HK1143802B - Compounds with (substituted phenyl)-propenal moiety, their derivatives, biological activity, and uses thereof - Google Patents

Description

Compounds having a (substituted phenyl) -propenal moiety, their derivatives, biological activity and uses thereof

Cross Reference to Related Applications

This application claims priority to U.S. patent application serial No. 60/879,458, filed on 8/1/2007, which is hereby incorporated by reference in its entirety.

Technical Field

The present invention relates to compounds having biological activity and pharmaceutical and cosmetic compositions thereof, their derivatives and methods of use, more particularly, the present invention relates to compounds having at least one (substituted phenyl) -propenal moiety and their biological activity and uses thereof.

Background

It is known that certain natural products will have therapeutic effects that make them useful for the treatment and prevention of human diseases throughout many cultures, such as chinese herbal medicines and many other folk medicines. The effectiveness of such treatments has led the pharmaceutical industry to search for and isolate active substances from these natural products and to develop the active ingredients as therapeutic and prophylactic agents for the treatment and prevention of various diseases or medical conditions. Thus, many of the conventionally used drugs have been developed from or derived from natural products. Including aspirin (acetylsalicylic acid), isolated from the bark of willow; ephedrine and pseudoephedrine, isolated from Ephedra sinica Stapf; and penicillin isolated from fungi (penicillium). However, compounds isolated from natural products are known to exert certain physiological function(s) in their native host; their therapeutic effect on human diseases is not very significant. Historically, such treatments have only been obtained through cumulative experience or "trial and error" in humans. Furthermore, because such compounds were not initially produced for human use, their native form of the compound is often not the optimal form for the treatment of human disease, both structurally and functionally. However, today's advances in modern chemical technology, including analytical and synthetic chemistry, as well as in medical biology, have enabled one to resolve chemical structures and locate "pharmacophores" (core structures essential for therapeutic activity) within a compound, such as one isolated from a natural product; in addition, these new technologies enable one to synthesize new compounds based on pharmacophore structures with optimal or better therapeutic efficacy.

In the present invention, we have demonstrated that compounds having a single (4-hydroxy-3-methoxy-phenyl) -propenal moiety have activity capable of reducing the expression of Androgen Receptor (AR) proteins by enhancing their degradation. This finding was partly derived from our extensive study of ASC-J9(1, 7-bis- (3, 4-dimethoxy-phenyl) -5-hydroxy-hepta-1, 4, 6-triethylenetetramine-3-one), a dimethylated form of the natural compound curcumin (present as the major pigment in turmeric plants). The compound curcumin and many of its isoforms have been reported to have a number of in vitro biological activities, such as antioxidant, anti-inflammatory, anti-tumor and anti-angiogenic activities; however, neither curcumin nor its analogs have yet been developed as therapeutic agents for the treatment of human diseases. This suggests that its natural form of curcumin may not be the optimal molecule for development into a therapeutic drug.

Previously, we have found that the compounds ASC-J9 and ASC-J15 (5-hydroxy-7- (4-hydroxy-3-methoxy-phenyl) -4- [3- (4-hydroxy-3-methoxy-phenyl) -acryloyl ] -hepta-4, 6-dienoic acid ethyl ester) (fig. 1), both possess potent prostate cancer inhibitory and anti-androgenic activity. These two compounds, we have found, also show more potent anti-prostate cancer activity than the current therapeutic drug Hydroxyflutamide (HF), a class of "non-steroidal antiandrogens" widely used in the treatment of human prostate cancer.

After intensive and detailed studies of the structure and biological activity of ASC-J9 and ASC-J15, we have surprisingly found that the (substituted phenyl) -propenal moiety common to both compounds is in fact the core structure attributed to the potent antiandrogen/AR activity of these compounds, and not the entire curcuminoid structure. Based in part on our already completed findings, a number of new compounds, including compounds having one, two, three or four (substituted phenyl) -propenal moieties, further supporting that the (substituted phenyl) -propenal moiety is the pharmacophore of these compounds, are synthesized chemically. Our findings can indicate that an increase in the number of these moieties within the compound structure may alter or increase the anti-androgen/AR activity of the compound. Here we also demonstrate that antiandrogenic activity is present within compounds having a single (substituted phenyl) -propenal moiety. The present inventors have also synthesized new derivatives based on our having at least one (substituted phenyl) -propenal moiety in order to not only elucidate the pharmacophore structure but also to evaluate the antiandrogen and anticancer activity. The novel compounds provided herein by the present inventors further exhibit significant improvements and optimizations in biological activity, bioavailability, water solubility and other criteria necessary for the development of therapeutic drugs.

Disclosure of Invention

The present invention provides biologically active compounds having at least one (substituted phenyl) -propenal moiety. It is therefore an object of the present invention to provide compounds having at least one (substituted phenyl) -propenal moiety for use in the treatment of medical conditions, such as those of the human body.

In one aspect of the present invention, there is provided a compound having at least one (substituted phenyl) -propenal moiety, said compound having the formula of formula I:

wherein 1) R₃And R₄Are each independently selected from the group consisting of alkoxy, hydroxy, and hydrogen; and 2) X is selected fromHydroxyl, alkoxy, ethyl propionate, ethyl methyl carbonate and carbonylalkyl. In some embodiments, the compound has a structure selected from the group consisting of monomers 1, 3, 5, 6, and 7. These monomers are provided as follows:

monomer (b):

monomer 1 monomer 3

Monomer 5 monomer 6

Monomer 7

In another aspect of the invention, there is provided a compound comprising a (substituted phenyl) -propenal moiety having formula (IIa) or (IIb):

wherein, 1) R₃,R₄,R₃' and R₄' is independently selected from the group consisting of-H, -OH and-OCH₃In the group consisting of; 2) l is CO-C8 alkylene or when Z is not, L is unsaturated alkenylene or alkynyl; 3) z is selected from the group consisting of-H, -OH, aromatic ring, cycloalkyl, -COR₁、-CO₂R₁、-CONR₁R₂、-NR₁R₂、-CX₃Wherein R is₁And R₂Independently selected from the group consisting of-H, -CH₃and-C₂H₅And 4) X is a halogen atom selected from the group consisting of-F, -Cl and-Br. Formulas IIa and IIb are equilibrium tautomers which are common phenomena of diketones. In some embodiments, the compound is selected from the group consisting of II-1, II-2, II-3, II-4, and II-5. The formula is provided as follows:

in another aspect of the invention, there is provided a compound of formula IIc:

wherein, 1) R₃、R₄、R₃' and R₄' is independently selected from the group consisting of-H, -OH and-OCH₃In the group consisting of; and 2) R₁And R₂Independently selected from the group consisting of-H, -CH₃、-C₂H₅Substituted aryl and substituted benzyl.

In another aspect of the invention, there is provided a compound of formula III:

wherein R is₃、R₄、R₃’、R₄’R₃"and R₄"is independently selected from the group consisting of alkoxy, hydroxy, and hydrogen. In some embodiments, the compound comprises formula III-1 or III-2, which is provided as follows:

in another aspect of the invention, there is provided a compound of formula IV:

wherein R is₃、R₄、R₃’、R₄′、R₃″、R₄″、R₃' and R₄' are independently selected from the group consisting of alkoxy, hydroxy, and hydrogen. In some embodiments, the compounds include formula IV-I:

in another aspect of the present invention, there is provided a compound of formula V:

wherein, 1) each "n" is independently 1, 2, or 3; 2) r₃、R₄、R₃' and R₄' is independently selected from the group consisting of-H, -OH and-OCH₃In the group consisting of; 3) the L-Z side chain may be absent, but if present, L is C0-C8 alkylene, or when Z is absent, unsaturated alkenylene or alkynyl; 4) z is selected from the group consisting of-H, -OH, aromatic ring, cycloalkyl, -COR₁、-CO₂R₁、-CONR₁R₂、-NR₁R₂、-CX₃In the group consisting of; 5) r₁And R₂Independently selected from the group consisting of-H, -CH₃and-C₂H₅In the group consisting of; and 6) X is selected from the group consisting of-F, -Cl and-BrHalogen atom in (1). In some embodiments, compounds of formula V-I or V-2 are provided:

in another aspect of the invention, a pharmaceutical or cosmetic composition is disclosed comprising a compound having at least one (substituted phenyl) -propenal moiety as provided herein and having a desired biological activity. A pharmaceutical or cosmetic composition may have a compound of the invention and a pharmaceutically or cosmetically acceptable carrier. In various non-limiting embodiments, the compound can include monomer 1, 3, 5, 6, or 7 alone or in combination. In a further embodiment, the compound comprises a formula of formula I, II, III, IV, V or a combination thereof. Thus, the compound may comprise at least one, two, three, four, five or more (substituted phenyl) -propenal moieties.

In yet another aspect of the invention, a method of treating a medical condition is disclosed comprising administering to a person in need thereof a compound containing at least one (substituted phenyl) -propenal moiety and having a desired or suspected biological activity. The compounds may be any one or a combination of those disclosed herein, alone or in combination. The compounds of the invention are useful for treating, preventing or ameliorating symptoms associated with androgen disorders. Non-limiting examples of medical conditions treatable with the disclosed compounds are androgen-associated inflammation including wounds (compounds aid in wound healing), acne, rheumatoid arthritis and alopecia; kennedy' disease; cancers such as prostate cancer, bladder cancer, liver cancer, and breast cancer; and other medical conditions described herein. Treatment of such medical conditions comprises administering to an individual suffering from a medical condition described herein a therapeutically effective amount of any of the disclosed compounds, their derivatives, or pharmaceutical compositions thereof.

Drawings

FIG. 1 shows structural schematic of compounds ASC-J9(1, 7-bis- (3, 4-dimethoxy-phenyl) -5-hydroxy-hepta-1, 4, 6-triethylenetetramine-3-one) and ASC-J15 (5-hydroxy-7- (4-hydroxy-3-methoxy-phenyl) -4- [3- (4-hydroxy-3-methoxy-phenyl) -acryloyl ] -hepta-4, 6-dienoic acid ethyl ester), all of which previously shown to have antiandrogenic activity.

Figure 2 shows a table including a non-limiting list of novel synthetic compounds having at least one (substituted phenyl) -propenal moiety encompassed by the present invention, as well as their structures, chemical formulas and molecular weights.

Figure 3 shows a table of western blot densitometric data demonstrating that compounds with different numbers of (4-hydroxy-3-methoxy-phenyl) -acrolein moieties are capable of reducing Androgen Receptor (AR) protein expression in human prostate cancer CWR22Rvl cells.

Figure 4 shows western blot images demonstrating that the newly provided compounds having at least one (4-hydroxy-3-methoxy-phenyl) -acrolein moiety are capable of reducing Androgen Receptor (AR) protein expression in human prostate cancer CWR22Rvl cells.

Figure 5 shows a table indicating that some selected ASC compounds and monomers are capable of inhibiting proliferation of human prostate cancer cells (LNCaP and CWR22Rvl) under DHT stimulation in vitro.

FIG. 6 shows Western blot data demonstrating that the four compounds ASC-Q49, ASC-Q103, ASC-JM12, and ASC-JM4 are capable of reducing endogenous AR expression in human prostate cancer cells of LNCaP and CWR22Rv1 at different concentrations.

FIG. 7 shows Western blot data demonstrating that compounds ASC-J9 and ASC-JM5 increased degradation of AR protein in the presence of the protein synthesis inhibitor Cycloheximide (CHX) in an assay in LNCaP cells.

Figure 8 shows two tables (8a and 8b) summarizing the efficacy of typical ASC compounds in reducing endogenous AR protein expression (using western blot analysis) at different concentrations when tested in human prostate cancer cells of LNCaP and CWR22Rv 1.

Detailed Description

A. Definition of

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. All patents, applications, published applications and other publications referred to herein are incorporated by reference in their entirety, including the disclosed structures, compositions, methods of use, methods of treatment and methods of manufacture. If there are multiple definitions of terms herein, those in this section prevail unless otherwise indicated.

The term "(substituted phenyl) -acrolein moiety" as used herein means a composition comprising a phenyl group to which an acrolein moiety (when m equals 1) and an alkoxy or hydroxyl moiety, or an alkyl or substituted alkyl moiety, are attached. As used herein, the substitution may be in the meta or para or ortho position with reference to the general formula

Wherein n can be any number of 1, 2, 3 or 4; and m may be a number of 1, 2, 3, 4, or more.

