Patent Application
20150265514
Unwanted hair is a common problem and subsequently there is an increasing demand for safe and efficient hair removal techniques. There are several traditional methods, including shaving, bleaching, plucking, waxing, use of chemical depilatories, and electrolysis, but these techniques have been limited by their pain, inconvenience, and poor long-term efficacy. Conditions such as hirsutism, a relatively frequent condition affecting about 4% of women, can severely interfere with personal and work activities. Temporary hair removal is a major component in the management of hirsute patients.
Photo-epilation is a common technique for long-term removal of unwanted hair, and typically involves thermal destruction of the hair follicle and its reproductive system (stems cells). However, despite considerable technical advances in this field, these devices still have the potential to cause injury when used improperly. Side effects after photoepilation are reported and include erythema and perifollicular oedema, which are common, as well as crusting, vesiculation, hypopigmentation and hyperpigmentation (depending on skin colour and other factors leading to excess heat production). Hair removal procedures can also damage hair follicles and lead to inflammation, which can in turn manifest severe skin conditions.
For example, since most hair removal procedures must be repeated periodically in order to remain free of unwanted hair, an alternative or complementary approach to hair removal is needed that, when combined with other methods of hair removal, for example, could enhance and prolong the removal effect, and reduce the need and frequency of hair removal. Agents that reduce or inhibit hair growth which can be applied to a treated area before, after and/or between procedures would be useful in prolonging epilation. Previous work with PPARγ modulators focused on using such modulators for hair growth, rather than inhibition of hair growth. Notably, such previous work on hair growth with such PPARγ modulators involved very low concentration of such modulators.
Thus, there remains a need for agents that retard hair regrowth after, e.g., the application of standard hair removal techniques
This disclosure is directed in part to methods for reducing mammalian hair growth comprising applying a composition comprising the compounds disclosed herein. For example, provided herein are methods for reducing mammalian hair growth, comprising selecting an area of mammalian skin for which a reduced rate of hair growth is desired; and applying a composition comprising N-acetyl-3-(4-aminophenyl)-2-methoxypropionic acid or a pharmaceutically acceptable salt or stereoisomer thereof, and a pharmaceutically acceptable excipient.
Also contemplated herein are methods of inhibiting hair growth after hair removal of an area of mammalian skin of a mammal, comprising topically applying a composition comprising the compounds disclosed herein (e.g., N-acetyl-3-(4-aminophenyl)-2-methoxypropionic acid) or a pharmaceutically acceptable salt or stereoisomer thereof, and a pharmaceutically acceptable excipient.
The disclosure is also directed in part to methods of reducing hair growth and hair shaft elongation in mammalian hair in need thereof, comprising topically applying a composition comprising the compounds disclosed herein. For example, provided herein are methods of reducing hair growth and hair shaft elongation in mammalian hair in need thereof comprising topically applying at least once daily for at least two days an effective amount of a composition comprising, e.g., N-acetyl-3-(4-aminophenyl)-2-methoxypropionic acid wherein after daily application for at least two days hair growth and hair shaft elongation are reduced in comparison with the appearance of hair without applying the composition.
The features and other details of the disclosure will now be more particularly described. Before further description of the present invention, certain terms employed in the specification, examples and appended claims are collected here. These definitions should be read in light of the remainder of the disclosure and understood as by a person of skill in the art. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by a person of ordinary skill in the art.
“Treating” includes any effect, e.g., lessening, reducing, modulating, or eliminating, that results in the improvement of the condition, disease, disorder and the like.
The term “alkenyl” as used herein refers to an unsaturated straight or branched hydrocarbon having at least one carbon-carbon double bond, such as a straight or branched group of 2-12, 2-10, or 2-6 carbon atoms, referred to herein as C2-C12alkenyl, C2-C10alkenyl, and C2-C6alkenyl, respectively. Exemplary alkenyl groups include, but are not limited to, vinyl, allyl, butenyl, pentenyl, hexenyl, butadienyl, pentadienyl, hexadienyl, 2-ethylhexenyl, 2-propyl-2-butenyl, 4-(2-methyl-3-butene)-pentenyl, etc.
The term “alkoxy” as used herein refers to an alkyl group attached to an oxygen (—O-alkyl-). Exemplary alkoxy groups include, but are not limited to, groups with an alkyl, alkenyl or alkynyl group of 1-12, 1-8, or 1-6 carbon atoms, referred to herein as C1-C12alkoxy, C1-C8alkoxy, and C1-C6alkoxy, respectively. Exemplary alkoxy groups include, but are not limited to methoxy, ethoxy, etc. Similarly, exemplary “alkenoxy” groups include, but are not limited to vinyloxy, allyloxy, butenoxy, etc.
The term “alkyl” as used herein refers to a saturated straight or branched hydrocarbon, such as a straight or branched group of 1-12, 1-10, or 1-6 carbon atoms, referred to herein as C1-C12alkyl, C1-C10alkyl, and C1-C6alkyl, respectively. Exemplary alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, 2-methyl-1-propyl, 2-methyl-2-propyl, 2-methyl-1-butyl, 3-methyl-1-butyl, 2-methyl-3-butyl, 2,2-dimethyl-1-propyl, 2-methyl-1-pentyl, 3-methyl-1-pentyl, 4-methyl-1-pentyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 2,2-dimethyl-1-butyl, 3,3-dimethyl-1-butyl, 2-ethyl-1-butyl, butyl, isobutyl, t-butyl, pentyl, isopentyl, neopentyl, hexyl, heptyl, octyl, etc. In certain embodiments, alkyl refers to C1-C6 alkyl. In certain embodiments, cycloalkyl refers to C3-C6cycloalkyl.
Alkyl, alkenyl and alkynyl groups can, in some embodiments, be optionally be substituted with or interrupted by at least one group selected from alkanoyl, alkoxy, alkyl, alkenyl, alkynyl, amido, amidino, amino, aryl, arylalkyl, azido, carbamate, carbonate, carboxy, cyano, cycloalkyl, ester, ether, formyl, halogen, haloalkyl, heteroaryl, heterocyclyl, hydroxyl, imino, ketone, nitro, phosphate, phosphonato, phosphinato, sulfate, sulfide, sulfonamido, sulfonyl and thiocarbonyl.
