Small molecule pipecolic acid derivative hair growth compositions and uses

This invention relates to pharmaceutical compositions and methods for treating alopecia and promoting hair growth using pipecolic acid derivatives.

BACKGROUND OF THE INVENTION
 1. Field of Invention
 This invention relates to pharmaceutical compositions and methods for
 treating alopecia and promoting hair growth using low molecular weight,
 small molecule pipecolic acid derivatives.
 2. Description of Related Art
 Hair loss occurs in a variety of situations. These situations include male
 pattern alopecia, alopecia senilis, alopecia areata, diseases accompanied
 by basic skin lesions or tumors, and systematic disorders such as
 nutritional disorders and internal secretion disorders. The mechanisms
 causing hair loss are very complicated, but in some instances can be
 attributed to aging, genetic disposition, the activation of male hormones,
 the loss of blood supply to hair follicles, and scalp abnormalities.
 The immunosuppressant drugs FK506, rapamycin and cyclosporin are well known
 as potent T-cell specific immunosuppressants, and are effective against
 graft rejection after organ transplantation. It has been reported that
 topical, but not oral, application of FK506 (Yamamoto et al., J. Invest.
 Dermatol., 1994, 102, 160-164; Jiang et al., J. Invest. Dermatol. 1995,
 104, 523-525) and cyclosporin (Iwabuchi et al., J. Dermatol. Sci. 1995, 9,
 64-69) stimulates hair growth in a dose-dependent manner. One form of hair
 loss, alopecia areata, is known to be associated with autoimmune
 activities; hence, topically administered immunomodulatory compounds are
 expected to demonstrate efficacy for treating that type of hair loss. The
 hair growth stimulating effects of FK506 have been the subject of an
 international patent filing covering FK506 and structures related thereto
 for hair growth stimulation (Honbo et al., EP 0 423 714 A2). Honbo et al.
 discloses the use of relatively large tricyclic compounds, known for their
 immunosuppressive effects, as hair revitalizing agents.
 The hair growth and revitalization effects of FK506 and related agents are
 disclosed in many U.S. patents (Goulet et al., U.S. Pat. No. 5,258,389;
 Luly et al., U.S. Pat. No. 5,457,111; Goulet et al., U.S. Pat. No.
 5,532,248; Goulet et al., U.S. Pat. No. 5,189,042; and Ok et al., U.S.
 Pat. No. 5,208,241; Rupprecht et al., U.S. Pat. No. 5,284,840; Organ et
 al., U.S. Pat. No. 5,284,877). These patents claim FK506 related
 compounds. Although they do not claim methods of hair revitalization, they
 disclose the known use of FK506 for effecting hair growth. Similar to
 FK506 (and the claimed variations in the Honbo et al. patent), the
 compounds claimed in these patents are relatively large. Further, the
 cited patents relate to immunomodulatory compounds for use in autoimmune
 related diseases, for which FK506's efficacy is well known.
 Other U.S. patents disclose the use of cyclosporin and related compounds
 for hair revitalization (Hauer et al., U.S. Pat. No. 5,342,625; Eberle,
 U.S. Pat. No. 5,284,826; Hewitt et al., U.S. Pat. No. 4,996,193). These
 patents also relate to compounds useful for treating autoimmune diseases
 and cite the known use of cyclosporin and related immunosuppressive
 compounds for hair growth.
 However, immunosuppressive compounds by definition suppress the immune
 system and also exhibit other toxic side effects. Accordingly, there is a
 need for non-immunosuppressant, small molecule compounds which are useful
 as hair revitalizing compounds.
 Hamilton and Steiner disclose in U.S. Pat. No. 5,614,547 novel pyrrolidine
 carboxylate compounds which bind to the immunophilin FKBP12 and stimulate
 nerve growth, but which lack immunosuppressive effects. Unexpectedly, it
 has been discovered that these non-immunosuppressant compounds promote
 hair growth with an efficacy similar to FK506. Yet their novel small
 molecule structure and non-immunosuppressive properties differentiate them
 from FK506 and related immunosuppressive compounds found in the prior art.
 SUMMARY OF THE INVENTION
 The present invention relates to a method for treating alopecia or
 promoting hair growth in an animal, which comprises administering to said
 animal an effective amount of a low molecular weight, small molecule
 pipecolic acid derivative.
 The present invention further relates to a pharmaceutical composition which
 comprises:
 (i) an effective amount of a pipecolic acid derivative for treating
 alopecia or promoting hair growth in an animal; and
 (ii) a pharmaceutically acceptable carrier.
 The pipecolic acid derivatives used in the inventive methods and
 pharmaceutical compositions include immunosuppressive and
 non-immunosuppressive compounds having an affinity for FKBP-type
 immunophilins, particularly FKBP12. Non-immunosuppressive compounds, as
 their name suggests, do not exert any significant immunosuppressive
 activity.

