U.S. patent application number 09/805347 was filed with the patent office on 2001-09-06 for method for inhibiting or treating chemotherapy-induced hair loss.
Invention is credited to Atwal, Karnail S..
Application Number | 20010020038 09/805347 |
Document ID | / |
Family ID | 27489728 |
Filed Date | 2001-09-06 |
United States Patent
Application |
20010020038 |
Kind Code |
A1 |
Atwal, Karnail S. |
September 6, 2001 |
Method for inhibiting or treating chemotherapy-induced hair
loss
Abstract
A method for inhibiting hair loss and/or promoting hair growth
in chemotherapy and/or radiation therapy patients wherein the
(R)-enantiomer of
4-[[(cyanoimino)-[(1,2,2-trimethylpropyl)amino]methyl]amino]benzonitri-
le is administered prior to, simultaneous with and/or after
chemotherapy and/or radiation treatment.
Inventors: |
Atwal, Karnail S.; (Newtown,
PA) |
Correspondence
Address: |
MARLA J MATHIAS
BRISTOL-MYERS SQUIBB COMPANY
PATENT DEPARTMENT
P O BOX 4000
PRINCETON
NJ
08543-4000
US
|
Family ID: |
27489728 |
Appl. No.: |
09/805347 |
Filed: |
March 13, 2001 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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09805347 |
Mar 13, 2001 |
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09447002 |
Nov 22, 1999 |
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09447002 |
Nov 22, 1999 |
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09119884 |
Jul 21, 1998 |
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6013668 |
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60055568 |
Aug 13, 1997 |
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60071364 |
Jan 15, 1998 |
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Current U.S.
Class: |
514/524 ;
514/171; 514/252.17; 514/284 |
Current CPC
Class: |
A61K 31/277 20130101;
A61K 8/43 20130101; C07C 279/28 20130101; C07B 2200/07 20130101;
A61Q 7/00 20130101 |
Class at
Publication: |
514/524 ;
514/171; 514/252.17; 514/284 |
International
Class: |
A61K 031/56; A61K
031/277; A61K 031/496 |
Claims
What is claimed is:
1. A method for preventing or inhibiting chemotherapy-induced hair
loss or radiation-induced hair loss or for promoting hair growth in
a patient having chemotherapy-induced hair loss or
radiation-induced hair loss, which comprises administering to a
human or other mammal a therapeutically effective amount of the
(R)-enantiomer of
4-[[(cyanoimino)[(1,2,2-trimethylpropyl)amino]methyl]amino]benzonitrile.
2. The method as defined in claim 1 wherein the (R)-enantiomer is
administered prior to, simultaneous with and/or subsequent to
chemotherapy and/or radiation therapy.
3. The method as defined in claim 1 wherein the (R)-enantiomer is
administered prior to chemotherapy and/or radiation therapy.
4. The method as defined in claim 1 wherein the (R)-enantiomer is
administered simultaneously with and/or subsequent to chemotherapy
and/or radiation therapy.
5. The method as defined in claim 1 wherein the (R)-enantiomer is
administered topically.
6. The method as defined in claim 1 wherein the (R)-enantiomer is
administered as a cream formulation, lotion formulation, liquid
formulation or ointment formulation.
7. The method as defined in claim 1 wherein the (R)-enantiomer is
administered systemically.
8. The method as defined in claim 1 for promoting hair growth in a
patient having chemotherapy-induced hair loss.
9. The method as defined in claim 1 for inhibiting
chemotherapy-induced hair loss.
10. The method as defined in claim 1 for promoting hair growth in a
patient having radiation-induced hair loss.
11. The method as defined in claim 1 for inhibiting
radiation-induced hair loss.
12. The method according to claim 1 wherein the (R)-enantiomer is
administered in conjunction with a chemotherapeutic agent which is
an antineoplastic agent selected from the group consisting of a
microtuble-stabilizing agent, a microtuble-disruptor agent, an
alkylating agent, an anti-metabolite, epidophyllotoxin, an
antineoplastic enzyme, a topoisomerase inhibitor, procarbazine,
mitoxantrone, a platinum coordination complex, a biological
response modifier, a growth inhibitor, a hormonal/antihormonal
therapeutic agent and a haematopoietic growth factor.