The term “alkynyl” as used herein refers to an unsaturated straight or branched hydrocarbon having at least one carbon-carbon triple bond, such as a straight or branched group of 2-12, 2-8, or 2-6 carbon atoms, referred to herein as C2-C12alkynyl, C2-C8alkynyl, and C2-C6alkynyl, respectively. Exemplary alkynyl groups include, but are not limited to, ethynyl, propynyl, butynyl, pentynyl, hexynyl, methylpropynyl, 4-methyl-1-butynyl, 4-propyl-2-pentynyl, and 4-butyl-2-hexynyl, etc.
The term “amide” or “amido” as used herein refers to a radical of the form —RaC(O)N(Rb)—, —RaC(O)N(Rb)Rc—, or —C(O)NRbRc, wherein Ra, Rb and Rc are each independently selected from alkoxy, alkyl, alkenyl, alkynyl, amide, amino, aryl, arylalkyl, carbamate, cycloalkyl, ester, ether, formyl, halogen, haloalkyl, heteroaryl, heterocyclyl, hydrogen, hydroxyl, ketone, and nitro. The amide can be attached to another group through the carbon, the nitrogen, Rb, Rc, or Ra. The amide also may be cyclic, for example Rb and Rc, Ra and Rb, or Ra and Rc may be joined to form a 3- to 12-membered ring, such as a 3- to 10-membered ring or a 5- to 6-membered ring. The term “carboxamido” refers to the structure —C(O)NRbRc.
The term “amine” or “amino” as used herein refers to a radical of the form —NRdRe, —N(Rd)Re—, or —ReN(Rd)Rf— where Rd, Re, and Rf are independently selected from alkoxy, alkyl, alkenyl, alkynyl, amide, amino, aryl, arylalkyl, carbamate, cycloalkyl, ester, ether, formyl, halogen, haloalkyl, heteroaryl, heterocyclyl, hydrogen, hydroxyl, ketone, and nitro. The amino can be attached to the parent molecular group through the nitrogen, Rd, Re or Rf. The amino also may be cyclic, for example any two of Rd, Re or Rf may be joined together or with the N to form a 3- to 12-membered ring, e.g., morpholino or piperidinyl. The term amino also includes the corresponding quaternary ammonium salt of any amino group, e.g., —[N(Rd)(Re)(Rf)]+. Exemplary amino groups include aminoalkyl groups, wherein at least one of Rd, Re, or Rf is an alkyl group.
The term “cycloalkoxy” as used herein refers to a cycloalkyl group attached to an oxygen.
The term “cycloalkyl” as used herein refers to a monovalent saturated or unsaturated cyclic, bicyclic, or bridged bicyclic hydrocarbon group of 3-12, 3-8, 4-8, or 4-6 carbons, referred to herein, e.g., as “C4-8cycloalkyl,” derived from a cycloalkane. Exemplary cycloalkyl groups include, but are not limited to, cyclohexanes, cyclohexenes, cyclopentanes, cyclopentenes, cyclobutanes and cyclopropanes. Cycloalkyl groups may be substituted with alkanoyl, alkoxy, alkyl, alkenyl, alkynyl, amido, amidino, amino, aryl, arylalkyl, azido, carbamate, carbonate, carboxy, cyano, cycloalkyl, ester, ether, formyl, halogen, haloalkyl, heteroaryl, heterocyclyl, hydroxyl, imino, ketone, nitro, phosphate, phosphonato, phosphinato, sulfate, sulfide, sulfonamido, sulfonyl and thiocarbonyl. Cycloalkyl groups can be fused to other cycloalkyl, aryl, or heterocyclyl groups. In certain embodiments, cycloalkyl refers to C3-C6 alkyl.
The terms “halo” or “halogen” or “Hal” as used herein refer to F, Cl, Br, or I.
The term “haloalkyl” as used herein refers to an alkyl group substituted with one or more halogen atoms.
The term “nitro” as used herein refers to the radical —NO2.
The term “phenyl” as used herein refers to a 6-membered carbocyclic aromatic ring. The phenyl group can also be fused to a cyclohexane or cyclopentane ring. Phenyl can be substituted with one or more substituents including alkanoyl, alkoxy, alkyl, alkenyl, alkynyl, amido, amidino, amino, aryl, arylalkyl, azido, carbamate, carbonate, carboxy, cyano, cycloalkyl, ester, ether, formyl, halogen, haloalkyl, heteroaryl, heterocyclyl, hydroxyl, imino, ketone, nitro, phosphate, phosphonato, phosphinato, sulfate, sulfide, sulfonamido, sulfonyl and thiocarbonyl.
The term “pharmaceutically acceptable carrier” or “pharmaceutically acceptable excipient” as used herein refers to any and all solvents, dispersion media, coatings, isotonic and absorption delaying agents, and the like, that are compatible with pharmaceutical administration. The use of such media and agents for pharmaceutically active substances is well known in the art. The compositions may also contain other active compounds providing supplemental, additional, or enhanced therapeutic functions.
The term “pharmaceutical composition” as used herein refers to a composition comprising at least one compound as disclosed herein formulated together with one or more pharmaceutically acceptable carriers.
“Individual,” “patient,” or “subject” are used interchangeably and include to any animal, including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, swine, cattle, sheep, horses, or primates, and most preferably humans. The compounds disclosed herein can be administered to a mammal, such as a human, but can also be other mammals such as an animal in need of veterinary treatment, e.g., domestic animals (e.g., dogs, cats, and the like), farm animals (e.g., cows, sheep, pigs, horses, and the like) and laboratory animals (e.g., rats, mice, guinea pigs, and the like). “Modulation” includes antagonism (e.g., inhibition), agonism, partial antagonism and/or partial agonism.
In the present specification, the term “therapeutically effective amount” means the amount of the subject compound that will elicit the biological or medical response of a tissue, system, animal or human that is being sought by the researcher, veterinarian, medical doctor or other clinician. The compounds disclosed herein are administered in therapeutically effective amounts to treat a disease. Alternatively, a therapeutically effective amount of a compound is the quantity required to achieve a desired therapeutic and/or prophylactic effect, such as an amount which results in, for example, hair growth retardation.
