NOVEL ANTIMICROBIAL PEPTIDES

• US 20080286210A1
• Filed: 01/15/2008
• Published: 11/20/2008
• Est. Priority Date: 01/16/2007
• Status: Active Grant

• Current Assignee: C3 Jian, Inc., Regents of the University of California (University of California)
• Original Assignee: Regents of the University of California (University of California)
• Inventors: Shi, Wenyuan, Anderson, Maxwell H., He, Jian, Qi, Fengxia, Eckert, Randal H.
• Granted Patent: US 7,713,927 B2
• Time in Patent Office: Days
• US Class Current: 424/45
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 47/60 the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
• CPC Class Codes: A61K 8/64 Proteins; Peptides; Derivatives or degradation products thereof
• CPC Class Codes: A61K 9/0043 Nose
• CPC Class Codes: A61K 9/0056 Mouth soluble or dispersible forms; Suckable, eatable, chewable coherent forms; Forms rapidly disintegrating in the mouth; Lozenges; Lollipops; Bite capsules; Baked products; Baits or other oral forms for animals
• CPC Class Codes: A61K 9/006 Oral mucosa, e.g. mucoadhesive forms, sublingual droplets; Buccal patches or films; Buccal sprays
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: A61Q 11/00 Preparations for care of the teeth, of the oral cavity or of dentures; Dentifrices, e.g. toothpastes; Mouth rinses
• CPC Class Codes: A61Q 17/005 Antimicrobial preparations
• CPC Class Codes: C07K 14/001 by chemical synthesis
• CPC Class Codes: C07K 14/4723 Cationic antimicrobial peptides, e.g. defensins
• CPC Class Codes: C07K 7/06 having 5 to 11 amino acids
• CPC Class Codes: C07K 7/08 having 12 to 20 amino acids gastrins C07K14/595; somatostatins C07K14/655; melanotropins C07K14/68

Abstract

This invention provides novel antimicrobial peptides that are effective to inhibit growth and/or proliferation of various gram positive bacteria. In particular, the peptides are effective against Streptococcus mutans a common oral pathogen and the causative agent of dental caries.

Claims

1. An antimicrobial peptide said peptide comprising the following amino acid motif or a circular permutation of the following amino acid motif:

(H¹C¹C²H²H³C³H³H⁵C⁴C⁵)_n

wherein;

n ranges from 1 to 5 and can increment by units of 0.1;

H¹, H², H³, H⁴, and H⁵ are independently selected hydrophobic or hydrophilic amino acids;

C¹, C², C³, C⁴, and C⁵ are independently selected uncharged amino acids, positively charged amino acids, or negatively charged amino acids;

said peptide forms an alpha helix; and

said peptide is effective to kill or inhibit the growth or proliferation of Streptococcus mutans in culture.

2. The antimicrobial peptide of claim 1, wherein said peptide excludes the amino acid sequence FKKFWKW FRRF (SEQ ID NO:

31).

3. The antimicrobial peptide of claim 1, wherein C¹, C², C³, C⁴, and C⁵ are independently selected positively charged amino acids, or negatively charged amino acids.

4. The antimicrobial peptide of claim 1, wherein said uncharged amino acids are independently selected from the group consisting of S, T, and Y.

5. The antimicrobial peptide of claim 1, wherein said positively charged amino acids are independently selected from the group consisting of K, R, and H.

6. The antimicrobial peptide of claim 1, wherein said negatively charged amino acids are independently selected from the group consisting of N, Q, D, and E.

7. The antimicrobial peptide of claim 1, wherein said hydrophobic or hydrophilic amino acids are independently selected from the group consisting of using L, I, V, W, and F.

8. The antimicrobial peptide of claim 1, wherein:

said uncharged amino acids, positively charged amino acids, or negatively charged amino acids are independently selected from the group consisting of S, T, Y, K, R, H, N, Q, D, and E; and

said hydrophobic or hydrophilic amino acids are independently selected from the group consisting of using L, I, V, W, and F.

9. The antimicrobial peptide of claim 1, wherein said peptide further comprises a free amine at the carboxyl terminus.

10. The antimicrobial peptide of claim 9, wherein said peptide comprises a free amine at the carboxyl terminus that is provided by arginine or lysine.

11. The antimicrobial peptide of claim 9, wherein said peptide comprises a free amine at the carboxyl terminus that is provided by an amidated non-cationic residue.