13. The method according to claim 1 wherein the (R)-enantiomer is
administered in conjunction with a chemotherapeutic agent which is
an antineoplastic agent selected from the group consisting of an
anthracycline drug, a vinca drug, a mitomycin, a bleomycin, a
cytotoxic nucleoside, a taxane, an epothilone, discodermolide, a
pteridine drug, a diynene, an aromatase inhibitor and a
podophyllotoxin.
14. The method according to claim 1 wherein the (R)-enantiomer is
administered in conjunction with a chemotherapeutic agent which is
an antineoplastic agent selected from the group consisting of
paclitaxel, docetaxel, 7-O-methylthiomethyl-paclitaxel,
3'-tert-butyl-3'-N-tert-butyl-
oxycarbonyl-4-deacetyl-3'-dephenyl-3'-N-debenzoyl-4-O-methoxycarbonyl-pacl-
itaxel, C-4 methyl carbonate paclitaxel, epothilone A, epothilone
B, epothilone C, epothilone D, desoxyepothilone A, desoxyepothilone
B, [1S-[1R*,3R*(E),7R*,10S*,11R*,12R*,
16S*]]-7,11-dihydroxy-8,8,10,12,16-pe-
ntamethyl-3-[1-methyl-2-(2-methyl-4-thiazolyl)ethenyl]-4-aza-17-oxabicyclo-
[ 14.1.0]heptadecase-5,9-dione,
[1S-[1R*,3R*(E),7R*,10S*,11R*,12R*,16S*]]--
3-[2-[2-(aminomethyl)-4-thiazolyl]-1-methylethenyl]-7,11-dihydroxy-8,8,10,-
12,16-pentamethyl-4,17-dioxabicyclo[14.1.0]heptadecane-5,9-dione,
doxorubicin, carminomycin, daunorubicin, aminopterin, methotrexate,
methopterin, dichloro-methotrexate, mitomycin C, porfiromycin,
5-fluorouracil, 6-mercaptopurine, gemcitabine, cytosine
arabinoside, podophyllotoxin, etoposide, etoposide phosphate,
teniposide, melphalan, vinblastine, vincristine, leurosidine,
vindesine, leurosine, estramustine, cisplatin, carboplatin,
cyclophosphamide, bleomycin, tamoxifen, ifosamide, hexamethyl
melamine, thiotepa, cytarabin, idatrexate, trimetrexate,
dacarbazine, L-asparaginase, camptothecin, CPT-11, topotecan,
ara-C, bicalutamide, flutamide, leuprolide, a pyridobenzoindole, an
interferon and an interleukin.
15. The method is defined in claim 1 wherein the (R)-enantiomer is
administered in the form of a topical formulation comprising
ethanol, propylene glycol and water.
16. The method as defined in claim 15 wherein the ethanol/propylene
glycol/water are in a 60/30/10 proportion.
17. The method as defined in claim 1 wherein the (R)-enantiomer is
administered in an amount from about 0.01 to about 15% by weight of
said (R)-enantiomer.
18. The method as defined in claim 1 wherein the (R)-enantiomer is
administered in the form of ethanol/propylene glycol/water,
60/30/10, in an amount to provide a 2% solution of the
(R)-enantiomer.
19. The method as defined in claim 1 wherein the (R)-enantiomer is
administered in combination with one or more other hair growth
promoting agents.
20. The method as defined in claim 19 wherein said other hair
growth promoting agent is another potassium channel opener, a
5-.alpha.-reductase inhibitor, an androgen blocker, betamethasone
dipropionate, a corticosteroid, scopolamine and/or cyproterone
acetate.
21. The method as defined in claim 20 wherein the other potassium
channel opener is minoxidil, diazoxide, cromakalim and/or
pinacidil; the 5-.alpha.-reductase inhibitor is finasteride,
terazosin HCl, and/or doxaosin mesylate; the androgen blocker is
4-(5-methoxyheptyl)-hexahydro-- 2(1H)-pentalenone; and the
corticosteroid is hydrocortisone.