"alkyl" as used herein means a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, containing no unsaturation, having from 1 to 10 carbon atoms, and which is attached to the remainder of the molecule by a single bond, e.g., methyl, ethyl, n-propyl, 1-methylethyl (isopropyl), n-butyl, n-pentyl, 1-dimethylethyl (tert-butyl), and the like.

The term "alkenyl" as used herein means a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, containing at least one double bond, having from 2 to 10 carbon atoms, and which is attached to the remainder of the molecule by a single or double bond, e.g., ethenyl, propenyl, pentenyl, penta-1, 4-dienyl, and the like.

The term "alkenylene" as used herein means a compound containing a carbon-carbon double bond and represented by the formula C_nH_2n-2Straight and branched hydrocarbon chains are represented wherein the hydrogen may be substituted with additional carbon-carbon double bonds or monovalent substituents, such as ethenylene, propenylene, and the like.

The term "alkoxy" as used herein means a group having the formula-OR, wherein R is alkyl, haloalkyl, OR cycloalkyl. "optionally substituted alkoxy" means a group having the formula-OR, wherein R is optionally substituted alkyl as described herein.

The term "alkynyl" as used herein means a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, containing at least one triple bond, having from 2 to 10 carbon atoms, and which is attached to the remainder of the molecule by a single or triple bond, e.g., ethynyl, prop-1-ynyl, but-1-ynyl, pent-3-ynyl, and the like.

The term "aryl" as used herein means a radical of a carbocyclic ring system in which at least one ring is aromatic. The aryl group may be all aromatic rings or may contain aromatic rings in combination with non-aromatic rings. A "bi-aromatic ring system" is a compound comprising at least two aryl groups.

The term "cycloalkyl" as used herein means a monovalent monocyclic or bicyclic hydrocarbon group consisting solely of carbon and hydrogen atoms, having 3 to 10 carbon atoms, which is stable, saturated by single bonds, and attached to the remainder of the molecule, e.g., cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.

The term "diketone bridge" or "ketone-alcohol bridge" as used herein means a straight or branched hydrocarbon chain comprising two ketones or an alcohol located next to a ketone, respectively. A "diketone bridge" or a "ketone-alcohol bridge" is located between at least two aryl moieties.

The term "hydroxyalkyl" as used herein means a straight or branched hydroxy-substituted hydrocarbon chain radical having from 1 to 10 carbon atoms, e.g. -CH₂OH、-(CHz)₂OH and the like.

The term "androgen" as used herein means male hormone hormones such as testosterone and Dihydrotestosterone (DHT). DHT is a product of testosterone transformed by 5-alpha-reductase. Androgens stimulate or control the development and maintenance of male characteristics and other physiological functions in vertebrates and activate or regulate genes by binding to androgen receptors, which in turn bind to androgen/AR-controlled genes (DNA).

The term "androgen receptor" or "AR" as used herein means an intracellular receptor that specifically binds androgens, including testosterone and Dihydrotestosterone (DHT). AR includes all mammalian androgen receptor isomorphs, binding variants and polymorphs.

The term "estrogen receptor" or "ER family" as used herein means an intracellular receptor, particularly for estradiol (the primary endogenous estrogen). When bound to hormones, it acts as a transcription factor (it regulates DNA reading and protein production). ER includes ERA and ER β. ER includes all mammalian isoforms, binding variants and polymorphs of the nuclear receptor.

The term "glucocorticoid receptor" or "GR" as used herein means an intracellular receptor with high affinity for cortisol and other glucocorticoids. GR includes all mammalian isoforms, binding variants and polymorphs of the nuclear receptor.

The term "progesterone receptor" or "PR" as used herein means an intracellular steroid receptor that specifically binds progesterone. PR includes all mammalian isoforms, binding variants and polymorphs of the nuclear receptor.

The term "peroxisome proliferator receptor" or "PPAR" as used herein means all PPAR isoforms, including PPAR α, PPAR β and PPAR γ. The PPAR increases transcription of a target gene by binding to a specific nucleotide sequence in a gene promoter region. When bound to its fatty acid ligand, PPAR α forms a heterodimeric complex with Retinoid X Receptor (RXR) to regulate transcription. PPAR γ is activated by prostaglandins and leukotrienes and regulates gene expression of proteins contained in fatty acid storage. PAR β is slightly activated by fatty acids, prostaglandins, and leukotrienes. The physiological ligand exchange was not recognized.

The term "retinoic acid receptor" or "RAR" as used herein means an intracellular receptor known to bind many retinoid forms. "RAR" includes all family members including RAR α, RAR β and RAR γ. "RAR" includes all mammalian isoforms, binding variants and polymorphs of the nuclear receptor.

The term "retinoid X receptor" or "RXR" as used herein means an intracellular receptor that specifically binds to 9-cis-retinoic acid. "RXR" includes all mammalian isoforms, binding variants and polymorphs of the nuclear receptor.

The term "steroid receptor" or "steroid nuclear receptor" as used herein means an intracellular receptor that binds to DNA and modulates DNA transcription under the regulation of steroid hormones. Receptors for different hormones have strong structural and functional similarities, which apparently evolve from common genetic genes and are therefore considered to be a gene superfamily. Typical receptors belonging to this general family of genes include proteins that are DNA binding and regulated and controlled by the steroid hormones Estradiol (ER), Glucocorticoid (GR), Androgen (AR), Progesterone (PR), Mineralocorticoid (MR), triiodothyronine of the nonsteroidal hormone (T3R) and dihydroxyvitamin D3(VDR), and the two retinoid (all-trans retinoic acid and 9-cis retinoic acid) receptors (RARs and RXRs, respectively). More than 32 genes encoding at least 75 proteins with different DNA specificities, regulators and hormone affinities have been identified as part of this gene superfamily. New members of this general family are frequently and constantly reported and are intended herein to be incorporated by reference in their entirety in parallel published scientific literature or in sequentially provided databases such as gene banks (GenBank) and SWISSPROT, whether DNA, RNA or a polypeptide sequence. Using new biotechnology, molecular biologists and biochemists have identified protein receptors for which ligands have not been identified, thereby creating a class of "orphan receptors". "steroid receptors" include all binding variants and isomers of mammalian steroid receptors.

The term "sustained release" as used herein means a type of administration that provides for a delayed, slow period of time, continuous, intermittent or sustained release of a compound or composition.

By "pharmaceutically acceptable salt" is meant a salt that is approved or approved by a regulatory agency of the federal or a state government for use in animals, and more particularly in humans. The term "pharmaceutically acceptable carrier" means a diluent, adjuvant, excipient, or carrier for approved or approved administration of a compound.

The term "prodrug" as used herein means a compound that is metabolized or otherwise converted, upon in vitro administration, by one or more steps or processes to a biologically, pharmaceutically, or therapeutically active form of the compound. To make prodrugs, the active substance on the drug may be modified in order to regenerate the active compound by metabolic processes. Prodrugs can be designed to alter the stability or transport properties of drug metabolism in order to mask side effects or toxicity, improve drug taste, or alter other properties of drug properties. In some, but not all cases, a prodrug includes a cleavable ester that releases the active form upon cleavage.

The term "therapeutically effective amount" means an amount of a compound that is sufficient to effect treatment of a disease or disorder when the compound is administered to a patient to treat such disease or disorder. The "therapeutically effective amount" will vary depending on the compound, disease or disorder and the severity, age and weight of the patient being treated. A "therapeutically effective amount" may include a range of amounts administered, whether or not the initial amount administered is effective, and ultimately the amount that produces the desired effect.

The term "derivative" as used herein means a variant of the core structure or pharmacophore which produces the desired effect. Derivatives include substitution along the phenyl ring, the acrolein region of the molecule, or along the side chain. Thus, derivatives encompassed herein include compounds formed from or comprising at least one of the disclosed compounds, such as those identified by formula I, II, III, IV, or V. To adjust solubility, efficacy, and polymerizability, etc., it is desirable to form derivatives of specific compounds.

Abbreviations for any protecting groups, amino acids and other compounds used herein are based on accepted abbreviations for their public use or the nomenclature of biochemicals by the IUPAC-IUB Commission (see biochem. 197211: 942-944), unless otherwise specified.

B. Compounds and compositions comprising (substituted phenyl) -propenal moieties

The inventors of the present invention have found that the compounds described herein, including those having at least one (substituted phenyl) -propenal moiety, are promising for the treatment or prevention of medical conditions. In addition, the compounds disclosed herein are believed to have activity, e.g., reduce proliferation of cells believed to have or suspected of having a cancer morphology. In addition, the compounds disclosed herein show the ability to selectively modulate the number of steroid receptors. Accordingly, it is an object of the present invention to provide compounds having biological activity useful for treating or preventing diseases in mammals such as humans.

The present invention discloses and encompasses a variety of compounds and their derivatives for use in the field of medical treatment, such as the treatment or prevention of a medical condition. The compositions disclosed herein may be administered or dosed as the compound per se, or may be adapted with a carrier suitable for the desired therapeutic effect. When the compounds disclosed herein are administered as a medicament, the compounds may be administered in combination with a pharmaceutically acceptable carrier. When the compounds disclosed herein are delivered as a cosmetic, the compounds can be delivered in combination with a cosmetically acceptable carrier. The pharmaceutically acceptable carrier and the cosmetically acceptable carrier may be the same, may be derived from each other known in the pharmaceutical and cosmetic industries, etc., or may be different, e.g., without limitation, depending on the variant of the desired route of administration. The solubility, activity and dipole moment of the compounds before and after preparation as a medicament or cosmetic can be tested and the synergistic effect can be measured alone or in combination with other compounds disclosed herein. Accordingly, the present invention includes one or more compounds and derivatives thereof, including those having hydrophilic or hydrophobic additions, substitutions or subtractions.

In one aspect of the invention, a compound having at least one (substituted phenyl) -propenal moiety is provided. In some embodiments, the compound having a (substituted phenyl) -propenal moiety has biological activity, for example, anti-androgen/anti-AR biological activity. In a particular embodiment of the invention, the (substituted phenyl) -propenal moiety has the formula of formula I:

wherein 1) R₃And R₄Are each independently selected from the group consisting of alkoxy, hydroxy, and hydrogen; and 2) X is selected from the group consisting of hydroxy, alkoxy, ethyl propionate, ethyl methyl carbonate and carbonylalkyl. As can be observed in the figures (fig. s), compounds having at least one (substituted phenyl) -acrolein moiety are capable of reducing androgen receptor or inducing degradation of androgen receptor. Furthermore, compounds having at least one (substituted phenyl) -propenal moiety have been shown to reduce the growth of cancer cells or the proliferation of cancer cells. This inhibition occurs in the presence of a compound that stimulates cancer cells. In various non-limiting embodiments described herein, the compounds include a compound of (substituted phenyl) -propenal, or a pharmaceutically acceptable salt thereof, selected from monomers 1, 3, 5, 6 and 7, alone or in combination. These monomers are provided as follows:

monomer (b):

monomer-1 monomer-3

Monomer-5 monomer-6

Monomer-7

In various embodiments, derivatives of the above-referenced biologically active monomers are also provided. Derivatives may have substituents at one or more positions to increase one or more properties such as activity, solubility, and the like. Such derivatives can modulate the dipole moment of the compound and can form compositions that are more or less hydrophobic or hydrophilic.

In another aspect of the invention, there is provided a compound comprising a (substituted phenyl) -propenal moiety having formula (IIa) or (IIb):

wherein, 1) R₃、R₄、R₃', and R₄' is independently selected from the group consisting of-H, -OH and-OCH₃In the group consisting of; 2) l is CO-C8 alkylene or when Z is not, L is unsaturated alkenylene or alkynyl; 3) z is selected from the group consisting of-H, -OH, aromatic ring, cycloalkyl, -COR₁、-CO₂R₁、-CONR₁R₂、-NR₁R₂、-CX₃Wherein R is₁And R₂Independently selected from the group consisting of-H, -CH₃and-C₂H₅And 4) X is a halogen atom selected from the group consisting of-F, -Cl and-Br. And further wherein formulae IIa and IIb are equilibrium tautomers as a general phenomenon of diketones. In some embodiments, the compound is selected from the group consisting of II-1, II-2, II-3, II-4, and II-5. The formula is provided as follows:

in another aspect of the invention, there is provided a compound of formula IIc:

wherein, 1) R₃、R₄、R₃' and R₄' is independently selected from the group consisting of-H, -OH and-OCH₃In the group consisting of; and 2) R₁And R₂Independently selected from the group consisting of-H, -CH₃、-C₂H₅Substituted aryl and substituted benzyl.

