insulin in an aqueous solution of highly purified albumin, wherein the aqueous solution is essentially detergent free.

The invention in another main aspect concerns a composition which is essentially free of zinc comprising:

a. from 0.01 mg/ml to 300 mg/ml insulin or insulin analog;

b. from 0.01 mg/ml to 500 mg/ml albumin

wherein the composition comprises less than or equal to 25 mM octanoate and/or less than 0.001 % (w/v) polysorbate 80.

The invention in another main aspect concerns a method of preventing and/or reducing formation of peptide fibrils in an aqueous solution, the method comprising dissolving the peptide in an aqueous solution of albumin, wherein the aqueous solution is essentially free of amphiphilic compounds.

The invention furthermore concerns a method of preventing and/or reducing formation of peptide fibrils, the method comprising preparing a composition as defined herein above, wherein the peptide referred to in said composition is the peptide to be stabilized in order to avoid formation of peptide based fibrils.

Description of Drawings

Figure 1 : Time dependent thioflavin T (ThT) binding to fibrils formed by incubation of human insulin at 35 °C with orbital shaking. The assay was performed at pH 7.0 with 1 mM EDTA in all samples (to immobilize the Zn²⁺ present and thereby favor the monomeric state of insulin), 1 mM ThT and 0.5 mg/ml (0.086 mM) insulin was prepared in a sample volume of 200 μΙ_ and all results are a mean of three measurements. The concentration of albumin when added was 5.74 mg/ml (0.086 mM) resulting in a molar ratio of 1 :1 insulin:rAlbumin.

Figure 2: Time dependent thioflavin T (ThT) binding to fibrils formed by incubation of human insulin at 35 °C with orbital shaking. The assay was performed at pH 8.0 with 1 mM EDTA in all samples (to immobilize the Zn²⁺ present and thereby favor the monomeric state of insulin), 1 mM ThT and 0.5 mg/ml (0.086 mM) insulin was prepared in a sample volume of 200 μΙ_ and all results are a mean of three measurements. The concentration albumin when added was 5.74 mg/ml (0.086 mM) resulting in a molar ratio of 1 :1 insulin:rAlbumin.

Figure 3 - Affect of albumin (Albix) concentration on the stability of A & B: human insulin

(HI) and, C&D: insulin analog (lispro), as studied by ThT assay. HI: human insulin. Albix: a formulation of albumin comprising maximum 2 mM octanoate and substantially free of detergent.

Figure 4 - Affect of albumin (Albix) concentration on the stability of human glucagon (G) as studied by SE-HPLC. "Phos": phosphate buffer. HI: human insulin. Albix: a formulation of albumin comprising maximum 2 mM octanoate and substantially free of detergent.

Figure 5 - Affect of polysorbate 80 (Tween 80), polysorbate 20 (Tween 20), octanoate and albumin (Albix) on the stability of glucagon ("G") as studied by SE-HPLC. "Phos": phosphate buffer.

Figure 6 - Affect of octanoate (Oct) and polysorbate 80 (T80) on the ability of albumin to stabilize glucagon as studied by SE-HPLC. G: glucagon, Albucult: a formulation of albumin comprising nominally 8 mM octanoate and maximum 50 mg/L polysorbate 80; Albix: a formulation of albumin comprising maximum 2 mM octanoate and substantially free of detergent, Phos: phosphate buffer. Oct: octanoate. T80: polysorbate 80.

Figure 7: Affect of buffer on the ability of albumin (Albix) to stabilize glucagon (G) as studied by SE-HPLC. G: glucagon; Phos: phosphate buffer; Cit: citrate buffer; His: histidine buffer.

Figure 8: Affect of octanoate, polysorbate 20 (T20) and polysorbate 80 (T80) on the ability of albumin to stabilize glucagon-like peptide 2 analog (teduglutide) as studied by ThT assay. GLP2 / GLP: GLP-2 analog; Albucult: a formulation of albumin comprising nominally 8 mM octanoate and maximum 50 mg/L polysorbate 80; Albix: a formulation of albumin comprising maximum 2 mM octanoate and substantially free of detergent. T20: polysorbate 20. T80: polysorbate 80.

Figure 9: Affect of albumin (Albix) concentration on the stability of glucagon-like peptide 2 analog (teduglutide) as studied by ThT assay. GLP2 / GLP: GLP-2 analog; Albix: a formulation of albumin comprising maximum 2 mM octanoate and substantially free of detergent.

Figure 10: Affect of amino acids on the ability of albumin to stabilize glucagon-like peptide 2 analog (teduglutide) as studied by ThT assay. GLP2 / GLP: GLP-2 analog; Albix: a formulation of albumin comprising maximum 2 mM octanoate and substantially free of detergent.

Figure 1 1 : Affect of albumin concentration on the stability of an HIV fusion inhibitor (enfuvirtide) as studied by ThT assay. Albix: a formulation of albumin comprising maximum 2 mM octanoate and substantially free of detergent. Detailed description of the invention

Definitions

Albumin: HSA is the general term referring to serum derived human serum albumin. rAlbumin is a general term referring to presentations of recombinant human serum albumin including both stripped rAlbumin prepared as described below and the known commercial presentations Recombumin^® Alpha (formerly Albucult^®), Recombumin® Prime (formerly Recombumin^®) and Albix™ (all from Novozymes Biopharma DK A S) used for comparative studies as is, i.e. without further preparation. The albumin according to the present invention may comprise native sequence HSA such as SEQ ID NO: 1 or a variant or fragment thereof.

The term "albumin" means a protein having the same and/or very similar three dimensional (tertiary) structure as HSA or HSA domains and has similar properties to HSA or to the relevant domains. Similar three dimensional structures are for example the structures of the albumins from the species mentioned herein. Some of the major properties of albumin are i) its ability to regulate plasma volume (oncotic activity), ii) a long plasma half-life of around 19 days ± 5 days, iii) binding to FcRn, iv) ligand-binding, e.g. binding of endogenous molecules such as acidic, lipophilic compounds including bilirubin, fatty acids, hemin and thyroxine (see also Table 1 of Kragh-Hansen et al, 2002, Biol. Pharm. Bull. 25, 695, hereby incorporated by reference), v) binding of small organic compounds with acidic or electronegative features e.g. drugs such as warfarin, diazepam, ibuprofen and paclitaxel (see also Table 1 of Kragh-Hansen et al, 2002, Biol. Pharm. Bull. 25, 695, hereby incorporated by reference). Not all of these properties need to be fulfilled in order to characterize a protein or fragment as an albumin. If a fragment, for example, does not comprise a domain responsible for binding of certain ligands or organic compounds the variant of such a fragment will not be expected to have these properties either.

Amphiphilic compounds: The term 'amphiphilic' compound or molecule as used herein refers to a chemical compound possessing both hydrophilic and lipophilic properties. As used herein, amphiphilic is to be understood as including detergents, fatty acids as well as phospholipids.

Fragment: The term 'fragment' means from 20, 30, 40, 50, 60, 70, 80, or 90 to 30, 40, 50, 60, 70, 80, 90, 95% of the length of the peptide from which the fragment is derived. It is preferred that the fragment has at least 50%, more preferably at least 60, 70, 80, 90, 95, 99 or 100% of the functional activity of the peptide from which it is derived.

Free amino acid: The term ree amino acid' means an amino acid which is not bound, e.g. covalently, linked to any other amino acids, e.g. not part of dipeptide, a peptide or a protein.

Peptide: In the following, peptides are meant to be small proteins constituted by one peptide chain or by two or more covalently linked peptide chains. A peptide may have a total number of amino acids above 5 amino acids and less than 200, especially from 10 to 100 amino acids. Peptides are likewise meant to include homo multimers comprising 2 or more peptide sub-units with same amino acid sequence (such as an insulin hexamer) as well as hetero multimers comprising 2 or more peptide sub-units of different amino acid sequences (such as an insulin hexamer comprising insulin of two different sequences e.g. a fast acting variant and a long acting variant).

Prevention (or inhibition): In relation to aggregates e.g. fibrils, the term 'prevention' or inhibition' means hindering the formation of aggregates, e.g. fibrils. Prevention or inhibition may be complete, e.g. no aggregates are formed. Prevention or inhibition may be partial, e.g. fewer aggregates are formed compared to a reference composition or the aggregates formed may be incomplete e.g. an aggregate intermediate between a monomer (non-aggregate) and a quaternary structure (e.g. a fibril or hexamer) may be formed. Prevention or inhibition may be slower for a test composition than for a reference composition, e.g. the rate of aggregate, such as fibril, formation may be decreased. A 'reference' composition may be a composition in which albumin or stripped albumin is absent. A 'reference' composition may be a composition which includes a lower or higher (particularly higher) level of one or more (several) components such as fatty acid (particularly octanoate), detergent (particularly polysorbate 80) or salt (particularly NaCI) compared to the test composition. For example, aggregation may be at most 90, 80, 70, 60, 50, 40, 30, 20, 10, 5, 4, 3, 2, 1 , 0.1 , 0.01 % of the aggregates formed in a reference composition. A reference composition may comprise 0.05 % (w/v) polysorbate 80, polysorbate 20 and/or total detergent. A reference composition may comprise 0.8 mM octanoate and/or total fatty acid. By 'total fatty acid' is meant all types of fatty acids.

Prevention (or inhibition) or reduction of aggregation may be measured following exposure of the composition to a stress test such as exposure to shaking and/or heating (e.g. Example 2) and/ prolonged exposure to hydrophobic surfaces or by repeated freezing and thawing. Quantitation of aggregation by done by a suitable assay such as the ThT assay described in Example 1 , Example 2 or Example 3a. The quantification of aggregates can alternatively be performed using size exclusion chromatography e.g. as described in the size exclusion high performance liquid chromatography (SE-HPLC) assay of Example 3b, of different types of light scattering techniques based dynamic or static light scattering or by different microscopic techniques such as light microscopy and micro flow imaging.

Reduction: In relation to aggregates e.g. fibrils, the term 'reduction' means removal of existing aggregates, e.g. fibrils.

Sequence Identity: The relatedness between two amino acid sequences is described by the parameter "sequence identity". For purposes of the invention, the degree of sequence identity between two amino acid sequences is determined using the Needleman-Wunsch algorithm (Needleman and Wunsch, 1970, J. Mol. Biol. 48: 443-453) as implemented in the Needle program of the EMBOSS package (EMBOSS: The European Molecular Biology Open Software Suite, Rice er a/., 2000, Trends Genet. 16: 276-277), preferably version 3.0.0 or later, more preferably version 5.0.0 or later. The optional parameters used are gap open penalty of 10, gap extension penalty of 0.5, and the EBLOSUM62 (EMBOSS version of BLOSUM62) substitution matrix. The output of Needle labelled "longest identity" (obtained using the -nobrief option) is used as the percent identity and is calculated as follows: (Identical Residues x 100)/(Length of Alignment - Total Number of Gaps in Alignment)

Composition suitable for stabilizing peptides in aqueous solution

Aggregation of peptides in peptide drug formulation is a source of dosage form instability and may also result in undesirable immunogenic responses. It is therefore desirable to maximize stability of a therapeutic peptide in order to provide a safe drug product with an appropriate shelf-life. For this purpose the present inventors have formulated a composition which has been shown to stabilize peptides in aqueous solution.

In a main aspect the present invention concerns a composition comprising from 0.01 mg/ml to 300 mg/ml peptide (such as from 10 to 100 amino acids) and from 0.01 mg/ml to 500 mg/ml albumin, and wherein the composition comprises less than or equal to 25 mM small hydrophobic molecules and/or 25 mM fatty acids and/or less than 0.001 % (w/v) detergent and/or less than 5 mM free amino acids. A composition comprising less than or equal to 25 mM octanoate and/or less than 0.001 % (w/v) polysorbate 80 is preferred.

For example, the composition may comprise from 0.01 mg/ml to 300 mg/ml peptide

(such as a peptide having a size of from 10 to 100 amino acids) and from 0.01 mg/ml to 500 mg/ml albumin; wherein the composition comprises less than or equal to 25 mM octanoate and/or less than 0.001 % (w/v) polysorbate 80.

For example, the composition may comprise from 0.01 mg/ml to 300 mg/ml peptide (such as a peptide having a size of from 10 to 100 amino acids) and from 0.01 mg/ml to 500 mg/ml albumin; wherein the composition comprises less than or equal to 25 mM fatty acid and/or less than 0.001 % (w/v) detergent.

In one embodiment, the peptide is a non-lipopeptide.

In one embodiment the composition according to the present invention comprises a molar ratio of peptide to albumin ranging from 1 part peptide to 2000 parts albumin (1:2000) to 3000 parts peptide to 1 part albumin (1:3000) such as from about 1:1000, 1:500, 1:250, 1:100, 1:80, 1:50, 1:36, 1:40, 1:25, 1:20, 1:10, 1:7, 1:5.2, 1:5, 1:4.3, 1:4, 1:3.7, 1:3.3, 1.3, 1:2, 1:1.15, 1:1, 1.15:1, 1.4:1, 1.5:1, 2:1, 3:1, 4:1, 5:1, 10: 1, 50:1, 100:1, 200:1, 250:1, 500:1, 750:1, 1000:1, or 1500:1 to about 1:500, 1:250, 1:100, 1:80, 1:50, 1:40, 1:36, 1:25, 1:20, 1:10, 1:7, 1:5.2, 1:5, 1:4.3, 1:4, 1:3.7, 1:3.3, 1.3, 1:2, 1:1.15, 1:1, 1.15:1, 1.4:1, 1.5:1, 2:1, 3:1, 4:1, 5:1, or 10:1,50:1, 100:1,200:1,250:1,500:1,750:1, 1000:1, 1500:1 or 2500:1.

In another embodiment the composition according to the present invention comprises a molar ratio ranging from 0.01 to 500 mg/ml albumin and from 1 to 100 mg/ml peptide.

In another embodiment the composition according to the present invention comprises a molar ratio ranging from 1 to 250 mg/ml albumin and from 1 to 100 mg/ml peptide.

In another embodiment the composition according to the present invention comprises a molar ratio ranging from 1 to 100 mg/ml albumin and from 1 to 100 mg/ml peptide.

In another embodiment the composition according to the present invention comprises a molar ratio ranging from 1 to 30 mg/ml albumin and from 0.01 to 100 mg/ml peptide. Albumin may be present in the composition of the present invention in a concentration of from 0.01 mg/ml to 300 mg/ml, such as from about 0.01 , 0.76 1 , 1 .3, 1.9, 2, 3.7, 5, 9.5, 10, 1 1.1 , 15, 18.5, 20, 30, 40, 50, 100, 200 to about 0.76, 1 , 1.3, 1 .9, 2, 3.7, 5, 9.5, 10, 11.1 , 15, 18.5, 20, 30, 40, 50, 100, 200, 300 mg/ml.

