Therapeutic compositions and related methods

REFERENCE TO SEQUENCE LISTING

The present application is being filed along with a Sequence Listing in electronic format. The Sequence Listing is provided as a file entitled 2013-08-28 ST-001.02 ST_—25.txt, created Aug. 29, 2013, which is 176 KB in size. The information in the electronic format of the Sequence Listing is incorporated herein by reference in its entirety. The content of the sequence listing information recorded in computer readable form is identical to the written sequence listing.

FIELD OF THE INVENTION

The present invention generally relates to therapeutic compositions for the treatment of mammalian diseases and related methods. It more specifically relates to compositions comprising peptides for the treatment of inflammation and methods for delivering the peptides.

BACKGROUND OF THE INVENTION

Accumulation of amyloid fibrils correlates with many disease states. Such disease states include: Alzheimer's disease; diabetes mellitus type 2; Parkinson's disease; transmissible spongiform encephalopathy; Huntington's disease; medullary carcinoma of the thyroid; cardiac arrhythmias; atherosclerosis; rheumatoid arthritis; aortic medical amyloid; prolactinomas; familial amyloid polyneuropathy; hereditary non-neuropathic systemic amyloidosis; dialysis related amyloidosis; Finnish amyloidosis; lattice corneal dystrophy; cerebral amyloid angiopathy; systemic AL amyloidosis; and, sporadic inclusion body myositis.

The reason for a correlation between amyloid fibril accumulation and the specified diseases is unclear. Amyloid deposits physically disrupt tissue architecture in certain cases, which may impair function by a bulk process. Fibril deposition is associated with mitochondrial dysfunction, and the resulting initiation of apoptosis, in other cases. There have been some reports indicating that amyloid polymers can induce polymerization of essential amyloidogenic proteins, which is detrimental to cells.

SUMMARY OF THE INVENTION

In a composition aspect, the present invention provides a composition for treatment of one or more inflammatory conditions in a mammal. The composition comprises a peptide comprising amino acids of L-configuration, D-configuration or a mixture of configurations, wherein the peptide comprises one or more hexapeptides of a structure that generates a free energy of formation of −23 kcal/mol or less (i.e., more negative) when the hexapeptide sequence in subjected to the Rosetta-Profile algorithm. Example structures are provided below.

In another composition aspect, the present invention provides a composition for treatment of one or more inflammatory conditions in a mammal. The composition comprises a fibril. The fibril comprises four or more hexapeptides comprising amino acids of an L-configuration, D-configuration or a mixture of configurations having free energy of formation of −23 kcal/mol or less (i.e., more negative) when the hexapeptide sequence in subjected to the Rosetta-Profile algorithm.

In another composition aspect, the present invention provides a composition for treatment of one or more inflammatory conditions in a mammal. The composition comprises a fibril and a plasma protein. The fibril comprises four or more hexapeptides comprising amino acids of an L-configuration, D-configuration or a mixture of configurations having free energy of formation of −23 kcal/mol or less (i.e., more negative) when the hexapeptide sequence in subjected to the Rosetta-Profile algorithm.

In a method aspect, the present invention provides a method of treating inflammation comprising the step of administering one of the compositions discussed above.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 shows graphs related to the therapeutic effectiveness of amyloidogenic hexamers in a mouse model.

FIG. 2 shows bar graphs related to the modulation of plasma cytokine levels by certain amyloidogenic hexamers.

FIG. 3 shows bar graphs related to the pH dependency of amyloid formulation for certain amyloidogenic hexamers.

FIG. 4 shows graphs related to the correlation of ThT staining of amyloid fibrils with molecular chaperone function.

FIG. 5 shows bar graphs related to the binding of plasma proteins by amyloid fibrils.

FIG. 6 shows various graphs related to the comparison of L- and D-amino acids with respect to formation of amyloid fibrils and effect as molecular chaperones having therapeutic efficiencies.

DETAILED DESCRIPTION OF THE INVENTION

The present invention generally relates to therapeutic compositions for the treatment of mammalian disease and related methods. It more specifically relates to compositions comprising peptides for the treatment of inflammation and methods for delivering the peptides.

“Complex” refers to a molecular entity formed by non-covalent association involving two or more component molecular entities (ionic or uncharged). The bonding between the components is normally weaker than in a covalent bond. Typically, the dissociation constant for a complex (“K_d”) is equal to or greater than 1 μM. In certain cases, it is equal to or greater than “Fibril” refers to an aggregation of four or more peptides of the present invention. In some cases, the aggregate includes one hundred or more, one thousand or more, five thousand or more, or ten thousand or more peptides.

In a composition aspect, the present invention is directed to one or more peptides that exhibit a therapeutic effect when administered to a mammal, typically an anti-inflammatory effect. The one or more peptides comprise, or consist of, hexapeptides comprising amino acids of an L-configuration, D-configuration or a mixture of configurations that generate a free energy of formation of −23 kcal/mol or less (i.e., more negative) when the hexapeptide sequence in subjected to the Rosetta-Profile algorithm. In certain cases, the peptide is a hexapeptide. The peptides have a carboxy terminus and an amino terminus.

In certain cases the peptides of the present invention comprise 1, 2, or 3 amino acids having polar basic side chains. The polar basic side chains may have a terminal amino or a terminal imidazole group. In other cases, the peptides comprise 1 polar acidic side chain. In other cases, the peptides comprise 0, 1, 2, 3, 4, or 5 amino acids having hydrophobic side chains. In still other cases, the peptides comprise 0, 1, 2, 3, 4, or 5 amino acids having polar uncharged side chains, wherein the peptide has a positive charge.

The carboxy terminus of the peptides is typically either a carboxylic acid (i.e., —CO₂H) or an amide (i.e., —C(O)NR₂, where R is a substituent such as alkyl or hydrogen). The amino terminus is typically either an amine (i.e., N(R′)₂, where R′ is a substituent such as alkyl or hydrogen) or an acetate group (i.e., —C(O)R″, where R″ is a substituent such as methyl, ethyl, or longer alkyl).

The one or more peptides may comprise, or consist of, one or more of the following hexapeptides, where each indicated amino acid is either an L-amino acid or a D-amino acid (symbol for hexapeptide indicated in parenthesis after the hexamer):

SVNVDL;
(SEQ ID NO: 1)

SLNVDV;
(SEQ ID NO: 2)

SVDVNL;
(SEQ ID NO: 3)

DLSVVL;
(SEQ ID NO: 4)

SVNLDV;
(SEQ ID NO: 5)

SVVNDV;
(SEQ ID NO: 6)

DVSLVN;
(SEQ ID NO: 7)

DVSVLN;
(SEQ ID NO: 8)

SDLVNV;
(SEQ ID NO: 9)

SLNVVS;
(SEQ ID NO: 10)

LNVDSV;
(SEQ ID NO: 11)

NDLSVV;
(SEQ ID NO: 12)

VDNLVS;
(SEQ ID NO: 13)

VNDVSL;
(SEQ ID NO: 14)

VSDNVL;
(SEQ ID NO: 15)

LNDVVS;
(SEQ ID NO: 16)

LSVDVN;
(SEQ ID NO: 17)

NSVDLV;
(SEQ ID NO: 18)

VDVLNS;
(SEQ ID NO: 19)

VNDSVL;
(SEQ ID NO: 20)

VNVSLD;
(SEQ ID NO: 21)

LDVNSV;
(SEQ ID NO: 22)

LNVVDS;
(SEQ ID NO: 23)

LSDVVN;
(SEQ ID NO: 24)

LVVNDS;
(SEQ ID NO: 25)

NSLDVV;
(SEQ ID NO: 26)

VLVDNS;
(SEQ ID NO: 27)

VNLSDV;
(SEQ ID NO: 28)

VNVDLS;
(SEQ ID NO: 29)

DNVSVD;
(SEQ ID NO: 30)

LSDNVV;
(SEQ ID NO: 31)

LSVVDN;
(SEQ ID NO: 32)

VLDVSN;
(SEQ ID NO: 33)

VNDLVS;
(SEQ ID NO: 34)

VNVSDL;
(SEQ ID NO: 35)

VVLSDN;
(SEQ ID NO: 36)

LVSVNL;
(SEQ ID NO: 37)

LVNVSV;
(SEQ ID NO: 38)

SLNVSV;
(SEQ ID NO: 39)

LVSVNS;
(SEQ ID NO: 40)

SVDVNV;
(SEQ ID NO: 41)

LVVSVL;
(SEQ ID NO: 42)

VNLVVS;
(SEQ ID NO: 43)

SNLVSV;
(SEQ ID NO: 44)

SVNVLS;
(SEQ ID NO: 45)

VLVSVL;
(SEQ ID NO: 46)

LVNVSL;
(SEQ ID NO: 47)

SVNVDS;
(SEQ ID NO: 48)

VLSVNV;
(SEQ ID NO: 49)

NLVVSV;
(SEQ ID NO: 50)

SVVLNV;
(SEQ ID NO: 51)

LSVVNL;
(SEQ ID NO: 52)

SNLVVS;
(SEQ ID NO: 53)

SVVLDV;
(SEQ ID NO: 54)

LVSLNV;
(SEQ ID NO: 55)

VSLNVV;
(SEQ ID NO: 56)

VKVQIY;
(SEQ ID NO: 57)

QVVIYK;
(SEQ ID NO: 58)

KVIQVY;
(SEQ ID NO: 59)

VYVKIY;
(SEQ ID NO: 60)

QIVVYK;
(SEQ ID NO: 61)

QVIKVY;
(SEQ ID NO: 62)

VQVKIY;
(SEQ ID NO: 63)

QVVKIY;
(SEQ ID NO: 64)

QVIVYK;
(SEQ ID NO: 65)

QIVKVY;
(SEQ ID NO: 66)

QKIVVY;
(SEQ ID NO: 67)

QKVVYI;
(SEQ ID NO: 68)

KVQVYI;
(SEQ ID NO: 69)

QVVKYI;
(SEQ ID NO: 70)

KVQIYV;
(SEQ ID NO: 71)

VKIQVY;
(SEQ ID NO: 72)

VIQKVY;
(SEQ ID NO: 73)

KVVIYK;
(SEQ ID NO: 74)

QIVKYV;
(SEQ ID NO: 75)

QKVIYV;
(SEQ ID NO: 76)

KQVVIY;
(SEQ ID NO: 77)

KIQVYV;
(SEQ ID NO: 78)

KVYVQI;
(SEQ ID NO: 79)

VVQIYK;
(SEQ ID NO: 80)

KQVIVY;
(SEQ ID NO: 81)

VQIKVY;
(SEQ ID NO: 82)

QKIVYV;
(SEQ ID NO: 83)

VIQVYK;
(SEQ ID NO: 84)

KVVIQY;
(SEQ ID NO: 85)

KVQVIY;
(SEQ ID NO: 86)

QYVVIK;
(SEQ ID NO: 87)

YVVIQK;
(SEQ ID NO: 88)

KIVVQY;
(SEQ ID NO: 89)

YQVIVK;
(SEQ ID NO: 90)

KIYVQV;
(SEQ ID NO: 91)

KVVIYQ;
(SEQ ID NO: 92)

KYVVQI;
(SEQ ID NO: 93)

YQIVVK;
(SEQ ID NO: 94)

YVVIQY;
(SEQ ID NO: 95)

KQVIYV;
(SEQ ID NO: 96)

KYVQVI;
(SEQ ID NO: 97)

KYVVIQ;
(SEQ ID NO: 98)

QKVVIY;
(SEQ ID NO: 99)

QYVIVK;
(SEQ ID NO: 100)

IVQKVY;
(SEQ ID NO: 101)

KYIVVQ;
(SEQ ID NO: 102)

QVIKYV;
(SEQ ID NO: 103)

YIVVQK;
(SEQ ID NO: 104)

YQVVIK;
(SEQ ID NO: 105)

IKVQVY;
(SEQ ID NO: 106)

KVVQYI;
(SEQ ID NO: 107)

KYVQIV;
(SEQ ID NO: 108)

VKQVYI;
(SEQ ID NO: 109)

VQVIYK;
(SEQ ID NO: 110)

VVIQKY;
(SEQ ID NO: 111)

IQVVYK;
(SEQ ID NO: 112)

KIVVYQ;
(SEQ ID NO: 113)

KYQVIV;
(SEQ ID NO: 114)

KYVIVQ;
(SEQ ID NO: 115)

VVYIQK;
(SEQ ID NO: 116)

IVQVYK;
(SEQ ID NO: 117)

KQIVYV;
(SEQ ID NO: 118)

KYVIQV;
(SEQ ID NO: 119)

QKVIVY;
(SEQ ID NO: 120)

QYIKVV;
(SEQ ID NO: 121)

QYIVKV;
(SEQ ID NO: 122)

VQIVYK;
(SEQ ID NO: 123)

VVQKYI;
(SEQ ID NO: 124)

VYQIVK;
(SEQ ID NO: 125)

YVQVIK;
(SEQ ID NO: 126)

QIVYVK;
(SEQ ID NO: 127)

IKVYQV;
(SEQ ID NO: 128)

KVVYIQ;
(SEQ ID NO: 129)

VQKYIV;
(SEQ ID NO: 130)

VQKYVI;
(SEQ ID NO: 131)

KVVQIY;
(SEQ ID NO: 132)

YVIKIY;
(SEQ ID NO: 133)

QVIVYV;
(SEQ ID NO: 134)

VVIKVY;
(SEQ ID NO: 135)

KVVIYI;
(SEQ ID NO: 136)

VYVVIK;
(SEQ ID NO: 137)

YIVQVY;
(SEQ ID NO: 138)

YVIQVY;
(SEQ ID NO: 139)

YVIVVY;
(SEQ ID NO: 140)

KVIVYI;
(SEQ ID NO: 141)