In another aspect of the invention, there is provided a compound of formula III:

wherein R is₃、R₄、R₃’、R₄’R₃"and R₄"is independently selected from the group consisting of alkoxy, hydroxy, and hydrogen. Non-limiting examples include those having the formula III-1 or III-2Those of:

in another aspect of the invention, there is provided a compound of formula IV:

wherein R is₃、R₄、R₃’、R₄′、R₃″、R₄″、R₃' and R₄' are independently selected from the group consisting of alkoxy, hydroxy, and hydrogen. In one embodiment, the compound comprises formula IV-I:

in another aspect of the present invention, there is provided a compound of formula V:

wherein, 1) each "n" is independently 1, 2, or 3; 2) r₃、R₄、R₃' and R₄' is independently selected from the group consisting of-H, -OH and-OCH₃In the group consisting of; 3) the L-Z side chain may be absent, but if the L-Z side chain is present, when Z is absent, L is C0-C8 alkylene or unsaturated alkenylene or alkynyl (alkynl); 4) z is selected from the group consisting of-H, -OH, aromatic ring, cycloalkyl, -COR₁、-CO₂R₁、-CONR₁R₂、-NR₁R₂、-CX₃In the group consisting of; 5) r₁And R₂Independently selected from the group consisting of-H, -CH₃and-C₂H₅In the group consisting of; and 6) X is a halogen atom selected from the group consisting of-F, -Cl and-Br. In some embodiments, the compound has formula V-I or V-2. The following are representative of compounds having the formulas V-1 and V-2:

the compounds of the invention can be synthesized using standard protocols known in the art of organic synthesis using known solvents. The synthesized compounds can be tested for desired activity, such as the ability to degrade steroid receptors such as androgen receptor, prevent or inhibit proliferation of cancer cell lines, and decrease in tumor size in transplanted animal body studies and the like. The use of the synthetic methods and processes disclosed herein are also suitable for compounds identified as being sampled or measured. Thus, the process variations provided will be apparent to those skilled in the art and are considered to fall within the scope of the present invention.

Example 1 illustrates various synthetic schemes for the monomers provided, as well as derivatives thereof, also encompassed herein. In some embodiments, derivatives are provided as combinations of monomers or moieties thereof to form diphenyl, triphenyl or tetraphenyl ring systems or more phenyl ring systems.

In many embodiments, the diphenyl ring system is used in assays and comparisons with other proposed treatments; however, compounds having a single (substituted phenyl) -propenal moiety have also been found to have activity, for example the ability to prevent cancer cell line proliferation under DHT stimulation and the ability to degrade androgen receptors. Some of the compounds of the present invention are prepared by condensing a substituted benzaldehyde with 2, 4-pentanedione or a 3-substituted 2, 4-pentanedione according to methods known in the Pedersen et al, Liebigs Ann. chem., 1557-E.1569 (1985) literature. The desired substitution is synthesized on the diphenyl ring or on C4 of the conjugated bridge, either before or after condensation. The length of the conjugate bridge between the two phenyl moieties can vary from 5 carbon atoms to 11 carbon atoms by synthetic methods. Appropriate addition and removal of protecting groups enables the final synthesis of the disclosed compounds.

Various isoforms and derivatives of compounds having a (substituted phenyl) -propenal moiety have recently been synthesized and evaluated for antiandrogenic activity. Structural information for some, but not all disclosed compounds is summarized in fig. 2.

C. Pharmaceutical and cosmetic compositions containing compounds having at least one (substituted phenyl) -propenal moiety

The invention comprises the disclosed compounds per se, and where appropriate their salts and their prodrugs. The salts or prodrugs should retain a portion of the desired biological activity of the parent compound or be provided in a form in which the body or host can be converted to the biologically active form. For example, a salt can be formed between a positive substituent (e.g., amino group) on a compound and an anion. Suitable anions include, but are not limited to, chloride, bromide, iodide, sulfate, nitrate, phosphate, citrate, methanesulfonate, tartrate, trifluoroacetate, and acetate. Likewise, negative substituents on compounds (e.g., carboxylates) can form salts with cations. Suitable cations include, but are not limited to, sodium, potassium, magnesium, calcium, and ammonium ions such as tetramethylammonium. Non-limiting examples of prodrugs include esters and other pharmaceutically acceptable derivatives, which, when administered to a subject, are capable of providing derivatives of the compounds described above.

The compounds of the present invention may be formulated for administration to prevent and treat a variety of medical conditions. Pharmaceutical compositions can include at least one disclosed compound or a pharmaceutically acceptable salt thereof in combination with a pharmaceutically acceptable carrier. Techniques for the preparation of pharmaceuticals are known in the art and typically involve mixing the compounds or salts in the presence of a suitable carrier. Suitable carriers for use in the compounds of the present invention include diluents, excipients or carrier materials selected for the intended form of administration and to comply with conventional pharmaceutical or cosmetic specifications. Examples of suitable carriers include, but are not limited to, water, physiological saline, phosphate buffered saline, physiologically suitable sustained release agents, saline sustained release with physiologically suitable sustained release agents, water-in-oil and oil-in-water emulsions, alcohols, dimethyl sulfoxide, glucose, mannitol, lactose, glycerol, propylene glycol, polyethylene glycol, polyvinylpyrrolidone, lecithin, albumin, sodium glutamate, cysteine hydrochloride, and the like, and mixtures thereof. Suitable carriers can also include suitable pharmaceutically acceptable antioxidants or reducing agents, preservatives, suspending agents, solubilizers, stabilizers, chelating agents, complexing agents, viscosity modifiers, disintegrants, binders, flavoring agents, colorants, odorants, opacifiers, wetting agents, pH diluents, and mixtures thereof, in accordance with conventional pharmaceutical practice ("Remington: The Science and practice of Pharmacy", 20th edition, Gennaro (ed.) and Gennaro, Lippincott, Williams & Wilkins, 2000).

Depending on the intended route of administration, pharmaceutical and cosmetic compositions may also be provided using methods known in the pharmaceutical and cosmetic arts. Suitable routes of administration may include oral, enteral, parenteral, transmucosal, transdermal, intramuscular, subcutaneous, rectal, intramedullary, intrathecal, intravenous, intraventricular, intra-atrial, intra-aortic, intra-arterial, or intraperitoneal administration.

The pharmaceutical compositions of the present invention may be administered to a subject via medical devices such as, but not limited to, implantable devices, biodegradable implants, patches and pumps. In the case of such devices, compositions containing a decomposable or non-decomposable matrix or medium (e.g., a coating, film, infiltrated matrix, polymer, sponge, gel, or porous layer on or within a medical device) can be formulated to enable release of the active compound over a specified period of time.

For use in living, whole organisms, such as humans, the compositions of the invention can be formulated in any composition suitable for the intended administration form and in accordance with conventional pharmaceutical practice andsupply ("Remington: The Science and Practice of Pharmacy", 20)^th edition，Gennaro(ed.)and Gennaro，Lippincott，Williams &Wilkins, 2000). Examples of suitable compositions include tablets, capsules, syrups, elixirs, ointments, creams, lotions, sprays, aerosols, inhalants, solid particles, powders, microparticles, gels, suppositories, concentrates, emulsions, liposomes, microspheres, dissolvable matrices, sterile liquids, suspensions or injections, and the like. Injectables can be prepared in conventional forms, either as liquid solutions or suspensions, as concentrates or solid particles suitable for solution or liquid suspension prior to injection, or as emulsions.

D. Medical treatment incorporating compounds having at least one (substituted phenyl) -propenal moiety

The compounds of the invention were tested for their effect on steroid receptors and their effect on cancer cell numbers. As a result, it was found that the compound of the present invention was able to reduce the expression of androgen receptor (see fig. 3 and 4). Further studies have shown that the compounds of the present invention are capable of inhibiting the growth of cancer cells (see figure 5) and reducing the expression of androgen receptors in cancer cells (see figures 6 and 8). The inventors also believe the underlying mechanism of action or underlying pathway. Figure 7 supports the inventors' view that the compounds of the present invention induce degradation of the androgen receptor. Thus, the activities shown herein support the therapeutic or prophylactic treatment of medical conditions such as various cancers and androgen-related disorders.

The present invention provides methods of treating, ameliorating, or preventing the development of symptoms of various medical conditions using the compounds and compositions of the present invention, including pharmaceutical and cosmetic compositions. The medical condition may be modulated, at least in part, by steroid receptors. Steroid receptors of particular interest may include, but are not limited to, Androgen Receptor (AR), Progesterone Receptor (PR), Estrogen Receptor (ER), Glucocorticoid Receptor (GR), peroxisome proliferator-activated receptor (PPAR), retinoic acid receptor (RARs and RXRs), and orphan steroid hormone receptors. The compositions or compounds of the present invention may target a specific receptor, such as the androgen receptor, or may target a specific receptor within the steroid receptor superfamily.

The methods of the invention can prevent, treat or ameliorate the symptoms of cancer such as, but not limited to, prostate cancer, liver cancer, bladder cancer, cervical cancer, lung and breast cancer, skin cancer, small cell lung cancer, testicular cancer, lymphoma, leukemia, esophageal cancer, stomach cancer, colon cancer, endometrial cancer, ovarian cancer, central nervous system cancer, and the like. The methods of the invention can induce cytotoxicity of anti-tumor cells or can inhibit growth of tumor cells. Determining whether a compound or drug is beneficial for treating or preventing a particular disease may include testing the compound or their derivatives in vitro, in vivo in animal models or in cell-based assays on appropriate cell lines, in the case of cancer, cell lines having the morphology of the cancer cells, such as cell lines prepared from cancer cells, may be used. In some embodiments, the compounds of the invention are evaluated for their activity in inhibiting the growth or proliferation of cancer cells, optionally stimulated with a stimulating agent such as DHT. The compounds disclosed herein are particularly shown to reduce the growth or proliferation of prostate cancer cells.

In other embodiments, the compounds, their derivatives, pharmaceutical compositions and the like are useful for preventing, treating or ameliorating neurological and neuromuscular disorders such as Kennedy's disease (Kennedy's disease). Spinal and Bulbar Muscular Atrophy (SBMA) or kennedy's disease is a sex-specific motor neuropathy that infects 1 in 40,000 men (reviews by Katsuno et al, 2004). SBMA patients have a variant androgen receptor that contains an expanded polyglutamine tract. The expanded polyglutamine androgen receptor forms aggregates which impede the functioning of the cell and are the cause of adjacent muscle atrophy associated with SBMA. The method of the invention comprises mitigating aggregate-induced stress by reducing the number of mutant AR to a level that can be more easily regulated by a cellular parental finishing instrument. The methods of the invention may include selectively degrading the androgen receptor and thus may be useful in the treatment of SBMA. Compounds of the invention that are capable of increasing androgen receptor degradation may inhibit the steady state level of the receptor, thereby reducing the extent of aggregate formation in the patient.

The compounds and compositions of the present invention can prevent, treat or ameliorate the symptoms of androgen-associated hair disorders. For example, androgenic alopecia or "male pattern baldness" is the loss of hair caused by the action of androgen activity on androgen receptors in follicles and neighboring cells. As another example, hirsutism refers to the excessive growth of dark hair in areas where women generally have little or no hair growth. This eventual male growth of body hair usually occurs at androgen-stimulated sites such as the face, chest and breast. The methods of the invention may comprise administering a compound, pharmaceutical or cosmetic composition to a subject in need of such treatment or prevention.

The compounds and compositions of the present invention can treat inflammation (e.g., rheumatoid arthritis), acne, alopecia, and can accelerate wound healing. Acne is caused by androgen-induced AR activation of the sebaceous glands and can therefore be treated by administering compounds capable of preventing or reducing AR activation. The compounds of the present invention are believed to induce degradation of the androgen receptor and therefore would provide an effective treatment for this medical condition. Androgenic alopecia and other hair growth disorders are known to be caused by endogenous androgen activation of Androgen Receptors (ARs) in hair follicles. Certain inflammatory states and wound healing are also thought to be associated with androgen receptors that respond to androgens. The methods of the present invention comprise administering a compound, pharmaceutical or cosmetic composition to an individual in need of such treatment or prevention. Topical application of such compositions may be particularly important.