22. The method as defined in claim 21 wherein said other hair
growth promoting agent is a 5-.alpha.-reductase inhibitor.
23. The method as defined in claim 22 wherein the
5-.alpha.-reductase inhibitor is finasteride.
Description
REFERENCE TO OTHER APPLICATIONS
[0001] This is a continuation-in-part of U.S. application Ser. No.
09/447,002, filed Nov. 22, 1999, which is a continuation of U.S.
application Ser. No. 09/119,884, filed Jul. 21, 1998, now U.S. Pat.
No. 6,013,668, which takes priority from Provisional Application
No. 60/055,568, Aug. 13, 1997, and Provisional Application No.
60/071,364, Jan. 15, 1998.
FIELD OF THE INVENTION
[0002] The present invention relates to a method for inhibiting
hair loss and/or promoting hair growth in chemotherapy patients
employing the (R)-enantiomer of 4-[
[(cyanoimino)[(1,2,2-trimethylpropyl)amino]methyl]a- mino]
benzonitrile or pharmaceutical compositions containing same.
BACKGROUND OF THE INVENTION
[0003] Potassium channel openers such as minoxidil (Upjohn),
pinacidil (Lilly) and diazoxide (Shiseido and Schering-Plough) are
known for their hair growth stimulating activity. Thus, U.S. Pat.
Nos. 4,596,812 and 4,139,619 disclose use of minoxidil in the
treatment of male pattern baldness, alopecia areata and balding in
females. U.S. Pat. No. 4,057,636 discloses pinacidil. DE 3,827,467A
discloses combinations of minoxidil and hydrocortisone or
retinoids.
[0004] U.S. Pat. No. 5,011,837 to Atwal et al discloses aryl
cyanoguanidines which possess potassium channel activating activity
and are useful therapy for hypertension and other cardiovascular
disorders, for various central nervous system disorders, kidney and
urinary problems as well as for the promotion of hair growth, for
example in the treatment of male pattern baldness (alopecia). These
aryl cyanoguanidines have the structure 1
[0005] and its possible tautomers 2
[0006] and 3
[0007] including pharmaceutically acceptable salts, wherein
[0008] R.sub.1 is alkyl, alkenyl, alkynyl, haloalkyl,
[0009] cycloalkyl, aryl, arylalkyl or cycloalkylalkyl;
[0010] R.sub.2 is 4
[0011] R.sub.3 and R.sub.4 are each independently selected form
-R.sub.2, hydrogen, alkyl, alkenyl, alkynyl, haloalkyl, halo,
alkoxy, --NHalkyl, --N-(alkyl).sub.2, --S-alkyl, --O-aryl-alkyl,
--S-arylalkyl or --S-aryl, --O-aryl, --NHaryl-alkyl, or R.sub.2 and
R.sub.3 taken together are a group which form a ring with the two
carbon atoms to which they are attached, which group is selected
from 5
[0012] wherein
[0013] m=1 or 2,
[0014] n=3-5,
[0015] p=2-4,
[0016] X is 0, NR.sub.5, CH.sub.2; and
[0017] R.sub.5 is hydrogen or R.sub.1.
[0018] Example 1 of U.S. Pat. No. 5,011,837 discloses the
preparation of 4-[[(cyanoimino)-[(1,2,2-trimethylpropyl)amino]
benzonitrile 6
[0019] in the form of its racemic mixture.
[0020] U.S. Pat. No. 6,013,668 discloses a method for promoting
hair growth in humans employing the (R)-enantiomer of
4-[[(cyanoimino)[(1,2,2-- trimethylpropyl)-amino]
methyl]amino]benzonitrile.
[0021] PCT Application WO 92/02225 discloses a combination of a
potassium channel opener and a 5-.alpha.-reductase inhibitor for
promoting hair growth.
[0022] PCT Application WO 92/09259A discloses use of an androgen
blocker and a potassium channel activator for stimulation of hair
growth.
[0023] PCT Application WO 96/29988 discloses a topical formulation
containing minoxide or minoxidil in combination with a testosterone
5-.alpha. reductase inhibitor.