DETAILED DESCRIPTION OF THE INVENTION
 Definitions
 "Alopecia" refers to deficient hair growth and partial or complete loss of
 hair, including without limitation androgenic alopecia (male pattern
 baldness), toxic alopecia, alopecia senilis, alopecia areata, alopecia
 pelada and trichotillomania. Alopecia results when the pilar cycle is
 disturbed. The most frequent phenomenon is a shortening of the hair growth
 or anagen phase due to cessation of cell proliferation. This results in an
 early onset of the catagen phase, and consequently a large number of hairs
 in the telogen phase during which the follicles are detached from the
 dermal papillae, and the hairs fall out. Alopecia has a number of
 etiologies, including genetic factors, aging, local and systemic diseases,
 febrile conditions, mental stresses, hormonal problems, and secondary
 effects of drugs.
 "GPI 1044" refers to Compound 4.
 "GPI 1102" refers to 4-phenyl-1-(3-phenylpropyl) butyl 1-
 (3,3-dimethyl-2-oxopentanoyl) -2-piperidinecarboxylate.
 "GPI 1116" refers to 1-phenethyl-3-phenylpropyl
 1-(3,3-dimethyl-2-oxopentanoyl)-2-piperidinecarboxylate.
 "GPI 1206" refers to a compound of formula
 ##STR1##
 "Isomers" refer to different compounds that have the same molecular
 formula. "Stereoisomers" are isomers that differ only in the way the atoms
 are arranged in space. "Enantiomers" are a pair of stereoisomers that are
 non-superimposable mirror images of each other. "Diastereoisomers" are
 stereoisomers which are not mirror images of each other. "Racemic mixture"
 means a mixture containing equal parts of individual enantiomers.
 "Non-racemic mixture" is a mixture containing unequal parts of individual
 enantiomers or stereoisomers.
 "Pharmaceutically acceptable salt, ester, or solvate" refers to a salt,
 ester, or solvate of a subject compound which possesses the desired is
 pharmacological activity and which is neither biologically nor otherwise
 undesirable. A salt, ester, or solvate can be formed with inorganic acids
 such as acetate, adipate, alginate, aspartate, benzoate, benzenesulfonate,
 bisulfate, butyrate, citrate, camphorate, camphorsulfonate,
 cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate,
 fumarate, glucoheptanoate, gluconate, glycerophosphate, hemisulfate,
 heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide,
 2-hydroxyethanesulfonate, lactate, maleate, methanesulfonate, naphthylate,
 2-naphthalenesulfonate, nicotinate, oxalate, sulfate, thiocyanate,
 tosylate and undecanoate. Examples of base salts, esters, or solvates
 include ammonium salts; alkali metal salts, such as sodium and potassium
 salts; alkaline earth metal salts, such as calcium and magnesium salts;
 salts with organic bases, such as dicyclohexylamine salts;
 N-methyl-D-glucamine; and salts with amino acids, such as arginine,
 lysine, and so forth. Also, the basic nitrogen-containing groups can be
 quarternized with such agents as lower alkyl halides, such as methyl,
 ethyl, propyl, and butyl chlorides, bromides, and iodides; dialkyl
 sulfates, such as dimethyl, diethyl, dibutyl, and diamyl sulfates; long
 chain halides, such as decyl, lauryl, myristyl, and stearyl chlorides,
 bromides, and iodides; aralkyl halides, such as benzyl and phenethyl
 bromides; and others. Water or oil-soluble or dispersible products are
 thereby obtained.
 "Pilar cycle" refers to the life cycle of hair follicles, and includes
 three phases:
 (1) the anagen phase, the period of active hair growth which, insofar as
 scalp hair is concerned, lasts about three to five years;
 (2) the catagen phase, the period when growth stops and the follicle
 atrophies which, insofar as scalp hair is concerned, lasts about one to
 two weeks; and
 (3) the telogen phase, the rest period when hair progressively separates
 and finally falls out which, insofar as scalp hair is concerned, lasts
 about three to four months.
 Normally 80 to 90 percent of the follicles are in the anagen phase, less
 than 1 percent being in the catagen phase, and the rest being in the
 telogen phase. In the telogen phase, hair is uniform in diameter with a
 slightly bulbous, non-pigmented root. By contrast, in the anagen phase,
 hair has a large colored bulb at its root.
 "Promoting hair growth" refers to maintaining, inducing, stimulating,
 accelerating, or revitalizing the germination of hair.
 "Treating alopecia" refers to:
 (i) preventing alopecia in an animal which may be predisposed to alopecia;
 and/or
 (ii) inhibiting, retarding or reducing alopecia; and/or
 (iii) promoting hair growth; and/or
 (iv) prolonging the anagen phase of the hair cycle; and/or
 (v) converting vellus hair to growth as terminal hair. Terminal hair is
 coarse, pigmented, long hair in which the bulb of the hair follicle is
 seated deep in the dermis. Vellus hair, on the other hand, is fine, thin,
 non-pigmented short hair in which the hair bulb is located superficially
 in the dermis. As alopecia progresses, the hairs change from the terminal
 to the vellus type.