The compounds and compositions of the invention are useful in the treatment of endocrine disorders. Androgen excess is one of the most common endocrine disorders in women (review by Bulun and Adashi, 2003). This pathophysiological condition can be found in women with different endocrine disorders, including polycystic ovary syndrome (PCOS), pituitary adenoma-induced hyperprolactinemia, cushing's syndrome, congenital adrenal hyperplasia, atypical adrenal hyperplasia, ovarian or adrenal neoplasia, and iatrogenic androgen excess. Of these disorders, PCOS, which occurs in 5% to 10% of women of childbearing age, is the most frequently established cause of hyperandrogenism. Recently, a relative increase in the ratio of circulating androgens to circulating estrogens (termed androgenic performance) has been observed in postmenopausal women (Lee et al, 2004). Androgenic performance is the result of a greater reduction in the synthesis of estradiol and estrone than androgen synthesis after menopause, and its clinical use is under active investigation. It has been shown that women with androgen excess are more often obese in vivo (Peohlman et al, 1995). The fat deposited in the abdominal wall is metabolically active and is associated with insulin resistance in the surrounding tissues (Evans et al, 1983). In addition to the endocrine disorders described above, symptoms of excess androgens can also be detected in AIDS (HIV) -infected women who exhibit symptoms of lipodystrophy (Hadigan et al, 2000). It has been suggested that androgenicity may involve lipid variants observed in the latter group of patients.

The methods of the invention include treating the various medical conditions disclosed herein, or are believed to be at least partially associated with steroids or steroid-related disorders. The method of treatment comprises administering a compound, pharmaceutical composition or cosmetic composition of the invention to an individual or subject in need thereof. The subject may be treated with a therapeutically effective amount of the dose. The therapeutically effective dose may vary somewhat from compound to compound, patient to patient and depending on the condition of the patient and the route of delivery. As a general guide, dosages of from about 0.1 to about 50mg/kg have therapeutic efficacy, while higher dosages may also be used.

Many features of the present invention will be described in more detail in the following non-limiting examples. The following examples are therefore provided to further illustrate various aspects and embodiments of the present invention. It is to be understood, however, that the invention as described herein, and as recited in the claims, is not intended to be limited by the details of the following examples.

Examples

Example 1: preparation of compounds and derivatives having at least one (3, 4-alkoxy or hydroxy-substituted phenyl) -propenal moiety

In some embodiments, compounds consisting of a single (substituted phenyl) -acrolein core building block (monomer) are prepared by standard and advanced organic synthesis. In some embodiments, compounds consisting of two or more (substituted phenyl) -acrolein core moieties are prepared by polycondensation of substituted benzaldehydes and 2, 4-pentanediones or 3-substituted 2, 4-pentanediones using methods known in Pedersen et al, Inc. (Liebigs Ann. chem., 1557-. The desired substituents on the diphenyl and on the C4 of the conjugated bridge are synthesized either before or after polycondensation. The length of the conjugate bridge between the two phenyl moieties can vary from 5 carbon atoms to 11 carbon atoms by synthetic methods. Appropriate addition and removal of protecting groups enables the final synthesis of the derivatives of the invention. Furthermore, in order to obtain the desired compound, a plurality of synthetic steps may be performed in sequence.

The derivatized phosphate prodrugs are further prepared by reacting a compound having a (substituted phenyl) -propenal moiety with a phosphorus-containing oxychloride in the presence of an organic base such as triethylamine in a suitable solvent such as dichloromethane. The tartrate salt of the compound of the present invention as a water-soluble salt can be prepared by reacting a compound having a (substituted phenyl) -acrolein moiety with tartaric acid in water.

Chemical synthesis

Melting points were determined on a Fisher-John melting point apparatus and were not corrected. Proton nuclear magnetic resonance (¹HNMR) and¹³the C NMR spectra were measured on a Varian Gemini 300 or Inova 500 spectrometer with Tetramethylsilane (TMS) as an internal standard. Determination on a 500MHz Varian Inova spectrometer using phosphoric acid as external standard³¹P NMR. The isomeric shift is reported as δ (ppm). Mass Spectrometry (MS) at Agilent1100 series LC-MSD-Trap or PE-Sciex API-3000 spectrometer. Flash column chromatography was performed on silica gel (100-. HPLC was performed on Shimadzu SCL 1OA equipment. HPFC is carried out on a Biotage system or on a Chemflash chromatography system from ISCO. Preparative Thin Layer Chromatography (PTLC) on silica gel plates (Kieselgel 60, F254, 0.25mm) was also used for separation and purification. Precoated silica gel plates (Kieselgel 60, F254, 0.25mm) were used for analysis by Thin Layer Chromatography (TLC). ASC-J9, a starting material, was synthesized by reacting 3, 4-dimethoxybenzaldehyde with 2, 4-pentanedione according to a published method (Pedersen et al, Liebigs Ann. chem., 1557-H-E1569, 1985).

Synthesis of monomers 1, 3, 5-7

The basic structural monomers, which have a (3, 4-dimethoxy or 3-methoxy-4, hydroxy-substituted phenyl) -propenal moiety in their structure and which now provide compounds, have been synthesized by the reaction of 3- (3 ', 4' -dimethoxyphenyl) -propenoic acid with the corresponding reagent (monomer 1, 3) or the reaction of 3, 4-dimethoxybenzaldehyde with 3-methoxy-4-p-hydroxybenzaldehyde with ethyl levulinate (monomers 5 and 6). Monomer-7 was synthesized from 3- (3, 4-dimethoxyphenyl) propane starting in two steps. More specifically, the method of synthesis of the monomers is described below and illustrated in scheme 1.

Monomer 1, methyl 3- (3 ', 4' -dimethoxy-phenyl) -acrylate, was synthesized by reacting 3- (3 ', 4' -dimethoxy-phenyl) -acrylic acid with methanol in the presence of acetyl chloride. After 2.5h of reflux, the reaction mixture was concentrated by evaporation to 1/3, the white solid was filtered and dried in vacuo to give the desired product as a white crystalline solid in 76% yield, mp.74-75 ℃.

ESI MS m/z：223.0[M+H]⁺；¹H NMR(300MHz，CDCl₃)δ：7.64(d，IH，J＝15.9Hz，H-3)，7.11(dd，IH，J＝6.9，2.1Hz，H-6′)，7.05(d，IH，J＝2.4Hz，H-2′)，6.85(d，IH，J＝8.4Hz，H-5′)，6.32(d，IH，J＝15.9Hz，H-2)，3.92(s，6HPhenyl OCH₃) 3.80(s, 3H, ester OCH)₃)。

Monomer 3, a mixed anhydride, by reaction of 3- (3 ', 4' -dimethoxy-phenyl) -acrylic acid in toluene/CH₂Cl₂In Et (1: 1)₃In the presence of N. The solution was cooled to 0 ℃ and ethyl chloroformate (1.5eq.) was added dropwise. After stirring for 2h at 0 ℃ the precipitate was filtered off. The filtrate was concentrated to give a relatively pure white solid, purified by permanent filtration through a thin pad of silica gel and eluted with hexane: ethyl acetate (1: 0-4: 1) to give the desired product as a large white solid. ESI MS m/z: 281.0[ M + H]⁺；¹H NMR(300MHz，CDCl₃)δ：7.78(d，IH，J＝15.9Hz，H-3)，7.15(dd，IH，J＝8.4，1.8Hz，H-6′)，7.06(d，IH，J＝1.8Hz，H-2′)，6.89(d，IH，J＝8.4Hz，H-5′)，6.29(d，IH，J＝15.9Hz，H-2)，4.37(q，2H，J＝6.9Hz，OCH₂OB), 3.92(d, 6H, J ═ 1.2Hz, phenyl OCH)₃)，1.40(t，3H，J＝7.2，OCH₂CH₃).

Monomer 5, namely 6- (3 ', 4' -dimethoxy-phenyl) -4-oxo-hexa-5-enoic acid ethyl ester, was synthesized by reacting 3, 4-dimethoxy-benzaldehyde with ethyl levulinate, as shown in scheme 1. Ethyl levulinate (1eq.) was reacted with boron oxide (0.7eq) in ethyl acetate at 40 ℃ for 30 min. To the obtained solution were added tributyl borate and 3, 4-dimethyloxybenzaldehyde (both 1eq.) and the solution was stirred at 40-42 ℃ for 30 minutes. Butylamine (0.7eq) ethyl acetate solution was added slowly and the mixture was stirred at 40-42 ℃ overnight. Hydrochloric acid (1.3eq) was added and stirred at 60 ℃ for 1 hour. The reaction mixture was cooled to room temperature and separated. The aqueous phase was extracted 2 times with ethyl acetate. The combined ethyl acetate extracts were washed with water to pH 4 and MgSO₄And (5) drying. After filtration and concentration, the precipitate was purified by PTLC to afford monomer 5 as a white solid. mp.62-63 ℃ ESI MS m/z: 293.2[ M + H]⁺；¹H NMR(300MHz，CDCl₃)δ：7.55(d，IH，J＝16.2Hz，H-6)，7.14(dd，IH，J＝9.0，2.1Hz，H-6′)，7.08(d，IH，J＝1.8Hz，H-2′)，6.88(d，IH，J＝8.4Hz，H-5′)，6.65(d，IH，J＝16.2Hz，H-5)，4.16(q，2H，J＝6.9Hz，OCH₂CH₃) 3.93(s, 6H, phenyl OCH)₃)，3.01(t，2H，J＝6.6Hz，H-3)，2.69(t，2H，J＝6.6Hz，H-2)，1.27(t，3H，J＝6.9，OCH₂CH₁)。

The monomer 6, 6- (4-hydroxy-3-methoxy-phenyl) -4-oxo-hexa-5-enoic acid ethyl ester was synthesized by reacting vanillin with ethyl levulinate in the same method as recorded in the synthesis of monomer 5. The desired compound was obtained as a yellow crystalline solid. ESI MS m/z: 279.2[ M + H]⁺；¹H NMR(300MHZ，CDCl₃) δ: 7.54(d, IH, J ═ 15.0Hz, H-6), 7.12-7.06(m, 2H, aromatic-H), 6.94(d, IH, J ═ 8.1Hz, aromatic H-5'), 6.63(d, IH, J ═ 15.0Hz, H-5), 4.16(q, 2H, J ═ 7.2Hz, OCH₂CH₃) 3.94(s, 3H, phenyl OCH)₃)，3.01(t，2H，J＝6.9Hz，H-3)，2.69(t，2H，J＝6.9Hz，H-2)，1.27(t，3H，J＝7.2，OCH₂CH₁)。

The monomer 7, i.e. 7- (3, 4-dimethoxy-phenyl) -hept-6-ene-2, 5-dione, was prepared in two steps from 3- (3, 4-dimethoxyphenyl) propane. 3, 4-Dimethoxycinnamaldehyde was prepared in 60% yield as described for the synthesis of Q110 (scheme 13). The obtained compound (1eq.) was dissolved in anhydrous EtOH and 3-buten-2-one was added. The reaction solution was heated to 80 ℃ under N2, and a solution of 3-benzyl-5- (2-hydroxyethyl) -4-methyl-1, 3-thiazolium chloride (0.1eq.) and TEA (0.4eq.) in EtOH was added dropwise. The resulting reaction mixture was stirred at this temperature for 10 hours and then evaporated to give a yellow precipitate. Dissolving the precipitate in CH₂Cl₂In combination with 0.5% H₂SO₄、2％NaHCO₃And a saline flush. Through Na₂SO₄Drying, and passing through Al by chromatography₂O₃The precipitate was purified by flash column and subsequently crystallized from ether and pentane to obtain the title compound as an off-white solid, mp.71-73 ℃ ESI MS m/z: 263.0[ M + H]⁺；¹H NMR(300MHz，CDCl₃)δ：7.55(d，IH，J＝16.2Hz，H-6)，7.14(dd，IH，J＝9.9，2.1Hz，H-6′)，7.08(d，IH₅J ═ 1.8Hz, H-2 '), 6.88(d, IH, J ═ 8.4Hz, H-5'), 6.64(d, IH, J ═ 16.2Hz, H-5), 3.93(s, 6H, phenyl OCH)₃)，2.98(t，2H，J＝6.0Hz，H-3)，2.83(t，2H，J＝6.0Hz，H-3)，2.24(s，3H，COCH₃)。

Scheme 1

1. Synthesis of monomer 1:

2. synthesis of monomer 3:

3. synthesis of monomer 5:

4. synthesis of monomer 6:

5. synthesis of monomer 7:

catalyst: 3-benzyl-5- (2-hydroxyethyl) -4-methyl-1, 3-thiazolium chloride monomer 7

Synthesis of Compounds Q9, Q44, Q49, Q50, Q77 and Q98

To investigate the effect of C4-substitution of compounds on AR activity, C4-substituted compounds with different functional groups (e.g., hydroxy, ester, amide, etc.) were synthesized. These compounds are prepared by treating 1, 7-bis- (3, 4-dimethoxy-phenyl) -5-hydroxy-hept-1, 4, 6-triethylenetetramine-3-one (ASC-J9), synthesized by the method described in scheme 2, with the appropriate bromide or chloride compound (or ethylene oxide as an alternative for making compound Q9) under basic conditions.