Albumin as referred to herein is preferably HSA (SEQ ID NO: 2). In certain embodiments the albumin may be selected from the group consisting of SEQ ID NO. 10: Albumin - Pan troglodytes; SEQ ID NO. 1 1 : Albumin - Macaca mulatta; SEQ ID NO. 12: Albumin - Mesocricetus auratus; SEQ ID NO. 13: Albumin - Cavia porcellus; SEQ ID NO. 14: Albumin - Mus musculus; SEQ ID NO. 15: Albumin - Rattus norvegicus; SEQ ID NO. 16: Albumin - Bos taurus; SEQ ID NO. 17: Albumin - Equus caballus; SEQ ID NO. 18: Albumin - Equus asinus; SEQ ID NO. 19: Albumin - Oryctolagus cuniculus; SEQ ID NO. 20: Albumin - Capra hircus; SEQ ID NO. 21 : Albumin - Ovis aries; SEQ ID NO. 22: Albumin - Canis lupus familiaris; SEQ ID NO. 23: Albumin - Gallus gallus and SEQ ID NO. 24: Albumin - Sus scrofa, or a fragment or variant of any one of said SEQ ID NOs 2, 10, 1 1 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23 and 24.

The variants of the herein mentioned albumins or peptides may be any variant with retained biological activity. Thus the variant can be an amino acid sequence being at least 50%, preferably at least 60%, preferably at least 70%, preferably at least 75%, preferably at least 80%, preferably at least 85%, preferably at least 90%, preferably at least 95%, preferably at least 98%, more preferably at least 99% and most preferably at least 99.5% identical to any one of the above mentioned albumins or peptides. The albumin variant or peptide variant may differ from HSA or one of the above mentioned peptides by one or more (several) amino acids such as from 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, or 20 amino acids to 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20 or 25 amino acids. Peptides

It is a main object of the present invention to prevent or reduce peptide aggregation and/or formation of peptide fibrils. Hence it is preferred that the peptide of the above defined composition is essentially in non-fibril form.

The peptide may or may not be a lipopeptide. Non-lipopeptides are preferred. That is peptides containing only natural amino acids and non-natural amino acids are preferred.

In one embodiment the composition of the present invention is formulated for purposes of stabilizing one peptide type, however in certain embodiments the composition may comprises two or more different peptides and provide appropriate stabilization for both (all) types of peptides included in the formulation. A peptide may comprise two or more different peptide chains, e.g. comprises an A chain and a B chain. Examples of peptides comprising A and B chains include insulin and insulin analogs.

The peptide to be stabilized by formulation in the composition according to the present invention can be any peptide which aggregates or forms fibrils in aqueous solution. In one embodiment the peptide is selected from the group consisting of insulin (as described herein), glucagon-like peptides including GLP1 (SEQ ID NO: 5), GLP2 (SEQ ID NO: 6), human growth hormone (SEQ ID NO: 7), glucagon (SEQ ID NO: 8), GLP-1 analogs (e.g. SEQ ID NO: 27), GLP-2 analogs (e.g. SEQ ID NO: 28), HIV fusion inhibitors (e.g. SEQ ID NO: 29), cytokines such as interleukins, interferons, chemokines and other peptide hormones as well as fragments or variants of any one of said peptides.

The term 'insulin' as used herein may comprise a native insulin, or an analog or fragment therefore. Insulin may comprise one chain (e.g. a single chain insulin) or two chains such as A and B. Native human insulin is generated from immature SEQ ID NO: 2, the C-chain is excised to generate an A chain and a B chain which become linked together by two disulfide bonds. It is preferred that the single chain or the A chain has at least 60, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99, 99.5 % identity or 100% identity to SEQ ID NO: 3 or SEQ ID NO: 25. It is preferred that the A chain comprises 19, 20, 21 , 22 or 23 amino acids, preferably 21 . It is preferred that the single chain or B chain has at least 60, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99, 99.5 % identity or 100% identity to SEQ I D NO: 4 or SEQ I D NO: 26. It is preferred that the B chain comprises 28, 29, 30, 31 , 32 or 33 amino acids, preferably 30. The insulin may comprise SEQ ID NO: 3 and SEQ I D NO: 4 or variants thereof or SEQ I D NO: 25 and SEQ ID NO: 26 or variants thereof.

The interleukin to be formulated as the peptide of the present invention may be selected from the group consisting of IL-1 , IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-1 1 , IL-12, IL-13, IL-14, I L-15, IL-16, I L-17, IL-18, I L-19, IL-20, I L-21 , I L-22, IL-23, I L-24, IL-25, I L-26, IL-27, IL-28, IL-29, IL-30, IL-31 , IL-32, IL-33, IL-34, IL-35, IL-36 and fragments or variants of any one of said peptides.

The interferon to be formulated as the peptide of the present invention may be selected from the group consisting of I FNA1 , IFNA2, I FNA4, I FNA5, I FNA6, IFNA7, I FNA8, I FNA10, I FNA13, IFNA14, I FNA16, I FNA17, IFNA21 , I FNB1 , IFNW, I FNE1 , IFNK and fragments or variants of any one of said peptides.

The cytokine to be formulated as the peptide of the present invention may be selected from the group consisting of CC chemokines, CXC chemokines, C chemokines and CX3C chemokines and fragments or variants of any one of said peptides.

In one embodiment the CC chemokines are selected from the group consisting of CCL1 ,

CCL2, CCL3, CCL4, CCL5, CCL6, CCL7, CCL8, CCL9/CCL10, CCL1 1 , CCL12, CCL13, CCL14, CCL15, CCL16, CCL17, CCL18, CCL19, CCL20, CCL21 , CCL22, CCL23, CCL24, CCL25, CCL27, CCL28 and fragments or variants of any one of said peptides.

In one embodiment the CXC chemokines are selected from the group consisting of CXCL1 , CXCL2, CXCL3, CXCL4, CXCL5, CXCL6, CXCL7, CXCL8, CXCL9, CXCL10, CXCL1 1 , CXCL12, CXCL13, CXCL14, CXCL15, CXCL16, CXCL17 and fragments or variants of any one of said peptides.

In one embodiment the C chemokines are selected from the group consisting of C chemokines: XCL1 , XCL2 and fragments or variants of any one of said peptides. In one embodiment the CX3C chemokine is CX3CL1 and fragments or variants of any one of said peptide.

The peptide can also be a peptide hormone selected from the group consisting of Amylin, Antimullerian hormone, Adiponectin, Adrenocorticotropic hormone, Angiotensinogen, Angiotensin, Antidiuretic hormone, Atrial-natriuretic peptide, Brain natriuretic peptide, Calcitonin, Cholecystokinin, Corticotropin-releasing hormone, Enkephalin, Endothelin, Erythropoietin, Follicle-stimulating hormone, Galanin, Gastrin, Ghrelin, Glucagon, Gonadotropin-releasing hormone, Growth hormone-releasing hormone, Human chorionic gonadotropin, Human placental lactogen, Growth hormone, Inhibin, Insulin-like growth factor, Leptin, Lipotropin, Luteinizing hormone, Melanocyte stimulating hormone, Melanocyte stimulating hormone, Motilin, Orexin, Oxytocin, Pancreatic polypeptide, Parathyroid hormone, Prolactin, Prolactin releasing hormone, Relaxin, Renin, Secretin, Somatostatin, Thrombopoietin, Thyroid-stimulating hormone, Thyrotropin-releasing hormone and fragments or variants of any one of said peptides.

The peptide may also be selected from the group consisting of Acylation stimulating protein, Adipokine, Albinterferon, Colony-stimulating factor, Gc-MAF, Granulocyte colony- stimulating factor, Granulocyte macrophage colony-stimulating factor, Hepatocyte growth factor, Leukemia inhibitory factor, Leukocyte-promoting factor, Lymphokine, Lymphotoxin, Lymphotoxin alpha, Lymphotoxin beta, Macrophage activating factor, Macrophage inflammatory protein, Monokine, Myokine, Oncostatin M, Oprelvekin, Platelet factor 4, Promegapoietin, Stromal cell- derived factor-1 , Tumor necrosis factor-alpha and fragments or variants of any one of said peptides.

In one embodiment the peptide formulated with the composition according to the present invention is selected from the group consisting of insulin, GLP1 , GLP2, glucagon, human growth hormone, cytokines and other peptide hormones as well as fragments or variants of any one of said peptides.

The variants of the above mentioned peptides may be any variant with retained biological activity. Thus the variant can be an amino acid sequence being at least 50%, preferably at least 60%, preferably at least 70%, preferably at least 75%, preferably at least 80%, preferably at least 85%, preferably at least 90%, preferably at least 95%, preferably at least 98%, more preferably at least 99% and most preferably at least 99.5% identical to any one of the above mentioned peptides or polypeptides. The albumin variant or peptide variant may differ from HSA or one of the above mentioned peptides by one or more (several) amino acids such as from 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, or 20 amino acids to 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20 or 25 amino acids.

The peptide of the present composition may be any peptide which forms fibrils or aggregates in an aqueous and/or suspension solution including full length or fragments of the above mentioned peptides, provided that the fragments retain a substantial part of their biological activity. In one embodiment the peptide formulated in the composition according to the present invention the peptide comprises less than 200 amino acid residues, such as less than 195 amino acid residues, such as less than 190 amino acid residues, such as less than 185 amino acid residues, such as less than 180 amino acid residues, such as less than 175 amino acid residues, such as less than 160 amino acid residues, such as less than 150 amino acid residues, such as less than 140 amino acid residues, such as less than 130 amino acid residues, such as less than 125 amino acid residues, such as less than 120 amino acid residues, such as less than 1 15 amino acid residues, such as less than 1 10 amino acid residues, such as less than 105 amino acid residues, such as less than 100 amino acid residues, such as less than 95 amino acid residues, such as less than 90 amino acid residues, such as less than 85 amino acid residues, such as less than 80 amino acid residues, such as less than 75 amino acid residues, such as less than 70 amino acid residues, such as less than 65 amino acid residues, such as less than 60 amino acid residues, such as less than 55 amino acid residues, such as less than 50 amino acid residues, such as less than 45 amino acid residues, such as less than 40 amino acid residues, such as less than 35 amino acid residues, such as less than 30 amino acid residues, such as less than 25 amino acid residues, such as less than 20 amino acid residues, such as less than 15 amino acid residues. A peptide of from 10 to 100 amino acid residues is preferred, particularly a peptide residue of from 10 to 60 amino acid residues. Non-lipopeptides, as disclosed herein, are also preferred.

In one embodiment the composition according to the present invention comprises from at least 0.01 , such as from at least 1 , such as from at least 5, such as from at least 10, such as from at least such as from at least 20, such as from at least 30, such as from at least 40 to 1 , such as to 5, such as to 10, such as to 20, such as to 30, such as to 40, such as to 50, such as to 90 mg/ml or more than 100 mg/ml peptide.

In one embodiment the composition according to the present invention consists essentially of albumin and insulin.

In one embodiment the composition according to the present invention consists essentially of albumin and glucagon or analog thereof.For example, the composition according to the present invention may consist essentially of albumin and GLP-1 or analog thereof (e.g. liraglutide). For example, the composition according to the present invention may consist essentially of albumin and GLP-2 or analog thereof (e.g. teduglutide).

In one embodiment the composition according to the present invention consists essentially of albumin and HIV fusion inhibitor (e.g. enfurviride).

Detergent

Detergents can be classified into four groups, depending on the electrical charge - anionic detergents, cationic detergents, non-ionic detergents and zwitterionic detergents.

Typical anionic detergents include alkylbenzenesulfonates. The alkylbenzene portion of these anions is lipophilic and the sulfonate is hydrophilic. Examples of types of anionic detergents include branched sodium dodecylbenzenesulfonate, linear sodium dodecylbenzenesulfonate, and soap. In one embodiment, the composition of the present invention comprises less than 0.01 , preferably less than 0.001 , more preferably less than 0.0001 % (w/v) anionic detergent.

Cationic detergents are similar to the anionic detergents, with a hydrophobic component, but, instead of the anionic sulfonate group, the cationic surfactants have quaternary ammonium (i.e. positively charged group) as the polar moiety.

In one embodiment, the composition of the present invention comprises less than 0.01 , preferably less than 0.001 %, more preferably less than 0.0001 (w/v) cationic detergent.

Zwitterionic detergents possess a net zero charge arising from the presence of equal numbers of +1 and -1 charged chemical groups. One example of a zwitterionic detergent is CHAPS (3-[(3-Cholamidopropyl)dimethylammonio]-1 -propanesulfonate). In one embodiment, the composition of the present invention comprises less than 0.001 % (w/v) zwitterionic detergent and may be essentially free of zwitterionic detergents.

Non-ionic detergents are characterized by their uncharged, hydrophilic headgroups. Typical non-ionic detergents are based on polyoxyethylene or a glycoside. Common examples of the former include polysorbate 80 (e.g. Tween^®), 4-(1 ,1 ,3,3-Tetramethylbutyl)phenyl- polyethylene glycol, t-Octylphenoxypolyethoxyethanol (e.g. Triton^® X-100), and the Brij^® series. These materials are also known as ethoxylates or PEGylates. Glycosides have a sugar as their uncharged hydrophilic head-group. Examples include octyl-thioglucoside and maltosides. Hydroxyethylglucamide (HEGA) and methylglucamide (MEGA) series detergents are similar, possessing a sugar alcohol as the head-group.

In one preferred embodiment, the composition of the present invention comprises less than 0.01 , preferably less than 0.001 , more preferably less than 0.0001 % (w/v) nonionic detergent. In a further embodiment, the composition of the present invention comprises less than 0.01 , preferably less than 0.001 %, more preferably less than 0.0001 (w/v) polysorbate 80 and may be essentially free of polysorbate 80. For example, the composition may comprise less than or equal to 3.325^*10^"4 % (w/v) polysorbate 80 or 20, such as less than or equal to 2.85^*10^"4 % (w/v) polysorbate 80 or 20, such as less than or equal to 2.375^*10^"4 % (w/v) polysorbate 80 or 20, such as less than or equal to 1.425^*10^"4 % (w/v) polysorbate 80 or 20, such as less than or equal to 9^*10^"5 % (w/v) polysorbate 80 or 20, such as less than or equal to 6.625^*10^"5 % (w/v) polysorbate 80 or 20, such as less than or equal to 5.7^*10^"5 % (w/v) polysorbate 80 or 20, such as less than or equal to 5^*10^"5 % (w/v) polysorbate 80 or 20, such as less than or equal to 4.75^*10^"5 % (w/v) polysorbate 80 or 20, such as less than or equal to 4.5^*10^"5 % (w/v) polysorbate 80 or 20, such as less than or equal to 4.75^*10^"5 % (w/v) polysorbate 80 or 20, such as less than or equal to 2.85^*10^"5 % (w/v) polysorbate 80 or 20, such as less than or equal to 2.5^*10^"5 % (w/v) polysorbate 80 or 20, such as less than or equal to 1.9^*10^"5 % (w/v) polysorbate 80 or 20, such as less than or equal to 1.8^*10^"5 % (w/v) polysorbate 80 or 20, such as less than or equal to 1 ^*10^"5 % (w/v) polysorbate 80 or 20, such as less than or equal to 9.5^*10^"6 % (w/v) polysorbate 80 or 20, such as less than or equal to 9^*10^"6 % (w/v) polysorbate 80 or 20, such as less than or equal to 5^*10^"6 % (w/v) polysorbate 80 or 20, most preferably substantially free of polysorbate 80 or 20.

In another embodiment, the composition of the present invention comprises less 0.01 , preferably less than 0.001 , more preferably less than 0.0001 % (w/v) polysorbate 20 and may be free of polysorbate 20. In another embodiment, the composition of the present invention comprises less than 0.01 , preferably less than 0.001 , more preferably less than 0.0001 % (w/v) poloxamer and may be free of poloxamer.