12. The antimicrobial peptide of claim 1, wherein said peptide is pegylated.

13. The antimicrobial peptide of claim 1, wherein said peptide bears one or more protecting groups.

14. The antimicrobial peptide of claim 1, wherein said peptide bears one or more protecting groups selected from the group consisting of acetyl, amide, and 3 to 20 carbon alkyl groups, Fmoc, Tboc, 9-fluoreneacetyl group, 1-fluorenecarboxylic group, 9-florenecarboxylic group, 9-fluorenone-1-carboxylic group, benzyloxycarbonyl, Xanthyl (Xan), Trityl (Trt), 4-methyltrityl (Mtt), 4-methoxytrityl (Mmt), 4-methoxy-2,3,6-trimethyl-benzenesulphonyl (Mtr), Mesitylene-2-sulphonyl (Mts), 4,4-dimethoxybenzhydryl (Mbh), Tosyl (Tos), 2,2,5,7,8-pentamethyl chroman-6-sulphonyl (Pmc), 4-methylbenzyl (MeBzl), 4-methoxybenzyl (MeOBzl), Benzyloxy (BzlO), Benzyl (Bzl), Benzoyl (Bz), 3-nitro-2-pyridinesulphenyl (Npys), 1-(4,4-dimentyl-2,6-diaxocyclohexylidene)ethyl (Dde), 2,6-dichlorobenzyl (2,6-DiCl-Bzl), 2-chlorobenzyloxycarbonyl (2-Cl-Z), 2-bromobenzyloxycarbonyl (2-Br-Z), Benzyloxymethyl (Bom), t-butoxycarbonyl (Boc), cyclohexyloxy (cHxO), t-butoxymethyl (Bum), t-butoxy (tBuO), t-Butyl (tBu), Acetyl (Ac), and Trifluoroacetyl (TFA).

15. The antimicrobial peptide of claim 1, wherein said peptide comprises all naturally occurring amino acids.

16. The antimicrobial peptide of claim 1, wherein said peptide comprises one or more “

D”

amino acids.

17. The antimicrobial peptide of claim 1, wherein said peptide comprises one or more beta amino acids.

18. The antimicrobial peptide of claim 1, wherein said peptide has at least a +3 net positive charge at physiological pH.

19. The antimicrobial peptide of claim 1, wherein:

n is 1.1; and

said peptide comprises the motif H¹C¹C²H²H³C³H⁴H⁵C⁴C⁵H⁶, where H⁶ is an independently selected hydrophobic amino acid.

20. The antimicrobial peptide of claim 19, wherein said peptide comprises an amino acid sequence selected from the group consisting of FKKFWKWFRRF (SEQ ID NO:

31) (B-33), LKRFLKWFKRF (SEQ ID NO;

32) (B-34), KLFKRWKHLFR (SEQ ID NO;

33) (B-35), RLLKRFKHLFK (SEQ ID NO;

34) (B-36), FKTFLKWLHRF (SEQ ID NO;

35) (B-37), IKQLLHFFQRF (SEQ ID NO;

36) (B-38), KLLQTFKQIFR (SEQ ID NO;

37) (B-39), RILKELKNLFK (SEQ ID NO;

38) (B-40), LKQFVHFIHRF (SEQ ID NO;

39) (B-41), VKTLLHIFQRF (SEQ ID NO;

40) (B-42), KLVEQLKEIFR (SEQ ID NO;

41) (B-43), RVLQEIKQILK (SEQ ID NO;

42) (B-44), VKNLAELVHRF (SEQ ID NO;

43) (B-45), ATHLLHALQRF (SEQ ID NO;

44) (B-46), KLAENVKEILR (SEQ ID NO;

45) (B-47), RALHEAKEALK (SEQ ID NO;

46) (B-48), FHYFWHWFHRF (SEQ ID NO;

47) (B-49), LYHFLHWFQRF (SEQ ID NO;

48) (B-50), YLFQTWQHLFR (SEQ ID NO;

49) (B-51), YLLTEFQHLFK (SEQ ID NO;

50) (B-52), FKTFLQWLHRF (SEQ ID NO;

51) (B-53), IKTLLHFFQRF (SEQ ID NO;

52) (B-54), KLLQTFNQIFR (SEQ ID NO;

53) (B-55), TILQSLKNIFK (SEQ ID NO;

54) (B-56), LKQFVKFIHRF (SEQ ID NO;

55) (B-57), VKQLLKIFNRF (SEQ ID NO;

56) (B-58), KLVQQLKNIFR (SEQ ID NO;

57) (B-59), RVLNQVKQILK (SEQ ID NO;

58) (B-60), VKKLAKLVRRF (SEQ ID NO;

59) (B-61), AKRLLKVLKRF (SEQ ID NO;

60) (B-62), KLAQKVKRVLR (SEQ ID NO;

61) (B-63), and RALKRIKHVLK (SEQ ID NO;

62) (B-64).