Compound Q9 was synthesized as follows. To an aqueous 1N NaOH solution containing 0.1mmol of tetrabutylammonium bromide (phase transfer catalyst, PTC) was added CH₂Cl₂ASC-J9(0.1mmol) in (0.5 mL). The mixture was stirred at room temperature for 10 minutes and 2-bromoethanol alcohol (0.2mmol) or ethylene oxide (25mmol) was added. The resulting mixture was stirred at 40 ℃ overnight for the Q9 compound. Separating the two layers and using CH₂Cl₂The aqueous phase was extracted 3 times. Through Na₂SO₄The combined organic layers were dried and concentrated. The crude residue was purified by PTLC and crystallized from EtOAc. Analytical data for compound Q9 are shown below.

Compound Q9: yellow crystalline solid (EtOAc), mp.149-150 ℃ ESI MS m/z: 441.3[ M + H]⁺；¹H NMR (300MHz, CDCl3) delta: 7.63(d, IH, J ═ 15.9Hz, H-I), 7.53(d, IH, J ═ 15.9Hz, H-7), 7.14-7.04(m, 4H, aromatic ring H), 6.88-6.85(2H, aromatic ring H), 6.65(d, IH, J ═ 15.9Hz, H-2), 6.31(d, IH, J ═ 15.9Hz, H-6), 4.29(t, 2H, J ═ 12, and 6Hz, CH ═ 15.9Hz, H-6)₂CH₂OH)，3.94-3.88(12H，OCH₃，)，2.84-2.79(t，IH，C4-H)，2.14-2.10(m，2H，CH₁CH₂OH)。

Compounds Q44, Q49, Q77 are prepared by reacting at K₂CO₃And Cs₂CO₃(9: 1) or in the presence of NaH₂Cl₂Or ASC-J9 in THF with the appropriate bromide or chloride compound as shown in scheme 2. For the example of making compounds Q49 and Q77, ASC-J9(1eq) was added to a solution of NaH (4eq.) in THF at 0 ℃. The resulting solution was stirred at 0 ℃ for 0.5h, then at room temperature for 1.5 h. 2-chloride-NN-diethylacetamide (4eq.) (for Q49) or 2-chloride-NN-dimethylacetamide (4eq.) (for Q77) was added. The resulting mixture was heated to reflux overnight. The reaction mixture was cooled to room temperature (r.t.), diluted with EtOAc and 10% H₂SO₄And (4) cleaning with the solution. With saturated NaHCO₃，H₂The organic layer was further washed with O and brine and Na₂SO₄And (5) drying. The desired product was purified by flash column chromatography and crystallized from EtOAc.

Compound Q49: yellow crystalline solid, mp.166-167 ℃. ESI MS m/z: 510.7[ M + H]⁺；¹H NMR (300MHz, CDCl3) delta: 7.68(d, 2H, J ═ 15.9Hz, H-1, 7), 7.16-7.06(4H, aromatic ring H), 6.87-6.84(2H, aromatic ring H), 6.80(d, 2H, J ═ 15.9Hz, H-2, 6), 4.97(t, IH, J ═ 12.0 and 6.0Hz, C4-H), 3.92-3.89(12H, OCH, H₃)，3.43-3.33(m，4H，CH₂CON(CH₂.CH₃)₂)，3.04(d，2H，J＝6.6Hz，C4-CH₂CON(CH₂CH₃)₂)，1.24(t，3H，CH₂CON(CH₂CH₃J)₂)，1.09(t，3H，CH₂CON(CH₂CH₃)₂)。

Compound Q77: yellow crystalline solid, rap.155-157 ℃ ESI MS m/z: 482.2[ M + H ]]⁺；¹H NMR (300MHz, CDCl3) delta: 7.68(d, 2H, J ═ 15.6Hz, H-1, 7), 7.16-7.06(4H, aromatic ring H), 6.87-6.83(2H, aromatic ring H), 6.77(d, 2H, J ═ 15.6Hz, H-2, 6), 4.92(t, IH, J ═ 13.5 and 6.6Hz, C4-H), 3.92-3.88(12H, OCH, H₃)，3.09-3.04(m，5H，-CH₂COand N(CH₂))，2.94(s，3H，N(CH₃))。

Compounds Q50 and Q98 were synthesized to compare their activities with Q44 and Q49 (scheme 3). To 5-hydroxy-1, 7-bis- (4-hydroxy-3-methoxy-phenyl) -hepta-1, 4, 6-triethylenetetraminePyridine chlorochromate (PPTS) (0.1eq.) was added to a solution of (E) -3-ketone and 3, 4-dihydro-2H-pyran (20 eq.) in anhydrous dichloromethane. The resulting solution was stirred at room temperature for 48 h. The solution was then washed with water. The solvent was removed and the obtained compound was purified on a Biotage column chromatograph. The product obtained (Q1) was reacted with ethyl bromoacetate (Q50) or 2-chloride-N, N-diethylacetamide (4eq.) (Q98) at K₂CO₃And Cs₂CO₃(9: 1) reaction in the presence of PPTS/EtOc followed by removal of the Tp protecting group afforded the strong products Q50 and Q98, respectively.

Compound Q50: amorphous state, mp.63-65 ℃ ESI MS m/z: 455.2[ M + eta ]]⁺；¹H NMR (300MHz, CDCl3) delta: 7.65(d, 2H, J ═ 15.9Hz, H-1, 7), 7.19-7.04(6H, aromatic ring H), 6.72(d, 2H, J ═ 15.9Hz, H-2, 6), 4.16(2H, COOCH₂CH₃)，3.96-3.92(6H，OCH₃)，3.04(d，2H，J＝7.2Hz，C4-CH₂COOCH₂CH₃)，1.27-1.23(3H，COOCH₂CH₃)。

Compound Q98: amorphous state, mp.68-71 ℃ ESI MS m/z: 482.10[ M + H]⁺；¹HNMR (300MHz, CDCl3) δ: 7.65(d, 2H, J ═ 15.9Hz, H-1, 7), 7.12-7.03(4H, aromatic ring H), 6.94-6.89(2H, aromatic ring H), 6.76(d, 2H, J ═ 15.9Hz, H-2, 6), 4.96(t, IH, J ═ 13.2 and 6.9Hz, C4-H), 3.92-3.89(6H, OCH, H₃)，3.44-3.33(m，4H，CH₂CON(CH₂CH₃)，)，3.04(d，2H，J＝6.6Hz，C4-CH₂CON(CH₂CH₃)₂)，1.25(t，3H，CH₂CON(CH₂CH₃)₂)，1.10(t，3H，CH₂CON(CH₂CH₃J₂)。

Scheme 2

Compound Q9

Compound Q44: n ═ 1, R ═ COOC₂H₅

Compound Q49: n is 1, R is CON (C)₂H₅)₂

Compound Q77: n is 1, R is CON (CH)₃)₂

Scheme 3

5-hydroxy-1, 7-bis- (4-hydroxy-3-methoxy-phenyl) -hepta-1, 4, 6-triethylenetetramine-3-one

Synthesis of the Compounds: q12

This compound was initially synthesized from commercially available substituted benzaldehydes and 4-acetyl-5-oxohexanoate as shown in scheme 4.

More specifically, 4-acetyl-5-oxohexanoate salt was reacted with boron oxide (0.7eq) in ethyl acetate at 40 ℃ for 30 minutes. To the obtained mixture were added tributyl borate and 3-methyl-4-p-hydroxybenzaldehyde (both 1.6 to 1.8eq) and the mixture was stirred at 40 to 42 ℃ for 30 minutes. Butylamine (1.5eq) Ethyl Ether was added slowlyThe ethyl ester solution was acidified and the mixture was stirred at 40-42 ℃ overnight. 10% hydrochloric acid (2.5eq) was added and stirred at 60 ℃ for 1 hour. The reaction mixture was cooled to room temperature and separated. The aqueous phase was extracted 2 times with ethyl acetate. The combined ethyl acetate extracts were washed with water to pH 4 and MgSO₄And (5) drying. After filtration and concentration, the precipitate was purified by chromatography on silica gel using hexane: ethyl acetate as eluent and crystallized from ethyl acetate.

Scheme 4

Synthesis of the Compounds: q30, Q35 and Q70

To investigate the function of the diketone group in AR activity, a series of compounds with an imine group instead of one of the ketones were synthesized.

Compounds Q30, Q35 by reacting at BF₃·OEt₂Synthesized in the presence of ASC-J9 by reaction with the appropriate amine (scheme 5). For example, 7-diethylamine (compound Q30) (1.2eq.) was added to a solution of ASC-J9 in 1, 2-dichloroethane. The solution obtained is cooled to-30 ℃ and fresh BF is added dropwise₃·OEt₂(2 eq.). The mixture was stirred at-30 ℃ to room temperature under nitrogen and monitored by TLC. After cooling by addition of pyridine (ca. 3eq.), the mixture is washed with brine and over MgSO₄And (5) drying. Evaporation of the solvent and purification by flash column chromatography gave the desired product Q30, ESI MS m/z: 452.4[ M + H]⁺。

Compound Q70 was synthesized by reacting ASC-J9 and (0.75mmol) with (R) - (-) -2-phenylglycinol (1.16mmol) in dry toluene as shown in scheme 5. The reaction mixture was heated to reflux with a Dean-Stark trap (Dean-Stark trap) overnight. The solvent was evaporated and ethyl acetate was added and evaporated again. The obtained precipitate was purified by column chromatography on Biotage system to obtain the desired product as a light yellow solid, ESI MS m/z: 516.4[M+H]⁺；¹H NMR (300MHz, CDCl3) delta: 7.54(d, IH, J-15.6Hz, H-I), 7.40-7.28(5H, aromatic ring H), 7.13(d, IH, J ═ 15.9Hz, H-6), 7.16-7.09(2H, aromatic ring H), 6.95-6.80(4H, aromatic ring H), 6.68(d, IH, J ═ 15.6Hz, H-2), 6.63(d, IH, J ═ 15.9Hz, H-7), 5.63(s, IH, C4-H), 3.94-3.83(m, 15H).

Scheme 5

Compound Q35, R ═ CH (CH)₂)2CH₃

Compound Q70

Synthesis of the Compounds: synthesis of Q99, Q106, Q113, JM2 and JM20

In an ongoing study of the effect of the C4 side chain on AR activity, a series of C4 substituent-containing compounds with carboxyl groups were synthesized. Compound Q99 was synthesized by reaction of ASCJ-9 with 3-chloro-2-methoxymethoxy-propene followed by removal of the methoxymethyl group as shown in scheme 6. More specifically, an aqueous solution of NaOH (2eq.) and tetrabutylammonium hydrogen sulfate (TBABS) were stirred for 5 minutes. To the reaction solution was added dropwise a solution of ASCJ-9(1eq.) in 1, 4-dioxane at room temperature and the obtained red two-phase mixture was stirred at room temperature for 10 minutes. To the mixture was added a solution of 3-chloro-2-methoxymethyloxy-propene (1.5eq.) in 1, 4-dioxane and the obtained solution was stirred at room temperature for 5 minutes and then at 70 ℃ overnight. The solid was removed by filtration and the filtrate was concentrated to dryness. The residue obtained is suspended in 1% H₂SO₄In dioxane (2: 1, volume ratio) and the suspension is stirred at room temperature for 4 hours under TLC monitoring. Mixing the reaction mixtureBy CH₂Cl₂Extracting with Na₂SO₄Dried, filtered and concentrated. The precipitate was purified by flash column chromatography and eluted with a hexane/EtOAc mixture to give the desired product as a yellow crystalline solid. M.p.163-166 ℃, ESI MS m/z: 453.1[ M + H]⁺。

Scheme 6

Compound Q99

Compounds Q106 and Q113 were synthesized using the method described above for the production of compound Q44 in scheme 2. An example of fabricating Q106 is as follows. To ASC-J9(0.25mmol) of anhydrous CH₂Cl₂Adding bromo-1-phenyl ethyl ketone (1.2eq.), K into the (5rnL) solution₂CO₃/Cs₂CO₃(10: 1) (-2 eq.). The reaction mixture was stirred at room temperature overnight, monitored by TLC. The reaction mixture was diluted with EtOAc and washed with water, then Na₂SO₄And (5) drying. The crude product obtained was purified by flash column chromatography on silica gel eluting with a mixture of hexane and EtOAc to give the desired product.