The term “pharmaceutically acceptable salt(s)” as used herein refers to salts of acidic or basic groups that may be present in compounds used in the present compositions. Compounds included in the present compositions that are basic in nature are capable of forming a wide variety of salts with various inorganic and organic acids. The acids that may be used to prepare pharmaceutically acceptable acid addition salts of such basic compounds are those that form non-toxic acid addition salts, i.e., salts containing pharmacologically acceptable anions, including but not limited to malate, oxalate, chloride, bromide, iodide, nitrate, sulfate, bisulfate, phosphate, acid phosphate, isonicotinate, acetate, lactate, salicylate, citrate, tartrate, oleate, tannate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisinate, fumarate, gluconate, glucuronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate and pamoate (i.e., 1,1′-methylene-bis-(2-hydroxy-3-naphthoate)) salts. Compounds included in the present compositions that include an amino moiety may form pharmaceutically acceptable salts with various amino acids, in addition to the acids mentioned above. Compounds included in the present compositions that are acidic in nature are capable of forming base salts with various pharmacologically acceptable cations. Examples of such salts include alkali metal or alkaline earth metal salts and, particularly, calcium, magnesium, sodium, lithium, zinc, potassium, and iron salts.
The compounds of the disclosure may contain one or more chiral centers and/or double bonds and, therefore, exist as stereoisomers, such as geometric isomers, enantiomers or diastereomers. The term “stereoisomers” when used herein consist of all geometric isomers, enantiomers or diastereomers. These compounds may be designated by the symbols “R” or “S,” depending on the configuration of substituents around the stereogenic carbon atom. The present invention encompasses various stereoisomers of these compounds and mixtures thereof. Stereoisomers include enantiomers and diastereomers. Mixtures of enantiomers or diastereomers may be designated “(±)” in nomenclature, but the skilled artisan will recognize that a structure may denote a chiral center implicitly.
Individual stereoisomers of compounds of the present invention can be prepared synthetically from commercially available starting materials that contain asymmetric or stereogenic centers, or by preparation of racemic mixtures followed by resolution methods well known to those of ordinary skill in the art. These methods of resolution are exemplified by (1) attachment of a mixture of enantiomers to a chiral auxiliary, separation of the resulting mixture of diastereomers by recrystallization or chromatography and liberation of the optically pure product from the auxiliary, (2) salt formation employing an optically active resolving agent, or (3) direct separation of the mixture of optical enantiomers on chiral chromatographic columns. Stereoisomeric mixtures can also be resolved into their component stereoisomers by well known methods, such as chiral-phase gas chromatography, chiral-phase high performance liquid chromatography, crystallizing the compound as a chiral salt complex, or crystallizing the compound in a chiral solvent. Stereoisomers can also be obtained from stereomerically-pure intermediates, reagents, and catalysts by well known asymmetric synthetic methods.
Geometric isomers can also exist in the compounds of the present invention. The symbol ═ denotes a bond that may be a single, double or triple bond as described herein. The present invention encompasses the various geometric isomers and mixtures thereof resulting from the arrangement of substituents around a carbon-carbon double bond or arrangement of substituents around a carbocyclic ring. Substituents around a carbon-carbon double bond are designated as being in the “Z” or “E” configuration wherein the terms “Z” and “E” are used in accordance with IUPAC standards. Unless otherwise specified, structures depicting double bonds encompass both the “E” and “Z” isomers.
Substituents around a carbon-carbon double bond alternatively can be referred to as “cis” or “trans,” where “cis” represents substituents on the same side of the double bond and “trans” represents substituents on opposite sides of the double bond. The arrangement of substituents around a carbocyclic ring are designated as “cis” or “trans.” The term “cis” represents substituents on the same side of the plane of the ring and the term “trans” represents substituents on opposite sides of the plane of the ring. Mixtures of compounds wherein the substituents are disposed on both the same and opposite sides of plane of the ring are designated “cis/trans.”
The compounds disclosed herein can exist in solvated as well as unsolvated forms with pharmaceutically acceptable solvents such as water, ethanol, and the like, and it is intended that the invention embrace both solvated and unsolvated forms. In one embodiment, the compound is amorphous. In one embodiment, the compound is a polymorph. In another embodiment, the compound is in a crystalline form.
The invention also embraces isotopically labeled compounds which are identical to those recited herein, except that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can be incorporated into compounds disclosed herein include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine and chlorine, such as 2H, 3H, 13C, 14C, 15N, 18O, 17O, 31P, 32P, 35S, 18F, and 36Cl, respectively.
Certain isotopically-labeled disclosed compounds (e.g., those labeled with 3H and 14C) are useful in compound and/or substrate tissue distribution assays. Tritiated (i.e., 3H) and carbon-14 (i.e., 14C) isotopes are particularly preferred for their ease of preparation and detectability. Further, substitution with heavier isotopes such as deuterium (i.e., 2H) may afford certain therapeutic advantages resulting from greater metabolic stability (e.g., increased in vivo half-life or reduced dosage requirements) and hence may be preferred in some circumstances. Isotopically labeled compounds of the invention can generally be prepared by following procedures analogous to those disclosed in the e.g., Examples herein by substituting an isotopically labeled reagent for a non-isotopically labeled reagent.
The term “prodrug” refers to compounds that are transformed in vivo to yield a disclosed compound or a pharmaceutically acceptable salt, hydrate or solvate of the compound. The transformation may occur by various mechanisms, such as through hydrolysis in blood. For example, if a compound disclosed herein or a pharmaceutically acceptable salt, hydrate or solvate of the compound contains a carboxylic acid functional group, a prodrug can comprise an ester formed by the replacement of the hydrogen atom of the acid group with a group such as (C1-C8)alkyl, (C2-C12)alkanoyloxymethyl, 1-(alkanoyloxy)ethyl having from 4 to 9 carbon atoms, 1-methyl-1-(alkanoyloxy)-ethyl having from 5 to 10 carbon atoms, alkoxycarbonyloxymethyl having from 3 to 6 carbon atoms, 1-(alkoxycarbonyloxy)ethyl having from 4 to 7 carbon atoms, 1-methyl-1-(alkoxycarbonyloxy)ethyl having from 5 to 8 carbon atoms, N-(alkoxycarbonyl)aminomethyl having from 3 to 9 carbon atoms, 1-(N-(alkoxycarbonyl)amino)ethyl having from 4 to 10 carbon atoms, 3-phthalidyl, 4-crotonolactonyl, gamma-butyrolacton-4-yl, di-N,N—(C1-C2)alkylamino(C2-C3)alkyl (such as β-dimethylaminoethyl), carbamoyl-(C1-C2)alkyl, N,N-di(C1-C2)alkylcarbamoyl-(C1-C2)alkyl and piperidino-, pyrrolidino- or morpholino(C2-C3)alkyl.
