US20030144346A1 - Methods and compositions for regulation of 5-alpha reductase activity - Google Patents

Methods and compositions for regulation of 5-alpha reductase activity Download PDF

Info

Publication number
US20030144346A1
US20030144346A1 US10/294,331 US29433102A US2003144346A1 US 20030144346 A1 US20030144346 A1 US 20030144346A1 US 29433102 A US29433102 A US 29433102A US 2003144346 A1 US2003144346 A1 US 2003144346A1
Authority
US
United States
Prior art keywords
subject
compound
compounds
reductase
activity
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Abandoned
Application number
US10/294,331
Inventor
Shutsung Liao
Richard Hiipakka
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
CHICAGO OFFICE OF TECHNOLOGY AND INTELLECTUAL PROPERTY THE, University of
Original Assignee
CHICAGO OFFICE OF TECHNOLOGY AND INTELLECTUAL PROPERTY THE, University of
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by CHICAGO OFFICE OF TECHNOLOGY AND INTELLECTUAL PROPERTY THE, University of filed Critical CHICAGO OFFICE OF TECHNOLOGY AND INTELLECTUAL PROPERTY THE, University of
Priority to US10/294,331 priority Critical patent/US20030144346A1/en
Assigned to UNIVERSITY OF CHICAGO OFFICE OF TECHNOLOGY AND INTELLECTUAL PROPERTY, THE reassignment UNIVERSITY OF CHICAGO OFFICE OF TECHNOLOGY AND INTELLECTUAL PROPERTY, THE ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: HIIPAKKA, ROBERT, LIAO, SHUTSUNG
Publication of US20030144346A1 publication Critical patent/US20030144346A1/en
Abandoned legal-status Critical Current

Links

Images

Classifications

    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00Medicinal preparations containing organic active ingredients
    • A61K31/33Heterocyclic compounds
    • A61K31/395Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44Non condensed pyridines; Hydrogenated derivatives thereof
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00Medicinal preparations containing organic active ingredients
    • A61K31/33Heterocyclic compounds
    • A61K31/395Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/55Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having seven-membered rings, e.g. azelastine, pentylenetetrazole
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00Medicinal preparations containing organic active ingredients
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00Medicinal preparations containing organic active ingredients
    • A61K31/075Ethers or acetals
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00Medicinal preparations containing organic active ingredients
    • A61K31/12Ketones
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00Medicinal preparations containing organic active ingredients
    • A61K31/33Heterocyclic compounds
    • A61K31/335Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/35Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P13/00Drugs for disorders of the urinary system
    • A61P13/08Drugs for disorders of the urinary system of the prostate
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00Drugs for dermatological disorders
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00Drugs for dermatological disorders
    • A61P17/14Drugs for dermatological disorders for baldness or alopecia
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00Drugs for disorders of the metabolism
    • A61P3/04Anorexiants; Antiobesity agents
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00Antineoplastic agents
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07CACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C49/00Ketones; Ketenes; Dimeric ketenes; Ketonic chelates
    • C07C49/20Unsaturated compounds containing keto groups bound to acyclic carbon atoms
    • C07C49/24Unsaturated compounds containing keto groups bound to acyclic carbon atoms containing hydroxy groups
    • C07C49/245Unsaturated compounds containing keto groups bound to acyclic carbon atoms containing hydroxy groups containing six-membered aromatic rings
    • C07C49/248Unsaturated compounds containing keto groups bound to acyclic carbon atoms containing hydroxy groups containing six-membered aromatic rings having unsaturation outside the aromatic rings
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07CACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C49/00Ketones; Ketenes; Dimeric ketenes; Ketonic chelates
    • C07C49/20Unsaturated compounds containing keto groups bound to acyclic carbon atoms
    • C07C49/255Unsaturated compounds containing keto groups bound to acyclic carbon atoms containing ether groups, groups, groups, or groups
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07DHETEROCYCLIC COMPOUNDS
    • C07D311/00Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
    • C07D311/02Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
    • C07D311/04Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
    • C07D311/58Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4
    • C07D311/60Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4 with aryl radicals attached in position 2
    • C07D311/62Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4 with aryl radicals attached in position 2 with oxygen atoms directly attached in position 3, e.g. anthocyanidins