21. The antimicrobial peptide of claim 20, wherein said peptide is amidated.

22. The antimicrobial peptide of claim 1, wherein:

n is 1.4; and

said peptide comprises the motif H¹C¹C²H²H³C³H⁴H⁵C⁴C⁵H⁶C⁶C⁷H⁷, where H⁶, and H⁷ are independently selected hydrophobic amino acids, and C⁶ and C⁷ are independently selected positively or negatively charged amino acids.

23. The antimicrobial peptide of claim 22, wherein said peptide comprises an amino acid sequence selected from the group consisting of KLKKLLKKLKKLLK (SEQ ID NO:

64) (α

-5), LKLLKKLLKLLKKF (SEQ ID NO;

65) (α

-6), LQLLKQLLKLLKQF (SEQ ID NO;

66) (α

-7), RWRRWWRHFHHFFH (SEQ ID NO;

68) (α

-9), KLKKLLKRWRRWWR (SEQ ID NO;

69) (α

-10), RWRRLLKKLHHLLH (SEQ ID NO;

70) (α

-11), and KLKKLLKHLHHLLH (SEQ ID NO;

71) (α

-12).

24. The antimicrobial peptide of claim 23, wherein said peptide is amidated.

25. The antimicrobial peptide of any one of claims 1, 20, or 23, wherein said peptide is labeled with a detectable label.

26. The antimicrobial peptide of claim 25, wherein said peptide is labeled with a detectable label selected from the group consisting of an enzymatic label, a fluorescent label, a colorimetric label, a spin label, and a radioactive label.

27. The antimicrobial peptide of any one of claims 1, 20, 23, or 43 wherein said peptide is attached to a second antimicrobial peptide thereby forming a compound antimicrobial peptide.

28. The antimicrobial peptide of claim 27, wherein said compound antimicrobial peptide is a fusion protein.

29. The antimicrobial peptide of claim 27, wherein said second antimicrobial peptide is:

a peptide comprising the following amino acid motif or a circular permutation of the following amino acid motif;

(H¹C¹C²H²H³C³H⁴H⁵C⁴C⁵)_n

wherein;

n ranges from 1 to 5 and can increment by units of 0.1;

H¹, H², H³, H⁴, and H⁵ are independently selected hydrophobic or hydrophilic amino acids; and

C¹, C², C³, C⁴, and C⁵ are independently selected neutral amino acids, positively charged amino acids, or negatively charged amino acids;

or

said peptide comprising the seven contiguous amino acids wherein;

all but two amino acids are Arg or Trp, or derivatives or analogues thereof;

the two non-Arg or Trp amino acids are Lys or Phe, or derivatives or analogues thereof; and

the N-terminal residue is Arg or a derivative or analogue thereof.

30. The antimicrobial peptide of claim 27, wherein said second antimicrobacterial peptide is a peptide listed in Table 6.

48. The antimicrobial peptide of claim 27, wherein said linker is GAT (SEQ ID NO:

167).

55. A pharmaceutical formulation comprising an antimicrobial peptide of any one of claims 1, 31, or 42;

and

a pharmaceutically acceptable excipient.

56. The pharmaceutical formulation of claim 55, wherein said formulation is a unit dosage formulation.

57. The pharmaceutical formulation of claim 55, wherein said excipient is acceptable for administration to an oral mucosa.

58. A health care product comprising

an antimicrobial peptide according to any of claims 1, 31, or 42, wherein said antimicrobial peptide is contained in a product selected from the group consisting of toothpaste, mouthwash, a tooth whitening strip or solution, s contact lens storage, wetting, or cleaning solution, dental floss, a toothpick, a toothbrush bristle, an oral spray, an oral lozenge, a nasal spray, an aerosolizer for oral and/or nasal application, and a wound dressing.

59. A method of inhibiting the growth and/or proliferation of a bacterium, said method comprising contacting said bacterium with a peptide according to any one of claims 1, 31, or 42, in an amount sufficient to inhibit growth and/or proliferation of said bacterium.