Similarly, if a compound disclosed herein contains an alcohol functional group, a prodrug can be formed by the replacement of the hydrogen atom of the alcohol group with a group such as (C1-C6)alkanoyloxymethyl, 1-((C1-C6)alkanoyloxy)ethyl, 1-methyl-1-((C1-C6)alkanoyloxy)ethyl (C1-C6)alkoxycarbonyloxymethyl, N—(C1-C6)alkoxycarbonylaminomethyl, succinoyl, (C1-C6)alkanoyl, α-amino(C1-C4)alkanoyl, arylacyl and α-aminoacyl, or α-aminoacyl-α-aminoacyl, where each α-aminoacyl group is independently selected from the naturally occurring L-amino acids, P(O)(OH)2, —P(O)(O(C1-C6)alkyl)2 or glycosyl (the radical resulting from the removal of a hydroxyl group of the hemiacetal form of a carbohydrate).
If a compound disclosed herein incorporates an amine functional group, a prodrug can be formed by the replacement of a hydrogen atom in the amine group with a group such as R-carbonyl, RO-carbonyl, NRR′-carbonyl where R and R′ are each independently (C1-C10)alkyl, (C3-C7)cycloalkyl, benzyl, or R-carbonyl is a natural α-aminoacyl or natural α-aminoacyl-natural α-aminoacyl, —C(OH)C(O)OY1 wherein Y1 is H, (C1-C6)alkyl or benzyl, —C(OY2)Y3 wherein Y2 is (C1-C4) alkyl and Y3 is (C1-C6)alkyl, carboxy(C1-C6)alkyl, amino(C1-C4)alkyl or mono-N— or di-N,N—(C1-C6)alkylaminoalkyl, —C(Y4)Y5 wherein Y4 is H or methyl and Y5 is mono-N— or di-N,N—(C1-C6)alkylamino, morpholino, piperidin-1-yl or pyrrolidin-1-yl.
Compounds contemplated for use in one or more of the disclosed methods are represented by formula I, as depicted below. Also contemplated herein are compositions that include a compound represented by formula I and e.g., a pharmaceutically or cosmetically acceptable carrier or excipient.
wherein X is C1-C3alkylene, optionally substituted with one, two or three substituents selected from halogen or hydroxyl;
R1 is selected from the group consisting of C1-C6alkyl, C3-C6cycloalkyl, C2-C6alkenyl, and C2-C6alkynyl;
R2 is selected from the group consisting of hydrogen and C1-C6alkyl;
R3 is independently selected, for each occurrence from the group consisting of hydrogen, C1-C6alkoxy, C1-C6alkyl, cyano, C3-C6cycloalkyl, halogen, hydroxyl, and nitro;
R4 is selected from the group consisting of hydrogen and C1-C6alkyl;
R5 is C1-C6alkyl;
or pharmaceutically acceptable salts or N-oxides thereof.
In one embodiment, R1 can be C1-C6alkyl, such as methyl. In one embodiment, R2 can be hydrogen. In another embodiment, R3 can be selected from the group consisting of hydrogen, C1-C6alkyl, halogen, and hydroxyl. In a further embodiment, R3 can be hydrogen. In one embodiment, R4 and R5 can each be C1-C6alkyl. In another embodiment, R4 may be hydrogen and R5 may be methyl. In one embodiment, X may be (CH2)n wherein n is 1 or 2, such as 1.
In another embodiment, —NR2—COR1 can be in the meta position relative to X as shown in formula II.
In another embodiment, —NR2—COR1 can be in the para position relative to X as shown in formula III.
The disclosure provides, at least in part, compounds represented by formula IV, as depicted below. Also contemplated herein are compositions that include a compound represented by formula IV and e.g., a pharmaceutically acceptable carrier.
wherein R1 is selected from the group consisting of C1-C6alkyl, C3-C6cycloalkyl, C2-C6alkenyl, and C2-C6alkynyl;
R2 is selected from the group consisting of hydrogen and C1-C6alkyl;
R3 is independently selected, for each occurrence from the group consisting of hydrogen, C1-C6alkoxy, C1-C6alkyl, cyano, C3-C6cycloalkyl, halogen, hydroxyl, and nitro;
R5 is hydrogen or C1-C6alkyl;
or pharmaceutically acceptable salts or N-oxides thereof.
Compounds of Formula V are also contemplated as shown below, as well as compositions that include a compound represented by formula V and e.g., a pharmaceutically acceptable carrier.
wherein R1 is selected from the group consisting of C1-C6alkyl, C3-C6cycloalkyl, C2-C6alkenyl, and C2-C6alkynyl;
R3 is independently selected, for each occurrence from the group consisting of hydrogen, C1-C6alkoxy, C1-C6alkyl, cyano, C3-C6cycloalkyl, halogen, hydroxyl, and nitro;
R4 is selected from the group consisting of hydrogen and C1-C6alkyl;
R5 is hydrogen or C1-C6alkyl; and
A is a fused five or six membered heterocycle;
or pharmaceutically acceptable salts or N-oxides thereof.
In one embodiment, R1 can be C1-C6alkyl, such as methyl. In another embodiment, R1 and R3 can each be C1-C6alkyl, such as methyl. In one embodiment, R2 can be hydrogen.
In some embodiments, a compound can be represented by
wherein p is 1 or 2;
R1 is selected from the group consisting of C1-C6alkyl, C3-C6cycloalkyl, C2-C6alkenyl, and C2-C6alkynyl;
R4 and R8 are each independently selected from the group consisting of hydrogen and C1-C6alkyl;
or pharmaceutically acceptable salts or N-oxides thereof.
Contemplated compounds, and pharmaceutical compositions, comprising at least one compound, may be selected from the group consisting of: N-acetyl-(R)-(−)-3-(4-aminophenyl)-2-methoxypropionic acid (Compound A), N-acetyl-(S)-(−)-3-(4-aminophenyl)-2-methoxypropionic acid (Compound B), racemic N-acetyl-(S)-(−)-3-(4-aminophenyl)-2-methoxypropionic acid (compound AB);
4-acetamino-N-hydroxy-2-methoxybenzamide; 1-acetyl-6-methoxy-1,2,3,4-tetrahydroquinoline-5-carboxylic acid, 5-acetamido-2-hydroxybenzoic acid (e.g., acetylated 5-aminosalicyclic acid) or pharmaceutically acceptable salts or N-oxides thereof. The present disclosure also provides pharmaceutical compositions comprising compounds as disclosed herein formulated together with one or more pharmaceutically or cosmetically acceptable carriers. These formulations include those suitable for oral, rectal, topical, buccal and parenteral (e.g., subcutaneous, intramuscular, intradermal, or intravenous) administration, or for topical use, e.g. as part of a composition suitable for applying topically to skin. Although the most suitable form of administration in any given case will depend on the degree and severity of the condition being treated and on the nature of the particular compound being used.