Definitions

  • the present invention relates generally to compounds, compositions and methods regulating the action and function of androgens and other steroid hormones by modulating the activity of steroid-reductases, including isozymes of 5 ⁇ -reductases. More specifically, the present invention relates to the use of these compounds to regulate processes or treat disorders that are modulated by androgens or other steroid hormones or are caused by abnormal actions of androgens or other steroid hormones in cells or organs of animals, humans, plants, or microorganisms.
  • This invention relates to the use of natural and synthetic flavanoids, catechols, curcumin-related substances, quinones, catechins and fatty acids and their analogues or derivatives as 5 ⁇ -reductase isozyme inhibitors and as therapeutic agents. These compounds can also be used in promoting or modulating desirable production of specific products for commercial purposes.
  • testosterone is converted to a more active metabolite 5 ⁇ -dihydrotestosterone (DHT) by 5 ⁇ -reductase (Anderson and Liao, 1968; Bruchovsky and Wilson, 1968).
  • DHT 5 ⁇ -dihydrotestosterone
  • Other substrates of 5 ⁇ -reductases are also converted to reduce products that may have specific properties.
  • Inhibition of 5 ⁇ -reductase represents a unique approach for developing therapeutic methods for androgen-dependent diseases, such as benign prostatic hyperplasia, breast and prostatic cancer, skin disorders, seborrhea, common baldness, hirsutism, and hidradenitis suppurative.
  • Certain aliphatic unsaturated fatty acids such as ⁇ -linolenic acid (Liang and Liao, 1992) and catechin-3-gallates (Liao and Hiipakka, 1995), can inhibit 5 ⁇ -reductase activity of liver and prostate of rats and humans in vitro.
  • 5 ⁇ -Reductase is found in many organs (Russell and Wilson, 1994; Hiipakka et al., 1993) including the sebaceous gland of hamsters (Takayasu and Adachi, 1972) and human hair follicles (Randall, 1994).
  • Two 5 ⁇ -reductase isozymes have been identified in rats and humans (Russell and Wilson, 1994).
  • the type 1 isozyme predominates in rat tissues such as liver, kidney, brain, and lung, whereas the type 2 enzyme is more abundant in rat testis and epididymis. Both isozymes are found in skins of the neonate, but the type 1 isozyme is the major form expressed in the skin after puberty.
  • the type 1 isozyme is also expressed in balding scalp.
  • Finasteride a 4-azasteroid, is a competitive inhibitor of 5 ⁇ -reductases and has an affinity 30-fold higher for isozyme 2 than for isozyme 1 (Russell and Wilson, 1994).
  • the green tea catechins, epicatechin-3-gallate and epigallocatechin-3-gallate are more effective inhibitors of the type 1 enzyme and ⁇ -linolenic acid inhibits both isozymes equally well (Liao and Hiipakka, 1995).
  • Male-patterned baldness can start as early as the teens in genetically susceptible males, and it has been estimated to be present in 30% of Caucasian males at age 30, 40% of Caucasian males at age 40, and 50% of Caucasian males at age 50.
  • Acne is the most common skin disorder treated by physicians.
  • hirsutism is one of the hallmarks of excessive androgen. The ovaries and the adrenal are the major sources of androgen in women.
  • testosterone the major androgen circulating in the blood is testosterone.
  • About 98% of the testosterone in blood is bound to serum proteins (high affinity finding to sex-steroid binding globulin and low affinity binding to albumin), with only 1-2% in free form.
  • the albumin-bound testosterone, the binding of which is readily reversible, and the free form are considered to be bioavailable, and account for about 50% of total testosterone.
  • Testosterone enters target cells apparently by diffusion. In the prostate, seminal vesicles, skin, and some other target organs, it is converted by a NADPH-dependent 5 ⁇ -reductase to a more active metabolite, 5 ⁇ -DHT.
  • 5 ⁇ -DHT then binds an androgen receptor (AR) in target organs.
  • AR androgen receptor
  • the 5 ⁇ -DHT-receptor complexes interact with specific portions of the genome to regulate gene activities (Liao et al., 1989).
  • Testosterone appears to bind to the same AR, but it has a lower affinity than 5 ⁇ -DHT.
  • tissues such as muscle and testes, where 5 ⁇ -reductase activity is low, testosterone may be the more active androgen.
  • 5 ⁇ -DHT Excessive 5 ⁇ -DHT is implicated in certain androgen-dependent pathological conditions including BPH, acne, male-pattern baldness, and female idiopathic hirsutism. It has been shown that 5 ⁇ -reductase activity is reported to be higher in hair follicles from the scalp of balding men than that of non-balding men.
  • the present invention relates generally to the utilization of certain compounds for the control of androgen activity in target organs and cells through the modulation of a 5 ⁇ -reductase activity.
  • these compounds are employed to repress androgenic activity by inhibiting the formation and availability of active androgen in target cells. Consequently, the present invention is useful for the treatment of a wide variety of conditions including, but not limited to, the treatment of prostatic hyperplasia, prostatic cancer, hirsutism, acne, male pattern baldness, seborrhea, and other diseases related to androgen hyperactivity.
  • Compounds useful in the practice of the present invention include various isomers of saturated and unsaturated fatty acids, natural and synthetic analogues, and derivatives from which these fatty acids can be generated as well as the metabolites and oxidation products of these fatty acids. The use of these and other fatty acids and their derivatives is also contemplated. Also useful are catechin compounds, particularly, catechins that are structurally similar to epicatechin gallate (ECG) and epigallocatechin gallate (EGCG). EGCG has an additional hydroxyl group on the epicatechin gallate molecule, which has been found to be surprisingly active in modulating several 5 ⁇ -reductase mediated processes.
  • ECG epicatechin gallate
  • EGCG epigallocatechin gallate
  • EGCG derivatives having such an additional OH group on the altering ECG molecule were shown to be active in inducing body weight loss and particularly in reducing the size of androgen sensitive organs such as preputial glands, ventral prostate, dorsolateral prostate, coagulating glands, seminal vesicles, human prostate tumors, and breast tumors in nude mice.
  • catechins particularly EGCG
  • prostate weight loss was percentage-wise more than three times as much.
  • the loss in body weight and the organ weight are likely due to EGCG interference of a common step in the pathway controlling body weight and the organ weight gain.
  • EGCG and related compounds may interact and interfere with a receptor macromolecule (probably containing a protein) that modulates specific lipid synthesis and accumulation. Lipids can modulate gene expression, cell development and differentiation, and organ growth.
  • Specific interference of lipid metabolism in the cells and organs may control the growth of the organs, in particular, prostate, sebaceous, preputial and other secretory organs. In certain applications, it is expected that benign or abnormal growth or cancer of these organs may be treated or even prevented by administration of catechin related compounds.
  • catechin compounds will arrest or reduce human prostate and breast cancer cell growth.
  • the effectiveness of catechin compounds was shown to be dependent on the methods by which these compounds were administered to the experimental animals. Intraperitoneal application was much more effective than oral administration. It is expected that direct application to the organs, such as the prostate, will be very effective.
  • EGCG was surprisingly effective in suppressing and even reducing the size of human prostate and breast tumors in animal models. The effect was illustrated with EGCG; however, structurally similar catechin compounds are also effective, particularly those that are structurally similar to EGCG in having at least one additional hydroxyl group as compared with ECG.
  • the EGCG species that contain eight hydroxyl groups is significantly more effective in reducing body weight than is ECG, which contains seven hydroxyl groups.
  • Compounds of this general structure are expected to be particularly effective in chemoprevention and chemotherapy of human prostate cancer.
  • Compounds having a part of structure similar to a part of structure of EGCG are also expected to be effective.
  • Compounds can be used as antiandrogenic agents through topical or systemic application.
  • a preparation for this purpose can include a carrier, a protectant, an antioxidant (such as vitamin C or E, and various catechins and polyphenols), and other pharmaceutical and pharmacological agents. It is also expected that such compounds can be used in a delivery system (oral, local application, injection, or implantation) involving molecular recognition through which the compounds are delivered to target sites. Such a delivery system may involve, among other methods, liposome techniques or immunological devices.
  • Natural or synthetic chemicals that can modulate the production or cellular action of receptors and macromolecules are useful in the treatment of abnormalities such as obesity, BPH, prostate cancer, skin diseases, baldness, breast tumors, and hirsutism, which are related to lipid synthesis, body weight, and/or androgen function.
  • Animal models can be used to demonstrate the effectiveness of compounds on a variety of cancers.
  • Shionogi tumor and other tumors can be studied in male rats.
  • Human breast and prostate cancer cell growth can be studied in nude mice.
  • rodent breast tumors induced by carcinogens and other cancers induced in transgenic mice or Dunning tumor in rats can be similarly analyzed for their chemotherapy by EGCG and related compounds.
  • disorders include, but are not necessarily limited to, those conditions wherein excessive androgenic activities have been implicated, for example, male pattern baldness, female hirsutism, skin disorders, BPH, cancers of prostate, breast, skin and other organs.
  • the present invention is also directed to novel compounds. These compounds have the formula:
  • R 1 , R 2 and R 3 each may be —H, —OH or —OCH 3 , provided that only one of R 1 , R 2 , and R 3 may be —H;
  • R 4 , R 5 and R 6 each may be —H, —OH, —OCH 3 or —N(CH 3 ) 2 , provided that only one of R 4 , R 5 and R 6 may be —H;
  • n 0 or 1.
  • compositions and methods disclosed and claimed herein can be made without undue experimentation in light of the present disclosure. While the compositions and methods of this invention have been described in terms of preferred embodiments, it will be apparent to those of skill in the art that variations may be applied to the compositions, methods and in the steps or in the sequence of steps of the method described herein without departing from the concept, spirit and scope of the invention. More specifically, it will be apparent that certain agents which are both chemically and physiologically related may be substituted for the agents described herein while the same or similar results would be achieved. All such similar substitutes and modifications apparent to those skilled in the art are deemed to be within the spirit, scope and concept of the invention as defined by the appended claims.
  • FIG. 1 shows the structure of flavanoid compounds of the present invention.
  • FIG. 2 shows the structure of catechol compounds of the present invention.
  • FIG. 3 shows the structure of curcumin and related compounds of the present invention.
  • FIG. 4. shows quinones of the present invention.
  • FIG. 5 shows epigallocatechin derivative compounds of the present invention.
  • FIG. 6 shows the generic formula of the epigallocatechin derivatives of the present invention
  • FIG. 7 shows the generic formula of gallates useful in the present invention
  • FIG. 8 shows the generic formula of curcumin derivatives useful in the present invention
  • FIG. 9 shows the generic formula of quinones and catechols useful in the present invention.
  • FIG. 10 shows fatty acids of the present invention.
  • the present invention is concerned with methods of inhibiting 5 ⁇ -reductase, which include subjecting a cell to an effective concentration of a 5 ⁇ -reductase inhibitor such as one of the compounds disclosed herein. It is believed that the use of such inhibitors to block abnormal androgen action will serve to treat cancer in conjunction with other anti-cancer agents, chemotherapy, resection, radiation therapy, and the like.
  • the compounds of this invention besides acting as 5 ⁇ -reductase inhibitors, may have other effects that can lead to antitumor activity or to suppress abnormal growth of prostate or other organs.
  • 5 ⁇ -reductase In mammalian cells, 5 ⁇ -reductase is very tightly associated with intracellular membranes, including the membrane of the endoplasmic reticulum and contiguous nuclear membranes. Attempts to solubilize and purify active 5 ⁇ -reductase have not been very successful.
  • the assay of 5 ⁇ -reductase activity therefore, is performed by measuring the rate of conversion of testosterone to 5 ⁇ -DHT by whole cells or by microsomal and nuclear preparations in the presence of NADPH (enzymatic assay).
  • the 5 ⁇ -reductase activity can be reliably assayed by following NADPH-dependent noncovalent binding of a potent radioactive inhibitor, such as [ 3 H]4-MA ([ 3 H]4-MA-binding assay), which strongly competes with testosterone for binding to the reductase.
  • a potent radioactive inhibitor such as [ 3 H]4-MA ([ 3 H]4-MA-binding assay)
  • the results of the two assays correlate very well when microsomal preparations from different organs or animals are used for comparison.
  • the binding assay solution in a final volume of 0.15 ml, contains microsomes (2-20 ⁇ g of protein), 0.07 ⁇ Ci of [ 3 H]4-MA, 0.1 mM-NADPH, 1 mM-dithiothreitol and 50 mM-potassium phosphate, pH 7.0, with or without the indicated amount of a lipid or an inhibitor preparation.
  • Lipids are dissolved in ethanol and added in 1-5 ⁇ l volumes. Control tubes receive the same amount of ethanol. After incubation at 0° C.
  • the [ 3 H]4-MA bound to microsomes is determined by collecting microsomes on a Whatman glass fibre filter and washing with 10 ml of 20 mM-potassium phosphate, pH 7.0, containing 0.01% CHAPS to remove unbound [ 3 H]4-MA.
  • the standard reaction mixture in a final volume of 0.15 ml, contains microsomes, 1 ⁇ Ci of [ 3 H]testosterone, 0.5-3.0 ⁇ M non-radioactive testosterone, 0.1 mM-NADPH, 1 mM-dithiothreitol and 50 mM-potassium phosphate, pH 7.0, with or without the indicated amount of a lipid or an inhibitor preparation.
  • the reaction is started by the addition of microsomes and the incubation is carried out at 37° C for 15 minutes. Steroids are extracted and separated by thin layer chromatography. Radioactive steroids are located by fluorography and the amount of radioactivity present determined by scintillation counting. The 5 ⁇ -reductase activity was measured by analyzing the extent of the conversion of [ 3 H]testosterone to [ 3 H]5 ⁇ -DHT.
  • Microsomes are prepared at 4° C. from a buffered 0.32M-sucrose homogenate of human liver and from the livers of adult Sprague-Dawley female rats by differential centrifugation, and are used in the assay of 5 ⁇ -reductase activity. In some experiments, microsomes are solubilized with 0.1% polyoxyethylene ether W-1, except for the substitution of polyoxyethylene ether W-1 for Lubrolx-WX.
  • Cells genetically engineered to express specific types of 5 ⁇ -reductase isozymes can also be used as sources of 5 ⁇ -reductase activity.
  • Intact cells containing 5 ⁇ -reductase, their microsomes, or nuclear preparations can also be used to screen 5 ⁇ -reductase inhibitors.
  • a compound of this invention can be used to treat breast or prostate cancer.
  • the effectiveness of such compounds against prostate and breast cancer can be determined either on isolated cell lines derived from such cancer tissues or in animals demonstrating these forms of cancer.
  • human prostate cancer PC-3 cells are grown in culture medium. About one million cells are injected into male nude mice and the growth of tumors followed. Within two weeks, the tumor grows to about 100 mm 3 . Three tumor bearing mice are injected with a test compound each day.
  • a compound of this invention can be used to decrease organ and body weight.
  • the compounds thus have use in treating obesity.
  • the effectiveness of a compound can be determined using well-known animal models.
  • mice male Sprague-Dawley rats (body weight 180 g ⁇ 10 g) are used. Compounds are intraperitoneally injected into rats in one group each day for 7 days. Rats in the control group receive 0.1 ml 30% ethanol. Body and organ weights are determined.
  • inhibition of androgen action by topical administration of ⁇ -LA in hamster flank organs is investigated.
  • hamster flank organ Especially useful in the evaluation of the effects of these compounds on skin cells or sebaceous glands is the hamster flank organ (Frost and Gomez, 1972).
  • the paired flank organs, one on each side of the costovertebral angle, are highly sensitive to androgen stimulation.
  • the androgen sensitive structures in the flank organ include dermal melanocytes, sebaceous glands, and hair follicles (Hamilton and Montagna, 1950).
  • This animal model has been widely used for testing androgenic (Hamilton and Montagna, 1950; Frost et al., 1973) and antiandrogenic compounds (Voigt and Hsia, 1973; Weissmann et al., 1985; Chakrabarty et al., 1980).
  • the unique advantage of this animal model is that a testing compound can be applied topically to only one of the flank organs and the effect observed on both organs. If the test compound has only a local effect, then only the treated flank organ is affected. However, if the effect is systemic, then both flank organs are affected.
  • Pre-pubertal male Syrian golden hamsters castrated at 4 weeks old, are obtained from Harlan Sprague-Dawley Co. (Madison, Wis.). Each animal is maintained individually in a plastic cage on rodent chow (Purina) and water ad libitum on a 12 hour light/12 hour dark cycle.
  • a treatment solution (5 ⁇ l) is applied topically to the right flank organ once a day using a Pipetteman and a polypropylene disposable tip. Unless specified, the left flank organ is not treated.
  • the treatment solution contains either (a) ethanol alone (vehicle and control), or (b) a test compound. The flank organ was wiped with an alcohol pad to remove residual compound before each treatment.
  • the animals were sacrificed by either suffocation with CO 2 gas or with an intraperitoneal injection of an overdose of phenobarbital (64.8 mg/ml/animal).
  • the flank organs, both the treated and untreated sides, are evaluated to determine the effect of these treatments on the growth of the pigmented macule and the sebaceous glands.
  • the body weight of each animal is recorded before and after treatment.
  • the lengths of the long axis and the short axis of the pigmented spot are measured using a caliper with digital display (Digimatic, Mitutoyo Corp., Japan).
  • the product (long axis ⁇ short axis, mm 2 ) is used as an index of the surface area (Wuest and Lucky, 1989).
  • the flank organ treated with test compound becomes elevated and palpable.
  • the length of the long axis and short axis of the elevated mass are measured with a caliper.
  • the product of the long axis ⁇ short axis (mm 2 ) was used as an index of the areas of the sebaceous gland, which correlates with the volume of the sebaceous glands.
  • the stumptail macaque monkey develops baldness in a pattern resembling human androgenetic alopecia.
  • the balding process begins shortly after puberty (approximately 4 years of age). This occurs in nearly 100% of the animals, males and females, and is androgen dependent.
  • This is a useful animal model for human androgenetic alopecia and is contemplated to be useful in demonstrating the effects of polyunsaturated fatty acids on hair loss.
  • Male stumptail macaques (4 years of age) are divided into groups of 3 to 5 animals.
  • a defined area of the scalp involving the frontal and vertex areas is marked, e.g., by tattoo. Hairs in the marked area are shaved.
  • the solutions of a test compound in different dosages and combinations are evenly applied to the shaved areas once or twice a day.
  • Control animals receive the same volume of the solvent (e.g., ethanol or other organic solvent, or a cream).
  • the same area of the scalp is shaved every 4 to 6 weeks and the weights of hairs shaved are determined. The treatments may last for 6 months to 2 years.
  • Topical antiandrogenic activity of several fatty acids and catechins is first evaluated in the hamster flank organ assay or the rat assay. To further confirm the effectiveness of antiandrogenic compounds and suitability for human use, tests are performed on a human male subject.
  • the ideal compounds for human treatment are those that are topically and locally active but do not show systemic antiandrogenic activity, especially in the cases involving young males.
  • a male volunteer is used to test and analyze sebum production from the forehead region.
  • the forehead is washed thoroughly with soap twice and then cleaned with 70% isopropyl alcohol twice.
  • Sebum production is measured 30 to 60 minutes later with a sebum meter tape probe (7 mm ⁇ 8 mm) covering 56 mm 2 area in each measurement.
  • Ten measurements are made within the 4 cm square area (16 cm 2 ) located at the middle of the left or right side forehead between the eyebrow and the hair line.
  • the sebum meter detects the difference in the transparency of the tape before and after the tape was placed on the forehead for 30 seconds and expresses the difference in an arbitrary number (S-value) between 0 to 300 (or higher).
  • S-values of sebum accumulated on the foreheads of men are usually 200 to 300. Skin surface on hands usually shows a very low number (5 to 20).
  • the S-value for forehead immediately after washing is less than 5. For men, the S-value gradually increases to about 50 within 30 minutes after washing and reaches 100 to 200 in 45 minutes to 55 minutes.
  • the left and the right forehead areas are measured alternatively and each time at the comparable areas on the two sides. Ten measurements on each side (i.e., 20 measurements for two sides) take about 15-20 minutes and the sebum-values likely range between 30 to 200.
  • the S-values can differ considerably at different areas of the forehead and could be influenced by environmental, including weather, diet, and physiological, conditions.
  • the ratio of the total S-value (the sum of 10 measurements) for the left and the total S-value for the right forehead is constant. Therefore, compounds applied to the left forehead that reduce the L/R ratio to lower than 1.1 are considered as topically active agents for suppression of sebum production.
  • Aqueous compositions of the present invention comprise an effective amount of the 5 ⁇ -reductase inhibitory agent dissolved or dispersed in a pharmaceutically acceptable aqueous medium.
  • pharmaceutically acceptable refers to molecular entities and compositions that do not produce an allergic or similar untoward reaction when administered to a human.
  • compositions that contains such an inhibitory compound as an active ingredient are well understood in the art.
  • such compositions are prepared as injectable, either as liquid solutions or suspensions; solid forms suitable for solution in, or suspension in, liquid prior to injection can also be prepared.
  • the preparation can also be emulsified.
  • compositions disclosed herein may be orally administered, for example, with an inert diluent or with an assimilable edible carrier, or they may be enclosed in hard or soft shell gelatin capsule, or may be compressed into tablets, or they may be formulated for controlled release, such as a transdermic and osmotic pressure device, injectable device, and implantable device, or they may be incorporated directly with the food of the diet.
  • the active compounds may be incorporated with excipients and used in the form of ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, wafers, and the like.
  • the percentage of the compositions and preparations may, of course, be varied and may conveniently be 100% (application of pure compounds).
  • the amount of active compounds in such therapeutically useful compositions is such that a suitable dosage will be obtained.
  • the tablets, troches, pills, capsules and the like may also contain the following: a binder, such as gum tragacanth, acacia, corn starch or gelatin; excipients, such as dicalcium phosphate; a disintegrating agent, such as corn starch, potato starch, alginic acid and the like; a lubricant, such as magnesium stearate; and a sweetening agent, such as sucrose, lactose or saccharin; or a flavoring agent, such as peppermint, oil of wintergreen, or cherry flavoring.
  • a binder such as gum tragacanth, acacia, corn starch or gelatin
  • excipients such as dicalcium phosphate
  • a disintegrating agent such as corn starch, potato starch, alginic acid and the like
  • a lubricant such as magnesium stearate
  • a sweetening agent such as sucrose, lactose or saccharin
  • a flavoring agent such as
  • any material may be present as coatings or to otherwise modify the physical form of the dosage unit.
  • tablets, pills, or capsules may be coated with shellac, sugar or both.
  • a syrup of elixir may contain the active compounds; sucrose, as a sweetening agent, methyl and propylparabens as preservatives; a dye and flavoring, such as cherry or orange flavor.
  • any material used in preparing any dosage unit form should be pharmaceutically pure and substantially non-toxic.
  • the active compounds may be incorporated into sustained-release preparations and formulations.
  • the active compounds may also be administered parenterally, intravenously, or intraperitoneally.
  • Solutions of the active compounds as a free base or pharmacologically acceptable salts can be prepared in water suitably mixed with a surfactant, such as hydroxypropylcellulose.
  • Dispersions can also be prepared in glycerol, liquified polyethylene glycols, and mixtures thereof and in oils. Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms.
  • the pharmaceutical forms suitable for injectable use include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions.
  • the form must be sterile and must be fluid to the extent that easy syringability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms, such as bacteria and fungi.
  • the carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetable oils.
  • the proper fluidity can be maintained, for example, by the use of a coating, such a lecithin, by the maintenance of the required particle size in the case of a dispersion and by the use of surfactants.
  • a coating such as a lecithin
  • surfactants for example, sodium bicarbonate, sodium bicarbonate, sodium bicarbonate, sodium bicarbonate, sodium bicarbonate, sodium bicarbonate, sodium bicarbonate, sodium bicarbonate, sodium sorbic acid, thimerosal, and the like.
  • isotonic agents for example, sugars or sodium chloride.
  • Prolonged absorption of the injectable compositions can be brought about by the use in the compositions of agents delaying absorption, for example, aluminum monostearate and gelatin.
  • “pharmaceutically acceptable carrier” includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents and the like. The use of such media and agents for pharmaceutical active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active ingredient, its use in the therapeutic compositions is contemplated. Supplementary active ingredients can also be incorporated into the compositions.
  • the composition may be incorporated with excipients and used in the form of non-ingestible mouthwashes and dentifrices.
  • a mouthwash may be prepared incorporating the active ingredient in the required amount in an appropriate solvent, such as a sodium borate solution (Dobell's Solution).
  • the active ingredient may be incorporated into an antiseptic wash containing sodium borate, glycerin and potassium bicarbonate.
  • the active ingredient may also be dispersed in dentifrices, including: gels, pastes, powders and slurries.
  • the active ingredient may be added in a therapeutically effective amount to a paste dentifrice that may include water, binders, abrasives, flavoring agents, foaming agents, and humectants.
  • composition can be formulated in a neutral or salt form.
  • Pharmaceutically acceptable salts include the acid addition salts (formed with the free amino groups of the protein) and which are formed with inorganic acids such as, for example, hydrochloric or phosphoric acids, or such organic acids as acetic, oxalic, tartaric, mandelic, and the like. Salts formed with the free carboxyl groups can also be derived from inorganic bases such as, for example, sodium, potassium, ammonium, calcium, or ferric hydroxides, and such organic bases as isopropylamine, trimethylamine, histidine, procaine and the like.
  • solutions Upon formation, solutions will be administered in a manner compatible with the dosage formulation and in such a manner as it therapeutically effective.
  • the formulations are easily administered in a variety of dosage forms such as injectable solutions, drug release capsules and the like.
  • compositions disclosed herein may be formulated in creams, lotions, solutions, or in solid form depending upon the particular application.
  • the formulation of pharmaceutically acceptable vehicles for topical administration is well known to one of skill in the art (see, i.e., “Remington's Pharmaceuticals Sciences”, 15 th Edition). Variation of the dosage of the compositions disclosed herein, will necessarily depend upon the particular subject, and the nature of the condition(s) being treated.
  • aqueous solutions for parenteral administration in an aqueous solution, for example, the solution should be suitably buffered if necessary and the liquid diluent first rendered isotonic with sufficient saline or glucose.
  • aqueous solutions are especially suitable for intravenous, intramuscular, subcutaneous and intraperitoneal administration.
  • sterile aqueous media which can be employed will be known to those of skill in the art in light of the present disclosure.
  • one dosage could be dissolved in 1 ml of isotonic NaCl solution and either added to 1000 ml of hypodermic or intravenous fluid or injected at the proposed site of infusion, (see, for example, “Remington's Pharmaceutical Sciences”, 15 th Edition, pages 1035-1038 and 1570-1580). Some variation in dosage will necessarily occur depending on the condition of the subject being treated. The person responsible for administration will, in any event, determine the appropriate dose for the individual subject. Moreover, for human administration, preparations should meet sterility, pyrogenicity, general safety and purity standards as required by FDA Office of Biologics standards.
  • cDNAs for the human type 1 and 2 5 ⁇ -reductases were isolated from human prostate ⁇ gt11 and PC-3 cell ⁇ ZAP II cDNA libraries using the published sequence of the 5 ⁇ -reductases, PCR and standard library screening techniques.
  • the type 1 and 2 cDNAs were subcloned into the retroviral expression vector pMV7 and high titer stocks of virus containing the type 1 and 2 cDNAs were generated using the packaging cells BOSC 23 293.
  • Rat 1A cells were infected with virus and cells containing integrated retrovirus were selected for G418 resistance (Brown and Scott, 1987).
  • the tea catechins, ECG and EGCG had the highest activity of the tested flavanoids and were better inhibitors of the type 1 (HRED1) than the type 2 (HRED2) isoenzyme of 5 ⁇ -reductase.
  • the tea catechins epicatechin (EC) and epigallocatechin (EGC) had little activity.
  • Four flavanoids, myricetin, quercitin, baicalein and fisetin had significant (IC50 ⁇ 100 ⁇ M) activity and were more active against the type 1 than the type 2 isoenzyme.
  • Biochanin A, kaempferol, genistein, and diadzein were effective inhibitors of the type 2 but not type 1 isoenzyme.
  • Taxifolin a flavanone
  • the weak activity of taxifolin is most likely due to the absence of the 2,3-unsaturated bond when its activity is compared to the structurally related quercitin.
  • most flavanoids showed little or no activity against the type 1 isoenzyme, perhaps indicating limited penetration of these polyhydroxy compounds across the cell membrane.
  • four flavanoids, biochanin A, diadzein, kaempferol and genistein had significant inhibitory activity against the type 2 isoenzyme in the whole cell assay.
  • the most active of these, biochanin A and diadzein have only two and three free hydroxyl groups, respectively, and so may penetrate cells easier than other flavanoids.
  • Curcumin was a very effective inhibitor of either the type 1 or type 2 isoenzyme (Table 3, FIG. 3). TABLE 3 Cell Free Assay HRED1 HRED2 Isoenzyme HRED1 % Inhibition HRED2 % Inhibition Compound IC50(uM) @ 100 uM IC50(uM) @ 100 uM 1. Curcumin 3 95 5 87 2. Tetrahydro- 80 56 29 73 curcumin 3. Demethoxy- >100 23 >100 42 tetrahydrocurcumin 4. 4-hydroxy-3- >100 10 >100 ( ⁇ 60) methoxy- cinnamaldehyde 5. Coniferol >100 10 100 49 6. 4-(4-hydroxy-3- >100 3 >100 4 methoxyphenol)-3- buten-2-one 7. Ferulic Acid >100 0 >100 18 8. Capsaicin >100 0 >100 8 9. Eugenol >100 0 100 50
  • alizarin The naturally occurring anthraquinone, alizarin, was a very effective inhibitor of the type 1 but not type 2 isozymes.
  • Alizarin Red S which is a water soluble sulfate derivative of alizarin had little activity (IC50s>100 ⁇ M) against either isoenzyme.
  • the charged sulfate group may prevent interaction with membrane bound 5 ⁇ -reductase.
  • Purpurin which has an additional hydroxyl compared to alizarin, had inhibitory activity similar to alizarin.
  • Anthraflavic acid, anthrarufin and quinizarin which are structural isomers of alizarin without adjacent hydroxyl groups, had much less activity, emphasizing the importance of the catechol moiety for potent inhibitory activity of this class of anthroquinones.
  • Anthraquinone was not an effective inhibitor (IC50>100 ⁇ M).
  • Menadione, coenzyme Q, and 2,6-dichloroindophenol were potent cell-free inhibitors of boh isoenzymes. The compounds participate in quinone reductase reactions and may deplete NADPH causing the observed inhibition.
  • alizarin was a very effective inhibitor of the type 1 isoenzyme and menadione had moderate activity.
  • HZIV 142 25 97 63 93 7. HZIV 68 29 93 99 51 8. HZIV 75 29 97 >100 21 9. HZIV 166 30 98 78 74 10. HZIV 63 311 94 >100 20 11. HZIV 169 47 90 >100 39 12. HZIV 74 48 85 >100 24 13. HZIV 144 49 88 >100 38 14. HZIV 168 49 98 73 92 15. HZIV 166 59 95 71 84 16. BGC 62 61 >100 30 17. HZIV 107 98 52 >100 39 18. HZIV 145 >100 35 >100 8 19. HZIV 148 >100 31 >100 0 20. HZIV 109 >100 17 >100 0
  • Alpha-Linolenic Acid 8 99 9 84 C18:3 CIS 9,12,15 4.
  • Linoleic Acid 9 99 19 85 C18:2 CIS 9,12 5.
  • Conjugated 10 99 30 81 Octadecadienonic Acid 7. 5,8,11,14-Eocpsatetraynoic Acid 15 97 3 81 8.
  • EGCG >100 11 >100 5 27. Myricetin >100 11 >100 11 28. Purpurin >100 47 >100 7 29. Quercetin >100 15 >100 29 30. Alizarin Red S >100 28 >100 1 31. Genistein >100 22 20 89 32. HZIV 123 >100 48 >100 8 33. HZIV 107 >100 23 >100 2 34. Catechol >100 9 >100 3 35. Daidzein >100 9 58 87 36. Pyrogallol >100 7 >100 15 37. EC >100 0 >100 1 38. EGC >100 15 >100 1 39. ECG >100 0 >100 0 40. EGCG >100 6 >100 0 41. HZIV 90 >100 34 >100 14 42.
  • HZIV 63 >100 12 >100 7 43.
  • HZIV 68 >100 40 >100 34 44.
  • HZIV 144 >100 12 >100 7 45.
  • HZIV 81-3 >100 28 19 80 46.
  • HZIV 145 >100 8 >100 9 47.
  • Methyl Gallate >100 0 >100 0 48.
  • Propyl Gallate >100 5 >100 0 49.
  • Gallic Acid >100 13 >100 0 51.
  • Pyrogallol >100 5 >100 6 52.
  • HZIV 169 >100 10 >100 0 53.
  • the naturally occurring fatty acid, ⁇ -linolenic acid has IC50 of 3 ⁇ M for both isoenzymes.
  • Fatty acids such as ETYA may be useful for derivatizing other 5 ⁇ -reductase inhibitors to enhance cellular uptake and promote in vivo activity of 5 ⁇ -reductase inhibitors.
  • Methyl and cholesterol esters of ⁇ -linolenic acid had little activity in the whole cell assay (TABLE 7) and so the activity of EGC esterified to ⁇ -linolenic acid is unlikely due to intracellular hydrolysis of these esters.
  • Active 5 ⁇ -reductase inhibitors shown in Tables 1-7 are polyphenols or their derivatives and are easily oxidized or hydrolyzed within several hours to several days, especially in the presence of air or oxygen and at a pH above 7.0. We have found that these compounds are more stable to oxidation or hydrolysis by maintaining the pH of the solutions of these compounds at a pH below 7.0. More than 80% of the oxidation of hydrolysis can be prevented by the addition of an inorganic acid, such as hydrochloric acid, sulfuric acid, or phosphoric acid, or an organic acid, such as citric acid or acetic acid.
  • an inorganic acid such as hydrochloric acid, sulfuric acid, or phosphoric acid
  • an organic acid such as citric acid or acetic acid.

