Small molecule compound and synthesizing method and uses thereof

Abstract

Provided is a small molecule compound as represented by structural formula (I). The product of the present invention in various concentrations and dosages can achieve an obvious change in the growth period of hairs, promoting the growth of the hairs, thus exhibiting an obvious effect of promoting hair growth. In addition, changes in the weight of a mouse in each group are slow, indicating that the test compound does not cause weight loss in an animal.

Claims

1. A small molecule compound, characterized in that, it is represented by the following structural formula: ##STR00047## R.sub.1 is selected from hydrogen, halogen, C.sub.1-C.sub.6alkyl or optionally substituted with 1-3 substituents, cyano, isocyanate, amide, iso-sulfonamide (SO.sub.2NHR), iso-sulfinyl (SONH), sulfonamide (NHSO.sub.2R), sulfonamido (NHSOR), S-alkyl, S-aryl, S-heteroaryl (SHet); wherein the substituents are selected from halogen, aryl, substituted aryl, heteroaryl, O-alkyl, O-aryl, O-heteroaryl, N-alkyl, N-aryl, N-heteroaryl; R.sub.2 is selected from Hydrogen, halogen, C.sub.1-C.sub.6 alkyl or substituted alkyl, alkoxy, aryl, substituted aryl, benzyl, substituted benzyl; R.sub.3 is selected from Hydrogen, halogen, C.sub.1-C.sub.6 alkyl or substituted alkyl, alkoxy, aryl, substituted aryl, benzyl, substituted benzyl; R.sub.4 is selected from hydrogen, C.sub.1-C.sub.6 alkyl or substituted alkyl; R.sub.5 is selected from hydrogen, C.sub.1-C.sub.6 alkyl or substituted alkyl; X is selected from alkylene or substituted alkylene, amino or substituted amino; Y is selected from alkylene or substituted alkylene, oxygen, sulfur, amino, carbonyl, sulfoxide, sulfone.

2. The small molecule compound of claim 1, characterized in that, it is represented by the following structural formula: ##STR00048## wherein R is halogen, hydroxyl, hydrogen, or mono- or polysubstituted alkylsulphonyl, alkyl, methoxy, nitro, amino, carboxyl, ester, aryl, or benzyl.

3. The small molecule compound of claim 1, characterized in that, it is represented by the following structural formula: ##STR00049## wherein R is halogen, hydroxyl, hydrogen, or mono- or polysubstituted alkylsulphonyl, alkyl, methoxy, nitro, amino, carboxyl, ester, aryl, benzyl, O-alkyl, O-aryl, O-heteroaryl, N-alkyl, N-aryl, or N-heteroaryl; X is CH or nitrogen; Z.sub.1, Z.sub.2, Z.sub.3, Z.sub.4, Z.sub.5 are carbon, oxygen, sulfur, nitrogen, carbonyl; and X, Z.sub.1, Z.sub.2, Z.sub.3, Z.sub.4 and Z.sub.5 form aromatic or nonaromatic lactam.

4. The small molecule compound of claim 1, characterized in that, it is represented by the following structural formula: ##STR00050## wherein R is halogen, hydroxyl, hydrogen, mono- or polysubstituted alkylsulphonyl, alkyl, methoxy, nitro, amino, carboxyl, ester, aryl, benzyl, O-alkyl, O-aryl, O-heteroaryl, N-alkyl, N-aryl, N-heteroaryl; X is CH or nitrogen; Z.sub.1, Z.sub.2, Z.sub.3, Z.sub.4 are carbon, oxygen, sulfur, nitrogen, carbonyl, and X, Z.sub.1, Z.sub.2, Z.sub.3, and Z.sub.4 form aromatic or nonaromatic lactam.

5. The small molecule compound of claim 1, characterized in that, it is N1-(3,5-dichloro-4-(3-(4-fluorobenzyl)-4-hydroxyphenoxy)phenyl)-N2-oxalyl hydroxylamine; or N1-(3,5-dichloro-4-(3-(4-methylsulfonylbenzyl)-4-hydroxyphenoxy)phenyl)-N2-oxalyl hydroxylamine; or N1-(3,5-dichloro-4-(3-(4-methylbenzyl)-4-hydroxyphenoxy)phenyl)-N2-oxalyl hydroxylamine; or N1-(3,5-dichloro-4-(3-(3, 4-dichlorobenzyl)-4-hydroxyphenoxy)phenyl)-N2-oxalyl hydroxylamine; or N1-(3,5-dichloro-4-(3-(3-fluorobenzyl)-4-hydroxyphenoxy)phenyl)-N2-oxalyl hydroxylamine; or N1-(3,5-dichloro-4-(3-(2-fluorobenzyl)-4-hydroxyphenoxy)phenyl)-N2-oxalyl hydroxylamine; or N1-(3,5-dichloro-4-4-hydroxy-3-(morpholinomethyl)phenoxy)phenyl)-N2-oxalyl hydroxylamine; or N1-(3,5-dichloro-4-(4-hydroxy-3-(4-hydroxypiperidin-1-yl)phenoxy)phenyl)-N2-oxalyl hydroxylamine; or N1-(3,5-dichloro-4-(4-hydroxy-3-(4-methylpiperazin-1-yl)phenoxy)phenyl)-N2-oxalyl hydroxylamine; or N1-(3,5-dichloro-4-(4-hydroxy-3-(4-methylpiperazin-1-methyl)phenoxy)phenyl)-N2-oxalyl hydroxylamine; or N1-(3,5-dichloro-4-(4-hydroxy-3-(pyrrolidin-1-methyl)phenoxy)phenyl)-N2-oxalyl hydroxylamine; or N1-(4-(3-((4-benzylpiperidin-1-methyl)-4-hydroxyphenoxy)-3,5-dichlorophenyl)-N2-oxalyl hydroxylamine; or N1-(3,5-dichloro-4-(4-hydroxy-3-((4-hydroxy-4-phenylpiperidin-1-methyl)phenoxy)phenyl)-N2-oxalyl hydroxylamine; or N1-(3,5-dichloro-4-(4-hydroxy-3-(isoindolin-2-methyl)phenoxy)phenyl)-N2-oxalyl hydroxylamine; or N1-(3,5-dichloro-4-(3-(3,4-dihydroisoquinolin-2(H)-methyl)-4-hydroxyphenoxy)phenyl)-N2-oxalyl hydroxylamine; or N1-(3,5-dichloro-4-(4-hydroxy-3-(3-piperazinone-1-methyl)phenoxy)phenyl)-N2-oxalyl hydroxylamine.