Compound Q106, yellow crystals, mp.160-2 ℃ ESI MS m/z: 515.2[ M + H]⁺.¹HNMR(300MHz，CDCl₃) δ: 8.14-8.01(2H, aromatic ring H), 7.74-7.68(2H, H-1, 7), 7.65-7.46, (m, 4H, aromatic ring H), 7.18-7.15(IH, aromatic ring H), 7.09-7.06(2H, aromatic ring H), 6.91-6.80(m, 4H, aromatic ring H), 6.69(d, 2H, J-15.3Hz, H-2, 6), 3.92-3.90(12H, OCH)₃)，3.78(2H，-CH₂CO)。

Compound Q113, yellow fluffy solid, mp.145-7 ℃ ESI MS m/z: 479.1[ M + H ]]⁺.¹HNMR(300MHz，CDCl₃) δ: 7.75(d, IH, J ═ 15.3Hz, H-1, 7), 7.20-7.17(2H, aromatic ring H), 7.07-7.06(2H, aromatic ring H), 6.90, 6.88(2H, aromatic ring H), 6.86(IH, J ═ 15.3Hz, H-2，6)，3.95-3.93(12H，OCH₃)，3.76(2H，-CH₂CO), 2.14-2.05(m, IH, cyclopropyl-H), 1.10-1.04(m, 2H, cyclopropyl-H), 0.93-0.86(m, 2H, cyclopropyl-H).

Compound JM was synthesized by reaction of ASC-J9(40mg) with iodoacetamide (80mg) and anhydrous sodium carbonate (40mg) in anhydrous acetone as shown in scheme 7. The reaction mixture was heated to reflux for 24 hours. After cooling, the mixture was filtered to remove inorganic solids and the filtrate was evaporated. The crude residue obtained was purified by preparative silica gel chromatography plate (ethyl acetate only) to obtain the desired product as a light yellow solid.

Compound JM2, amorphous; ESI MS m/z: 452.2[ M + H]⁺；¹H NMR (300MHz, CDCl3) delta: 7.79(d, 2H, J-15.3Hz, H-I, 7), 7.4-6.4(6H, aromatic ring H), 6.33(d, 2H, J ═ 15.3Hz, H-2, 6), 3.93, 3.92 (all s, two 6H, OCH)₃)，2.06(d，J＝6.3Hz，2H，CH₂CONH₂)。

Synthesis of JM-IO A solution of 1.0g of ASC-J9, 5ml of acetic anhydride, 1ml of trimethyl orthoformate was stirred at 70 ℃ for 22 hours (scheme 7). The solution was then evaporated to dryness in vacuo. The residue was redissolved in CH₂Cl₂-ethanol for recrystallization. The compound JMlO was obtained as orange crystals (270 mg); mp.137-138 ℃; ESI MS m/z: 425.2[ M + H]⁺；¹H NMR(300MHz，CDCl₃) δ: 10.37(s, 1H, C4-COH), 7.94, 7.71 (two d, each 2H, J ═ 15.6Hz, H-I, 2, 6, 7), 7.26(dd, 2H, J ═ 1.8, 8.7Hz, aromatic 5 ' -H),), 7.17(d, 2H, J ═ 1.8Hz, aromatic 2 ' -H), 6.91(d, 2H, J ═ 8.7Hz, aromatic 6 ' -H), 3.97, 3.95 (two s, each 6H, OCH)₃).

Scheme 7

Synthesis of Compounds Q1OO, Q1O1, JM1, JM6 and JM7

Compounds Q1OO, Q1O1, JM1, JM6 and JM7 were made with an unsaturated side chain at position C4 of ASC-J9 for the purpose of enhancing activity against prostate cancer. Compound Q1OO was prepared by reacting ASC-J9 with 3-bromo-propene in K₂CO₃In the presence of CH at 60 DEG C₂Cl₂The reaction was carried out overnight. Compound Q1O1 was prepared by reacting ASC-J9 with 3-bromo-propene in K₂CO₃And KI at 100 ℃ in DMF for 2 hours (scheme 8). The crude compound was purified by flash column chromatography on silica gel eluting with a mixture of hexane and EtOAc to give the desired product.

Compound Q1OO, yellow amorphous solid, mp.75-78 ℃. ESI MS m/z: 453.1[ M + H]⁺.¹H NMR(300MHz，CDCl₃) δ: 7.75(d, IH, J ═ 15.3Hz, H-I), 7.69(d, IH, J ═ 15.6Hz, H-7), 7.23 to 7.05, and 6.91 to 6.85(m, 7H, aromatic ring H and H-2), 6.73(IH, J ═ 15.6Hz, H-6), 3.96 to 3.91(12H, OCH, H-I)₃) 3.46(IH, C4-H), 2.96(s, IH, acetylene), 2.94-2.90(dd, 2H, -CH)₂CCH)。

Compound Q1O1, amorphous, mp.69-72 ℃ ESI MS m/z: 437.1[ M + H]⁺.¹HNMR(300MHz，CDCl₃) δ: 7.71(d, IH, J ═ 15.6Hz, H-I), 7.70(d, IH, J ═ 15.6Hz, H-7), 7.18 to 7.12(m, 2H, aromatic ring H), 7.06 to 7.00(m, 2H, aromatic ring H), 6.90 to 6.85(2H, aromatic ring H), 6.85(IH, J ═ 15.6Hz, H-2), 6.67(IH, J ═ 15.6Hz, H-6), 5.64 to 5.49(m, IH, ethylene H), 5.19 to 5.07(m, 2H, ethylene H), 3.94 to 3.91(m, 12H, OCH)₃)，296(d，2H，-CH₂-).

Scheme 8

Compound JM1 TotonSynthesized by reacting ASC-J9(40mg) with cinnamyl bromide and anhydrous sodium carbonate in anhydrous acetone (scheme 9). The reaction mixture was heated to reflux for 24 hours. After cooling, the mixture was filtered to remove inorganic solids and the filtrate was evaporated. The crude product obtained was purified by preparative silica gel chromatography plate (n-hexane: ethyl acetate ═ 1: 1) to obtain the desired product JM1 as a light yellow solid. An amorphous state; ESI MS m/z: 513.4[ M + H]⁺；¹H NMR (300MHz, CDCl3) delta: 7.74(d, 2H, J ═ 15.3Hz, H-I, 7), 7.32(d, IH, J ═ 18.6Hz, -CH2CH ═ CH-), 7.4-6.4(1IH, aromatic ring H), 6.93(d, 2H, J ═ 15.3Hz, H-2, 6), 6.46(d, IH, J ═ 18.6Hz, -CH2CH ═ CH-), 3.91, 3.88 (all s, two 6H, OCH ═ CH-), 3.91 (all s, two 6H, OCH, J ═ 18.6Hz, and c, 2, CH ═ CH-)₃)，3.50(br d，2H，-CH2CH＝CH-)。

Compound JM6 was synthesized by reacting ASC-J9(60mg) with bromomethyl acetate (50mg) and sodium hydroxide (20mg) in anhydrous acetone (scheme 9). The reaction mixture was heated to reflux for 24 hours. After cooling, the mixture was filtered to remove inorganic solids and the filtrate was evaporated. The crude product obtained was purified by preparative silica gel chromatography plate (n-hexane: ethyl acetate ═ 1: 2) to obtain the desired product JM6(ESI MSm/z: 467.3[ M + H: 3) as a pale yellow solid]⁺)。

Scheme 9

Compound JM7, obtained as a by-product of JM4 described above, as yellow microcrystals from EtOAc/hexanes; mp.109-110 ℃; ESI MS m/z: 545.2[ M + H]⁺；¹H NMR(300MHz，CDCl₃) δ: 7.83(s, IH, CH ═ C-at C4), 7.79, 7.51, 6.98, 6.83 (all d, IH each, J ═ 15.5Hz, H-I, 2, 6, 7), 7.20, 7.15, 7.08 (all d, IH each, J ═ 1.8, 8.4Hz, aromatic 5 '-H), 6.87(d, IH, J ═ 8.4Hz, aromatic 6' -H), 6.83(d, 2H, J ═ 8.4Hz, aromatic 6 '-H), 7.07, 7.06, 6.99 (all d, IH each, J ═ 1.8Hz, aromatic 2' -H), 3.92, 3.88 (all d, IH, J ═ 1.8Hz, aromatic 2 '-H), 3.88 (all d, IH, each, J ═ 1.8Hz, aromatic 2' -H)Moiety s, 6H, OCH each₃) 3.90, 3.83 (all s, 3H each OCH₃)。

Synthesis of Compounds Q102-Q104, Q108, Q114-Q115, JM12-JM14 and JM16-JM19

The synthesis of compounds Q102-Q104, Q108, JM12-JM14, JM17 was used to evaluate the performance of the C4 substituent on ASC-J9 with different chain lengths, ring sizes and functionalities at the chain ends (e.g. Q108 and JM 14). The synthesis of compounds Q114-Q115, JM16, JM8-19 was used not only to evaluate the C4-side chain, but also to evaluate the effect of substituents on the diphenyl moiety. All compounds are prepared by reacting 2, 4-pentanedione with the appropriate alkyl or alkylene (or substituted alkyl or alkylene) bromide or iodine in benzene, DBU being used as the base. The obtained product 3-substituted 2, 4-pentanedione is further reacted with 3, 4-dimethoxybenzaldehyde or 4-methoxybenzaldehyde or 3-methoxy-4-hydroxybenzaldehyde to obtain the desired product (scheme 10).

An example of the preparation of Q104 is as follows. 0.2g (2mmol) of 2, 4-pentanedione and 30ul (1eq.) of DBU were mixed in 3mL of benzene. To this solution was added dropwise at room temperature 0.48g (1eq.) of a 1ml solution of octyliodobenzene. The resulting solution was stirred at room temperature overnight. The reaction mixture was washed with brine and CH₂Cl₂Extracting with Na₂SO₄Dried and purified by silica gel flash column chromatography to obtain C3-octyl substituted 2, 4-pentanedione and 0-octyl substituted 2, 4-pentanedione. This mixture was reacted with 3, 4-methoxybenzaldehyde by the method described above to obtain compound Q104.

Compound Q104, yellow solid from EtOAc/hexanes (2: 1), mp.87-90 ℃ ESI MSm/z: 509.3[ M + H]⁺；¹H NMR (300MHz, CDCl3) delta: 7.71(d, IH, J-15.6Hz, H-1), 7.63(d, IH, J ═ 15.9Hz, H-7), 7.21-7.14(m, 2H, aromatic H), 7.08-7.05(m, 2H, aromatic H), 6.95(d, IH, J ═ 15.6Hz, H-2), 6.91-6.84(m, 2H, aromatic H), 6.73(d, IH, J ═ 15.9Hz, H-6), 3.94-3.91(m, 12H, -OCH)₃) 2.55(t, IH, H-4), 1.61-1.22(m, 12H, butyl), 0.87(m, 3H, -CH)₃).