Abstract

Compounds that inhibit 5α-reductase are provided. The compounds are used to treat prostate cancer, breast cancer, obesity, skin disorders and baldness.

Description

    TECHNICAL FIELD OF THE INVENTION
  • The present invention relates generally to compounds, compositions and methods regulating the action and function of androgens and other steroid hormones by modulating the activity of steroid-reductases, including isozymes of 5α-reductases. More specifically, the present invention relates to the use of these compounds to regulate processes or treat disorders that are modulated by androgens or other steroid hormones or are caused by abnormal actions of androgens or other steroid hormones in cells or organs of animals, humans, plants, or microorganisms. This invention relates to the use of natural and synthetic flavanoids, catechols, curcumin-related substances, quinones, catechins and fatty acids and their analogues or derivatives as 5α-reductase isozyme inhibitors and as therapeutic agents. These compounds can also be used in promoting or modulating desirable production of specific products for commercial purposes. [0001]
  • BACKGROUND OF THE INVENTION
  • In some of the androgen-sensitive organs, such as the prostate and skin, testosterone (T) is converted to a more active metabolite 5α-dihydrotestosterone (DHT) by 5α-reductase (Anderson and Liao, 1968; Bruchovsky and Wilson, 1968). Other substrates of 5α-reductases are also converted to reduce products that may have specific properties. Inhibition of 5α-reductase represents a unique approach for developing therapeutic methods for androgen-dependent diseases, such as benign prostatic hyperplasia, breast and prostatic cancer, skin disorders, seborrhea, common baldness, hirsutism, and hidradenitis suppurative. Various compounds have been shown to inhibit 5α-reductase activity (Liang and Liao, 1992; Hirsch et al., 1993; Russell and Wilson, 1994; Liao and Hiipakka, 1995). Finasteride (Proscar), a 5α-reductase inhibitor, lowers the level of DHT in serum and the prostate, reduces prostate volume and increase urinary flow in some patients (Stoner E. Finasteride Study Group, 1992). Certain aliphatic unsaturated fatty acids, such as γ-linolenic acid (Liang and Liao, 1992) and catechin-3-gallates (Liao and Hiipakka, 1995), can inhibit 5α-reductase activity of liver and prostate of rats and humans in vitro. [0002]
  • 5α-Reductase is found in many organs (Russell and Wilson, 1994; Hiipakka et al., 1993) including the sebaceous gland of hamsters (Takayasu and Adachi, 1972) and human hair follicles (Randall, 1994). Two 5α-reductase isozymes have been identified in rats and humans (Russell and Wilson, 1994). The [0003] type 1 isozyme predominates in rat tissues such as liver, kidney, brain, and lung, whereas the type 2 enzyme is more abundant in rat testis and epididymis. Both isozymes are found in skins of the neonate, but the type 1 isozyme is the major form expressed in the skin after puberty. The type 1 isozyme is also expressed in balding scalp. The possibility that the type 2 isozyme plays a unique role in skin and hair growth cannot be excluded. Finasteride, a 4-azasteroid, is a competitive inhibitor of 5α-reductases and has an affinity 30-fold higher for isozyme 2 than for isozyme 1 (Russell and Wilson, 1994). In contrast, the green tea catechins, epicatechin-3-gallate and epigallocatechin-3-gallate are more effective inhibitors of the type 1 enzyme and γ-linolenic acid inhibits both isozymes equally well (Liao and Hiipakka, 1995).
  • In the stumptail macaque, a monkey model of androgenic alopecia, finasteride given orally prevents frontal baldness (Diani et al, 1992). The paired hamster flank organs, one on each side of the costovertebral angle, are highly sensitive to androgen stimulation. Topical application of γ-linolenic acid suppresses only the androgen-dependent growth of the treated hamster flank organ without showing systemic effects on the contralateral flank organ and this effect is very likely due to local inhibition of 5α-reductase. [0004]
  • Uses of androgens known to the medical arts include, for example, treatment of hypogonadism and anemia. The abuse of androgens among athletes to enhance performance is well known. Androgens are also known to promote the development of benign prostatic hyperplasia (BPH), prostate cancer, baldness, acne, obesity and undesirable lipid and steroid profiles in blood and organs. Approximately 70% of males in the U.S. over the age of 50 have pathological evidence of BPH. Prostate cancer is the second leading cause of cancer death in males in the U.S. Male-patterned baldness can start as early as the teens in genetically susceptible males, and it has been estimated to be present in 30% of Caucasian males at age 30, 40% of Caucasian males at age 40, and 50% of Caucasian males at age 50. Acne is the most common skin disorder treated by physicians. In women, hirsutism is one of the hallmarks of excessive androgen. The ovaries and the adrenal are the major sources of androgen in women. [0005]
  • In men, the major androgen circulating in the blood is testosterone. About 98% of the testosterone in blood is bound to serum proteins (high affinity finding to sex-steroid binding globulin and low affinity binding to albumin), with only 1-2% in free form. The albumin-bound testosterone, the binding of which is readily reversible, and the free form are considered to be bioavailable, and account for about 50% of total testosterone. Testosterone enters target cells apparently by diffusion. In the prostate, seminal vesicles, skin, and some other target organs, it is converted by a NADPH-dependent 5α-reductase to a more active metabolite, 5α-DHT. 5α-DHT then binds an androgen receptor (AR) in target organs. The 5α-DHT-receptor complexes interact with specific portions of the genome to regulate gene activities (Liao et al., 1989). Testosterone appears to bind to the same AR, but it has a lower affinity than 5α-DHT. In tissues such as muscle and testes, where 5α-reductase activity is low, testosterone may be the more active androgen. [0006]
  • The difference between testosterone and 5α-DHT activity in different androgen-responsive tissues is further suggested by findings in patients with 5α-reductase deficiency. Males with 5α-reductase deficiency are born with female-like external genitalia. When they reach puberty, their plasma levels of testosterone are normal or slightly elevated. Their muscle growth accelerates, the penis enlarges, voice deepens, and libido toward females develops. However, their prostates remain non-palpable, they have reduced body hair, and they do not develop acne or baldness. [0007]
  • The findings in 5α-reductase deficient patients suggest that inhibitors of 5α-reductase would be useful for the treatment of prostatic cancer, BPH, acne, baldness, and female hirsutism. Clinical observations and animal experiments have indicated that spermatogenesis, maintenance of libido, sexual behavior, and feedback inhibition of gonadotropin secretion do not require the conversion of testosterone to 5α-DHT. This is in contrast to other hormonal therapies which abolish the actions of both testosterone and 5α-DHT. [0008]
  • Treatment of androgen-dependent skin and prostatic diseases by 5α-reductase inhibitors would be expected to produce fewer side effects than the presently available hormonal therapies. These include castration, estrogen therapy, high doses of superactive gonadotropin-releasing hormone such as Luprolide, and the use of competitive antiandrogens which inhibit AR binding of testosterone and 5α-DHT, such as flutamide, cyproterone acetate and spironolactone. The long term efficacy of competitive antiandrogens is also compromised by their block of the androgenic feedback inhibition of gonadotropin secretion. This increases testicular secretion of testosterone. The higher level of testosterone eventually overcomes the action of the antiandrogen. [0009]
  • Excessive 5α-DHT is implicated in certain androgen-dependent pathological conditions including BPH, acne, male-pattern baldness, and female idiopathic hirsutism. It has been shown that 5α-reductase activity is reported to be higher in hair follicles from the scalp of balding men than that of non-balding men. [0010]
  • Since treatments of androgen-dependent skin and prostatic diseases by 5α-reductase inhibitors can produce fewer side effects than the hormonal therapies which indiscriminately inhibit all androgen actions, it is desirable to provide different types of 5α-reductase inhibitors. [0011]
  • BRIEF SUMMARY OF THE INVENTION
  • The present invention relates generally to the utilization of certain compounds for the control of androgen activity in target organs and cells through the modulation of a 5α-reductase activity. In certain aspects, these compounds are employed to repress androgenic activity by inhibiting the formation and availability of active androgen in target cells. Consequently, the present invention is useful for the treatment of a wide variety of conditions including, but not limited to, the treatment of prostatic hyperplasia, prostatic cancer, hirsutism, acne, male pattern baldness, seborrhea, and other diseases related to androgen hyperactivity. Several of these compounds have been shown to effectively decrease body weight, and in some cases, to decrease the weight of an androgen-dependent body organ, such as the prostate and other organs. The effectiveness of these compounds may be dependent also on their action on other mechanisms involved in angiogenesis, cell-cell interaction, and on their interaction with various components of organs and cells. [0012]
  • Compounds useful in the practice of the present invention include various isomers of saturated and unsaturated fatty acids, natural and synthetic analogues, and derivatives from which these fatty acids can be generated as well as the metabolites and oxidation products of these fatty acids. The use of these and other fatty acids and their derivatives is also contemplated. Also useful are catechin compounds, particularly, catechins that are structurally similar to epicatechin gallate (ECG) and epigallocatechin gallate (EGCG). EGCG has an additional hydroxyl group on the epicatechin gallate molecule, which has been found to be surprisingly active in modulating several 5α-reductase mediated processes. EGCG derivatives having such an additional OH group on the altering ECG molecule were shown to be active in inducing body weight loss and particularly in reducing the size of androgen sensitive organs such as preputial glands, ventral prostate, dorsolateral prostate, coagulating glands, seminal vesicles, human prostate tumors, and breast tumors in nude mice. [0013]
  • By analogy with the fatty acid compounds, certain active catechin gallates may not enter target cells easily. However, esterification of hydroxyl groups on the inhibitor compounds should enhance the ability of these compounds to enter the target cells. Once inside the cells, esters would be readily hydrolyzed by esterase to alcohols that can inhibit 5α-reductases (Williams, 1985). [0014]
  • In more particular aspects of the invention, the inventors have discovered that certain catechins, particularly EGCG, can be administered to promote body weight loss that differentially affects overall body weight and prostate weight loss. In particular examples, it was shown that for a certain percentage of overall body weight loss, prostate weight loss was percentage-wise more than three times as much. The loss in body weight and the organ weight are likely due to EGCG interference of a common step in the pathway controlling body weight and the organ weight gain. EGCG and related compounds may interact and interfere with a receptor macromolecule (probably containing a protein) that modulates specific lipid synthesis and accumulation. Lipids can modulate gene expression, cell development and differentiation, and organ growth. Specific interference of lipid metabolism in the cells and organs may control the growth of the organs, in particular, prostate, sebaceous, preputial and other secretory organs. In certain applications, it is expected that benign or abnormal growth or cancer of these organs may be treated or even prevented by administration of catechin related compounds. [0015]
  • It has been demonstrated that catechin compounds will arrest or reduce human prostate and breast cancer cell growth. The effectiveness of catechin compounds was shown to be dependent on the methods by which these compounds were administered to the experimental animals. Intraperitoneal application was much more effective than oral administration. It is expected that direct application to the organs, such as the prostate, will be very effective. EGCG was surprisingly effective in suppressing and even reducing the size of human prostate and breast tumors in animal models. The effect was illustrated with EGCG; however, structurally similar catechin compounds are also effective, particularly those that are structurally similar to EGCG in having at least one additional hydroxyl group as compared with ECG. Thus, the EGCG species that contain eight hydroxyl groups is significantly more effective in reducing body weight than is ECG, which contains seven hydroxyl groups. Compounds of this general structure are expected to be particularly effective in chemoprevention and chemotherapy of human prostate cancer. Compounds having a part of structure similar to a part of structure of EGCG are also expected to be effective. [0016]
  • Compounds can be used as antiandrogenic agents through topical or systemic application. A preparation for this purpose can include a carrier, a protectant, an antioxidant (such as vitamin C or E, and various catechins and polyphenols), and other pharmaceutical and pharmacological agents. It is also expected that such compounds can be used in a delivery system (oral, local application, injection, or implantation) involving molecular recognition through which the compounds are delivered to target sites. Such a delivery system may involve, among other methods, liposome techniques or immunological devices. [0017]
  • Natural or synthetic chemicals that can modulate the production or cellular action of receptors and macromolecules are useful in the treatment of abnormalities such as obesity, BPH, prostate cancer, skin diseases, baldness, breast tumors, and hirsutism, which are related to lipid synthesis, body weight, and/or androgen function. [0018]
  • Animal models can be used to demonstrate the effectiveness of compounds on a variety of cancers. For example, Shionogi tumor and other tumors can be studied in male rats. Human breast and prostate cancer cell growth can be studied in nude mice. Alternatively, rodent breast tumors induced by carcinogens and other cancers induced in transgenic mice or Dunning tumor in rats can be similarly analyzed for their chemotherapy by EGCG and related compounds. [0019]
  • The use of compounds disclosed in the present invention, or in natural therapeutically effective amounts of pharmaceutical compositions containing one or more of the compounds, in some cases in combination with other therapeutic agents and carriers, or in natural or synthetic products, is appropriate in the treatment of various disorders. These disorders include, but are not necessarily limited to, those conditions wherein excessive androgenic activities have been implicated, for example, male pattern baldness, female hirsutism, skin disorders, BPH, cancers of prostate, breast, skin and other organs. [0020]
  • The present invention is also directed to novel compounds. These compounds have the formula: [0021]
    Figure US20030144346A1-20030731-C00001
  • where x is —NHCH[0022] 2CH2— or —CH═CH—;
  • R[0023] 1, R2 and R3 each may be —H, —OH or —OCH3, provided that only one of R1, R2, and R3 may be —H;
  • R[0024] 4, R5 and R6 each may be —H, —OH, —OCH3 or —N(CH3)2, provided that only one of R4, R5 and R6 may be —H; and
  • n is 0 or 1. [0025]
  • and the formula: [0026]
    Figure US20030144346A1-20030731-C00002
  • where R is [0027]
    Figure US20030144346A1-20030731-C00003
  • All of the compositions and methods disclosed and claimed herein can be made without undue experimentation in light of the present disclosure. While the compositions and methods of this invention have been described in terms of preferred embodiments, it will be apparent to those of skill in the art that variations may be applied to the compositions, methods and in the steps or in the sequence of steps of the method described herein without departing from the concept, spirit and scope of the invention. More specifically, it will be apparent that certain agents which are both chemically and physiologically related may be substituted for the agents described herein while the same or similar results would be achieved. All such similar substitutes and modifications apparent to those skilled in the art are deemed to be within the spirit, scope and concept of the invention as defined by the appended claims.[0028]
  • BRIEF DESCRIPTION OF THE DRAWINGS
  • In the drawings, which form a portion of the specification: [0029]
  • FIG. 1 shows the structure of flavanoid compounds of the present invention. [0030]
  • FIG. 2 shows the structure of catechol compounds of the present invention. [0031]
  • FIG. 3 shows the structure of curcumin and related compounds of the present invention. [0032]
  • FIG. 4. shows quinones of the present invention. [0033]
  • FIG. 5 shows epigallocatechin derivative compounds of the present invention. [0034]
  • FIG. 6 shows the generic formula of the epigallocatechin derivatives of the present invention; [0035]
  • FIG. 7 shows the generic formula of gallates useful in the present invention; [0036]
  • FIG. 8 shows the generic formula of curcumin derivatives useful in the present invention; [0037]
  • FIG. 9 shows the generic formula of quinones and catechols useful in the present invention. [0038]
  • FIG. 10 shows fatty acids of the present invention. [0039]
  • DETAILED DESCRIPTION OF THE INVENTION
  • I. 5α-Reductase Activity [0040]
  • The present invention is concerned with methods of inhibiting 5α-reductase, which include subjecting a cell to an effective concentration of a 5α-reductase inhibitor such as one of the compounds disclosed herein. It is believed that the use of such inhibitors to block abnormal androgen action will serve to treat cancer in conjunction with other anti-cancer agents, chemotherapy, resection, radiation therapy, and the like. The compounds of this invention, besides acting as 5α-reductase inhibitors, may have other effects that can lead to antitumor activity or to suppress abnormal growth of prostate or other organs. [0041]