60. The method of claim 59, wherein said amount is an amount sufficient to kill said bacterium.

61. The method of claim 59, wherein said bacterium is a Gram positive bacterium.

62. The method of claim 59, wherein said bacterium is a Gram positive oral bacterium.

63. The method of claim 61, wherein said bacterium is a Streptococcus sp.

64. The method of claim 61, wherein said bacterium is a Streptococcus mutans.

65. The method of claim 59, wherein said contacting comprises contacting a mucosal surface.

66. The method of claim 59, wherein said contacting comprises contacting an oral mucosa.

67. A method of inhibiting the formation of dental caries, said method comprising contacting teeth and or oral mucosa with a peptide according to any one of claims 1, 31, or 42 in an amount sufficient to inhibit growth and/or proliferation of S. mutans.

68. The method of claim 59, wherein contacting comprises contacting said teeth and/or oral mucosa with a composition selected from the group consisting of a toothpaste, a mouthwash, a whitening strip or solution, a lozenge, an aerosol, and a swab.

31. An antimicrobial peptide said peptide comprising seven contiguous amino acids wherein:

all but two amino acids are Arg or Trp, or derivatives or analogues thereof;

the two non-Arg or Trp amino acids are Lys or Phe, or derivatives or analogues thereof; and

the N-terminal residue is Arg or a derivative or analogue thereof.

32. The antimicrobial peptide of claim 31, wherein:

all but two amino acids are Arg or Trp;

the two non-Arg or Trp amino acids are Lys or Phe; and

the N-terminal residue is Arg.

33. The antimicrobial peptide of claim 31, wherein said peptide comprises an amino acid sequence selected from the group consisting of RRRRWWW (SEQ ID NO:

72) (1C-1), RRWWRRW (SEQ ID NO;

73) (1C-2), RRRWWWR (SEQ ID NO;

74) (1C-3), RWRWRWR (SEQ ID NO;

75) (1C-4), RRRFWWR (SEQ ID NO;

76) (2C-1), RRWWRRF (SEQ ID NO;

77) (2C-2), RRRWWWF (SEQ ID NO;

78) (2C-3), RWRWRWF (SEQ ID NO;

79) (2C-4), RRRRWWK (SEQ ID NO;

80) (3C-1), RRWWRRK (SEQ ID NO;

81) (3C-2), RRRWWWK (SEQ ID NO;

82) (3C-3), RWRWRWK (SEQ ID NO;

83) (3C-4), RRRKWWK (SEQ ID NO;

84) (4C-1), RRWKRRK (SEQ ID NO;

85) (4C-2), RRRKWWK (SEQ ID NO;

86) (4C-3), and RWRKRWK (SEQ ID NO;

87) (4C-4).

34. The antimicrobial peptide of claim 31, wherein said peptide is pegylated.

35. The antimicrobial peptide of claim 31, wherein said peptide bears one or more protecting groups.

36. The antimicrobial peptide of claim 31, wherein said peptide bears one or more protecting groups selected from the group consisting of acetyl, amide, and 3 to 20 carbon alkyl groups, Fmoc, Tboc, 9-fluoreneacetyl group, 1-fluorenecarboxylic group, 9-florenecarboxylic group, 9-fluorenone-1-carboxylic group, benzyloxycarbonyl, Xanthyl (Xan), Trityl (Trt), 4-methyltrityl (Mtt), 4-methoxytrityl (Mmt), 4-methoxy-2,3,6-trimethyl-benzenesulphonyl (Mtr), Mesitylene-2-sulphonyl (Mts), 4,4-dimethoxybenzhydryl (Mbh), Tosyl (Tos), 2,2,5,7,8-pentamethyl chroman-6-sulphonyl (Pmc), 4-methylbenzyl (MeBzl), 4-methoxybenzyl (MeOBzl), Benzyloxy (BzlO), Benzyl (Bzl), Benzoyl (Bz), 3-nitro-2-pyridinesulphenyl (Npys), 1-(4,4-dimentyl-2,6-diaxocyclohexylidene)ethyl (Dde), 2,6-dichlorobenzyl (2,6-DiCl-Bzl), 2-chlorobenzyloxycarbonyl (2-Cl-Z), 2-bromobenzyloxycarbonyl (2-Br-Z), Benzyloxymethyl (Bom), t-butoxycarbonyl (Boc), cyclohexyloxy (cHxO), t-butoxymethyl (Bum), t-butoxy (tBuO), t-Butyl (tBu), Acetyl (Ac), and Trifluoroacetyl (TFA).