Scheme 10

Compound Q102, red needle crystals from EtOAc/hexanes, mp.162-164 ℃ ESI MSm/z: 425.2[ M + H]⁺；¹H NMR (300MHz, CDCl3) delta: 7.73(d, 2H, J ═ 15.3Hz, H-1, 7), 7.23-7.19(dd, 2H, J ═ 8.1, 1.8Hz, aromatic H), 7.09(d, 2H, J ═ 1.5Hz, aromatic H), 6.96(d, 2H, J ═ 15.3Hz, H-2, 6), 6.90(d, 2H, J ═ 8.1Hz, aromatic ring H), 3.96(s, 6H, OCH)₃)，3.94(s，6H，OCH₃)，2.66-2.57(m，2H，-CH₁CH₃)，1.24(t，2H，J＝15.0，6.0Hz，-CH₂CH₃)。℃

Compound Q103, yellow crystals from EtOAc, mp.125-126 ℃ ESI MS m/z: 453.2[ M + H]⁺；¹H NMR (300MHz, CDCl3) delta: 7.74-7.61(2H, H-1, 7), 7.21-7.06(m, 4H, aromatic H), 6.99-6.71(4H, H-2, 6 and aromatic H), 3.94-3.92(12H, -OCH)₃)，2.57(t，IH，H-4)，1.51-1.22(m，6H，-CH₂CH₂CH₂-)，0.87(3H，-CH₃)。

Compound Q108, yellow crystals from EtOAc, mp.60-62 ℃ ESI MS m/z: 515.2[ M + H]⁺；¹H NMR (300MHz, CDCl3) delta: 7.72-7.60(2H, H-1, 7), 7.34-7.00(m, 8H, aromatic H), 6.91-6.84(3H, aromatic H), 6.82-6.68(2H, H-2, 6), 3.95-3.92(12H, -OCH)₃) 3.46(t, IH, H-4), 2.80-2.52(m, 2H, phenyl CH)₂)，2.12-1.84(2H，-CH₂-)，1.68-1.50(2H₁-CHCH₂-)。

Compound JM12, orange town from EtOAc/hexanes, mp.138-139 ℃; ESI MSm/z: 451.2[ M + H]⁺；¹H NMR(300MHz，CDCl₃) δ: 7.72, 6.99 (two d, 2H each, J15.3 Hz, H-I, 2, 6, 7), 7.21(dd, 2H, J1.8, 8.4Hz, aromatic 5 ' -H),), 7.08(d, 2H, J1.8 Hz, aromatic 2 ' -H), 6.90(d, 2H, J8.4 Hz, aromatic 6 ' -H), 5.30(br.s, IH, OH), 3.95, 3.93 (two s, 6H each, OCH, OH)₃)，2.65(d，2H，J＝6.0Hz，C4-CH₂-), 0.95(m, IH, CH for cyclopropane), 0.51, 0.24 (two m, 2H each, CH for cyclopropane)₂)。

Compound JM13, orange needles from EtOAc/hexanes; mp.172-174 ℃; ESI MS m/z: 493.2[ M + H]⁺；¹H NMR(300MHz，CDCl₃) δ: 7.71, 6.97 (two d, 2H each, J15.3 Hz, H-I, 2, 6, 7), 7.20(dd, 2H, J1.8, 8.4Hz, aromatic 5 ' -H),), 7.08(d, 2H, J1.8 Hz, aromatic 2 ' -H), 6.92(d, 2H, J8.4 Hz, aromatic 6 ' -H), 5.30(br.s, IH, OH), 3.95, 3.94 (two s, 6H each, OCH, OH), 3.95 (c, H, OCH, H₃)，2.46(d，2H，J＝6.9Hz，C4-CH₂-), 1.90-1.00(m, 1IH, 1CH and 5CH of cyclopropane)₂).

Compound JM14, orange needles from EtOAc/hexanes; mp.131-132' C; ESI MSm/z: 493.2[ M + H]⁺；¹H NMR(300MHz，CDCl₃) δ: 9.87(br.s, IH, OH), 7.76, 6.90 (two d, 2H each, J15.3 Hz, H-I, 2, 6, 7), 7.19(dd, 2H, J1.8, 8.4Hz, aromatic 5 ' -H),), 7.10(d, 2H, J1.8 Hz, aromatic 2 ' -H), 6.91(d, 2H, J8.4 Hz, aromatic 6 ' -H), 5.30(br.s, IH, OH), 3.95, 3.94 (two s, 6H each, OCH)₃) 2.86, 2.37 (two m, 2H each, C4-CH)₂-CH₂-)。

Compound JM16, orange amorphous; ESI MS m/z: 437.2[ M + H]⁺；¹H NMR(300MHz，CDCl₃Interconversion was observed at 2: 1Isomerism, data listing the main morphology) δ: 7.62, 6.71 (two d, 2H each, J15.9 Hz, H-I, 2, 6, 7), 7.12(dd, 2H, J1.8, 8.4Hz, aromatic 5 ' -H),), 7.05(d, 2H, J1.8 Hz, aromatic 2 ' -H), 6.92(d, 2H, J8.4 Hz, aromatic 6 ' -H), 5.96(br.s, 2H, OH X2), 3.97(s, 3H, OCH X2), 3.97(s, OCH, c, H₃)，3.94(s，9H，OCH₃ X 3)，2.68(d，2H，J＝6.9Hz，C4-CH₂-), 2.19-1.59(m, 7H, 1CH and 3CH for cyclobutane₂)。

Compound JM17, orange needle from EtOAc/hexanes, mp.126-127 ℃; ESI MSm/z: 465.2[ M + H]⁺；¹H NMR(300MHz，CDCl₃And,) δ: 7.72, 7.00 (two d, 2H each, J15.3 Hz, H-I, 2, 6, 7), 7.21(dd, 2H, J1.8, 8.4Hz, aromatic 5 ' -H),), 7.09(d, 2H, J1.8 Hz, aromatic 2 ' -H), 6.92(d, 2H, J8.4 Hz, aromatic 6 ' -H), 3.96, 3.95 (two s, 6H each, OCH₃)，2.70(d，2H，J＝6.9Hz，C4-CH₂-, 2.08(m, 2H, 1CH for cyclobutane)₂) 1.83(m, 4H, 2CH for cyclobutane)₂)。

Compound JM18, orange amorphous; ESI MS m/z: 423.2[ M + H]⁺；¹H NMR(300MHz，CDCl₃Tautomerism was observed at 2: 1, data for the major forms are presented) δ: 7.65, 6.73 (two d, 2H each, J15.9 Hz, H-I, 2, 6, 7), 7.12(dd, 2H, J1.8, 8.4Hz, aromatic 5 ' -H),), 7.05(d, 2H, J1.8 Hz, aromatic 2 ' -H), 6.92(d, 2H, J8.4 Hz, aromatic 6 ' -H), 3.93(s, 12H, OCH)₃X4), 2.71, 2.65 (two d, each 1H, J ═ 6.0Hz, C4-CH₂-), 0.95(m, IH, CH for cyclopropane), 0.51, 0.24 (two m, 2H each, CH for cyclopropane)₂)。

Compound JM19, orange-red needles from EtOAc/hexanes; mp.153-154 deg.C; ESI MSm/z: 465.2[ M + H]⁺；¹H NMR(300MHz，CDCl₃And,) δ: 7.75, 6.88 (two d, 2H each, J15.5 Hz, H-I, 2, 6, 7), 7.17(dd, 2H, J1.6, 8.5Hz, aromatic 5' -H),), 7.07(d, 2H, J ═ 1.6Hz, aromatic 2 '-H), 6.96(d, 2H, J ═ 8.5Hz, aromatic 6' -H), 5.90(s, 2H, OHX 2), 3.96(s, 6H, OCH)₃X T), 2.86, 2.35 (two m, 2H each, C4-CH)₂-CH₂-)。

Compound Q114, yellow crystalline solid from EtOAc/hexanes, mp.166-167

Of C.ESI MS m/z：337.0[M+H]ι+；.1¹H NMR (300MHz, CDCl3) delta: 7.63(d, 2H, J ═ 16.1Hz, H-1, 7), 7.53-7.50(m, 4H, aromatic H), 6.94-6.91(m, 4H, aromatic H), 6.50(d, 2H, J ═ 16.1Hz, H-2, 6), 5.79(s, IH, H-4), 3.85(s, 6H, OCH)₃)。

Compound Q115, yellow crystals from EtOAc mp.142-143 ℃ ESI MS m/z:

393.1[M+H]⁺；¹h NMR (300MHz, CDCl3) delta: 7.73(d, 2H, J ═ 15.6Hz, H-1, 7), 7.55-7.52(4H, aromatic H), 6.99-6.92(6H, H-2, 6 and aromatic H), 3.86(6H, -OCH)₃)，2.55(t，IH，H-4)，1.53-1.40(m，6H，-CH₂CH₂CH₂-)，1.01(t，3H，-CH₃)。

Synthesis of the Compounds: JM4, JM20, and Q116

The compounds JM4, JM20 and Q116 have the common performance of (substituted) -triaryl system [ tri (substituted phenyl) acrolein conjugated bond ]. One of the purposes of synthesizing these compounds was to investigate the effect of the polyphenylacrolein moiety on anti-AR and anti-prostate cancer activity. Compound JM4 was synthesized by polycondensation of 3, 4-dimethoxybenzaldehyde and triacetylmethane as shown in scheme 11.

Compounds JM20 and Q116 were synthesized by the same method as described for JM 4.

Compound JM20, red powder, mp.165-167 ℃; ESI MS m/z: 455.2[ M + H]⁺；¹H NMR(300MHz，d₆-DMSO,) δ: 7.69(d, 2H, J-15.6Hz, H-1, 7), 7.03(d, IH, J-16.2 Hz, side C4)chain-COCH ═ CH-), 7.62-7.34(m, 6H, aromatic ring H), 6.67(d, IH, J ═ 15.6Hz, C4 side chain-COCH ═ CH-), 6.90-6.72(m, 4H, aromatic H), 6.57(d, 2H, J ═ 15.9Hz, H-2, 6).

Compound Q116, yellow amorphous solid, mp.70-72 ℃ ESI MS m/z: 497.1[ M + H]⁺；¹H NMR (300MHz, CDCl3) delta: 7.78(d, 2H, J ═ 15.3Hz, H-1, 7), 7.60(d, IH, J ═ 15.6Hz, C4 side chain-COCH ═ CH-), 7.54-7.51(2H, aromatic ring H), 7.47-7.44(4H, aromatic ring H), 6.97(d, IH, J ═ 15.6Hz, C4 side chain-COCH ═ CH-), 6.92-6.85(6H, aromatic H), 6.71(d, 2H, J ═ 15.3Hz, H-2, 6), 3.84-3.82(9H, -OCH) (— OCH 9H ═ OCH)₃)。

Scheme 11

Compound JM4

Synthesis of the Compounds: JM5

Compound JM5, structurally containing four (substituted phenyl) acrolein moieties, was synthesized by reacting ASC-J9(18.9g) with bromomethyl acetate (10.0g) in the presence of sodium carbonate (5.0g) in anhydrous acetone (250mL) (scheme 12). After heating at reflux for 80h, the solid was filtered and the filtrate was concentrated in vacuo. The residue was subjected to repeated silica gel column chromatography (n-hexane: ethyl acetate 2: 1) to obtain the desired product and recover the starting material ASC-J9(15 g). The product obtained is dissolved in 0.5ml of ethyl acetate and added dropwise to 5ml of hexane with stirring. Filtration and vacuum drying gave JM5(877mg) as a yellow powder.

Compound JM5 can also be synthesized by a short time, high yield reaction of ASC-J9 with bromomethyl ether and sodium carbonate in anhydrous acetone.

Scheme 12

Compound JM5

Compound (I)JM5The analytical data of (2) are shown below.

Yellow amorphous, mp.111-114 ℃ ESI MS m/z: 804.87[ M + H]⁺(ii) a On 500MHzVarian¹H and¹³the C NMR data (CDCl3) are shown in Table 1.

TABLE 1 ASC-JM5¹H and¹³c NMR spectrum data.