YVIKVY;
(SEQ ID NO: 142)

VQVIVK;
(SEQ ID NO: 143)

YVVKIY;
(SEQ ID NO: 144)

YVVQVI;
(SEQ ID NO: 145)

QIVVYQ;
(SEQ ID NO: 146)

VVQIVK;
(SEQ ID NO: 147)

VQIVVK;
(SEQ ID NO: 148)

VIVVYK;
(SEQ ID NO: 149)

IVQVYI;
(SEQ ID NO: 150)

YVVIQV;
(SEQ ID NO: 151)

VYQVVI;
(SEQ ID NO: 152)

KYIQVY;
(SEQ ID NO: 153)

KYVVIY;
(SEQ ID NO: 154)

KYVQIY;
(SEQ ID NO: 155)

VVQVIK;
(SEQ ID NO: 156)

QKIVVK;
(SEQ ID NO: 157)

KYVIVY;
(SEQ ID NO: 158)

KQVVIK;
(SEQ ID NO: 159)

YVQIYV;
(SEQ ID NO: 160)

YKVQVY;
(SEQ ID NO: 161)

IKVQIY;
(SEQ ID NO: 162)

KIVVQK;
(SEQ ID NO: 163)

VIKVVI;
(SEQ ID NO: 164)

VYIKVV;
(SEQ ID NO: 165)

VYIVQV;
(SEQ ID NO: 166)

IVYVQI;
(SEQ ID NO: 167)

KVQIYK;
(SEQ ID NO: 168)

IYVIVY;
(SEQ ID NO: 169)

VIYVIV;
(SEQ ID NO: 170)

KYQVYI;
(SEQ ID NO: 171)

VIQKVV;
(SEQ ID NO: 172)

VKIVYV;
(SEQ ID NO: 173)

VKQIVV;
(SEQ ID NO: 174)

YIVKQY;
(SEQ ID NO: 175)

YIVVQY;
(SEQ ID NO: 176)

YKIQVY;
(SEQ ID NO: 177)

YQVVIY;
(SEQ ID NO: 178)

YVVKQY;
(SEQ ID NO: 179)

VIVKVQ;
(SEQ ID NO: 180)

QKVVIK;
(SEQ ID NO: 181)

GMVVVG;
(SEQ ID NO: 182)

GVVVMG;
(SEQ ID NO: 183)

GVVMVG;
(SEQ ID NO: 184)

GGVVVM;
(SEQ ID NO: 185)

GVVGVM;
(SEQ ID NO: 186)

GVVVGM;
(SEQ ID NO: 187)

GVMVVG;
(SEQ ID NO: 188)

GGVVMV;
(SEQ ID NO: 189)

GGVMVV;
(SEQ ID NO: 190)

GMVVGV;
(SEQ ID NO: 191)

GVVMGV;
(SEQ ID NO: 192)

GMVGVV;
(SEQ ID NO: 193)

GVMVGV;
(SEQ ID NO: 194)

GVVGMV;
(SEQ ID NO: 195)

GGMVVV;
(SEQ ID NO: 196)

GVGVMV;
(SEQ ID NO: 197)

GVGVVM;
(SEQ ID NO: 198)

MVVVGM;
(SEQ ID NO: 199)

GMGVVV;
(SEQ ID NO: 200)

MVVVGG;
(SEQ ID NO: 201)

GVGMVV;
(SEQ ID NO: 202)

GVVGGV;
(SEQ ID NO: 203)

MVGVVG;
(SEQ ID NO: 204)

VVMVGG;
(SEQ ID NO: 205)

MGVVVG;
(SEQ ID NO: 206)

MVGVGV;
(SEQ ID NO: 207)

VGVMVG;
(SEQ ID NO: 208)

MVVGVG;
(SEQ ID NO: 209)

VVGVMG;
(SEQ ID NO: 210)

VMGVVG;
(SEQ ID NO: 211)

VGVGVM;
(SEQ ID NO: 212)

VMVGVG;
(SEQ ID NO: 213)

MGGVVV;
(SEQ ID NO: 214)

VGMVVG;
(SEQ ID NO: 215)

VVGVGM;
(SEQ ID NO: 216)

VGGVVM;
(SEQ ID NO: 217)

VVGMVG;
(SEQ ID NO: 218)

VGVVMG;
(SEQ ID NO: 219)

MVGGVV;
(SEQ ID NO: 220)

VGMVGV;
(SEQ ID NO: 221)

VMVVGG;
(SEQ ID NO: 222)

VVGGVM;
(SEQ ID NO: 223)

VGGVMV;
(SEQ ID NO: 224)

VVVGMG;
(SEQ ID NO: 225)

VVGMGV;
(SEQ ID NO: 226)

MGVGVV;
(SEQ ID NO: 227)

VGVVGM;
(SEQ ID NO: 228)

VGGMVV;
(SEQ ID NO: 229)

VVVMGG;
(SEQ ID NO: 230)

VVMGGV;
(SEQ ID NO: 231)

VVVGGM;
(SEQ ID NO: 232)

VVGGMV;
(SEQ ID NO: 233)

VGMGVV;
(SEQ ID NO: 234)

VMGGVV;
(SEQ ID NO: 235)

VGGVGV;
(SEQ ID NO: 236)

VGVMGV;
(SEQ ID NO: 237)

VGVGMV;
(SEQ ID NO: 238)

and

VMVGGV;
(SEQ ID NO: 239)

ANSTSV;
(SEQ ID NO: 240)

ANSVSG;
(SEQ ID NO: 241)

ANSVSS;
(SEQ ID NO: 242)

AQNSNV;
(SEQ ID NO: 243)

AQNVNS;
(SEQ ID NO: 244)

AQNVTS;
(SEQ ID NO: 245)

AQSQSV;
(SEQ ID NO: 246)

AQSSSV;
(SEQ ID NO: 247)

AQSTSV;
(SEQ ID NO: 248)

AQSVNS;
(SEQ ID NO: 249)

AQSVQS;
(SEQ ID NO: 250)

AQSVSQ;
(SEQ ID NO: 251)

AQSVSS;
(SEQ ID NO: 252)

AQSVST;
(SEQ ID NO: 253)

ASNNNV;
(SEQ ID NO: 254)

ASNQNQ;
(SEQ ID NO: 255)

ASNQNV;
(SEQ ID NO: 256)

ASNQTQ;
(SEQ ID NO: 257)

ASNSNV;
(SEQ ID NO: 258)

ASNSTV;
(SEQ ID NO: 259)

ASNTNS;
(SEQ ID NO: 260)

ASNTNV;
(SEQ ID NO: 261)

ASNTSV;
(SEQ ID NO: 262)

ASNVNG;
(SEQ ID NO: 263)

ASNVNQ;
(SEQ ID NO: 264)

ASNVNS;
(SEQ ID NO: 265)

ASNVNT;
(SEQ ID NO: 266)

ASNVTG;
(SEQ ID NO: 267)

ASNVTT;
(SEQ ID NO: 268)

ASSNSV;
(SEQ ID NO: 269)

ASSVSG;
(SEQ ID NO: 270)

ASSVSN;
(SEQ ID NO: 271)

ATNVNS;
(SEQ ID NO: 272)

ATNVTS;
(SEQ ID NO: 273)

ATSQSQ;
(SEQ ID NO: 274)

ATSQSV;
(SEQ ID NO: 275)

ATSTSG;
(SEQ ID NO: 276)

ATSTSV;
(SEQ ID NO: 277)

ATSVSG;
(SEQ ID NO: 278)

ATSVSS;
(SEQ ID NO: 279)

AVNQNS;
(SEQ ID NO: 280)

AVNQSQ;
(SEQ ID NO: 281)

AVNSNG;
(SEQ ID NO: 282)

AVNSNS;
(SEQ ID NO: 283)

AVNSNT;
(SEQ ID NO: 284)

AVNTNS;
(SEQ ID NO: 285)

AVSNSG;
(SEQ ID NO: 286)

AVSNSS;
(SEQ ID NO: 287)

AVSQNQ;
(SEQ ID NO: 288)

AVSQSG;
(SEQ ID NO: 289)

AVSQSQ;
(SEQ ID NO: 290)

AVSQTQ;
(SEQ ID NO: 291)

AVSSNQ;
(SEQ ID NO: 292)

AVSSNS;
(SEQ ID NO: 293)

AVSSSQ;
(SEQ ID NO: 294)

AVSTSG;
(SEQ ID NO: 295)

GANTVS;
(SEQ ID NO: 296)

GAQTSS;
(SEQ ID NO: 297)

GASNQS;
(SEQ ID NO: 298)

GASQQS;
(SEQ ID NO: 299)

GASSQQ;
(SEQ ID NO: 300)

GGQVTS;
(SEQ ID NO: 301)

GGSNQV;
(SEQ ID NO: 302)

GNNVQS;
(SEQ ID NO: 303)

GNQVTS;
(SEQ ID NO: 304)

GNSNQV;
(SEQ ID NO: 305)

GNSQQQ;
(SEQ ID NO: 306)

GNSQQS;
(SEQ ID NO: 307)

GNSQQV;
(SEQ ID NO: 308)

GNSSTV;
(SEQ ID NO: 309)

GNSTQS;
(SEQ ID NO: 310)

GNSTQV;
(SEQ ID NO: 311)

GNSTVS;
(SEQ ID NO: 312)

GNSVQS;
(SEQ ID NO: 313)

GNSVSS;
(SEQ ID NO: 314)

GNSVST;
(SEQ ID NO: 315)

GNSVTS;
(SEQ ID NO: 316)

GQNTVS;
(SEQ ID NO: 317)

GQNVAS;
(SEQ ID NO: 318)

GQNVQS;
(SEQ ID NO: 319)

GQNVSS;
(SEQ ID NO: 320)

GQNVTS;
(SEQ ID NO: 321)

GQQQSQ;
(SEQ ID NO: 322)

GQQTSS;
(SEQ ID NO: 323)

GQQTSV;
(SEQ ID NO: 324)

GQQVAS;
(SEQ ID NO: 325)

GQQVNS;
(SEQ ID NO: 326)

GQQVQS;
(SEQ ID NO: 327)

GQQVSG;
(SEQ ID NO: 328)

GQQVSQ;
(SEQ ID NO: 329)

GQQVSS;
(SEQ ID NO: 330)

GQQVST;
(SEQ ID NO: 331)

GQQVTS;
(SEQ ID NO: 332)

GQSGQV;
(SEQ ID NO: 333)

GQSNQA;
(SEQ ID NO: 334)

GQSNQS;
(SEQ ID NO: 335)

GQSNQV;
(SEQ ID NO: 336)

GQSQAQ;
(SEQ ID NO: 337)

GQSQQQ;
(SEQ ID NO: 338)

GQSQQS;
(SEQ ID NO: 339)

GQSQSQ;
(SEQ ID NO: 340)

GQSSQQ;
(SEQ ID NO: 341)

GQSSQS;
(SEQ ID NO: 342)

GQSSQV;
(SEQ ID NO: 343)

GQSTQS;
(SEQ ID NO: 344)

GQSTQV;
(SEQ ID NO: 345)

GQSVAG;
(SEQ ID NO: 346)

GQSVAQ;
(SEQ ID NO: 347)

GQSVAS;
(SEQ ID NO: 348)

GQSVQN;
(SEQ ID NO: 349)

GQSVQQ;
(SEQ ID NO: 350)

GQSVQS;
(SEQ ID NO: 351)

GQSVSG;
(SEQ ID NO: 352)

GQSVSN;
(SEQ ID NO: 353)

GQSVSQ;
(SEQ ID NO: 354)

GQSVSS;
(SEQ ID NO: 355)

GQSVST;
(SEQ ID NO: 356)

GQSVTS;
(SEQ ID NO: 357)

GSNQQV;
(SEQ ID NO: 358)

GSNQVQ;
(SEQ ID NO: 359)

GSNSTV;
(SEQ ID NO: 360)

GSNSVQ;
(SEQ ID NO: 361)

GSNSVT;
(SEQ ID NO: 362)

GSNTAV;
(SEQ ID NO: 363)

GSNTQV;
(SEQ ID NO: 364)

GSNTVA;
(SEQ ID NO: 365)

GSNTVS;
(SEQ ID NO: 366)

GSNVAS;
(SEQ ID NO: 367)

GSNVQQ;
(SEQ ID NO: 368)

GSNVQS;
(SEQ ID NO: 369)

GSNVQT;
(SEQ ID NO: 370)

GSNVTS;
(SEQ ID NO: 371)

GSQQSV;
(SEQ ID NO: 372)

GSQQTQ;
(SEQ ID NO: 373)

GSQQTV;
(SEQ ID NO: 374)

GSQTSS;
(SEQ ID NO: 375)

GSQTSV;
(SEQ ID NO: 376)

GSQVAS;
(SEQ ID NO: 377)

GSQVNS;
(SEQ ID NO: 378)

GSQVQS;
(SEQ ID NO: 379)

GSQVSS;
(SEQ ID NO: 380)

GSQVST;
(SEQ ID NO: 381)

GSQVTG;
(SEQ ID NO: 382)

GSQVTS;
(SEQ ID NO: 383)

GSSAQS;
(SEQ ID NO: 384)

GSSGQV;
(SEQ ID NO: 385)

GSSNQA;
(SEQ ID NO: 386)

GSSNQT;
(SEQ ID NO: 387)

GSSNQV;
(SEQ ID NO: 388)

GSSNSV;
(SEQ ID NO: 389)

GSSNTV;
(SEQ ID NO: 390)

GSSNVT;
(SEQ ID NO: 391)

GSSQAQ;
(SEQ ID NO: 392)

GSSQQA;
(SEQ ID NO: 393)

GSSQQV;
(SEQ ID NO: 394)

GSSQSQ;
(SEQ ID NO: 395)