  • In mammalian cells, 5α-reductase is very tightly associated with intracellular membranes, including the membrane of the endoplasmic reticulum and contiguous nuclear membranes. Attempts to solubilize and purify active 5α-reductase have not been very successful. The assay of 5α-reductase activity, therefore, is performed by measuring the rate of conversion of testosterone to 5α-DHT by whole cells or by microsomal and nuclear preparations in the presence of NADPH (enzymatic assay). Alternatively, the 5α-reductase activity can be reliably assayed by following NADPH-dependent noncovalent binding of a potent radioactive inhibitor, such as [[0042] 3H]4-MA ([3H]4-MA-binding assay), which strongly competes with testosterone for binding to the reductase. The results of the two assays correlate very well when microsomal preparations from different organs or animals are used for comparison.
  • A. [[0043] 3H]4-MA Binding Assay For 5α-Reductase
  • Briefly, the binding assay solution, in a final volume of 0.15 ml, contains microsomes (2-20 μg of protein), 0.07 μCi of [[0044] 3H]4-MA, 0.1 mM-NADPH, 1 mM-dithiothreitol and 50 mM-potassium phosphate, pH 7.0, with or without the indicated amount of a lipid or an inhibitor preparation. Lipids are dissolved in ethanol and added in 1-5 μl volumes. Control tubes receive the same amount of ethanol. After incubation at 0° C. for 1 hour, the [3H]4-MA bound to microsomes is determined by collecting microsomes on a Whatman glass fibre filter and washing with 10 ml of 20 mM-potassium phosphate, pH 7.0, containing 0.01% CHAPS to remove unbound [3H]4-MA.
  • B. Assays Of The Enzymatic Activity Of Microsomal 5α-Reductase [0045]
  • The standard reaction mixture, in a final volume of 0.15 ml, contains microsomes, 1 μCi of [[0046] 3H]testosterone, 0.5-3.0 μM non-radioactive testosterone, 0.1 mM-NADPH, 1 mM-dithiothreitol and 50 mM-potassium phosphate, pH 7.0, with or without the indicated amount of a lipid or an inhibitor preparation. The reaction is started by the addition of microsomes and the incubation is carried out at 37° C for 15 minutes. Steroids are extracted and separated by thin layer chromatography. Radioactive steroids are located by fluorography and the amount of radioactivity present determined by scintillation counting. The 5α-reductase activity was measured by analyzing the extent of the conversion of [3H]testosterone to [3 H]5α-DHT.
  • C. Sources of 5α-Reductase Activity [0047]
  • Microsomes are prepared at 4° C. from a buffered 0.32M-sucrose homogenate of human liver and from the livers of adult Sprague-Dawley female rats by differential centrifugation, and are used in the assay of 5α-reductase activity. In some experiments, microsomes are solubilized with 0.1% polyoxyethylene ether W-1, except for the substitution of polyoxyethylene ether W-1 for Lubrolx-WX. [0048]
  • Cells genetically engineered to express specific types of 5α-reductase isozymes can also be used as sources of 5α-reductase activity. Intact cells containing 5α-reductase, their microsomes, or nuclear preparations can also be used to screen 5α-reductase inhibitors. [0049]
  • II. Prostate And Breast Cancer [0050]
  • A compound of this invention can be used to treat breast or prostate cancer. The effectiveness of such compounds against prostate and breast cancer can be determined either on isolated cell lines derived from such cancer tissues or in animals demonstrating these forms of cancer. By way of example, human prostate cancer PC-3 cells are grown in culture medium. About one million cells are injected into male nude mice and the growth of tumors followed. Within two weeks, the tumor grows to about 100 mm[0051] 3. Three tumor bearing mice are injected with a test compound each day.
  • III. Organ And Body Weight Loss [0052]
  • A compound of this invention can be used to decrease organ and body weight. The compounds thus have use in treating obesity. The effectiveness of a compound can be determined using well-known animal models. [0053]
  • By way of example, male Sprague-Dawley rats (body weight 180 g±10 g) are used. Compounds are intraperitoneally injected into rats in one group each day for 7 days. Rats in the control group receive 0.1 ml 30% ethanol. Body and organ weights are determined. [0054]
  • IV. Skin Disorders [0055]
  • The inventors sought an inhibitor of 5α-reductase that would be active topically and inactive systemically; such an agent would be ideal for treatment of androgen-dependent dermatological disorders. In this study, inhibition of androgen action by topical administration of γ-LA in hamster flank organs is investigated. Especially useful in the evaluation of the effects of these compounds on skin cells or sebaceous glands is the hamster flank organ (Frost and Gomez, 1972). The paired flank organs, one on each side of the costovertebral angle, are highly sensitive to androgen stimulation. The androgen sensitive structures in the flank organ include dermal melanocytes, sebaceous glands, and hair follicles (Hamilton and Montagna, 1950). This animal model has been widely used for testing androgenic (Hamilton and Montagna, 1950; Frost et al., 1973) and antiandrogenic compounds (Voigt and Hsia, 1973; Weissmann et al., 1985; Chakrabarty et al., 1980). The unique advantage of this animal model is that a testing compound can be applied topically to only one of the flank organs and the effect observed on both organs. If the test compound has only a local effect, then only the treated flank organ is affected. However, if the effect is systemic, then both flank organs are affected. [0056]
  • Pre-pubertal male Syrian golden hamsters, castrated at 4 weeks old, are obtained from Harlan Sprague-Dawley Co. (Madison, Wis.). Each animal is maintained individually in a plastic cage on rodent chow (Purina) and water ad libitum on a 12 hour light/12 hour dark cycle. [0057]
  • One to two weeks after castration, the hair on the lower back of each animal is clipped with an electric hair clipper and then shaved weekly to expose the flank organs. The animals are divided into treatment groups. A treatment solution (5 μl) is applied topically to the right flank organ once a day using a Pipetteman and a polypropylene disposable tip. Unless specified, the left flank organ is not treated. The treatment solution contains either (a) ethanol alone (vehicle and control), or (b) a test compound. The flank organ was wiped with an alcohol pad to remove residual compound before each treatment. At the end of each experiment (17-25 days), the animals were sacrificed by either suffocation with CO[0058] 2 gas or with an intraperitoneal injection of an overdose of phenobarbital (64.8 mg/ml/animal). The flank organs, both the treated and untreated sides, are evaluated to determine the effect of these treatments on the growth of the pigmented macule and the sebaceous glands. The body weight of each animal is recorded before and after treatment.
  • Treatment Of Animals [0059]
  • Male hamsters, 4 weeks old, are castrated and are kept on a longer light period (16 hours light/8 hours dark cycle) to insure maximum stimulation of sexual characteristics (Luderschmidt et al., 1984). Flank organs, left and right, were treated topically with 5 μl ethanol containing 0.5 μg or 1 μg testosterone daily. Animals are divided into groups of 4-5 hamsters. The right flank organ is also treated daily with 5 μl solution containing vehicle (ethanol) alone or test compound (1 or 2 mg) for 18 days. The left flank organ of all animals receives the same volume of vehicle. [0060]
  • the lengths of the long axis and the short axis of the pigmented spot (pigmented macule) are measured using a caliper with digital display (Digimatic, Mitutoyo Corp., Japan). The product (long axis×short axis, mm[0061] 2) is used as an index of the surface area (Wuest and Lucky, 1989).
  • The flank organ treated with test compound becomes elevated and palpable. The length of the long axis and short axis of the elevated mass are measured with a caliper. The product of the long axis×short axis (mm[0062] 2) was used as an index of the areas of the sebaceous gland, which correlates with the volume of the sebaceous glands.
  • V. Baldness [0063]
  • Topical Effects Of Compounds On Hair Loss And Growth [0064]
  • The stumptail macaque monkey develops baldness in a pattern resembling human androgenetic alopecia. The balding process begins shortly after puberty (approximately 4 years of age). This occurs in nearly 100% of the animals, males and females, and is androgen dependent. This is a useful animal model for human androgenetic alopecia and is contemplated to be useful in demonstrating the effects of polyunsaturated fatty acids on hair loss. The following describes a protocol for testing. [0065]
  • Male stumptail macaques (4 years of age) are divided into groups of 3 to 5 animals. A defined area of the scalp involving the frontal and vertex areas is marked, e.g., by tattoo. Hairs in the marked area are shaved. The solutions of a test compound in different dosages and combinations are evenly applied to the shaved areas once or twice a day. Control animals receive the same volume of the solvent (e.g., ethanol or other organic solvent, or a cream). The same area of the scalp is shaved every 4 to 6 weeks and the weights of hairs shaved are determined. The treatments may last for 6 months to 2 years. 4-MA (17-N,N-diethylcarbamoyl-4-methyl-4-aza-5-androstan-3-one), a 5α-reductase inhibitor known to prevent baldness in this animal is included as a positive control. Biopsies of the scalp (4 mm punch) are obtained before and at the end of the treatments. The specimens are analyzed for 5α-reductase activity and examined histologically for evidence of alopecia. [0066]
  • VI. Effects Of Compounds On Sebum Productions In A Human Model [0067]
  • Topical antiandrogenic activity of several fatty acids and catechins is first evaluated in the hamster flank organ assay or the rat assay. To further confirm the effectiveness of antiandrogenic compounds and suitability for human use, tests are performed on a human male subject. The ideal compounds for human treatment are those that are topically and locally active but do not show systemic antiandrogenic activity, especially in the cases involving young males. [0068]
  • Determination Of Forehead Sebum Production [0069]
  • A male volunteer is used to test and analyze sebum production from the forehead region. The forehead is washed thoroughly with soap twice and then cleaned with 70% isopropyl alcohol twice. Sebum production is measured 30 to 60 minutes later with a sebum meter tape probe (7 mm×8 mm) covering 56 mm[0070] 2 area in each measurement. Ten measurements are made within the 4 cm square area (16 cm2) located at the middle of the left or right side forehead between the eyebrow and the hair line.
  • The sebum meter detects the difference in the transparency of the tape before and after the tape was placed on the forehead for 30 seconds and expresses the difference in an arbitrary number (S-value) between 0 to 300 (or higher). S-values of sebum accumulated on the foreheads of men are usually 200 to 300. Skin surface on hands usually shows a very low number (5 to 20). The S-value for forehead immediately after washing is less than 5. For men, the S-value gradually increases to about 50 within 30 minutes after washing and reaches 100 to 200 in 45 minutes to 55 minutes. [0071]
  • To determine the rate of sebum production, the left and the right forehead areas are measured alternatively and each time at the comparable areas on the two sides. Ten measurements on each side (i.e., 20 measurements for two sides) take about 15-20 minutes and the sebum-values likely range between 30 to 200. The S-values can differ considerably at different areas of the forehead and could be influenced by environmental, including weather, diet, and physiological, conditions. However, the ratio of the total S-value (the sum of 10 measurements) for the left and the total S-value for the right forehead is constant. Therefore, compounds applied to the left forehead that reduce the L/R ratio to lower than 1.1 are considered as topically active agents for suppression of sebum production. [0072]
  • VII. Pharmaceutical Compositions [0073]
  • Aqueous compositions of the present invention comprise an effective amount of the 5α-reductase inhibitory agent dissolved or dispersed in a pharmaceutically acceptable aqueous medium. The phrase “pharmaceutically acceptable” refers to molecular entities and compositions that do not produce an allergic or similar untoward reaction when administered to a human. [0074]
  • The preparation of an aqueous composition that contains such an inhibitory compound as an active ingredient is well understood in the art. Typically, such compositions are prepared as injectable, either as liquid solutions or suspensions; solid forms suitable for solution in, or suspension in, liquid prior to injection can also be prepared. The preparation can also be emulsified. [0075]
  • The pharmaceutical compositions disclosed herein may be orally administered, for example, with an inert diluent or with an assimilable edible carrier, or they may be enclosed in hard or soft shell gelatin capsule, or may be compressed into tablets, or they may be formulated for controlled release, such as a transdermic and osmotic pressure device, injectable device, and implantable device, or they may be incorporated directly with the food of the diet. For oral therapeutic administration, the active compounds may be incorporated with excipients and used in the form of ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, wafers, and the like. The percentage of the compositions and preparations may, of course, be varied and may conveniently be 100% (application of pure compounds). The amount of active compounds in such therapeutically useful compositions is such that a suitable dosage will be obtained. [0076]
  • The tablets, troches, pills, capsules and the like may also contain the following: a binder, such as gum tragacanth, acacia, corn starch or gelatin; excipients, such as dicalcium phosphate; a disintegrating agent, such as corn starch, potato starch, alginic acid and the like; a lubricant, such as magnesium stearate; and a sweetening agent, such as sucrose, lactose or saccharin; or a flavoring agent, such as peppermint, oil of wintergreen, or cherry flavoring. When the dosage unit form is a capsule, it may contain, in addition to materials of the above type, a liquid carrier. Various other materials may be present as coatings or to otherwise modify the physical form of the dosage unit. For instance, tablets, pills, or capsules may be coated with shellac, sugar or both. A syrup of elixir may contain the active compounds; sucrose, as a sweetening agent, methyl and propylparabens as preservatives; a dye and flavoring, such as cherry or orange flavor. Of course, any material used in preparing any dosage unit form should be pharmaceutically pure and substantially non-toxic. In addition, the active compounds may be incorporated into sustained-release preparations and formulations. [0077]
  • The active compounds may also be administered parenterally, intravenously, or intraperitoneally. Solutions of the active compounds as a free base or pharmacologically acceptable salts can be prepared in water suitably mixed with a surfactant, such as hydroxypropylcellulose. Dispersions can also be prepared in glycerol, liquified polyethylene glycols, and mixtures thereof and in oils. Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms. [0078]
  • The pharmaceutical forms suitable for injectable use include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. In all cases, the form must be sterile and must be fluid to the extent that easy syringability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms, such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetable oils. The proper fluidity can be maintained, for example, by the use of a coating, such a lecithin, by the maintenance of the required particle size in the case of a dispersion and by the use of surfactants. The prevention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars or sodium chloride. Prolonged absorption of the injectable compositions can be brought about by the use in the compositions of agents delaying absorption, for example, aluminum monostearate and gelatin. [0079]
  • As used herein, “pharmaceutically acceptable carrier” includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents and the like. The use of such media and agents for pharmaceutical active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active ingredient, its use in the therapeutic compositions is contemplated. Supplementary active ingredients can also be incorporated into the compositions. [0080]
  • For oral administration the composition may be incorporated with excipients and used in the form of non-ingestible mouthwashes and dentifrices. A mouthwash may be prepared incorporating the active ingredient in the required amount in an appropriate solvent, such as a sodium borate solution (Dobell's Solution). Alternatively, the active ingredient may be incorporated into an antiseptic wash containing sodium borate, glycerin and potassium bicarbonate. The active ingredient may also be dispersed in dentifrices, including: gels, pastes, powders and slurries. The active ingredient may be added in a therapeutically effective amount to a paste dentifrice that may include water, binders, abrasives, flavoring agents, foaming agents, and humectants. [0081]
  • The composition can be formulated in a neutral or salt form. Pharmaceutically acceptable salts, include the acid addition salts (formed with the free amino groups of the protein) and which are formed with inorganic acids such as, for example, hydrochloric or phosphoric acids, or such organic acids as acetic, oxalic, tartaric, mandelic, and the like. Salts formed with the free carboxyl groups can also be derived from inorganic bases such as, for example, sodium, potassium, ammonium, calcium, or ferric hydroxides, and such organic bases as isopropylamine, trimethylamine, histidine, procaine and the like. [0082]
  • Upon formation, solutions will be administered in a manner compatible with the dosage formulation and in such a manner as it therapeutically effective. The formulations are easily administered in a variety of dosage forms such as injectable solutions, drug release capsules and the like. [0083]
  • In other embodiments, one may desire a topical application of compositions disclosed herein. Such compositions may be formulated in creams, lotions, solutions, or in solid form depending upon the particular application. The formulation of pharmaceutically acceptable vehicles for topical administration is well known to one of skill in the art (see, i.e., “Remington's Pharmaceuticals Sciences”, 15[0084] th Edition). Variation of the dosage of the compositions disclosed herein, will necessarily depend upon the particular subject, and the nature of the condition(s) being treated.