37. The antimicrobial peptide of claim 31, wherein said peptide comprises all naturally occurring amino acids.

38. The antimicrobial peptide of claim 31, wherein said peptide comprises one or more “

D”

amino acids.

39. The antimicrobial peptide of claim 31, wherein said peptide comprises one or more beta amino acids.

40. The antimicrobial peptide of claim 31, wherein said peptide is labeled with a detectable label.

41. The antimicrobial peptide of claim 40, wherein said peptide is labeled with a detectable label selected from the group consisting of an enzymatic label, a fluorescent label, a colorimetric label, a spin label, and a radioactive label.

42. An acid-activated antimicrobial peptide, said peptide comprising:

an amphipathic helical peptide ranging in length from about 7 to about 11 amino acids wherein the majority of charged residues are H is or a derivative or analogue thereof that caries a cationic charge at an acidic pH, wherein said peptide has substantially no antimicrobial activity at neutral pH, but has antimicrobial activity against S. mutans at an acidic pH.

43. The antimicrobial peptide of claim 42, wherein all charged residues are His or a derivative or analogue thereof that caries a cationic charge at an acidic pH.

44. The antimicrobial peptide of claim 42, wherein said acidic pH is pH 6 or lower.

45. The antimicrobial peptide of claim 42, wherein said peptide comprises alternating repeats of HH and FF.

46. The antimicrobial peptide of claims 42 or 45, wherein said peptide comprises the amino acid sequence KLLK (SEQ ID NO:

166) at one or both termini.

47. The antimicrobial peptide of claims 42 or 45, wherein said peptide comprises the amino acid sequence KLLK (SEQ ID NO:

166) at one or both termini wherein said KLLK (SEQ ID NO;

166) is joined to the terminus by a linker.

49. The antimicrobial peptide of claim 42, wherein said peptide comprises the amino acid sequence FHFFHHFFHFFHHF (SEQ ID NO:

110).

50. The antimicrobial peptide of claim 42, wherein said peptide comprises an amino acid sequence selected from the group consisting of HHFFHHFHHFFHHF (SEQ ID NO:

109) (AA-1), FHFFHHFFHFFHHF (SEQ ID NO;

110) (AA-2), KLLKGATFHFFHHFFHFFHHF (SEQ ID NO;

111) (AA-3), KLLKFHFFHHFFHFFHHF (SEQ ID NO;

112) (AA-4), FHFFHHFFHFFHHFKLLK (SEQ ID NO;

113) (AA-5), FHYFWHWFHRF (SEQ ID NO;

114) (AA-6), and LYHFLHWFQRF (SEQ ID NO;

115) (AA-7).

51. The antimicrobial peptide of claim 42, wherein said peptide bears one or more protecting groups.

52. The antimicrobial peptide of claim 42, wherein said peptide bears one or more protecting groups selected from the group consisting of acetyl, amide, and 3 to 20 carbon alkyl groups, Fmoc, Tboc, 9-fluoreneacetyl group, 1-fluorenecarboxylic group, 9-florenecarboxylic group, 9-fluorenone-1-carboxylic group, benzyloxycarbonyl, Xanthyl (Xan), Trityl (Trt), 4-methyltrityl (Mtt), 4-methoxytrityl (Mmt), 4-methoxy-2,3,6-trimethyl-benzenesulphonyl (Mtr), Mesitylene-2-sulphonyl (Mts), 4,4-dimethoxybenzhydryl (Mbh), Tosyl (Tos), 2,2,5,7,8-pentamethyl chroman-6-sulphonyl (Pmc), 4-methylbenzyl (MeBzl), 4-methoxybenzyl (MeOBzl), Benzyloxy (BzlO), Benzyl (Bzl), Benzoyl (Bz), 3-nitro-2-pyridinesulphenyl (Npys), 1-(4,4-dimentyl-2,6-diaxocyclohexylidene)ethyl (Dde), 2,6-dichlorobenzyl (2,6-DiCl-Bzl), 2-chlorobenzyloxycarbonyl (2-Cl-Z), 2-bromobenzyloxycarbonyl (2-Br-Z), Benzyloxymethyl (Bom), t-butoxycarbonyl (Boc), cyclohexyloxy (cHxO), t-butoxymethyl (Bum), t-butoxy (tBuO), t-Butyl (tBu), Acetyl (Ac), and Trifluoroacetyl (TFA).