[0024] PCT Application WO 94/18936 discloses a method for promoting
hair growth employing a vasodilator such as minoxidil in
combination with estradiol and/or a 5-.alpha.-reductase
inhibitor.
[0025] The use of minoxidil in cancer patients to decrease the
duration of baldness caused by chemotherapy is disclosed by Duvic,
M. et al, "A randomized trial of minoxidil in chemotherapy-induced
alopecia", J. Am. Acad Dermalol 1996; 35:74-8. Duvic et al disclose
that in patients treated with fluorouracil, doxorubicin and
cyclophosphamide a 2% topical solution of minoxdil administered
during chemotherapy and 4 weeks thereafter, did not prevent
alopecia but did decrease period of baldness.
[0026] Rodriguez, R. et al "Minoxidil (Mx) as a prophylaxis of
doxorubicin-induced alopecia", Annals of Oncology 5:769-770, 1994
discloses that a 2% topical solution of minoxidil was not effective
in preventing doxorubicin-induced alopecia.
[0027] Hussein, A. M., "Protection Against Cytosine
Arabinoside-Induced Alopecia By Minoxidil In A Rat Animal Model",
Int. J. Dermatol. Vol. 34(7); 470-3, 1995 discloses that minoxidil,
when injected locally, offered good local prevention against
1-B--D-arabinofurano-sylcytosine but not cyclophosphamide-induced
alopecia.
DESCRIPTION OF THE INVENTION
[0028] In accordance with the present invention, a method is
provided for preventing or inhibiting chemotherapy-induced or
radiation therapy-induced hair loss wherein a therapeutically
effective amount of the (R)-enantiomer of
4-[[(cyanoimino)[(1,2,2-trimethylpropyl)-amino]
methyl]amino]benzonitrile, (hereinafter "the (R)-enantiomer") is
administered to a human or other mammal.
[0029] In addition, in accordance with the present invention, a
method is provided for promoting hair growth in a patient
undergoing chemotherapy or radiation and/or having
chemotherapy-induced hair loss or radiation-induced hair loss,
wherein a therapeutically effective amount of the (R)-enantiomer is
administered to the patient.
[0030] In carrying out the above methods, the (R)-enantiomer will
be administered to the patient prior to and/or simultaneously with
and/or subsequent to chemotherapy and/or radiation therapy.
[0031] The above (R)-enantiomer of the invention has the structure
I 7
[0032] The (R)-enantiomer will preferably be in substantially pure
form, that is, will be at least 99% pure (R)-enantiomer and will at
most contain 1% (S)-enantiomer.
[0033] The method of the present invention also includes the use of
pharmaceutical compositions containing the (R)-enantiomer and a
pharmaceutically acceptable carrier therefor.
[0034] The (R)-enantiomer may be prepared as described in U.S. Pat.
No. 6,013,668 which is incorporated herein by reference.
[0035] The (R)-enantiomer may be administered by itself or may be
administered prior to, simultaneous with or after the
antineoplastic agent used in chemotherapy, or prior to
simultaneously with or after radiation therapy. In a preferred
embodiment of the present invention, the (R)-enantiomer is
administered prior to the antineoplastic agent or radiation
therapy.
[0036] As used herein, the term "simultaneous" means that the
antineoplastic agent or radiation therapy and the (R)-enantiomer
are administered within 24 hours, preferably 12 hours, more
preferably 6 hours, and most preferably 3 hours, of each other.
[0037] The chemotherapeutic agent which may be employed with the
(R)-enantiomer may include any of the antineoplastic agents listed
in the Physician's Desk Reference.
[0038] As used herein, the phrase "radiation therapy" includes, but
is not limited to, x-rays or gamma rays which are delivered from
either an externally applied source such as a beam or by
implantation of small radioactive sources.
[0039] As used herein, the phrase "antineoplastic agent" refers to
compounds which prevent cancer cells from multiplying. In general,
the antineoplastic agents of this invention prevent cancer cells
from multiplying by: (1) interfering with the cell's ability to
replicate DNA, or (2) inducing apoptosis in the cancerous
cells.
