METHODS FOR STIMULATING FIBROBLAST PROLIFERATION USING SUBSTANCE P ANALOGS

1 . A method of stimulating or promoting fibroblast cell proliferation comprising adding one or more substance P analogs to substantially purified fibroblast cells wherein said substance P analog is according to Formula (J):

Z 1 -Xaa 1 -Xaa 2 -Xaa 3 -Xaa 4 -Xaa 5 -Xaa 6 -Xaa 7 -Xaa 8 -Xaa 9 -

Xaa 10 -Xaa 11 -Z 2 (I)

or a pharmaceutically acceptable salt thereof, wherein:

Xaa 1 is Arg, Lys, 6-N methyllysine, or (6-N, 6-N) dimethyllysine;

Xaa 2 is Pro or Ala;

Xaa 3 is Lys, Arg, 6-N-methyllysine, or (6-N, 6-N) dimethyllysine;

Xaa 4 is Pro or Ala;

Xaa 5 is Gln or Asn;

Xaa 6 is Gln or Asn;

Xaa 7 is Tyr, Phe, or Phe substituted with chlorine at position 2, 3, or 4;

Xaa 8 is Tyr, Phe, or Phe substituted with chlorine at position 2, 3, or 4;

Xaa 9 is Gly, Pro, Ala, or N-methylglycine;

Xaa 10 is Leu, Val, Ile, norleucine, Met, Met sulfoxide, Met sulfone, N-methylleucine, or N-methylvaline;

Xaa 11 is Met, Met sulfoxide, Met sulfone, or norleucine;

Z 1 is R 2 N— or RC(O)NR—;

Z 2 is —C(O)NR 2 or —C(O)OR or a salt thereof,

each R is independently R is —H, (C 1 -C 6 ) alkyl, (C 1 -C 6 ) alkenyl, (C 1 -C 6 ) alkynyl, (C 5 -C 20 ) aryl, (C 6 -C 26 ) alkaryl, 5-20 membered heteroaryl, or 6-26 membered alkheteroaryl; and

each “-” between residues Xaa 1 through Xaa 11 independently designates an amide linkage, a substitute amide linkage, or an isostere of an amide, wherein the substance P analog is not substance P (SEQ ID NO.: 1).

2 . The method of claim 1 wherein

Xaa 1 is Arg;

Xaa 2 is Pro;

Xaa 3 is Lys;

Xaa 4 is Pro;

Xaa 5 is Gln;

Xaa 6 is Gln;

Xaa 7 is Tyr, Phe, or Phe substituted with chlorine at position 4;

Xaa 8 is Tyr, Phe, or Phe substituted with chlorine at position 4;

Xaa 9 is Gly, Pro, or N-methylglycine;

Xaa 10 is Leu; and

Xaa 11 is Met, Met sulfoxide, Met sulfone, or norleucine.

3 . The method of claim 1 wherein the “-” between residues Xaa 1 through Xaa 11 designates

—C(O)NH—;

Z 1 is H 2 N—; and Z 2 is —C(O)NH 2 .

4 . The method of claim 1 wherein the substance P analog is selected from the group consisting of:

RPKPQQFFGLNle;

(SEQ ID NO.:2)

RPKPQQFFPLM;

(SEQ ID NO.:3)

RPKPQQFFMeGlyLM;

(SEQ ID NO.:4)

RPKPQQFTGLM;

(SEQ ID NO.:5)

RPKPQQF(4-Cl)F(4-Cl)GLM;

(SEQ ID NO.:6)

RPKPQQFFGLM(O);

(SEQ ID NO.:7)

RPKPQQFFMeGlyLM(O);

(SEQ ID NO.:8)

RPKPQQFFMeGLM(O 2 );

(SEQ ID NO.:9)

and

RPKPQQFFMeGlyLM(O 2 ).

(SEQ ID NO.:10)

5 . The method of claim 1 wherein the substance P analog is

Z 1 -RPKPQQFFMeGlyLM(O 2 )-Z 2 ;

wherein

Z 1 is NH 2 and Z 2 is C(O)NH 2 .

6 . The method of claim 1 wherein the fibroblast cells are human fibroblast cells.

7 . The method of claim 1 wherein the fibroblast cells are dermal fibroblast cells.

8 . The method of claim 1 , further comprising administering said fibroblast cells to a subject to treat a defect of an oral mucosa, trauma to an oral mucosa, periodontal disease, a cutaneous ulcer, or a skin scar afflicting said subject.