 METHODS OF THE PRESENT INVENTION
 The present invention relates to a method for treating alopecia or
 promoting hair growth in an animal, which comprises administering to said
 animal an effective amount of a pipecolic acid derivative.
 The inventive method is particularly useful for treating male pattern
 alopecia, alopecia senilis, alopecia areata, alopecia resulting from skin
 lesions or tumors, alopecia resulting from cancer therapy such as
 chemotherapy and radiation, and alopecia resulting from systematic
 disorders such as nutritional disorders and internal secretion disorders.
 Pharmaceutical Compositions of the Present Invention
 The present invention also relates to a pharmaceutical composition
 comprising:
 (i) an effective amount of a pipecolic acid derivative for treating
 alopecia or promoting hair growth in an animal; and
 (ii) a pharmaceutically acceptable carrier.
 PIPECOLIC ACID DERIVATIVES
 The pipecolic acid derivatives used in the methods and pharmaceutical
 compositions of the present invention are low molecular weight, small
 molecule compounds having an affinity for FKBP-type immunophilins, such as
 FKBP12. When a pipecolic acid derivative binds to an FKBP-type
 immunophilin, it has been found to inhibit the prolyl-peptidyl cis-trans
 isomerase, or rotamase, activity of the binding protein. Unexpectedly, the
 compounds have also been found to stimulate hair growth. These rotamase
 inhibiting compounds may be immunosuppressive or non-immunosuppressive.
 Examples of useful compounds are set forth below.
 FORMULA I
 An exemplary pipecolic acid derivative is a compound of formula I
 ##STR2##
 or a pharmaceutically acceptable salt, ester, or solvate thereof, wherein:
 m is 0-3;
 A is CH.sub.2, O, NH, or N--(C.sub.1 -C.sub.4 alkyl);
 B and D are independently Ar, C.sub.5 -C.sub.7 cycloalkyl substituted
 C.sub.1 -C.sub.6 straight or branched chain alkyl or C.sub.2 -C.sub.6
 straight or branched chain alkenyl, C.sub.5 -C.sub.7 cycloalkenyl
 substituted C.sub.1 -C.sub.6 straight or branched chain alkyl or C.sub.2
 -C.sub.6 straight or branched chain alkenyl, or Ar substituted C.sub.1
 -C.sub.6 straight or branched chain alkyl or C.sub.2 -C.sub.6 straight or
 branched chain alkenyl, wherein in each case, one or two carbon atom(s) of
 said alkyl or alkenyl may be substituted with one or two heteroatom(s)
 independently selected from the group consisting of oxygen, sulfur, So,
 and SO.sub.2 in chemically reasonable substitution patterns, or
 ##STR3##
 wherein Q is hydrogen, C.sub.1 -C.sub.6 straight or branched chain alkyl,
 or C.sub.2 -C.sub.6 straight or branched chain alkenyl; and
 T is Ar or C.sub.5 -C.sub.7 cycloalkyl substituted at positions 3 and 4
 with substituents independently selected from the group consisting of
 hydrogen, hydroxy, O--(C.sub.1 -C.sub.4 alkyl), O--(C.sub.2 -C.sub.4
 alkenyl), and carbonyl;
 Ar is selected from the group consisting of 1-napthyl, 2-napthyl, 2-furyl,
 3-furyl, 2-thienyl, 3-thienyl, 2-pyridyl, 3-pyridyl, 4-pyridyl and phenyl,
 monocyclic and bicyclic heterocyclic ring systems with individual ring
 sizes being 5 or 6 which contain in either or both rings a total of 1-4
 heteroatom(s) independently selected from the group consisting of oxygen,
 nitrogen and sulfur; wherein Ar contains 1-3 substituent(s) independently
 selected from the group consisting of hydrogen, halo, hydroxy,
 hydroxymethyl, nitro, CF.sub.3, trifluoromethoxy, C.sub.1 -C.sub.6
 straight or branched chain alkyl, C.sub.2 -C.sub.6 straight or branched
 chain alkenyl, O--(C.sub.1 -C.sub.4 straight or branched chain alkyl),
 O--(C.sub.2 -C.sub.4 straight or branched chain alkenyl), O-benzyl,
 O-phenyl, amino, 1,2-methylenedioxy, carbonyl, and phenyl;
 L is either hydrogen or U; M is either oxygen or CH--U, provided that if L
 is hydrogen, then M is CH--U, or if M is oxygen then L is U;
 U is hydrogen, O--(C.sub.1 -C.sub.4 straight or branched chain alkyl),
 O--(C.sub.2 -C.sub.4 straight or branched chain alkenyl), C.sub.1 -C.sub.6
 straight or branched chain alkyl, C.sub.2 -C.sub.6 straight is or branched
 chain alkenyl, C.sub.5 -C.sub.7 cycloalkyl, C.sub.5 -C.sub.7 cycloalkenyl
 substituted with C.sub.1 -C.sub.4 straight or branched chain alkyl or
 C.sub.2 -C.sub.4 straight or branched chain alkenyl, (C.sub.1 -C.sub.4
 alkyl or C.sub.2 -C.sub.4 alkenyl)-Ar, or Ar;
 J is hydrogen, C.sub.1 or C.sub.2 alkyl, or benzyl; K is C.sub.1 -C.sub.4
 straight or branched chain alkyl, benzyl or cyclohexylmethyl; or J and K
 are taken together to form a 5-7 membered heterocyclic ring which is
 substituted with oxygen, sulfur, SO, or SO.sub.2 ; and
 said pipecolic acid derivative has an affinity for FKBP-type immunophilins.