  • For parenteral administration in an aqueous solution, for example, the solution should be suitably buffered if necessary and the liquid diluent first rendered isotonic with sufficient saline or glucose. These particular aqueous solutions are especially suitable for intravenous, intramuscular, subcutaneous and intraperitoneal administration. In this connection, sterile aqueous media which can be employed will be known to those of skill in the art in light of the present disclosure. For example, one dosage could be dissolved in 1 ml of isotonic NaCl solution and either added to 1000 ml of hypodermic or intravenous fluid or injected at the proposed site of infusion, (see, for example, “Remington's Pharmaceutical Sciences”, 15[0085] th Edition, pages 1035-1038 and 1570-1580). Some variation in dosage will necessarily occur depending on the condition of the subject being treated. The person responsible for administration will, in any event, determine the appropriate dose for the individual subject. Moreover, for human administration, preparations should meet sterility, pyrogenicity, general safety and purity standards as required by FDA Office of Biologics standards.
  • EXAMPLE
  • Assays For Candidate Substances [0086]
  • Expression Of Human 5α-Reductases. For the preparation of rat 1A cells expressing different types of human 5α-reductases, cDNAs for the [0087] human type 1 and 2 5α-reductases were isolated from human prostate λgt11 and PC-3 cell λZAP II cDNA libraries using the published sequence of the 5α-reductases, PCR and standard library screening techniques. The type 1 and 2 cDNAs were subcloned into the retroviral expression vector pMV7 and high titer stocks of virus containing the type 1 and 2 cDNAs were generated using the packaging cells BOSC 23 293. Rat 1A cells were infected with virus and cells containing integrated retrovirus were selected for G418 resistance (Brown and Scott, 1987).
  • Assay Of 5α-Reductase. Microsomes were prepared from rat 1A cells expressing specific types of human 5α-reductase. The enzymic assay was based on the measurement of 5α-DHT production from testosterone in the presence of microsomes prepared from rat 1A cells containing either the [0088] type 1 or type 2 human 5α-reductase. The amount of labeled testosterone and dihydrotestosterone in extracts was determined by thin layer chromatography and scanning on a AMBIS radioanalytic scanner. The concentration of test compound inhibiting the conversion of testosterone to dihydrotestosterone by 50% (IC50) was determined by interpolation between appropriate data points.
  • Inhibition Of 5α-Reductase Activity. [0089]
  • Previously, we showed that two natural products, the green tea catechins, epicatechin gallate (ECG) and epigallocatechin gallate (EGCG) and unsaturated fatty acids were inhibitors of human 5α-reductase. A structure-activity relationship study was initiated to explore the structural requirements for this activity. Data for this study are summarized in Tables 1-7 and FIGS. [0090] 1-6.
  • 1. FLAVANOIDS [0091]
  • A variety of naturally occurring flavanoids with structures related to the tea catechins were tested (FIG. 1, Table 1). [0092]
    TABLE 1
    Cell Free Assay HRED1 HRED2
    Isoenzyme HRED1 % Inhibition HRED2 % Inhibition
    Compound IC50(uM) @ 100 uM IC50(uM) @ 100 uM
     1. Epicatechin 11 100 60 83
    Gallate
     2. Epigallocatechin
    Gallate 15 99 74 74
     3. Myricetin 23 96 >100 31
     4. Quercitin 23 96 >100 14
     5. Baicalein 29 79 99 51
     6. Fisetin 57 97 >100  4
     7. Biochanin a >100 50 17 74
     8. Kaempferol >100 22 12 62
     9. Flavone >100 20 >100 (−52)  
    10. Genistein >100 16 23 76
    11. Epigallocatechin >100 15 >100  3
    12. Epicatechin >100 14 >100  4
    13. Morin >100 6 >100 33
    14. Alpha-naptho- >100 6 >100 (−13) 
    flavone
    15. Taxifolin >100 5 >100  5
    16. Rutin >100 4 >100  0
    17. Daidzein >100 3 29 69
    18. Beta-naptho- >100 3 >100  4
    flavone
    19. Chrysin >100 2 >100  1
  • The tea catechins, ECG and EGCG, had the highest activity of the tested flavanoids and were better inhibitors of the type 1 (HRED1) than the type 2 (HRED2) isoenzyme of 5α-reductase. The tea catechins epicatechin (EC) and epigallocatechin (EGC) had little activity. Four flavanoids, myricetin, quercitin, baicalein and fisetin had significant (IC50<100 μM) activity and were more active against the [0093] type 1 than the type 2 isoenzyme. Biochanin A, kaempferol, genistein, and diadzein were effective inhibitors of the type 2 but not type 1 isoenzyme. Comparison of the activities of chrysin, kaempferol, morin, myricetin, and quercitin indicate the importance of B-ring hydroxyl groups, especially in a catechol or pyrogallol configuration, and perhaps the importance of the hydroxyl at position 3 for activity against the type 1 isozyme. Rutin, the 3-rutinose glycoside of quercitin was ineffective against either isoenzyme (IC50>100 μM). The inactivity of rutin compared to quercitin is either due to the presence of the oligosaccharide rutinose (perhaps due to steric hindrance) or modification of the hydroxyl at position 3. Taxifolin, a flavanone, was ineffective against either isozyme (IC50>100 μM). The weak activity of taxifolin is most likely due to the absence of the 2,3-unsaturated bond when its activity is compared to the structurally related quercitin. When tested for inhibitory activity in whole cells, most flavanoids showed little or no activity against the type 1 isoenzyme, perhaps indicating limited penetration of these polyhydroxy compounds across the cell membrane. In contrast to the results with the type 1 enzyme, four flavanoids, biochanin A, diadzein, kaempferol and genistein had significant inhibitory activity against the type 2 isoenzyme in the whole cell assay. The most active of these, biochanin A and diadzein, have only two and three free hydroxyl groups, respectively, and so may penetrate cells easier than other flavanoids.
  • 2. CATECHOLS [0094]
  • 5α-reductase inhibition studied with the flavanoids indicated the potential importance of catechol and pyrogallol moieties for high inhibitory activity. Therefore, a series of compounds with catechol groups was surveyed for activity (Table 2, FIG. 2). [0095]
    TABLE 2
    Cell Free Assay HRED1 HRED2
    Isoenzyme HRED1 % Inhibition HRED2 % Inhibition
    Compound IC50(uM) @ 100 uM IC50(uM) @ 100 uM
     1. Anthrarobin 4 99 50 97
     2. Bromopyro- 7 98 84 58
    gallol Red
     3. Gossypol 7 99 21 99
     4. Pyrogallol Red 15 97 >100 27
     5. Nordihydro- 19 99 50 80
    gaiaretic Acid
     6. Caffeic Acid 26 97 >100 36
    Phenethyl Ester
     7. Octyl Gallate 27 99 58 90
     8. Purpurogallin 30 81 >100 31
     9. Hydroxydopa- 42 69 >100 41
    mine
    10. Dodecylgallate 43 88 >100 36
    11. Pyrocatechol 48 85 100 47
    Violet
    12. Pyrogallol 70 60 >100 28
    13. Hematoxylin 83 59 >100 38
    14. HZIV-82 >100 43 >100  0
    15. Cnc >100 42 >100 (−75)  
    16. HZIV 90 >100 23 >100 13
    17. Caffeic Acid >100 13 >100  8
    18. HZIV 275 >100 10 >100  6
    19. Exculetin >100 7 >100 13
    20. Ellagic Acid >100 7 >100  9
    21. Catechol >100 5 >100  0
    22. Methylgallate >100 5 >100  3
    23. Fraxetin >100 2 >100  8
    24. Propylgallate >100 0 >100  0
  • Thirteen of the 24 compounds listed had IC50's below 100 μM. All were more active against the [0096] type 1 than type 2 isoenzyme. Six of these compounds, anthrarobin, dodecyl gallate, gossypol, octyl gallate, caffeic acid phenethyl ester and nordihydroguaiaretic acid were active in whole cell assays (Table 7, below). Anthrarobin was much more effective against the type 1 than type 2 isoenzyme; whereas, the other five inhibitors were equally effective inhibitors of both isoenzymes. The synthetic compound HZIV 82 showed little activity in the cell-free assay, but was very active in the whole cell assay with specificity for the type 1 isoenzyme.
  • 3. CURCUMIN AND RELATED COMPOUNDS [0097]
  • Curcumin was a very effective inhibitor of either the [0098] type 1 or type 2 isoenzyme (Table 3, FIG. 3).
    TABLE 3
    Cell Free Assay HRED1 HRED2
    Isoenzyme HRED1 % Inhibition HRED2 % Inhibition
    Compound IC50(uM) @ 100 uM IC50(uM) @ 100 uM
    1. Curcumin 3 95 5 87
    2. Tetrahydro- 80 56 29 73
    curcumin
    3. Demethoxy- >100 23 >100 42
    tetrahydrocurcumin
    4. 4-hydroxy-3- >100 10 >100 (−60)  
    methoxy-
    cinnamaldehyde
    5. Coniferol >100 10 100 49
    6. 4-(4-hydroxy-3- >100 3 >100  4
    methoxyphenol)-3-
    buten-2-one
    7. Ferulic Acid >100 0 >100 18
    8. Capsaicin >100 0 >100  8
    9. Eugenol >100 0 100 50
  • Commercially available curcumin was chemically reduced with Pt/H2 and the products, tetrahydrocurcumin and demethoxytetrahydrocurcumin, had much less activity than curcumin. However, tetrahydrocurcumin (HZIV 81-2), which is colorless compared to the bright yellow curcumin, had significant activity in the whole cell assay. The structurally related compounds 4-(4-hydroxy-3-methoxyphenol)-3-buten-2-one, ferulic acid, capsaicin, eugenol and coniferyl alcohol had little inhibitor activity (IC50>100 μM) against either isoenzyme highlighting the importance of the diferulolyl structure for activity against 5α-reductase. Nordihydroguaiaretic acid (NDGA) was also an effective inhibitor of the type 1 (IC50=19 μM) and type 2 (IC50=50 μM) isozymes in cell-free and whole cell assays, but less so than curcumin. [0099]
  • 4. QUINONES [0100]
  • A variety of quinones were tested for activity against 5α-reductase (Table 4, FIG. 4). [0101]
    TABLE 4
    Cell Free HRED1 HRED2
    Assay Isoenzyme HRED1 % Inhibition HRED2 % Inhibition
    Compound IC50(μM) @ 100 μM IC50(μM) @ 100 μM
     1. Purpurin 2 95 >100 20
     2. Alizarin 3 95 100 54
     3. Anthrarobin 4 99 50 97
     4. Menadione 6 77 5 81
     5. Coenzyme q 12 77 22 81
     6. 2,5-dichloroindophenol 15 78 17 97
     7. Alizarin Red 5 30 91 >100 8
     8. Anthrarufin 40 67 >100 13
     9. Anthrarufin 40 67 >100 13
    10. Lapachol >100 30 >100 9
    11. Anthraflavic Acid >100 27 >100 22
    12. Quinizarin >100 26 >100 7
    13. T-butylhydroxyquinone >100 19 >100 4
    14. Anthraquinone >100 6 >100 9
  • The naturally occurring anthraquinone, alizarin, was a very effective inhibitor of the [0102] type 1 but not type 2 isozymes. Alizarin Red S, which is a water soluble sulfate derivative of alizarin had little activity (IC50s>100 μM) against either isoenzyme. The charged sulfate group may prevent interaction with membrane bound 5α-reductase. Purpurin, which has an additional hydroxyl compared to alizarin, had inhibitory activity similar to alizarin. Anthraflavic acid, anthrarufin and quinizarin, which are structural isomers of alizarin without adjacent hydroxyl groups, had much less activity, emphasizing the importance of the catechol moiety for potent inhibitory activity of this class of anthroquinones. Anthraquinone was not an effective inhibitor (IC50>100 μM). Menadione, coenzyme Q, and 2,6-dichloroindophenol were potent cell-free inhibitors of boh isoenzymes. The compounds participate in quinone reductase reactions and may deplete NADPH causing the observed inhibition. In the whole cell assay, alizarin was a very effective inhibitor of the type 1 isoenzyme and menadione had moderate activity.
  • 5. EPIGALLOCATECHIN DERIVATIVES [0103]
  • The high inhibitory activity of EGCG in a cell-free assay but low in the whole cell assay led us to design and synthesize a series of derivatives of EGC to enhance activity in the whole cell assay (Table 5, FIG. 5). [0104]
    TABLE 5
    Cell Free HRED1 HRED2
    Assay Isoenzyme HRED1 % Inhibition HRED2 % Inhibition
    Compound IC50(μM) @ 100 μM IC50(μM) @ 100 μM
     1. EGCG 12 99 73 76
     2. HZIV 160 29 99 76 96
     3. HZIV 134 20 99 67 94
     4. HZIV 92 23 98 >100 45
     5. HZIV 120 23 99 66 97
     6. HZIV 142 25 97 63 93
     7. HZIV 68 29 93 99 51
     8. HZIV 75 29 97 >100 21
     9. HZIV 166 30 98 78 74
    10. HZIV 63 311 94 >100 20
    11. HZIV 169 47 90 >100 39
    12. HZIV 74 48 85 >100 24
    13. HZIV 144 49 88 >100 38
    14. HZIV 168 49 98 73 92
    15. HZIV 166 59 95 71 84
    16. BGC 62 61 >100 30
    17. HZIV 107 98 52 >100 39
    18. HZIV 145 >100 35 >100 8
    19. HZIV 148 >100 31 >100 0
    20. HZIV 109 >100 17 >100 0
  • The studies showed that derivatization of the hydroxyl groups of EGCG with methyl or acetate groups leads to the loss of inhibitory activity in the cell-free assay and no enhancement of the lower whole cell assay inhibitory activity. We, therefore, limited structure-activity studies to changes in the gallate ester moiety of EGCG to enhance inhibitory activity in the whole cell assay. Twenty of these structural changes are summarized in Table 5. The most significant structural change lading to activity in the whole cell assay was introduction of fatty acid ester in place of the gallic acid group of EGCG. In particular, fatty acids with some degree of unsaturation had good inhibitory activity against both isoenzymes of 5α-reductase in the whole cell assay. The most potent of these derivatives was one with γ-linolenic acid esterified to the 3-hydroxyl of EGC. Certain fatty acids with a single unsaturated bond were also active. For example, [0105] HZIV 160, the myristoleic ester of BGC was effective in both assay systems. Fatty acids with less unsaturation are less susceptible to oxidation and so may be more suitable modifying agents.
    TABLE 6
    Cell Free HRED1 HRED2
    Assay Isoenzyme HRED1 % Inhibition HRED2 % Inhibition
    Compound IC50(μM) @ 100 μM IC50(μM) @ 100 μM
    1. Gamma-Linolenic Acid 5 99 11 89
    C18:3 CIS 6.9,12
    2. Crocetin 7 70@30 >100 20@30
    3. Alpha-Linolenic Acid 8 99 9 84
    C18:3 CIS 9,12,15
    4. Linoleic Acid 9 99 19 85
    C18:2 CIS 9,12
    5. Oleic Acid C18:1 CIS 10 99 42 86
    9
    6. Conjugated 10 99 30 81
    Octadecadienonic Acid
    7. 5,8,11,14-Eocpsatetraynoic Acid 15 97 3 81
    8. Stearic Acid C18:0 27 71 >100 35
  • [0106]
    TABLE 7
    Whole Cell HRED1 HRED2
    Assay Isoenzyme HCRED1 % Inhibition HCRED2 % Inhibition
    Compound IC50(μM) @ 100 μM IC50(μM) @ 100 μM
    1. HZIV 82 3 79 >100 15
    2. Dodecylgallate 3 99 7 98
    3. Anthrarobin 6 91 >100 31
    4. Alizarin 6 75 >100 27
    5. Gossypol 7 100 6 99
    6. HZIV 160 7 99 8 98
    7. Octyl Gallate 7 99 18 94
    8. Caffeic Acid Phenethyl 8 99 7 98
    Ester
    9. HZIV 142 8 99 14 98
    10. Curcumin 9 99 7 99
    11. Nordihydroguaiaretic 19 99 22 99
    Acid
    12. HZIV 165 28 97 32 98
    13. HZIV 168 28 93 41 94
    14. HZIV 81-2 36 81 7 92
    15. HZIV 148 42 90 74 81
    16. HZIV 75 43 83 62 72
    17. HZIV 120 49 97 57 96
    18. Menadione 51 82 79 62
    19. HZIV 166 58 89 72 83
    20. Biochanin A 64 64 5 93
    21. HZIV 92 64 94 80 62
    22. Kaempferol 79 60 20 85
    23. Daidzein 10 13 7 89
    24. Baicalein >100 24 >100 4
    25. Fisetin >100 42 >100 27
    26. EGCG >100 11 >100 5
    27. Myricetin >100 11 >100 11
    28. Purpurin >100 47 >100 7
    29. Quercetin >100 15 >100 29
    30. Alizarin Red S >100 28 >100 1
    31. Genistein >100 22 20 89
    32. HZIV 123 >100 48 >100 8
    33. HZIV 107 >100 23 >100 2
    34. Catechol >100 9 >100 3
    35. Daidzein >100 9 58 87
    36. Pyrogallol >100 7 >100 15
    37. EC >100 0 >100 1
    38. EGC >100 15 >100 1
    39. ECG >100 0 >100 0
    40. EGCG >100 6 >100 0
    41. HZIV 90 >100 34 >100 14
    42. HZIV 63 >100 12 >100 7
    43. HZIV 68 >100 40 >100 34
    44. HZIV 144 >100 12 >100 7
    45. HZIV 81-3 >100 28 19 80
    46. HZIV 145 >100 8 >100 9
    47. Methyl Gallate >100 0 >100 0
    48. Propyl Gallate >100 5 >100 0
    49. Isopropyl Gallate >100 0 >100 0
    50. Gallic Acid >100 13 >100 0
    51. Pyrogallol >100 5 >100 6
    52. HZIV 169 >100 10 >100 0
    53. Gamma-Linolenic 22 91 20 86
    54. Etya 22 67 2 86
    55. Alpha-Linolenic 29 82 23 86
    56. Linoleic 40 78 25 77
    57. Oleic Acid 83 58 >100 45
    58. Stearic Acid >100 10 >100 23
    59. Alpha-Linolenic Acid >100 24 >100 18
    Me Ester
    60. Gamma-Linolenic Acid >100 12 >100 11
    Chol Ester
    61. Gamma-Linolenic Acid >100 49 >100 26
    Me Ester
  • As found previously in our lab, the greater the degree of unsaturation, the better the inhibitory activity of the fatty acid. Since unsaturated fatty acids are easily prone to oxidation which may comprise their usefulness, we examined some unsaturated fatty acids less prone to oxidation. The synthetic fatty acids, conjugated octadecadienoic acid (CODA) (cis or trans-9,11 or 10,12 octadecadienoic acid) and 5,8,11,14-eicosatetraynoic acid (ETYA), were good inhibitors of both isoenzymes. CODA and ETYA had IC50s of 10 and 15 (type 1) and 30 and 3 (type 2) μM, respectively. The naturally occurring fatty acid, γ-linolenic acid, has IC50 of 3 μM for both isoenzymes. Fatty acids such as ETYA may be useful for derivatizing other 5α-reductase inhibitors to enhance cellular uptake and promote in vivo activity of 5α-reductase inhibitors. Methyl and cholesterol esters of γ-linolenic acid had little activity in the whole cell assay (TABLE 7) and so the activity of EGC esterified to γ-linolenic acid is unlikely due to intracellular hydrolysis of these esters. [0107]
  • Active 5α-reductase inhibitors shown in Tables 1-7 are polyphenols or their derivatives and are easily oxidized or hydrolyzed within several hours to several days, especially in the presence of air or oxygen and at a pH above 7.0. We have found that these compounds are more stable to oxidation or hydrolysis by maintaining the pH of the solutions of these compounds at a pH below 7.0. More than 80% of the oxidation of hydrolysis can be prevented by the addition of an inorganic acid, such as hydrochloric acid, sulfuric acid, or phosphoric acid, or an organic acid, such as citric acid or acetic acid. [0108]
  • The references listed below and cited in the disclosure are: [0109]
  • 1. Anderson and Liao. Nature, 219: 277-279, 168. [0110]
  • 2. Brown and Scott, DNA Cloning, A Practical Approach, Vol. III; 189-212, 1987. [0111]
  • 3. Bruchovsky and Wilson. J. Biol. Chem 243: 5953-5960, 1968. [0112]
  • 4. Chakrabarry et al., J. Invest. Dermatol, 74: 5-8, 1980. [0113]
  • 5. Diani et al., J. Clin. Endocrinol. Metab. 74: 345-350, 1992. [0114]
  • 6. Frost and Gomez, Adv. Biol. Skin, 12:403-442, 1972. [0115]
  • 7. Frost et al., J. Invest. Dermatol, 61:159-167, 1973. [0116]
  • 8. Hamilton and Montagna, Amer. J. Anat., 86:191-233, 1950. [0117]
  • 9. Hiipakka et al., J. Steroid Biochem. Molec. Biol., 45: 539-548. [0118]
  • 10. Hirsch et al., Proc. Natl. Acad. Sci. USA, 90: 5277-5281, 1993. [0119]
  • 11. Liang and Liao, Biochem. J.285: 557-562, 1992. [0120]
  • 12. Liang and Liao, J. Invest. Dermatol. 109: 152-157, 1997. [0121]
  • 13. Liang et al., Endocrinology 112: 1460-1468, 1983. [0122]
  • 14. Liao and Hiipakka, Biophys. Biochem. Res. Commun. 214: 833-838, 1995. [0123]
  • 15. Liao et al., J. Steroid Biochem, 34: 41-51, 1989. [0124]
  • 16. Liao et al., Cancer Letters, 96: 239-243, 1995. [0125]
  • 17. Luderschmidt et al., J. Invest. Dermatol., 83: 157-160, 1984. [0126]
  • 18. Randall, Clin. Endrocrinol 40: 439-457, 1994. [0127]
  • 19. Russell and Wilson, Ann. Rev. Biochem. 63: 25-61, 1994. [0128]
  • 20. Stoner et al., J. Urol. 147: 1298-1302, 1992. [0129]
  • 21. Takayasu et al., Endocrinology 90: 73-79, 1972. [0130]
  • 22. Voight and Hsia, Endocrinology, 92: 1216-1222, 1973. [0131]
  • 23. Weissmann et al., J. Invest. Dermatol., 82: 522-525, 1985. [0132]
  • 24. Williams, Clin. Pharmacokinetics, 10: 392-403, 1985. [0133]
  • 25. Wuest and Lucky, Skin Pharmacol., 2: 103-113, 1989. [0134]