[0040] Examples of antineoplastic agents which are suitable for use
in the methods of this invention include, but are not limited to,
microtuble-stabilizing agents such as the taxanes, for example,
paclitaxel (also known as Taxol.RTM.), docetaxel (also known as
Taxotere.RTM.), 7-O-methylthio-methylpaclitaxel (disclosed in U.S.
Pat. No. 5,646,176),
3'-tert-butyl-3'-N-tert-butyloxycarbonyl-4-deacetyl-3'-de-
phenyl-3'-N-debenzoyl-4-O-methoxycarbonyl-paclitaxel (disclosed in
U.S. Ser. No. 60/179,965 filed on Feb. 3, 2000, and example 17
herein), C-4 methyl carbonate paclitaxel (disclosed in WO
94/14787), the epothilones, such as epothilone A, epothilone B,
epothilone C, epothilone D, desoxyepothilone A, desoxyepothilone B,
[1S-[ 1R*,3R*(E),7R*,10S*,11R*,12-
R*,16S*]]-7,11-dihydroxy-8,8,10,12,16-pentamethyl-3-[1-methyl-2-(2-methyl--
4-thiazolyl)ethenyl]-4-aza-17-oxabicyclo[14.1.0]hepta-decane-5,9-dione
(disclosed in WO 99/02514), [1S-[
1R*,3R*(E),7R*,10S*,11R*,12R*,16S*]]-3--
[2-[2-(aminomethyl)-4-thiazolyl]-1-methylethenyl]-7,11-di-hydroxy-8,8,10,1-
2,16-pentamethyl-4,17-dioxabi-cyclo[14.1.0]-heptadecane-5,9-dione
(disclosed in U.S. Ser. No. 09/506,481 filed on Feb. 17, 2000, and
examples 7 and 8 herein), and derivatives thereof;
microtuble-disruptor agents; alkylating agents; anti-metabolites;
epidophyllotoxin; an antineoplastic enzyme; a topoisomerase
inhibitor; procarbazine; mitoxantrone; platinum coordination
complexes; biological response modifiers; growth inhibitors;
hormonal/antihormonal therapeutic agents; and haematopoietic growth
factors.
[0041] Other classes of antineoplastic agents suitable for use in
the method of the present invention include, but are not limited
to, the anthracycline family of drugs, the vinca drugs, the
mitomycins, the bleomycins, the cytotoxic nucleosides,
discodermolide, the pteridine family of drugs, diynenes, aromatase
inhibitors, and the podophyllotoxins. Particularly useful members
of those classes not previously mentioned include, for example,
doxorubicin, carminomycin, daunorubicin, aminopterin, methotrexate,
methopterin, dichloro-methotrexate, mitomycin C, porfiromycin,
5-fluorouracil, 6-mercaptopurine, gemcitabine, cytosine
arabinoside, podophyllotoxin or podophyllotoxin derivatives such as
etoposide, etoposide phosphate or teniposide, melphalan,
vinblastine, vincristine, leurosidine, vindesine, leurosine, and
the like. Other useful antineoplastic agents include estramustine,
cisplatin, carboplatin, cyclophosphamide, bleomycin, tamoxifen,
ifosfamide, melphalan, hexamethyl melamine, thiotepa, cytarabin,
idatrexate, trimetrexate, dacarbazine, L-asparaginase,
camptothecin, CPT-11, topotecan, ara-C, bicalutamide, flutamide,
leuprolide, pyridobenzoindole derivatives, interferons, and
interleukins.
[0042] In carrying out the method of the invention, the
(R)-enantiomer may be formulation with other hair growth promoting
compounds such as the potassium channel openers minoxidil (Upjohn)
and/or diazoxide (Shiseido and Schering-Plough), as well as
cromakalim and pinacidil; a 5-.alpha.-reductase inhibitor such as
finasteride (Merck's Proscar.RTM.), terazosin HCl (Abbott's
Hytrin.RTM.), or doxaosin mesylate (Pfizer's Cardura.RTM.); and/or
an androgen blocker such as 4-(5-methoxyheptyl)-hex-
ahydro-2(1H)-pentalenone as disclosed in PCT Application WO
92/09259A, vasoconstrictors such as betamethasone dipropionate,
corticosteroids such as hydrocortisone, and scopolamine, and
cyproterone acetate.