GSSSQQ;
(SEQ ID NO: 396)

GSSSQS;
(SEQ ID NO: 397)

GSSSQV;
(SEQ ID NO: 398)

GSSTQA;
(SEQ ID NO: 399)

GSSTQG;
(SEQ ID NO: 400)

GSSTQT;
(SEQ ID NO: 401)

GSSTQV;
(SEQ ID NO: 402)

GSSTSV;
(SEQ ID NO: 403)

GSSTVN;
(SEQ ID NO: 404)

GSSVNS;
(SEQ ID NO: 405)

GSSVQN;
(SEQ ID NO: 406)

GSSVQT;
(SEQ ID NO: 407)

GSSVSG;
(SEQ ID NO: 408)

GSSVSS;
(SEQ ID NO: 409)

GSSVST;
(SEQ ID NO: 410)

GSSVTN;
(SEQ ID NO: 411)

GTNSSV;
(SEQ ID NO: 412)

GTNSVS;
(SEQ ID NO: 413)

GTNTVS;
(SEQ ID NO: 414)

GTNVQS;
(SEQ ID NO: 415)

GTNVSS;
(SEQ ID NO: 416)

GTNVTS;
(SEQ ID NO: 417)

GTQQSQ;
(SEQ ID NO: 418)

GTQSTS;
(SEQ ID NO: 419)

GTQSTV;
(SEQ ID NO: 420)

GTQTSA;
(SEQ ID NO: 421)

GTQTSV;
(SEQ ID NO: 422)

GTQVQS;
(SEQ ID NO: 423)

GTQVSG;
(SEQ ID NO: 424)

GTQVSN;
(SEQ ID NO: 425)

GTQVSS;
(SEQ ID NO: 426)

GTQVST;
(SEQ ID NO: 427)

GTSAQS;
(SEQ ID NO: 428)

GTSGQV;
(SEQ ID NO: 429)

GTSNQA;
(SEQ ID NO: 430)

GTSNQS;
(SEQ ID NO: 431)

GTSNQT;
(SEQ ID NO: 432)

GTSNQV;
(SEQ ID NO: 433)

GTSNSV;
(SEQ ID NO: 434)

GTSNVS;
(SEQ ID NO: 435)

GTSQQA;
(SEQ ID NO: 436)

GTSQQS;
(SEQ ID NO: 437)

GTSQQV;
(SEQ ID NO: 438)

GTSQSQ;
(SEQ ID NO: 439)

GTSQSV;
(SEQ ID NO: 440)

GTSSNV;
(SEQ ID NO: 441)

GTSTQA;
(SEQ ID NO: 442)

GTSTQS;
(SEQ ID NO: 443)

GTSTQT;
(SEQ ID NO: 444)

GTSTQV;
(SEQ ID NO: 445)

GTSTSV;
(SEQ ID NO: 446)

GTSVAG;
(SEQ ID NO: 447)

GTSVAS;
(SEQ ID NO: 448)

GTSVNS;
(SEQ ID NO: 449)

GTSVNT;
(SEQ ID NO: 450)

GTSVQG;
(SEQ ID NO: 451)

GTSVQN;
(SEQ ID NO: 452)

GTSVQQ;
(SEQ ID NO: 453)

GTSVQS;
(SEQ ID NO: 454)

GTSVQT;
(SEQ ID NO: 455)

GTSVSG;
(SEQ ID NO: 456)

GTSVSN;
(SEQ ID NO: 457)

GTSVSS;
(SEQ ID NO: 458)

GTSVST;
(SEQ ID NO: 459)

GTSVTS;
(SEQ ID NO: 460)

GVNSQS;
(SEQ ID NO: 461)

GVNSST;
(SEQ ID NO: 462)

GVNSTS;
(SEQ ID NO: 463)

GVNTQS;
(SEQ ID NO: 464)

GVNTSS;
(SEQ ID NO: 465)

GVQNTS;
(SEQ ID NO: 466)

GVQQSQ;
(SEQ ID NO: 467)

GVQSNQ;
(SEQ ID NO: 468)

GVQSNS;
(SEQ ID NO: 469)

GVQSSG;
(SEQ ID NO: 470)

GVQSSQ;
(SEQ ID NO: 471)

GVQSTS;
(SEQ ID NO: 472)

GVQSTT;
(SEQ ID NO: 473)

GVQTNS;
(SEQ ID NO: 474)

GVQTQS;
(SEQ ID NO: 475)

GVQTSG;
(SEQ ID NO: 476)

GVQTSS;
(SEQ ID NO: 477)

GVSGQG;
(SEQ ID NO: 478)

GVSGQT;
(SEQ ID NO: 479)

GVSNAS;
(SEQ ID NO: 480)

GVSNNV;
(SEQ ID NO: 481)

GVSNQG;
(SEQ ID NO: 482)

GVSNQN;
(SEQ ID NO: 483)

GVSNQQ;
(SEQ ID NO: 484)

GVSNQS;
(SEQ ID NO: 485)

GVSNQT;
(SEQ ID NO: 486)

GVSNSS;
(SEQ ID NO: 487)

GVSNST;
(SEQ ID NO: 488)

GVSNTS;
(SEQ ID NO: 489)

GVSQAQ;
(SEQ ID NO: 490)

GVSQNQ;
(SEQ ID NO: 491)

GVSQNS;
(SEQ ID NO: 492)

GVSQQG;
(SEQ ID NO: 493)

GVSQQS;
(SEQ ID NO: 494)

GVSQQT;
(SEQ ID NO: 495)

GVSQSG;
(SEQ ID NO: 496)

GVSQSQ;
(SEQ ID NO: 497)

GVSQSS;
(SEQ ID NO: 498)

GVSSAQ;
(SEQ ID NO: 499)

GVSSAS;
(SEQ ID NO: 500)

GVSSGQ;
(SEQ ID NO: 501)

GVSSNG;
(SEQ ID NO: 502)

GVSSNQ;
(SEQ ID NO: 503)

GVSSNS;
(SEQ ID NO: 504)

GVSSNT;
(SEQ ID NO: 505)

GVSSQG;
(SEQ ID NO: 506)

GVSSQN;
(SEQ ID NO: 507)

GVSSQQ;
(SEQ ID NO: 508)

GVSSSG;
(SEQ ID NO: 509)

GVSSSQ;
(SEQ ID NO: 510)

GVSSSS;
(SEQ ID NO: 511)

GVSSTN;
(SEQ ID NO: 512)

GVSTAS;
(SEQ ID NO: 513)

GVSTNS;
(SEQ ID NO: 514)

GVSTQG;
(SEQ ID NO: 515)

GVSTQN;
(SEQ ID NO: 516)

GVSTQQ;
(SEQ ID NO: 517)

GVSTQS;
(SEQ ID NO: 518)

GVSTQT;
(SEQ ID NO: 519)

GVSTSN;
(SEQ ID NO: 520)

GVSTSQ;
(SEQ ID NO: 521)

GVSTSS;
(SEQ ID NO: 522)

GVSTST;
(SEQ ID NO: 523)

GVTNSS;
(SEQ ID NO: 524)

GVTSNS;
(SEQ ID NO: 525)

GVTSSN;
(SEQ ID NO: 526)

NGSTSV;
(SEQ ID NO: 527)

NGSVTS;
(SEQ ID NO: 528)

NNSVSS;
(SEQ ID NO: 529)

NQSQSQ;
(SEQ ID NO: 530)

NQSVSN;
(SEQ ID NO: 531)

NQSVSQ;
(SEQ ID NO: 532)

NQSVSS;
(SEQ ID NO: 533)

NSSNSV;
(SEQ ID NO: 534)

NSSQSQ;
(SEQ ID NO: 535)

NSSTVG;
(SEQ ID NO: 536)

NSSVSG;
(SEQ ID NO: 537)

NSSVSN;
(SEQ ID NO: 538)

NSSVSS;
(SEQ ID NO: 539)

NSSVTG;
(SEQ ID NO: 540)

NTNVNS;
(SEQ ID NO: 541)

NTQVSS;
(SEQ ID NO: 542)

NTSGSV;
(SEQ ID NO: 543)

NTSQSQ;
(SEQ ID NO: 544)

NTSTSS;
(SEQ ID NO: 545)

NTSTSV;
(SEQ ID NO: 546)

NTSVSS;
(SEQ ID NO: 547)

NVSGST;
(SEQ ID NO: 548)

NVSNSG;
(SEQ ID NO: 549)

NVSQSG;
(SEQ ID NO: 550)

NVSQSQ;
(SEQ ID NO: 551)

NVSQSS;
(SEQ ID NO: 552)

NVSSSG;
(SEQ ID NO: 553)

NVSSSS;
(SEQ ID NO: 554)

NVSSTG;
(SEQ ID NO: 555)

NVSTSN;
(SEQ ID NO: 556)

NVSTSS;
(SEQ ID NO: 557)

NVSTST;
(SEQ ID NO: 558)

QNSTSV;
(SEQ ID NO: 559)

QNSVSG;
(SEQ ID NO: 560)

QQSQSQ;
(SEQ ID NO: 561)

QQSVAQ;
(SEQ ID NO: 562)

QQSVAQ;
(SEQ ID NO: 563)

QQSVAS;
(SEQ ID NO: 564)

QQSVAT;
(SEQ ID NO: 565)

QQSVSG;
(SEQ ID NO: 566)

QQSVSN;
(SEQ ID NO: 567)

QQSVSS;
(SEQ ID NO: 568)

QSNQVQ;
(SEQ ID NO: 569)

QSNTAV;
(SEQ ID NO: 570)

QSNTNV;
(SEQ ID NO: 571)

QSSNSV;
(SEQ ID NO: 572)

QSSQAQ;
(SEQ ID NO: 573)

QSSQSQ;
(SEQ ID NO: 574)

QSSQSV;
(SEQ ID NO: 575)

QSSTSV;
(SEQ ID NO: 576)

QSSVSG;
(SEQ ID NO: 577)

QSSVSN;
(SEQ ID NO: 578)

QSSVSQ;
(SEQ ID NO: 579)

QSSVSS;
(SEQ ID NO: 580)

QSSVTG;
(SEQ ID NO: 581)

QSSVTS;
(SEQ ID NO: 582)

QTSQSQ;
(SEQ ID NO: 583)

QTSSSQ;
(SEQ ID NO: 584)

QTSSSV;
(SEQ ID NO: 585)

QTSVAG;
(SEQ ID NO: 586)

QTSVAS;
(SEQ ID NO: 587)

QTSVSG;
(SEQ ID NO: 588)

QTSVSN;
(SEQ ID NO: 589)

QTSVSS;
(SEQ ID NO: 590)

QTSVST;
(SEQ ID NO: 591)

QTSVTS;
(SEQ ID NO: 592)

QVSNSG;
(SEQ ID NO: 593)

QVSNSS;
(SEQ ID NO: 594)

QVSNST;
(SEQ ID NO: 595)

QVSQAQ;
(SEQ ID NO: 596)

QVSQTG;
(SEQ ID NO: 597)

QVSQTQ;
(SEQ ID NO: 598)

QVSSAQ;
(SEQ ID NO: 599)

QVSSSG;
(SEQ ID NO: 600)

QVSSSQ;
(SEQ ID NO: 601)

QVSSSS;
(SEQ ID NO: 602)

QVSSTQ;
(SEQ ID NO: 603)

QVSTSG;
(SEQ ID NO: 604)

QVSTSN;
(SEQ ID NO: 605)

QVSTSS;
(SEQ ID NO: 606)

QVSTST;
(SEQ ID NO: 607)

SAQQSQ;
(SEQ ID NO: 608)

SAQQTQ;
(SEQ ID NO: 609)

SASQQQ;
(SEQ ID NO: 610)

SASQSQ;
(SEQ ID NO: 611)

SGNSTV;
(SEQ ID NO: 612)

SGNTSV;
(SEQ ID NO: 613)

SGNVST;
(SEQ ID NO: 614)

SGNVTS;
(SEQ ID NO: 615)

SGQQTQ;
(SEQ ID NO: 616)

SGQQTV;
(SEQ ID NO: 617)

SGQTSV;
(SEQ ID NO: 618)

SGQVSS;
(SEQ ID NO: 619)

SGQVTG;
(SEQ ID NO: 620)

SGQVTQ;
(SEQ ID NO: 621)

SGQVTT;
(SEQ ID NO: 622)

SGSNTV;
(SEQ ID NO: 623)

SGSTNV;
(SEQ ID NO: 624)

SGSTQV;
(SEQ ID NO: 625)

SGSVAS;
(SEQ ID NO: 626)

SGSVNT;
(SEQ ID NO: 627)

SGSVQG;
(SEQ ID NO: 628)

SGSVQS;
(SEQ ID NO: 629)

SNNQTV;
(SEQ ID NO: 630)

SNNTAV;
(SEQ ID NO: 631)

SNNTNV;
(SEQ ID NO: 632)

SNNTQV;
(SEQ ID NO: 633)

SNQQSQ;
(SEQ ID NO: 634)

SNQQSV;
(SEQ ID NO: 635)

SNQQTQ;
(SEQ ID NO: 636)

SNQQTV;
(SEQ ID NO: 637)

SNQSTV;
(SEQ ID NO: 638)

SNQTSV;
(SEQ ID NO: 639)

SNQVSG;
(SEQ ID NO: 640)

SNQVSQ;
(SEQ ID NO: 641)

SNQVSS;
(SEQ ID NO: 642)

SNQVST;
(SEQ ID NO: 643)

SNQVTG;
(SEQ ID NO: 644)

SNQVTQ;
(SEQ ID NO: 645)

SNQVTS;
(SEQ ID NO: 646)

SNSGAV;
(SEQ ID NO: 647)

SNSNAV;
(SEQ ID NO: 648)

SNSNGV;
(SEQ ID NO: 649)