Claims (11)

What is claimed is:
1. A process of inhibiting 5α-reductase activity comprising the step of exposing 5α-reductase to an effective inhibiting amount of a compound of any of Tables 1-7.
2. A process of treating prostate cancer in a subject in need of such treatment comprising the step of administering to the subject an effective therapeutic amount of a compound of any of Tables 1-7.
3. A process of treating breast cancer in a subject in need of such treatment comprising the step of administering to the subject an effective therapeutic amount of a compound of any of Tables 1-7.
4. A process of treating baldness in a subject in need of such treatment comprising the step of administering to the subject an effective therapeutic amount of a compound of any of Tables 1-7.
5. A process of treating a skin disorder in a subject in need of such treatment comprising the step of administering to the subject an effective therapeutic amount of a compound of any of Tables 1-7.
6. A process of treating obesity in a subject in need of such treatment comprising the step of administering to the subject an effective therapeutic amount of a compound of any of Tables 1-7.
7. A process of treating benign prostatic hyperplasia or prostatitis in a subject in need of such treatment comprising the step of administering to the subject an effective therapeutic amount of a compound of any of Tables 1-7.
8. A compound of the formula:
Figure US20030144346A1-20030731-C00004
where x is —NHCH2CH2— or —CH═CH—;
R1, R2 and R3 each may be —H, —OH or —OCH3, provided that only one of R1, R2, and R3 may be —H;
R4, R5 and R6 each may be —H, —OH, —OCH3 or —N(CH3)2, provided that only one of R4, R5 and R6 may be —H; and
n is 0 or 1.
9. A compound of the formula:
Figure US20030144346A1-20030731-C00005
where R is a chain with 2 to 20 atoms from the group consisting of carbon, oxygen, sulfur, and nitrogen, without or with one to four double bonds and additional hydrogen.
10. A compound of the formula:
Figure US20030144346A1-20030731-C00006
where R is
Figure US20030144346A1-20030731-C00007
11. A method for the stabilization of compounds shown in Tables 1-7 comprising adding an effective ambient of an inorganic acid, an organic acid, or a natural product that contains these acids to maintain the acidity of the therapeutic preparations of these compounds at pH 3.0 to pH 6.8.
US10/294,331 1997-10-31 2002-11-14 Methods and compositions for regulation of 5-alpha reductase activity Abandoned US20030144346A1 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
US10/294,331 US20030144346A1 (en) 1997-10-31 2002-11-14 Methods and compositions for regulation of 5-alpha reductase activity

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US6377097P 1997-10-31 1997-10-31
US09/530,443 US6576660B1 (en) 1997-10-31 1998-10-30 Methods and compositions for regulation of 5-α-reductase activity
US10/294,331 US20030144346A1 (en) 1997-10-31 2002-11-14 Methods and compositions for regulation of 5-alpha reductase activity

Related Parent Applications (2)

Application Number Title Priority Date Filing Date
PCT/US1998/023041 Continuation WO1999022728A1 (en) 1997-10-31 1998-10-30 Methods and compositions for regulation of 5-alpha reductase activity
US09/530,443 Continuation US6576660B1 (en) 1997-10-31 1998-10-30 Methods and compositions for regulation of 5-α-reductase activity

Publications (1)

Publication Number Publication Date
US20030144346A1 true US20030144346A1 (en) 2003-07-31

Family

ID=22051389

Family Applications (2)

Application Number Title Priority Date Filing Date
US09/530,443 Expired - Lifetime US6576660B1 (en) 1997-10-31 1998-10-30 Methods and compositions for regulation of 5-α-reductase activity
US10/294,331 Abandoned US20030144346A1 (en) 1997-10-31 2002-11-14 Methods and compositions for regulation of 5-alpha reductase activity

Family Applications Before (1)

Application Number Title Priority Date Filing Date
US09/530,443 Expired - Lifetime US6576660B1 (en) 1997-10-31 1998-10-30 Methods and compositions for regulation of 5-α-reductase activity

Country Status (6)

Country Link
US (2) US6576660B1 (en)
EP (1) EP1027045A4 (en)
JP (1) JP4738592B2 (en)
KR (1) KR20010031501A (en)
AU (1) AU1289899A (en)
WO (1) WO1999022728A1 (en)

Cited By (14)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20050171084A1 (en) * 2002-03-27 2005-08-04 Cairns William J. Methods of treatment with lxr modulators
US20050175565A1 (en) * 2002-06-21 2005-08-11 L'oreal Use of taurine for the treatment of alopecia
US20060159714A1 (en) * 2003-07-07 2006-07-20 Jean-Noel Thorel Use of an antioxidant in a dermatological and/or cosmetic composition
US20100034904A1 (en) * 2007-03-21 2010-02-11 Giuliani S.P.A. Composition comprisong rutin and polyunsaturated fatty acid having an inhibitory activity on 5-alph reductase
US8147825B2 (en) 2004-01-22 2012-04-03 University Of Miami Topical co-enzyme Q10 formulations and methods of use
US8454945B2 (en) 2007-03-22 2013-06-04 Berg Pharma Llc Topical formulations having enhanced bioavailability
US8557864B2 (en) 2007-09-19 2013-10-15 Nagoya Industrial Science Research Institute Agent having neurotrophic factor-like activity
US9896731B2 (en) 2009-05-11 2018-02-20 Berg Llc Methods for treatment of oncological disorders using an epimetabolic shifter (coenzyme Q10)
US9901542B2 (en) 2013-09-04 2018-02-27 Berg Llc Methods of treatment of cancer by continuous infusion of coenzyme Q10
US10376477B2 (en) 2011-04-04 2019-08-13 Berg Llc Method of treating or preventing tumors of the central nervous system
US10668028B2 (en) 2008-04-11 2020-06-02 Berg Llc Methods and use of inducing apoptosis in cancer cells
US10933032B2 (en) 2013-04-08 2021-03-02 Berg Llc Methods for the treatment of cancer using coenzyme Q10 combination therapies
US10973763B2 (en) 2011-06-17 2021-04-13 Berg Llc Inhalable pharmaceutical compositions
US11400058B2 (en) 2010-03-12 2022-08-02 Berg Llc Intravenous formulations of coenzyme Q10 (CoQ10) and methods of use thereof