 Representative species of Formula I are presented in Table I.
 TABLE I
 ##STR4##
 Compound n m B D L
 1 2 0 3-Phenyl- 3-(3-Pyridyl)- Phenyl
 propyl propyl
 2 2 0 3-Phenyl- 3-(2-Pyridyl)- Phenyl
 propyl propyl
 3 2 0 3-Phenyl- 2-(4-Methoxy- Phenyl
 propyl phenyl)ethyl
 4 2 0 3-Phenyl- 3-Phenylpropyl Phenyl
 propyl
 5 2 0 3-Phenyl- 3-Phenylpropyl 3,4,5-
 propyl Trimeth-
 oxyphenyl
 6 2 0 3-Phenyl- 2-(3-Pyridyl)- 3,4,5-
 propyl propyl Trimeth-
 oxyphenyl
 7 2 0 3-Phenyl- 3-(2-Pyridyl)- 3,4,5-
 propyl propyl Trimeth-
 oxyphenyl
 8 2 0 3-Phenyl- 3-(4-Methoxy- 3,4,5-
 propyl phenyl)propyl Trimeth-
 oxyphenyl
 9 2 0 3-Phenyl- 3-(3-Pyridyl)- 3-Iso-
 propyl propyl propoxy-
 phenyl
 FORMULA II
 U.S. Pat. No. 5,330,993, incorporated herein by reference, discloses an
 exemplary pipecolic acid derivative of Formula II
 ##STR5##
 or a pharmaceutically acceptable salt, ester, or solvate thereof, wherein:
 A is O, NH, or N--(C.sub.1 -C.sub.4 alkyl);
 B is hydrogen, CHL--Ar, C.sub.1 -C.sub.6 straight or branched chain alkyl,
 C.sub.2 -C.sub.6 straight or branched chain alkenyl, C.sub.5 -C.sub.7
 cycloalkyl, C.sub.5 -C.sub.7 cycloalkenyl, Ar substituted C.sub.1 -C.sub.6
 alkyl or C.sub.2 -C.sub.6 alkenyl, or
 ##STR6##
 wherein L and Q are independently hydrogen, C.sub.1 -C.sub.6 straight or
 branched chain alkyl, or C.sub.2 -C.sub.6 straight or branched chain
 alkenyl; and
 T is Ar or C.sub.5 -C.sub.7 cyclohexyl substituted at positions 3 and 4
 with substituents independently selected from the group consisting of
 hydrogen, hydroxy, O--(C.sub.1 -C.sub.4 alkyl), O--(C.sub.2 -C.sub.4
 alkenyl), and carbonyl;
 Ar is selected from the group consisting of 1-napthyl, 2-napthyl, 2-furyl,
 3-furyl, 2-thienyl, 2-pyridyl, 3-pyridyl, 4-pyridyl and phenyl having 1-3
 substituent(s) independently selected from the group consisting of
 hydrogen, halo, hydroxy, nitro, CF.sub.3, C.sub.1 -C.sub.6 straight or
 branched chain alkyl, C.sub.2 -C.sub.6 straight or branched chain alkenyl,
 O--(C.sub.1 -C.sub.4 straight or branched chain alkyl), O--(C.sub.2
 -C.sub.4 straight or branched chain alkenyl), O-benzyl, O-phenyl, amino,
 and phenyl.
 D is hydrogen or U; E is oxygen or CH--U, provided that if D is hydrogen,
 then E is CH--U, or if E is oxygen, then D is U;
 U is hydrogen, O--(C.sub.1 -C.sub.4 straight or branched chain alkyl),
 O--(C.sub.2 -C.sub.4 straight or branched chain alkenyl), C.sub.1 -C.sub.6
 straight or branched chain alkyl, C.sub.2 -C.sub.6 straight or branched
 chain alkenyl, C.sub.5 -C.sub.7 -cycloalkyl, C.sub.5 -C.sub.7 cycloalkenyl
 substituted with C.sub.1 -C.sub.4 straight or branched chain alkyl or
 C.sub.2 -C.sub.4 straight or branched chain alkenyl, 2-indolyl, 3-indolyl,
 (C.sub.1 -C.sub.4 alkyl or C.sub.2 -C.sub.4 alkenyl)-Ar, or Ar;
 J is hydrogen, C.sub.1 or C.sub.2 alkyl, or benzyl; K is C.sub.1 -C.sub.4
 straight or branched chain alkyl, benzyl or cyclohexylethyl; or J and K
 are taken together to form a 5-7 membered heterocyclic ring which is
 substituted with oxygen, sulfur, SO, or SO.sub.2.