SNSNQS;
(SEQ ID NO: 650)

SNSQAG;
(SEQ ID NO: 651)

SNSQAQ;
(SEQ ID NO: 652)

SNSQAS;
(SEQ ID NO: 653)

SNSQAT;
(SEQ ID NO: 654)

SNSQAV;
(SEQ ID NO: 655)

SNSQGQ;
(SEQ ID NO: 656)

SNSQGV;
(SEQ ID NO: 657)

SNSQQA;
(SEQ ID NO: 658)

SNSQQG;
(SEQ ID NO: 659)

SNSQQQ;
(SEQ ID NO: 660)

SNSQQS;
(SEQ ID NO: 661)

SNSQQT;
(SEQ ID NO: 662)

SNSQQV;
(SEQ ID NO: 663)

SNSQSG;
(SEQ ID NO: 664)

SNSQST;
(SEQ ID NO: 665)

SNSQSV;
(SEQ ID NO: 666)

SNSSAV;
(SEQ ID NO: 667)

SNSSGV;
(SEQ ID NO: 668)

SNSSQG;
(SEQ ID NO: 669)

SNSSQQ;
(SEQ ID NO: 670)

SNSSQT;
(SEQ ID NO: 671)

SNSSQV;
(SEQ ID NO: 672)

SNSSSQ;
(SEQ ID NO: 673)

SNSSSV;
(SEQ ID NO: 674)

SNSTAG;
(SEQ ID NO: 675)

SNSTGS;
(SEQ ID NO: 676)

SNSTNV;
(SEQ ID NO: 677)

SNSTQA;
(SEQ ID NO: 678)

SNSTQQ;
(SEQ ID NO: 679)

SNSTQS;
(SEQ ID NO: 680)

SNSTQV;
(SEQ ID NO: 681)

SNSTSA;
(SEQ ID NO: 682)

SNSTSV;
(SEQ ID NO: 683)

SNSTVG;
(SEQ ID NO: 684)

SNSVAQ;
(SEQ ID NO: 685)

SNSVAS;
(SEQ ID NO: 686)

SNSVAT;
(SEQ ID NO: 687)

SNSVGQ;
(SEQ ID NO: 688)

SNSVGS;
(SEQ ID NO: 689)

SNSVGT;
(SEQ ID NO: 690)

SNSVNS;
(SEQ ID NO: 691)

SNSVNT;
(SEQ ID NO: 692)

SNSVQQ;
(SEQ ID NO: 693)

SNSVQS;
(SEQ ID NO: 694)

SNSVSG;
(SEQ ID NO: 695)

SNSVSQ;
(SEQ ID NO: 696)

SNSVSS;
(SEQ ID NO: 697)

SNSVST;
(SEQ ID NO: 698)

SNSVTG;
(SEQ ID NO: 699)

SQNVAS;
(SEQ ID NO: 700)

SQNVNS;
(SEQ ID NO: 701)

SQQQSQ;
(SEQ ID NO: 702)

SQQQSV;
(SEQ ID NO: 703)

SQQQTQ;
(SEQ ID NO: 704)

SQQQTV;
(SEQ ID NO: 705)

SQQSTV;
(SEQ ID NO: 706)

SQQVSG;
(SEQ ID NO: 707)

SQQVSN;
(SEQ ID NO: 708)

SQQVSS;
(SEQ ID NO: 709)

SQQVST;
(SEQ ID NO: 710)

SQQVTG;
(SEQ ID NO: 711)

SQQVTN;
(SEQ ID NO: 712)

SQQVTS;
(SEQ ID NO: 713)

SQQVTT;
(SEQ ID NO: 714)

SQSNAS;
(SEQ ID NO: 715)

SQSNAV;
(SEQ ID NO: 716)

SQSNGV;
(SEQ ID NO: 717)

SQSNQT;
(SEQ ID NO: 718)

SQSNQV;
(SEQ ID NO: 719)

SQSNSV;
(SEQ ID NO: 720)

SQSQAQ;
(SEQ ID NO: 721)

SQSQAS;
(SEQ ID NO: 722)

SQSQAV;
(SEQ ID NO: 723)

SQSQQQ;
(SEQ ID NO: 724)

SQSQQV;
(SEQ ID NO: 725)

SQSQSQ;
(SEQ ID NO: 726)

SQSSAV;
(SEQ ID NO: 727)

SQSSGV;
(SEQ ID NO: 728)

SQSSQG;
(SEQ ID NO: 729)

SQSSQV;
(SEQ ID NO: 730)

SQSSSQ;
(SEQ ID NO: 731)

SQSSSV;
(SEQ ID NO: 732)

SQSTAV;
(SEQ ID NO: 733)

SQSTQS;
(SEQ ID NO: 734)

SQSTSG;
(SEQ ID NO: 735)

SQSTSV;
(SEQ ID NO: 736)

SQSVAG;
(SEQ ID NO: 737)

SQSVAN;
(SEQ ID NO: 738)

SQSVAQ;
(SEQ ID NO: 739)

SQSVAS;
(SEQ ID NO: 740)

SQSVAT;
(SEQ ID NO: 741)

SQSVGG;
(SEQ ID NO: 742)

SQSVGN;
(SEQ ID NO: 743)

SQSVGQ;
(SEQ ID NO: 744)

SQSVGS;
(SEQ ID NO: 745)

SQSVGT;
(SEQ ID NO: 746)

SQSVNG;
(SEQ ID NO: 747)

SQSVNS;
(SEQ ID NO: 748)

SQSVQG;
(SEQ ID NO: 749)

SQSVQN;
(SEQ ID NO: 750)

SQSVQQ;
(SEQ ID NO: 751)

SQSVQS;
(SEQ ID NO: 752)

SQSVQT;
(SEQ ID NO: 753)

SQSVSG;
(SEQ ID NO: 754)

SQSVSN;
(SEQ ID NO: 755)

SQSVSS;
(SEQ ID NO: 756)

SQSVST;
(SEQ ID NO: 757)

SSNGTV;
(SEQ ID NO: 758)

SSNQNQ;
(SEQ ID NO: 759)

SSNQNV;
(SEQ ID NO: 760)

SSNQTQ;
(SEQ ID NO: 761)

SSNQTV;
(SEQ ID NO: 762)

SSNSTV;
(SEQ ID NO: 763)

SSNTNV;
(SEQ ID NO: 764)

SSNTQV;
(SEQ ID NO: 765)

SSNTVG;
(SEQ ID NO: 766)

SSNVAQ;
(SEQ ID NO: 767)

SSNVAS;
(SEQ ID NO: 768)

SSNVGT;
(SEQ ID NO: 769)

SSNVNG;
(SEQ ID NO: 770)

SSNVNQ;
(SEQ ID NO: 771)

SSNVNS;
(SEQ ID NO: 772)

SSNVNT;
(SEQ ID NO: 773)

SSNVQS;
(SEQ ID NO: 774)

SSNVQT;
(SEQ ID NO: 775)

SSNVTG;
(SEQ ID NO: 776)

SSNVTQ;
(SEQ ID NO: 777)

SSNVTS;
(SEQ ID NO: 778)

SSQNTV;
(SEQ ID NO: 779)

SSQQSV;
(SEQ ID NO: 780)

SSQQTQ;
(SEQ ID NO: 781)

SSQQTV;
(SEQ ID NO: 782)

SSQSTQ;
(SEQ ID NO: 783)

SSQTSV;
(SEQ ID NO: 784)

SSQVNS;
(SEQ ID NO: 785)

SSQVSN;
(SEQ ID NO: 786)

SSQVSQ;
(SEQ ID NO: 787)

SSQVSS;
(SEQ ID NO: 788)

SSQVST;
(SEQ ID NO: 789)

SSQVTG;
(SEQ ID NO: 790)

SSQVTQ;
(SEQ ID NO: 791)

SSQVTS;
(SEQ ID NO: 792)

SSQVTT;
(SEQ ID NO: 793)

SSSNAV;
(SEQ ID NO: 794)

SSSNQV;
(SEQ ID NO: 795)

SSSNSV;
(SEQ ID NO: 796)

SSSQAQ;
(SEQ ID NO: 797)

SSSQNQ;
(SEQ ID NO: 798)

SSSQQQ;
(SEQ ID NO: 799)

SSSQQT;
(SEQ ID NO: 800)

SSSQQV;
(SEQ ID NO: 801)

SSSQSQ;
(SEQ ID NO: 802)

SSSQSV;
(SEQ ID NO: 803)

SSSSGQ;
(SEQ ID NO: 804)

SSSSGV;
(SEQ ID NO: 805)

SSSSNV;
(SEQ ID NO: 806)

SSSSQQ;
(SEQ ID NO: 807)

SSSSQS;
(SEQ ID NO: 808)

SSSTAV;
(SEQ ID NO: 809)

SSSTNV;
(SEQ ID NO: 810)

SSSTSV;
(SEQ ID NO: 811)

SSSVAQ;
(SEQ ID NO: 812)

SSSVAS;
(SEQ ID NO: 813)

SSSVGG;
(SEQ ID NO: 814)

SSSVGQ;
(SEQ ID NO: 815)

SSSVGS;
(SEQ ID NO: 816)

SSSVNG;
(SEQ ID NO: 817)

SSSVNS;
(SEQ ID NO: 818)

SSSVNT;
(SEQ ID NO: 819)

SSSVQG;
(SEQ ID NO: 820)

SSSVQN;
(SEQ ID NO: 821)

SSSVQS;
(SEQ ID NO: 822)

SSSVQT;
(SEQ ID NO: 823)

SSSVSG;
(SEQ ID NO: 824)

SSSVSN;
(SEQ ID NO: 825)

SSSVSQ;
(SEQ ID NO: 826)

SSSVSS;
(SEQ ID NO: 827)

SSSVST;
(SEQ ID NO: 828)

STNGSV;
(SEQ ID NO: 829)

STNSGV;
(SEQ ID NO: 830)

STNSNV;
(SEQ ID NO: 831)

STNTNV;
(SEQ ID NO: 832)

STNTQV;
(SEQ ID NO: 833)

STNVGS;
(SEQ ID NO: 834)

STNVNG;
(SEQ ID NO: 835)

STNVNS;
(SEQ ID NO: 836)

STNVSG;
(SEQ ID NO: 837)

STNVSS;
(SEQ ID NO: 838)

STNVTG;
(SEQ ID NO: 839)

STNVTS;
(SEQ ID NO: 840)

STQQSG;
(SEQ ID NO: 841)

STQQSQ;
(SEQ ID NO: 842)

STQQSV;
(SEQ ID NO: 843)

STQQTQ;
(SEQ ID NO: 844)

STQQTS;
(SEQ ID NO: 845)

STQQTV;
(SEQ ID NO: 846)

STQSNV;
(SEQ ID NO: 847)

STQSTV;
(SEQ ID NO: 848)

STQTSS;
(SEQ ID NO: 849)

STQTSV;
(SEQ ID NO: 850)

STQVNS;
(SEQ ID NO: 851)

STQVSG;
(SEQ ID NO: 852)

STQVSN;
(SEQ ID NO: 853)

STQVST;
(SEQ ID NO: 854)

STQVTG;
(SEQ ID NO: 855)

STQVTN;
(SEQ ID NO: 856)

STQVTQ;
(SEQ ID NO: 857)

STQVTS;
(SEQ ID NO: 858)

STQVTT;
(SEQ ID NO: 859)

STSGNV;
(SEQ ID NO: 860)

STSNAS;
(SEQ ID NO: 861)

STSNGV;
(SEQ ID NO: 862)

STSNQV;
(SEQ ID NO: 863)

STSNSV;
(SEQ ID NO: 864)

STSQAQ;
(SEQ ID NO: 865)

STSQGV;
(SEQ ID NO: 866)

STSQNQ;
(SEQ ID NO: 867)

STSQQQ;
(SEQ ID NO: 868)

STSQQV;
(SEQ ID NO: 869)

STSQSQ;
(SEQ ID NO: 870)

STSSAV;
(SEQ ID NO: 871)

STSSGV;
(SEQ ID NO: 872)

STSSQQ;
(SEQ ID NO: 873)

STSSQS;
(SEQ ID NO: 874)

STSSQV;
(SEQ ID NO: 875)

STSSSQ;
(SEQ ID NO: 876)

STSTAV;
(SEQ ID NO: 877)

STSTGV;
(SEQ ID NO: 878)

STSTQA;
(SEQ ID NO: 879)

STSTQG;
(SEQ ID NO: 880)

STSTQT;
(SEQ ID NO: 881)

STSTQV;
(SEQ ID NO: 882)

STSTSQ;
(SEQ ID NO: 883)

STSTSV;
(SEQ ID NO: 884)

STSVAG;
(SEQ ID NO: 885)

STSVAN;
(SEQ ID NO: 886)

STSVAQ;
(SEQ ID NO: 887)

STSVAS;
(SEQ ID NO: 888)

STSVAT;
(SEQ ID NO: 889)

STSVGG;
(SEQ ID NO: 890)

STSVGN;
(SEQ ID NO: 891)

STSVGQ;
(SEQ ID NO: 892)

STSVGS;
(SEQ ID NO: 893)

STSVNG;
(SEQ ID NO: 894)

STSVNN;
(SEQ ID NO: 895)

STSVNS;
(SEQ ID NO: 896)

STSVQG;
(SEQ ID NO: 897)

STSVQN;
(SEQ ID NO: 898)

STSVQQ;
(SEQ ID NO: 899)

STSVQS;
(SEQ ID NO: 900)

STSVQT;
(SEQ ID NO: 901)

STSVSG;
(SEQ ID NO: 902)

STSVSN;
(SEQ ID NO: 903)

STSVSQ;
(SEQ ID NO: 904)

STSVSS;
(SEQ ID NO: 905)

STSVST;
(SEQ ID NO: 906)