6. A process for synthesizing a small molecule compound, characterized in that: step 1: 4-methoxyphenol is reacted with 1,2,3-trichloro-5-nitrobenzene to form 1,3-dichloro-2-(4-methoxyphenoxy)-5-nitrobenzene; step 2: 1,3-dichloro-2-(4-methoxyphenoxy)-5-nitrobenzene is reacted with benzoic acid derivatives to form (5-(2,6-dichloro-4-nitrophenoxy)-2-methoxyphenyl) (substituted phenyl) methyl ketone); step 3: 1,3-dichloro-2-(3-(substituted benzyl)-4-methoxyphenoxy)-5-nitrobenzene is obtained from reduction of carbonyl of (5-(2,6-dichloro-4-nitrophenoxy)-2-methoxyphenyl)(substituted phenyl) methyl ketone to methylene; step 4: 3,5-dichloro-4-(3-(substituted benzyl)-4-methoxyphenoxy)-aniline is obtained from reduction of nitro of 1,3-dichloro-2-(3-(substituted benzyl)-4-methoxyphenoxy)-5-nitrobenzene to amino; Step 5: 3,5-dichloro-4-(3-(substituted benzyl)-4-methoxyphenoxy)-aniline is reacted with ethyl oxaloyl monochloride to form 2-((3,5-dichloro-4-(3-(substituted benzyl)-4-methoxyphenoxy)phenyl)amino)-oxoacetate; Step 6: 2-((3,5-dichloro-4-(3-(substituted benzyl)-4-methoxyphenoxy) phenyl)amino)-oxoacetate is reacted with hydroxylamine hydrochloride to form N1-(3,5-dichloro-4-(3-(substituted benzyl)-4-methoxyphenoxy)phenyl)-N2-oxalyl hydroxylamine; Step 7: after demethylation of methoxyl of N1-(3,5-dichloro-4-(3-(substituted benzyl)-4-methoxyphenoxy)phenyl)-N2-oxalyl hydroxylamine, a target small molecule compound can be obtained.

7. The process for synthesizing a small molecule compound of claim 6, characterized in that: the derivatives of benzoic acid can be replaced with the compound having the following structure: ##STR00051## wherein R can be C.sub.1-C.sub.6 alkyl or optionally substituted with 1-3 substituents; the substituents are selected from halogen, aryl, substituted aryl, heteroaryl, O-alkyl, O-aryl, O-heteroaryl, N-alkyl, N-aryl, N-heteroaryl, S-alkyl, S-aryl and S-heteroaryl.

8. The process for synthesizing a small molecule compound of claim 6, characterized in that: in step 1, the molar ratio of 4-methoxyphenol to 1,2,3-trichloro-5-nitrobenzene is 1:1-2; in step 2, the molar ratio of 1,3-dichloro-2-(4-methoxyphenoxy)-5-nitrobenzene to benzoic acid derivative is 1:1.25-2; in step 4, the molar ratio of 1,3-dichloro-2-(3-(substituted benzyl)-4-methoxyphenoxy)-5-nitrobenzene to reducing agent is 1:10-20; in step 5, the molar ratio of 3,5-dichloro-4-(3-(substituted benzyl)-4-methoxyphenoxy)aniline to ethyl oxalyl monochloride is 1:1.5-3; in step 6, the molar ratio of 2-((3,5-dichloro-4-(3-(substituted benzyl)-4-methoxyphenoxy)phenyl)amino)-oxoacetate to hydroxylamine hydrochloride is 1:5-10.

9. The process for synthesizing a small molecule compound of claim 6, characterized in that: in step 1, the reaction is carried out under strong basic condition, the strong bases can be selected from KNH.sub.2, NaNH.sub.2, NaCN, KCN, butyl lithium, lithium diisopropylamine, benzyl lithium, Grignard reagents, lithium alkylcuprate, sodium methoxide, sodium ethoxide, potassium ethoxide, NaOtBu, NaOH or KOH, and the reaction temperature is 120-160 C.; in step 2, reaction is carried out in Eaton's Reagent, the reaction temperature is 80-130 C.

10. The process for synthesizing a small molecule compound of claim 6, characterized in that: in step 3, trifluoroacetic acid and triethyl silane are used to reduce the carbonyl group to methylene group, wherein the molar ratio of the reactants (5-(2,6-dichloro-4-nitrophenoxy)-2-methoxyphenyl)(substituted phenyl) methyl ketone:trifluoroacetic acid:triethyl silane 1:4-6:3-5.

11. The process for synthesizing a small molecule compound of claim 6, characterized in that: demethylation of step 7 can be carried out using aluminum trichloride (AlCl3), zinc chloride (ZnCl2) or boron tribromide (BBr3).