Additional compounds contemplated for use in one or more of the disclosed methods include compounds represented by formula VIII, or a pharmaceutically acceptable salt, enantiomer or stereoisomer thereof:
wherein:
R1 and R2, are each independently selected from the group consisting of H and C1-6 alkyl; or R1 and R2 together with the nitrogen atom they are bonded to form an aromatic or aliphatic ring with 5 or 6 atoms which may be optionally substituted;
Y and Z are each independently selected from the group consisting of H, OH, COOH, —OR3, —CH(OR3)COOH; and
R3 is selected from the group consisting of H, phenyl, benzyl, vinyl, allyl, C1-6 alkyl or C1-6 alkyl substituted by one or more halogens.
In an embodiment, Y may be H or COOH. For example, Y may be H and Z may be CH(OR3)COOH, or Y may be COOH and Z maybe —OR3. In some embodiments, R3 may be methyl, ethyl, n-propyl, or isopropyl.
In other embodiments, the NR1R2 moiety may be in the 4′ position or may be in the 3′ position. In certain embodiments, R1 and R2 are H.
Exemplary compounds also include those represented by formulas IXa or IXb or a pharmaceutically acceptable salt, enantiomer or stereoisomer of:
wherein:
R1 and R2 are each independently selected from the group consisting of H and C1-6 alkyl; or R1 and R2 together, with the nitrogen atom they are bonded to, form an aromatic or aliphatic ring with 5 or 6 atoms;
R6 is selected from the group consisting of: —NHOH, OH, and —OR9;
R9 is C1-6 alkyl;
R4 is selected from H, phenyl, benzyl, vinyl, allyl, C1-6 alkyl or C1-6 alkyl substituted by one or more halogens;
R5 and R7 are each independently hydrogen or halo; or
R4 and R5, or R4 and R6 together, form a fused heterocyclic ring with 5 or 6 atoms, optionally substituted with halo or C1-6 alkyl; and
A is a fused heterocyclic ring; or a pharmaceutically acceptable salt thereof.
In certain embodiments, the NR1R2 moiety of formula IIa may be in the 4′ position or may be in the 3′ position. In certain embodiments, R1 and R2 are H.
R9, in some embodiments, may be methyl, ethyl, n-propyl, or isopropyl.
In some embodiments a compound can be represented by
wherein p is 1 or 2, R6 is OH or —OR9, wherein R9 is defined above, and R10, independently for each occurrence, is selected from the group consisting of H, halo, or C1-6 alkyl, e.g. methyl or ethyl.
Exemplary compounds contemplated herein include:
or a pharmaceutically acceptable salt thereof.
In some embodiments, contemplated compounds include: 4-amino-N-hydroxy-2-methoxybenzamide (compound 13); 6-methoxy quinoline-5-carboxylic acid (compound 36); 6-methoxy-1,2,3,4-tetrahydroquinoline-5-carboxylic acid (compound 37); 5-diisopropylaminosalicylic acid (compound 38).
Other exemplary compounds include those represented by:
Compounds contemplated herein include racemic mixtures, and enantiomers of compounds, for example: (±)-2-hydroxy-3-(3′-aminophenyl) propionic acid (compound 20); (±)-2-methoxy-2-(4′-aminophenyl) acetic acid (compound 23); (±)-2-ethoxy-2-(3′-aminophenyl) acetic acid (compound 32); (±)-2-ethoxy-2-(4′-aminophenyl) acetic acid (compound 33); (±)-2-methoxy-3-(4′-aminophenyl) propionic acid (compound 34) “±34” (racemic form); (±)-2-ethoxy-3-(4′-aminophenyl) propionic acid (compound 39); (±)-2-ethoxy-3-(3′-aminophenyl) propionic acid (compound 40).
For example, the compounds used in the methods disclosed herein can be enantiomers of the following racemic mixtures: (R,S)-2-hydroxy-2-(3-aminophenyl)acetic acid (compound 10); (R,S)-2-hydroxy-2-(4-aminophenyl)acetic acid (compound 11); (R,S)-2-hydroxy-3-(4′-aminophenyl)propionic acid (compound 21); (R,S)-2-methoxy-2-(3′-aminophenyl)acetic acid (compound 22); (R,S)-2-methoxy-3-(3′-aminophenyl)propionic acid (compound 35); (R,S)-2-methoxy-3-(4-aminophenyl)propionic acid (compound 34), as well as enantiomers, e.g.: (+) 2-S-methoxy-3-(4-aminophenyl)propionic acid (compound 34); (−) 2-R-methoxy-3-(4-aminophenyl)propionic acid (compound 34).
Other racemic type mixtures of compounds contemplated include: e.g. (±)-2-hydroxy-2-(3′-aminophenyl)acetic acid (compound 10); (±)-2-hydroxy-2-(4′-aminophenyl)acetic acid (compound 11); (±)-2-hydroxy-3-(4′-aminophenyl)propionic acid (compound 21) and (±)-2-methoxy-2-(3′-aminophenyl)acetic acid (compound 22).
Further compounds contemplated for use in the disclosed methods: 5-aminosalicylo-hydroxamic acid (compound 5); 3-dimethylaminosalicylic acid (compound 6); 2-methoxy-4-aminobenzoic acid (compound 7); 2-methoxy-5-aminobenzoic acid (compound 8); 5-methylaminosalicylic acid (compound 9); 4-methylaminosalicylic acid (compound 12); 4-acetylaminosalicylic acid (compound 16); 2-ethoxy-4-aminobenzoic acid (compound 18); 2-ethoxy-5-aminobenzoic acid (compound 19); 4-dimethylaminosalicylic acid (compound 24); 2-ethoxy-4-aminobenzoylhydroxamic acid (compound 25); 6-hydroxyquinoline-5-carboxylic acid (compound 27); 2-(2-propyl)oxy-4-aminobenzoic acid (compound 30); 4-(1-piperazinyl)salicylic acid (compound 41); (R,S) 5-oxa-quinoline-6-carboxylic acid (compound 15); 6-methoxy quinoline-5-carboxylic acid (compound 36); 6-methoxy-1,2,3,4-tetrahydroquinoline-5-carboxylic acid (compound 37); 5-diisopropylaminosalicylic acid (compound 38); and 4-diisopropylaminosalicylic acid (compound 42).
