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US 2011/0085989 A1Published application
Novel antimicrobial peptides
AssigneesThe Regents of the University of California, C3 Jian, Inc.
InventorsJian He, Randal H. Eckert, Fengxia Qi, Maxwell H. Anderson, Wenyuan Shi
Life of this patent
Estimating the term…
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.
Claims59
An antimicrobial peptide said 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;
- 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.
5 more elements
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.
4-7. (canceled)
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 and/or one or more beta amino acids.
17:
- (canceled)
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.
1 more element
20:
- The antimicrobial peptide of claim 19, wherein said peptide comprises an amino acid sequence selected from the group consisting of FKKFWKW FRRF (SEQ ID NO:31), 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.
1 more element
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 claim 1, wherein said peptide is labeled with a detectable label.
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 claim 1, 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.
11 more elements
30:
- The antimicrobial peptide of claim 27, wherein said second antimicrobacterial peptide is a peptide listed in Table 6.
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.
2 more elements
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.
2 more elements
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) (IC-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 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 His 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 claim 42 or 45, wherein said peptide comprises the amino acid sequence KLLK (SEQ ID NO:1) at one or both termini.
47:
- The antimicrobial peptide of claim 42 or 45, wherein said peptide comprises the amino acid sequence KLLK (SEQ ID NO:1) at one or both termini wherein said KLLK is joined to the terminus by a linker.
48:
- The antimicrobial peptide of claim 27, wherein said linker is GAT (SEQ ID NO:2).
49:
- The antimicrobial peptide of claim 42, wherein said peptide comprises the amino acid sequence FHFFHHFFHFFHHF (SEQ ID NO:3).
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), KFVKLI LAY (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 VWVHKHVLF (SEQ ID NO:124) (S3L8-2), WRWRARWRWRLRWRF (SEQ ID NO:125) (S3L8-3), WR1HLRARLHVKFRF (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).
55:
- A pharmaceutical formulation comprising an antimicrobial peptide of claim 1;
- and
- 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 home health care produce comprising:
- an antimicrobial peptide according to claim 1, wherein said antimicrobial peptide is contained in a product selected from the group consisting of toothpaste, mouthwash, a tooth whitening strip or solution, a 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 claim 1, in an amount sufficient to inhibit growth and/or proliferation of said bacterium.
60-66. (canceled)
67:
- A method of inhibiting the formation of dental caries, said method comprising contacting teeth and or oral mucosa with a peptide according to claim 1 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.
Dependencies are inferred from the claim text.