No.	Groups	δ1H	δ13C
				a1	Ar.＝C		126.83
a2	Ar.＝CH	7.098(d，J＝1.5)	109.62
				a3，4，b3，4，c3，c4，d3，4.	Ar.＝C(OCH3)	*3.812(s)，3.596，3.708，3.968，3.950，3.946，3.857，3.931	147.77，147.93，148.96，149.15，149.17，151.25，151.69，151.87
a5	Ar.＝CH	6.878(d，J＝8.0)	110.82
				a6	Ar.＝CH	7.167(dd，J＝8.0，1.5)	123.31
a11	＝CH	7.732(d，J＝15.5)	143.25
				a12	＝CH	7.017(d，J＝15.5)	116.46
a13	C＝O		182.21
				a14	-CH-	4.317(s)	40.75
b1	Ar.＝C		127.84
				b2	Ar.＝CH	6.881(d，J＝2.0)	109.57
b5	Ar.＝CH	6.740(d，J＝8.0)	110.71
				b6	Ar.＝CH	7.061(dd，J＝8.0，2.0)	124.09
b11	＝CH	7.681(d，J＝15.5)	145.83
				b12	＝CH	6.739(d，J＝15.5)	118.33
b13	C＝O		196.29
				c1	Ar.＝C		126.80
c2	Ar.＝CH	7.055(d，J＝1.5)	109.80
				c5	Ar.＝CH	6.878(d，J＝8.5)	110.82
c6	Ar.＝CH	7.195(dd，J＝8.5，1.5)	123.84
				c11	＝CH	7.60(d，J＝15.5)	144.71
c12	＝CH	7.038(d，J＝15.5)	119.03
				c13	C＝O		194.59
c14	-CH-	4.317(d，J＝7.5)	40.69
				d1	Ar.＝C		133.03
d2	Ar.＝CH	6.653(s)	111.92
				d5	Ar.＝CH	6.740(d，J＝8.5)	110.71
d6	Ar.＝CH	6.676(d，J＝8.0)	120.28
				d11	-C-		68.86
d12	-CH2	3.268(d，J＝16.5)，3.352(d，J＝16.5)	24.13
				d13	C＝O		190.32
d14	-CH2	2.734(d，J＝19.5)，3.516(dd，J＝19.5，7.5)	37.74
				a3，4，b3，4，c3，c4，d3，4.	-OCH 3	3.596(s)，3.708(s)，3.812(s)，3.857(s)，3.946(s)，3.931(s)，3.950(s)，3.968(s)	55.4055.5955.8755.9456.05

^*1H data of methoxy proton (Note: Group-Group)

Synthesis of the Compounds: q110 and Q111

In order to investigate the effect of the length of the conjugated bridge on the AR activity, a compound Q110 having four-conjugated double bond linkages and a compound having five-conjugated double bond linkages were synthesizedThe terminal compound Q111 is also illustrated in scheme 13. Compound Q110 was synthesized starting from 1, 2-dimethoxy-4-propylbenzene. To a solution of 3- (3, 4-dimethoxyphenyl) propane in anhydrous dioxane was added DDQ (3.1eq.) and a catalytic amount of acetic acid. The mixture was sonicated for 2h under TLC control. After completion of the reaction, the solid was filtered off and the filtrate was concentrated. The residue was dissolved in EtOAc and washed with water, 2% NaHCO₃And a brine rinse. Subjecting the organic extract to Na₂SO₄Drying and concentration gave a yellow-brown solid crude which was purified by neutral alumina column chromatography and eluted with hexane-ethyl acetate mixture to give a pale yellow solid, 3, 4-dimethoxycinnamaldehyde, in 60% yield (b.p. joshi et al, Tetrahedron, 62, 2590-. 2, 4-pentanedione (3eq.) and B₂O₃(1eq.) of EtOAc solution was stirred at 40 ℃ for 0.5h and 3, 4-dimethoxycinnamaldehyde (1eq.) and tributylborane (1eq.) were added. The reaction mixture obtained was stirred at 40 ℃ for 0.5 h. At this temperature a solution of butylamine (1.2eq.) in EtOAc was added dropwise and stirred at 4 ℃ for 16 h. To the red reaction mixture, 1% aqueous HCl solution was added and the mixture was stirred for 1 hour to 60 ℃. After cooling to room temperature, the aqueous phase was separated and the organic phase was washed with water to pH-7 and dried over Na2SO 4. The crude product was purified by silica gel flash column chromatography to afford the intermediate 8- (3, 4-dimethoxy-phenyl) -4 hydroxy-octa-3, 5, 7-triethylenetetramine-2-one as an off-white solid. The intermediate (1eq.) and B were stirred at 70 ℃₂O₃(0, 7eq.) in EtOAc for 0.5 h. 3, 4-dimethoxybenzaldehyde (1eq.) and tributylborane (1eq.) were added and the reaction mixture was stirred at 70 ℃ for 0.5 h. Piperidine (1.2eq.) in EtOAc solution was added dropwise and the reaction mixture was stirred at 88-90 ℃ for 1 h. After cooling to 60 ℃ 1% aqueous HCl was added and the mixture was stirred at 60 ℃ for 0.5 h. The reaction mixture was obtained stepwise by following the procedure described above and the crude product was purified by silica gel column chromatography to obtain the desired product Q110 as a red solid. Amorphous state, mp.65-68 ℃, ESI MS m/z: 423.1[ M + H]⁺；¹H NMR (300MHz, CDCl3) delta: 7.64-7.58(d, 2H, H-I and 2), 7.16-7.02(4H, aromatic ring H and retro double bond H), 6.90-6.82(4H, aromatic ring H), 6.53-6.48(IH, retro double bond H), 6.18-6.12(IH, retro bisBond H), 5.75(s, IH, H-4), 3.94-3.92(12H, -OCH)₃)。

Compound Q111 was synthesized by reacting 8- (3, 4-dimethoxy-phenyl) -4-hydroxy-octa-3, 5, 7-triethylenetetramine-2-one (3) with 3, 4-dimethoxycinnamaldehyde (scheme 13) as described in the synthesis of Q110. A red amorphous solid was obtained, mp.187-9 ℃ ESI MS m/z: 449.1[ M + H]⁺；¹HNMR (300MHz, CDCl3) δ: 7.49-7.40(d, 2H, H-I and 11), 7.06-7.02(4H, aromatic ring H), 6.87-6.81(2H, aromatic ring H, and 4H for the retro double bond H), 6.17-6.12(2H, retro double bond H), 5.75(s, IH, H-4), 3.94-3.92(12H, -OCH)₃)。

Scheme 13

Example 2: biological effects of compounds having at least one (3, 4-alkoxy or hydroxy-substituted phenyl) acrolein moiety on human Androgen Receptor (AR) and androgen/AR-modulating activity were examined.

Their activity in inhibiting androgen/AR-induced function was tested against typical ASC compounds and monomers. Cell growth assays were performed using human prostate cancer cells, either LNCaP or CWR22 Rvl. Functional AR proteins are expressed in two cell lines, however the growth of LNCaP cells is DHT dependent, but not of CWR22Rvl cells derived from refractory worsening hormone tumors. Furthermore, western blot analysis was performed by testing monomers, and some typical novel compounds in prostate cancer cells, and the results showed that compounds having at least one (4-hydroxy-3-methoxy-phenyl) acrolein moiety were able to reduce AR protein expression levels and inhibit cancer cell growth in vitro.

In vitro cell growth assays using human prostate cancer cell lines, LNCaP and CWR22Rvl

The present invention performs a proliferation assay of MTT cells to determine the ability of a compound to suppress or inhibit the growth of prostate cancer cells. The MTT assay, a widely used method to measure proliferation of cultured cells and relies on the conversion of colourless medium to tetrazolium reduced by mitochondrial dehydrogenases (all viable cells have), and has been previously shown (Su et ai, 1999) to evaluate the growth of various tissue culture cells. Briefly, 1X 10 suspended in the entire medium³Was dispensed into each well of a 96-well micro-test III tissue culture dish (Falcon, NJ). After two days, the medium was replaced with RPMI-1640 medium containing 10% charcoal/FBS derived from dextran (hormone-derived fetal bovine serum). The compounds tested were added to the medium at the indicated concentrations, with or without 1nM DHT, and the cells were incubated in an incubator (37 ℃) for 5 days. 2 hours prior to harvest, 1/10 volumes of MTT medium solution (5mg/ml PBS) were added to the cells in each box. After 2 hours incubation, the dishes were centrifuged (10 minutes, 1,000rpm) and the supernatant carefully removed from each well. Mu.l of lysis buffer (50% dimethylformamide, 5% sodium dodecyl sulfate, 0.35M acetic acid, and 50mM HCl) was added to each well to lyse the cells and the lysed tetrazole in each well. The relative enzyme activity of each grid was measured from absorbance readings at 450nm wavelength using a Bio-RAD reference microplate reader. Data from the MTT assay were also confirmed by actual cell number and cell morphology on parallel standing separation dishes. Data from the parallel dishes indicate a direct relationship between the number of enzyme activities and the number of viable cells in the grid.

Western blot analysis of AR protein expression levels in prostate cancer cells

Widely used western blot analysis was used to measure AR protein expression levels. Human prostate cancer cells, LNCaP and cwr22rvl, both expressed high amounts of AR protein and were used in this study. In the present invention, typical ASC compounds were tested in western blot assays to evaluate their activity in reducing AR expression; and the assay was performed in the presence or absence of dihydrotestosterone (DHT, 1 nM). After incubating the cells with the test compound for the indicated time, they are collected and lysed according to western blot techniques known in the biochemical art. Details of western blot analysis methods have been previously published (Su et al, 1999). Briefly, cells were collected in 2X sodium dodecyl sulfate/polyacrylamide gel electrophoresis loading buffer or in radioimmunoprecipitation assay (RIPA) buffer fortified with 10. mu.g/ml benzamidine, 10. mu.g/ml trypsin inhibitor, and 1mM phenylmethylsulfonyl fluoride. A sample of total protein from each cell lysate (about 40. mu.g) was separated by electrophoresis on an SDS/PAGE gel. Following electrophoretic separation, proteins were transferred from the gel to nitrocellulose membranes following standard procedures. The membranes were then incubated with 10% non-fat milk in phosphate buffered saline supplemented with 0.1% Tween20(PBST) for 1 hour, followed by overnight incubation with primary human AR-specific antibody (purchased from BD-PharMingen) at 4 ℃. After incubation, membranes were washed 3 times with PBST buffer for 10 min each; alkaline phosphatase conjugated secondary antibody was then added and incubated at room temperature for 1 hour. After the second incubation, the membrane was washed again with PBST and the AR-protein signal in the membrane was visualized by adding alkaline phosphatase medium, bromochloroindolyl phosphate and nitrotetrazole to the membrane. To ensure that equal amounts of protein were analyzed in each sample, a portion of the membrane was kept with specific antibodies for the management of the protein β -actin (santa cruz biotechnology) and actin signals were displayed with the second antibody described above. Protein signal intensity (as indicated by the color band on the membrane) was measured using a densitometer and analyzed by NIH image J software (NIH 1.33). The amount of AR-protein was calculated by normalizing the amount of AR to the amount of β -actin in each sample and expressed as relative values.

Detection of AR degradation using cycloheximide tracking test method

The "degradation" of AR protein in prostate cancer cells was measured by using a cycloheximide (protein synthesis inhibitor) follow-up test method. Briefly, LNCaP cells were incubated with test ASC compounds for 24 hours at the indicated concentrations. Subsequently, cycloheximide was added to the cells at a concentration of 15. mu.g/ml to inhibit new protein synthesis. After incubation, cells were harvested at the indicated time periods and analyzed for changes in the results of AR protein levels using western blot analysis as described above.

Claims (12)

1. A compound selected from the group consisting of II-1, II-2 and II-3:

2. a compound according to formula IIc:

wherein:

R₃、R₄、R₃' and R₄' are all independently selected from the group consisting of-H, -OH, and-OCH₃In the group consisting of; and is

R₁And R₂Independently selected from the group consisting of-H, -CH₃and-C₂H₅In the group consisting of.

3. A compound according to formula III:

wherein R is₃、R₄、R₃’、R₄’、R₃"and R₄"is independently selected from the group consisting of alkoxy, hydroxy, and hydrogen.

4. A compound according to formula V:

wherein:

each "n" is independently 2 or 3;

R₃,R₄,R₃' and R₄' is independently selected from the group consisting of-H, -OH and-OCH₃In the group consisting of.

5. A pharmaceutical composition comprising a compound of claim 1 in a pharmaceutical carrier.

6. A pharmaceutical composition comprising a compound of claim 2 in a pharmaceutically acceptable carrier.

7. A pharmaceutical composition comprising a compound of claim 3 in a pharmaceutically acceptable carrier.

8. A pharmaceutical composition comprising the compound of claim 4 in a pharmaceutical composition.

9. Use of a compound of any one of claims 1, 2, 3, or 4 in the manufacture of a medicament for treating a subject suffering from an androgen associated medical condition, comprising administering to a subject suffering from an androgen associated medical condition a compound of any one of claims 1, 2, 3, or 4 optionally with a pharmaceutically acceptable carrier.

10. Use of a compound of any one of claims 1, 2, 3, or 4 in the manufacture of a medicament for treating a subject suffering from kennedy's disease, comprising administering to a subject suffering from kennedy's disease a compound of any one of claims 1, 2, 3, or 4 optionally with a pharmaceutically acceptable carrier.

11. Use of a compound of any one of claims 1, 2, 3, or 4 in the manufacture of a medicament for treating a patient having cancer, comprising:

a) providing a patient having a cancer selected from the group consisting of prostate cancer, bladder cancer, liver cancer, and breast cancer;

b) administering to said patient a therapeutically effective amount of a compound of any one of claims 1, 2, 3 or 4, optionally with a pharmaceutically acceptable carrier.

12. The use according to claim 9, wherein the androgen associated medical condition is selected from the group consisting of androgen associated inflammation, acne, alopecia, hirsutism and wound.