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) FIG. 1(a), Photograph of mice with hair growth in the TDM-001-1 group;

(2) FIG. 1(b), Photograph of mice with hair growth in the TDM-001-2 group;

(3) FIG. 1 (c), Photograph of mice with hair growth in the TDM-001-3 group;

(4) FIG. 1 (d), Photograph of mice with hair growth in the TDM-001-4 group;

(5) FIG. 2. Experimental results of TMI-105795;

(6) FIG. 3. Experimental results of TMI-105902;

(7) FIG. 4. Experimental results of TMI-105903;

(8) FIG. 5. Experimental results of TMI-105905;

(9) FIG. 6. Experimental results of TMI-105965;

(10) FIG. 7. Experimental results of TMI-105966;

(11) FIG. 8. Experimental results of TMI-105906;

(12) FIG. 9. Experimental results of TMI-105956;

(13) FIG. 10. Experimental results of TMI-105957;

(14) FIG. 11. Experimental results of TMI-105958;

(15) FIG. 12. Experimental results of TMI-105959;

(16) FIG. 13. Experimental results of TMI-105960;

(17) FIG. 14. Experimental results of TMI-105961;

(18) FIG. 15. Experimental results of TMI-105962;

(19) FIG. 16. Experimental results of TMI-105963;

(20) FIG. 17. Diagram of body weight changes of mice during administration

(21) FIG. 18. Diagram of scoring on hair growth during administration

(22) FIG. 19. Diagram of scoring on scaling during administration;

(23) FIG. 20. Diagram of scoring on erythema during administration.

EXAMPLES

Example 1. Product Number: TMI-105795

Name

N1-(3,5-dichloro-4-(3-(4-fluorobenzyl)-4-hydroxyphenoxy)phenyl)-N2-oxalyl hydroxylamine

(24) Structural Formula:

(25) ##STR00023##
Synthesizing Route:

(26) ##STR00024## ##STR00025##

Specific Synthesizing Method

Step 1: Synthesis of 1,3-dichloro-2-(4-methoxyphenoxy)-5-nitrobenzene

(27) ##STR00026##

(28) 4-methoxyphenol (5.5 g, 44 mmol) was dissolved in DMF (100 mL) and treated with NaH (2.64 g, 66 mmol). After stirring for 30 minutes, 1,2,3-trichloro-5-nitrobenzene (10 g, 44 mmol) was added. The reaction solution was heated to 120 C. for three hours. After completion of reaction, the mixture was cooled to room temperature and then condensed. The residue was quenched with water and extracted with ethyl acetate. The organic layer was dried and concentrated, the crude product was obtained; after column chromatography (0-10% ethyl acetate and petroleum ether), the product was obtained (5.0 g, 36%).

(29) In this step, different reaction conditions are used. For example, different reactant ratio (4-methoxyphenol:NaH:1,2,3-trichloro-5-nitrobenzene=1:2:1 or 1:1.8:1.5, etc.); solvents (chlorobenzene, dioxane, etc); bases (NaH, NaOH, NaOtBu, etc); reaction temperature (130 C., 160 C.); reaction time (1, 2, 4, 5, 6 hours, etc). The reaction yield varies from 31% to 68%.

Step 2: Synthesis of 5-(2,6-dichloro-4-nitrophenoxy)-2-methoxyphenyl)(4-fluorophenyl) methyl ketone

(30) ##STR00027##
1,3-Dichloro-2-(4-methoxypehnoxy)-5-nitrobenzene (5.0 g, 16 mmol), 4-fluorobenzoic acid (2.8 g, 20 mmol) was distributed in Eaton Reagent (30 mL). The reaction mixture was then heated to 80 C. for 4 hours. After cooling to room temperature, the mixture was quenched with sodium bicarbonate solution. The aqueous phase was extracted with ethyl acetate, dried anhydrously over Na.sub.2SO.sub.4 and concentrated. The residue was purified on column chromatographically (10% ethyl acetate and petroleum ether). The title compound was obtained (4.5 g, 64%).

(31) In this step, various reaction conditions were also tried, such as reactant ratio (1,3-dichloro-2-(4-methoxyphenoxy)-5-nitrobenzene:4-fluorobenzoic acid=1:1, 1:1.5, 1:2, etc.); reaction temperature (100 C., 110 C., 130 C., etc); reaction time (2, 5, 8 hours, etc). The reaction yield varies from 56% to 78%.

Step 3: Synthesis of 1,3-dichloro-2-(3-(4-fluorobenzyl)-4-methoxyphenoxy)-5-nitrobenzene

(32) ##STR00028##
5-(2,6-dichloro-4-nitrophenoxy)-2-methoxyphenyl)(4-fluorophenyl) methyl ketone (4.4 g, 10 mmol) was dissolved in DCM (50 mL) added with trifluoroacetic acid (6.0 g, 40 mmol) and triethyl silane (3.5 g, 30 mmol). The reaction solution was stirred for 3 hours at room temperature. The mixture was quenched with sodium bicarbonate solution. The aqueous phase was extracted with ethyl acetate, dried anhydrously over Na.sub.2SO.sub.4 and concentrated. The residue was purified on column chromatographically (10% ethyl acetate and petroleum ether). The title compound was obtained (3.5 g, 83%).

(33) In this step, various reaction conditions were also tried, such as reactant ratio (5-(2,6-dichloro-4-nitrophenoxy)-2-methoxyphenyl)(4-fluorophenyl) methyl ketone:trifluoroacetic acid:triethyl silane=1:4:3, 1:5:4, 1:6:5, etc); solvents (chloroform, chlorobenzene, toluene, etc.); reaction time (1, 2, 6 hours, etc.). The reaction yield varies from 80% to 96%.