 FORMULA III
 A preferred pipecolic acid derivative is a compound of Formula III
 ##STR7##
 or a pharmaceutically acceptable salt, ester, or solvate thereof, wherein:
 n is 2;
 D is phenyl, methoxy, 2-furyl, or 3,4,5-trimethoxyphenyl; and
 B is benzyl, 3-phenylpropyl, 4-(4-methoxyphenyl)butyl, 4-phenylbutyl,
 phenethyl, 3-cyclohexylpropyl, 4-cyclohexylbutyl, 3-cyclopentylpropyl,
 4-cyclohexylbutyl, 3-phenoxybenzyl, 3-(3-indolyl)propyl, or
 4-(4-methoxyphenyl)butyl;
 provided that:
 when D is phenyl, then B is benzyl, 3-phenylpropyl,
 4-(4-methoxyphenyl)butyl, 4-phenylbutyl, phenethyl, or 4-cyclohexylbutyl;
 when D is methoxy, B is benzyl, 4-cyclohexylbutyl, 3-cyclohexylpropyl, or
 3-cyclopentylpropyl;
 when D is 2-furyl, then B is benzyl; and
 when D is 3,4,5-trimethoxyphenyl, then B is 4-cyclohexylbutyl,
 3-phenoxybenzyl, 4-phenylbutyl, 3- (3-indolyl) propyl, or 4-
 (4-methoxyphenyl) butyl.
 Representative species of Formula III are presented in Table II.
 TABLE II
 III
 ##STR8##
 Compound B D n
 10 Benzyl Phenyl 2
 11 3-Phenylpropyl Phenyl 2
 12 4-(4-Methoxyphenyl)- Phenyl 2
 butyl
 13 4-Phenylbutyl Phenyl 2
 14 Phenethyl Phenyl 2
 15 4-Cyclohexylbutyl Phenyl 2
 16 Benzyl Methoxy 2
 17 4-Cyclohexylbutyl Methoxy 2
 18 3-Cyclohexylpropyl Methoxy 2
 19 3-Cyclopentylpropyl Methoxy 2
 20 Benzyl 2-Furyl 2
 21 4-Cyclohexylbutyl 3,4,5- 2
 Trimethoxyphenyl
 22 3-Phenoxybenzyl 3,4,5- 2
 Trimethoxyphenyl
 23 4-Phenylbutyl 3,4,5- 2
 Trimethoxyphenyl
 24 3-(3-Indolyl)propyl 3,4,5- 2
 Trimethoxyphenyl
 25 4-(4-Methoxyphenyl)- 3,4,5- 2
 butyl Trimethoxyphenyl
 FORMULA IV
 The pipecolic acid derivative may also be a compound of formula IV
 ##STR9##
 or a pharmaceutically acceptable salt, ester, or solvate thereof, wherein:
 V is C, N, or S;
 J and K, taken together with V and the carbon atom to which they are
 respectively attached, form a 5-7 membered saturated or unsaturated
 heterocyclic ring containing, in addition to V, one or more heteroatom(s)
 selected from the group consisting of O, S, SO, SO.sub.2, N, NH, and NR;
 R is either C.sub.1 -C.sub.9 straight or branched chain alkyl, C.sub.2
 -C.sub.9 straight or branched chain alkenyl, C.sub.3 -C.sub.9 cycloakyl,
 C.sub.5 -C.sub.7 cycloalkenyl, or Ar.sub.1, wherein R is either
 unsubstituted of substituted with one or more substituent(s) independently
 selected from the group consisting of halo, haloalkyl, carbonyl, carboxy,
 hydroxy, nitro, trifluoromethyl, C.sub.1 -C.sub.6 straight or branched
 chain alkyl, C.sub.2 -C.sub.6 straight or branched chain alkenyl, C.sub.1
 -C.sub.4 alkoxy, C.sub.2 -C.sub.4 alkenyloxy, phenoxy, benzyloxy,
 thioalkyl, alkylthio, sulfhydryl, amino, alkylamino, aminoalkyl,
 aminocarboxyl, and Ar.sub.2 ;
 Ar.sub.1 and Ar.sub.2 are independently an alicyclic or aromatic, mono-,
 bi- or tricyclic, carbo- or heterocyclic ring; wherein the individual ring
 size is 5-8 members; wherein said heterocyclic ring contains 1-6
 heteroatom(s) independently selected from the group consisting of O, N,
 and S;
 A, B, D, L, M, and m are as defined in Formula I above; and
 said pipecolic acid derivative has an affinity for FKBP-type immunophilins.