[0043] The (R)-enantiomer may be administered via topical, oral,
parenteral or rectal routes as described in U.S. Pat. No. 5,011,837
(incorporated herein by reference), with topical being preferred,
to humans or other mammals such as dogs and cats prior to,
simultaneous with and/or subsequent to chemotherapy and/or
radiation therapy. Thus, the (R)-enantiomer in suitable topical
formulations is applied to the skin region where hair growth is
desired and/or where hair loss is to be inhibited.
[0044] Typical topical formulations for use herein will include
conventional ointments, creams, lotions, waxes, gels, pastes,
jellies, sprays, aerosols and the like in aqueous or non-aqueous
formulations. Examples of suitable topical formulations are
disclosed in U.S. Pat. Nos. 4,139,619 and 4,596,812 which are
incorporated herein by reference.
[0045] The (R)-enantiomer will be used in an effective amount, that
is, in an amount sufficient to inhibit hair loss during
chemotherapy and/or radiation therapy and/or promote hair growth
during and/or subsequent to chemotherapy and/or radiation therapy,
such that hair growth is increased or produced. A typical topical
composition will include from about 0.01 to about 15% by weight,
preferably from about 0.1 to about 10% by weight of the
composition.
[0046] The topical formulations containing the (R)-enantiomer of
the invention can be applied to the area to be treated such as the
scalp in humans, by spraying, dabbing or swabbing to deliver the
enantiomer to the region of the hair follicle. The formulations
will be applied to the area of treatment on a routine basis prior
to, during and subsequent to chemotherapy and/or radiation therapy,
at least once daily, and preferably two or more times daily.
[0047] The accompanying Figure is a graph showing the effect of a
once daily application of each of the (R)-and (S)-enantiomers
described herein on hair growth in male C3H mice.
[0048] The following Example describes the preparation of the
(R)-enantiomer and the (S)-enantiomer.
EXAMPLE 1
[0049] (R)-4-[[(Cyanoimino)[(1,2,2-trimethylpropyl)amino]-methyl]
amino]benzonitrile 8
[0050] A. (R)-1,2,2-Trimethylpropyl amine 9
[0051] The title compound was prepared according to the procedure
described by Manley and Quast (J. Med. Chem. 1992, 35, 2327-2340)
with some modification. A mixture of pinacolone (29 g, 290 mmol),
(R)-.alpha.-methylbenzyl amine (17.6 g, 145 mmol) and
p-toluenesulfonic acid monohydrate (300 mg) in toluene (150 mL) was
refluxed using a Dean-Stark trap (to remove water from the reaction
mixture) for 3 days. The solvent was evaporated and the residue was
distilled at ca. 120-2.degree. C. (9 mm) to give 21 g (71% yield)
of 10
[0052] as a colorless oil. This material was dissolved in anhydrous
THF (210 mL) and treated at 0-2.degree. C. with borane-THF complex
(1M, 206 mL, 206 mmol). The mixture was allowed to come to room
temperature, stirred for 5h and concentrated in vacuo. To the
resulting oily residue was carefully added ethanol (300 mL), and
the mixture was refluxed for 1h and concentrated again in vacuo.
The residue was chromatographed over basic alumina (activity grade
1/hexane) giving colorless oil. Proton NMR and HPLC (YMC C18 S3
4.6X50 mm column/water-MeOH--H.sub.3PO.sub.4 90:10:0.2 to 10:90:0.2
gradient) indicated that this material was contaminated with ca.
10% of the (S,R)-diastereomer. Therefore, this mixture was
resubjected to flash chromatography (silica
gel/hexane-EtoAc-triethylamine 95:5:0.1) to afford 11
[0053] (11.45 g, 55.8 mmol, 54% yield). The above compound (11.45
g) and 10% palladium on carbon (1.5 g) were taken in EtOH (230 mL)
and stirred under hydrogen for 12 hours. The mixture was filtered
and the filtrate (ca. 230 mL) containing the title product was used
as such for the next step as a ca. 0.24 M solution in ethanol
(assumed 100% yield).