In one embodiment the composition of the present invention comprises from 0.001 , such as from 0.002, such as from 0.003, such as from 0.004, such as from 0.005, such as from 0.006, such as from 0.007, such as from 0.008, such as from 0.009, such as from 0.01 , such as from 0.02, such as from 0.03, such as from 0.04, such as from 0.05, such as from 0.06, such as from 0.07, such as from 0.08, such as from 0.09, such as from 0.1 , such as from 0.2, such as from 0.3, such as from 0.4, such as from 0.5, such as from 0.6, such as from 0.7, such as from 0.8, such as from 0.9 % (w/v) non-ionic detergent to 0.002, such as to 0.003, such as to 0.004, such as to 0.005, such as to 0.006, such as to 0.007, such as to 0.008, such as to 0.009, such as to 0.01 , such as to 0.02, such as to 0.03, such as to 0.04, such as to 0.05, such as to 0.06, such as to 0.07, such as to 0.08, such as to 0.09, such as to 0.1 , such as to 0.2, such as to 0.3, such as to 0.4, such as to 0.5, such as to 0.6, such as to 0.7, such as to 0.8, such as to 0.9, such as to 1 % (w/v) of non-ionic detergents.

In one embodiment the non-ionic detergent is selected from polysorbate 80, polysorbate

20 and poloxamer.

In one embodiment the composition of the present invention comprises up to 0.01 , preferably up to 0.001 , more preferably up to 0.0001 % (w/v) of non-ionic detergents such as but not limited to polysorbate 80, polysorbate 20 and poloxamer.

In one embodiment, the composition according to the present invention is essentially detergent free.

Fatty acids

Typically the composition of the present invention comprises less than or equal to 25 mM fatty acids. The fatty acid can be any fatty acid such as saturated or unsaturated fatty acids as well as salts thereof. Preferably the fatty acid is one or more saturated fatty acids such as a fatty acid selected from the group consisting of propanoic acid, butanoic acid, pentanoic acid, hexanoic acid, heptanoic acid, octanoic acid, nonanoic acid, decanoic acid, undecanoic acid, dodecanoic acid, tridecanoic acid, tetradecanoic acid, pentadecanoic acid, hexadecanoic acid, heptadecanoic acid, octadecanoic acid, nonadecanoic acid, eicosanoic acid, heneicosanoic acid, docosanoic acid, tricosanoic acid, tetracosanoic acid, pentacosanoic acid, hexacosanoic acid, heptacosanoic acid, octacosanoic acid, nonacosanoic acid, triacontanoic acid, henatriacontanoic acid, dotriacontanoic acid, tritriacontanoic acid, tetratriacontanoic acid, pentatriacontanoic acid and hexatriacontanoic acid. In one embodiment the composition comprises less than or equal to 25 mM fatty acids, such as less than or equal to 20 mM fatty acids, such as less than or equal to 15 mM fatty acids, such as less than or equal to 10 mM fatty acids, such as less than or equal to 5 mM fatty acids, such as less than or equal to 2 mM fatty acids, such as less than or equal to 1 mM fatty acids.

In one embodiment the composition comprises less than 25 mM fatty acids, such as less than 20 mM fatty acids, such as less than 15 mM fatty acids, such as less than 15 mM fatty acids, such as less than 14 mM fatty acids, such as less than 13 mM fatty acids, such as less than 12 mM fatty acids, such as less than 1 1 mM fatty acids, such as less than 10 mM fatty acids, such as less than 9 mM fatty acids, such as less than 8 mM fatty acids, such as less than 7 mM fatty acids, such as less than 6 mM fatty acids, such as less than 5 mM fatty acids, such as less than 4 mM fatty acids, such as less than 3 mM fatty acids, such as less than 2 mM fatty acids, such as less than 1 mM fatty acids, such as less than 0.5 mM fatty acids, such as less than 0.1 mM fatty acids, such as less than 0.05 mM fatty acids, such as less than 0.01 mM fatty acids, such as wherein the composition is essentially free of fatty acids.

In a preferred embodiment the fatty acid is octanoate (octanoic acid). In one embodiment the composition comprises less than or equal to 25 mM octanoate, such as less than or equal to 20 mM octanoate, such as less than or equal to 15 mM octanoate, such as less than or equal to 10 mM octanoate, such as less than or equal to 5 mM octanoate, such as less than or equal to 2 mM octanoate, such as less than or equal to 1 mM octanoate.

In one embodiment the composition comprises less than 25 mM octanoate, such as less than 20 mM octanoate, such as less than 15 mM octanoate, such as less than 15 mM octanoate, such as less than 14 mM octanoate, such as less than 13 mM octanoate, such as less than 12 mM octanoate, such as less than 1 1 mM octanoate, such as less than 10 mM octanoate, such as less than 9 mM octanoate, such as less than 8 mM octanoate, such as less than 7 mM octanoate, such as less than 6 mM octanoate, such as less than 5 mM octanoate, such as less than 4 mM octanoate, such as less than 3 mM octanoate, such as less than 2 mM octanoate, such as less than 1 mM octanoate, such as less than 0.5 mM octanoate, such as less than 0.1 mM octanoate, such as less than 0.05 mM octanoate, such as less than 0.01 mM octanoate, such as wherein the composition is essentially free of octanoate.

For example, may comprise less than or equal to 25 mM octanoate or total fatty acids is less than or equal to 20 mM octanoate or total fatty acids, such as less than or equal to 15 mM octanoate or total fatty acids, such as less than or equal to 10 mM octanoate or total fatty acids, such as less than or equal to 5 mM octanoate or total fatty acids, such as less than or equal to 2.28 mM octanoate or total fatty acids, such as less than or equal to 2.16 mM octanoate or total fatty acids, such as less than or equal to 2 mM octanoate or total fatty acids, such as less than or equal to 1.52 mM octanoate or total fatty acids, such as less than or equal to 1.44 mM octanoate or total fatty acids, such as less than or equal to 1.2 mM octanoate or total fatty acids, such as less than or equal to 1 mM octanoate or total fatty acids, such as less than or equal to 800 uM octanoate or total fatty acids, such as less than or equal to 720 uM octanoate or total fatty acids, such as less than or equal to 456 uM octanoate or total fatty acids, such as less than or equal to 400 mM octanoate or total fatty acids, such as less than or equal to 304 mM octanoate or total fatty acids, such as less than or equal to 288 mM octanoate or total fatty acids, such as less than or equal to 240 uM octanoate or total fatty acids, such as less than or equal to 160 uM octanoate or total fatty acids, such as less than or equal to 152 uM octanoate or total fatty acids, such as less than or equal to 144 uM octanoate or total fatty acids, such as less than or equal to 80 uM octanoate or total fatty acids.

The composition may comprise a molar ratio of octanoate to albumin that is less than or equal to 20:1 , 19:1 , 18:1 , 17:1 , 16:1 , 15:1 , 14:1 , 13:1 , 12:1 , 1 1 :1 , 10:1 , 9:1 , 8:1 , 7:1 , 6:1 , 5:1 , 4:1 , 3: 1 , 2:1 or 1 :1. A molar ratio of less than or equal to 16:1 , 1 1 :1 or 5:1 is preferred.

In one embodiment the composition comprises less than 25 mM hydrophobic molecules e.g. phospholipids, such as less than 20 mM hydrophobic molecules e.g. phospholipids, such as less than 15 mM hydrophobic molecules e.g. phospholipids, such as less than 15 mM hydrophobic molecules e.g. phospholipids, such as less than 14 mM hydrophobic molecules e.g. phospholipids, such as less than 13 mM hydrophobic molecules e.g. phospholipids, such as less than 12 mM hydrophobic molecules e.g. phospholipids, such as less than 1 1 mM hydrophobic molecules e.g. phospholipids, such as less than 10 mM hydrophobic molecules e.g. phospholipids, such as less than 9 mM hydrophobic molecules e.g. phospholipids, such as less than 8 mM hydrophobic molecules e.g. phospholipids, such as less than 7 mM hydrophobic molecules e.g. phospholipids, such as less than 6 mM hydrophobic molecules e.g. phospholipids, such as less than 5 mM hydrophobic molecules e.g. phospholipids, such as less than 4 mM hydrophobic molecules e.g. phospholipids, such as less than 3 mM hydrophobic molecules e.g. phospholipids, such as less than 2 mM hydrophobic molecules e.g. phospholipids, such as less than 1 mM hydrophobic molecules e.g. phospholipids, such as less than 0.5 mM hydrophobic molecules e.g. phospholipids, such as less than 0.1 mM hydrophobic molecules e.g. phospholipids, such as less than 0.05 mM hydrophobic molecules e.g. phospholipids, such as less than 0.01 mM hydrophobic molecules e.g. phospholipids, such as wherein the composition is essentially free of hydrophobic molecules e.g. phospholipids.

In one embodiment the term hydrophobic molecules includes fatty acids such as octanoate, but excludes detergents such as non-ionic detergents, such as polysorbate 80.

The composition of the present invention may include or exclude various amphiphilic compounds. Thus the present invention may include one type of amphiphilic compound but exclude another. Alternatively the composition may exclude essentially all amphiphilic compounds such as detergents, fatty acids and/or phospholipids.

In one embodiment the composition according to the invention comprises less than or equal to 25 mM amphiphilic compounds. In another embodiment the composition according to the invention is essentially free from amphiphilic compounds.

Free amino acids

The composition of the present invention may include one or more free amino acids, including natural amino acids selected from the group consisting of tryptophan, phenylalanine, tyrosine, glycine, alanine, valine, leucine, isoleucine, methionine, proline, serine, threonine, cysteine, asparagine, glutamine, aspartate, glutamate, lysine, arginine and histidine, or modified and non-natural amino acids. Any one of the amino acids of the composition of the present invention may be either an L-amino acid or a D-amino acid.

The composition according to one embodiment of the present invention typically comprises at least 5 mg/ml, at least 10 mg/ml, at least 15 mg/ml or at least 20 mg/ml of one or more (several) free amino acids such as phenylalanine, tyrosine, glycine, alanine, valine, leucine, isoleucine, methionine, proline, serine, threonine, cysteine, asparagine, glutamine, aspartate, glutamate, lysine, arginine, histidine, or modified and non-natural amino acids. Glycine and/or arginine are preferred. For example, the composition may comprise from 5, 10, 15, 20, 25, 50, 75 to 10, 15, 20, 25, 50, 75 mg/ml of the above mentioned amino acid(s). For example, the composition may comprise from about 25, 50, 75, 100, 125, 150, 175 to about 50, 75, 100, 125, 150, 175 mM of one or more amino acids.

The composition according to another embodiment of the present invention typically comprises less than 5 mM free amino acids such as less than 4 mM free amino acids, such as less than 3 mM free amino acids, such as less than 2 mM free amino acids, such as less than 1 mM free amino acids, such as less than 0.5, such as less than 0.1 , such as less than 0.01 , such as less than 0.005, such as less than 0.001 mM free amino acids or essentially no free amino acids.

In one embodiment the composition is essentially free from free amino acids.

The composition may or may not comprise free tryptophan or N-acetyl-tryptophan. In a preferred embodiment the composition comprises free tryptophan or N-acetyl-tryptophan.

In one embodiment the composition comprises less than 5, such as less than 4, such as less than 3, such as less than 2, such as less than 1 , such as less than 0.5, such as less than 0.1 , such as less than 0.01 , such as less than 0.005, such as less than 0.001 mM tryptophan or N-acetyl tryptophan, or essentially no tryptophan or N-acetyl tryptophan. Salt

The composition according to the present invention may include any suitable salt such as but not limited to bromide, chloride, fluoride, hydride, iodide, nitride, oxide, phosphide, sulfide, peroxide, borate, bromate, hypobromite, carbonate, hydrogen carbonate, bicarbonate, chlorate, perchlorate, chlorite, hypochlorite, chromate, iodate, nitrate, nitrite, phosphate, hydrogen phosphate, dihydrogen phosphate, phosphite, sulfate, thiosulfate, hydrogen sulfate, bisulfate, sulfite, hydrogen sulfite, bisulfite, acetate, formate, oxalate, hydrogen oxalate, bioxalate, hydrogen sulfide, bisulfide, telluride, amide, thiocyanate, muriate (HCI), succinate and maleate salts or any combination thereof. Alternatively salts of the composition of the present invention may also include derivatives from nontoxic inorganic acids such as hydrochloric, nitric, phosphoric, sulphuric, hydrobromic, hydriodic, hydrofluoric, phosphorous and the like, as well as the salts derived from nontoxic organic acids, such as aliphatic mono and dicarboxylic acids, phenyl-substituted alkanoic acids, hydroxy alkanoic acids, alkanedioic acids, aromatic acids, aliphatic and aromatic sulfonic acids, etc. Such salts thus include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogenphosphate, dihydrogenphosphate, metaphosphate, pyrophosphate, chloride, bromide, iodide, acetate, trifluoroacetate, propionate, caprylate, isobutyrate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, mandelate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, phthalate, benzenesulfonate, toluenesulfonate, phenylacetate, citrate, lactate, tartrate, methanesulfonate, and the like.

In one embodiment the composition according to the present invention comprises less than 500 mM salt, preferably less than 400 mM salt, preferably less than 300 mM salt, preferably less than 250 mM salt, preferably less than 200 mM salt, preferably less than 150 mM salt, preferably less than 100 mM salt, preferably less than 50 mM salt, preferably less than 25 mM salt, preferably free of salt.

In one embodiment the composition according to the present invention comprises less than or equal to 300 mM salt.

In one embodiment the composition according to the present invention comprises less than or equal to 180 mM salt.

In one embodiment the composition comprises less than or equal to 300, 250, 200, 150,

100, 50, 25 mM salt, such as from 25, 50, 100, 150, 200, 250 to 50, 100, 150, 200, 250 or 300 mM salt.

Salts of the invention may include salts of metals, such as monovalent (e.g. Group 1 ) metals and divalent (e.g. Group 2 and transition element) metals, and salts of ammonium. Salts include NaCI, and KCI.

Metal ions

As demonstrated in the present example section, stability of a peptide, such as insulin, in a zinc ion free composition is enhanced in the presence of stripped albumin. Thus in one embodiment, the composition of the present invention is essentially free of zinc.

In a further embodiment, the composition according to the present invention is essentially metal ion free, such as essentially free from Zn²⁺, Ca²⁺, Mg²⁺, Mn²⁺, Fe²⁺, Cu²⁺, Co²⁺ and/or Ni²⁺ ions. Acid/Base considerations

The composition of the present invention preferably has a pH of between 4 and 9; such as between 4 and 8; such as between 4 and 7; such as between 5 and 8; such as between 6 and 8; preferably between 6.5 and 7.5 such as wherein said composition has a pH of about 7.

For insulin, or analog thereof, a pH of about 4 to about 7 such as about 7 is preferred.

For GLP-2, or analog thereof, a pH of at least about 6, such as from about 6 to 9, such as from about 6 to 8 is preferred.

For HIV fusion inhibitor, e.g. enfuvirtide, or analog thereof, a pH of at least 6 such as from about 6 to about 9 or from about 6 to about 8 is preferred.

For glucagon, a pH of from about 4.5 to about 5.5. such as about 5.0, is preferred.