SVNGST;
(SEQ ID NO: 907)

SVNGTS;
(SEQ ID NO: 908)

SVNQAQ;
(SEQ ID NO: 909)

SVNQQQ;
(SEQ ID NO: 910)

SVNSGT;
(SEQ ID NO: 911)

SVNSNQ;
(SEQ ID NO: 912)

SVNSNS;
(SEQ ID NO: 913)

SVNSTG;
(SEQ ID NO: 914)

SVNSTS;
(SEQ ID NO: 915)

SVNTGS;
(SEQ ID NO: 916)

SVNTSG;
(SEQ ID NO: 917)

SVQQSQ;
(SEQ ID NO: 918)

SVQQST;
(SEQ ID NO: 919)

SVQQTQ;
(SEQ ID NO: 920)

SVQQTS;
(SEQ ID NO: 921)

SVQQTT;
(SEQ ID NO: 922)

SVQSNQ;
(SEQ ID NO: 923)

SVQSNS;
(SEQ ID NO: 924)

SVQSSG;
(SEQ ID NO: 925)

SVQSSQ;
(SEQ ID NO: 926)

SVQSSS;
(SEQ ID NO: 927)

SVQSTG;
(SEQ ID NO: 928)

SVQSTQ;
(SEQ ID NO: 929)

SVQSTS;
(SEQ ID NO: 930)

SVQTSG;
(SEQ ID NO: 931)

SVQTSN;
(SEQ ID NO: 932)

SVQTSS;
(SEQ ID NO: 933)

SVQVSN;
(SEQ ID NO: 934)

SVSGNT;
(SEQ ID NO: 935)

SVSGQS;
(SEQ ID NO: 936)

SVSNAQ;
(SEQ ID NO: 937)

SVSNAS;
(SEQ ID NO: 938)

SVSNGS;
(SEQ ID NO: 939)

SVSNGT;
(SEQ ID NO: 940)

SVSNQG;
(SEQ ID NO: 941)

SVSNQS;
(SEQ ID NO: 942)

SVSNQT;
(SEQ ID NO: 943)

SVSNST;
(SEQ ID NO: 944)

SVSNTG;
(SEQ ID NO: 945)

SVSQAQ;
(SEQ ID NO: 946)

SVSQAS;
(SEQ ID NO: 947)

SVSQGQ;
(SEQ ID NO: 948)

SVSQNQ;
(SEQ ID NO: 949)

SVSQQG;
(SEQ ID NO: 950)

SVSQQQ;
(SEQ ID NO: 951)

SVSQQS;
(SEQ ID NO: 952)

SVSQSG;
(SEQ ID NO: 953)

SVSQSS;
(SEQ ID NO: 954)

SVSQTQ;
(SEQ ID NO: 955)

SVSSAQ;
(SEQ ID NO: 956)

SVSSAS;
(SEQ ID NO: 957)

SVSSGG;
(SEQ ID NO: 958)

SVSSGS;
(SEQ ID NO: 959)

SVSSGT;
(SEQ ID NO: 960)

SVSSNG;
(SEQ ID NO: 961)

SVSSNQ;
(SEQ ID NO: 962)

SVSSNS;
(SEQ ID NO: 963)

SVSSNT;
(SEQ ID NO: 964)

SVSSQG;
(SEQ ID NO: 965)

SVSSQQ;
(SEQ ID NO: 966)

SVSSQS;
(SEQ ID NO: 967)

SVSSQT;
(SEQ ID NO: 968)

SVSSQV;
(SEQ ID NO: 969)

SVSSSG;
(SEQ ID NO: 970)

SVSSSQ;
(SEQ ID NO: 971)

SVSSSS;
(SEQ ID NO: 972)

SVSTAG;
(SEQ ID NO: 973)

SVSTAS;
(SEQ ID NO: 974)

SVSTAT;
(SEQ ID NO: 975)

SVSTGS;
(SEQ ID NO: 976)

SVSTNS;
(SEQ ID NO: 977)

SVSTNT;
(SEQ ID NO: 978)

SVSTQQ;
(SEQ ID NO: 979)

SVSTQS;
(SEQ ID NO: 980)

SVSTQT;
(SEQ ID NO: 981)

SVSTSG;
(SEQ ID NO: 982)

SVSTSN;
(SEQ ID NO: 983)

SVSTSS;
(SEQ ID NO: 984)

SVSTST;
(SEQ ID NO: 985)

TASQSQ;
(SEQ ID NO: 986)

TASQVQ;
(SEQ ID NO: 987)

TGSNSV;
(SEQ ID NO: 988)

TGSNVS;
(SEQ ID NO: 989)

TGSSNV;
(SEQ ID NO: 990)

TGSVNS;
(SEQ ID NO: 991)

TGSVSN;
(SEQ ID NO: 992)

TNSGSV;
(SEQ ID NO: 993)

TNSGVS;
(SEQ ID NO: 994)

TNSQVQ;
(SEQ ID NO: 995)

TNSSGV;
(SEQ ID NO: 996)

TNSVGS;
(SEQ ID NO: 997)

TNSVSG;
(SEQ ID NO: 998)

TQSQSQ;
(SEQ ID NO: 999)

TQSQVQ;
(SEQ ID NO: 1000)

TQSTVS;
(SEQ ID NO: 1001)

TQSVSN;
(SEQ ID NO: 1002)

TQSVSS;
(SEQ ID NO: 1003)

TSNGVS;
(SEQ ID NO: 1004)

TSNVGS;
(SEQ ID NO: 1005)

TSNVSG;
(SEQ ID NO: 1006)

TSSGNV;
(SEQ ID NO: 1007)

TSSGVN;
(SEQ ID NO: 1008)

TSSNGV;
(SEQ ID NO: 1009)

TSSNSV;
(SEQ ID NO: 1010)

TSSNVG;
(SEQ ID NO: 1011)

TSSQSQ;
(SEQ ID NO: 1012)

TSSSVQ;
(SEQ ID NO: 1013)

TSSVGN;
(SEQ ID NO: 1014)

TSSVNG;
(SEQ ID NO: 1015)

TSSVSG;
(SEQ ID NO: 1016)

TSSVSS;
(SEQ ID NO: 1017)

TSSVTS;
(SEQ ID NO: 1018)

TVNSGS;
(SEQ ID NO: 1019)

TVNSSG;
(SEQ ID NO: 1020)

TVSGNS;
(SEQ ID NO: 1021)

TVSGSN;
(SEQ ID NO: 1022)

TVSNGS;
(SEQ ID NO: 1023)

TVSNSG;
(SEQ ID NO: 1024)

TVSNST;
(SEQ ID NO: 1025)

TVSNVG;
(SEQ ID NO: 1026)

TVSQNQ;
(SEQ ID NO: 1027)

TVSQQV;
(SEQ ID NO: 1028)

TVSQSG;
(SEQ ID NO: 1029)

TVSQSQ;
(SEQ ID NO: 1030)

TVSSGN;
(SEQ ID NO: 1031)

TVSSNG;
(SEQ ID NO: 1032)

TVSSNQ;
(SEQ ID NO: 1033)

TVSSNS;
(SEQ ID NO: 1034)

TVSSSQ;
(SEQ ID NO: 1035)

TVSTSG;
(SEQ ID NO: 1036)

TVSTSN;
(SEQ ID NO: 1037)

TVSTSS;
(SEQ ID NO: 1038)

TVSTST;
(SEQ ID NO: 1039)

VGSTNS;
(SEQ ID NO: 1040)

VNSTSG;
(SEQ ID NO: 1041)

VNSTSN;
(SEQ ID NO: 1042)

VSSQSQ;
(SEQ ID NO: 1043)

VSSQVQ;
(SEQ ID NO: 1044)

VSSTNG;
(SEQ ID NO: 1045)

VTSNSG;
(SEQ ID NO: 1046)

and

VTSQSQ.
(SEQ ID NO: 1047)

Oftentimes, the peptide comprises one or more of the following hexapeptides, where each indicated amino acid is either an L-amino acid or a D-amino acid:

SVNLDV
(SEQ ID NO: 5);

VEALYL
(SEQ ID NO: 1048);

LYQLEN
(SEQ ID NO: 1049);

VQIVYK
(SEQ ID NO: 123);

GYVIIK
(SEQ ID NO: 1050);

SNQNNF
(SEQ ID NO: 1051);

SSQVTQ
(SEQ ID NO: 791);

SVLTSL
(SEQ ID NO: 1052);

SSTNVG
(SEQ ID NO: 1053);

SVSSSY
(SEQ ID NO: 1054);

GAILSS
(SEQ ID NO: 1055);

GAIIGL
(SEQ ID NO: 1056);

AIIGLM
(SEQ ID NO: 1057);

MVGGVV
(SEQ ID NO: 220);

GGVVIA
(SEQ ID NO: 1058).

Peptides of the present invention are capable of forming fibrils. The corresponding fibrils are typically capable of forming a complex with one or more plasma proteins. Nonlimiting examples of such plasma proteins include (symbol for plasma protein indicated in parenthesis after the plasma protein): Apolipoprotein B-100 (A); Complement C3 (B); Complement C1s (C); Beta-2-glycoprotein 1 (D); Clusterin (E); Coagulation Factor V (F); Complement C1r (G); Apolipoprotein A-I (H); ITIH2 (I); Complement 1qB (J); Apolipoprotein A-IV (K); Complement Factor H (L); Fibrinogen beta chain (M); Complement C4-A (N); Transthyretin (0); SerpinG1 Plasma protease C1 inhibitor (P); Fibrinogen alpha chain (Q); Complement C1qA (R); Vitronectin (S); Serpina-1 Alpha-1-antitrypsin (T); VitaminD-binding protein (U); Haptoglobin-related protein (V); ITIH4 (W); Fibrinogen gamma chain (X); SerpinC1 Antithrombin-III (Y); Apolipoprotein A-2 (Z); Complement Factor H-related protein (AA); Gelsolin (BB); Complement factor B (CC); Alpha-2-HS-glycoprotein (DD); Serum paraoxinase/arylesterase 1 (EE); Complement C5 (FF); Apolipoprotein C5 (GG); Apolipoprotein C-II (HH); Apolipoprotein C-I (II); ITIH1 (JJ); von Willebrand factor (KK); Ceruloplasmin (LL); Apolipoprotein E (MM); Filamin-A (NN); Histidine-rich glycoprotein (00); SerpinF2 alpha-2-antiplasmin (PP); Coagulation factor II Prothrombin (QQ); Coagulation factor X (RR); Vitamin K-dependent protein (SS); Apolipoprotein C-III (TT); Alpha-1-acid glycoprotein 2 (UU); Coagulation factor IX (VV); Apolipoprotein M (WW); Serum Albumin (XX).

Examples of peptide fibril/plasma protein complexes that are capable of being formed include (symbol for hexamer followed by symbol for plasma protein):

1B, 2B, 3B, 4B, 5B, 6B, 7B, 8B, 9B, 10B, 11B, 12B, 13B, 14B, 15B, 16B, 17B, 18B, 19B, 20B, 21B, 22B, 23B, 24B, 25B, 26B, 27B, 28B, 29B, 30B, 31B, 32B, 33B, 34B, 35B, 36B, 37B, 38B, 39B, 40B, 41B, 42B, 43B, 44B, 45B, 46B, 47B, 48B, 49B, 50B, 51B, 52B, 53B, 54B, 55B, 56B, 57B, 58B, 59B, 60B, 61B, 62B, 63B, 64B, 65B, 66B, 67B, 68B, 69B, 70B, 71B, 72B, 73B, 74B, 75B, 76B, 77B, 78B, 79B, 80B, 81B, 82B, 83B, 84B, 85B, 86B, 87B, 88B, 89B, 90B, 91B, 92B, 93B, 94B, 95B, 96B, 97B, 98B, 99B, 100B, 101B, 102B, 103B, 104B, 105B, 106B, 107B, 108B, 109B, 110B, 111B, 112B, 113B, 114B, 115B, 116B, 117B, 118B, 119B, 120B, 121B, 122B, 123B, 124B, 125B, 126B, 127B, 128B, 129B, 130B, 131B, 132B, 133B, 134B, 135B, 136B, 137B, 138B, 139B, 140B, 141B, 142B, 143B, 144B, 145B, 146B, 147B, 148B, 149B, 150B, 151B, 152B, 153B, 154B, 155B, 156B, 157B, 158B, 159B, 160B, 161B, 162B, 163B, 164B, 165B, 166B, 167B, 168B, 169B, 170B, 171B, 172B, 173B, 174B, 175B, 176B, 177B, 178B, 179B, 180B, 181B, 182B, 183B, 184B, 185B, 186B, 187B, 188B, 189B, 190B, 191B, 192B, 193B, 194B, 195B, 196B, 197B, 198B, 199B, 200B, 201B, 202B, 203B, 204B, 205B, 206B, 207B, 208B, 209B, 210B, 211B, 212B, 213B, 214B, 215B, 216B, 217B, 218B, 219B, 220B, 221B, 222B, 223B, 224B, 225B, 226B, 227B, 228B, 229B, 230B, 231B, 232B, 233B, 234B, 235B, 236B, 237B, 238B, 239B, 240B, 241B, 242B, 243B, 244B, 245B, 246B; and 247B-1047B;