Methods for making contemplated compounds may be found for example in WO2007/010516 and WO2007/010514, each hereby incorporated by reference in their entirety.
The disclosure is directed, at least in part, to methods for reducing mammalian hair growth in an area of mammalian skin using a composition comprising a compound disclosed herein. For example, provided herein are methods for reducing mammalian hair growth by selecting an area of mammalian skin for which a reduced rate of hair growth is desired; and applying a composition comprising N-acetyl-3-(4-aminophenyl)-2-methoxypropionic acid or a pharmaceutically acceptable salt or stereoisomer thereof, and a pharmaceutically acceptable excipient.
In certain embodiments, for example, after applying, a disclosed composition significantly reduces hair shaft elongation. In other embodiments, for example, applying a disclosed composition occurs before and/or after substantial hair removal (for example, via laser hair removal, a depilatory cream hair remover, a wax hair removal, electrolysis hair removal, and/or a razor hair removal from the area). In another embodiment, for example, applying a disclosed composition occurs after the substantial hair removal.
The disclosure further provides for methods of inhibiting hair growth after hair removal of an area of mammalian skin of a mammal (e.g. human skin), comprising topically applying a composition comprising a compound disclosed herein. For example, the disclosure provides for methods of inhibiting hair growth after hair removal of an area of mammalian skin of a mammal, comprising topically applying a composition comprising N-acetyl-3-(4-aminophenyl)-2-methoxypropionic acid or a pharmaceutically acceptable salt or stereoisomer thereof, and a pharmaceutically acceptable excipient. For example, provided herein is a method that prolongs depilation, and/or protects hair follicle cells from cell death.
In certain embodiments, a disclosed composition comprises N-acetyl-(R)-(−)-3-(4-aminophenyl)-2-methoxypropionic acid. In other embodiments, a disclosed composition comprises a concentration of at least about 1 mM of the N-acetyl-3-(4-aminophenyl)-2-methoxypropionic acid. For example, a disclosed composition may comprise a concentration of at least about 1 mM, about 2 mM, about 3 mM, about 4 mM, about 5 mM, about 6 mM, about 7 mM, about 8 mM, about 9 mM or about 10 mM. In another embodiment, a disclosed composition comprises a concentration of at least about 1 mM to about 10 mM of a disclosed compound, for example, N-acetyl-3-(4-aminophenyl)-2-methoxypropionic acid. For example, a disclosed composition may comprise a concentration of at least about 1.5 mM to about 9.5 mM, about 2 mM to about 9 mM, about 2.5 mM to about 8.5 mM, about 3 mM to about 8 mM, about 3.5 mM to about 7.5 mM, about 4 mM to about 7 mM or about 4.5 mM to about 6.5 mM of a disclosed compound.
For example, disclosed methods may include topically applying a composition having a high concentration of a disclosed compound, e.g., that provides an effective amount significantly greater than the amount necessary to, e.g., induce hair growth. For example, such high concentration of such a composition may include about 1 mM to about 1000 mM or more of a disclosed compound, e.g. about 1 mM to about 100 mM, about 10 mM to about 100 mM, or about 10 mM to about 50 mM.
The compounds disclosed herein may be administered to subjects (animals and/or humans) in need of such treatment in dosages that will provide optimal pharmaceutical efficacy. It will be appreciated that the dose required for use in any particular application will vary from patient to patient, not only with the particular compound or composition selected, but also with the route of administration, the nature of the condition being treated, the age and condition of the patient, concurrent medication or special diets then being followed by the patient, and other factors which those skilled in the art will recognize, with the appropriate dosage ultimately being at the discretion of the attendant physician. For treating clinical conditions and diseases noted above, a disclosed compound or composition may be administered orally, topically, parenterally, by inhalation spray or rectally in dosage unit formulations containing conventional non-toxic pharmaceutically acceptable carriers, adjuvants and vehicles. The term parenteral as used herein includes subcutaneous injections, intravenous, intramuscular, intrasternal injection or infusion techniques.
Generally, the amount of active component considered to be therapeutically effective will depend on variables such as the type and extent of disease or indication to be treated, the overall health status of the particular patient, the relative biological efficacy of the compounds, formulation of compounds, the presence and types of excipients in the formulation, and the route of administration. The initial dosage administered may be increased beyond the upper level in order to rapidly achieve the desired blood-level or tissue level, or the initial dosage may be smaller than the optimum and the daily dosage may be progressively increased during the course of treatment depending on the particular situation. Dosing frequency can vary, depending on factors such as route of administration, dosage amount and the disease condition being treated. Exemplary dosing frequencies are at least once per day, at least once per week and at least once every two weeks.
Contemplated formulations or compositions comprise a disclosed compound and typically may also include a pharmaceutically acceptable carrier or expicient.
Contemplated compositions may be administered by various means, depending on their intended use, as is well known in the art. Formulations disclosed herein may be administered topically. These formulations may be prepared by conventional means, and, if desired, disclosed compositions may be mixed with any conventional additive, such as an excipient, a binder, a disintegrating agent, a lubricant, a corrigent, a solubilizing agent, a suspension aid, an emulsifying agent or a coating agent.
In formulations disclosed herein, wetting agents, emulsifiers and lubricants, such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, release agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants may be present in the formulated agents. Methods of preparing these formulations include the step of bringing into association compositions disclosed herein with the carrier and, optionally, one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately bringing into association agents with liquid carriers, or finely divided solid carriers, or both, and then, if necessary, shaping the product.
Suspensions, in addition to the subject composition, may contain suspending agents as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, and mixtures thereof.
Dosage forms for transdermal or topical administration of a subject composition include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches and inhalants. The active component may be mixed under sterile conditions with a pharmaceutically acceptable carrier, and with any preservatives, buffers, or propellants which may be required.
The ointments, pastes, creams and gels may contain, in addition to a subject composition, excipients, such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide, or mixtures thereof.