 All the compounds of Formulas I-IV possess asymmetric centers and thus can
 be produced as mixtures of stereoisomers or as individual R- and
 S-stereoisomers. The individual stereoisomers may be obtained by using an
 optically active starting material, by resolving a racemic or non-racemic
 mixture of an intermediate at some appropriate stage of the synthesis, or
 by resolving the compounds of Formulas I-IV. It is understood that the
 compounds of Formulas I-IV encompass individual stereoisomers as well as
 mixtures (racemic and non-racemic) of stereoisomers. Preferably,
 S-stereoisomers are used in the pharmaceutical compositions and methods of
 the present invention.
 Affinity for FKBP12
 The compounds used in the inventive methods and pharmaceutical compositions
 have an affinity for the FK506 binding protein, particularly FKBP12. The
 inhibition of the prolyl peptidyl cis-trans isomerase activity of FKBP may
 be measured as an indicator of this affinity.
 K. Test Procedure
 Inhibition of the peptidyl-prolyl isomerase (rotamase) activity of the
 compounds used in the inventive methods and pharmaceutical compositions
 can be evaluated by known methods described in the literature (Harding et
 al., Nature, 1989, 341:758-760; Holt et al. J. Am. Chem. Soc.,
 115:9923-9938). These values are obtained as apparent K.sub.i 's and are
 presented for representative compounds in TABLE III.
 The cis-trans isomerization of an alanine-proline bond in a model
 substrate, N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide, is monitored
 spectrophotometrically in a chymotrypsin-coupled assay, which releases
 para-nitroanilide from the trans form of the substrate.
 The inhibition of this reaction caused by the addition of different
 concentrations of inhibitor is determined, and the data is analyzed as a
 change in first-order rate constant as a function of inhibitor
 concentration to yield the apparent K.sub.i values.
 In a plastic cuvette are added 950 mL of ice cold assay buffer (25 mM
 HEPES, pH 7.8, 100 mM NaCl), 10 mL of FKBP (2.5 mM in 10 mM Tris-Cl pH
 7.5, 100 mM NaCl, 1 mM dithiothreitol), 25 mL of chymotrypsin (50 mg/ml in
 1 mM HCl) and 10 mL of test compound at various concentrations in dimethyl
 sulfoxide. The reaction is initiated by the addition of 5 mL of substrate
 (succinyl-Ala-Phe-Pro-Phe-para-nitroanilide, 5 mg/mL in 2.35 mM LiCl in
 trifluoroethanol).
 The absorbance at 390 nm versus time is monitored for 90 seconds using a
 spectrophotometer and the rate constants are determined from the
 absorbance versus time data files.
 TABLE III
 In Vitro Test Results - Formulas I-III
 Compound K.sub.i (.mu.M)
 10 1.5
 13 0.35
 14 1.1
 15 0.4
 16 80
 17 6
 18 20
 19 35
 20 3
 21 0.04
 22 0.018
 23 0.019
 24 0.017
 25 0.013
 Route of Administration
 To effectively treat alopecia or promote hair growth, the compounds used in
 the inventive methods and pharmaceutical compositions must readily affect
 the targeted areas. For these purposes, the compounds are preferably
 administered topically to the skin.
 For topical application to the skin, the compounds can be formulated into
 suitable ointments containing the compounds suspended or dissolved in, for
 example, mixtures with one or more of the following: mineral oil, liquid
 petrolatum, white petrolatum, propylene glycol, polyoxyethylene
 polyoxypropylene compound, emulsifying wax and water. Alternatively, the
 compounds can be formulated into suitable lotions or creams containing the
 active compound suspended or dissolved in, for example, a mixture of one
 or more of the following: mineral oil, sorbitan monostearate, polysorbate
 60, cetyl ester wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol
 and water.
 Other routes of administration known in the pharmaceutical art are also
 contemplated by this invention.
 Dosage
 Dosage levels on the order of about 0.1 mg to about 10,000 mg of the active
 ingredient compound are useful in the treatment of the above conditions,
 with preferred levels of about 0.1 mg to about 1,000 mg. The specific dose
 level for any particular patient will vary depending upon a variety of
 factors, including the activity of the specific compound employed; the
 age, body weight, general health, sex and diet of the patient; the time of
 administration; the rate of excretion; drug combination; the severity of
 the particular disease being treated; and the form of administration.
 Typically, in vitro dosage-effect results provide useful guidance on the
 proper doses for patient administration. Studies in animal models are also
 helpful. The considerations for determining the proper dose levels are
 well known in the art.
 The compounds can be administered with other hair revitalizing agents.
 Specific dose levels for the other hair revitalizing agents will depend
 upon the factors previously stated and the effectiveness of the drug
 combination.
 EXAMPLES
 The following examples are illustrative of the present invention and are
 not intended to be limitations thereon. Unless otherwise indicated, all
 percentages are based upon 100% by weight of the final composition.