1C, 2C, 3C, 4C, 5C, 6C, 7C, 8C, 9C, 10C, 11C, 12C, 13C, 14C, 15C, 16C, 17C, 18C, 19C, 20C, 21C, 22C, 23C, 24C, 25C, 26C, 27C, 28C, 29C, 30C, 31C, 32C, 33C, 34C, 35C, 36C, 37C, 38C, 39C, 40C, 41C, 42C, 43C, 44C, 45C, 46C, 47C, 48C, 49C, 50C, 51C, 52C, 53C, 54C, 55C, 56C, 57C, 58C, 59C, 60C, 61C, 62C, 63C, 64C, 65C, 66C, 67C, 68C, 69C, 70C, 71C, 72C, 73C, 74C, 75C, 76C, 77C, 78C, 79C, 80C, 81C, 82C, 83C, 84C, 85C, 86C, 87C, 88C, 89C, 90C, 91C, 92C, 93C, 94C, 95C, 96C, 97C, 98C, 99C, 100C, 101C, 102C, 103C, 104C, 105C, 106C, 107C, 108C, 109C, 110C, 111C, 112C, 113C, 114C, 115C, 116C, 117C, 118C, 119C, 120C, 121C, 122C, 123C, 124C, 125C, 126C, 127C, 128C, 129C, 130C, 131C, 132C, 133C, 134C, 135C, 136C, 137C, 138C, 139C, 140C, 141C, 142C, 143C, 144C, 145C, 146C, 147C, 148C, 149C, 150C, 151C, 152C, 153C, 154C, 155C, 156C, 157C, 158C, 159C, 160C, 161C, 162C, 163C, 164C, 165C, 166C, 167C, 168C, 169C, 170C, 171C, 172C, 173C, 174C, 175C, 176C, 177C, 178C, 179C, 180C, 181C, 182C, 183C, 184C, 185C, 186C, 187C, 188C, 189C, 190C, 191C, 192C, 193C, 194C, 195C, 196C, 197C, 198C, 199C, 200C, 201C, 202C, 203C, 204C, 205C, 206C, 207C, 208C, 209C, 210C, 211C, 212C, 213C, 214C, 215C, 216C, 217C, 218C, 219C, 220C, 221C, 222C, 223C, 224C, 225C, 226C, 227C, 228C, 229C, 230C, 231C, 232C, 233C, 234C, 235C, 236C, 237C, 238C, 239C, 240C, 241C, 242C, 243C, 244C, 245C, 246C; and 247C-1047C;

1F, 2F, 3F, 4F, 5F, 6F, 7F, 8F, 9F, 10F, 11F, 12F, 13F, 14F, 15F, 16F, 17F, 18F, 19F, 20F, 21F, 22F, 23F, 24F, 25F, 26F, 27F, 28F, 29F, 30F, 31F, 32F, 33F, 34F, 35F, 36F, 37F, 38F, 39F, 40F, 41F, 42F, 43F, 44F, 45F, 46F, 47F, 48F, 49F, 50F, 51F, 52F, 53F, 54F, 55F, 56F, 57F, 58F, 59F, 60F, 61F, 62F, 63F, 64F, 65F, 66F, 67F, 68F, 69F, 70F, 71F, 72F, 73F, 74F, 75F, 76F, 77F, 78F, 79F, 80F, 81F, 82F, 83F, 84F, 85F, 86F, 87F, 88F, 89F, 90F, 91F, 92F, 93F, 94F, 95F, 96F, 97F, 98F, 99F, 100F, 101F, 102F, 103F, 104F, 105F, 106F, 107F, 108F, 109F, 110F, 111F, 112F, 113F, 114F, 115F, 116F, 117F, 118F, 119F, 120F, 121F, 122F, 123F, 124F, 125F, 126F, 127F, 128F, 129F, 130F, 131F, 132F, 133F, 134F, 135F, 136F, 137F, 138F, 139F, 140F, 141F, 142F, 143F, 144F, 145F, 146F, 147F, 148F, 149F, 150F, 151F, 152F, 153F, 154F, 155F, 156F, 157F, 158F, 159F, 160F, 161F, 162F, 163F, 164F, 165F, 166F, 167F, 168F, 169F, 170F, 171F, 172F, 173F, 174F, 175F, 176F, 177F, 178F, 179F, 180F, 181F, 182F, 183F, 184F, 185F, 186F, 187F, 188F, 189F, 190F, 191F, 192F, 193F, 194F, 195F, 196F, 197F, 198F, 199F, 200F, 201F, 202F, 203F, 204F, 205F, 206F, 207F, 208F, 209F, 210F, 211F, 212F, 213F, 214F, 215F, 216F, 217F, 218F, 219F, 220F, 221F, 222F, 223F, 224F, 225F, 226F, 227F, 228F, 229F, 230F, 231F, 232F, 233F, 234F, 235F, 236F, 237F, 238F, 239F, 240F, 241F, 242F, 243F, 244F, 245F, 246F; and 247F-1047F;

1G, 2G, 3G, 4G, 5G, 6G, 7G, 8G, 9G, 10G, 11G, 12G, 13G, 14G, 15G, 16G, 17G, 18G, 19G, 20G, 21G, 22G, 23G, 24G, 25G, 26G, 27G, 28G, 29G, 30G, 31G, 32G, 33G, 34G, 35G, 36G, 37G, 38G, 39G, 40G, 41G, 42G, 43G, 44G, 45G, 46G, 47G, 48G, 49G, 50G, 51G, 52G, 53G, 54G, 55G, 56G, 57G, 58G, 59G, 60G, 61G, 62G, 63G, 64G, 65G, 66G, 67G, 68G, 69G, 70G, 71G, 72G, 73G, 74G, 75G, 76G, 77G, 78G, 79G, 80G, 81G, 82G, 83G, 84G, 85G, 86G, 87G, 88G, 89G, 90G, 91G, 92G, 93G, 94G, 95G, 96G, 97G, 98G, 99G, 100G, 101G, 102G, 103G, 104G, 105G, 106G, 107G, 108G, 1090, 110G, 1110, 112G, 113G, 114G, 115G, 116G, 117G, 118G, 119G, 120G, 121G, 122G, 123G, 124G, 125G, 126G, 127G, 128G, 129G, 130G, 131G, 1320, 133G, 134G, 135G, 136G, 137G, 138G, 139G, 140G, 141G, 142G, 143G, 144G, 145G, 146G, 147G, 148G, 149G, 150G, 151G, 152G, 153G, 154G, 155G, 156G, 157G, 158G, 159G, 160G, 161G, 162G, 163G, 164G, 165G, 166G, 167G, 168G, 169G, 170G, 171G, 172G, 173G, 1740, 175G, 176G, 177G, 178G, 179G, 180G, 181G, 182G, 183G, 184G, 185G, 186G, 187G, 188G, 189G, 190G, 191G, 192G, 193G, 194G, 195G, 196G, 197G, 198G, 199G, 200G, 201G, 202G, 203G, 204G, 205G, 206G, 207G, 208G, 209G, 210G, 211G, 212G, 213G, 214G, 215G, 216G, 217G, 218G, 219G, 220G, 221G, 222G, 223G, 224G, 225G, 226G, 227G, 228G, 229G, 230G, 231G, 232G, 233G, 234G, 235G, 236G, 237G, 238G, 239G, 240G, 241G, 242G, 243G, 244G, 245G, 246G; and 247G-1047G;

1M, 2M, 3M, 4M, 5M, 6M, 7M, 8M, 9M, 10M, 11M, 12M, 13M, 14M, 15M, 16M, 17M, 18M, 19M, 20M, 21M, 22M, 23M, 24M, 25M, 26M, 27M, 28M, 29M, 30M, 31M, 32M, 33M, 34M, 35M, 36M, 37M, 38M, 39M, 40M, 41M, 42M, 43M, 44M, 45M, 46M, 47M, 48M, 49M, 50M, 51M, 52M, 53M, 54M, 55M, 56M, 57M, 58M, 59M, 60M, 61M, 62M, 63M, 64M, 65M, 66M, 67M, 68M, 69M, 70M, 71M, 72M, 73M, 74M, 75M, 76M, 77M, 78M, 79M, 80M, 81M, 82M, 83M, 84M, 85M, 86M, 87M, 88M, 89M, 90M, 91M, 92M, 93M, 94M, 95M, 96M, 97M, 98M, 99M, 100M, 101M, 102M, 103M, 104M, 105M, 106M, 107M, 108M, 109M, 110M, 111M, 112M, 113M, 114M, 115M, 116M, 117M, 118M, 119M, 120M, 121M, 122M, 123M, 124M, 125M, 126M, 127M, 128M, 129M, 130M, 131M, 132M, 133M, 134M, 135M, 136M, 137M, 138M, 139M, 140M, 141M, 142M, 143M, 144M, 145M, 146M, 147M, 148M, 149M, 150M, 151M, 152M, 153M, 154M, 155M, 156M, 157M, 158M, 159M, 160M, 161M, 162M, 163M, 164M, 165M, 166M, 167M, 168M, 169M, 170M, 171M, 172M, 173M, 174M, 175M, 176M, 177M, 178M, 179M, 180M, 181M, 182M, 183M, 184M, 185M, 186M, 187M, 188M, 189M, 190M, 191M, 192M, 193M, 194M, 195M, 196M, 197M, 198M, 199M, 200M, 201M, 202M, 203M, 204M, 205M, 206M, 207M, 208M, 209M, 210M, 211M, 212M, 213M, 214M, 215M, 216M, 217M, 218M, 219M, 220M, 221M, 222M, 223M, 224M, 225M, 226M, 227M, 228M, 229M, 230M, 231M, 232M, 233M, 234M, 235M, 236M, 237M, 238M, 239M, 240M, 241M, 242M, 243M, 244M, 245M, 246M; and 247M-1047M;

1J, 2J, 3J, 4J, 5J, 6J, 7.1, 8J, 9J, 10J, 11J, 12J, 13J, 14J, 15J, 16J, 17J, 18J, 19J, 20J, 21J, 22J, 23J, 24.1, 25J, 26J, 27J, 28J, 29J, 30J, 31J, 32J, 33J, 34J, 35J, 36J, 37J, 38J, 39J, 40J, 41J, 42J, 43J, 44J, 45J, 46J, 47J, 48.1, 49J, 50J, 51J, 52J, 53J, 54J, 55J, 56J, 57J, 58J, 59J, 60J, 61J, 62J, 63J, 64J, 65J, 66J, 67J, 68J, 69J, 70J, 71.1, 72J, 73.1, 74J, 75J, 76J, 77J, 78J, 79J, 80J, 81J, 82J, 83J, 84J, 85J, 86J, 87J, 88J, 89J, 90J, 91J, 92J, 93J, 94J, 95J, 96J, 97J, 98J, 99J, 100J, 101J, 102J, 103J, 104J, 105J, 106J, 107J, 108J, 109J, 110J, 111J, 112J, 113J, 114J, 115J, 116J, 117J, 118J, 119J, 120J, 121J, 122J, 123J, 124.1, 125J, 126J, 127J, 128J, 129J, 130J, 131J, 132J, 133J, 134J, 135J, 136J, 137J, 138J, 139J, 140J, 141J, 142J, 143J, 144J, 145.1, 146J, 147J, 148J, 149J, 150J, 151J, 152J, 153J, 154J, 155J, 156J, 157J, 158J, 159J, 160J, 161J, 162J, 163J, 164J, 165J, 166J, 167J, 168J, 169J, 170J, 171J, 172J, 173J, 174J, 175J, 176J, 177J, 178J, 179J, 180J, 181J, 182J, 183.1, 184J, 185J, 186.1, 187J, 188J, 189.1, 190J, 191J, 192J, 193J, 194J, 195J, 196J, 197J, 198J, 199J, 200J, 201J, 202J, 203J, 204J, 205J, 206J, 207J, 208J, 209J, 210J, 211J, 212J, 213J, 214J, 215J, 216J, 217J, 218J, 219J, 220J, 221J, 222J, 223J, 224J, 225J, 226J, 227J, 228J, 229J, 230J, 231J, 232J, 233J, 234.1, 235J, 236J, 237J, 238J, 239J, 240J, 241J, 242J, 243J, 244J, 245J, 246J; and 247J-1047J;

1L, 2L, 3L, 4L, 5L, 6L, 7L, 8L, 9L, 10L, 11L, 12L, 13L, 14L, 15L, 16L, 17L, 18L, 19L, 20L, 21L, 22L, 23L, 24L, 25L, 26L, 27L, 28L, 29L, 30L, 31L, 32L, 33L, 34L, 35L, 36L, 37L, 38L, 39L, 40L, 41L, 42L, 43L, 44L, 45L, 46L, 47L, 48L, 49L, 50L, 51L, 52L, 53L, 54L, 55L, 56L, 57L, 58L, 59L, 60L, 61L, 62L, 63L, 64L, 65L, 66L, 67L, 68L, 69L, 70L, 71L, 72L, 73L, 74L, 75L, 76L, 77L, 78L, 79L, 80L, 81L, 82L, 83L, 84L, 85L, 86L, 87L, 88L, 89L, 90L, 91L, 92L, 93L, 94L, 95L, 96L, 97L, 98L, 99L, 100L, 101L, 102L, 103L, 104L, 105L, 106L, 107L, 108L, 109L, 110L, 111L, 112L, 113L, 114L, 115L, 116L, 117L, 118L, 119L, 120L, 121L, 122L, 123L, 124L, 125L, 126L, 127L, 128L, 129L, 130L, 131L, 132L, 133L, 134L, 135L, 136L, 137L, 138L, 139L, 140L, 141L, 142L, 143L, 144L, 145L, 146L, 147L, 148L, 149L, 150L, 151L, 152L, 153L, 154L, 155L, 156L, 157L, 158L, 159L, 160L, 161L, 162L, 163L, 164L, 165L, 166L, 167L, 168L, 169L, 170L, 171L, 172L, 173L, 174L, 175L, 176L, 177L, 178L, 179L, 180L, 181L, 182L, 183L, 184L, 185L, 186L, 187L, 188L, 189L, 190L, 191L, 192L, 193L, 194L, 195L, 196L, 197L, 198L, 199L, 200L, 201L, 202L, 203L, 204L, 205L, 206L, 207L, 208L, 209L, 210L, 211L, 212L, 213L, 214L, 215L, 216L, 217L, 218L, 219L, 220L, 221L, 222L, 223L, 224L, 225L, 226L, 227L, 228L, 229L, 230L, 231L, 232L, 233L, 234L, 235L, 236L, 237L, 238L, 239L, 240L, 241L, 242L, 243L, 244L, 245L, 246L; and 247L-1047L;