Step 4: Synthesis of 3,5-dichloro-4-(3-(4-fluorobenzyl)-4-methoxyphenoxy)aniline

(34) ##STR00029##

(35) 1,3-dichloro-2-(3-(4-fluorobenzyl)-4-methoxyphenoxy)-5-nitrobenzene (3.5 g, 8.3 mmol) was dissolved in ethyl acetate (50 mL), followed by addition of Sn.sub.2Cl.sub.2 (18.7 g, 83 mmol). The reaction mixture was heated and refluxed for 3 hours and then cooled to room temperature. The mixture was adjusted to pH 8 using sodium bicarbonate solution and filtered and extracted. After concentration of filtered solution, the product was obtained (2.6 g, 81%).

(36) In this step, various reaction conditions were also tried, such as reactant ratio (1,3-dichloro-2-(3-(4-fluorobenzyl)-4-methoxyphenoxy)-5-nitrobenzene:Sn.sub.2Cl.sub.2=1:10, 1:15, 1:20 etc.); solvents (methyl acetate, acetone, benzene, etc); reducing agents (Zn/HCl, Pt/H.sub.2, Ni/H.sub.2, etc.); reaction time (1, 2, 6 hours, etc.). The reaction yield varies from 80% to 99%.

Step 5: Synthesis of 2-((3,5-dichloro-4-(3-(4-fluorobenzyl)-4-methoxyphenoxy)phenyl)amino)-oxoacetate (ethyl oxalyl)

(37) ##STR00030##
3,5-dichloro-4-(3-(4-fluorobenzyl)-4-methoxyphenoxy)aniline (200 mg, 0.51 mmol) was dissolved in CH.sub.2Cl.sub.2 and added with ethyl oxalyl monochloride (120 mg, 0.76 mmol) and triethylamine (100 mg, 1.02 mmol). After one hour of stirring, the reaction mixture was quenched with sodium bicarbonate solution. The aqueous phase was extracted with CH.sub.2Cl.sub.2 and the organic layer was washed with saturated brine and dried anhydrously over Na.sub.2SO.sub.4. After concentration, the product was obtained (250 mg, 100%).

(38) In this step, various reaction conditions were also tried, such as reactant ratio (the molar ratio of 3,5-dichloro-4-(3-(4-fluorobenzyl)-4-methoxyphenoxy)aniline:ethyl oxalyl monochloride=1:1.5, 1:2, 1:2.3, 1:3, etc, ethyl oxalyl monochloride:triethylamine=1:1.5, 1:2, 1:2.3, etc.); solvents (chloroform, ethyl acetate, chlorobenzene, etc.); reaction time (0.5, 2, 3 hours, etc). The reaction yield varies from 90%-100%.

Step 6: Synthesis of N1-(3,5-dichloro-4-(3-(4-fluorobenzyl)-4-methoxyphenoxy)phenyl)-N2-oxalyl hydroxylamine

(39) ##STR00031##
2-((3,5-dichloro-4-(3-(4-fluorobenzyl)-4-methoxyphenoxy)phenyl)amino)-oxoacetate (ethyl oxalyl) (50 mg, 0.10 mmol), hydroxylamine hydrochloride (35 mg, 0.5 mmol) and NaOH (20 mg, 0.5 mmol) were dissolved in Ethanol (EtOH) (10 mL). The reaction mixture was stirred at room temperature overnight. After removal of solvent, the residue was extracted with ethyl acetate. After drying and concentration, the title compound was obtained (50 mg, 100%).

(40) In this step, various reaction conditions were also tried, such as reactant ratio (the molar ratio of 2-((3,5-dichloro-4-(3-(4-fluorobenzyl)-4-methoxyphenoxy)phenyl)-amino)-oxoacetate (ethyl oxalyl): hydroxylamine hydrochloride:NaOH=1:4:4; 1:6:6; 1:8:8, 1:10:10, etc.); solvents (ethyl acetate, Methanol (MeOH), etc); reaction time (8, 10, 12, 14, 16,d 18 hours, etc). The reaction yield varies from 90% to 100%.

Step 7: Synthesis of N1-(3,5-dichloro-4-(3-(4-fluorobenzyl)-4-hydroxyphenoxy)phenyl)-N2-oxalyl hydroxylamine

(41) ##STR00032##
N1-(3,5-dichloro-4-(3-(4-fluorobenzyl)-4-methoxyphenoxy) phenyl)-N2-oxalyl hydroxylamine (50 mg, 0.1 mmol) was dissolved in CH.sub.2Cl.sub.2 and added with 2 drops of BBr.sub.3. The reaction mixture was stirred for 3 hours followed by MeOH quenching. The residues are dispersed in ethyl acetate and water, after concentration of ethyl acetate layer, n-hexane is added and the product is separated out (6 mg, 12%).

(42) In this step, various reaction conditions were also tried, such as BBr.sub.3 (1 mg, 5 mg, 10 mg, 15 mg, 20 mg, etc); solvents (CH.sub.2Cl.sub.2, CHCl.sub.3 and CCl.sub.4, etc); reaction time (1, 2, 5 hours, etc). The reaction yield varies from 4% to 30%.

(43) Spectrum Data of Product:

(44) H.sup.1-NMR (MeOD) 7.92 (s, 2H), 7.19 (dd, J=8.7, 5.5 Hz, 2H), 6.96 (t, J=8.9 Hz, 2H), 6.71 (d, J=9.1 Hz, 1H), 6.44-6.51 (m, 2H), 3.87 (s, 2H) Mass Spectrum: m/z 465.3[M+H].sup.+

(45) The following synthesizing methods used in Examples 2-6 are similar or close to that in Example 1, and they are not repeated herein accordingly.