9 . The method of claim 8 wherein the skin scar is an acne scar.

10 . The method of claim 8 wherein the defect is increased depth of the subject's nasolabial groove.

11 . A method of stimulating collagen production comprising adding a substance P analog to dermal fibroblast cells to an in vitro fibroblast cell culture media wherein said substance P analog is according to Formula (I):

Z 1 -Xaa 1 -Xaa 2 -Xaa 3 -Xaa 4 -Xaa 5 -Xaa 6 -Xaa 7 -Xaa 8 -Xaa 9 -

Xaa 10 -Xaa 11 -Z 2 (I)

or a pharmaceutically acceptable salt thereof, wherein:

Xaa 1 is Arg, Lys, 6-N methyllysine, or (6-N, 6-N) dimethyllysine;

Xaa 2 is Pro or Ala;

Xaa 3 is Lys, Arg, 6-N-methyllysine, or (6-N, 6-N) dimethyllysine;

Xaa 4 is Pro or Ala;

Xaa 5 is Gln or Asn;

Xaa 6 is Gln or Asn;

Xaa 7 is Tyr, Phe, or Phe substituted with chlorine at position 2, 3, or 4;

Xaa 8 is Tyr, Phe, or Phe substituted with chlorine at position 2, 3, or 4;

Xaa 9 is Gly, Pro, Ala, or N-methylglycine;

Xaa 10 is Leu, Val, Ile, Norleucine, Met, Met sulfoxide, Met sulfone, N-methylleucine, or N-methylvaline;

Xaa 1 is Met, Met sulfoxide, Met sulfone, or norleucine;

Z 1 is R 2 N— or RC(O)NR—;

Z 2 is —C(O)NR 2 or —C(O)OR or a salt thereof;

each R is independently R is H, (C 1 -C 6 ) alkyl, (C 1 -C 6 ) alkenyl, (C 1 -C 6 ) alkynyl, (C 5 -C 20 ) aryl, (C 6 -C 26 ) alkaryl, 5-20 membered heteroaryl, or 6-26 membered alkheteroaryl; and

each “-” between residues Xaa 1 through Xaa 11 independently designates an amide linkage, a substitute amide linkage or an isostere of an amide wherein the substance P analog is not substance P (SEQ ID NO.: 1).

12 . The method of claim 11 wherein

Xaa 1 is Arg;

Xaa 2 is Pro;

Xaa 3 is Lys;

Xaa 4 is Pro;

Xaa 5 is Gln;

Xaa 6 is Gln;

Xaa 7 is Tyr, Phe, or Phe substituted with chlorine at position 4;

Xaa 8 is Tyr, Phe, or Phe substituted with chlorine at position 4;

Xaa 9 is Gly, Pro, or N-methylglycine;

Xaa 10 is Leu; and

Xaa 11 is Met, Met sulfoxide, Met sulfone, or norleucine.

13 . The method of claim 11 wherein the “-” between residues Xaa 1 through Xaa 11 designates

—C(O)NH—;

Z 1 is H 2 N—; and Z 2 is —C(O)NH 2 .

14 . The method of claim 11 wherein the substance P analog is selected from the group consisting of:

RPKPQQFFGLNle;

(SEQ ID NO.:2)

RPKPQQFFPLM;

(SEQ ID NO.:3)

RPKPQQFFMeGlyLM;

(SEQ ID NO.:4)

RPKPQQFTGLM;

(SEQ ID NO.:5)

RPKPQQF(4-Cl)F(4-Cl)GLM;

(SEQ ID NO.:6)

RPKPQQFFGLM(O);

(SEQ ID NO.:7)

RPKPQQFFMeGlyLM(O);

(SEQ ID NO.:8)

RPKPQQFFMeGLM(O 2 );

(SEQ ID NO.:9)

and

RPKPQQFFMeGlyLM(O 2 ).

(SEQ ID NO.:10)

15 . The method of claim 11 wherein the substance P analog is

Z 1 -RPKPQQFFMeGlyLM(O 2 )-Z 2 ;

wherein

Z 1 is NH 2 and Z 2 is C(O)NH 2 .

16 . The method of claim 11 wherein the fibroblast cells are human fibroblast cells.

17 . The method of claim 11 wherein the fibroblast cells are dermal fibroblast cells.