Powders and sprays may contain, in addition to a subject composition, excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powder, or mixtures of these substances. Sprays may additionally contain customary propellants, such as chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons, such as butane and propane.
Compositions and compounds of the present disclosure may alternatively be administered by aerosol. This is accomplished by preparing an aqueous aerosol, liposomal preparation or solid particles containing the compound. A non-aqueous (e.g., fluorocarbon propellant) suspension could be used. Sonic nebulizers may be used because they minimize exposing the agent to shear, which may result in degradation of the compounds contained in the subject compositions.
Ordinarily, an aqueous aerosol is made by formulating an aqueous solution or suspension of a subject composition together with conventional pharmaceutically acceptable carriers and stabilizers. The carriers and stabilizers vary with the requirements of the particular subject composition, but typically include non-ionic surfactants (Tweens, Pluronics, or polyethylene glycol), innocuous proteins like serum albumin, sorbitan esters, oleic acid, lecithin, amino acids such as glycine, buffers, salts, sugars or sugar alcohols. Aerosols generally are prepared from isotonic solutions.
Pharmaceutical compositions disclosed herein may be suitable for parenteral administration comprise a subject composition in combination with one or more pharmaceutically-acceptable sterile isotonic aqueous or non-aqueous solutions, dispersions, suspensions or emulsions, or sterile powders which may be reconstituted into sterile injectable solutions or dispersions just prior to use, which may contain antioxidants, buffers, bacteriostats, solutes which render the formulation isotonic with the blood of the intended recipient or suspending or thickening agents.
Examples of suitable aqueous and non-aqueous carriers which may be employed in the pharmaceutical compositions disclosed herein include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, and the like), and suitable mixtures thereof, vegetable oils, such as olive oil, and injectable organic esters, such as ethyl oleate and cyclodextrins. Proper fluidity may be maintained, for example, by the use of coating materials, such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants. The efficacy of treatment with the subject compositions may be determined in a number of fashions known to those of skill in the art.
Throughout the description, where compositions are described as having, including, or comprising specific components, it is contemplated that compositions also consist essentially of, or consist of, the recited components. Similarly, where processes are described as having, including, or comprising specific process steps, the processes also consist essentially of, or consist of, the recited processing steps. Except where indicated otherwise, the order of steps or order for performing certain actions are immaterial so long as the invention remains operable. Moreover, unless otherwise noted, two or more steps or actions may be conducted simultaneously.
The compounds disclosed herein can be prepared in a number of ways well known to one skilled in the art of organic synthesis. More specifically, compounds disclosed herein may be prepared using the reactions and techniques described herein. In the description of the synthetic methods described below, it is to be understood that all proposed reaction conditions, including choice of solvent, reaction atmosphere, reaction temperature, duration of the experiment and workup procedures, can be chosen to be the conditions standard for that reaction, unless otherwise indicated. It is understood by one skilled in the art of organic synthesis that the functionality present on various portions of the molecule should be compatible with the reagents and reactions proposed. Substituents not compatible with the reaction conditions will be apparent to one skilled in the art, and alternate methods are therefore indicated. The starting materials for the examples are either commercially available or are readily prepared by standard methods from known materials.
To (R)-(−)-3-(4-aminophenyl)-2-methoxypropionic acid (40 g) in a 0.5 L glass reactor was added ethyl acetate (80 g) and acetic anhydride (62.8 g). The mixture was stirred at 90° C. for 1 hour. Upon cooling, the solvent was removed by vacuum distillation, providing an oily residue. To this residue was added water (120 g) and ethyl acetate (120 g). After stirring for 10 min at 35° C., the layers were separated and the aqueous layer discarded. The organic layer solvent was removed by vacuum distillation. Acetone (120 g) was then added and the resulting mixture was warmed until dissolution was complete. The solution was cooled to 0° C., and the product precipitated which was collected by filtration. The solid was rinsed with acetone (20 g) and dried at 65° C. to afford 26 g of the title compound.
The aim of the present study is to determine the effects of Compound A on stem cell markers in the hair follicle by evaluating the expression of stem call markers K15 and K19.
Normal human scalp skin was obtained from 6 women undergoing routine face-lift surgery after informed consent. All experiments were performed according to Helsinki guidelines, with appropriate ethics committee approval. Details on specimen origin are listed in Table 1.
Normally pigmented anagen VI hair follicles (HFs) (grey/white HFs were excluded from the study) were microdissected from normal human scalp skin and organ cultured following the Philpott model. Compound A or vehicle was administered once for each change of medium (i.e. every 48 h/72 h). An overview of the experimental procedures is presented in
To investigate keratin K15 expression, the tyramide signal amplification method was used as previously described (Kloepper et al., 2008). Briefly, acetone fixed cryosections were washed three times for 5 min using TNT (Tris-HCL NaCl Tween) buffer (0.1 mol/1 Tris-HCl, pH 7.5; containing 0.15 mol/1 NaCl and 0.05% Tween 20). Next, horseradish peroxidase was blocked by washing with 3% H2O2 in phosphate-buffered saline (PBS) for 15 min. Preincubation was performed with the incubation of avidin and biotin for 15 min and 5% goat normal serum in TNT for 30 min with washing steps in between. Mouse anti-human K15 (clone LHK15, Chemicon, Billerica, USA) was diluted in TNT and incubated overnight at 4° C. followed by a biotinylated secondary antibody goat anti-mouse (1:200 in TNT) for 45 min at RT. Next, streptavidin horseradish peroxidase (TSA kit; Perkin-Elmer, Boston, Mass., USA) was administered (1:100 in TNT) for 30 min at RT. The reaction was amplified by FITC-tyramide amplification reagent at RT for 5 min (1:50 in amplification diluent provided with the kit). The intensity of the immunostaining was quantified by ImageJ software (National Institutes of Health). The staining intensity of defined reference regions in the HF was measured and compared between control and N Compound A-treated groups. The percentage of K15-positive cells in comparison to the total amount of cells in the ORS was calculated.