Families Citing this family (129)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5385936A (en) 1990-07-12 1995-01-31 The United States Of America As Represented By The Secretary Of The Department Of The Health And Human Services Gossypol acetic acid for the treatment of cancer
US6908910B2 (en) 1993-08-06 2005-06-21 The Children's Medical Center Corporation Estrogenic compounds as anti-mitotic agents
US5504074A (en) 1993-08-06 1996-04-02 Children's Medical Center Corporation Estrogenic compounds as anti-angiogenic agents
JP4738592B2 (en) 1997-10-31 2011-08-03 アーチ・デヴェロップメント・コーポレイション Methods and compositions for modulating 5α-reductase activity
US6696484B2 (en) 1997-10-31 2004-02-24 University Of Chicago Office Of Technology And Intellectual Property Method and compositions for regulation of 5-alpha reductase activity
US5958946A (en) * 1998-01-20 1999-09-28 Styczynski; Peter Modulation of hair growth
IL142650A (en) * 1998-04-08 2007-06-03 Galmed Int Ltd Use of bile acid or bile salt fatty acids conjugates for the preparation of pharmaceutical compositions for reducing cholesterol, treating fatty liver and treating hyperglycemia and diabetes
GB0000710D0 (en) * 1999-02-06 2000-03-08 Zeneca Ltd Drug combination
US7087592B1 (en) 1999-08-23 2006-08-08 Entre Med, Inc. Compositions comprising purified 2-methoxyestradiol and methods of producing same
GB9920548D0 (en) * 1999-08-31 1999-11-03 Rhone Poulenc Rorer Sa Treatment of hepatocellular carcinoma
EP1232756A4 (en) * 1999-10-29 2006-03-22 Takara Bio Inc Drugs, drinks or foods
FI20000004A0 (en) * 2000-01-03 2000-01-03 Slk Foundation Flavonoidilääke ...
FR2803747B1 (en) * 2000-01-18 2003-12-26 Pharmascience Lab USE OF ISOFLAVONES AND / OR EXTRACTS OF AFRICAN PLUM IN PHARMACY, COSMETICS AND AS A FOOD ADDITIVE.
FR2803749B1 (en) * 2000-01-18 2006-09-22 Pharmascience Lab USE OF AT LEAST ONE FATTY ESTER FOR THE PREPARATION OF A COMPOSITION INTENDED TO INHIBIT THE ACTIVITY OF ALPHA-REDUCTASE, IN PHARMACY IN PARTICULAR DERMATOLOGY, COSMETICS AND AS A FOOD ADDITIVE
US6610749B2 (en) 2000-02-18 2003-08-26 Arch Development Corporation Polyhydroxylated benzene-containing compounds
IL146010A0 (en) * 2000-02-18 2002-07-25 Arch Dev Corp Polyhydroxylated benzene-containing compounds
US6518261B2 (en) * 2000-03-17 2003-02-11 Oncology Sciences Corporation Use of eugenol in combination with other chemopreventative agents as prophylaxis for cancers
WO2001074345A2 (en) * 2000-03-31 2001-10-11 Jonathan Ingram Isoflavones for treatment of obesity
FR2808192A1 (en) * 2000-04-28 2001-11-02 Oreal EPICHATCHIN AS INHIBITOR OF NO-SYNTHASE AND USES
US7084126B1 (en) * 2000-05-01 2006-08-01 Healthpartners Research Foundation Methods and compositions for enhancing cellular function through protection of tissue components
DE10029770A1 (en) * 2000-06-16 2001-12-20 Transmit Technologietransfer Treatment of solid tumors, especially brain tumors, using curcumin or derivative to inhibit peritumoral edema and tumor growth and kill the tumor cells
US7427689B2 (en) * 2000-07-28 2008-09-23 Georgetown University ErbB-2 selective small molecule kinase inhibitors
US7135581B2 (en) 2000-08-18 2006-11-14 Entremed, Inc. Antiangiogenic agents
FR2817749A1 (en) * 2000-12-13 2002-06-14 Aventis Pharma Sa NEW USE OF CHALCON CLASS COMPOUNDS
US6555573B2 (en) 2000-12-21 2003-04-29 The Quigley Corporation Method and composition for the topical treatment of diabetic neuropathy
WO2002062348A1 (en) * 2001-02-05 2002-08-15 Oncology Sciences Corporation Eugenol, optionally in combination with 2-methoxyestradiol, as a cancer chemopreventative agent
ITVR20010031A1 (en) * 2001-03-12 2002-09-12 Hisanori Suzuki USE OF EPIGALLOCATECHIN-3-GALLATO OR ITS DERIVATIVES IN THE PROPHYLAXIS AND TREATMENT OF NEURODEGENERATIVE DISEASES.
EP1383492A4 (en) * 2001-03-23 2008-12-24 Napro Biotherapeutics Inc Molecular conjugates for use in treatment of cancer
PL205635B1 (en) * 2001-04-09 2010-05-31 Inst Farmaceutyczny Novel derivatives of genistein and pharmacological agents containing such derivatives
US8975246B2 (en) 2001-04-17 2015-03-10 Galmed Research And Development Ltd. Bile acid or bile salt fatty acid conjugates
US7432304B2 (en) * 2001-05-30 2008-10-07 The Regents Of The University Of Michigan Small molecule antagonists of Bcl-2 family proteins
US7595378B2 (en) 2001-06-13 2009-09-29 Genmab A/S Human monoclonal antibodies to epidermal growth factor receptor (EGFR)
JP4179494B2 (en) * 2001-10-23 2008-11-12 株式会社カネカ Peroxisome proliferator-responsive receptor ligand agent
AU2002360335A1 (en) * 2001-11-01 2003-05-12 The Regents Of The University Of Michigan Small molecule inhibitors targeted at bcl-2
PT1448186E (en) 2001-11-19 2009-03-24 Medigene Ag Medicament for the treatment of viral skin and tumour diseases
US20030236439A1 (en) * 2002-01-30 2003-12-25 Gregory Agoston Non-steroidal analogs of 2-methoxyestradiol
US20050118232A1 (en) * 2002-02-12 2005-06-02 Elvira Pistolesi N-acyl-phosphatidyl-ethanolamines and/or mixtures of n-acyl-ethanolamines with phosphatidic acids or lysophosphatidic acids
US7972632B2 (en) 2003-02-28 2011-07-05 Unigen Pharmaceuticals, Inc. Identification of Free-B-Ring flavonoids as potent COX-2 inhibitors
US7108868B2 (en) 2002-03-22 2006-09-19 Unigen Pharmaceuticals, Inc. Isolation of a dual cox-2 and 5-lipoxygenase inhibitor from acacia
FR2847805B1 (en) * 2002-11-28 2005-01-07 Oreal METHOD FOR THE DENSIFICATION OF HUMAN KERATIN FIBERS USING A PARTICULAR CATALYTIC SYSTEM
EP1348421A3 (en) * 2002-03-25 2004-01-02 L'oreal A method of thickening and styling human keratinous fibres using a particular catalytic system
US6790979B2 (en) * 2002-04-17 2004-09-14 University Of North Carolina At Chapel Hill Curcumin analogues and uses thereof
US7355081B2 (en) 2002-04-17 2008-04-08 The University Of North Carolina At Chapel Hill Curcumin analogues and uses thereof
US8034387B2 (en) 2002-04-30 2011-10-11 Unigen, Inc. Formulation of a mixture of free-B-ring flavonoids and flavans for use in the prevention and treatment of cognitive decline and age-related memory impairments
EP2108370A1 (en) 2002-04-30 2009-10-14 Unigen Pharmaceuticals, Inc. Formulation of a mixture of free-B-ring flavonoids and flavans as a therapeutic agent
US7790706B2 (en) 2002-06-14 2010-09-07 The University Of Edinburgh Treatment of inflammation with 5α reduced metabolites
GB0213745D0 (en) * 2002-06-14 2002-07-24 Univ Edinburgh Enzyme
AU2007229399B2 (en) * 2002-06-21 2010-12-23 L'oreal Use of taurine or derivatives thereof for the treatment of alopecia
AU2003266313B2 (en) * 2002-08-27 2009-10-22 Societe Des Produits Nestle S.A. Preventing or treating epithelial tissue damage or hair loss
US7083813B2 (en) * 2002-11-06 2006-08-01 The Quigley Corporation Methods for the treatment of peripheral neural and vascular ailments
US20040097429A1 (en) * 2002-11-18 2004-05-20 Nieuwenhuizen Arie Gijsbert Method for the reduction of the mammalian appetite
US7122573B2 (en) * 2002-12-06 2006-10-17 Sri International Analogs of green tea polyphenols as chemotherapeutic and chemopreventive agents
AU2003298241A1 (en) * 2003-02-19 2004-09-09 Unilever Plc Topical composition containing alkyl gallate or alkyl hydroxybenzoate or phenol alkyloxy as agent against acne
WO2004075844A2 (en) * 2003-02-26 2004-09-10 Unigen Pharmaceuticals, Inc. Formulation for use in the prevention and treatment of carbohydrate induced diseases and conditions
US7208486B2 (en) 2003-03-07 2007-04-24 Schering Corporation Substituted azetidinone compounds, processes for preparing the same, formulations and uses thereof
ES2311806T3 (en) 2003-03-07 2009-02-16 Schering Corporation AZETIDINONA COMPOSITE SUBSTITUTED, FORNULATIONS AND USES OF THE SAME FOR THE TREATMENT OF HYPERCHOLESTEROLEMIA.
US7326734B2 (en) * 2003-04-01 2008-02-05 The Regents Of The University Of California Treatment of bladder and urinary tract cancers
BRPI0409179A (en) 2003-04-04 2006-05-02 Unigen Pharmaceuticals Inc formulation of dual cyclooxygenase (cox) and lipoxygenase (lox) inhibitors for mammalian skin care
CN103585136A (en) * 2003-05-20 2014-02-19 约翰霍普金斯大学 Methods and compositions for delivery of catecholic butanes for treatment of tumors
FR2856304B1 (en) * 2003-06-20 2006-03-03 Natural Product Consulting COMPOSITION FOR THE PREVENTION OF INFECTIONS OF THE URINARY SYSTEM
US20060025337A1 (en) * 2003-07-01 2006-02-02 President And Fellows Of Harvard College Sirtuin related therapeutics and diagnostics for neurodegenerative diseases
AU2004253579B2 (en) * 2003-07-01 2010-12-23 Biomol International L.P. Sirt1 modulators for manipulating cells/organism lifespan/stress response
US20050008712A1 (en) * 2003-07-08 2005-01-13 Debasis Bagchi Compositions incorporating high-caffeine green tea extract and related methods for promoting healthy body weight
FR2859104A1 (en) * 2003-08-29 2005-03-04 Saint Laurent Parfums MOLECULES WITH ANTI-ADIPOGENIC ACTIVITY ON HUMAN ADIPOCYTE CELLS
WO2005027901A1 (en) * 2003-09-25 2005-03-31 Tel Aviv University Future Technology Development L.P. Compositions and methods using same for treating amyloid-associated diseases
US20050079235A1 (en) * 2003-10-09 2005-04-14 Eggert Stockfleth Use of a polyphenol for the treatment of actinic keratosis
EP2292226A3 (en) * 2003-10-09 2011-08-10 MediGene AG The use of a polyphenol for the treatment of a cancerous or pre-cancerous lesion of the skin
KR20050037102A (en) * 2003-10-17 2005-04-21 한국 한의학 연구원 Compositions for inhibiting obesity
WO2005046584A2 (en) * 2003-11-05 2005-05-26 Osteoscreen, Inc. Stimulation of hair growth by ginkgo biloba flavanoids
JP2007527418A (en) * 2003-12-29 2007-09-27 プレジデント・アンド・フェロウズ・オブ・ハーバード・カレッジ Composition for treating or preventing obesity and insulin resistance disorders
US8017634B2 (en) 2003-12-29 2011-09-13 President And Fellows Of Harvard College Compositions for treating obesity and insulin resistance disorders
EP1566439A1 (en) * 2004-02-20 2005-08-24 Nestec S.A. Preventing or treating obesity
AU2005222934B2 (en) 2004-03-12 2010-06-17 Entremed, Inc. Antiangiogenic agents
JP2007530568A (en) * 2004-03-25 2007-11-01 ザ リージェンツ オブ ザ ユニバーシティ オブ ミシガン Gossypol co-crystal and use thereof
JP4904498B2 (en) * 2004-03-29 2012-03-28 財団法人ヒューマンサイエンス振興財団 Natural compounds that cause RAR activation
EP1734963A4 (en) * 2004-04-02 2008-06-18 Merck & Co Inc Method of treating men with metabolic and anthropometric disorders
WO2005113069A2 (en) * 2004-05-14 2005-12-01 Research Development Foundation Use of circumin and analogues as inhibitors of acc2
JP3790767B2 (en) * 2004-06-30 2006-06-28 森下仁丹株式会社 Fat metabolism improving composition
WO2006004722A2 (en) * 2004-06-30 2006-01-12 Biomol Research Laboratories, Inc. Compositions and methods for selectively activating human sirtuins
US20060009430A1 (en) * 2004-07-01 2006-01-12 Kelly Gregory J Composition for the prevention and treatment of the detrimental effects of dihydrotestosterone
US7618615B2 (en) 2004-08-13 2009-11-17 Healthpartners Research Foundation Methods for providing neuroprotection for the animal central nervous system against neurodegeneration caused by ischemia
US9216161B2 (en) 2004-08-13 2015-12-22 Healthpartners Research Foundation Methods of treating Huntington's disease comprising administering metal chelators to the upper one-third of the nasal cavity
US7544816B2 (en) 2004-08-19 2009-06-09 The Hong Kong Polytechnic University (−)-Epigallocatechin gallate derivatives for inhibiting proteasome
CN101072764B (en) * 2004-08-19 2013-07-10 香港理工大学 (-)-epigallocatechin gallate derivatives for inhibiting proteasome
CA2595486A1 (en) * 2005-01-20 2006-07-27 Sirtris Pharmaceuticals, Inc. Use of sirtuin-activating compounds for treating flushing and drug induced weight gain
EP1849779A4 (en) * 2005-01-26 2010-07-14 Suntory Holdings Ltd Esterified catechin, process for producing the same, food and drink or cosmetic containing the same
EP1841779A2 (en) * 2005-01-28 2007-10-10 Barry Callebaut AG Use of cacao polyphenols for treating a prostate hyperplasia, a specific cacao extract and applications
WO2006104762A2 (en) 2005-03-25 2006-10-05 Merck & Co., Inc. Method of treating men with testosterone supplement and 5alpha-reductase inhibitor
PT1871353E (en) * 2005-04-15 2011-03-29 Einstein Coll Med Vitamin k for prevention and treatment of skin rash secondary to anti-egfr therapy
WO2006138418A2 (en) * 2005-06-14 2006-12-28 President And Fellows Of Harvard College Improvement of cognitive performance with sirtuin activators
US20070149466A1 (en) * 2005-07-07 2007-06-28 Michael Milburn Methods and related compositions for treating or preventing obesity, insulin resistance disorders, and mitochondrial-associated disorders
KR101273027B1 (en) * 2005-08-20 2013-06-10 (주)아모레퍼시픽 Composition for inhibiting sebum secretion and anti-obesity comprising kaempferol
FR2893026B1 (en) * 2005-11-08 2010-09-17 Polaris NOVEL POLYPHENOLIC LIPOSOLUBLE DERIVATIVES AND THEIR USE AS ANTIOXIDANTS
FR2892923B1 (en) 2005-11-08 2009-01-16 Engelhard Lyon Sa USE OF PARA-COUMARIC OR PARA-HYDROXYCINNAMIC ACID DERIVATIVES IN COSMETIC OR DERMATOLOGICAL COMPOSITIONS.
US7700136B2 (en) * 2005-11-14 2010-04-20 Bionovo, Inc. Scutellaria barbata extract for the treatment of cancer
US7537774B2 (en) * 2005-12-23 2009-05-26 Orion Therapeautics, Llc Therapeutic formulation
JP2007197387A (en) * 2006-01-27 2007-08-09 Maruzen Pharmaceut Co Ltd TESTOSTERONE 5alpha-REDUCTASE INHIBITOR
CA2646065C (en) 2006-03-20 2014-01-14 Entremed, Inc. Disease modifying anti-arthritic activity of 2-methoxyestradiol
CA2656613A1 (en) * 2006-07-03 2008-01-10 Genmab A/S Prevention of rash in patients undergoing anti-egfr therapy
WO2008065527A2 (en) * 2006-11-27 2008-06-05 Carlo Ghisalberti Gallic acid esters of fragrant alcohols
EP1950210B1 (en) * 2007-01-23 2013-01-16 Polaris SAS Use of fat-soluble polyphenolic derivatives as antioxidants
US9707274B2 (en) 2007-06-08 2017-07-18 Healthpartners Research & Education Methods for preventing and treating post-traumatic stress disorder (PTSD)
US8143014B2 (en) * 2007-10-02 2012-03-27 Mayo Foundation For Medical Education And Research CD38 and obesity
DE102008012988A1 (en) 2008-03-07 2009-09-10 S.W. Patentverwertungs Ltd. Composition and uses for influencing hair growth
US8815953B2 (en) * 2008-03-13 2014-08-26 Spectrum Pharmaceuticals, Inc. Formulations of vitamin K analogs for topical use
WO2010028187A2 (en) * 2008-09-03 2010-03-11 Bionovo, Inc. Methods and compositions for the treatment of cancer
US8313782B2 (en) 2008-12-18 2012-11-20 Guthery B Eugene Acne vulgaris treatment regimen
EP2424555A4 (en) * 2009-04-28 2013-02-20 Bionovo Inc Method of reducing fat accumulation and inducing weight loss
WO2010134595A1 (en) * 2009-05-21 2010-11-25 サントリーホールディングス株式会社 Anti-obesity agent comprising compound containing benzotropolone ring
US8163796B1 (en) * 2009-07-28 2012-04-24 BioChemical Solutions, LLC Treatment of cancer by oxidation-reduction potentiation of cancerostatic dicarbonyls
CN102526004A (en) * 2010-01-25 2012-07-04 四川省中医药科学院 New use of tetrahydrocurcumin
KR101247239B1 (en) * 2010-11-25 2013-03-26 아쿠아그린텍(주) Compositon for Prevention of Hair Loss or Stimulation of Hair Growth Containing Ecklonia cava Extracts Having Diekol or Diekol derived from Ecklonia cava
WO2013055913A2 (en) 2011-10-14 2013-04-18 Emory University Pgam1 inhibitors and methods related thereto
KR101361147B1 (en) * 2012-06-27 2014-02-13 한국식품연구원 Compositions for Activating hTRPA1 Comprising Extract of Compositae Plants and Uses thereof
WO2013073801A1 (en) * 2011-11-18 2013-05-23 한국식품연구원 Htrpa1-activating composition and use thereof
US20150073043A1 (en) * 2012-01-19 2015-03-12 Institut Pasteur Of Shanghai, Chinese Academy Of Sciences Use of phosphorylation pathway-related factor in regulating function of regulatory t cell
JP5668001B2 (en) * 2012-03-05 2015-02-12 丸善製薬株式会社 Skin cosmetics and hair cosmetics
US10279012B2 (en) 2013-03-11 2019-05-07 Healthpartners Research & Education Methods of treating and preventing social communication disorder in patients by intranasal administration of insulin
US10172915B2 (en) * 2013-10-20 2019-01-08 Duke University Methods and compositions for activation of sirtuins with Annexin A1 peptides
US10314911B2 (en) 2014-04-08 2019-06-11 Healthpartners Research & Education Methods for protecting and treating traumatic brain injury, concussion and brain inflammation with intranasal insulin
JP2016172705A (en) * 2015-03-17 2016-09-29 国立大学法人 東京大学 Mint3 inhibitor
WO2017103637A1 (en) 2015-12-18 2017-06-22 Blirt S.A. Diphenylpropane compounds and their cytotoxic activity
JP7029777B2 (en) * 2016-03-28 2022-03-04 国立大学法人金沢大学 A composition for suppressing androgen-dependent or independent prostate cancer cells and a pharmaceutical preparation for prostate cancer containing the same.
CN105949162B (en) * 2016-05-09 2018-02-09 许爱娥 Application of the caffeic acid derivative in leucoderma is treated
US11013673B2 (en) 2016-11-03 2021-05-25 Repairogen Corp. Cosmetic compositions containing quinones and their topical use on skin and hair
EP3534875A1 (en) 2016-11-03 2019-09-11 Repairogen Corp Cosmetic compositions containing quinones and their topical use on skin and hair
WO2019213494A1 (en) 2018-05-03 2019-11-07 Repairogen Corp Cosmetic compositions containing quinones and their topical use on skin and hair
JP2020070242A (en) * 2018-10-29 2020-05-07 アドファーマ株式会社 Hair growth and hair restoration improver
KR102519649B1 (en) * 2019-10-31 2023-04-10 이주운 Composition for the prevention or treatment of prostate-related disease comprising Rhodiola sachalinensis root extract containing kaempferol and epicatechin gallate