[0054] B. N-Cyano-N'-(4-cyanophenyl)thiourea, monosodium salt
12
[0055] The title compound was prepared according to Example 1 Part
A of U.S. Pat. No. 5,011,837.
[0056] C. (R)-4-[[(Cyanoimino)[(1,2,2-trimethyl-propyl)amino]
methyl]amino]benzonitrile 13
[0057] To a solution of Part B compound (6.0 g, 26.8 mmol) in DMF
(150 mL) was sequentially added the solution of Part A compound
(ca. 0.24 M in EtOH, 112 mL, 26.8 mmol) and
1-(3-dimethylamino-propyl)-3-ethylcarbodiimi- de hydrochloride
(WSC) (6.0 g, 31.3 mmol). The mixture was stirred at room
temperature for 3 hours, diluted with ethyl acetate and
sequentially washed with 1N HC1, water and brine. The organic layer
was dried over magnesium sulfate, concentrated and the crude
product was purified by flash chromatography on silica gel
(hexanes-ethyl acetate-triethylamine 75:25:0.2) to afford a
colorless foam. This material was recrystallized from isopropanol
to give the title compound as a white solid (4.15 g, 57.6%), mp
159-60.degree. C.; [.alpha.].sub.D-180.degree. C.=1, MeOH;
enantiomeric purity determined by chiral HPLC=99% (ChiralPak AD
column/hexane-isopropanol-triethylamine 80:20:0.2); MS: 270
(M+H).sup.+; .sup.1H NMR (CDCl.sub.3) .delta. 8.65 (br s, 1H), 7.69
(d, 2H, J=8.79 Hz), 7.37 (d, 2H, J=8.79 Hz), 4.93 (br d, 1H), 3.83
(m, 1H), 1.10 (d, 1H, J=6.45 Hz), 0.90 (s, 9H).
[0058] Elemental analysis: calculated for
C.sub.15H.sub.19N.sub.5:
[0059] C, 66.89; H, 7.11; N, 26.00
[0060] Found: C, 66.71; H, 7.14; N, 25.98.
EXAMPLE 2
[0061] (S)-4-[[(Cyanoimino)[(1,2,2-trimethylpropyl)amino]-methyl]
amino]benzonitrile 14
[0062] The title compound was prepared from Part B compound of
Example 1 and (S)-1,2,2-trimethylpropyl amine (prepared according
to Manley and Quast, J. Med. Chem., 1992, 35, 2327-2340) by the
same procedure as described in Example 1, Part C. The product was
obtained as a colorless solid, mp 158-59.degree. C.;
[a].sub.D+189.degree. C.=1, MeOH; enantiomeric purity determined by
chiral HPLC=99.4% (ChiralPak AD
column/hexane-isopropanol-triethylamine 80:20:0.2); MS: 270
(M+H).sup.+; .sup.1H NMR (CDCl.sub.3) .delta. 8.43 (br s, 1H), 7.69
(d, 2H, J=8.79 Hz), 7.37 (d, 2H, J=8.79 Hz), 4.93 (br d, 1H), 3.83
(m, 1H), 1.10 (d, 1H, J=6.45 Hz), 0.90 (s, 9H).
EXAMPLE 3
[0063] Comparison of Example 1-(R)-Enantiomer and Example
2-(S)-Enantiomer Re Hair Growth in an Animal Model
[0064] The objective of the following described experiment was to
compare and evaluate the in vivo effect of the Example
1-(R)-enantiomer and the Example 2-(S)-enantiomer on hair growth in
an animal model. The two enantiomers were compared topically for
hair growth in C3H mice.
[0065] Animal Model
[0066] The C3H mouse is a useful model for studying hair growth.