A previously described protocol was used to investigate K19 expression (Kloepper et al., 2008). Briefly, acetone-fixed cryosections were pre-treated with goat serum (10% in Tris-buffered saline, Dako). The sections were incubated first with primary antibodies against K19 (mouse anti-human: K19-1:10; overnight, at 4° C.; PROGEN, Heidelberg, Germany;) and then with FITC-labeled goat anti-mouse (1:200 in TBS, for 45 min, RT, Jackson ImmunoResearch) immunoglobulins as secondary antibodies. Counterstaining was performed with DAPI (Boehringer Mannheim, Mannheim, Germany). The intensity of this immunostaining was quantified by ImageJ software (National Institutes of Health). The staining intensity of defined reference regions in the HF was measured and compared between control and Compound A-treated groups. The percentage of K19-positive cells in comparison to the total amount of cells in the ORS was calculated.
Statistical analysis was performed using a two-tailed Student's t-test for unpaired samples. For meta-analysis purposes, a total of six assays (each with HFs from a different female individual) were run. For keratin 19, only 5 assays were available for analysis since the number of usable hair follicle sections for quantitative immunohistomorphometry did not suffice to also run this parameter. In order to avoid data distortion by individual experiments, rigid exclusion criteria were defined that allowed to exclude one individual experiment (out of 5-6) per read-out parameter. Since these exclusion criteria differed for each study parameter, different experiments (i.e. one out of 6, and one out of 5 in the case of keratin 19 analysis) were excluded for each assay parameter. The exclusion criteria were: (i) most extreme deviation from the results trend shown by the majority among the 6 experiments, in order to avoid data distortion by outliers (which can be affected e.g. by patient's medication, medical history etc.); and (ii) failure to meet minimal quality criteria.
Administration of Compound A strongly stimulated Keratin-15 immunoreactivity at all concentrations tested (
The aim of the present study is to determine the effect of Compound A on hair shaft elongation.
Details regarding tissue specimen origin used in the present example are as described in Example 2 above.
LDH activity in the supernatant can serve as an indicator of cytotoxicity and was measured following the manufacturer's instructions (Cytotoxicity Detection Kit; Roche, Mannheim, Germany). The absorbance of the samples was measured at 490 nm using an ELISA plate-reader.
Hair shaft length measurements of HFs were performed on individual HFs using a Zeiss inverted binocular microscope with an eyepiece measuring graticule.
HF cycle staging was carried out according to previously defined morphological criteria, and the percentage of HFs in anagen and early, mid, or late catagen was determined.
To evaluate apoptotic cells in colocalization with a proliferation marker Ki-67, a Ki-67/terminal dUTP nick-end labeling (TUNEL) double-staining method was used. Cryostat sections were fixed in paraformaldehyde and ethanol-acetic acid (2:1) and labeled with a digoxigenin-deoxy-UTP (ApopTag fluorescein in situ apoptosis detection kit; Intergen, Purchase, N.Y.) in the presence of terminal deoxynucleotidyl transferase, followed by incubation with a mouse anti-Ki-67 antiserum (1:20 in PBS overnight at 4 C; Dako, Glostrup, Denmark). TUNEL-positive cells were visualized by an antidigoxigenin fluorescein isothiocyanate-conjugated antibody (ApopTag kit), whereas Ki-67 was detected by a rhodamine-labeled goat antimouse antibody (Jackson ImmunoResearch, West Grove, Pa.). Negative controls were performed by omitting terminal deoxynucleotidyltransferase and the Ki-67 antibody. Counterstaining was performed with 4′,6-diamidino-2-phenylindole (DAPI) (Roche Molecular Biochemicals GmbH, Mannheim, Germany). Quantitative immunohistomorphometry was performed; Ki-67-, TUNEL-, or DAPI-positive cells were counted in a previously defined reference region of the HF matrix, and the percentage of Ki-67/TUNEL-positive cells were determined. Statistical analysis was performed using a two-tailed Student's t-test for unpaired samples. For meta-analysis purposes, a total of six assays (each with HFs from a different female individual) were run. In order to avoid data distortion by individual experiments, rigid exclusion criteria were defined that allowed to exclude one individual experiment (out of 5-6) per read-out parameter. Since these exclusion criteria differed for each study parameter, different experiments (i.e. one out of 6) were excluded for each assay parameter. The exclusion criteria were: (i) most extreme deviation from the results trend shown by the majority among the 6 experiments, in order to avoid data distortion by outliers (which can be affected e.g. by patient's medication, medical history etc.); and (ii) failure to meet minimal quality criteria.
Measurement of LDH activity in the supernatant (parameter of cell death and cell lysis) showed a slight increase in LDH activity, only in the 0.1 mM dose on day 6 (
Administration of Compound A slightly, but significantly, inhibited hair shaft elongation in the high dose (1 mM) (
In general, Compound A induced catagen at all concentrations, in agreement with the reduced hair shaft elongation seen with the high dose (1 mM) (
The data indicate that high doses of Compound A (e.g., 1 mM or greater) inhibits hair shaft elongation in occipital human female scalp hair shafts. In addition, all tested concentrations of Compound A have a catagen-inducing effect, which may underlie the decreased hair shaft elongation observed at the high dose.
All publications and patents mentioned herein, including those items listed below, are hereby incorporated by reference in their entirety as if each individual publication or patent was specifically and individually incorporated by reference. In case of conflict, the present application, including any definitions herein, will control.
While specific embodiments of the subject invention have been discussed, the above specification is illustrative and not restrictive. Many variations of the invention will become apparent to those skilled in the art upon review of this specification. The full scope of the invention should be determined by reference to the claims, along with their full scope of equivalents, and the specification, along with such variations.
Unless otherwise indicated, all numbers expressing quantities of ingredients, reaction conditions, and so forth used in the specification and claims are to be understood as being modified in all instances by the term “about.” Accordingly, unless indicated to the contrary, the numerical parameters set forth in this specification and attached claims are approximations that may vary depending upon the desired properties sought to be obtained by the present invention.
The words “comprises/comprising” and the words “having/including” when used herein with reference to the present invention are used to specify the presence of stated features, integers, steps or components but do not preclude the presence or addition of one or more other features, integers, steps, components or groups thereof.
It is appreciated that certain features of the invention, which are, for clarity, described in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the invention which are, for brevity, described in the context of a single embodiment, may also be provided separately or in any suitable sub-combination.
This application claims priority to U.S. Ser. No. 61/700,614, filed Sep. 13, 2012, hereby incorporated by reference in its entirety.
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/EP2013/069063 | 9/13/2013 | WO | 00 |
| Number | Date | Country | |
|---|---|---|---|
| 61700614 | Sep 2012 | US |