53. An antimicrobial peptide comprising a peptide selected from the group consisting of LKQKLKILF (SEQ ID NO:

116 (S6L1-2), LKQLKAGIY (SEQ ID NO;

117) (S6L1-3), VGKCVKLLY (SEQ ID NO;

118) (S6L1-4), KFVKLILAY (SEQ ID NO;

119) (S6L1-5), KLVKLVFLY (SEQ ID NO;

120) (S6L1-6), IKVFAKQKY (SEQ ID NO;

121) (S6L1-7), and RFRHFQERY (SEQ ID NO;

122) (S6L1-8).

54. An antimicrobial peptide comprising a peptide selected from the group consisting of FVFRHKWVWKHRFLF (SEQ ID NO:

123) (S3L8-1), VFI VWVHKIIVLF (SEQ ID NO;

124) (S3L8-2), WRWRARWRWRLRWRF (SEQ ID NO;

125) (S3L8-3), WRlHLRARLHVKFRF (SEQ ID NO;

126) (S3L8-4), LRIHARFKVHIRLKF (SEQ ID NO;

127) (S3L8-5), FHIKFRVHLKVRFHF (SEQ ID NO;

128) (S3L8-6), FHVK1HFRLHVKFHF (SEQ ID NO;

129) (S3L8-7), LHIHAHFHVHIHLHF (SEQ ID NO;

130) (S3L8-8), FKIHFRLKVHIRFKF (SEQ ID NO;

131) (S3L8-9), FKAHIRFKLRVKFHF (SEQ ID NO;

132) (S3L8-10), LKAKIKFKVKLKIKF (SEQ ID NO;

133) (S3L8-11), WIWKHKFLHRHFLF (SEQ ID NO;

134) (S3L8-12), VFLHRHVIKHKLVF (SEQ ID NO;

135) (S3L8-13), FLHKHVLRHRIVF (SEQ ID NO;

136) (S3L8-14), VFKHKIVHRHILF (SEQ ID NO;

137) (S3L8-15), FLFKHLFLHRIFF (SEQ ID NO;

138) (S3L8-16), LFKHILIHRVIF (SEQ ID NO;

139) (S3L8-17), FLHKHLFKHKLF (SEQ ID NO;

140) (S3L8-18), VFRHRFIHRHVF (SEQ ID NO;

141) (S3L8-19), FIHKLVHKHVLF (SEQ ID NO;

142) (S3L8-20), VLRHLFRHRIVF (SEQ ID NO;

143) (S3L8-21), LVHKLILRHLLF (SEQ ID NO;

144) (S3L8-22), VFKRVLI HKLIF (SEQ ID NO;

145) (S3L8-23), IVRKFLFRHKVF (SEQ ID NO;

146) (S3L8-24), VLKHVIAHKRLF (SEQ ID NO;

147) (S3L8-25), FIRKFLFKHLF (SEQ ID NO;

148) (S3L8-26), VIRHVWVRKLF (SEQ ID NO;

149) (S3L8-27), FLFRHRFRHRLVF (SEQ ID NO;

150) (S3L8-28), LFLHKHAKHKFLF (SEQ ID NO;

151) (S3L8-29), FKHKFKHKFIF (SEQ ID NO;

152) (S3L8-30), LRHRLRHRLIF (SEQ ID NO;

153) (S3L8-31), LILKFLFKFVF (SEQ ID NO;

154) (S3L8-32), VLIRILVRVIF (SEQ ID NO;

155) (S3L8-33), FRHRFRHRF (SEQ ID NO;

156) (S3L8-34), LKHKLKHKF (SEQ ID NO;

157) (S3L8-35), FKFKHKLIF (SEQ ID NO;

158) (S3L8-36), LRLRHRVLF (SEQ ID NO;

159) (S3L8-37), FKFLFKFLF (SEQ ID NO;

160) (S3L8-38), LRLFLRWLF (SEQ ID NO;

161) (S3L8-39), FKFLFKHKF (SEQ ID NO;

162) (S3L8-40), LRLFLRHRF (SEQ ID NO;

163) (S3L8-41), FKFLFKF (SEQ ID NO;

164) (S3L8-42), and LRLFLRF (SEQ ID NO;

165) (S3L8-43).