1V, 2V, 3V, 4V, 5V, 6V, 7V, 8V, 9V, 10V, 11V, 12V, 13V, 14V, 15V, 16V, 17V, 18V, 19V, 20V, 21V, 22V, 23V, 24V, 25V, 26V, 27V, 28V, 29V, 30V, 31V, 32V, 33V, 34V, 35V, 36V, 37V, 38V, 39V, 40V, 41V, 42V, 43V, 44V, 45V, 46V, 47V, 48V, 49V, 50V, 51V, 52V, 53V, 54V, 55V, 56V, 57V, 58V, 59V, 60V, 61V, 62V, 63V, 64V, 65V, 66V, 67V, 68V, 69V, 70V, 71V, 72V, 73V, 74V, 75V, 76V, 77V, 78V, 79V, 80V, 81V, 82V, 83V, 84V, 85V, 86V, 87V, 88V, 89V, 90V, 91V, 92V, 93V, 94V, 95V, 96V, 97V, 98V, 99V, 100V, 101V, 102V, 103V, 104V, 105V, 106V, 107V, 108V, 109V, 110V, 111V, 112V, 113V, 114V, 115V, 116V, 117V, 118V, 119V, 120V, 121V, 122V, 123V, 124V, 125V, 126V, 127V, 128V, 129V, 130V, 131V, 132V, 133V, 134V, 135V, 136V, 137V, 138V, 139V, 140V, 141V, 142V, 143V, 144V, 145V, 146V, 147V, 148V, 149V, 150V, 151V, 152V, 153V, 154V, 155V, 156V, 157V, 158V, 159V, 160V, 161V, 162V, 163V, 164V, 165V, 166V, 167V, 168V, 169V, 170V, 171V, 172V, 173V, 174V, 175V, 176V, 177V, 178V, 179V, 180V, 181V, 182V, 183V, 184V, 185V, 186V, 187V, 188V, 189V, 190V, 191V, 192V, 193V, 194V, 195V, 196V, 197V, 198V, 199V, 200V, 201V, 202V, 203V, 204V, 205V, 206V, 207V, 208V, 209V, 210V, 211V, 212V, 213V, 214V, 215V, 216V, 217V, 218V, 219V, 220V, 221V, 222V, 223V, 224V, 225V, 226V, 227V, 228V, 229V, 230V, 231V, 232V, 233V, 234V, 235V, 236V, 237V, 238V, 239V, 240V, 241V, 242V, 243V, 244V, 245V, 246V; and 247V-1047V;

1KK, 2KK, 3KK, 4KK, 5KK, 6KK, 7KK, 8KK, 9KK, 10KK, 11KK, 12KK, 13KK, 14KK, 15KK, 16KK, 17KK, 18KK, 19KK, 20KK, 21KK, 22KK, 23KK, 24KK, 25KK, 26KK, 27KK, 28KK, 29KK, 30KK, 31KK, 32KK, 33KK, 34KK, 35KK, 36KK, 37KK, 38KK, 39KK, 40KK, 41KK, 42KK, 43KK, 44KK, 45KK, 46KK, 47KK, 48KK, 49KK, 50KK, 51KK, 52KK, 53KK, 54KK, 55KK, 56KK, 57KK, 58KK, 59KK, 60KK, 61KK, 62KK, 63KK, 64KK, 65KK, 66KK, 67KK, 68KK, 69KK, 70KK, 71KK, 72KK, 73KK, 74KK, 75KK, 76KK, 77KK, 78KK, 79KK, 80KK, 81KK, 82KK, 83KK, 84KK, 85KK, 86KK, 87KK, 88KK, 89KK, 90KK, 91KK, 92KK, 93KK, 94KK, 95KK, 96KK, 97KK, 98KK, 99KK, 100KK, 101KK, 102KK, 103KK, 104KK, 105KK, 106KK, 107KK, 108KK, 109KK, 110KK, 111KK, 112KK, 113KK, 114KK, 115KK, 116KK, 117KK, 118KK, 119KK, 120KK, 121KK, 122KK, 123KK, 124KK, 125KK, 126KK, 127KK, 128KK, 129KK, 130KK, 131KK, 132KK, 133KK, 134KK, 135KK, 136KK, 137KK, 138KK, 139KK, 140KK, 141KK, 142KK, 143KK, 144KK, 145KK, 146KK, 147KK, 148KK, 149KK, 150KK, 151KK, 152KK, 153KK, 154KK, 155KK, 156KK, 157KK, 158KK, 159KK, 160KK, 161KK, 162KK, 163KK, 164KK, 165KK, 166KK, 167KK, 168KK, 169KK, 170KK, 171KK, 172KK, 173KK, 174KK, 175KK, 176KK, 177KK, 178KK, 179KK, 180KK, 181KK, 182KK, 183KK, 184KK, 185KK, 186KK, 187KK, 188KK, 189KK, 190KK, 191KK, 192KK, 193KK, 194KK, 195KK, 196KK, 197KK, 198KK, 199KK, 200KK, 201KK, 202KK, 203KK, 204KK, 205KK, 206KK, 207KK, 208KK, 209KK, 210KK, 211KK, 212KK, 213KK, 214KK, 215KK, 216KK, 217KK, 218KK, 219KK, 220KK, 221KK, 222KK, 223KK, 224KK, 225KK, 226KK, 227KK, 228KK, 229KK, 230KK, 231KK, 232KK, 233KK, 234KK, 235KK, 236KK, 237KK, 238KK, 239KK, 240KK, 241KK, 242KK, 243KK, 244KK, 245KK, 246KK; and 247KK-1047KK;

1LL, 2LL, 3LL, 4LL, 5LL, 6LL, 7LL, 8LL, 9LL, 10LL, 11LL, 12LL, 13LL, 14LL, 15LL, 16LL, 17LL, 18LL, 19LL, 20LL, 21LL, 22LL, 23LL, 24LL, 25LL, 26LL, 27LL, 28LL, 29LL, 30LL, 31LL, 32LL, 33LL, 34LL, 35LL, 36LL, 37LL, 38LL, 39LL, 40LL, 41LL, 42LL, 43LL, 44LL, 45LL, 46LL, 47LL, 48LL, 49LL, 50LL, 51LL, 52LL, 53LL, 54LL, 55LL, 56LL, 57LL, 58LL, 59LL, 60LL, 61LL, 62LL, 63LL, 64LL, 65LL, 66LL, 67LL, 68LL, 69LL, TOLL, 71LL, 72LL, 73LL, 74LL, 75LL, 76LL, 77LL, 78LL, 79LL, 80LL, 81LL, 82LL, 83LL, 84LL, 85LL, 86LL, 87LL, 88LL, 89LL, 90LL, 91LL, 92LL, 93LL, 94LL, 95LL, 96LL, 97LL, 98LL, 99LL, 100LL, 101LL, 102LL, 103LL, 104LL, 105LL, 106LL, 107LL, 108LL, 109LL, 110LL, 111LL, 112LL, 113LL, 114LL, 115LL, 116LL, 117LL, 118LL, 119LL, 120LL, 121LL, 122LL, 123LL, 124LL, 125LL, 126LL, 127LL, 128LL, 129LL, 130LL, 131LL, 132LL, 133LL, 134LL, 135LL, 136LL, 137LL, 138LL, 139LL, 140LL, 141LL, 142LL, 143LL, 144LL, 145LL, 146LL, 147LL, 148LL, 149LL, 150LL, 151LL, 152LL, 153LL, 154LL, 155LL, 156LL, 157LL, 158LL, 159LL, 160LL, 161LL, 162LL, 163LL, 164LL, 165LL, 166LL, 167LL, 168LL, 169LL, 170LL, 171LL, 172LL, 173LL, 174LL, 175LL, 176LL, 177LL, 178LL, 179LL, 180LL, 181LL, 182LL, 183LL, 184LL, 185LL, 186LL, 187LL, 188LL, 189LL, 190LL, 191LL, 192LL, 193LL, 194LL, 195LL, 196LL, 197LL, 198LL, 199LL, 200LL, 201LL, 202LL, 203LL, 204LL, 205LL, 206LL, 207LL, 208LL, 209LL, 210LL, 211LL, 212LL, 213LL, 214LL, 215LL, 216LL, 217LL, 218LL, 219LL, 220LL, 221LL, 222LL, 223LL, 224LL, 225LL, 226LL, 227LL, 228LL, 229LL, 230LL, 231LL, 232LL, 233LL, 234LL, 235LL, 236LL, 237LL, 238LL, 239LL, 240LL, 241LL, 242LL, 243LL, 244LL, 245LL, 246LL; and 247LL-1047LL;

1XX, 2XX, 3XX, 4XX, 5XX, 6XX, 7XX, 8XX, 9XX, 10XX, 11XX, 12XX, 13XX, 14XX, 15XX, 16XX, 17XX, 18XX, 19XX, 20XX, 21XX, 22XX, 23XX, 24XX, 25XX, 26XX, 27XX, 28XX, 29XX, 30XX, 31XX, 32XX, 33XX, 34XX, 35XX, 36XX, 37XX, 38XX, 39XX, 40XX, 41XX, 42XX, 43XX, 44XX, 45XX, 46XX, 47XX, 48XX, 49XX, 50XX, 51XX, 52XX, 53XX, 54XX, 55XX, 56XX, 57XX, 58XX, 59XX, 60XX, 61XX, 62XX, 63XX, 64XX, 65XX, 66XX, 67XX, 68XX, 69XX, 70XX, 71XX, 72XX, 73XX, 74XX, 75XX, 76XX, 77XX, 78XX, 79XX, 80XX, 81XX, 82XX, 83XX, 84XX, 85XX, 86XX, 87XX, 88XX, 89XX, 90XX, 91XX, 92XX, 93XX, 94XX, 95XX, 96XX, 97XX, 98XX, 99XX, 100XX, 101XX, 102XX, 103XX, 104XX, 105XX, 106XX, 107XX, 108XX, 109XX, 110XX, 111XX, 112XX, 113XX, 114XX, 115XX, 116XX, 117XX, 118XX, 119XX, 120XX, 121XX, 122XX, 123XX, 124XX, 125XX, 126XX, 127XX, 128XX, 129XX, 130XX, 131XX, 132XX, 133XX, 134XX, 135XX, 136XX, 137XX, 138XX, 139XX, 140XX, 141XX, 142XX, 143XX, 144XX, 145XX, 146XX, 147XX, 148XX, 149XX, 150XX, 151XX, 152XX, 153XX, 154XX, 155XX, 156XX, 157XX, 158XX, 159XX, 160XX, 161XX, 162XX, 163XX, 164XX, 165XX, 166XX, 167XX, 168XX, 169XX, 170XX, 171XX, 172XX, 173XX, 174XX, 175XX, 176XX, 177XX, 178XX, 179XX, 180XX, 181XX, 182XX, 183XX, 184XX, 185XX, 186XX, 187XX, 188XX, 189XX, 190XX, 191XX, 192XX, 193XX, 194XX, 195XX, 196XX, 197XX, 198XX, 199XX, 200XX, 201XX, 202XX, 203XX, 204XX, 205XX, 206XX, 207XX, 208XX, 209XX, 210XX, 211XX, 212XX, 213XX, 214XX, 215XX, 216XX, 217XX, 218XX, 219XX, 220XX, 221XX, 222XX, 223XX, 224XX, 225XX, 226XX, 227XX, 228XX, 229XX, 230XX, 231XX, 232XX, 233XX, 234XX, 235XX, 236XX, 237XX, 238XX, 239XX, 240XX, 241XX, 242XX, 243XX, 244XX, 245XX, 246XX; and 247XX-1047XX;

In another composition aspect, the present invention is directed to fibrils comprising four or more of peptides comprising one or more of hexapeptides that generate a free energy of formation of −23 kcal/mol or less (i.e., more negative) when the hexapeptide sequence in subjected to the Rosetta-Profile algorithm.

In yet another compositions aspect, the present invention is directed to complexes comprising one or more fibrils of the present invention and one or more plasma proteins, including plasma proteins A-WW listed above.

Where the peptides of the present invention are formulated to be administered to a mammal, the formulation is typically chosen to provide a homogeneous solution comprising peptides. Peptides with amino acid subunits that are anionic at pH 7.4 are oftentimes formulated in water at a pH of 3-5. Peptides with amino acid subunits that are cationic at pH 7.4 are oftentimes formulated in water at pH 7.4 or a pH above 9 (e.g., 10). In certain cases, a solubility enhancer may be included in the formulation. Nonlimiting examples include DMSO and ethanol.

Compositions of the present invention are used to treat mammalian diseases. Compositions of the present invention may be used to treat inflammatory conditions. Nonlimiting examples of such inflammatory conditions include: multiple sclerosis; stroke; cardiac ischemia-reperfusion injury; retinal ischemia-reperfusion injury; macular degeneration; glaucoma; retinitis pigmentosa; rheumatoid arthritis; Type 1 diabetes; Type 2 diabetes; amyloid neuropathy; and amyloid nephropathy.

In a method aspect of the present invention, a method of treating an inflammatory condition is provided. The method comprises administration of a therapeutically effective amount of a composition of the present invention, in a suitable formulation, to a mammal experiencing an inflammatory condition.

Administration of the formulation may be performed by any method producing the desired therapeutic results. Nonlimiting examples of administration methods include: oral; sublingual; buccal; intranasal; intrapulmonary; intraceregral; intravenous; intramuscular; rectal; and, epidural.