Citations (23)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US122777A (en) * 1872-01-16 Improvement in organ actions
US176898A (en) * 1876-05-02 Improvement in compositions for preventing acid fermentation
US5306486A (en) * 1993-03-01 1994-04-26 Elizabeth Arden Co., Division Of Conopco, Inc. Cosmetic sunscreen composition containing green tea and a sunscreen
US5605929A (en) * 1992-05-27 1997-02-25 Arch Development Corp. Methods and compositions for inhibiting 5α-reductase activity
US5665393A (en) * 1996-09-03 1997-09-09 International Medical Research, Inc. Herbal composition for treating prostate carcinoma
US5686082A (en) * 1992-12-24 1997-11-11 L'oreal Cosmetic or pharmaceutical composition containing a combination of a polyphenol and a ginkgo extract
US5788971A (en) * 1993-10-21 1998-08-04 Sky. Food Co., Ltd. Active oxygen free radical scavenging agent
US5804594A (en) * 1997-01-22 1998-09-08 Murad; Howard Pharmaceutical compositions and methods for improving wrinkles and other skin conditions
US5804168A (en) * 1997-01-29 1998-09-08 Murad; Howard Pharmaceutical compositions and methods for protecting and treating sun damaged skin
US5962517A (en) * 1997-01-31 1999-10-05 Murad; Howard Pharmaceutical compositions and methods for treating acne
US6093411A (en) * 1998-03-16 2000-07-25 The Procter & Gamble Company Compositions for regulating skin appearance
US6180662B1 (en) * 1994-12-13 2001-01-30 Beiersdorf Ag Agents acting against hyperreactive and hypoactive, deficient skin conditions and manifest dermatitides
US6183731B1 (en) * 1998-07-07 2001-02-06 Unilever Home & Personal Care Usa Division Of Conopco, Inc. Method of reducing or preventing malodour
US6197808B1 (en) * 1996-11-18 2001-03-06 Cancer Instititute (Hospital), Chinese Academy Of Medical Sciences Methods for treating hyperplasia
US6231877B1 (en) * 1998-07-03 2001-05-15 Lanatech Laboratoire Nature Et Technique Cosmetic composition exploiting synergistic radical scavenging effects
US6248341B1 (en) * 2000-01-14 2001-06-19 Color Access, Inc. Method of treating topical angiogenesis-related disorders
US6337320B1 (en) * 1996-10-11 2002-01-08 Thione International, Inc. Reparatives for ultraviolet radiation skin damage
US6391308B1 (en) * 1996-03-13 2002-05-21 Archer Daniels Midland Company Method of preparing and using isoflavones for the treatment of blood related illnesses
US6444234B1 (en) * 1998-07-07 2002-09-03 Kenneth B Kirby Compositions for rapid and non-irritating transdermal delivery of pharmaceutically active agents and methods for formulating such compositions and delivery thereof
US20020147353A1 (en) * 1999-09-23 2002-10-10 Vereniging Christelijk Wetenschappelijk Anderwijs Novel flavonoids
US6471973B2 (en) * 1998-05-15 2002-10-29 Coletica Flavonoide esters and their use notably in cosmetics
US20020176898A1 (en) * 1999-03-30 2002-11-28 Purdue Research Tea catechins in sustained release formulations as cancer specific proliferation inhibitors
US6696484B2 (en) * 1997-10-31 2004-02-24 University Of Chicago Office Of Technology And Intellectual Property Method and compositions for regulation of 5-alpha reductase activity

Family Cites Families (34)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE2807407A1 (en) 1978-02-17 1979-08-30 Schering Ag TESTOSTERONEESTER
JPS57118580A (en) 1981-01-16 1982-07-23 Kanebo Ltd Novel catechin derivative
EP0116439A3 (en) 1983-02-02 1986-01-29 Suntory Limited Fatty acid containing hair tonic composition
DE3514724A1 (en) * 1985-04-24 1986-10-30 Albin F. Dr. 4200 Oberhausen Jereb Ointment for the prevention and treatment of disorders of the eyes and skin
JPS6284021A (en) * 1985-10-08 1987-04-17 Shiseido Co Ltd Testosterone-5alpha-reductase inhibitor
JPS6425726A (en) 1987-04-24 1989-01-27 Yuutoku Yakuhin Kogyo Kk Agent for eliminating active oxygen free radical
EP0309086A1 (en) 1987-09-07 1989-03-29 Efamol Holdings Plc Treatment of male pattern baldness and of unwanted hair growth
JPH0196124A (en) * 1987-10-07 1989-04-14 Fujirebio Inc Carcinostatic agent
JP2753607B2 (en) * 1988-08-11 1998-05-20 一丸ファルコス株式会社 Testosterone 5α-reductase inhibitor
JPH02300120A (en) 1989-05-11 1990-12-12 Nippon Oil & Fats Co Ltd Carcinostatic agent
JPH03141220A (en) * 1989-10-26 1991-06-17 Tsumura & Co Antiretroviral agent
JPH07112979B2 (en) 1991-11-13 1995-12-06 株式会社スカイ・フード Active oxygen free radical scavenger
JP3211191B2 (en) 1992-03-31 2001-09-25 太陽化学株式会社 Tea extract composition
US5422371A (en) 1992-05-27 1995-06-06 Arch Development Corp. Methods and compositions for inhibiting 5α-reductase activity
JPH06128168A (en) 1992-10-14 1994-05-10 Suetsuna Yoko Tea catechin compounds having antimutagenic activity and superoxide dismutage-like activity
JPH07112980B2 (en) 1992-11-04 1995-12-06 株式会社スカイ・フード Active oxygen free radical scavenger
CN1079361A (en) 1993-04-24 1993-12-15 河南中医学院 Instant green tea and processing technology thereof
FR2711060B1 (en) * 1993-10-13 1995-11-17 Oreal Method for modifying the growth of hair and / or hair and compositions which can be used for this purpose.
IL112205A0 (en) * 1994-01-06 1995-03-15 Res Dev Foundation Curcumin, analogues of curcumin and novel uses thereof
FR2734478B1 (en) 1995-05-23 1997-07-11 Berkem Sa COSMETIC OR PHARMACEUTICAL COMPOSITION FOR TOPICAL OR ORAL USE HAVING ANTIRADICAL PROPERTIES AND A FILTERING POWER AGAINST UV
JPH08337510A (en) * 1995-06-15 1996-12-24 Advanced Sukin Res Kenkyusho:Kk Melanin formation inhibitor
JPH09176011A (en) * 1995-12-27 1997-07-08 Kureha Chem Ind Co Ltd Flavonoid-containing agent for suppressing synthesis of protein of hsp27 family
WO1997027848A1 (en) * 1996-01-31 1997-08-07 Board Of Regents, The University Of Texas System SENSITIZATION OF HER2/neu OVER-EXPRESSING CANCER CELLS TO CHEMOTHERAPEUTIC DRUGS
JPH09315985A (en) 1996-05-29 1997-12-09 Kanebo Ltd Suppressant for mutation
DE19627344A1 (en) 1996-07-01 1998-01-08 Vitasyn Gmbh Entwicklung & Ver Therapeutic composition containing epicatechin and/or theaflavin
US5665367A (en) 1996-09-27 1997-09-09 Chesebrough-Pond's Usa Co., Division Of Conopco, Inc. Skin care compositions containing naringenin and/or quercetin and a retinoid
WO1998048790A1 (en) * 1997-04-28 1998-11-05 Anticancer, Inc. Use of genistein and related compounds to treat certain sex hormone related conditions
JP4738592B2 (en) 1997-10-31 2011-08-03 アーチ・デヴェロップメント・コーポレイション Methods and compositions for modulating 5α-reductase activity
DE29813269U1 (en) 1998-07-25 1998-11-05 Schmitt Angelo tablet
US6428818B1 (en) 1999-03-30 2002-08-06 Purdue Research Foundation Tea catechin formulations and processes for making same
US6156912A (en) 1999-04-09 2000-12-05 Mars, Incorporated 88, 66, and 68 catechin and epicatechin dimers and methods for their preparation
WO2001026668A1 (en) 1999-10-14 2001-04-19 Unilever N.V. Compositions with anti-prostate cancer activity
FI20000004A0 (en) 2000-01-03 2000-01-03 Slk Foundation Flavonoidilääke ...
ES2225123T3 (en) 2000-04-04 2005-03-16 Color Access, Inc. COMPOSITION TO IMPROVE THE FUNCTION OF THE SKIN LIPIDIC BARRIER.

Patent Citations (24)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US176898A (en) * 1876-05-02 Improvement in compositions for preventing acid fermentation
US122777A (en) * 1872-01-16 Improvement in organ actions
US5605929A (en) * 1992-05-27 1997-02-25 Arch Development Corp. Methods and compositions for inhibiting 5α-reductase activity
US5686082A (en) * 1992-12-24 1997-11-11 L'oreal Cosmetic or pharmaceutical composition containing a combination of a polyphenol and a ginkgo extract
US5306486A (en) * 1993-03-01 1994-04-26 Elizabeth Arden Co., Division Of Conopco, Inc. Cosmetic sunscreen composition containing green tea and a sunscreen
US5788971A (en) * 1993-10-21 1998-08-04 Sky. Food Co., Ltd. Active oxygen free radical scavenging agent
US6180662B1 (en) * 1994-12-13 2001-01-30 Beiersdorf Ag Agents acting against hyperreactive and hypoactive, deficient skin conditions and manifest dermatitides
US6395279B1 (en) * 1996-03-13 2002-05-28 Archer Daniels Midland Company Method of preparing and using isoflavones for the treatment of cancer
US6391308B1 (en) * 1996-03-13 2002-05-21 Archer Daniels Midland Company Method of preparing and using isoflavones for the treatment of blood related illnesses
US5665393A (en) * 1996-09-03 1997-09-09 International Medical Research, Inc. Herbal composition for treating prostate carcinoma
US6337320B1 (en) * 1996-10-11 2002-01-08 Thione International, Inc. Reparatives for ultraviolet radiation skin damage
US6197808B1 (en) * 1996-11-18 2001-03-06 Cancer Instititute (Hospital), Chinese Academy Of Medical Sciences Methods for treating hyperplasia
US5804594A (en) * 1997-01-22 1998-09-08 Murad; Howard Pharmaceutical compositions and methods for improving wrinkles and other skin conditions
US5804168A (en) * 1997-01-29 1998-09-08 Murad; Howard Pharmaceutical compositions and methods for protecting and treating sun damaged skin
US5962517A (en) * 1997-01-31 1999-10-05 Murad; Howard Pharmaceutical compositions and methods for treating acne
US6696484B2 (en) * 1997-10-31 2004-02-24 University Of Chicago Office Of Technology And Intellectual Property Method and compositions for regulation of 5-alpha reductase activity
US6093411A (en) * 1998-03-16 2000-07-25 The Procter & Gamble Company Compositions for regulating skin appearance
US6471973B2 (en) * 1998-05-15 2002-10-29 Coletica Flavonoide esters and their use notably in cosmetics
US6231877B1 (en) * 1998-07-03 2001-05-15 Lanatech Laboratoire Nature Et Technique Cosmetic composition exploiting synergistic radical scavenging effects
US6183731B1 (en) * 1998-07-07 2001-02-06 Unilever Home & Personal Care Usa Division Of Conopco, Inc. Method of reducing or preventing malodour
US6444234B1 (en) * 1998-07-07 2002-09-03 Kenneth B Kirby Compositions for rapid and non-irritating transdermal delivery of pharmaceutically active agents and methods for formulating such compositions and delivery thereof
US20020176898A1 (en) * 1999-03-30 2002-11-28 Purdue Research Tea catechins in sustained release formulations as cancer specific proliferation inhibitors
US20020147353A1 (en) * 1999-09-23 2002-10-10 Vereniging Christelijk Wetenschappelijk Anderwijs Novel flavonoids
US6248341B1 (en) * 2000-01-14 2001-06-19 Color Access, Inc. Method of treating topical angiogenesis-related disorders

Cited By (25)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20050171084A1 (en) * 2002-03-27 2005-08-04 Cairns William J. Methods of treatment with lxr modulators
US20050175565A1 (en) * 2002-06-21 2005-08-11 L'oreal Use of taurine for the treatment of alopecia
US9138408B2 (en) 2002-06-21 2015-09-22 L'oreal Use of taurine for treating alopecia
US20060159714A1 (en) * 2003-07-07 2006-07-20 Jean-Noel Thorel Use of an antioxidant in a dermatological and/or cosmetic composition
US8586030B2 (en) 2004-01-22 2013-11-19 University Of Miami Co-enzyme Q10 formulations and methods of use
US8147825B2 (en) 2004-01-22 2012-04-03 University Of Miami Topical co-enzyme Q10 formulations and methods of use
US8771680B2 (en) 2004-01-22 2014-07-08 University Of Miami Topical co-enzyme Q10 formulations and methods of use
US8562976B2 (en) 2004-01-22 2013-10-22 University Of Miami Co-enzyme Q10 formulations and methods of use
US20100034904A1 (en) * 2007-03-21 2010-02-11 Giuliani S.P.A. Composition comprisong rutin and polyunsaturated fatty acid having an inhibitory activity on 5-alph reductase
US9144560B2 (en) * 2007-03-21 2015-09-29 Giuliani S.P.A. Composition comprising rutin and polyunsaturated fatty acid having an inhibitory activity on 5 α-reductase
US8454945B2 (en) 2007-03-22 2013-06-04 Berg Pharma Llc Topical formulations having enhanced bioavailability
US10588859B2 (en) 2007-03-22 2020-03-17 Berg Llc Topical formulations having enhanced bioavailability
US8557864B2 (en) 2007-09-19 2013-10-15 Nagoya Industrial Science Research Institute Agent having neurotrophic factor-like activity
US10668028B2 (en) 2008-04-11 2020-06-02 Berg Llc Methods and use of inducing apoptosis in cancer cells
US10351915B2 (en) 2009-05-11 2019-07-16 Berg Llc Methods for treatment of oncological disorders using an epimetabolic shifter (Coenzyme Q10)
US10519504B2 (en) 2009-05-11 2019-12-31 Berg Llc Methods for treatment of oncological disorders using epimetabolic shifters, multidimensional intracellular molecules, or environmental influencers
US9896731B2 (en) 2009-05-11 2018-02-20 Berg Llc Methods for treatment of oncological disorders using an epimetabolic shifter (coenzyme Q10)
US11028446B2 (en) 2009-05-11 2021-06-08 Berg Llc Methods for treatment of oncological disorders using an epimetabolic shifter (coenzyme Q10)
US11400058B2 (en) 2010-03-12 2022-08-02 Berg Llc Intravenous formulations of coenzyme Q10 (CoQ10) and methods of use thereof
US10376477B2 (en) 2011-04-04 2019-08-13 Berg Llc Method of treating or preventing tumors of the central nervous system
US11452699B2 (en) 2011-04-04 2022-09-27 Berg Llc Method of treating or preventing tumors of the central nervous system
US10973763B2 (en) 2011-06-17 2021-04-13 Berg Llc Inhalable pharmaceutical compositions
US10933032B2 (en) 2013-04-08 2021-03-02 Berg Llc Methods for the treatment of cancer using coenzyme Q10 combination therapies
US9901542B2 (en) 2013-09-04 2018-02-27 Berg Llc Methods of treatment of cancer by continuous infusion of coenzyme Q10
US11298313B2 (en) 2013-09-04 2022-04-12 Berg Llc Methods of treatment of cancer by continuous infusion of coenzyme Q10

Also Published As

Publication number Publication date
WO1999022728A1 (en) 1999-05-14
JP4738592B2 (en) 2011-08-03
EP1027045A1 (en) 2000-08-16
EP1027045A4 (en) 2004-12-08
KR20010031501A (en) 2001-04-16
JP2003524577A (en) 2003-08-19
US6576660B1 (en) 2003-06-10
AU1289899A (en) 1999-05-24

Similar Documents

Publication Publication Date Title
US6576660B1 (en) Methods and compositions for regulation of 5-α-reductase activity
US5605929A (en) Methods and compositions for inhibiting 5α-reductase activity
US6696484B2 (en) Method and compositions for regulation of 5-alpha reductase activity
US5422371A (en) Methods and compositions for inhibiting 5α-reductase activity
JP2002541158A (en) Hydroxymatairesinol in cancer prevention
EP1927352A2 (en) Therapeutic methods and compositions involving isoflavones
US20120156150A1 (en) Composition for Improving Skin Condition Comprising Extract from Black Tea as Active Ingredient
Liao et al. Growth suppression of hamster flank organs by topical application of catechins, alizarin, curcumin, and myristoleic acid
US6878381B2 (en) Resorcinol composition
KR101256588B1 (en) Composition including beta-carboline alkaloid for stimulating melanin production
Mccoy et al. Botanical extracts for the treatment of androgenetic alopecia
BG65146B1 (en) Pharmaceutical composition for external application and use thereof as a skin lightening agent or for reducing of human pigmentation
CA3095167A1 (en) Skin care compositions and their applications
JP4754178B2 (en) Promotion of melanin production by lignans
JP4694068B2 (en) Testosterone-5α-reductase inhibitor
JP2020502050A (en) Skin protection composition containing dendrobium-based material
EP0621030B1 (en) Use of benzomorpholine derivatives in skin whitening cosmetic or dermopharmaceutical compositions
EP0623340B1 (en) Use of hydroxy or dihydroxynaphthalene in the preparation of a cosmetic or dermatological composition for skin whitening
JP4279523B2 (en) Tyrosinase inhibitor
CN115836979A (en) 4-butyl resorcinol liposome and preparation method thereof
Liang et al. Growth Suppression of Hamster Flank Organs by Topical Application of 7-Linolenic and Other Fatty Acid Inhibitors of 5a-Reductase
Voegeli et al. 5a-Reductase and Its Inhibitors
TW201632185A (en) Skin whitening composition prepared with imatininb and application thereof

Legal Events

Date Code Title Description
AS Assignment

Owner name: UNIVERSITY OF CHICAGO OFFICE OF TECHNOLOGY AND INT

Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:LIAO, SHUTSUNG;HIIPAKKA, ROBERT;REEL/FRAME:013636/0531

Effective date: 20021122

STCB Information on status: application discontinuation

Free format text: ABANDONED -- FAILURE TO RESPOND TO AN OFFICE ACTION