Its usefulness rests with the fact that skin pigmentation of this
animal is provided by the melanocytes of the hair follicle and not
the epidermis. In the telogen or the resting phase of the hair
follicle, the skin is pink. In the earliest phase of anagen or the
growth phase, there is sudden graying of the skin and as the anagen
phase progresses the skin becomes darker in color. In this study,
visual observation was used as an in vivo assay of anagen
induction. Furthermore as anagen develops, the skin thickness
increases from a thin telogen skin to a measurably thickened anagen
skin. Thus, recording the skin color and microscopic thickness of
skin from these mice offers a sensitive, quantifiable and
convenient method of assessing the phases of hair growth.
[0067] Groups of 20, six to seven week old male C3H mice with hair
follicles in the resting phase of hair growth were used. At this
stage in their life, the hair follicles remain in the telogen phase
for up to 30 days or longer. This provides an adequate window of
time to screen drugs. Compounds that improve hair growth stimulate
the hair follicles from the telogen to the anagen phase. This
stimulation is manifested by the shortening of the telogen phase of
the hair follicle cycle.
[0068] Animals were anesthetized with ketamine/rompun (100 mg/Kg
and 12 mg/Kg) IP and the hair over a defined dorsal area were
closely clipped.
[0069] Animals with pink skin were treated topically 1.times.
daily, 5 days per week with 50 microliters of a 2% solution of
Example 1-(R)-enantiomer and a 2% solution of Example
1-(S)-enantiomer or vehicle by itself, applied to the dorsal area.
The vehicle employed was ethanol/propylene glycol/water, 60/30/10.
Treatment was continued for at least 4-5 weeks.
[0070] Animals were observed daily for side effects and changes to
the test sites. All observations were documented. Test sites were
graded weekly for changes in skin color and hair growth. In this
study drug effects were evaluated using the visual observation of
skin changing from pink to gray and resulting in hair growth.
[0071] Results
[0072] The percent of animals that induced hair follicle
stimulation during the treatment period is illustrated in the
accompanying Figure below. The most significant observation made
between the two enantiomers is the difference in the time of onset
of follicle stimulation. The time of onset for the Example
1-(R)-enantiomer was day 7 compared to day 11 for Example
2-(S)-enantiomer. The time of onset for the vehicle control was day
28. By day 11 of treatment the Example 1-(R)-enantiomer caused hair
follicle stimulation in 40% of the test mice compared to only 5%
with Example 2-(S)-enantiomer. By day 14, 50% of the animals
treated with Example 1-(R)-enantiomer showed hair follicle
stimulation compared to 25% for Example 2-(S)-enantiomer. By day
28, 85% of the animals treated with the Example 1-(R)-enantiomer
showed hair follicle stimulation as compared to 65% treated with
Example 2-(S)-enantiomer. Thus throughout the treatment period, the
group treated with Example 1-(R)-enantiomer showed a higher
incidence of hair follicle stimulation as compared to the group
treated with Example 2-(S)-enantiomer.
[0073] The attached Figure shows the effect of 1.times. daily
topical application of Example 1-(R)-enantiomer and Example
2-(S)-enantiomer.
[0074] In conclusion, these results in the C3H mice indicate that
there is a remarkable difference between the Example
1-(R)-enantiomer and the Example 2-(S)enantiomer in their effect on
hair follicle stimulation; in particular the (R)-enantiomer has a
faster onset of action compared to the corresponding
(S)-enantiomer.
[0075] These results are indeed surprising and unexpected
especially in view of the vasorelaxant potencies of each of these
enantiomers, which is generally recognized as an indication of hair
growth promoting properties (Side Effects of Vasodilator Therapy,
W. A. Pettinger et al, Hypertension, 1988, Vol. 11, II-34 to II-36,
and Minoxidil Stimulates Cutaneous Blood Flow in Human Balding
Scalps: Pharmacodynamics measured by laser Doppler velocimetry and
photopulse plethysmography. R. C. Wester et al, J. Invest.
Dermatol., 184, Vol. 82, 515-517).
[0076] Thus, while the IC.sub.50 for vasorelaxant potency of the
(R)-enantiomer is 47.+-.17 nM versus 157.+-.35 nM for the
(S)-enantiomer, as seen above, the hair growth promoting ability of
the (R)-enantiomer for producing hair growth within 11 days of
treatment is 8 times greater than the corresponding
(S)-enantiomer.
* * * * *