Buffer

The composition may comprise a buffer such as a citrate buffer, a phosphate buffer or a histidine buffer. Phosphate buffer or histidine buffer are preferred. The buffer concentration may be from about 10 to about 150 mM, such as from about 30 to about 150 mM, such as from about 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 1 10, 120, 130, or 140 to about 20, 30, 40, 50, 60, 70, 80, 90, 100, 1 10, 120, 130, 140 or about 150 mM.

Stability

The stability of the composition comprising peptide and albumin may be more stable than a reference composition. The reference composition may be a composition which does not contain albumin. The reference composition may be a composition which contains more or less (preferably more) octanoate (or total fatty acid) and/or more or less (preferably more) polysorbate 80 (or total detergent) than the test composition. A reference composition may comprise 0.8 mM octanoate and/or total fatty acid. For example, lag time to reach a defined threshold (e.g. 5000, 10000 or 20000 RFU) in a ThT assay (e.g. Example 1 , 2 or 3a) may be at least 48, 60, 72, 96, or 120 hours or may be at least 48, 60, 72, 96, or 120 hours longer than the lag time for a reference composition or may be at least 2- fold, 3- fold, 4- fold, 5- fold, 6- fold, 7- fold, 8- fold, 9- fold or 10-fold longer than the lag time for a reference composition. For example, AUC in a SE-HPLC assay (e.g. Example 3b) may be at least 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, or 99% higher after 12, 24, 36, 48, 60, or 72 hours of the initial AUC of the test composition or of a reference composition, or may be at least 2-fold, 3-fold, 4-fold or 5-fold higher than the after 12, 24, 36, 48, 60, or 72 hours of the initial AUC of the test composition or of a reference composition. For example, the time taken for the initial peptide AUC value to reduce to 50% of that initial value may be at least 12, 24, 36, 48, 60, or 72 hours, or at least 12, 24, 36, 48, 60, or 72 hours longer for a test composition than for a reference composition or at least 2-fold, 3-fold, 4-fold or 5-fold longer for a test composition than for a reference composition. Uses of the composition

Insulin, and many other peptides, undergoes aggregation-coupled mis-folding to form a cross-β assembly. Such fibrillation has long complicated manufacture of insulin and its use in the therapy of diabetes mellitus. It is thus desirable to provide a composition which enhances stability i.e. reduces the rate of fibrillation of peptides. The present inventors have surprisingly demonstrated that the composition according to the present invention is suitable for preventing and/or reducing peptide fibrillation and thereby stabilizing the peptide composition.

The composition as defined herein above may thus be used for stabilizing peptides which under non-stabilizing conditions form peptide fibrils.

Accordingly, one aspect the invention concerns use of a highly purified albumin composition for preventing and/or reducing formation of peptide fibrils, such as insulin fibrils, e.g. in an essentially zinc ion free formulation.

In one embodiment, the composition according to the present invention is used for preventing and/or reducing formation of peptide fibrils.

In another embodiment, the invention concerns a method of stabilizing an essentially zinc ion free insulin composition comprising dissolving zinc ion free insulin in an aqueous solution of highly purified albumin, wherein the aqueous solution is essentially detergent free, such as free from non-ionic detergents such as free from polysorbate 80 or 20.

In another embodiment, the invention concerns a method of stabilizing an essentially zinc ion free insulin composition comprising dissolving zinc ion free insulin in an aqueous solution of highly purified albumin, wherein the aqueous solution is essentially free of octanoate and/or essentially free of fatty acids. In one aspect the invention concerns a method of preventing and/or reducing formation of peptide fibrils in an aqueous solution, wherein the method comprises dissolving the peptide in an aqueous solution of albumin, wherein the aqueous solution is essentially free of amphiphilic compounds, such as free from non-ionic detergents and/or free from octanoate and/or free from fatty acids.

In one embodiment of the above method, the composition comprises less than 25 mM fatty acids and is essentially free from non-ionic detergents.

In one aspect, the invention concerns a method of preventing and/or reducing formation of peptide fibrils, the method comprising preparing a composition as defined in any one of the embodiments herein.

The invention is further defined by the following paragraphs:

1. A composition comprising:

a. from 0.01 mg/ml to 300 mg/ml peptide (such as a peptide having a size of from 10 to 100 amino acids); b. from 0.01 mg/ml to 500 mg/ml albumin

2. A composition comprising:

a. from 0.01 mg/ml to 300 mg/ml peptide (such as a peptide having a size of from

10 to 100 amino acids);

b. from 0.01 mg/ml to 500 mg/ml albumin

wherein the composition comprises less than or equal to 25 mM fatty acid and/or less than 0.001 % (w/v) detergent.

3. The composition according to paragraph 1 or 2 comprising less than or equal to 25 mM octanoate and less than or equal to 0.001 % (w/v) polysorbate 80.

4. The composition according to paragraph 1 , 2 or 3 comprising less than or equal to 25 mM fatty acids.

5. The composition according to any preceding paragraph comprising less than or equal to 0.001 % (w/v) detergent.

6. The composition according to any preceding paragraph, wherein the peptide is a non- lipopeptide.

7. The composition according to any preceding paragraph comprising from 1 to 20 mg/ml peptide.

8. The composition according any preceding paragraph comprising from 1 to 20 mg/ml albumin.

9. The composition according to any preceding paragraph, wherein the peptide is essentially in non-fibril form.

10. The composition according to any preceding paragraph, wherein the peptide comprises two or more different peptide chains and/or the composition comprises two or more different peptides.

1 1. The composition according to any preceding paragraph, wherein the peptide is selected from the group consisting of insulin, insulin analogs, glucagon or analog thereof, GLP-1 or analog thereof, GLP-2 or analog thereof, HIV fusion inhibitor (e.g. enfurvitide) or a fragment or variant thereof.

12. The composition according to any preceding paragraph, wherein the peptide comprises less than 200 amino acid residues, such as less than 195 amino acid residues, such as less than 190 amino acid residues, such as less than 185 amino acid residues, such as less than 180 amino acid residues, such as less than 175 amino acid residues, such as less than 160 amino acid residues, such as less than 150 amino acid residues, such as less than 140 amino acid residues, such as less than 130 amino acid residues, such as less than 125 amino acid residues, such as less than 120 amino acid residues, such as less than 1 15 amino acid residues, such as less than 1 10 amino acid residues, such as less than 105 amino acid residues, such as less than 100 amino acid residues, such as less than 95 amino acid residues, such as less than 90 amino acid residues, such as less than 85 amino acid residues, such as less than 80 amino acid residues, such as less than 75 amino acid residues, such as less than 70 amino acid residues, such as less than 65 amino acid residues, such as less than 60 amino acid residues, such as less than 55 amino acid residues, such as less than 50 amino acid residues, such as less than 45 amino acid residues, such as less than 40 amino acid residues, such as less than 35 amino acid residues, such as less than 30 amino acid residues, such as less than 25 amino acid residues, such as less than 20 amino acid residues, such as less than 15 amino acid residues.

13. The composition according to any preceding paragraph, wherein the less than 0.001 % (w/v) detergent is less than 0.001 % (w/v) non-ionic detergent.

14. The composition according to any preceding paragraph, wherein the less than 0.001 % (w/v) detergent is less than 0.001 % (w/v) anionic detergent.

15. The composition according to any preceding paragraphs wherein the less than 0.001 % (w/v) detergent is less than 0.001 % (w/v) cationic detergent.

16. The composition according to any preceding paragraphs wherein the less than 0.001 % (w/v) detergent is less than 0.001 % (w/v) zwitterionic detergent.

17. The composition according to any of paragraphs 13 to 16, wherein the less than 0.001 % (w/v) non-ionic detergent is less than 0.001 % (w/v) polysorbate 80.

18. The composition according to any of paragraphs 13 to 16, wherein the less than 0.001 % (w/v) non-ionic detergent is less than 0.001 % (w/v) polysorbate 20.

19. The composition according to any of paragraphs 13 to 16, wherein the less than 0.001 % (w/v) non-ionic detergent is less than 0.001 % (w/v) poloxamer.

20. The composition according to any preceding paragraph, comprising less than or equal to 3.325^*10^"4 % (w/v) polysorbate 80 or 20, such as less than or equal to 2.85^*10^"4 % (w/v) polysorbate 80 or 20, such as less than or equal to 2.375^*10^"4 % (w/v) polysorbate 80 or 20, such as less than or equal to 1.425^*10^"4 % (w/v) polysorbate 80 or 20, such as less than or equal to 9^*10^"5 % (w/v) polysorbate 80 or 20, such as less than or equal to 6.625^*10^"5 % (w/v) polysorbate 80 or 20, such as less than or equal to 5.7^*10^"5 % (w/v) polysorbate 80 or 20, such as less than or equal to 5^*10^"5 % (w/v) polysorbate 80 or 20, such as less than or equal to 4.75^*10^"5 % (w/v) polysorbate 80 or 20, such as less than or equal to 4.5^*10^"5 % (w/v) polysorbate 80 or 20, such as less than or equal to 4.75^*10^"5 % (w/v) polysorbate 80 or 20, such as less than or equal to 2.85^*10^"5 % (w/v) polysorbate 80 or 20, such as less than or equal to 2.5^*10^"5 % (w/v) polysorbate 80 or 20, such as less than or equal to 1.9^*10^"5 % (w/v) polysorbate 80 or 20, such as less than or equal to 1.8^*10^"5 % (w/v) polysorbate 80 or 20, such as less than or equal to 1 ^* 10^"5 % (w/v) polysorbate 80 or 20, such as less than or equal to 9.5^*10^"6 % (w/v) polysorbate 80 or 20, such as less than or equal to 9^*10^"6 % (w/v) polysorbate 80 or 20, such as less than or equal to 5^*10^"6 % (w/v) polysorbate 80 or 20, most preferably substantially free of polysorbate 80 or 20.

21. The composition according to any preceding paragraph, wherein the composition is essentially detergent free.

22. The composition according to any preceding paragraph, wherein the less than or equal to 25 mM fatty acids is less than or equal to 25 mM octanoate.

23. The composition according to any preceding paragraph, wherein the less than or equal to 25 mM octanoate or total fatty acids is less than or equal to 20 mM octanoate or total fatty acids, such as less than or equal to 15 mM octanoate or total fatty acids, such as less than or equal to 10 mM octanoate or total fatty acids, such as less than or equal to 5 mM octanoate or total fatty acids, such as less than or equal to 2.28 mM octanoate or total fatty acids, such as less than or equal to 2.16 mM octanoate or total fatty acids, such as less than or equal to 2 mM octanoate or total fatty acids, such as less than or equal to 1 .52 mM octanoate or total fatty acids, such as less than or equal to 1.44 mM octanoate or total fatty acids, such as less than or equal to 1 .2 mM octanoate or total fatty acids, such as less than or equal to 1 mM octanoate or total fatty acids, such as less than or equal to 800 uM octanoate or total fatty acids, such as less than or equal to 720 uM octanoate or total fatty acids, such as less than or equal to 456 uM octanoate or total fatty acids, such as less than or equal to 400 mM octanoate or total fatty acids, such as less than or equal to 304 mM octanoate or total fatty acids, such as less than or equal to 288 mM octanoate or total fatty acids, such as less than or equal to 240 uM octanoate or total fatty acids, such as less than or equal to 160 uM octanoate or total fatty acids, such as less than or equal to 152 uM octanoate or total fatty acids, such as less than or equal to 144 uM octanoate or total fatty acids, such as less than or equal to 80 uM octanoate or total fatty acids.

24. The composition according to any preceding paragraph, wherein the composition is essentially octanoate free or substantially free of total fatty acids.

25. The composition according to any preceding paragraph, wherein the composition comprises free glycine or free arginine.

26. The composition according to paragraph 25 wherein the concentration of free glycine or free or free arginine is at least 5 mg/ml, such as at least 10 mg/ml, such as at least 20 mg/ml.

27. The composition according to paragraph 25 or 26 wherein the concentration of free glycine or free arginine is less than or equal to 100 mg/ml, such as less than or equal to 80 mg/ml, such as less than or equal to 60 mg/ml, such as less than or equal to 50 mg/ml, such as less than or equal to 40 mg/ml, such as less than or equal to 30 mg/ml.

28. The composition according to any preceding paragraph, wherein the composition comprises less than or equal to 25 mM small hydrophobic molecules. 29. The composition according to any preceding paragraph, wherein the composition comprises less than or equal to 25 mM amphiphilic compounds.

30. The composition according to any preceding paragraph, wherein the composition is essentially free from amphiphilic compounds.

31. The composition according to any preceding paragraph, wherein the composition comprises less than or equal to 5 mM free N-acetyl tryptophan or free tryptophan or less than or equal to 5 mM free amino acids.

32. The composition according to paragraph 31, wherein the less than 5 mM free N-acetyl tryptophan, free tryptophan or free amino acids is less than 4 mM free N-acetyl tryptophan, free tryptophan or free amino acids, such as less than 3 mM free N-acetyl tryptophan, free tryptophan or free amino acids, such as less than 2 mM free N-acetyl tryptophan, free tryptophan or free amino acids, such as less than 1 mM free N-acetyl tryptophan, free tryptophan or free amino acids.

33. The composition according to paragraph 31 or 32, wherein the free amino acid is selected from the group consisting of tryptophan, N-acetyl-tryptophan, phenylalanine, tyrosine, glycine, alanine, valine, leucine, isoleucine, methionine, proline, serine, threonine, cysteine, asparagine, glutamine, aspartate, glutamate, lysine, arginine and histidine.

34. The composition according to any preceding paragraph, wherein the composition comprises less than 500 mM salt, preferably less than 400 mM salt, preferably less than

300 mM salt, preferably less than 250 mM salt, preferably less than 200 mM salt, preferably less than 150 mM salt, preferably less than 100 mM salt, preferably less than 50 mM salt, preferably less than 25 mM salt, preferably free of salt.

35. The composition according to any preceding paragraph, wherein the composition comprises less than or equal to 300 mM salt.

36. The composition according to any preceding paragraph, wherein the composition comprises less than or equal to 180 mM salt.

37. The composition according to any preceding paragraph, wherein the molar ratio of peptide to albumin is from 1 part peptide to 2000 parts albumin (1: 2000) to 3000 parts peptide to 1 part albumin (3000: 1) such as from about 1:1000, 1:500, 1:250, 1:100,

1:80, 1:50, 1:40, 1:36, 1:25, 1:20, 1:10, 1:7, 1:5.2, 1:5, 1:4.3, 1:4, 1:3.7, 1:3.3, 1.3, 1:2, 1:1.15, 1:1, 1.15:1, 1.4:1, 1.5:1, 2:1, 3:1, 4:1, 5:1, 10: 1, 50:1, 100:1 , 200:1 , 250:1 , 500:1, 750:1, 1000:1, or 1500:1 to about 1:500, 1:250, 1:100, 1:80, 1:50, 1:40, 1:36, 1:25, 1:20, 1:10, 1:7, 1:5.2, 1:5, 1:4.3, 1:4, 1:3.7, 1:3.3, 1.3, 1:2, 1:1.15, 1:1, 1.15:1, 1.4:1, 1.5:1, 2:1, 3:1, 4:1, 5:1, or 10:1, 50:1, 100:1, 200:1 , 250:1 , 500:1 , 750:1 , 1000:1, 1500:1 or 2500:1.