 Example 1
 In Vivo Hair Generation Tests With C57 Black 6 Mice
 Experiment A: C57 black 6 mice were used to demonstrate the hair
 revitalizing properties of pipecolic acid derivative GPI 1044 (compound
 4), as well as related pipecolic acid derivative neuroimmunophilin FKBP
 ligands GPI 1102 and GPI 1116. C57 black 6 mice, approximately 7 weeks
 old, had an area of about 2 inches by 2 inches on their hindquarters
 shaved to remove all existing hair. Care was taken not to nick or cause
 abrasion to the underlaying dermal layers. The animals were in anagen
 growth phase, as indicated by the pinkish color of the skin. Referring now
 to FIGS. 1 and 2, four animals were treated by topical administration with
 20% propylene glycol vehicle (FIG. 1), and seven animals were treated by
 topical administration with 10 .mu.M GPI 1044 (FIG. 2). The animals were
 treated with vehicle or GPI 1044 every 48 hours (3 applications total over
 the course of 5 days) and the hair growth was allowed to proceed for 6
 weeks. Hair growth was quantitated by the percent of shaved area covered
 by new hair growth during this time period.
 FIG. 1 shows that animals treated with vehicle exhibited only a small
 amount of hair growth in patches or tufts, with less than 3% of the shaved
 area covered with new growth. In contrast, FIG. 2 shows that animals
 treated with 10 .mu.M GPI 1044 exhibited dramatic hair growth, covering as
 much as 50% of the shaved area in some animals. FIG. 3 compares the hair
 growth score of unshaven animals with the hair growth scores of shaven
 animals treated with a vehicle and GPI 1044 (1 .mu.M, 3 .mu.M and 10
 .mu.M), as well as related neuroimmunophilin FKBP ligands GPI 1116 (1
 .mu.M and 10 .mu.M) and GPI 1102 (1 .mu.M and 3 .mu.M).
 Experiment B: C57 Black 6 mice were used to demonstrate the hair
 revitalizing properties of neuroimmunophilin FKBP ligands. C57 Black 6
 mice, 55 to 75 days old, had an area of about 2 inches by 2 inches on
 their hindquarters shaved to remove all existing hair. Care was taken not
 to nick or cause abrasion to the underlying dermal layers. The animals
 were in a anagen growth phase when shaved. Five animals per group were
 treated by topical administration with a vehicle, FK506, or a
 neuroimmunophilin FKBP ligand (GPI 1116 or 1206) at a concentration of one
 micromole per milliliter to the shaved area. The animals were treated
 three times per week, and hair growth was evaluated 14 days after
 initiation of treatment. Hair growth was quantitated by the percent of
 shaved area covered by new hair growth, as scored by a blinded observer,
 on a scale of 0 (no growth) to five (complete hair regrowth in shaved
 area).
 FIG. 4 shows that after 14 days, the animals treated with vehicle exhibited
 the beginning of growth in small tufts. In contrast, animals treated with
 one of the low molecular weight, small molecule, neuroimmunophilin FKBP
 ligands exhibited dramatic hair growth.
 Example 2
 A lotion comprising the following composition may be prepared.

(%)
 95% Ethanol 80.0
 a pipecolic acid derivative as defined above 10.0
 .alpha.-Tocopherol acetate 0.01
 Ethylene oxide (40 mole) adducts of hardened 0.5
 castor oil
 purified water 9.0
 perfume and dye q.s.
 Into 95% ethanol are added a pipecolic acid derivative, .alpha.-tocopherol
 acetate, ethylene oxide (40 mole) adducts of hardened castor oil, perfume
 and a dye. The resulting mixture is stirred and dissolved, and purified
 water is added to the mixture to obtain a transparent liquid lotion.
 5 ml of the lotion may be applied once or twice per day to a site having
 marked baldness or alopecia.
 Example 3
 A lotion comprising the following composition shown may be prepared.

(%)
 95% Ethanol 80.0
 a pipecolic acid derivative as defined above 0.005
 Hinokitol 0.01
 Ethylene oxide (40 mole) adducts of hardened 0.5
 castor oil
 Purified water 19.0
 Perfume and dye q.s.
 Into 95% ethanol are added a pipecolic acid derivative, hinokitol, ethylene
 oxide (40 mole) adducts of hardened castor oil, perfume, and a dye. The
 resulting mixture is stirred, and purified water is added to the mixture
 to obtain a transparent liquid lotion.
 The lotion may be applied by spraying once to 4 times per day to a site
 having marked baldness or alopecia.
 Example 4
 An emulsion may be prepared from A phase and B phase having the following
 compositions.

(A phase) (%)
 Whale wax 0.5
 Cetanol 2.0
 Petrolatum 5.0
 Squalane 10.0
 Polyoxyethylene (10 mole) monostearate 2.0
 Sorbitan monooleate 1.0
 a pipecolic acid derivative as defined above 0.01
 (B phase) (%)
 Glycerine 10.0
 Purified water 69.0
 Perfume, dye, and preservative q.s.