Examples
Methods

EAE was induced by procedures previously described. See, Steinman, L. & Zamvil, S. S. How to successfully apply animal studies in experimental allergic encephalomyelitis to research on multiple sclerosis. Ann Neurol 60, 12-21, doi:10.1002/ana.20913 (2006). When animals exhibited hindlimb weakness they were injected in the peritoneum with either 1 μg of peptide or PBS daily. Sera from EAE mice were collected following two days of treatment with 10 μg HspB1 protein, 1 μg tau 623-628 (VQIVYK)(SEQ ID NO: 123), or PBS. Twenty-six cytokines were analyzed by multiplex-bead-analysis.

The capacity of the proteins and peptides to inhibit DTT induced aggregation of the beta chain of insulin was assayed using procedures described previously by several authors. See, for example, Hong, D. P. & Fink, A. L. Independent heterologous fibrillation of insulin and its B-chain peptide. Biochemistry 44, 16701-16709, doi:10.1021/bi051658y (2005). Also see, Hong, D. P., Ahmad, A. & Fink, A. L. Fibrillation of human insulin A and B chains. Biochemistry 45, 9342-9353, doi:10.1021/bi0604936 (2006). The aggregation was measured by the increase in absorption at 360 nm as a function of time over twenty minutes.

The relative amount of amyloid present in each solution was measured by combining solutions of the hexamers (200 mg 50 ml) dissolved in 100 mM MES pH 7.4 and 50 ml of a 1M buffer solution at the appropriate pH (glycine, pH 3; acetate, pH 4; citrate, pH 5; MES, pH 6; Tris, pH 7; Tris, pH 8; glycine, pH 9; bicarbonate, pH 10) and incubated at 37° C. ThT (10 ml of 10 mM solution) was added and the resultant emission fluorescence at 485 nm for each sample after excitation at 440 nm was measured using a SpectraMax 190 fluorescent microtiter plate reader.

The ligands for the amyloid fibrils were identified by incubating 50 mgs of biotinylated fibrils of Tau 623-628 with fresh human plasma at 37° C. for one hour after which 50 ml of streptavidin sepharose beads were added and the mixture was incubated an additional hour at 37° C. The resin was separated from the plasma, washed, and the ligands were eluted. The resin was separated from the eluted proteins as previously described. See, Rothbard, J. B. et al. Therapeutic effects of systemic administration of chaperone alphaB-crystallin associated with binding proinflammatory plasma proteins. J Biol Chem 287, 9708-9721, doi:M111.337691 [pii] 10.1074/jbc.M111.337691 (2012). The ligands were precipitated by the addition of TCA, and resultant precipitates were reconstituted, reduced and alkylated, trypsinized, and the resultant peptides were separated by HPLC. Liquid chromatography and mass spectral analysis were done as previously described. Id.

Experiments

1. Therapeutic Effectiveness of Amyloidogenic Hexamers.

Groups of ten mice were injected daily with 1 μg of the peptides listed in Table 1 (shown below) beginning at onset of hindlimb weakness. PBS or PBS containing 50% DMSO was injected in control littermates. Hexamers corresponding to residues 76-81, 89-94, and 623-628 of Tau effectively reduced the paralytic symptoms of EAE (Panel A of FIG. 1). Bars represent the duration of the treatment. Values in the graph represent the mean+/−S.E.M. *p<0.05 for SVNDLV (SEQ ID NO: 1059), LKVKVL (SEQ ID NO: 1060), VQIVYK (SEQ ID NO: 123) by the Mann Whitney U test. The shuffled analogs of HspB5 76-81 and Tau 623-628 were ineffective (Panels B&C) +p<0.05 for VQIVYK (SEQ ID NO: 123) by Mann Whitney U test. Hexamers corresponding to serum amyloid P 213-218, A beta A4 27-32, amylin 28-33 were therapeutic (Panel D of FIG. 1) *p<0.05 for GYVIIK (SEQ ID NO: 1050), NKGAII (SEQ ID NO: 1061), and SSTNVG (SEQ ID NO: 1053) by the Mann Whitney U test. Two poorly soluble peptides, prion 148-153 and A beta A4 35-40, were therapeutic when administered in 50% DMSO (Panel E) *p<0.05 for SNQNNF (SEQ ID NO: 1051) and ̂p<0.05 for MVGGVV (SEQ ID NO: 220) by the Mann Whitney U test. Administration of insulin B chain 11-16 did not statistically significantly reduce the symptoms of EAE (Panel F of FIG. 1).

TABLE 1

Hexameric peptides used in the study segregated by composition and/or

orientation in the fibril. Those whose crystal structure has been published

are in italics with their denoted classes, with the hydrophobic amino acids

highlighted in dark grey, acidic residues and

basic amino acids in light grey.

embedded image

2. Modulation of Plasma Cytokine Levels.

To determine whether a hexamer modulated the plasma cytokine levels equivalent to an intact small heat shock protein, groups of five mice were treated after the onset of hindlimb weakness with daily injections of either 1 μg of Tau 623-628 or 10 μg of HspB1 for two days and bled 12 hours later. Assaying the levels of 26 cytokines in the serum of untreated and treated mice revealed that administration of either HspB 1 or the tau hexamer resulted in a dramatic reduction of IL-6, with smaller reductions in IL-2, TGF-β, and IL-17. See FIG. 2.

3. pH Dependent Amyloid Formation.

The ability of each hexamer listed in Table 1 to form amyloid fibrils in aqueous buffers was examined. Each of the hexamers (200 mg 50 ml) was added to 100 ml of 100 mM MES pH 7.4 and combined with 50 ml of a 1M buffer solution at the appropriate pH (glycine, pH 3; acetate, pH 4; citrate, pH 5; MES, pH 6; Tris, pH 7; Tris, pH 8; glycine, pH 9; bicarbonate, pH 10) and incubated at 37° C. ThT (10 ml of 10 mM solution) was added and the resultant fluorescence at 485 nm from excitation at 440 nm was measured using a fluorescent plate reader. The peptides can be segregated into sets A-E: an anionic set forming fibrils at pH 3, pH 4, and pH 5 (A), a cationic group forming fibrils only at pH 10 (B), three cationic sequences that form amyloid fibrils at all pHs measured (C), a nonionizable set of sequences weakly binding ThT at all pHs (D), and a hydrophobic set of sequences that bind ThT at all pHs tested (E). Only a fraction of the peptides formed amyloid fibrils when dissolved in PBS pH 7.4, as measured by staining with thioflavin T (ThT). Their differential propensity to form amyloid fibrils was used to help segregate them into functional sets (Table 1).

The three peptides containing acidic residues, HspB5 76-81, insulin B chain 11-16, and insulin A chain 12-17, effectively formed amyloid fibrils only in a range of pH 3 to pH 5 when the acidic residue would be protonated (Panel A of FIG. 3). Reciprocally, a second set of two peptides containing lysine, HspB5 89-94 and amyloid b A4 27-32, bound ThT only at pH 10 when the lysine would not be protonated (Panel B of FIG. 3). In contrast, three peptides containing lysine, Tau 623-628, amyloid beta A4 16-21, and serum amyloid P 213-218, formed amyloid fibrils in buffers from pH 3-10, (Panel D of FIG. 3). Two other sets were identified, both of which contained nonionizable amino acids.

4. Correlation of ThT Staining with Molecular Chaperone Function.

To determine whether the ThT staining observed in Experiment 4 (above) correlated with molecular chaperone function, each of the peptides was assayed for its ability to inhibit insulin aggregation. The effect of ThT on insulin aggregation was measured as a control. Neither the addition of 10 mM ThT nor ThT bound to Tau 623-628 affected the rate of insulin aggregation in the former case or inhibition by the Tau peptide in the latter case. Insulin aggregation is pH dependent, only allowing a dynamic range for the assay at pH between 5 and 8, which allows only a fraction of the pH range used in the ThT staining experiments to be analyzed. See, Nielsen, L. et al. Effect of environmental factors on the kinetics of insulin fibril formation: elucidation of the molecular mechanism. Biochemistry 40, 6036-6046, doi:bi002555c [pii] (2001).

When HspB5 76-81 and Tau 623-628 were assayed for their ability to inhibit insulin aggregation, a clear distinction was apparent. The Tau peptide was a potent inhibitor at both pH 7.4 and pH 5, but the HspB5 peptide was only effective at pH 5 (Panels A and B of FIG. 4), which correlated with the pH dependence of ThT staining (Panels A and D of FIG. 3). The shuffled analogs of both peptides did not inhibit at either both pH 7.4 or pH 5. The inhibition with the Tau peptide was titratable, which allowed the calculation of an IC50 value for the Tau peptide of approximately 20 mg (Panel C of FIG. 4). The inhibition was dose dependent and both inhibitors delayed the appearance of the light scattering and consequently inhibited the nucleation step of the aggregation process (Panels A-C of FIG. 4).

The anionic peptide HspB5 76-81 is a molecular chaperone only at pH4 and not in more basic solutions, consistent with the observed ThT staining. Tau 623-628 inhibited insulin aggregation at pH 4 and pH 7.4. (Panel E of FIG. 4) Similarly, anionic peptides insulin alpha chain 11-16 and insulin beta chain 12-17 are molecular chaperones at pH 4, but not at pH 7.4.

5. Binding of Plasma Proteins by Amyloid Fibrils.

A comparison of the results from Experiments 1-4 showed that molecular chaperone function does not necessarily correlate with therapeutic activity; certain hexamers are effective therapeutics (see Experiment 1) but they do not exhibit molecular chaperone function. To determine whether the therapeutic compounds bind to specific plasma proteins, and accordingly can function as a therapeutic complex, Tau peptide was biotinylated and mixed with the unmodified hexamer to create a fibril that could be precipitated with streptavidin. Incubation of the two peptide mixtures with plasma from three separate MS patients, with subsequent precipitation, elution, tryptic cleavage, and mass spectral analyses, as described in the methods, resulted in the identification of 49 proteins whose relative concentrations were enhanced relative to the set of proteins identified when the streptavidin resin alone was used (see FIG. 5).

Forty-one of the proteins (84%) were among those identified by precipitation with the small heat shock protein, HspB5. See, Rothbard, J. B. et al. Therapeutic effects of systemic administration of chaperone alphaB-crystallin associated with binding proinflammatory plasma proteins. J Biol Chem 287, 9708-9721, doi:M111.337691 [pii] 10.1074/jbc.M111.337691 (2012). An extraordinarily high percentage of the proteins precipitated by the Tau fibril were members of the acute phase response (19/49, 39%), coagulation (13/49, 27%) or complement (11/49, 23%) pathways, which overall represented 33 of the 49 proteins (67%) precipitated. Strong evidence that the set of proteins constituted biologically relevant ligands was the presence of five proteins known to bind amyloid fibrils: Apolipoprotein A-I, A-IV, clusterin 20, apolipoproteins E 19, and transthyretin21. Many of the molecules described here also were identified in laser capture microdissected lesions from MS patients including Apolipoproteins AI, A2, B-100, and E, HspB5, tau, and amyloid precursor protein. See, Han, M. H. et al. Proteomic analysis of active multiple sclerosis lesions reveals therapeutic targets. Nature 451, 1076-1081 (2008). In addition, many of the molecules identified here also are known to modulate EAE. Earlier studies have shown that inhibition of angiotensin converting enzyme or angiotensin receptor, and inhibition of prothrombin all potently inhibit EAE. See Id. Also see, Platten, M. et al. Blocking angiotensin-converting enzyme induces potent regulatory T cells and modulates TH1- and TH17-mediated autoimmunity. Proc Natl Acad Sci USA 106, 14948-14953, doi:0903958106 [pii] 10.1073/pnas.0903958106 (2009). Interestingly, mice with genes encoding apolipoprotein E 23, tau, HspB5 1, and APP 24 individually knocked out all exhibit exacerbated EAE.

6. Therapeutic Effect of the Amyloid Fibrils does not Invoke Stereospecific Cell Surface Receptor

The comparative ability of Tau 623-628 (Ac VQIVYK CONH2)(SEQ ID NO: 123) and A beta 35-40 (Ac MVGGVV CONH2)(SEQ ID NO: 220) composed of L or D-amino acids to form amyloid fibrils was shown to be equivalent, within error, as defined by staining with Thioflavin T (FIG. 6a). The resultant stereoisomeric fibrils composed of the Tau peptide also inhibited insulin aggregation with very similar IC50 values (FIG. 6b). When used as therapeutic agents for EAE, equivalent reduction of paralytic symptoms were observed using peptides composed of D- or L-amino acids. This result has several important implications. The first is that the mechanism(s) will not involve binding to a stereospecific receptor. The second issue is that a common shared feature between the two stereoisomers is their molecular chaperone function. Not surprisingly, proteolytic cleavage of the peptides does not appear to be a major factor in limiting their mode of action, and lastly the set of possible relevant sequences has doubled with the shared activity of the peptides composed of D and L amino acids.

With respect the experimental results depicted in FIG. 6: A) L- and D-tau 623-628 and L- and D-Aβ 37-42 (200 μg/50 μl) were each added to 100 μl of 100 mM MES pH 7.4 and incubated at 37° C. ThT (10 μl of 10 μM solution) was added and the resultant fluorescence at 485 nm from excitation at 440 nm was measured using a fluorescent plate reader. The levels of ThT were similar in both sets of hexapeptides. B) L-tau and C) D-tau 623-628 inhibit insulin aggregation in a dose dependent manner with similar IC50 concentrations. D) Groups of ten mice were injected daily intraperitoneally for 10 days with 1 μg of L-tau or D-tau hexapeptides beginning at onset of hindlimb weakness. PBS was injected in littermates as control. Bar represents the duration of the treatment. Values in graph represent mean+/−S.E.M. *p<0.05 for L-tau and ̂p<0.05 for D-tau by Mann-Whitney U test.

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