38. The composition according to any preceding paragraph, wherein the molar ratio of octanoate to albumin is less than or equal to 20:1, 19:1, 18:1, 17:1, 16:1, 15:1, 14:1, 13:1 , 12: 1 , 1 1 :1 , 10:1 , 9:1 , 8:1 , 7:1 , 6:1 , 5:1 , 4:1 , 3: 1 , 2:1 or 1 : 1 , preferably less than or equal to 16:1 , 1 1 :1 or 5:1.

39. The composition according to any preceding paragraph, wherein the composition is essentially free of zinc.

40. A composition which is essentially free of zinc comprising:

a. from 0.01 mg/ml to 300 mg/ml insulin or insulin analog;

b. from 0.01 mg/ml to 500 mg/ml albumin

41. The composition according to any preceding paragraph, wherein the composition is essentially metal ion free.

42. The composition according to paragraph 41 , wherein the essentially metal ion free composition is essentially free of Zn²⁺, Ca²⁺, Mg²⁺, Mn²⁺, Fe²⁺, Cu²\ Co²⁺ and/or Ni²⁺ ions.

43. The composition according to any preceding paragraph, comprising from 0.01 to 500 mg/ml albumin and from 1 to 100 mg/ml peptide.

44. The composition according to any preceding paragraph, comprising from 1 to 250 mg/ml albumin and from 1 to 100 mg/ml peptide.

45. The composition according to any preceding paragraph, comprising from 1 to 100 mg/ml albumin and from 1 to 100 mg/ml peptide.

46. The composition according to any preceding paragraph, comprising from 1 to 30 mg/ml albumin and from 0.01 to 100 mg/ml peptide.

47. The composition according to any preceding paragraph wherein said composition has a pH of between 4 and 9; such as between 4 and 8; such as between 4 and 7; such as between 5 and 8; such as between 6 and 8; preferably between 6.5 and 7.5 such as wherein said composition has a pH of about 7.

48. The composition according to paragraph 47 wherein the peptide is selected from insulin or an insulin analog and the pH is from about 4 to about 7 (preferably about 7).

49. The composition according to paragraph 48 wherein the molar ratio of insulin to albumin is from about 10:1 to about 1 :10, such as from about 5:1 to about 1 :5, such as about

3.3:1.

50. The composition according to paragraph 48 or 49 wherein the albumin concentration is from about 0.5 to about 20 mg/ml (such as from about 1 to about 19, such as from about 3.7 to about 18.5 mg/ml, such as from about 1.5 to about 12, such as from about 1 .7 to about 10) and the insulin concentration is from about 0.01 to about 1 mg/ml (such as from about 0.1 to about 1 , such as from such as from about 0.3 to about 0.8 mg/ml, such as from about 0.33 to about 0.73). The composition according to paragraph 48, 49 or 50 wherein the molar ratio of octanoate to albumin is from about 1 :1 to about 20:1 such as from about 5:1 to about 16:1 .

The composition according to any of paragraphs 48 to 51 wherein the insulin analog is lispro.

The composition according to paragraph 47 wherein the peptide is selected from GLP-2 or GLP-2 analog and the pH is at least about 6, such as at least about 7, such as at least about 8, such as from about 6 to about 8.

The composition according to paragraph 53 wherein the molar ratio of GLP-2 or GLP-2 analog to albumin is from about 10:1 to about 1 :10, such as from about 5: 1 to about 1 :5, such as from about 5:1 to about 1 :2, such as from about 5: 1 to about 1 :1.4.

The composition according to paragraph 52 or 53 wherein the albumin concentration is from about 0.5 to about 20 mg/ml (such as from about 1 to about 10, such as from about 1 to about 5 mg/mlsuch as from about 0.1 to about 2 mg/ml, such as from about 0.2 to about 1.8 mg/ml, such as from about 0.21 to about 1 .74 mg/ml) and the GLP-2 or GLP-2 analog concentration is from about 0.1 to about 5 mg/ml (such as from about 0.21 to about 2.4 mg/ml).

The composition according to any of paragraphs 52, 53 or 54 wherein the molar ratio of octanoate to albumin is from about 1 :1 to about 20:1 such as from about 5:1 to about 16:1 .

The composition according to any of paragraphs 52 to 56 wherein the GLP-2 analog is teduglutide.

The composition according to paragraph 47 wherein the peptide is a GLP-1 analog. The composition according to paragraph 58 wherein the molar ratio of GLP-1 analog to albumin is from about 10:1 to about 1 :10, such as from about 7:1 to about 1 :7, such as from about 4:1 to about 6:1 , such as about 5.2:1 :1.

The composition according to paragraph 58 or 59 wherein the albumin concentration is from about 0.5 to about 20 mg/ml (such as from about 1 to about 10, such as from about 0.5 to about 7, such as from about 1 to about 5 mg/ml) and the GLP-1 analog concentration is from about 0.2 to about 0.3 mg/ml (such as about 0.26 mg/ml).

The composition according to paragraph 58, 59 or 60 wherein the molar ratio of octanoate to albumin is from about 1 :1 to about 20:1 such as from about 5:1 to about 16:1 .

The composition according to any of paragraphs 58 to 61 wherein the GLP-1 analog is liraglutide.

The composition according to paragraph 47 wherein the peptide is an HIV inhibitor such as enfuvirtide or analog thereof and the pH is at least about 6, such as at least about 7, such as at least about 8, such as from about 6 to about 8. The composition according to paragraph 63 wherein the molar ratio of enfuvirtide or analog thereof to albumin is from about 50:1 to about 1 :50, such as from about 45: 1 to about 7:1 , such as from about 39:1 to about 41 :1 or about 6:1 to about 8:1 , such as about 40:1 or about 8:1.

The composition according to paragraph 63 or 64 wherein the albumin concentration is from about 0.5 to about 20 mg/ml (such as from about 1 to about 10, such as about 1 to about 5 mg/ml) and the enfuvirtide or analog thereof concentration is from from about 1 to about 50 mg/ml (such as from about 4 to about 45 mg/ml, such as from about 48 to about 45 mg/ml, such as from about 38 to about 45 mg/ml, such as from about 39.9 to about 42.7 mg/ml).

The composition according to paragraph 63, 64 or 65 wherein the molar ratio of octanoate to albumin is from about 1 :1 to about 20:1 such as from about 5:1 to about 16:1 , e.g. about 8:1.

The composition according to paragraph 47 wherein the peptide is glucagon and the pH is from about 4.5 to about 5.5, preferably about 4.5 to about 5.5, most preferably about 5.0.

The composition according to paragraph 67, comprising arginine or glycine.

The composition according to any preceding paragraph wherein said composition consists essentially of albumin and insulin.

The composition according to any preceding paragraph wherein:

a. time taken for the composition to reach a threshold (e.g. 5000, 10000 or 20000 relative fluorescence units) when analyzed by ThT assay is at least twice the time taken for a reference composition to reach the same threshold);

b. time taken for the composition to reach a threshold (e.g. 5000, 10000 or 20000 relative fluorescence units) when analyzed by ThT assay is at least 48, 60, 72, 96, or 120 hours;

c. time taken for the composition to reach a threshold (e.g. 5000, 10000 or 20000 relative fluorescence units) when analyzed by ThT assay is at least 48, 60, 72, 96, or 120 hours longer than the lag time for a reference composition and/or at least 2- fold, 3- fold, 4- fold, 5- fold, 6- fold, 7- fold, 8- fold, 9- fold or 10-fold longer than the lag time for a reference composition;

d. AUC in a SE-HPLC assay (e.g. Example 3b) is at least 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, or 99% higher after 12, 24, 36, 48, 60, or 72 hours of the initial AUC of the test composition or of a reference composition, or may be at least 2-fold, 3-fold, 4-fold or 5-fold higher after 12, 24, 36, 48, 60, or 72 hours of the initial AUC of the test composition or of a reference composition; and/or e. Time taken for the initial AUC value of the test composition to reduce to 50% of that initial value may be at least 12, 24, 36, 48, 60, or 72 hours, or at least 12, 24, 36, 48, 60, or 72 hours longer for a test composition than for a reference composition or at least 2-fold, 3-fold, 4-fold or 5-fold longer for a test composition than for a reference composition.

71. The composition according to any preceding paragraph comprising a buffer selected from citrate buffer, phosphate buffer or histidine; preferably phosphate buffer or histidine, 72. The composition according to paragraph 71 wherein the citrate buffer is from about 50 to about 150 mM, preferably about 80 to about 120 mM, most preferably about 100 mM. 73. The composition according to paragraph 71 wherein the phosphate buffer is from about 50 to about 150 mM, preferably about 80 to about 120 mM, most preferably about 100 mM.

74. The composition according to paragraph 71 wherein the histidine buffer is from about 10 to about 100 mM, preferably from about 30 to about 50 mM, most preferably about 40 mM.

75. Use of highly purified albumin composition for preventing and/or reducing formation of insulin fibrils in an essentially zinc ion free formulation.

76. Use of the composition according to any one of paragraphs 1 to 75 for preventing and/or reducing formation of peptide fibrils.

77. A method of stabilizing an essentially zinc ion free insulin composition comprising dissolving zinc ion free insulin in an aqueous solution of highly purified albumin, wherein the aqueous solution is essentially detergent free, essentially octanoate free, essentially polysorbate 80 free, essentially fatty acid free, essentially detergent free and/or essentially amphiphilic compound free.

78. A method of preventing and/or reducing formation of peptide fibrils in an aqueous solution, the method comprising dissolving the peptide in an aqueous solution of albumin, wherein the aqueous solution is essentially free of octanoate, essentially free of polysorbate 80, essentially free of fatty acids, essentially free of detergents and/or amphiphilic compounds.

79. A method of preventing and/or reducing formation of peptide fibrils, the method comprising preparing a composition as defined in any of paragraphs 1 to 75. The present invention is further described by the following examples that should not be construed as limiting the scope of the invention.

Examples

Example 1 : Sample preparation

Preparation of Stripped rAlbumin samples: A 10 % solution of Albucult^® was diluted to

5% (w/v) with de-ionized water and dialyzed three times against de-ionized water using a 20K dialysis membrane. 0.75 gram activated charcoal per gram protein was added. (The amount of charcoal is dependent on the lipid content of the sample). The pH was subsequently lowered to 3 by addition of 0.5 M HCI and the solution was mixed for 4 hours at 5^°C. The charcoal was removed by filtering through a 0.2 μπι (0.2 micron) filter and the pH was adjusted to 7 by addition of 0.5 M NaOH. The stripped rAlbumin was concentrated using Millipore UFC tubes in PDVF to a concentration of 96 mg/ml.

Preparation of human insulin samples for Example 2: Human insulin was obtained from Sigma and contains Zn²⁺ in a molar ratio of 2-3:6 (Zn²⁺:insulin). Zn-insulin powder was wetted with water in a volume/weight based ratio of water to insulin of 5:2 and the pH was lowered by addition of 0.2 M HCI in a volume/weight based ratio of HCI to insulin of 5:2. When the protein was solubilized, the rest of the buffer components were added and the pH adjusted using 0.1 M NaOH. Human insulin samples without Zn were prepared by addition of 10 mM EDTA.

Preparation of insulin and insulin analog samples for Examples 4 to 8:

Preparation of human insulin samples: Human insulin was purchased from Novo Nordisk (Actrapid) (A chain: SEQ ID NO: 3, B chain: SEQ ID NO: 4). EDTA was added to the human insulin solution, resulting in a final EDTA concentration of 1 mM. Human insulin was loaded onto a C-18 column (similar to Daiso SP-120-15-ODS-AP) pre-equilibrated with 0.1 % formic acid and 10% ethanol at a flow rate of 7.6 column volume/h. Human insulin was eluted with a 10%-50% ethanol gradient and 0.1 % formic acid over two column volumes. The purified human insulin was vacuum dried on a Speed Vac system and dissolved in 2 mM acetic acid prior to use.

Preparation of insulin lispro samples: Insulin lispro was purchased from Eli Lilly (Humalog) (A chain: SEQ ID NO: 25, B chain: SEQ I D NO: 26). EDTA was added to the insulin lispro solution, resulting in a final EDTA concentration of 1 mM. Insulin lispro was loaded onto a C-18 column (similar to Daiso SP-120-15-ODS-AP) pre-equilibrated with 0.1 % formic acid and 10% ethanol at a flow rate of 7.6 column volume/h. Insulin lispro was eluted with a 10%-50% ethanol gradient and 0.1 % formic acid over two column volumes. The purified insulin lispro was vacuum dried on a Speed Vac system and dissolved in 2 mM acetic acid prior to use.

Preparation of liraglutide samples: Liraglutide was purchased from Novo Nordisk (Victoza) (SEQ ID NO: 27). EDTA was added to the liraglutide solution, resulting in a final EDTA concentration of 1 mM, and pH was adjusted to 7. Liraglutide was loaded onto a C-4 column (Daiso SP-300-15-C4-BIO) pre-equilibrated with 0.1 % formic acid and 5% ethanol at a flow rate of 8 column volume/h. Insulin lispro was eluted with a 5%-50% ethanol gradient and 0.1% formic acid over 0.5 column volumes. The purified liraglutide was vacuum dried on a Speed Vac system and dissolved in phosphate buffer pH 7.4 prior to use.

Preparation of Teduglutide samples: lyophilized teduglutide (SEQ ID NO: 28) was reconstituted in buffer as described in the examples below.

Preparation of Enfuvirtide samples: enfuvirtide (SEQ ID NO: 29), in powder form, was reconstituted in buffer as described in the examples below.

Determination of protein concentration: The concentration of stripped rAlbumin and Albucult was determined using the extinction coefficient at 279 nm (A279 = 0.531 cm^"1 for a 1 mg/ml) and a molar mass of rAlbumin of 66640 g/mol. The concentration of insulin, Zn insulin and insulin lispro was determined using the extinction coefficient at 276 nm (A276 = 1.05 cm^"1 for a 1 mg/ml, Porter, 1953) and a molar mass of human insulin of 5808 g/mol.

The concentration of liraglutide was determined using the extinction coefficient at 280 nm (A280 = 2.07 cm^"1 for a 1 mg/ml, Gasteiger, 2005) and a molar mass of liraglutide of 3383.7 g/mol.

The concentration of teduglutide was determined using the extinction coefficient at 280 nm (A280 = 1.47 cm^"1 for a 1 mg/ml, Gasteiger, 2005) and a molar mass of teduglutide of 3752.1 g/mol.

The concentration of enfuvirtide was determined using the extinction coefficient at 280 nm (A280 = 4.04 cm^"1 for a 1 mg/ml, Gasteiger, 2005) and a molar mass of enfuvirtide of 4450.8 g/mol.

Example 2: Determination of protein stability

ThT is a fluorescent probe that binds specifically to hydrophobic cavities running parallel to the fibril axis, e.g., between the protofilaments forming the fibrils and is used as a standard dye for amyloid detection. Upon binding to such fibrillar amyloids, a change in the excitation and emission maximum as well as a significantly enhanced fluorescence signal is observed for ThT (J Chem Biol. 2010 March; 3(1 ): 1-18; J Struct Biol. 2007 Sep;159(3):483-97).