18 . The method of claim 11 , further comprising administering said fibroblast cells to a subject to treat a defect of an oral mucosa, trauma to an oral mucosa, periodontal disease, a cutaneous ulcer, or a skin scar afflicting said subject.

19 . The method of claim 18 wherein the skin scar is an acne scar.

20 . The method of claim 18 wherein the defect is increased depth of the subject's nasolabial groove.

21 . The method of claim 11 further comprising administering the fibroblast cells as a skin graft to a subject in need thereof.

22 . A composition for stimulating or promoting fibroblast cell proliferation comprising a substance P analog according to Formula (I)

Z 1 -Xaa 1 -Xaa 2 -Xaa 3 -Xaa 4 -Xaa 5 -Xaa 6 -Xaa 7 -Xaa 8 -Xaa 9 -

Xaa 10 -Xaa 11 -Z 2 (I)

or a pharmaceutically acceptable salt thereof, wherein:

Xaa 1 is Arg, Lys, 6-N methyllysine, or (6-N, 6-N) dimethyllysine;

Xaa 2 is Pro or Ala;

Xaa 3 is Lys, Arg, 6-N-methyllysine, or (6-N, 6-N) dimethyllysine;

Xaa 4 is Pro or Ala;

Xaa 5 is Gln or Asn;

Xaa 6 is Gln or Asn;

Xaa 7 is Tyr, Phe, or Phe substituted with chlorine at position 2, 3, or 4;

Xaa 8 is Tyr, Phe, or Phe substituted with chlorine at position 2, 3, or 4;

Xaa 9 is Gly, Pro, Ala, or N-methylglycine;

Xaa 10 is Leu, Val, Ile, norleucine, Met, Met sulfoxide, Met sulfone, N-methylleucine, or N-methylvaline;

Xaa 11 is Met, Met sulfoxide, Met sulfone, or norleucine;

Z 1 is R 2 N— or RC(O)NR—;

Z 2 is —C(O)NR 2 or —C(O)OR or a salt thereof;

each R is independently R is —H, (C 1 -C 6 ) alkyl, (C 1 -C 6 ) alkenyl, (C 1 -C 6 ) alkynyl, (C 5 -C 20 ) aryl, (C 6 -C 26 ) alkaryl, 5-20 membered heteroaryl, or 6-26 membered alkheteroaryl; and

each “-” between residues Xaa 1 through Xaa 11 independently designates an amide linkage, a substitute amide linkage or an isostere of an amide, wherein the substance P analog is not substance P.

23 . The composition of claim 22 wherein

Xaa 1 is Arg;

Xaa 2 is Pro;

Xaa 3 is Lys;

Xaa 4 is Pro;

Xaa 5 is Gln;

Xaa 6 is Gln;

Xaa 7 is Tyr, Phe, or Phe substituted with chlorine at position 4;

Xaa 8 is Tyr, Phe, or Phe substituted with chlorine at position 4;

Xaa 9 is Gly, Pro or N-methylglycine;

Xaa 10 is Leu; and

Xaa 11 is Met, Met sulfoxide, Met sulfone or Norleucine.

24 . The composition of claim 22 wherein the “-” between residues Xaa 1 through Xaa 11 designates

—C(O)NH—;

Z 1 is H 2 N—; and Z 2 is —C(O)NH 2 .

25 . The composition of claim 22 wherein the substance P analog is selected from the group consisting of:

RPKPQQFFGLNle;

(SEQ ID NO.:2)

RPKPQQFFPLM;

(SEQ ID NO.:3)

RPKPQQFFMeGlyLM;

(SEQ ID NO.:4)

RPKPQQFTGLM;

(SEQ ID NO.:5)

RPKPQQF(4-Cl)F(4-Cl)GLM;

(SEQ ID NO.:6)

RPKPQQFFGLM(O);

(SEQ ID NO.:7)

RPKPQQFFMeGlyLM(O);

(SEQ ID NO.:8)

RPKPQQFFMeGLM(O 2 );

(SEQ ID NO.:9)

and

RPKPQQFFMeGlyLM(O 2 ).

(SEQ ID NO.:10)

26 . The composition of claim 18 wherein the substance P analog is

Z 1 -RPKPQQFFMeGlyLM(O 2 )-Z 2 ;

wherein Z 1 is NH 2 and Z 2 is C(O)NH 2 .

27 . The composition of claim 22 wherein the fibroblast cells are human fibroblast cells.

28 . The composition of claim 22 wherein the fibroblast cells are dermal fibroblast cells.