Example 2 Product Number: TMI-105902

Name

N1-(3,5-dichloro-4-(3-(4-methylsulphonylbenzyl)-4-hydroxyphenoxy)phenyl)-N2-oxalyl hydroxylamine

(46) Structural Formula:

(47) ##STR00033##
Spectrum Data of Product

(48) .sup.1H NMR (MeOD) 7.92 (s, 2H), 7.83 (d, J=8.6 Hz, 2H), 7.47 (d, J=8.3 Hz, 2H), 6.73 (d, J=9.4 Hz, 1H), 6.49-6.57 (m, 2H), 4.02 (s, 2H), 3.10 (s, 3H)

(49) Mass Spectrum: m/z 523.6[MH].sup.

Example 3 Product Number: TMI-105903

Name

N1-(3,5-dichloro-4-(3-(4-methylbenzyl)-4-hydroxyphenoxy)phenyl)-N2-oxalyl hydroxylamine

(50) Structural Formula:

(51) ##STR00034##
Spectrum Data of Product:

(52) .sup.1H NMR (MeOD) 7.90 (s, 2H), 7.05 (s, 4H), 6.70 (d, J=8.6 Hz, 1H), 6.43-6.49 (m, 1H), 6.39 (d, J=3.0 Hz, 1H), 3.84 (s, 2H), 2.29 (s, 3H)

(53) Mass Spectrum: m/z 459.6[MH].sup.

Example 4 Product Number: TMI-105905

Name

N1-(3,5-dichloro-4-(3-(3,4-dichlorobenzyl)-4-hydroxyphenoxy)phenyl)-N2-oxalyl hydroxylamine

(54) Structural Formula:

(55) ##STR00035##
Spectrum Data of Product

(56) .sup.1H NMR (MeOD) 7.93 (s, 2H), 7.38 (d, J=8.1 Hz, 1H), 7.32 (d, J=1.9 Hz, 1H), 7.12 (dd, J=8.2, 2.0 Hz, 1H), 6.73 (d, J=8.6 Hz, 1H), 6.52-6.58 (m, 1H), 6.50 (d, J=3.0 Hz, 1H), 3.87 (s, 2H)

(57) Mass Spectrum: m/z 513.5[MH].sup.

Example 5 Product Number TMI-105965

Name

N1-(3,5-dichloro-4-(3-(3-fluorobenzyl)-4-methoxyphenoxy)phenyl)-N2-oxalyl hydroxylamine

(58) Structural Formula:

(59) ##STR00036##
Spectrum Data of Product:

(60) .sup.1H NMR (MeOD) 7.91 (s, 2H), 7.11-7.24 (m, 2H), 6.98-7.08 (m, 2H), 6.71 (d, J=8.6 Hz, 1H), 6.48 (dd, J=8.6, 3.2 Hz, 1H), 6.42 (d, J=3.0 Hz, 1H), 3.91 (s, 2H)

(61) Mass Spectrum: m/z 463.5[MH].sup.

Example 6 Product Number: TMI-105966

Name

N1-(3,5-dichloro-4-(3-(2-fluorobenzyl)-4-methoxyphenoxy)phenyl)-N2-oxalyl hydroxylamine

(62) Structural Formula:

(63) ##STR00037##
Spectrum Data of Product:

(64) .sup.1H NMR (MeOD) 7.91 (s, 2H), 7.12-7.24 (m, 2H), 7.00-7.10 (m, 2H), 6.71 (d, J=8.6 Hz, 1H), 6.48 (dd, J=8.7, 3.1 Hz, 1H), 6.42 (d, J=3.0 Hz, 1H), 3.91 (s, 2H) Mass Spectrum: m/z 463.5[MH].sup.

Example 7 Product Number: TMI-105906

Name

N1-(3,5-dichloro-4-(4-hydroxy-3-(morpholinomethyl)phenoxy)phenyl)-N2-oxalyl hydroxylamine

(65) Structural Formula:

(66) ##STR00038##
Spectrum Data of Product

(67) .sup.1H NMR (DMSO-d.sub.6) : 10.99 (br. s., 1H), 9.83 (br. s., 2H), 8.95 (s, 1H), 7.96-8.22 (m, 2H), 7.60-7.73 (m, 1H), 6.64-6.72 (m, 2H), 6.51-6.60 (m, 1H), 3.57 (br. s., 4H), 3.52 (s, 2H), 2.40 (br. s., 4H)

Example 8 Product Number: TMI-105956

Name

N1-(3,5-dichloro-4-(4-hydroxy-3-(4-hydropiperidine-1-yl)phenoxy)phenyl)-N2-oxalyl hydroxylamine

(68) Structural Formula:

(69) ##STR00039##
Spectrum Data of Product:

(70) .sup.1H NMR (DMSO-d.sub.6) : : 10.10 (br. s., 1H), 8.10 (s, 3H), 6.58-6.73 (m, 2H), 6.53 (d, J=8.3 Hz, 1H), 4.62 (br. s., 1H), 3.37 (br. s., 2H), 2.69 (br. s., 2H), 2.14 (br. s., 2H), 1.72 (d, J=9.4 Hz, 2H), 1.38 (d, J=9.1 Hz, 2H)

Example 9 Product Number TMI-105957

Name

N1-(3,5-dichloro-4-(4-hydroxy-3-(4-methylpiperazine-1-methyl)phenoxy)phenyl)-N2-oxalyl hydroxylamine

(71) Structural Formula:

(72) ##STR00040##
Spectrum Data of Product:

(73) .sup.1H NMR (DMSO-d.sub.6) : 11.01 (br. s., 1H), 8.06-8.14 (m, 2H), 6.60-6.70 (m, 2H), 6.54 (dd, J=8.7, 3.1 Hz, 1H), 3.34 (br. s., 2H), 2.42 (br. s., 4H), 2.18-2.38 (m, 4H), 2.11-2.18 (m, 3H)

Example 10 Product Number: TMI-105958

Name

N1-(3,5-dichloro-4-(4-hydroxy-3-(methylpyrrolidin-1-methyl)phenoxy)phenyl)-N2-oxalyl hydroxylamine

(74) Structural Formula:

(75) ##STR00041##
Spectrum Data of Product:

(76) .sup.1H NMR (DMSO-d.sub.6): : 10.90 (br.s., 1H), 10.77 (br.s., 1H), 8.11 (br.s., 2H), 6.87 (br.s., 1H), 6.64 (br.s., 2H), 6.53 (br.s., 2H), 3.45 (d, J=5.4 Hz, 2H), 1.72 (br.s., 4H), 1.06 (br.s., 2H)

Example 11 Product Number: TMI-105969

Name

N1-(4-(3-((4-benzylpiperidin-1-methyl)-4-hydroxyphenoxy)-3,5-dichlorophenyl)-N2-oxalyl hydroxylamine

(77) Structural Formula:

(78) ##STR00042##
Spectrum Data of Product:

(79) .sup.1H NMR (METHANOL-d.sub.4) : 8.40 (br. s., 1H), 7.99 (s, 2H), 7.51 (d, J=7.3 Hz, 2H), 7.38 (t, J=7.5 Hz, 2H), 7.30 (d, J=7.5 Hz, 1H), 6.87-6.98 (m, 2H), 6.82-6.87 (m, 1H), 4.34 (s, 2H), 3.38-3.52 (m, 4H), 2.28-2.41 (m, 2H), 1.96 (d, J=14.2 Hz, 3H), 1.27-1.39 (m, 2H)

Example 12 Product Number: TMI-105960

Name

N1-(3,5-dichloro-4-(4-hydroxy-3-((4-hydroxy-4-phenylpiperidin-1-methyl)phenoxy)phenyl)-N2-oxalyl hydroxylamine

(80) ##STR00043##
Spectrum Data of Product

(81) .sup.1H NMR (DMSO-d) : 10.63 (br. s., 1H), 8.10 (s, 2H), 7.22-7.32 (m, 2H), 7.10-7.22 (m, 3H), 6.57-6.68 (m, 2H), 6.51 (dd, J=8.6, 3.0 Hz, 1H), 3.35 (br. s., 2H), 2.82 (d, J=10.7 Hz, 2H), 1.96 (t. J=11.1 Hz, 2H), 1.43-1.68 (m, 2H), 1.09-1.26 (m, 2H)

Example 13 Product Number: TMI-105961

Name

N1-(3,5-dichloro-4-(4-hydroxy-3-(isoindolin-2-methyl)phenoxy)phenyl)-N2-oxalyl hydroxylamine

(82) Structural Formula:

(83) ##STR00044##
Spectrum Data of Product

(84) .sup.1H NMR (DMSO-d.sub.6) : 8.09 (s, 2H), 7.07-7.36 (m, 4H), 6.64-6.82 (m, 4H), 6.56 (dd, J=8.5, 3.4 Hz, 2H), 3.79-3.91 (m, 4H), 1.33 (d, J=4.8 Hz, 2H)

Example 14 Product Number: TMI-105962

Name

N1-(3,5-dichloro-4-(3-(3,4-dihydroisoquinolin-2(1H)-methyl)-4-hydroxyphenoxy)phenyl)-N2-oxalyl hydroxylamine

(85) Structural Formula:

(86) ##STR00045##
Spectrum Data of Product:

(87) .sup.1H NMR (DMSO-d.sub.6) : 10.03 (br. s., 1H), 8.05-8.16 (m, 1H), 7.62 (s, 1H), 7.08-7.17 (m, 2H), 7.04 (d, J=5.4 Hz, 1H), 6.49-6.76 (m, 4H), 3.83 (s, 1H), 3.71 (s, 2H), 3.63 (s, 2H), 2.79 (d, J=5.6 Hz, 2H), 2.57-2.75 (m, 2H)

Example 15 Product Number TMI-105963

Name

N1-(3,5-dichloro-4-(4-hydroxy-3-((3-piperazin-1-methyl)phenoxy)phenyl)-N2-oxalyl hydroxylamine

(88) Structural Formula:

(89) ##STR00046##
Spectrum Data of Product:

(90) .sup.1H NMR (DMSO-d.sub.6) : 8.07-8.15 (m, 2H), 7.76 (s, 1H), 6.66-6.76 (m, 2H), 6.58 (dd, J=8.7, 3.1 Hz, 1H), 3.53 (s, 2H), 3.17 (s, 2H), 3.11 (br. s., 2H), 2.96 (s, 2H), 1.06 (t, J=7.0 Hz, 2H)

(91) Function and Effect of Examples:

(92) A. Mouse Experiment: TMI-105795 is Used as Test Compound in the Experiment

(93) I. Materials and Methods

(94) The experiment is conducted according to the procedures and details described below. To the best of our knowledge, no circumstances occurred during the course of the study that would have altered the quality and integrity of the data.

(95) (1) Substances to be Tested

(96) The tested compound (TMI-105795) is sub-packaged into dose volume and kept at 20 C. before use. All tested compounds (including solvents) are labeled as TDM-001-1, TDM-001-2, TDM-001-3 and TDM-001-4.