ThT assays where all performed using a BMG Fluostar Omega fluorescence plate reader equipped with extinction and emission filters of 440±10 nm and 490±10 nm, respectively. Cycles were repeated with a cycle time of 600 seconds with 300 seconds orbital shaking at a temperature of 35°C in 96 well microtiter plates (Nunc 265301 ) covered with a transparent film (Film Nunc 233701 ). The assay was performed in 50 mM NaCI, 50 mM phosphate, 7.0 with 1 mM EDTA in all samples to immobilize the Zn²⁺ present and thereby favor the monomeric state of insulin. Each sample contained 1 mM ThT and 0.5 mg/ml (0.086 mM) insulin in a sample volume of 200 μΙ_. All results are presented as the mean of three measurements. The concentration Albumin when added was 5.74 mg/ml (0.086 mM) resulting in a molar ratio of 1 :1 insulin:Albumin

The data presented in Figure 1 show that the time dependent increase in fluorescence correlated with the formation of amyloid fibrils in the sample. No increase in the fluorescence is observed in the references of Albucult and stripped rAlbumin within the timeframe of the assay, whereas the fluorescence increases readily after approximately 20 hours in the human insulin reference. It is furthermore observed that the sample containing both human insulin and Albucult increases after approximately 5 hours whereas the sample containing human insulin formulated with stripped rAlbumin reveals no increase in the fluorescence within the timeframe of the assay.

The data presented in Figure 2 shows the time dependent increase in fluorescence correlated with the formation of amyloid fibrils in the sample. No increase in the fluorescence is observed in the references of Albucult and stripped rAlbumin within the timeframe of the assay, whereas the fluorescence increases readily after a little more than 10 hours in the human insulin reference. It is furthermore observed that the sample containing both human insulin and Albucult increases after approximately 3-5 hours whereas the sample containing human insulin formulated with stripped rAlbumin reveals a stronger resilience towards formation of amyloid fibrils which is not formed no increase in the fluorescence within the timeframe of the assay.

It is thus concluded that in contrast to Albucult, stripped rAlbumin better prevents the fibrillation of human insulin. Example 3a: ThT assay for Examples 4 to 9 and 14 to 25

ThT assays were performed using a BMG Fluostar Omega fluorescence plate reader equipped with excitation and emission filters of 440±10 nm and 490±10 nm, respectively. Cycles were repeated with a cycle time of 600 seconds with 300 seconds orbital shaking at a temperature of 50°C in 96 well microtiter plates (Nunc 265301 ) covered with a transparent film (Nunc 232701 ). The assay was stopped after 80 hours. An increase in fluorescence correlates with the formation of amyloid fibrils in the sample.

Example 3b: SE-HPLC assay

Quantification of glucagon was performed using a modified size exclusion high performance liquid chromatography (SE-HPLC) method described by Fang et al (Pharm Res (2012) 29:3278-3291 ). Dionex system equipped with P680 HPLC Pump, ASI-100 Automated Sample Injector, Thermostatted Column Compartment TCC-100 and PDA-100 Photodiode Array Detector was used. Separation of glucagon from albumin was achieved by using TSK gel G3000 SWXL (5 μπι (micron), 7.8 x 300 mm) size exclusion column. TSK gel SWXL Guard (7 μηη, 6.0 x 40 mm) column was used before the main column. Mobile phase consisted of 3.2 mM HCI and 100 mM NaCI, pH 2.5. Injection volume was 10 μΙ , flow rate was 1 ml/min and detection was done at 280 nm. Samples were placed in the sample compartment (25 °C) and were analyzed every 12 hours. In general, a lower AUC value reflects a lower amount of soluble glucagon.

Example 4; Affect of albumin on insulin stability

The affect of four different formulations of recombinant albumin on the stability of insulin and two insulin analogs were tested.

Insulin: A chain: SEQ ID NO: 3, B chain: SEQ ID NO: 4; Humalog (insulin lispro): A chain: SEQ ID NO: 25, B chain: SEQ ID NO: 26; Levemir (insulin detemir): A chain: SEQ ID NO: 30, B chain: SEQ ID NO: 31 ; 14-C fatty acid (myristic acid) is bound to the Lys at position 29 of the B chain. The 14-C fatty acid increases self-association and albumin binding. Recombumin® Prime (Novozymes Biopharma): 200 g/L recombinant human albumin (SEQ ID NO: 1 ), 145 mM sodium, 32 mM octanoate, 15 mg/L polysorbate 80, water for injection to 1 L.

Recombumin® Alpha (Novozymes Biopharma): 100 g/L recombinant human albumin (SEQ ID NO: 1 ), 145 mM sodium, 16 mM octanoate, 100 mg/L polysorbate 80, water for injection to 1 L.

Albix (Novozymes Biopharma): 100 g/L recombinant human albumin (SEQ ID NO: 1 ), 250 mM sodium, up to 0.2 mM octanoate, substantially free of polysorbate 80, water for injection to 1 L.

Stripped Recombumin® Alpha: Recombumin® Alpha (Albucult) stripped as described in Example 1.

Briefly, the ThT assay was carried out as described in Example 3a. The results are shown in Table 1 where a longer lag-time indicates a higher stability. '-' means that the sample did not fibrillate.

Table 1 : Lag time (hours) for three different insulins when stabilized with four different formulations of albumin.

The data of Table 1 show that stability of insulin, or analog, is increased by the presence of albumin with a relatively low level of fatty acid (e.g. octanoate) and/or a relatively low level of detergent (e.g. polysorbate 80).

Example 5: Affect of albumin concentration on stability of insulin and insulin analogs

Each sample contained 0.33 mg/ml (0.057 mM) insulin in 50 mM NaCI and 50 mM phosphate, pH 7.0 with a total sample volume of 200 μί. Five different albumin (Albix) concentrations (3.7 mg/ml - 57μΜ, 1 .3 mg/ml - 19μΜ, 0.76 mg/ml - 1 1 .4μΜ, 1 1.1 mg/ml - 171 μΜ and 18.5 mg/ml - 285 μΜ) were tested corresponding to a 1 :1 , 3:1 , 5:1 , 1 :3 and 1 :5 molar ratio of insulin to albumin. Two different types of insulin were tested (human insulin (A chain: SEQ ID NO: 3, B chain: SEQ ID NO: 4) and insulin lispro (A chain: SEQ ID NO: 25, B chain: SEQ ID NO: 26)) using the ThT assay of Example 3a. Prior to incubation in the plate reader, 20 μΙ_ 10 mM ThT solution was added yielding a total volume of 220 μΙ_ and a final ThT concentration of 1 mM. Each sample was tested in duplicate, both results are shown. Table 2 presents the presence (+) and absence (-) of fibrils after 80 hours and Table 3 shows the lag time in hours. The threshold used for fibril formation was, in both cases, 5000 relative fluorescence units (RFU). In Table 3 '-' means that the RFU did not exceed 5000 RFU during the experiment. Figure 3 shows the raw data which were used for calculation of the lag times in Table 3.

Table 2: Fibrillation of insulin and insulin lispro after 80 hours

The data show that the presence of albumin increases the stability of human insulin and insulin lispro.

Example 6: Affect of albumin formulation (octanoate and polysorbate 80 concentration) on stability of insulin

The affect of octanoate and polysorbate 80 concentration on the stability of insulin was studied using the ThT assay of Example 3a.

Each sample contained 0.55 mg/ml (0.095 mM) human insulin in 47.5 mM NaCI and

47.5 mM phosphate, pH 7.0 with a total sample volume of 200 μΙ_. The albumin (Albix) concentration was 1.9 mg/ml (0.029 mM) resulting in a 3.3:1 molar ratio of insulin to albumin in the assay. Three different octanoate concentrations (152 μΜ, 304 μΜ and 456 μΜ) were tested together with six different polysorbate 80 concentrations (0.095 mg/L, 0.19 mg/L, 0.285 mg/L, 0.475 mg/L, 0.570 mg/L and 0.665 mg/L). The octanoate concentrations result in a 5:1 , 1 1 :1 and 16:1 molar ratio of octanoate to albumin. Prior to incubation in the plate reader, 20 μί 10 mM ThT solution was added yielding a total volume of 220 μί and a final ThT concentration of 1 mM. Each sample was tested in duplicate, both results are shown. Table 4 presents the presence of fibrils after 80 hours and Table 5 shows the lag time in hours. The threshold used for fibril formation was in both cases 5000 RFU. In Table 5 '-' means that the RFU did not exceed 5000 RFU during the experiment, 'n/t' means not tested.

Table 4: Fibrillation of insulin after 80 hours

Table 5: Lag time (h) for insulin to reach threshold of 5000 RFU

Human insulin alone (33 / 35)

The data show that decreasing the amount of fatty acid (e.g. octanoate) decreases fibrillation of insulin stabilized with 1.9 mg/ml albumin and that decreasing the amount of detergent (e.g. polysorbate 80) decreases fibrillation of insulin stabilized with 1.9 mg/ml albumin when fatty acid (e.g. octanoate) is present in the formulation Example 7: Affect of albumin formulation (octanoate and polysorbate 80 concentration) on stability of insulin

Each sample contained 0.55 mg/ml (0.095 mM) human insulin in 47.5 mM NaCI and

47.5 mM phosphate, pH 7.0 with a total sample volume of 200 μΙ_. The albumin (Albix) concentration was 9.5 mg/ml (0.143 mM) resulting in a 1 : 1 .5 molar ratio of insulin to albumin in the assay. Three different octanoate concentrations (0.760 mM, 1 .52 mM and 2.28 mM) were tested together with six different polysorbate 80 concentrations (0.475 mg/L, 0.95 mg/L, 1 .425 mg/L, 2.375 mg/L, 2.85 mg/L and 3.325 mg/L). The octanoate concentrations result in a 5: 1 , 1 1 : 1 and 1 6: 1 molar ratio of octanoate to albumin . Prior to incubation in the plate reader, 20 [iL 10 mM ThT solution was added yielding a total volume of 220 L and a final ThT concentration of 1 mM. Each sample was tested in duplicate, both results are shown. Table 6 presents the presence (+) or absence (-) of fibrils after 80 hours and Table 7 shows the lag time in hours. The threshold used for fibril formation was in both cases 5000 RFU. 'n/t' means not tested. In Table 7 '-' means that the RFU did not exceed 5000 RFU during the experiment.

Table 6: Fibrillation of insulin after 80 hours

Table 7: Lag time (h) for insulin to reach threshold of 5000 RFU

Human insulin alone (33 / 35)

The data show that decreasing the amount of fatty acid (e.g. octanoate) decreases fibrillation of insulin stabilized with 9.5 mg/ml albumin and that decreasing the amount of detergent (e.g. polysorbate 80) decreases fibrillation of insulin stabilized with 9.5 mg/ml albumin when fatty acid (e.g. octanoate) is present in the formulation

Example 8: Affect of albumin formulation (octanoate and polysorbate 80 concentration) on stability of insulin lispro

The affect of octanoate and polysorbate 80 concentration on the stability of insulin lispro was studied using the ThT assay of Example 3a.

Each sample contained 0.73 mg/ml (0.125 mM) insulin lispro in 47.5 mM NaCI and 47.5 mM phosphate, pH 7.0 with a total sample volume of 200 μΙ_. The albumin (Albix) concentration was 1 .9 mg/ml (0.029 mM) resulting in a 4.3:1 molar ratio of insulin to albumin in the assay. Three different octanoate concentrations (152 μΜ, 304 μΜ and 456 μΜ) were tested together with six different polysorbate 80 concentrations (0.095 mg/L, 0.19 mg/L, 0.285 mg/L, 0.475 mg/L, 0.570 mg/L and 0.665 mg/L). The octanoate concentrations result in a 5:1 , 1 1 :1 and 16:1 molar ratio of octanoate to albumin. Prior to incubation in the plate reader, 20 μί 10 mM ThT solution was added yielding a total volume of 220 μί and a final ThT concentration of 1 mM. Each sample was tested in duplicate, both results are shown. Table 8 presents the presence (+) and absence (-) of fibrils after 80 hours and Table 9 shows the lag time in hours. The threshold used for fibril formation was in both cases 5000 relative fluorescence units (RFU). 'n/t' means not tested. In Table 9 '-' means that the RFU did not exceed 5000 RFU during the experiment. Table 8: Fibrillation of insulin after 80 hours

Table 9: Lag time (h) for insulin to reach threshold of 5000 RFU

Insulin lispro alone (37 / 39)

The data show that decreasing the amount of fatty acid (e.g. octanoate) decreases fibrillation of insulin stabilized with 1.9 mg/ml albumin and that decreasing the amount of detergent (e.g. polysorbate 80) decreases fibrillation of insulin stabilized with 1 .9 mg/ml albumin when fatty acid (e.g. octanoate) is present in the formulation

Example 9: Affect of albumin formulation (octanoate and polysorbate 80 concentration) on stability of insulin lispro

Each sample contained 0.73 mg/ml (0.125 mM) insulin lispro in 47.5 mM NaCI and 47.5 mM phosphate, pH 7.0 with a total sample volume of 200 μί. The albumin (Albix) concentration was 9.5 mg/ml (0.143 mM) resulting in a 1 :1.15 molar ratio of insulin to albumin in the assay. Three different octanoate concentrations (0.760 mM, 1 .52 mM and 2.28 mM) were tested together with six different polysorbate 80 concentrations (0.475 mg/L, 0.95 mg/L, 1.425 mg/L, 2.375 mg/L, 2.85 mg/L and 3.325 mg/L). The octanoate concentrations result in a 5:1 , 1 1 :1 and 16:1 molar ratio of octanoate to albumin. Prior to incubation in the plate reader, 20 μΙ_ 10 mM ThT solution was added yielding a total volume of 220 μΙ_ and a final ThT concentration of 1 mM. Each sample was tested in duplicate, both results are shown. Table 10 presents the presence (+) and absence (-) of fibrils after 80 hours and Table 1 1 shows the lag time in hours.

The threshold used for fibril formation was in both cases 5000 RFU. 'n/t' means not tested. In Table 1 1 '-' means that the RFU did not exceed 5000 RFU during the experiment.

Table 10: Fibrillation of insulin after 80 hours

The data show that decreasing the amount of fatty acid (e.g. octanoate) decreases fibrillation of insulin stabilized with 9.5 mg/ml albumin and that decreasing the amount of detergent (e.g. polysorbate 80) decreases fibrillation of insulin stabilized with 9.5 mg/ml albumin when fatty acid (e.g. octanoate) is present in the formulation.

Example 10: Affect of albumin concentration on stability of glucagon

The affect of albumin concentration on the stability of glucagon was tested in 100 mM phosphate buffer (pH5) at 25 °C by SE-HPLC as described in Example 3b. Each sample (250 μΙ) contained 1 mg/ml (287 uM) glucagon. Four different albumin (Albix) concentrations (1 mg/ml (15 uM), 5 mg/ml, 10 mg/ml, 20 mg/ml) were tested. Figure 4 presents the area under the curve plotted over the period of time. All results are presented as an average (mean) of three, error bars are standard deviation.

Figure 4 shows that the stability of glucagon is increased by the presence of increasing concentrations of albumin.

Example 11 : Affect of fatty acid and detergent on the stability of glucagon stabilized by albumin

The affect of fatty acid (octanoate) and detergent (polysorbate 20 and polysorbate 80) on the ability of albumin to stabilize glucagon was studied in 100 mM phosphate buffer (pH5) at 25 °C by SE-HPLC as described in Example 3b.