 The A phase and the B phase are respectively heated and melted and
 maintained at 80.degree. c. Both phases are then mixed and cooled under
 stirring to normal temperature to obtain an emulsion.
 The emulsion may be applied by spraying once to four times per day to a
 site having marked baldness or alopecia.
 Example 5
 A cream may be prepared from A phase and B phase having the following
 compositions.

(A Phase) (%)
 Fluid paraffin 5.0
 Cetostearyl alcohol 5.5
 Petrolatum 5.5
 Glycerine monostearate 33.0
 Polyoxyethylene (20 mole) 2-octyldodecyl 3.0
 ether
 Propylparaben 0.3
 (B Phase) (%)
 a pipecolic acid derivative as defined above 0.8
 Glycerine 7.0
 Dipropylene glycol 20.0
 Polyethylene glycol 4000 5.0
 Sodium Hexametaphosphate 0.005
 Purified water 44.895
 The A phase is heated and melted, and maintained at 70.degree. C. The B
 phase is added into the A phase and the mixture is stirred to obtain an
 emulsion. The emulsion is then cooled to obtain a cream.
 The cream may be applied once to 4 times per day to a site having marked
 baldness or alopecia.
 Example 6
 A liquid comprising the following composition may be prepared.

(%)
 Polyoxyethylene butyl ether 20.0
 Ethanol 50.0
 a pipecolic acid derivative as defined above 0.001
 Propylene glycol 5.0
 Polyoxyethylene hardened castor oil 0.4
 derivative (ethylene oxide 80 mole adducts)
 Perfume q.s.
 Purified water q.s.
 Into ethanol are added polyoxypropylene butyl ether, propylene glycol,
 polyoxyethylene hardened castor oil, a pipecolic acid derivative, and
 perfume. The resulting mixture is stirred, and purified water is added to
 the mixture to obtain a liquid.
 The liquid may be applied once to 4 times per day to a site having marked
 baldness or alopecia.
 Example 7
 A shampoo comprising the following composition may be prepared.

(%)
 Sodium laurylsulfate 5.0
 Triethanolamine laurylsulfate 5.0
 Betaine lauryldimethylaminoacetate 6.0
 Ethylene glycol distearate 2.0
 Polyethylene glycol 5.0
 a pipecolic acid derivative as defined above 5.0
 Ethanol 2.0
 Perfume 0.3
 Purified water 69.7
 Into 69.7 of purified water are added 5.0 g of sodium laurylsulfate, 5.0 g
 of triethanolamine laurylsulfate, 6.0 g of betaine
 lauryldimethyl-aminoacetate. Then a mixture obtained by adding 5.0 g of a
 pipecolic acid derivative, 5.0 g of polyethylene glycol, and 2.0 g of
 ethylene glycol distearate to 2.0 g of ethanol, followed by stirring, and
 0.3 g of perfume are successively added. The resulting mixture is heated
 and subsequently cooled to obtain a shampoo.
 The shampoo may be used on the scalp once or twice per day.
 Example 8
 A patient is suffering from alopecia senilis. A pipecolic acid derivative,
 or a pharmaceutical composition comprising the same, may be administered
 to the patient. Increased hair growth is expected to occur following
 treatment.
 Example 9
 A patient is suffering from male pattern alopecia. A pipecolic acid
 derivative, or a pharmaceutical composition comprising the same, may be
 administered to the patient. Increased hair growth is expected to occur
 following treatment.
 Example 10
 A patient is suffering from alopecia areata. A pipecolic acid derivative,
 or a pharmaceutical composition comprising the same, may be administered
 to the patient. Increased hair growth is expected to occur following
 treatment.
 Example 11
 A patient is suffering from hair loss caused by skin lesions. A pipecolic
 acid derivative, or a pharmaceutical composition comprising the same, may
 be administered to the patient. Increased hair growth is expected to occur
 following treatment.
 Example 12
 A patient is suffering from hair loss caused by tumors. A pipecolic acid
 derivative, or a pharmaceutical composition comprising the same, may be
 administered to the patient. Increased hair growth is expected to occur
 following treatment.
 Example 13
 A patient is suffering from hair loss caused by a systematic disorder, such
 as a nutritional disorder or an internal secretion disorder. A pipecolic
 acid derivative, or a pharmaceutical composition comprising the same, may
 be administered to the patient. Increased hair growth is expected to occur
 following treatment.
 Example 14
 A patient is suffering from hair loss caused by chemotherapy. A pipecolic
 acid derivative, or a pharmaceutical composition comprising the same, may
 be administered to the patient. Increased hair growth is expected to occur
 following treatment.
 Example 15
 A patient is suffering from hair loss caused by radiation. A pipecolic acid
 derivative, or a pharmaceutical composition comprising the same, may be
 administered to the patient. Increased hair growth is expected to occur
 following treatment.
 The invention being thus described, it will be obvious that the same may be
 varied in many ways. Such variations are not to be regarded as a departure
 from the spirit and scope of the invention and all such modifications are
 intended to be included within the scope of the following claims.