(97) (2) Animals in Experiments

(98) Female C3H mice (35-39 days after birth at arrival) were obtained from Beijing Vital River Laboratory Animal Technology Co., Ltd. The mice were selected for inclusion based upon acceptable clinical condition and body weight. Animals were housed randomly 5 mice per cage before grouping. All mice were kept at least one week in acclimation in the animal facility prior to any procedures. Animal identification number was labeled on the tail and cage tag respectively.

(99) (3) Animal Feeding

(100) Throughout the whole procedure of the experiment, the animals were housed in assigned groups (one mouse in each cage) in cages with soft bedding (corn cob or wood chips). Animals were conditioned to the same environment during the study course. Food and water were available ad libitum. Room humidity (50-70%) and temperature (21-25 C.) were maintained consistently. In addition, the study room was maintained on a 12-hour light/dark cycle.

(101) (4) Administration of Test Compound

(102) The test compound is TMI-105795 (solvent is polyethylene glycol/ethanol (30/70)), it is sub-packaged into dose volume and kept in 20 C. before use. It was topically administered together with solvent twice every day at 9:30 am and 4:00 pm.

(103) (5) Experimental Design

(104) TABLE-US-00001 Drug Test Number of Route of Dosing Level Treatment Group administration compound animals Treatment (l/mouse) Schedule 1 TDM-001-1 Solvent 5 topical 20 Twice/day, PG/EtOH (30/70) administration 5 days/week for 3 weeks re2 TDM-001-2 TMI-105795 5 topical 20 Twice/day, (0.005%(w/v)) administration 5 days/week for 3 weeks 3 TDM-001-3 TMI-105795 5 topical 20 Twice/day, (0.01%(w/v)) administration 5 days/week for 3 weeks 4 TDM-001-4 TMI-105795 5 topical 20 Twice/day, (0.05%(w/v)) administration 5 days/week for 3 weeks

(105) (6) Behavioral Observation

(106) Abnormal behaviors or hair growth were checked daily and data was carefully recorded during the experiment.

(107) (7) Body Weight

(108) During experiment, body weight of each animal was measured before administration.

(109) (8) Scoring

(110) Hair growth grade, peripheral hair growth, scaling, erythema will be scored on Monday, Wednesday and Friday from the beginning on the Day 1 to the termination of the experiment.

(111) II. Reagents, Consumables and Equipment

(112) (1) Test Compound (TMI-105795)

(113) (2) Eppendorf pipettes

(114) (3) Sterile 100 l tips

(115) (4) Electric shaver for small animal

(116) III. Experiment Procedures

(117) (1) Grouping

(118) All experimental mice were carefully dipped with the hair on the back of the rear leg with shaver. Care must be taken to avoid any irritation to the skin on the back of mouse. Skin color on the back was confirmed again. All mice must show pink skin, any mice that show signs of skin abrasion were excluded from the experiment. Mice were randomly divided into 5 groups and housed individually.

(119) (2) Administration Procedures

(120) Body weight of each mouse is recorded before administration every morning. 20 l corresponding solution is aspirated by pipette and applied to shaving area on the back of the mouse, and then the drug solution is uniformly applied to the back between rear legs with tips, about a region with a diameter of 1 cm. It is ensured that the administered region is the same one every day, and the mouse is put into the case after drying of liquid.

(121) (3) Scoring Procedure

(122) All mice are scored on Monday Wednesday and Friday each week from the beginning on the Day 1 to the termination of the study, using the following scoring guidelines:

(123) a. Hair Growth Grade (Administration Area):

(124) 0=No color change, or hair growth on the skin

(125) 1=Skin color changes uniformly over the administration area from pink to gray/black with no visible hair growth;

(126) 2=Appearance of short and sparse hair growth;

(127) 3=Full thick hair growth similar to the surrounding normal hair coat.

(128) b. Peripheral Hair Growth (Defined as Hair Growth that is in the Dipped Area, but not in the Administration Area):

(129) 0=No color change, or hair growth on the skin;

(130) 1=Skin color changes to gray/black with no visible hair growth;

(131) 2=Peripheral sparse hair growth;

(132) 3=Peripheral thick hair growth.

(133) c. Scaling:

(134) 0=none

(135) 1=mild

(136) 2=severe or peeling

(137) d. Erythema:

(138) 0=none

(139) 1=mild

(140) 2=severe

(141) e. Other Abnormalities (Precipitation of Test Compound at Dosing Site, Aberrant Hair Growth, Etc.)

(142) IV. Experimental Results

(143) (1) General Conditions of Mice During Experiment

(144) a. As shown in FIGS. 1(a)-(d), as compared with blank control group of solvent, the test compound (TMI-105795) shows a positive result in promoting hair growth after 3 weeks administration of test formulation at every dosing amount. There was no obvious hair growth in the solvent treatment group.
b. As shown in FIGS. 1(a)-(d), dose dependent effect is observed for different dose of the test compound obviously. Higher dose of test compound shows better effect in promoting hair growth compared with that of lower dose. As shown in FIG. 18, according to the scoring results of different mice in the experiment, no hair growth was observed for the blank control group of solvent, obvious difference was observed for the TDM-001-2 group (0.005% (w/v) compared with TDM-001-3 (i.e., 0.01% (w/v)) and TDM-001-4 (i.e., 0.05% (w/v)) groups.

(145) Obvious difference of hair growth rate was observed for the groups with different doses of test compound. Hair growth rate in TDM-001-2 group is far slower than the grow rate in TDM-001-3 and TDM-001-4.