Each sample (250 μΙ) contained 1 mg/ml glucagon. Polysorbate 80 (0.05%), polysorbate 20 (0.05%), octanoate (0.8 mM) and 20 mg/ml Albix were tested individually. Figure 5 presents the area under the curve plotted over the period of time. All results are presented as an average (mean) of three measurements, error bars are standard deviation.

Figure 6 shows that the addition of 0.8 mM octanoate or 0.05% detergent adversely affects the stabilizing effect of albumin on glucagon. Albix contains low levels of octanoate and substantially no detergent and, when added to glucagon, showed a good stabilizing effect.

Example 12: Affect of buffer on stability of glucagon stabilized by albumin

The affect of buffer type on the ability of albumin to stabilize glucagon (SEQ ID NO: 8) was studied at pH 5, 25 °C by SE-HPLC using the method of Example 3b.

Each sample (250 μΙ) contained 1 mg/ml glucagon and 20 mg/ml albumin. Three different buffers were used: 100 mM citrate buffer pH 5 (prepared using 1 M solution of sodium citrate dihydrate and titrating with 1 M citric acid to pH 5 and diluting it 10-fold to obtain final buffer strength); 100 mM phosphate buffer pH 5 (prepared using 1 M solution of NaH₂P0₄.H₂0 and titrating with 1 M Na₂HP0₄.2H₂0 to pH 5 and diluting it 10-fold to obtain final buffer strength) and 40 mM histidine buffer pH 5 (prepared using 400 mM solution of L-histidine, adjusting final pH to 5 and diluting it 10-fold to obtain final buffer strength).

Figure 7 presents the area under the curve plotted over the period of time. All results are presented as an average (mean) of three measurements, error bars are standard deviation.

The data show that while all of phosphate, citrate and histidine buffers stabilize glucagon in combination with albumin, phosphate and histidine are preferred.

Example 13: Affect of amino acids on the ability of albumin to stabilize glucagon

The affect of amino acids on the ability of albumin to stabilize glucagon (SEQ ID NO: 8) was studied at pH 5, 25 °C in 100 mM phosphate buffer by SE-HPLC using the method of Example 3b. Each sample (250 μΙ) contained 1 mg/ml glucagon. Three amino acids (at a concentration of 20 mg/ml (methionine (134 mM), glycine (266 mM) and arginine (1 15 mM)) were tested either alone or along with 20 mg/ml Albix. Table 12 summarizes the data obtained after 72 h. Results are presented as an average of three measurements for samples with albumin while average of two measurements for samples (control samples) without albumin (Mean ± standard deviation (SD)).

Table 12

The data of Table 12 show that the presence of arginine or glycine has the most significant ffect on the ability of albumin to stabilize glucagon.

Example 14: Affect of pH on stabililty of GLP-2 analog

The affect of pH on the ability of albumin to stabilize a GLP-2 analog (teduglutide, SEQ ID NO: 28) was studied using the ThT assay of Example 3a, with the exception that the temperature was 40 °C.

Each sample (200 μΙ) contained a final concentration of 0.9 mg/ml teduglutide. Five different phosphate buffers were prepared with a final concentration of 100 mM each (prepared by using 1 M solutions of NaH₂P0₄.H₂0 and titrating with 1 M Na₂HP0₄.2H₂0 to obtain pH 4, pH 5, pH 6, pH 7 and pH 8; each buffer was then diluted 10-fold to obtain final buffer strength). Albucult and Albix concentrations were 20 mg/ml in the final test conditions. ThT was used at a final concentration of 1 mM. The threshold used for fibril formation was 10000 RFU.

Table 13 shows the affect of pH 4, 5, 6, 7 and 8 on the ability of two different albumin formulations (Albucult and Albix) to stabilize teduglutide. Table 13: Lag time (h) of teduglutide (GLP-2 analog) to form fibrils (time to reach threshold of

10000 RFU in THT assay)

Generally, the data show that Albix provides a better stabilizing effect than Albucult. This suggests that the presence of increased levels of fatty acid (e.g. octanoate) and/or detergent (e.g. polysorbate 80) has a detrimental affect on the ability of albumin to stabilize the teduglutide. The best stability is achieved at pH 7 or above.

Example 15: Affect of albumin concentration on stability of GLP-2 analog

The affect of albumin concentration on stability of a GLP-2 analog (teduglutide, SEQ ID

NO: 28) was studied using the ThT assay of Example 3a, with the exception that the temperature was 40 °C.

Each sample (200 μΙ) contained a final concentration of 1.74 mg/ml teduglutide. Final test solution was buffered by using NaH₂P0₄.H₂0 (0.57 mg/ml), Na₂HP0₄.2H₂0 (2.02 mg/ml) and L-Histidine (3.45 mg/ml). Three different Albix concentrations (5 mg/ml, 10 mg/ml and 20 mg/ml) were tested. ThT was used at a final concentration of 1 mM. The threshold used for fibril formation was 10000 RFU.

Figure 9 shows that all of 5, 10, and 20 mg/ml albumin stabilized teduglutide. Example 16: Affect of amino acids on the ability of albumin to stabilize GLP-2 analog

The affect of amino acids on the ability of albumin to stabilize GLP-2 analog (teduglutide, SEQ ID NO: 28) was studied by using the ThT assay of Example 3a, with the exception that the temperature was 40 °C.

Each sample (200 μΙ) contained a final concentration of 1.74 mg/ml teduglutide. Final test solution was buffered by using NaH₂P0₄.H₂0 (0.57 mg/ml), Na₂HP0₄.2H₂0 (2.02 mg/ml) and L-Histidine (3.45 mg/ml). Four different amino acids (glycine, arginine, lysine and methionine) were tested at a final concentration of 20 mg/ml. Test concentration of Albix was 20 mg/ml. ThT was used at a final concentration of 1 mM. The threshold used for fibril formation was 10000 RFU.

The data of Fig. 10 show that the presence of albumin was sufficient to stabilize teduglutide. All the formulations containing albumin and amino acids were also able to stabilize the GLP-2 analog. Example 17: Affect of octanoate and polysorbate concentration on the ability of albumin to stabilize GLP-2 analog.

The affect of octanoate and polysorbate 80 concentration on the stability of a GLP-2 analog was studied using the ThT assay of Example 3a. Each sample contained 0.21 mg/ml (0.055 mM) Teduglutide (GLP-2 analog, SEQ ID NO: 28) in 25 mM NaCI and 25 mM phosphate, pH 7.0 with a total sample volume of 200 μί. The albumin (Albix) concentration was 1 mg/ml (0.015 mM) resulting in a 3.7:1 molar ratio of teduglutide to albumin in the assay. Three different octanoate concentrations (80 μΜ, 160 μΜ and 240 μΜ) were tested together with two different polysorbate 80 concentrations (0.05 mg/L and 0.10 mg/L). The octanoate concentrations result in a 5:1 , 11 :1 and 16:1 molar ratio of octanoate to albumin. Prior to incubation in the plate reader, 20 μί 10 mM ThT solution was added yielding a total volume of 220 μί and a final ThT concentration of 1 mM. Each sample was tested in duplicate, both results are shown. Table 14 presents the presence (+) or absence (-) of fibrils after 80 hours and Table 15 shows the lag time in hours. The threshold used for fibril formation was in both cases 5000 RFU. 'n/t' means not tested. In Table 15 '-' means that the RFU did not exceed 5000 RFU during the experiment.

Table 14: Fibrillation of teduglutide after 80 hours

Table 15: Lag time (h) for teduglutide to reach threshold of 5000 RFU

Teduglutide analog alone (1 / 1 )

The data show that a lower detergent, e.g. polysorbate 80, concentration is desirable stabilizing GLP-2 analogs such as teduglutide. The data show that a lower fatty acid, e.g. octanoate, concentration is desirable stabilizing GLP-2 analogs such as teduglutide Example 18: Affect of octanoate and polysorbate concentration on the ability of albumin to stabilize GLP-2 analog.

The affect of octanoate and polysorbate 80 concentration on the stability of a GLP-2 analog was studied using the ThT assay of Example 3a.

Each sample contained 0.21 mg/ml (0.055 mM) GLP-2 analog (Teduglutide, SEQ I D NO: 28) in 25 mM NaCI and 25 mM phosphate, pH 7.0 with a total sample volume of 200 μ ί. The albumin (Albix) concentration was 5 mg/ml (0.075 mM) resulting in a 1 : 1 .4 molar ratio of teduglutide to albumin in the assay. Three different octanoate concentrations (400 μΜ, 800 μ Μ and 1200 μΜ) were tested together with two different polysorbate 80 concentrations (0.25 mg/L and 0.50 mg/L). The octanoate concentrations result in a 5:1 , 1 1 : 1 and 16: 1 molar ratio of octanoate to albumin. Prior to incubation in the plate reader, 20 [iL 10 mM ThT solution was added yielding a total volume of 220 μί and a final ThT concentration of 1 mM. Each sample was tested in duplicate, both results are shown . Table 16 presents the presence (+) or absence (-) of fibrils after 80 hours and Table 17 shows the lag time in hours. The threshold used for fibril formation was in both cases 5000 RFU . 'nit' means not tested. I n Table 1 7 '-' means that the RFU did not exceed 5000 RFU during the experiment.

Table 1 6: Fibrillation of teduglutide after 80 hours

Table 1 7: Lag time (h) for teduglutide to reach threshold of 5000 RFU

Teduglutide alone (1 / 1 )

The data show that a lower fatty acid, e.g. octanoate, concentration is desirable for stabilizing GLP-2 analogs such as teduglutide. Example 19: Affect of fatty acid and detergent on the ability of albumin to stabilize GLP-1 analog

The affect of octanoate and polysorbate 80 concentration on the stability of a GLP-1 analog was studied using the ThT assay of Example 3a.

Each sample contained 0.26 mg/ml (0.077 mM) liraglutide (GLP-1 analog, SEQ I D NO:

27) in 25 mM NaCI and 25 mM phosphate, pH 7.0 with a total sample volume of 200 μ ί. The albumin (Albix) concentration was 1 mg/ml (0.015 mM) resulting in a 5.2: 1 molar ratio of liraglutide to albumin in the assay. Three different octanoate concentrations (80 μΜ, 1 60 μΜ and 240 μΜ) were tested together with two different polysorbate 80 concentrations (0.05 mg/L and 0.10 mg/L). The octanoate concentrations result in a 5:1 , 1 1 : 1 and 16: 1 molar ratio of octanoate to albumin. Prior to incubation in the plate reader, 20 [iL 10 mM ThT solution was added yielding a total volume of 220 L and a final ThT concentration of 1 mM. Each sample was tested in duplicate, both results are shown . Table 18 presents the presence (+) or absence (-) of fibrils after 80 hours and Table 19 shows the lag time in hours. The threshold used for fibril formation was in both cases 5000 RFU . 'n/t' means not tested. I n Table 1 9 '-' means that the RFU did not exceed 5000 RFU during the experiment.

Table 1 8: Fibrillation of liraglutide after 80 hours

Table 19: Lag time (h) for liraglutide to reach threshold of 5000 RFU

Liraglutide alone (50 / 49)

The data show that a lower fatty acid, e.g. octanoate, concentration is desirable for stabilizing GLP-1 analogs such as liraglutide

Example 20: Affect of fatty acid and detergent on the ability of albumin to stabilize GLP-1 analog

The affect of octanoate and polysorbate 80 concentration on the stability of a GLP-1 analog was studied using the ThT assay of Example 3a. Each sample contained 0.26 mg/ml (0.077 mM) liraglutide (GLP-1 analog, SEQ ID NO: 27) in 25 mM NaCI and 25 mM phosphate, pH 7.0 with a total sample volume of 200 μΙ_. The albumin (Albix) concentration was 5 mg/ml (0.075 mM) resulting in a 1 :1 molar ratio of liraglutide to albumin in the assay. Three different octanoate concentrations (400 μΜ, 800 μΜ and 1200 μΜ) were tested together with two different polysorbate 80 concentrations (0.25 mg/L and 0.50 mg/L). The octanoate concentrations result in a 5:1 , 1 1 :1 and 16:1 molar ratio of octanoate to albumin. Prior to incubation in the plate reader, 20 μΙ_ 10 mM ThT solution was added yielding a total volume of 220 μΙ_ and a final ThT concentration of 1 mM. Each sample was tested in duplicate, both results are shown. Table 20 presents the presence (+) or absence (-) of fibrils after 80 hours and Table 21 shows the lag time in hours. The threshold used for fibril formation was in both cases 5000 RFU. 'n/t' means not tested. In Table 21 '-' means that the RFU did not exceed 5000 RFU during the experiment.

Table 20: Fibrillation of liraglutide after 80 hours

Table 21 : Lag time (h) for liraglutide to reach threshold of 5000 RFU

Liraglutide alone (50 / 49)

The data show that a lower lower fatty acid, e.g. octanoate 80, concentration is desirable for stabilizing GLP-1 analogs such as liraglutide. The data show that a lower detergent, e.g.

polysorbate 80, concentration is desirable for stabilizing GLP-1 analogs such as liraglutide.

Example 21 : Affect of pH on the ability of albumin to stabilize HIV fusion inhibitor

The affect of pH on the ability of albumin to stabilize enfuvirtide (SEQ ID NO: 29) was studied using the ThT assay of Example 3a, with the exception that the temperature was 40 °C.

Each sample (200 μΙ) contained a final concentration of 42.7 mg/ml enfuvirtide. Five different phosphate buffers were prepared with a final concentration of 100 mM each (prepared using 1 M solution of NaH₂P0₄.H₂0, titrating with 1 M Na₂HP0₄.2H₂0 to obtain pH 4, pH 5, pH

6, pH 7 and pH 8; each buffer was then diluted 10-fold to obtain final buffer strength). Albucult and Albix concentrations were 20 mg/ml in the final test conditions. ThT was used at a final concentration of 1 mM. The threshold used for fibril formation was 20000 RFU.

Table 22 shows the affect of pH 4, 5, 6, 7 and 8 on the ability of two different albumin formulations (Albucult and Albix) to stabilize enfuvirtide.

Table 22: Lag time (h) for HIV fusion inhibitor to form fibrils (time to reach threshold of 20000 RFU in THT assay)

Generally, Albix provides a better stabilizing effect than Albucult. A pH of 6 or above is beneficial and that pH 8 is particularly beneficial. Enfuvirtide is normally formulated at pH 9. The data show that albumin allows stabilization at lower pH. Formulating at a lower pH is desirable because it improves patient comfort.

Example 22: Affect of albumin concentration on stability of HIV fusion inhibitor.

The affect of albumin concentration on the stability of enfuvirtide ("Fuzeon", SEQ ID NO:

29) was studied using the ThT assay of Example 3a, with the exception that the temperature was 40 °C.

Each sample (200 μΙ) contained a final concentration of 39.9 mg/ml enfurvitide in phosphate buffer. Phosphate buffer (100 mM, pH 8) was prepared by using 1 M solution of Na₂HP0₄.2H₂0, titrating with 1 M NaH₂P0₄.H₂0 to obtain pH 8; which was then diluted 10-fold to obtain final buffer strength. Three different Albix concentrations (5 mg/ml, 10 mg/ml and 20 mg/ml) were tested. ThT was used at a final concentration of 1 mM. The threshold used for fibril formation was 20000 RFU.