(146) Specifically, for the mice in TDM-001-2 administration group, the skin became black on the day 12 of topical administration and hair growth was observed on the day 16. Continuous hair growth was observed on the following days.

(147) For the mice in TDM-001-3 administration group, the skin became black on day 2 of topical administration and hair growth was observed on the day 6. Continuous hair growth was observed on the following days.

(148) For the mice in TDM-001-4 topical administration group, the skin became black on day 2 of topical administration and hair growth was observed on the day 8. Continuous hair growth was observed on the following days.

(149) It can be seen from the experiment that, the onset of new hair growth and hair growth rate of mice in the low dose group are dearly delayed as compared to the high dose group. Although difference exists in hair growth progression with various doses, when the test compound dose reaches 0.01%, no obvious difference was observed in hair growth once the full coat hair grows to a certain level.

(150) c. As shown in FIG. 17, the body weight of all mice in each group increased slowly, indicating the test compound would not induce the loss of animal body weight.

(151) d. As shown in FIGS. 19 and 20, no significant findings were observed with scorings of scaling and erythema of the mice skin in all treatment groups.

(152) (2) Summary of Evaluation Results

(153) a. Positive result of promoting hair growth was observed for the test compound even at the lowest dosage in the experiment.

(154) b. Scaling and erythema were not observed in all treatment groups for all mice.

(155) Except the above observations, all mice showed normal skin color and no abnormal symptoms.

(156) V. Conclusion for Examples

(157) The test compound TMI-105795 significantly promotes the hair growth in C3H mice.

(158) C3H mouse's dorsal hair is known to have a time-synchronized hair growth cycle. From about 2.5 to 3.5 weeks old and 5 to 14 weeks old, the dorsal hairs are in the telogen phase (resting phase). From 0 to 2.0 weeks old and 4.0 to 4.5 weeks old, the dorsal hairs are in the anagen phase.

(159) In the experiment, the effect of the test compound (TMI-105795) on hair growth in C3H mice was tested. All doses of test compound (TMI-105795) showed positive results in promoting hair growth, when compared with solvent group, after 3 weeks topical administration of test formulation. At the same time, dose-dependent hair growth effect was clearly observed and related to the three different doses of the test compound.

(160) Therefore, this study showed the test compound (TMI-105795) could change hair growth cycle and promote hair growth.

(161) It is worth pointing out here, that the compounds in the other examples show similar effect. Redundant description is skipped here.

(162) B: Thyroid Receptor TR Agonist Assay

(163) I. Detailed experimental methods

(164) 1. Plating Cells

(165) (1) Two hours before cell seeding, pre-coat 96-well plates are added with 40 l poly-D-lysine, incubated at room temperature.

(166) (2) The residual poly-D-lysine is removed. On the 96-well plate, HEK-293 cells are plated at a density of 4.010.sup.4 cells/well in DMEM medium containing 10% FBS. The cells are cultured at 37 C. overnight.

(167) 2. Co-Transfection

(168) (1) The culture medium is removed 2 hours before transfection. 100 l of DMEM culture medium containing 10% FBS treated by activated carbon is added to each well, and incubated at 37 C. for 2 hours.

(169) (2) Preparation of plasmid DNA-XtremeGENE HP liposome complex.

(170) 1. Transfection reagent X-tremeGENE HP is warmed to room temperature and vortexed gently before use.

(171) 2. Opti-MEM I Reduced-Serum culture Medium is added in a sterile tube.

(172) 3. For each reaction system, 50 ng thyroid receptor (TR) plasmid DNA and 50 ng luciferase reporter gene plasmid DNA of thyroid hormone response element are added. Mixing it completely by gentle pipeting.

(173) 4. 0.2 l of transfection reagent X-tremeGENE HP is added to the diluted co-transfected DNA mixture. Mixing it completely by gentle pipeting.

(174) 5. The mixture is incubated at room temperature for 20 minutes to form plasmid DNA-XtremeGENE HP liposome complex.

(175) (3) 10 l of the complex is added to each well in a 96 well plate. Gently shaking the culturing plate to evenly distribute the complex.

(176) (4) The culture is incubated at 37 C. for 24 hours.

(177) 3. Agonist Compound Treatment:

(178) (1) Preparation of compound diluent: for each compound, half-log dilutions of each test compound is prepared using DMEM medium containing 10% charcoal stripped FBS (containing 0.1% DMSO), with a maximum concentration of 10 M of the test compound.

(179) (2) Removing the culture medium and adding the formulated compound diluent (the final DMSO concentration is 0.1%).

(180) (3) For blank control, DMEM medium containing 10% charcoal stripped FBS (containing 0.1% DMSO) is added.

(181) (4) Incubating at 37 C. for 24 hours.

(182) 4. Luciferase Reporter Gene Assay:

(183) (1) Removing the culture medium, adding 40 l phosphate (PBS) buffer to each well.

(184) (2) Adding 40 l Bright-Glo reagent to each well and vortexing for 30 seconds, followed by incubation in dark for 2 min.

(185) (3) Transferring 70 l cell lysate from each well into a white polystyrene 96-well plate.

(186) (4) Detecting the Luciferase signal intensity by Envision.

(187) II. Experimental Results (as shown in FIGS. 2-7)

(188) TABLE-US-00002 Compound Number EC50 (nM) 105798 179.4 TMI-105902 250.3 TMI-105903 136.4 TMI-105905 399.5 TMI-105965 95.97 TMI-105966 61.13 TMI-105906 925.4 TMI-105956 6521 TMI-105957 8757 TMI-105958 9701 TMI-105959 617.2 TMI-105960 6.478E7 TMI-105961 9243 TMI-105962 3719 TMI-105963 9577