Figure 1 1 shows that the stability of enfuvirtide is increased by increasing concentrations of albumin.

Example 23: Affect of amino acids on the ability of albumin to stabilize HIV fusion inhibitor

The affect of amino acids on the ability of albumin to stabilize enfuvirtide ("Fuzeon", SEQ ID NO: 29) was studied using the ThT assay of Example 3a, with the exception that the temperature was 40 °C.

Each sample (200 μΙ) contained a final concentration of 39.9 mg/ml enfurvitide. Phosphate buffer (100 mM, pH 8) was used, and prepared according to Example 22. Four different amino acids (glycine, arginine, lysine and methionine) were tested at a final concentration of 20 mg/ml. Albumin (Albix) concentration was 20 mg/ml in the test conditions. ThT was used at a final concentration of 1 mM. The threshold used for fibril formation was 20000 RFU.

The data of Table 23 show that addition of glycine improves the ability of albumin to stabilize enfuvirtide.

Table 23: Lag time (h) for enfuvirtide to form fibrils (time to reach threshold of 20000 RFU in

THT assay)

Example 24: Affect of octanoate and polysorbate 80 on the ability of albumin to stabilize HIV fusion inhibitor

The affect of octanoate and polysorbate 80 concentration on the stability of an HIV fusion inhibitor was studied using the ThT assay of Example 3a.

Each sample contained 4.8 mg/ml (1 .08 mM) enfuvirtide SEQ ID NO: 29) in 45 mM NaCI and 45 mM phosphate, pH 7.0 with a total sample volume of 200 μί. The albumin (Albix) concentration was 1.8 mg/ml (0.027 mM) resulting in a 40:1 molar ratio of enfuvirtide to albumin in the assay. Three different octanoate concentrations (144 μΜ, 288 μΜ and 432 μΜ) were tested together with two different polysorbate 80 concentrations ( 0.09 mg/L and 0.18 mg/L). The octanoate concentrations result in a 5:1 , 11 :1 and 16:1 molar ratio of octanoate to albumin. Prior to incubation in the plate reader, 20 μί 10 mM ThT solution was added yielding a total volume of 220 μί and a final ThT concentration of 1 mM. Each sample was tested in duplicate, both results are shown. Table 24 shows the lag time in hours, 'n/t' means not tested. The threshold used for fibril formation was in both cases 10000 RFU. Table 24: Lag time (h) for enfuvirtide to reach threshold of 10000 RFU

The data show that while albumin is useful to stabilize enfuvirtide, increasing the amount of fatty acid (e.g. octanoate) hinders the stabilizing effect.

Example 25 Affect of octanoate and polysorbate on the ability of albumin to stabilize HIV fusion inhibitor

Each sample contained 4.8 mg/ml (1 .08 mM) enfuvirtide (SEQ ID NO: 29) in 45 mM NaCI and 45 mM phosphate, pH 7.0 with a total sample volume of 200 μί. The albumin (Albix) concentration was 9 mg/ml (0.136 mM) resulting in a 8: 1 molar ratio of enfuvirtide to albumin in the assay. Three different octanoate concentrations (720 μΜ, 1440 μΜ and 2160 μΜ) were tested together with two different polysorbate 80 concentrations (0.45 mg/L and 0.90 mg/L). The octanoate concentrations result in a 5: 1 , 1 1 : 1 and 16:1 molar ratio of octanoate to albumin. Prior to incubation in the plate reader, 20 μί 10 mM ThT solution was added yielding a total volume of 220 μί and a final ThT concentration of 1 mM. Each sample was tested in duplicate, both results are shown. Table 25 shows the lag time in hours, 'n/t' means not tested. The threshold used for fibril formation was in both cases 10000 RFU.

Table 25: Lag time (h) for enfuvirtide to reach threshold of 10000 RFU

Overview of sequences

SEQ ID NO. 1 : Native HSA SEQ ID NO. 2: Immature human insulin

SEQ ID NO. 3: Human insulin A chain

SEQ ID NO. 4: Human insulin B chain

SEQ ID NO. 5: Human glucagon like peptide 1 , Corresponding to amino acids 7-36 of Human GLP1 ; 3.3 kDa, 31 amino acids

SEQ ID NO. 6: Human glucagon like peptide 2

SEQ ID NO. 7: Human growth hormone

SEQ ID NO. 8: Human Glucagon

SEQ ID NO. 9: cDNA encoding HSA

SEQ ID NO. 10: Albumin - Pan troglodytes

SEQ ID NO. 11 : Albumin - Macaca mulatta

SEQ ID NO. 12: Albumin - Mesocricetus auratus

SEQ ID NO. 13: Albumin - Cavia porcellus

SEQ ID NO. 14: Albumin - Mus musculus

SEQ ID NO. 15: Albumin - Rattus norvegicus

SEQ ID NO. 16: Albumin - Bos taurus

SEQ ID NO. 17: Albumin - Equus caballus

SEQ ID NO. 18: Albumin - Equus asinus

SEQ ID NO. 19: Albumin - Oryctolagus cuniculus

SEQ ID NO. 20: Albumin - Capra hircus

SEQ ID NO. 21 : Albumin - Ovis aries

SEQ ID NO. 22: Albumin - Canis lupus familiaris

SEQ ID NO. 23: Albumin - Gallus gallus

SEQ ID NO. 24: Albumin - Sus scrofa

SEQ ID NO. 25: Lispro insulin analog A chain

SEQ ID NO. 26: Lispro insulin analog B chain

SEQ ID NO. 27: Liraglutide GLP-1 analog

SEQ ID NO. 28: Teduglutide GLP-2 analog

SEQ ID NO. 29: Enfuvirtide HIV fusion inhibitor

Claims

A composition comprising:

a. from 0.01 mg/ml to 300 mg/ml peptide, the peptide having a size of from 10 to 100 amino acids;

b. from 0.01 mg/ml to 500 mg/ml albumin

A composition comprising:

b. from 0.01 mg/ml to 500 mg/ml albumin

The composition according to claim 1 comprising less than or equal to 25 mM octanoate and less than or equal to 0.001 % (w/v) polysorbate 80.

The composition according to any preceding claim, wherein the peptide is a non- lipopeptide.

The composition according to any preceding claim, wherein the peptide is essentially in non-fibril form.

The composition according to any preceding claim, wherein the peptide comprises two or more different peptide chains and/or the composition comprises two or more different peptides.

The composition according to any preceding claim, wherein the peptide is selected from the group consisting of insulin, insulin analogs, glucagon, GLP-1 or analog thereof, GLP- 2 or analog thereof, HIV fusion inhibitors or a fragment or variant thereof.

The composition according to any preceding claim, wherein the peptide comprises less than 95 amino acid residues, such as less than 90 amino acid residues, such as less than 85 amino acid residues, such as less than 80 amino acid residues, such as less than 75 amino acid residues, such as less than 70 amino acid residues, such as less than 65 amino acid residues, such as less than 60 amino acid residues, such as less than 55 amino acid residues, such as less than 50 amino acid residues, such as less than 45 amino acid residues, such as less than 40 amino acid residues, such as less than 35 amino acid residues, such as less than 30 amino acid residues, such as less than 25 amino acid residues, such as less than 20 amino acid residues, such as less than 15 amino acid residues.

The composition according to any of claims 2 to 8 wherein the less than 0.001 % (w/v) detergent is less than 0.001 % (w/v) non-ionic detergent and/or less than 0.001 % (w/v) anionic detergent and/or less than 0.001 % (w/v) cationic detergent and/or less than 0.001 % (w/v) zwitterionic detergent.

10. The composition according to claim 9, wherein the less than 0.001 % (w/v) non-ionic detergent is less than 0.001 % (w/v) polysorbate 80 and/or less than 0.001 % (w/v) polysorbate 20 and/or less than 0.001 % (w/v) poloxamer.

1 1. The composition according to any preceding claim, wherein the composition is essentially detergent free.

12. The composition according to any preceding claim, wherein the less than or equal to 25 mM fatty acids is less than or equal to 25 mM octanoate.

13. The composition according to claim 12, wherein the less than or equal to 25 mM octanoate or total fatty acids is less than or equal to 20 mM octanoate or total fatty acids, such as less than or equal to 15 mM octanoate or total fatty acids, such as less than or equal to 10 mM octanoate or total fatty acids, such as less than or equal to 5 mM octanoate or total fatty acids, such as less than or equal to 2 mM octanoate or total fatty acids, such as less than or equal to 1 mM octanoate or total fatty acids.

14. The composition according to any preceding claim, wherein the composition is essentially octanoate free or substantially free of total fatty acids.

15. The composition according to any preceding claim, wherein the composition comprises free glycine or free arginine.

16. The composition according to claim 15 wherein the concentration of free glycine or free or free arginine is at least 5 mg/ml, such as at least 10 mg/ml, such as at least 20 mg/ml. 17. The composition according to any preceding claim, wherein the composition comprises less than or equal to 25 mM amphiphilic compounds.

18. The composition according to any preceding claim, wherein the composition is essentially free from amphiphilic compounds.

19. The composition according to any preceding claim, wherein the molar ratio of peptide to albumin is from 1 part peptide to 2000 parts albumin (1 : 2000) to 3000 parts peptide to 1 part albumin (3000: 1 ).

20. The composition according to any preceding claim, wherein the molar ratio of octanoate to albumin is less than or equal to 16:1.

21. The composition according to any preceding claim, wherein the composition is essentially free of zinc.

22. A composition which is essentially free of zinc comprising:

a. from 0.01 mg/ml to 300 mg/ml insulin or insulin analog;

b. from 0.01 mg/ml to 500 mg/ml albumin

23. The composition according to any preceding claim, wherein the composition is essentially metal ion free.

24. The composition according to any preceding claim wherein said composition has a pH of between 4 and 9; such as between 4 and 8; such as between 4 and 7; such as between 5 and 8; such as between 6 and 8; preferably between 6.5 and 7.5 such as wherein said composition has a pH of about 7.

25. The composition according to claim 24 wherein the peptide is selected from insulin or an insulin analog and the pH is from about 4 to about 7 (preferably about 7).

26. The composition according to claim 25 wherein the molar ratio of insulin to albumin is from about 10:1 to about 1 :10, such as from about 5:1 to about 1 :5, such as about 3.3:1.

27. The composition according to claim 25 or 26 wherein the albumin concentration is from about 0.5 to about 20 mg/ml (such as from about 1 to about 19, such as from about 3.7 to about 18.5 mg/ml, such as from about 1.5 to about 12, such as from about 1 .7 to about 10) and the insulin concentration is from about 0.01 to about 1 mg/ml (such as from about 0.1 to about 1 , such as from such as from about 0.3 to about 0.8 mg/ml, such as from about 0.33 to about 0.73).

28. The composition according to claim 25, 26 or 27 wherein the molar ratio of octanoate to albumin is from about 1 :1 to about 20:1 such as from about 5:1 to about 16:1.

29. The composition according to any of claims 26 to 29 wherein the insulin analog is lispro.

30. The composition according to claim 24 wherein the peptide is selected from GLP-2 or GLP-2 analog and the pH is at least about 6, such as at least about 7, such as at least about 8, such as from about 6 to about 8.

31. The composition according to claim 30 wherein the molar ratio of GLP-2 or GLP-2 analog to albumin is from about 10:1 to about 1 :10, such as from about 5: 1 to about 1 :5, such as from about 5:1 to about 1 :2, such as from about 5: 1 to about 1 :1.4.

32. The composition according to claim 30 or 31 wherein the albumin concentration is from about 0.5 to about 20 mg/ml (such as from about 1 to about 10, such as from about 1 to about 5 mg/mlsuch as from about 0.1 to about 2 mg/ml, such as from about 0.2 to about 1 .8 mg/ml, such as from about 0.21 to about 1.74 mg/ml) and the GLP-2 or GLP-2 analog concentration is from about 0.1 to about 5 mg/ml (such as from about 0.21 to about 2.4 mg/ml).

33. The composition according to any of claims 30, 31 , or 32 wherein the molar ratio of octanoate to albumin is from about 1 :1 to about 20:1 such as from about 5:1 to about 16:1 .

34. The composition according to any of claims 30 to 33 wherein the GLP-2 analog is teduglutide.

35. The composition according to claim 34 wherein the peptide is a GLP-1 analog.

36. The composition according to claim 35 wherein the molar ratio of GLP-1 analog to albumin is from about 10:1 to about 1 :10, such as from about 7:1 to about 1 :7, such as from about 4:1 to about 6:1 , such as about 5.2:1.

37. The composition according to claim 35 or 36 wherein the albumin concentration is from about 0.5 to about 20 mg/ml (such as from about 1 to about 10, such as from about 1 to about 5 mg/ml) and the GLP-1 analog concentration is from about 0.2 to about 0.3 mg/ml (such as about 0.26 mg/ml).

38. The composition according to claim 35, 36, or 37 wherein the molar ratio of octanoate to albumin is from about 1 :1 to about 20:1 such as from about 5:1 to about 16:1.

39. The composition according to any of claims 35 to 38 wherein the GLP-1 analog is liraglutide.

40. The composition according to claim 24 wherein the peptide is an HIV inhibitor such as enfuvirtide or analog thereof and the pH is at least about 6, such as at least about 7, such as at least about 8, such as from about 6 to about 8.

41. The composition according to claim 40 wherein the wherein the molar ratio of enfuvirtide or analog thereof to albumin is from about 50:1 to about 1 :50, such as from about 45: 1 to about 7:1 , such as from about 39:1 to about 41 :1 or about 6:1 to about 8:1 , such as about 40:1 or about 8:1.

42. The composition according to claim 40 or 41 wherein the albumin concentration is from about 0.5 to about 20 mg/ml (such as from about 1 to about 10, such as from about 1 to about 5 mg/ml) and the enfuvirtide or analog thereof concentration is from about 1 to about 50 mg/ml (such as from about 4 to about 45 mg/ml, such as from about 48 to about 45 mg/ml, such as from about 38 to about 45 mg/ml, such as from about 39.9 to about 42.7 mg/ml).

43. The composition according to claim 41 , 42, or 42 wherein the molar ratio of octanoate to albumin is from about 1 :1 to about 20:1 such as from about 5:1 to about 16:1.

44. The composition according to claim 24 wherein the peptide is glucagon and the pH is from about 4.5 to about 5.5, preferably about 4.5 to about 5.5, most preferably about 5.0.

45. The composition according to claim 44, comprising arginine or glycine.

46. The composition according to any preceding claim wherein said composition consists essentially of albumin and insulin.

47. Use of highly purified albumin composition for preventing and/or reducing formation of insulin fibrils in an essentially zinc ion free formulation.

48. Use of the composition according to any one of claims 1 to 46 for preventing and/or reducing formation of peptide fibrils.

49. A method of stabilizing an essentially zinc ion free insulin composition comprising dissolving zinc ion free insulin in an aqueous solution of highly purified albumin, wherein the aqueous solution is essentially detergent free.

50. A method of preventing and/or reducing formation of peptide fibrils in an aqueous solution, the method comprising dissolving the peptide in an aqueous solution of albumin, wherein the aqueous solution is essentially free of amphiphilic compounds.

51. A method of preventing and/or reducing formation of peptide fibrils, the method comprising preparing a composition as defined in any of claims 1 to 44.